Effect of short-term geo-stimulation on the distribution of32P in decapitated pea seedlings

Three-leaf pea (Pisum sativum L. cv. ‘Alaska’) seedlings were oriented horizontally and³²P was applied to leaf-2 for 6 h period, at the end of which the distribution of isotopes in the seedlings was determined. It was found that (i) isotope accumulation in the apices of the vertical and horizontal seedlings remained almost the same; (ii) on decapitation almost all the isotopes in the apex diverted to the roots in the vertical seedlings; (iii) among the horizontal seedlings³²P was retained in the hanging treated leaf and the rest moved to the roots via shoots; (iv) the dry weight of the hanging treated leaf was slightly greater than the above-axis one; (v) the position of buds on the axis had no effect on the initial³²P accumulation in them, however, a trend of greater isotope accumulation was noted in the above-axis basal buds; (vi) geo.stimulation had no effect on the total dry weight of the seedlings; and (vii) in geo-stimulated seedlings decrease in the dry weights of shoots and its concomittant increase in roots was primarily the result of the movement of substances.     

Energy cost and metabolic regulation during intermittent and continuous tetanic contractions in human skeletal muscle

Muscle ATP turnover, glycogenolytic, and glycolytic rates were estimated to compare the energy cost and glycolytic regulation of 102.4 s of continuous and intermittent stimulation. Quadriceps femoris muscles of male subjects were stimulated at 20 Hz for one continuous contraction (n = 6) or a series of 64 contractions (1.6 s on, 1.6 s off; n = 6). Leg blood flow was occluded and muscle biopsies were obtained at rest and following 51.2 and 102.4 s of contraction time in both conditions. Isometric force production by the activated knee extensors decreased to 55% of initial contraction force with intermittent and 80% of initial contraction force with continuous stimulation following 51.2 s of contraction time. Corresponding ATP turnover rates were 4.49 +/- 0.39 and 3.80 +/- 0.44 mmol.kg dry muscle-1.s-1. When normalized for tension production the respective energy costs of intermittent and continuous contractions were 3.66 +/- 0.47 and 2.64 +/- 0.36 mmol ATP.kg-1.100 N-1. Glycogenolytic rates were identical during the first 51.2 s of stimulation but glycolysis was higher in the intermittent group (1.05 +/- 0.10 vs. 0.86 +/- 0.11 mmol.kg-1.s-1). We suggest that the increased ATP utilization of intermittent contractions is associated with enhanced Ca2+-transport ATPase activity during relaxation and enhanced actomyosin ATPase activity during the early portion of each contraction. Glycolytic rate is dependent on ATP demand and regulated by allosteric modulators of phosphofructokinase and pyruvate kinase which are released or consumed in the reactions associated with contraction.     

NCS-1 Expression in Rat Brain after Electroconvulsive Stimulation

Although electroconvulsive therapy (ECT) has been used as a treatment for mental disorder since 1930s, little progress has been made towards understanding the mechanisms underlying its therapeutic and adverse effects. The aim of this work was to analyze the expression of NCS-1 (neuronal calcium sensor 1, a protein that was found to be altered in post-mortem prefrontal cortex of schizophrenic patients) in striatum, cortex, hippocampus and cerebellum of Wistar rats after acute or chronic electroconvulsive stimulation (ECS). Rats were submitted to a single stimulation (acute) or to a series of eight stimulations, applied one every 48 h (chronic). Animals were killed for collection of tissue samples at time zero, 30 min, 3, 12, 24 and 48 h after stimulation in the acute model and at the same time intervals after the last stimulation in the chronic model. Our results indicated that chronic ECS increased the expression of NCS-1 only in cerebellum. Such results on the expression of proteins involved in signaling pathways that are relevant for neuropsychiatric disorders and treatment, in particular ECT, can contribute to shed light on the mechanisms related to therapeutic and adverse effects.     

Modification of a three-compartment muscle fatigue model to predict peak torque decline during intermittent tasks

This study aimed to test whether adding a rest recovery parameter, r, to the analytical three-compartment controller (3CC) fatigue model (Xia and Frey Law, 2008) will improve fatigue estimates during intermittent contractions. The 3CC muscle fatigue model uses differential equations to predict the flow of muscle between three muscle states: Resting (M_R), Active (M_A), and Fatigued (M_F). This model uses a feedback controller to match the active state to target loads and two joint-specific parameters: F, fatigue rate controlling flow from active to fatigued compartments) and R, the recovery rate controlling flow from the fatigued to the resting compartments. This model does well to predict intensity-endurance time curves for sustained isometric tasks. However, previous studies find when rest intervals are present that the model over predicts fatigue. Intermittent rest periods would allow for the occurrence of subsequent reactive vasodilation and post-contraction hyperemia. We hypothesize a modified 3CC-r fatigue model will improve predictions of force decay during intermittent contractions with the addition of a rest recovery parameter, r, to augment recovery during rest intervals, representing muscle re-perfusion. A meta-analysis compiling intermittent fatigue data from 63 publications reporting decline in peak torque (% torque decline) were used for comparison. The original model over-predicted fatigue development from 19 to 29% torque decline; the addition of a rest multiplier significantly improved fatigue estimates to 6–10% torque decline. We conclude the addition of a rest multiplier to the three-compartment controller fatigue model provides a physiologically consistent modification for tasks involving rest intervals, resulting in improved estimates of muscle fatigue.     

Statistical analysis of mediator release from the cyto-neural junction of the posterior canal of the labyrinth isolated from the frog]

The transmitter release mechanism was investigated at the cyto-neural junction of the frog labyrinth posterior canal. Low frequency (less than 100/s) non overlapping EPSPs were intracellularly recorded both at rest and during inhibitory mechanical stimulation of the canal (2-8 deg/s2). Recordings were obtained: in control solution; in the presence of increased external Ca2+ (9 mM); in Ca-free EGTA (5 mM) solution and during electrical activation at 50 Hz of the posterior canal inhibitory efferent system. Individual synaptic potentials were digitized and their peak amplitudes, their time integrals as well as the time intervals between them were evaluated. The time intervals proved to be exponentially distributed, suggesting a random EPSP occurrence. The analytical reconstruction of the EPSP waveform indicated that a gamma- function fitted reasonably well both the single and averaged events. As regards the averaged event, despite the scatter in the values of the gamma-function exponential factor (range 1.1-2.2), in the EPSP time-to-peak (0.6-1.2 ms) and peak amplitude (0.9-2.7 mV) displayed by the units, no significant differences were observed in the same fibre between control and test conditions. Moreover, the event peak amplitude distribution represented by cumulative plots or amplitude histograms was fitted by a lognormal function. The distributions obtained for the same unit in control solution proved to be not significantly different from those successively obtained under test conditions. The unimodal and continuous EPSP distributions, together with the unvarying characteristics of the single events, strongly suggest that the observed potentials are true mEPSPs due to the release of single quanta of transmitter.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hydrotropism and Its Interaction with Gravitropism in Maize Roots

We have partially characterized root hydrotropism and its interaction with gravitropism in maize (Zea mays L.). Roots of Golden Cross Bantam 70, which require light for orthogravitropism, showed positive hydrotropism; bending upward when placed horizontally below a hydrostimulant (moist cheesecloth) in 85% relative humidity (RH) and in total darkness. However, the light-exposed roots of Golden Cross Bantam 70 or roots of a normal maize cultivar, Burpee Snow Cross, showed positive gravitropism under the same conditions; bending downward when placed horizontally below the hydrostimulant in 85% RH. Light-exposed roots of Golden Cross Bantam 70 placed at 70° below the horizontal plane responded positively hydrotropically, but gravitropism overcame the hydrotropism when the roots were placed at 45° below the horizontal. Roots placed vertically with the tip down in 85% RH bent to the side toward the hydrostimulant in both cultivars, and light conditions did not affect the response. Such vertical roots did not respond when the humidity was maintained near saturation. These results suggest that hydrotropic and gravitropic responses interact with one another depending on the intensity of one or both factors. Removal of the approximately 1.5 millimeter root tip blocked both hydrotropic and gravitropic responses in the two cultivars. However, removal of visible root tip mucilage did not affect hydrotropism or gravitropism in either cultivar.     

Selected contribution: acute cellular and molecular responses to resistance exercise.

Training protocols apply sequential bouts of resistance exercise (RE) to induce the cellular and molecular responses necessary to produce compensatory hypertrophy. This study was designed to 1) define the time course of selected cellular and molecular responses to a single bout of RE and 2) examine the effects of interbout rest intervals on the summation of these responses. Rat muscles were exposed to RE via stimulation of the sciatic nerve in vivo. Stimulated and control muscles were obtained at various time points post-RE and analyzed via Western blot and RT-PCR. A single bout of RE increased intracellular signaling (i.e., phosphorylations) and expression of mRNAs for insulin-like growth factor-I system components and myogenic markers (e.g., cyclin D1, myogenin). A rest interval of 48 h between RE bouts resulted in much greater summation of myogenic responses than 24- or 8-h rest intervals. This experimental approach should be useful for studying the regulatory mechanisms that control the hypertrophy response. These methods could also be used to compare and contrast different exercise parameters (e.g., concentric vs. eccentric, etc.).     

Amyloplasts are necessary for full gravitropic sensitivity in roots of Arabidopsis thaliana

The observation that a starchless mutant (TC7) of Arabidopsis thaliana (L.) Heynh. is gravitropic (T. Caspar and B.G. Pickard, 1989, Planta 177, 185–197) raises questions about the hypothesis that starch and amyloplasts play a role in gravity perception. We compared the kinetics of gravitropism in this starchless mutant and the wild-type (WT). Wild-type roots are more responsive to gravity than TC7 roots as judged by several parameters: (1) Vertically grown TC7 roots were not as oriented with respect to the gravity vector as WT roots. (2) In the time course of curvature after gravistimulation, curvature in TC7 roots was delayed and reduced compared to WT roots. (3) TC7 roots curved less than WT roots following a single, short (induction) period of gravistimulation, and WT, but not TC7, roots curved in response to a 1-min period of horizontal exposure. (4) Wild-type roots curved much more than TC7 roots in response to intermittent stimulation (repeated short periods of horizontal exposure); WT roots curved in response to 10 s of stimulation or less, but TC7 roots required 2 min of stimulation to produce a curvature. The growth rates were equal for both genotypes. We conclude that WT roots are more sensitive to gravity than TC7 roots. Starch is not required for gravity perception in TC7 roots, but is necessary for full sensitivity; thus it is likely that amyloplasts function as statoliths in WT Arabidopsis roots. Furthermore, since centrifugation studies using low gravitational forces indicated that starchless plastids are relatively dense and are the most movable component in TC7 columella cells, the starchless plastids may also function as statoliths.Abbreviations S2 story two - S3 story three - WT wild-type     

Manipulating vibratory devices’ orientation and position enhances proprioceptive disturbance during upright stance

We tested local vibration effects during upright standing considering: (i) the orientation of vibratory devices in relation to muscle fibres; (ii) the muscle region stimulated; and (iii) the number of stimulation spots. Results showed a higher balance disturbance with vibration devices oriented parallel to triceps surae muscle fibres. The single stimulation of the proximal region of the tibialis anterior muscle belly induces the same proprioceptive disturbance as stimulating multiple regions simultaneously.     

The effects of interval training on oxygen pulse and performance in supra-threshold runs

The aim of this study was to examine (i) the effects of a severe interval training period on oxygen pulse kinetics (O2-p, the ratio between VO2 and heart rate), and (ii) to study the consequences of these effects on the variation of performance (time to exhaustion) during severe runs. Seven athletes were tested before and after an eight-weeks period of a specific intermittent training at v Delta 50, i.e., the intermediate velocity between the lactate threshold (vLT) and the velocity associated with VO2max (vVO2max ). During the test sessions, athletes performed an incremental test and an all-out test at the pretraining v Delta 50. After the training period they also completed an additional all-out test at the posttraining v Delta 50 (v Delta 50bis). Results showed that after training there was i) an increase in the O2-p maximal value during the incremental test (22.7 +/- 1.5 mlO2.b-1 vs. 20.6 +/- 1.5 mlO2.b-1; p < 0.04), ii) a decrease in the time to reach the O2-p steady state (TRO2-p ) at the same absolute v Delta 50 (33 +/- 7 s vs. 60 +/- 27 s; p < 0.04) and iii) an increase in the O2-p steady state duration (TSSO2-p) at the same absolute v Delta 50 (552 +/- 201 s vs. 407 +/- 106 s; p < 0.04). However, there was no relationship between the improvement of these two O 2 -p kinetics parameters (TRO2-p and TSS O2-p) and those of the performance. This study found that after an individualised interval-training program conducted at the same absolute velocity, the O2-p kinetics reached a steady state quicker and for a longer duration than before training. This is however not related with the improvement of performance.     

Intermittent Pool Beds Are Permanent Cyclic Habitats with Distinct Wet,Moist and Dry Phases

Recognition that intermittent pools are a single habitat phase of an intermittent pool bed that cycles between aquatic and terrestrial habitat greatly enhances their usefulness for addressing general questions in ecology. The aquatic phase has served as a model system in many ecological studies, because it has distinct habitat boundaries in space and time and is an excellent experimental system, but the aquatic to terrestrial transition and terrestrial phase remain largely unstudied. We conducted a field experiment within six replicate natural intermittent pool beds to explore macroinvertebrate community dynamics during the transition from aquatic to terrestrial habitat and during the terrestrial phase. We monitored and compared macroinvertebrate communities within leaf packs that i) remained wet, ii) underwent drying (i.e., started wet and then dried), and iii) remained dry. Our results show that i) a diverse macroinvertebrate community inhabits all phases of intermittent pool beds, ii) pool drying involves colonization by an assemblage of macroinvertebrates not recorded in permanently terrestrial leaf packs, iii) the community within dried leaf packs remains distinct from that of permanently terrestrial leaf packs for an extended period following drying (possibly until subsequent refilling), and iv) there are likely to be strong spatial and temporal resource linkages between the aquatic and terrestrial communities. The unique environmental characteristics of intermittent pool beds, which repeatedly cycle from aquatic to terrestrial habitat, should continue to make them valuable study systems.     

Phototropic Responses of Vaucheria geminata to Intermittent Blue Light Stimuli

Phototropic responses of a tip-growing coenocytic alga Vaucheria geminata to intermittent blue light pulses were analyzed. Curvatures caused by repeated light pulses separated by dark intervals of various lengths were as large as those obtained by continuous light of the same incident energy, unless the length of the dark interval exceeded a critical value of about 30 to 40 seconds. If the dark interval was longer than the critical length, bending no longer took place. Another response to intermittent irradiation was found. In a narrow range of light pulse and dark interval lengths, i.e. light pulses longer than 10 milliseconds and dark intervals shorter than a value ranging from 15 to 150 milliseconds, intermittent irradiation was more effective than continuous irradiation of the same total energy. In this region, the maximum response to repeated pulses was equal to the curvature caused by a continuous irradiation of the same total elapsed time. It is evident that in this condition the dark interval is not perceived and the alga responds as if it were in light. The results suggest that two different dark reactions may be involved in the phototropic response of this algal cell.     

Acid-Efflux Patterns of Primary and Secondary Roots of Ricinus communis (Euphorbiaceae)

Graviresponding primary roots of castor bean (Ricinus communis)were characterized by more acid efflux on their upper (i.e.rapidly growing) sides than their lower sides. Acid-efflux patternsof the upper and lower sides of horizontally oriented lateralroots were symmetrical. The onset of graviresponsiveness bysecondary roots correlated positively with the development ofasymmetric patterns of acid efflux similar to those of gravirespondingprimary roots. The addition of 1 mM sodium orthovanadate (aninhibitor of auxin-induced proton efflux) to the growth mediumabolished gravicurvature and the development of acid-effluxasymmetry by primary and secondary roots. These results suggestthat (i) the absence of an auxin-induced asymmetry of acid effluxmay be the factor responsible for uncoupling graviperceptionfrom gravitropism in lateral roots, and (ii) the developmentof an auxin-induced asymmetry of acid efflux may mediate theonset of graviresponsiveness by secondary roots of R. communis. Ricinus communis, castor bean, acid efflux, root gravitropism     

Synaptic vesicle pools at the frog neuromuscular junction

We have characterized the morphological and functional properties of the readily releasable pool (RRP) and the reserve pool of synaptic vesicles in frog motor nerve terminals using fluorescence microscopy, electron microscopy, and electrophysiology. At rest, about 20% of vesicles reside in the RRP, which is depleted in about 10 s by high-frequency nerve stimulation (30 Hz); the RRP refills in about 1 min, and surprisingly, refilling occurs almost entirely by recycling, not mobilization from the reserve pool. The reserve pool is depleted during 30 Hz stimulation with a time constant of about 40 s, and it refills slowly (half-time about 8 min) as nascent vesicles bud from randomly distributed cisternae and surface membrane infoldings and enter vesicle clusters spaced at regular intervals along the terminal. Transmitter output during low-frequency stimulation (2-5 Hz) is maintained entirely by RRP recycling; few if any vesicles are mobilized from the reserve pool.     

Modification of the gravitropic response of seedling roots of raneseed (Brassica napus) transformed by Agrobacterium rhizogenes A4

We have examined the growth and gravitropic response of seedling roots of rapeseed ( Brassica napus . CrGC5–1) transformed by Agrobacterium rhizogenes A4, in order to evaluate if this could constitute a new model system for the study of gravitropism. The transformed clone chosen for study had integrated full-length TL- and TR-DNA from pRi (the root inducing plasmid), and thus included all of the agrobacterial genes potentially involved in the modified phenotype of transformed plants. In the vertical position, the growth rate of transformed roots was higher than controls. During 24 h of continuous stimulation, the optimal angle for gravitropic bending in normal roots was 135° (with respect to the gravity axis), with decreasing response at 90° and 45°. For transformed roots, slight curvature developed at 45° and at 90°, and stronger curvature was observed at 135°, though transformed roots tips never reached the vertical position. The minimum stimulation time necessary to elicit a response (presentation time) was also determined: it was signficantly shorter in normal roots (80 s) than in transformed ones (120 s). The results show that pRi transformed roots are less sensitive to gravity than normal roots.     

Myoelectrical and mechanical changes linked to length specificity during isometric training

Force generation and ATP utilization under anaerobic conditions were studied in the quadriceps femoris muscle of six volunteers. Electrical stimulation (20 Hz) was used to produce contractions with a duration of 0.8 s in one leg and contractions with a duration of 3.2 s in the other leg. The two procedures were designed to give the same total contraction time of 51 s and used the same number of stimulation pulses. Muscle biopsies were taken at rest and after 22 and 51 s of work and analyzed for ATP, phosphocreatine, and glucolytic intermediates. The results were compared with previous studies on continuous and intermittent stimulation. Fatigue developed significantly faster with contractions of short duration, and the energy cost was higher. Since force at the end of stimulation had a negative correlation to ATP utilization, there is no indication that the energy resources limit force generation. By comparison of stimulations producing the same amount of isometric work but with a different number of contractions, we estimate that the energy cost for activation and relaxation of a 1-s contraction is approximately 37% of the total ATP consumption.     

Dissociation induced by voluntary movement between two different components of the centro-parietal P40 SEP to tibial nerve stimulation

Whether the two earliest cortical somatosensory evoked potentials (SEPs) to tibial nerve stimulation (N37 and P40) are generated by the same dipolar source or, instead, originate from different neuronal populations is still a debated problem. We recorded the early scalp SEPs to tibial nerve stimulation in 10 healthy subjects at rest and during voluntary movement of the stimulated foot. We found that the P40, which reached its highest amplitude on the vertex at rest, changed its topography during movement, since its amplitude was reduced much more in the central than in the parietal traces. These findings suggest that two different components contribute to the centro-parietal positivity at rest: (1) the P37 response, which is parietally distributed and is not modified by movement, and (2) the `real' P40 SEP, which is focused on the vertex and is reduced in amplitude during voluntary movement. Since, also, the N37 response did not vary its amplitude under interference condition, it is possible that the N37 and P37 potentials are generated by the same dipolar source. Other later components, namely P50 and N50, were significantly reduced in amplitude during foot movement. Lastly, the subcortical P30 far-field remained unchanged and this suggests that the phenomenon of amplitude reduction during movement (i.e. gating) occurs above the cervico-medullary junction.     

The polarity of statocytes and the gravisensitivity of roots are dependent on the concentration of calcium in statocytes

In order to examine a possible role of calcium in graviperception, the calcium ionophore A23187 was used to elevate the concentration of free cytoplasmic calcium in statocytes of the roots of Lepidium sativum L. After a brief incubation (30 min) in a medium that contained 10 micromoles A23187 and 5.5 micromoles CaCl2, 50% of the roots bent gravitropically during a subsequent 2 h of horizontal exposure, with an angle of curvature that varied from 5 degrees to 70 degrees. The corresponding statocytes exhibited a polar arrangement of cell organelles as did the controls. However, in statocytes from 50% of the roots which were not curved after gravistimulation a portion of the distal endoplasmic reticulum (ER) complex was displaced in the direction of gravity within 30 min of horizontal exposure. After washing of the briefly treated roots for 24 h with 1% dimethylsulfoxide the percentage of gravitropically bending roots increased to approximately 80%, but the angle of curvature amounted to only 5 degrees-10 degrees. Longer treatment (2 h) with A23187 caused a complete loss of graviresponsiveness which was accompanied by disintegration of statocyte polarity. We concluded from these results that i) calcium is involved in graviperception and ii) gravisensitivity depends on the integrity of statocytes.     

Electrical stimulation of human forearm extensor muscles as an indicator of handgrip fatigue and recovery

A set-up for percutaneous electrical stimulation of the forearm extensor muscles and measurement of wrist extension force is described. The frequency-force relationship and pulse duration-force relationship are described together with an experimental protocol showing that brief electrical test stimulations do not produce fatigue. In another set of experiments carried out a few weeks later, the subjects performed handgrip contractions: protocol A at 25% of maximal voluntary contraction (MVC) continuously until exhaustion, protocol B at 25% MVC intermittent (contraction + relaxation = 10 + 2 s) until exhaustion, and protocol C at 25% MVC intermittent until half the time to exhaustion. In all experiments, brief electrical stimulations were used to test the degree of fatigue during and up to 24 h after the experiments. There were marked changes in the force during stimulation at 20 and 100 Hz and these changes did not correlate with the increase in intramuscular temperature. Low frequency fatigue persisted for at least 24 h after protocol A and 1 h after protocols B and C. The significance of this is discussed and it is suggested that low frequency fatigue could be used as a sensitive indicator of muscle dysfunction after low and medium intensity exercise.     

Phagocytic response to fully controlled plural stimulation of antigens on macrophage using on-chip microcultivation system

To understand the control mechanism of innate immune response in macrophages, a series of phagocytic responses to plural stimulation of antigens on identical cells was observed. Two zymosan particles, which were used as antigens, were put on different surfaces of a macrophage using optical tweezers in an on-chip single-cell cultivation system, which maintains isolated conditions of each macrophage during their cultivation. When the two zymosan particles were attached to the macrophage simultaneously, the macrophage responded and phagocytosed both of the antigens simultaneously. In contrast, when the second antigen was attached to the surface after the first phagocytosis had started, the macrophage did not respond to the second stimulation during the first phagocytosis; the second phagocytosis started only after the first process had finished. These results indicate that (i) phagocytosis in a macrophage is not an independent process when there are plural stimulations; (ii) the response of the macrophage to the second stimulation is related to the time" delay from the first stimulation. Stimulations that occur at short time intervals resulted in simultaneous phagocytosis, while a second stimulation that is delayed long enough might be neglected until the completion of the first phagocytic process.