Changes in gravity affect neuromuscular control, biomechanics, and muscle-tendon mechanics in energy storage and dissipation tasks.

This study evaluates neuromechanical control and muscle-tendon interaction during energy storage and dissipation tasks in hypergravity. During parabolic flights, while 17 subjects performed drop jumps (DJs) and drop landings (DLs), electromyography (EMG) of the lower limb muscles was combined with in vivo fascicle dynamics of the gastrocnemius medialis, two-dimensional (2D) kinematics, and kinetics to measure and analyze changes in energy management. Comparisons were made between movement modalities executed in hypergravity (1.8 G) and gravity on ground (1 G). In 1.8 G, ankle dorsiflexion, knee joint flexion, and vertical center of mass (COM) displacement are lower in DJs than in DLs; within each movement modality, joint flexion amplitudes and COM displacement demonstrate higher values in 1.8 G than in 1 G. Concomitantly, negative peak ankle joint power, vertical ground reaction forces, and leg stiffness are similar between both movement modalities (1.8 G). In DJs, EMG activity in 1.8 G is lower during the COM deceleration phase than in 1 G, thus impairing quasi-isometric fascicle behavior. In DLs, EMG activity before and during the COM deceleration phase is higher, and fascicles are stretched less in 1.8 G than in 1 G. Compared with the situation in 1 G, highly task-specific neuromuscular activity is diminished in 1.8 G, resulting in fascicle lengthening in both movement modalities. Specifically, in DJs, a high magnitude of neuromuscular activity is impaired, resulting in altered energy storage. In contrast, in DLs, linear stiffening of the system due to higher neuromuscular activity combined with lower fascicle stretch enhances the buffering function of the tendon, and thus the capacity to safely dissipate energy.NEW & NOTEWORTHY For the first time, the neuromechanics of distinct movement modalities that fundamentally differ in their energy management function have been investigated during overload systematically induced by hypergravity. Parabolic flight provides a unique experimental setting that allows near-natural movement execution without the confounding effects typically associated with load variation. Our findings show that gravity-adjusted muscle activities are inversely affected within jumps and landings. Specifically, in 1.8 G, typical task-specific differences in neuromuscular activity are reduced during the center of mass deceleration phase, resulting in fascicle lengthening, which is associated with energy dissipation.

Muscle in Variable Gravity: "I Do Not Know Where I Am, But I Know What to Do".

Purpose: Fascicle and sarcomere lengths are important predictors of muscle mechanical performance. However, their regulation during stretch-shortening cycle (SSC) activities in usual and challenging conditions is poorly understood. In this study, we aimed to investigate muscle fascicle and sarcomere behavior during drop jumps (a common SSC activity) in conditions of variable gravity. Methods: Fifteen volunteers performed repeated drop jumps in 1 g, hypo-gravity (0 to 1 g), and hyper-gravity (1 to 2 g) during a parabolic flight. Gastrocnemius medialis (GM) electromyographic activity and fascicle length (Lf) were measured at drop-off, ground contact (GC), minimum ankle joint angle (MAJ), and push-off. GM sarcomere number was estimated by dividing Lf, measured by ultrasound at rest, by published data on GM sarcomere length, and measured in vivo at the same joint angle. Changes in sarcomere length were estimated by dividing GM Lf in each jump phase by sarcomere number calculated individually. The sarcomere force-generating capacity in each jump phase was estimated from the sarcomere length-tension relationship previously reported in the literature. Results: The results showed that, regardless of the gravity level, GM sarcomeres operated in the ascending portion of their length-tension relationship in all the jump phases. Interestingly, although in hypo-gravity and hyper-gravity during the braking phase (GC-MAJ) GM fascicles and sarcomeres experienced a stretch (as opposed to the quasi-isometric behavior in 1 g), at MAJ they reached similar lengths as in 1 g, allowing sarcomeres to develop about the 70% of their maximum force. Conclusion: The observed fascicle behavior during drop jumping seems useful for anchoring the tendon, enabling storage of elastic energy and its release in the subsequent push-off phase for effectively re-bouncing in all gravity levels, suggesting that an innate neuromuscular wisdom enables to perform SSC movements also in challenging conditions.

The Anticipation of Gravity in Human Ballistic Movement.

Stretch-shortening type actions are characterized by lengthening of the pre-activated muscle-tendon unit (MTU) in the eccentric phase immediately followed by muscle shortening. Under 1 g, pre-activity before and muscle activity after ground contact, scale muscle stiffness, which is crucial for the recoil properties of the MTU in the subsequent push-off. This study aimed to examine the neuro-mechanical coupling of the stretch-shortening cycle in response to gravity levels ranging from 0.1 to 2 g. During parabolic flights, 17 subjects performed drop jumps while electromyography (EMG) of the lower limb muscles was combined with ultrasound images of the gastrocnemius medialis, 2D kinematics and kinetics to depict changes in energy management and performance. Neuro-mechanical coupling in 1 g was characterized by high magnitudes of pre-activity and eccentric muscle activity allowing an isometric muscle behavior during ground contact. EMG during pre-activity and the concentric phase systematically increased from 0.1 to 1 g. Below 1 g the EMG in the eccentric phase was diminished, leading to muscle lengthening and reduced MTU stretches. Kinetic energy at take-off and performance were decreased compared to 1 g. Above 1 g, reduced EMG in the eccentric phase was accompanied by large MTU and muscle stretch, increased joint flexion amplitudes, energy loss and reduced performance. The energy outcome function established by linear mixed model reveals that the central nervous system regulates the extensor muscles phase- and load-specifically. In conclusion, neuro-mechanical coupling appears to be optimized in 1 g. Below 1 g, the energy outcome is compromised by reduced muscle stiffness. Above 1 g, loading progressively induces muscle lengthening, thus facilitating energy dissipation.

Anticipation of drop height affects neuromuscular control and muscle-tendon mechanics.

This study examined the effect of drop height on neuromechanical control of the plantarflexors in drop jumps (DJs) before and during ground contact (GC). The effect of anticipation on muscle mechanical configurations was investigated in 22 subjects in three conditions (20, 30, and 40 cm): (i) known, (ii) unknown, or (iii) cheat falling heights (announced 40 cm, but actual drop height was 20 cm). Electromyographic (EMG) activity of the m. gastrocnemius medialis (GM) and other shank muscles was recorded and analyzed before GC and during GC separately for the short-, medium-, and long-latency responses (SLR, MLR, and LLR). Changes in GM fascicle length (LM ) were determined via B-mode ultrasound, and muscle-tendon unit length (LMTU ) was estimated. Peak force (P < .001), rate of force development (RFD) (P = .001) and GM EMG activity prior to (P = .003) and during GC (P = .007) was reduced in the unknown compared with the known conditions (P < .05). The amount of shortening in LMTU during GC in unknown and cheat was less compared with the known conditions (P = .005; P = .049). Changes in LMTU lengthening negatively correlated with changes in GM activity around SLR and MLR (P = .006; P = .02) in known and unknown conditions. Taken together, it seems that the central nervous system applies a protective strategy in the unknown condition by reducing muscle activity to result in a lower muscular stiffness and increased tendinous lengthening prior to and during GC. This might be a mechanism to absorb greater elastic energy in the tendon and reduce the magnitude and rate of muscle lengthening and subsequent stretch-induced muscle damage.

The relationship between leg stiffness, forces and neural control of the leg musculature during the stretch-shortening cycle is dependent on the anticipation of drop height.

PURPOSE: This study aimed at investigating how prior knowledge of drop heights affects proactive and reactive motor control in drop jumps (DJ). METHODS: In 22 subjects, the effect of knowledge of three different drop heights (20, 30, 40 cm) during DJs was evaluated in seven conditions: three different drop heights were either known, unknown or cheated (announced 40 cm, but actual drop height was 20 cm). Peak ground reaction force (Fmax) to body weight (BW) ratio (Fmax/BW) and electromyographic (EMG) activities of three shank and five thigh muscles were assessed 150 ms before and during ground contact (GC). Ankle, knee and hip joint kinematics were recorded in the sagittal plane. RESULTS: Leg stiffness, proactive and reactive EMG activity of the leg muscles diminished in unknown and cheat conditions for all drop heights (7-33% and 2-26%, respectively). Antagonistic co-activation increased in unknown (3-37%). At touchdown, increased flexion in knee (~ 5.3° ± 1.9°) and hip extension (~ 2° ± 0.6°) were observed in unknown, followed by an increased angular excursion in hip (~ 2.3° ± 0.2°) and knee joints (~ 5.6° ± 0.2°) during GC (p < 0.05). Correlations between changes in activation intensities, joint kinematics, leg stiffness and Fmax/BW (p < 0.05) indicate that anticipation changes the neuromechanical coupling of DJs. No dropouts were recorded. CONCLUSION: These findings underline that anticipation influences timing and adjustment of motor responses. It is argued that proactive and reactive modulations associated with diminished activation intensities in leg extensors are functionally relevant in explaining changes in leg stiffness and subsequent decline in performance.

Stumbling reactions in hypo and hyper gravity - muscle synergies are robust across different perturbations of human stance during parabolic flights.

The control of bipedal stance and the capacity to regain postural equilibrium after its deterioration in variable gravities are crucial prerequisites for manned space missions. With an emphasize on natural orthograde posture, computational techniques synthesize muscle activation patterns of high complexity to a simple synergy organization. We used nonnegative matrix factorization to identify muscle synergies during postural recovery responses in human and to examine the functional significance of such synergies for hyper-gravity (1.75 g) and hypo-gravity (0.25 g). Electromyographic data were recorded from leg, trunk and arm muscles of five human exposed to five modes of anterior and posterior support surface translations during parabolic flights including transitional g-levels of 0.25, 1 and 1.75 g. Results showed that in 1 g four synergies accounted for 99% of the automatic postural response across all muscles and perturbation directions. Each synergy in 1 g was correlated to the corresponding one in 0.25 and 1.75 g. This study therefore emphasizes the similarity of the synergy organization of postural recovery responses in Earth, hypo- and hyper-gravity conditions, indicating that the muscle synergies and segmental strategies acquired under terrestrial habits are robust and persistent across variable and acute changes in gravity levels.

Stumbling Reactions in Partial Gravity - Neuromechanics of Compensatory Postural Responses and Inter-Limb Coordination During Perturbation of Human Stance.

Spontaneous changes in gravity play a significant role in interplanetary space missions. To preserve the astronauts' capability to execute mission-critical tasks and reduce the risk of injury in transit and on planetary surfaces, a comprehensive understanding of the neuromuscular control of postural responses after balance deterioration in hypo- or hyper-gravity conditions is essential. Therefore, this study aimed to evaluate the effect of acute gravitational variation on postural adjustments in response to perturbations. Gravitational changes were induced using parabolic flight. Postural set was manipulated by randomly providing unilateral left, bilateral or split perturbations which require balance corrections to restore postural stability. In six subjects, postural reactions were recorded after anterior and posterior surface perturbations for progressively increased gravitational conditions spanning from 0.25 to 1.75 g. Ankle and knee joint kinematics and electromyograms (EMG) of eight leg muscles were recorded prior (PRE) and after perturbation onset. Muscle activation onset latencies and amplitudes in the short-, medium-, and long-latency responses (SLR, MLR, LLR) were assessed. Results demonstrate an increased muscle activity (p < 0.05) and co-contraction in the lower extremities (p < 0.05) prior to perturbation in hypo- and hyper-gravity. After perturbation, reduced muscle onset latencies (p < 0.05) and increased muscle activations in the MLR and LLR (p < 0.05), concomitant with an increased co-contraction in the SLR, were manifested with a progressive rise in gravity. Ankle and knee joint deflections remained unaffected, whereas angular velocities increased (p < 0.05) with increasing gravitation. Effects were more pronounced in bi- compared to unilateral or split perturbations (p < 0.05). Neuro-mechanical adaptations to gravity were more distinct and muscle onset latencies were shorter in the displaced compared to the non-displaced leg. In conclusion, the timing and magnitude of postural reflexes involved in stabilization of bipedal stance are gravity-dependent. The approximately linear relationship between gravity and impulse-directed EMG amplitudes or muscle onset latencies after perturbation indicates that the central nervous system correctly predicts the level of gravity. Moreover, it accurately governs contractions in the antigravity musculature to counterbalance the gravitational pull and to regain upright posture after its disturbance. Importantly, unilateral perturbations evoked fast reflex responses in the synergistic muscles of the non-displaced contralateral leg suggesting a synchronized inter-limb coordination mediated by spinal circuitries.

Anticipation modulates neuromechanics of drop jumps in known or unknown ground stiffness.

With an emphasis on ballistic movements, an accurately anticipated neural control is an essential prerequisite to deliver a motor response coincidentally with the event of ground contact. This study investigated how previous knowledge of the ground condition affects proactive and reactive motor control in drop jumps (DJ). Thereby, human anticipatory capacity of muscle activation was investigated regarding neuromuscular activation, joint kinematics and peak forces associated with DJ performance. In 18 subjects, the effect of knowledge of two different surface conditions during DJs was evaluated. Peak force, ground-contact-time (GCT), rate of force development (RFD) and jump height were assessed. Electromyographic (EMG) activities of the m. soleus (SOL) and gastrocnemius medialis (GM) were assessed for 150ms before (PRE) and during ground contact (GC) for the short-, medium-, and long-latency responses. Ankle and knee joint kinematics were recorded in the sagittal plane.In the unknown conditions peak force, RFD and jump height declined, GCT was prolonged, proactive EMG activity (PRE) in SOL and GM was diminished (P<0.05). During GC, a decline in EMG activity in the unknown condition was manifested for SOL and GM for the SLR, MLR and LLR (P<0.05). Ankle and knee joint deflections during GC were increased in the unknown vs. known condition (P<0.05). Peak force, RFD and jump height were positively correlated to GM-EMG in PRE, SLR, MLR and LLR (P<0.05). Results revealed that proactive and reactive modulations in muscle activity prior and during GC are interrelated to the force-time characteristics and height of the jumps. The unknown condition revealed a comparable neuromuscular activity during pre-activation for both conditions, followed by an inhibition in the subsequent phase after touch down. These findings underline that anticipation is a determining factor influencing timing and adjustment of motor responses to accomplish ballistic movements regarding precision and performance.

The role of KDEL-tailed cysteine endopeptidases of Arabidopsis (AtCEP2 and AtCEP1) in root development.

Plants encode a unique group of papain-type cysteine endopeptidases (CysEP) characterized by a C-terminal KDEL endoplasmic reticulum retention signal (KDEL-CysEP) and an unusually broad substrate specificity. The three Arabidopsis KDEL-CysEPs (AtCEP1, AtCEP2, and AtCEP3) are differentially expressed in vegetative and generative tissues undergoing programmed cell death (PCD). While KDEL-CysEPs have been shown to be implicated in the collapse of tissues during PCD, roles of these peptidases in processes other than PCD are unknown. Using mCherry-AtCEP2 and EGFP-AtCEP1 reporter proteins in wild type versus atcep2 or atcep1 mutant plants, we explored the participation of AtCEP in young root development. Loss of AtCEP2, but not AtCEP1 resulted in shorter primary roots due to a decrease in cell length in the lateral root (LR) cap, and impairs extension of primary root epidermis cells such as trichoblasts in the elongation zone. AtCEP2 was localized to root cap corpses adherent to epidermal cells in the rapid elongation zone. AtCEP1 and AtCEP2 are expressed in root epidermis cells that are separated for LR emergence. Loss of AtCEP1 or AtCEP2 caused delayed emergence of LR primordia. KDEL-CysEPs might be involved in developmental tissue remodeling by supporting cell wall elongation and cell separation.

Stability of Sodium Nitroprusside in 5% Dextrose Stored at 4°C in Polypropylene Syringes Protected from Light.

Sodium nitroprusside is a potent vasodilator employed intraoperatively and within critical care areas. The photolabile pharmaceutical agent has been used for decades and various stability studies have been executed. Due to potential shortages and the desire to batch compound sodium nitroprusside at a concentration of 1 mg/mL in polypropylene syringes, a new stability study was performed. Chromatographic analysis was conducted on a C18 column, with elution via an aqueous phase of 0.01 M sodium phosphate monobasic, adjusted to pH 6.5 with sodium hydroxide, and methanol (97.5:2.5) at a rate of 1 mL/min, and subsequent ultraviolet detection at 210 nm. Triplicate determinations of four samples, stored under refrigeration at 4°C, were obtained initially and on days 2, 5, and 9. Turbidity and pH measurements were performed in conjunction with visual observation on days of chromatographic analysis. Results demonstrate that sodium nitroprusside compounded in 5% dextrose at a concentration of 1 mg/mL, stored at 4°C protected from light in polypropylene syringes, is physically and chemically stable for at least 9 days.

Calculating the refractive index for pediatric parenteral nutrient solutions.

PURPOSE: The utility of refractometric analysis for calculating the refractive index (RI) of compounded parenteral nutrient solutions for pediatric patients was examined. METHODS: An equation for calculating the RI of parenteral nutrient solutions was developed by chemical and linear regression analysis of 154 pediatric parenteral nutrient solutions. This equation was then validated by analyzing 1057 pediatric parenteral nutrition samples. The RI for the parenteral nutrient solutions could be calculated by summing the RI contribution for each ingredient and then adding the RI of water. The RI contribution for each ingredient was determined by multiplying the RI of the manufacturer's concentrate by the volume of the manufacturer's concentrate mixed into the parenteral nutrient solution divided by the total volume of the parenteral nutrient solution. RESULTS: The calculated RI was highly correlated with the measured RI (R(2) = 0.94, p < 0.0001). Using a range of two standard deviations (±0.0045), 99.8% of the samples fell into the comparative range. RIs of electrolytes, vitamins, and trace elements in the concentrations used did not affect the RI, similar to the findings of other studies. CONCLUSION: There was no statistical difference between the calculated RI and the measured RI in the final product of a pediatric parenteral nutrient solution. This method of quality control can be used by personnel compounding parenteral nutrient solutions to confirm the compounding accuracy of dextrose and amino acid concentrations in the final product, and a sample can be sent to the hospital laboratory for electrolyte verification.

KDEL-tailed cysteine endopeptidases involved in programmed cell death, intercalation of new cells, and dismantling of extensin scaffolds.

KDEL-tailed cysteine endopeptidases are a group of papain-type peptidases found in senescing tissue undergoing programmed cell death (PCD). Their genes have so far been cloned and analyzed in 12 angiosperms. They are synthesized as proenzymes with a C-terminal KDEL endoplasmatic reticulum retention signal, which is removed with the prosequence to activate enzyme activity. We previously identified three genes for KDEL-tailed cysteine endopeptidases (AtCEP1, AtCEP2, AtCEP3) in Arabidopsis thaliana. Transgenic plants of A. thaliana expressing ß-glucuronidase (GUS) under the control of the promoters for the three genes were produced and analyzed histochemically. GUS activity was promoter- and tissue-specific GUS activity during seedling, flower, and root development, especially in tissues that collapse during final stages of PCD, and in the course of lateral root formation. KDEL-tailed cysteine endopeptidases are unique in being able to digest the extensins that form the basic scaffold for cell wall formation. The broad substrate specificity is due to the structure of the active site cleft of the KDEL-tailed cysteine endopeptidase that accepts a wide variety of amino acids, including proline and glycosylated hydroxyproline of the hydroxyproline rich glycoproteins of the cell wall.

Dual specificities of the glyoxysomal/peroxisomal processing protease Deg15 in higher plants.

Glyoxysomes are a subclass of peroxisomes involved in lipid mobilization. Two distinct peroxisomal targeting signals (PTSs), the C-terminal PTS1 and the N-terminal PTS2, are defined. Processing of the PTS2 on protein import is conserved in higher eukaryotes. The cleavage site typically contains a Cys at P1 or P2. We purified the glyoxysomal processing protease (GPP) from the fat-storing cotyledons of watermelon (Citrullus vulgaris) by column chromatography, preparative native isoelectric focusing, and 2D PAGE. The GPP appears in two forms, a 72-kDa monomer and a 144-kDa dimer, which are in equilibrium with one another. The equilibrium is shifted on Ca(2+) removal toward the monomer and on Ca(2+) addition toward the dimer. The monomer is a general degrading protease and is activated by denatured proteins. The dimer constitutes the processing protease because the substrate specificity proven for the monomer (Phi-Arg/Lys downward arrow) is different from the processing substrate specificity (Cys-Xxx downward arrow/Xxx-Cys downward arrow) found with the mixture of monomer and dimer. The Arabidopsis genome analysis disclosed three proteases predicted to be in peroxisomes, a Deg-protease, a pitrilysin-like metallopeptidase, and a Lon-protease. Specific antibodies against the peroxisomal Deg-protease from Arabidopsis (Deg15) identify the watermelon GPP as a Deg15. A knockout mutation in the DEG15 gene of Arabidopsis (At1g28320) prevents processing of the glyoxysomal malate dehydrogenase precursor to the mature form. Thus, the GPP/Deg15 belongs to a group of trypsin-like serine proteases with Escherichia coli DegP as a prototype. Nevertheless, the GPP/Deg15 possesses specific characteristics and is therefore a new subgroup within the Deg proteases.

Ricinosomes and endosperm transfer cell structure in programmed cell death of the nucellus during Ricinus seed development.

The ricinosome (precursor protease vesicle) is an organelle found exclusively in plant cells. Ricinosomes contain a 45-kDa pro-cysteine endopeptidase (CysEP) with a C-terminal KDEL endoplasmic reticulum retention signal. CysEP is a member of a unique group of papain-type cysteine peptidases found specifically in senescing and ricinosome-containing tissues. During seed development in the castor oil plant (Ricinus communis L.), the cells of the nucellus are killed as the major seed storage organ, the cellular endosperm, expands and begins to accumulate reserves. The destruction of the maternal seed tissues is a developmentally programmed cell death. Terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling revealed that nuclear DNA fragmentation occurs in the nucellar cells adjacent to the expanding endosperm. These cells exhibit ultrastructural features consistent with programmed cell death, including vesiculation of the cytosol, development of irregularly shaped nuclei, vacuolar collapse, and shrinkage of the cytoplasm. Ricinosomes containing the CysEP were identified in the nucellar cells by light and electron microscopy and immunocytochemistry. Both proCysEP and mature CysEP are present in protein extracts of the nucellar tissues during seed development. Upon collapse of the nucellar cells, the content of the ricinosomes is released into the cytoplasm, where the activated CysEP digests the remaining proteinaceous cellular debris. Digestion products of the nucellar cells are presumed taken up by the outermost cells of the endosperm, which have labyrinthine ingrowths of the outer walls typical of transfer cells.

The 2.0 A crystal structure and substrate specificity of the KDEL-tailed cysteine endopeptidase functioning in programmed cell death of Ricinus communis endosperm.

In the senescing endosperm of germinating castor bean (Ricinus communis) a special organelle (the ricinosome) releases a papain-type cysteine endopeptidase (CysEP) during the final stages of cellular disintegration. Protein cleavage sites for the Ricinus CysEP were determined with fluorogenic peptides (Abz-Xaa-Arg-/-Gln-Gln-Tyr(NO2)-Asp). The highest kcat/Km values were obtained with neutral amino acid residues with large aliphatic and non-polar (Leu, Val, Ile, Met) or aromatic (Phe, Tyr, Trp) side-chains. A second series (Abz-Leu-Xaa-/Gln-Pro-Tyr(NO2)-Asp) was evaluated. Based on these results, the covalent binding inhibitor H-D-Val-Leu-Lys-chloromethylketone (CMK) was chosen as substrate analogue for replacement in the catalytic site. Unusually, CysEP cleaved beta-casein N and C-terminal to the amino acid proline. CysEP was crystallized, its structure was solved by molecular replacement at 2.0 A resolution and refined to a R-factor of 18.1% (Rfree=22.6%). The polypeptide chain folds as in papain into two domains divided by the active site cleft, an elongated surface depression harboring the active site. The non-primed specificity subsites of the proteinase are clearly defined by the H-D-Val-Leu-Lys-CMK-inhibitor covalently bound to the active site. The absence of the occluding loop, which blocks the active site of exopeptidases at the C-terminal side of the scissile bond, identifies CysEP as an endopeptidase. The more open pocket of the Ricinus CysEP correlates with the extended variety of substrate amino acid residues accommodated by this enzyme, including even proline at the P1 and P1' positions. This may allow the enzyme to attack a greater variety of proteins during programmed cell death.