Trunk, Pelvis, Hip, and Knee Kinematics, Hip Strength, and Gluteal Muscle Activation During a Single-Leg Squat in Males and Females With and Without Patellofemoral Pain Syndrome

STUDY DESIGN: Controlled laboratory study using a cross-sectional design. OBJECTIVES: To determine whether there are any differences between the sexes in trunk, pelvis, hip, and knee kinematics, hip strength, and gluteal muscle activation during the performance of a single-leg squat in individuals with patellofemoral pain syndrome (PFPS) and control participants. BACKGROUND: Though there is a greater incidence of PFPS in females, PFPS is also quite common in males. Trunk kinematics may affect hip and knee function; however, there is a lack of studies of the influence of the trunk in individuals with PFPS. METHODS: Eighty subjects were distributed into 4 groups: females with PFPS, female controls, males with PFPS, and male controls. Trunk, pelvis, hip, and knee kinematics and gluteal muscle activation were evaluated during a single-leg squat. Hip abduction and external rotation eccentric strength was measured on an isokinetic dynamometer. Group differences were assessed using a 2-way multivariate analysis of variance (sex by PFPS status). RESULTS: Compared to controls, subjects with PFPS had greater ipsilateral trunk lean (mean +/- SD, 9.3 degrees +/- 5.30 degrees versus 6.7 degrees +/- 3.0 degrees; P = .012), contralateral pelvic drop (10.3 degrees +/- 4.7 degrees versus 7.4 degrees 3.8 degrees; P = .003), hip adduction (14.8 degrees +/- 7.8 degrees versus 10.8 degrees +/- 5.6 degrees; P<.0001), and knee abduction (9.2 degrees +/- 5.0 degrees versus 5.8 degrees +/- 3.4 degrees; P<.0001) when performing a single-leg squat. Subjects with PFPS also had 18% less hip abduction and 17% less hip external rotation strength. Compared to female controls, females with PFPS had more hip internal rotation (P<.05) and less muscle activation of the gluteus medius (P = .017) during the single-leg squat. CONCLUSION: Despite many similarities in findings for males and females with PFPS, there may be specific sex differences that warrant consideration in future studies and when clinically evaluating and treating females with PFPS. J Orthop Sports Phys Ther 2012;42(6):491-501, Epub 8 March 2012. doi:10.2519/jospt.2012.3987

Changes in FDB and soleus muscle activity after a train of stimuli during upright stance

Background: Evidence of self-sustained muscle activation following a brief electrical stimulation has been reported in the literature for certain muscles. Objectives: This report shows that the foot muscle (Flexor Digitorum Brevis - FDB) shows a self-sustained increase in muscle activity during upright stance in some subjects following a train of stimuli to the tibial nerve. Methods: Healthy subjects were requested to stand upright and surface EMG electrodes were placed on the FDB, Soleus and Tibialis Anterior muscles. After background muscle activity (BGA) acquisition, a 50 Hz train of stimuli was applied to the tibial nerve at the popliteal fossa. The root mean square values (RMS) of the BGA and the post-stimulus muscle activation were computed. Results: There was a 13.8% average increase in the FDB muscle EMG amplitude with respect to BGA after the stimulation was turned off. The corresponding post-stimulus Soleus EMG activity decreased by an average of 9.2%. We hypothesize that the sustained contraction observed in the FDB following stimulus may be evidence of persistent inward currents (PIC) generated in FDB spinal motoneurons. The post-stimulus decrease in soleus activity may have occurred due to the action of inhibitory interneurons caused by the PICs, which were triggered by the stimulus train. Conclusions: These sustained post-stimulation changes in postural muscle activity, found in different levels in different subjects, may be part of a set of possible responses that contribute to overall postural control.

A non-cross-bridge, static tension is present in permeabilized skeletal muscle fibers after active force inhibition or actin extraction

Cornachione AS, Rassier DE. A non-cross-bridge, static tension is present in permeabilized skeletal muscle fibers after active force inhibition or actin extraction. Am J Physiol Cell Physiol 302: C566-C574, 2012. First published November 16, 2011; doi: 10.1152/ajpcell.00355.2011.-When activated muscle fibers are stretched, there is a long-lasting increase in the force. This phenomenon, referred to as "residual force enhancement," has characteristics similar to those of the " static tension," a long-lasting increase in force observed when muscles are stretched in the presence of Ca2+ but in the absence of myosin-actin interaction. Independent studies have suggested that these two phenomena have a common mechanism and are caused either by 1) a Ca2+-induced stiffening of titin or by 2) promoting titin binding to actin. In this study, we performed two sets of experiments in which activated fibers (pCa(2+) 4.5) treated with the myosin inhibitor blebbistatin were stretched from 2.7 to 2.8 mu m at a speed of 40 L-o/s, first, after partial extraction of TnC, which inhibits myosin-actin interactions, or, second, after treatment with gelsolin, which leads to the depletion of thin (actin) filaments. We observed that the static tension, directly related with the residual force enhancement, was not changed after treatments that inhibit myosin-actin interactions or that deplete fibers from troponin C and actin filaments. The results suggest that the residual force enhancement is caused by a stiffening of titin upon muscle activation but not with titin binding to actin. This finding indicates the existence of a Ca2+-regulated, titin-based stiffness in skeletal muscles.

Effects of strength and power training on neuromuscular adaptations and jumping movement pattern and performance

Effects of strength and power training on neuromuscular adaptations and jumping movement pattern and performance. J Strength Cond Res 26(12): 3335-3344, 2012-This study aimed at comparing the effects of strength and power training (ST and PT) regimens on neuromuscular adaptations and changes on vertical jump performance, kinetics, and kinematics parameters. Forty physically active men (178.2 +/- 7.0 cm; 75.1 +/- 8.6 kg; 23.6 +/- 3.5 years) with at least 2 years of ST experience were assigned to an ST (n = 14), a PT (n = 14), or a control group (C; n = 12). The training programs were performed during 8 weeks, 3 times per week. Dynamic and isometric maximum strength, cross-sectional area, and muscle activation were assessed before and after the experimental period. Squat jump (SJ) and countermovement jump (CMJ) performance, kinetics, and kinematics parameters were also assessed. Dynamic maximum strength increased similarly (p < 0.05) for the ST (22.8%) and PT (16.6%) groups. The maximum voluntary isometric contraction increased for the ST and PT groups (p < 0.05) in the posttraining assessments. There was a main time effect for muscle fiber cross-sectional area (p < 0.05), but there were no changes in muscle activation. The SJ height increased, after ST and PT, because of a faster concentric phase and a higher rate of force development (p < 0.05). The CMJ height increased only after PT (p < 0.05), but there were no significant changes in its kinetics and kinematics parameters. In conclusion, neuromuscular adaptations were similar between the training groups. The PT seemed more effective than the ST in increasing jumping performance, but neither the ST nor the PT was able to affect the SJ and the CMJ movement pattern (e.g., timing and sequencing of joint extension initiation).

Effect of a rocker non-heeled shoe on EMG and ground reaction forces during gait without previous training

Unstable shoes have been designed to promote "natural instability" and during walking they should simulate barefoot gait, enhancing muscle activity and, thus, attributing an advantage over regular tennis shoes. Recent studies showed that, after special training on the appropriate walking pattern, the use of the Masai Barefoot Technology (MBT) shoe increases muscle activation during walking. Our study presents a comparison of muscle activity as well as horizontal and vertical forces during gait with the MBT, a standard tennis shoe and barefoot walking of healthy individuals without previous training. These variables were compared in 25 female subjects and gait conditions were compared using ANOVA repeated measures (effect size:0.25). Walking with the MBT shoe in this non-instructed condition produced higher vertical forces (first vertical peak and weight acceptance rate) than walking with a standard shoe or walking barefoot, which suggests an increase in the loads received by the musculoskeletal system, especially at heel strike. Walking with the MBT shoe did not increase muscle activity when compared to walking with the standard shoe. The barefoot condition was more effective than the MBT shoe at enhancing muscle activation. Therefore, in healthy individuals, no advantage was found in using the MBT over a standard tennis shoe without a special training period. Further studies using the MBT without any instruction over a longer period are needed to evaluate if the higher loads observed in the present study would return to their baseline values after a period of adaptation, and if the muscle activity would increase over time. (C) 2012 Elsevier B.V. All rights reserved.

Frontal Plane Biomechanics in Males and Females with and without Patellofemoral Pain

NAKAGAWA, T. H., E. T. U. MORIYA, C. D. MACIEL, and F. V. SERRAO. Frontal Plane Biomechanics in Males and Females with and without Patellofemoral Pain. Med. Sci. Sports &ere., Vol. 44, No. 9, pp. 1747-1755, 2012. Purpose: The study's purpose was to compare trunk, pelvis, hip, and knee frontal plane biomechanics in males and females with and without patellofemoral pain syndrome (PFPS) during stepping. Methods: Eighty recreational athletes were equally divided into four groups: female PFPS, female controls, male PFPS, and male controls. Trunk, pelvis, hip, and knee frontal plane kinematics and activation of the gluteus medius were evaluated at 15 degrees, 30 degrees, 45 degrees, and 60 degrees of knee flexion during the downward and upward phases of the stepping task. Isometric hip abductor torque was also evaluated. Results: Females showed increased hip adduction and knee abduction at all knee flexion angles, greater ipsilateral trunk lean and contralateral pelvic drop from 60 degrees of knee flexion till the end of the stepping task (P = 0.027-0.001), diminished hip abductor torque (P < 0.001), and increased gluteus medius activation than males (P = 0.008-0.001). PFPS subjects presented increased knee abduction at all the angles evaluated; greater trunk, pelvis, and hip motion from 45 of knee flexion of the downward phase till the end of the maneuver; and diminished gluteus medius activation at 60 degrees of knee flexion, compared with controls (P = 0.034-0.001). Females with PFPS showed lower hip abductor torque compared with the other groups. Conclusions: Females presented with altered frontal plane biomechanics that may predispose them to knee injury. PFPS subjects showed frontal plane biomechanics that could increase the lateral patellofemoral joint stress at all the angles evaluated and could increase even more from 45 degrees of knee flexion in the downward phase untill the end of the maneuver. Hip abductor strengthening and motor control training should be considered when treating females with PFPS.

Keeping your balance while balancing a cylinder: interaction between postural and voluntary goals

The present study investigated whether postural responses are influenced by the stability constraint of a voluntary, manual task. We also examined how task constraint and first experience (the condition with which the participants started the experiment) influence the kinematic strategies used to simultaneously accomplish a postural response and a voluntary task. Twelve healthy, older adults were perturbed during standing, while holding a tray with a cylinder placed with the flat side down (low constraint, LC) or with the rolling, round side down (high constraint, HC). Central set changed according to the task constraint, as shown by a higher magnitude of both the gastrocnemius and tibialis anterior muscle activation bursts in the HC than in the LC condition. This increase in muscle activation was not reflected, however, in changes in the center of pressure or center of mass displacement. Task constraint influenced the peak shoulder flexion for the voluntary tray task but not the peak hip flexion for the postural task. In contrast, first experience influenced the peak hip flexion but not the peak shoulder flexion. These results suggest an interaction between two separate control mechanisms for automatic postural responses and voluntary stabilization tasks.

EFFECTS OF MUSCULAR STRETCHING AND SEGMENTAL STABILIZATION ON FUNCTIONAL DISABILITY AND PAIN IN PATIENTS WITH CHRONIC LOW BACK PAIN: A RANDOMIZED, CONTROLLED TRIAL

Objective: The purpose of this study was to compare the effects of 2 exercise programs, segmental stabilization exercises (SSEs) and stretching of trunk and hamstrings muscles, on functional disability, pain, and activation of the transversus abdominis muscle (TrA), in individuals with chronic low back pain. Methods: A total of 30 participants were enrolled in this study and randomly assigned to 1 of 2 groups as a function of intervention. In the segmental stabilization group (SS), exercises focused on the TrA and lumbar multifidus muscles, whereas in the stretching group (ST), exercises focused on stretching the erector spinae, hamstrings, and triceps surae. Severity of pain (visual analog scale and McGill pain questionnaire) and functional disability (Oswestry disability questionnaire) and TrA muscle activation capacity (Pressure Biofeedback Unit, or PBU) were compared as a function of intervention. Interventions lasted 6 weeks, and sessions happened twice a week (30 minutes each). Analysis of variance was used for intergroup and intragroup comparisons. Results: As compared with baseline, both treatments were effective in relieving pain and improving disability (P < .001). Those in the SS group had significantly higher gains for all variables. The stretching group did not effectively activate the TrA (P = .94). Conclusion: Both techniques improved pain and reduced disability. In this study, SS was superior to muscular stretching for the measured variables associated with chronic low back pain. (J Manipulative Physiol Ther 2012;35:279-285)

The effect of hip abduction on the EMG activity of vastus medialis obliquus, vastus lateralis longus and vastus lateralis obliquus in healthy subjects

Abstract Study design Controlled laboratory study. Objectives The purposes of this paper were to investigate (d) whether vastus medialis obliquus (VMO), vastus lateralis longus (VLL) and vastus lateralis obliquus (VLO) EMG activity can be influenced by hip abduction performed by healthy subjects. Background Some clinicians contraindicate hip abduction for patellofemoral patients (with) based on the premise that hip abduction could facilitate the VLL muscle activation leading to a VLL and VMO imbalance Methods and measures Twenty-one clinically healthy subjects were involved in the study, 10 women and 11 men (aged X = 23.3 ± 2.9). The EMG signals were collected using a computerized EMG VIKING II, with 8 channels and three pairs of surface electrodes. EMG activity was obtained from MVIC knee extension at 90° of flexion in a seated position and MVIC hip abduction at 0° and 30° with patients in side-lying position with the knee in full extension. The data were normalized in the MVIC knee extension at 50° of flexion in a seated position, and were submitted to ANOVA test with subsequent application of the Bonferroni multiple comparisons analysis test. The level of significance was defined as p ≤ 0.05. Results The VLO muscle demonstrated a similar pattern to the VMO muscle showing higher EMG activity in MVIC knee extension at 90° of flexion compared with MVIC hip abduction at 0° and 30° of abduction for male (p < 0.0007) and MVIC hip abduction at 0° of abduction for female subjects (p < 0.02196). There were no statistically significant differences in the VLL EMG activity among the three sets of exercises tested. Conclusion The results showed that no selective EMG activation was observed when comparison was made between the VMO, VLL and VLO muscles while performing MVIC hip abduction at 0° and 30° of abduction and MVIC knee extension at 90° of flexion in both male and female subjects. Our findings demonstrate that hip abduction do not facilitated VLL and VLO activity in relation to the VMO, however, this study included only healthy subjects performing maximum voluntary isometric contraction contractions, therefore much remains to be discovered by future research

The Effects of Palmitic Acid on Nitric Oxide Production by Rat Skeletal Muscle: Mechanism via Superoxide and iNOS Activation

Background: Increased plasma concentrations of free fatty acids (FFA) can lead to insulin resistance in skeletal muscle, impaired effects on mitochondrial function, including uncoupling of oxidative phosphorylation and decrease of endogenous antioxidant defenses. Nitric oxide (NO) is a highly diffusible gas that presents a half-life of 5-10 seconds and is involved in several physiological and pathological conditions. The effects of palmitic acid on nitric oxide (NO) production by rat skeletal muscle cells and the possible mechanism involved were investigated. Methods: Primary cultured rat skeletal muscle cells were treated with palmitic acid and NO production was assessed by nitrite measurement (Griess method) and 4,5-diaminofluorescein diacetate (DAF-2-DA) assay. Nuclear factor-kappa B (NF-kappa B) activation was evaluated by electrophoretic mobility shift assay and iNOS protein content by western blotting. Results: Palmitic acid treatment increased nitric oxide production. This effect was abolished by treatment with NOS inhibitors, L-nitro-arginine (LNA) and L-nitro-arginine methyl esther (L-NAME). NF-kappa B activation and iNOS content were increased due to palmitic acid treatment. The participation of superoxide on nitric oxide production was investigated by incubating the cells with DAF-2-DA in the presence or absence of palmitic acid, a superoxide generator system (X-XO), a mixture of NOS inhibitors and SOD-PEG (superoxide dismutase linked to polyethylene glycol). Palmitic acid and X-XO system increased NO production and this effect was abolished when cells were treated with NOS inhibitors and also with SOD-PEG. Conclusions: In summary, palmitic acid stimulates NO production in cultured skeletal muscle cells through production of superoxide, nuclear factor-kappa B activation and increase of iNOS protein content. Copyright (C) 2012 S. Karger AG, Basel

Protein Disulfide Isomerase Is Required for Platelet-derived Growth Factor-induced Vascular Smooth Muscle Cell Migration, Nox1 NADPH Oxidase Expression, and RhoGTPase Activation

Vascular Smooth Muscle Cell (VSMC) migration into vessel neointima is a therapeutic target for atherosclerosis and postinjury restenosis. Nox1 NADPH oxidase-derived oxidants synergize with growth factors to support VSMC migration. We previously described the interaction between NADPH oxidases and the endoplasmic reticulum redox chaperone protein disulfide isomerase (PDI) in many cell types. However, physiological implications, as well as mechanisms of such association, are yet unclear. We show here that platelet-derived growth factor (PDGF) promoted subcellular redistribution of PDI concomitant to Nox1-dependent reactive oxygen species production and that siRNA-mediated PDI silencing inhibited such reactive oxygen species production, while nearly totally suppressing the increase in Nox1 expression, with no change in Nox4. Furthermore, PDI silencing inhibited PDGF-induced VSMC migration assessed by distinct methods, whereas PDI overexpression increased spontaneous basal VSMC migration. To address possible mechanisms of PDI effects, we searched for PDI interactome by systems biology analysis of physical protein-protein interaction networks, which indicated convergence with small GTPases and their regulator RhoGDI. PDI silencing decreased PDGF-induced Rac1 and RhoA activities, without changing their expression. PDI co-immunoprecipitated with RhoGDI at base line, whereas such association was decreased after PDGF. Also, PDI co-immunoprecipitated with Rac1 and RhoA in a PDGF-independent way and displayed detectable spots of perinuclear co-localization with Rac1 and RhoGDI. Moreover, PDI silencing promoted strong cytoskeletal changes: disorganization of stress fibers, decreased number of focal adhesions, and reduced number of RhoGDI-containing vesicular recycling adhesion structures. Overall, these data suggest that PDI is required to support Nox1/redox and GTPase-dependent VSMC migration.

Lufaxin, a Novel Factor Xa Inhibitor From the Salivary Gland of the Sand Fly Lutzomyia longipalpis Blocks Protease-Activated Receptor 2 Activation and Inhibits Inflammation and Thrombosis In Vivo

Objective-Blood-sucking arthropods' salivary glands contain a remarkable diversity of antihemostatics. The aim of the present study was to identify the unique salivary anticoagulant of the sand fly Lutzomyia longipalpis, which remained elusive for decades. Methods and Results-Several L. longipalpis salivary proteins were expressed in human embryonic kidney 293 cells and screened for inhibition of blood coagulation. A novel 32.4-kDa molecule, named Lufaxin, was identified as a slow, tight, noncompetitive, and reversible inhibitor of factor Xa (FXa). Notably, Lufaxin's primary sequence does not share similarity to any physiological or salivary inhibitors of coagulation reported to date. Lufaxin is specific for FXa and does not interact with FX, Dansyl-Glu-Gly-Arg-FXa, or 15 other enzymes. In addition, Lufaxin blocks prothrombinase and increases both prothrombin time and activated partial thromboplastin time. Surface plasmon resonance experiments revealed that FXa binds Lufaxin with an equilibrium constant approximate to 3 nM, and isothermal titration calorimetry determined a stoichiometry of 1:1. Lufaxin also prevents protease-activated receptor 2 activation by FXa in the MDA-MB-231 cell line and abrogates edema formation triggered by injection of FXa in the paw of mice. Moreover, Lufaxin prevents FeCl3-induced carotid artery thrombus formation and prolongs activated partial thromboplastin time ex vivo, implying that it works as an anticoagulant in vivo. Finally, salivary gland of sand flies was found to inhibit FXa and to interact with the enzyme. Conclusion-Lufaxin belongs to a novel family of slow-tight FXa inhibitors, which display antithrombotic and anti-inflammatory activities. It is a useful tool to understand FXa structural features and its role in prohemostatic and proinflammatory events. (Arterioscler Thromb Vasc Biol. 2012;32:2185-2196.)

Assessing Pathogenicity for Novel Mutation/Sequence Variants: The Value of Healthy Older Individuals

Improvement in DNA technology is increasingly revealing unexpected/unknown mutations in healthy persons and generating anxiety due to their still unknown health consequences. We report a 44-year-old healthy father of a 10-year-old daughter with bilateral coloboma and hearing loss, but without muscle weakness, in whom a whole-genome CGH revealed a deletion of exons 38-44 in the dystrophin gene. This mutation was inherited from her asymptomatic father, who was further clinically and molecularly evaluated for prognosis and genetic counseling (GC). This deletion was never identified by us in 982 Duchenne/Becker patients. To assess whether the present case represents a rare case of non-penetrance, and aiming to obtain more information for prognosis and GC, we suggested that healthy older relatives submit their DNA for analysis, to which several complied. Mutation analysis revealed that his mother, brother, and 56-year-old maternal uncle also carry the 38-44 deletion, suggesting it an unlikely cause of muscle weakness. Genome sequencing will disclose mutations and variants whose health impact are still unknown, raising important problems in interpreting results, defining prognosis, and discussing GC. We suggest that, in addition to family history, keeping the DNA of older relatives could be very informative, in particular for those interested in having their genome sequenced.

Lack of beta(2)-AR improves exercise capacity and skeletal muscle oxidative phenotype in mice

beta(2)-adrenergic receptor (beta(2)-AR) agonists have been used as ergogenics by athletes involved in training for strength and power in order to increase the muscle mass. Even though anabolic effects of beta(2)-AR activation are highly recognized, less is known about the impact of beta(2)-AR in endurance capacity. We presently used mice lacking beta(2)-AR [beta(2)-knockout (beta(2) KO)] to investigate the role of beta(2)-AR on exercise capacity and skeletal muscle metabolism and phenotype. beta(2) KO mice and their wild-type controls (WT) were studied. Exercise tolerance, skeletal muscle fiber typing, capillary-to-fiber ratio, citrate synthase activity and glycogen content were evaluated. When compared with WT, beta 2KO mice displayed increased exercise capacity (61%) associated with higher percentage of oxidative fibers (21% and 129% of increase in soleus and plantaris muscles, respectively) and capillarity (31% and 20% of increase in soleus and plantaris muscles, respectively). In addition, beta 2KO mice presented increased skeletal muscle citrate synthase activity (10%) and succinate dehydrogenase staining. Likewise, glycogen content (53%) and periodic acid-Schiff staining (glycogen staining) were also increased in beta 2KO skeletal muscle. Altogether, these data provide evidence that disruption of beta(2)AR improves oxidative metabolism in skeletal muscle of beta 2KO mice and this is associated with increased exercise capacity.

Immunological parameters in elderly women: Correlations with aerobic power, muscle strength and mood state

Purpose: Our objective was to relate immunological data for healthy but sedentary elderly women to aerobic power, strength, and mood state. Methods: We measured peak aerobic power and one-repetition maximum strength along with mood (depression and fatigue), quality of life and carbohydrate intake on 42 women aged 60-77 years. Standard immunological techniques determined natural killer cell count and cytotoxic activity (NKCA), proliferative responses to phytohemaglutinin and OKT3, various lymphocyte subpopulations (CD3(+), CD3(-)CD19(+), CD56(+), CD4(+), CD8(+), CD56(dim) and CD56(bright)), and markers of activation, maturation, down-regulation and susceptibility to apoptosis (CD25(+), CD28(+), CD45RA(+), CD45RO(+), CD69(+), CD95(+), HLA-DR+). Results: Correlations of immune parameters with aerobic power and strength were very similar for absolute and relative immunological data. In the group as a whole, the only correlation with aerobic power was -0.35 (relative CD4(+)CD69(+) count), but in subjects with values <22.6 mL kg(-1) min(-1) correlations ranged from -0.57 (relative CD4(+)CD45RO(+)) to 0.92 (absolute CD56(dim)HLA-DR+). In terms of muscle strength, univariate correlation coefficients ranged from -0.34 (relative and absolute CD3(+)CD4(+)CD8(+)) to +0.48 (absolute CD3(+)HLA-DR+.) and +0.50 (absolute CD8(+)CD45RA(+)CD45RO(+)). Neither NKCA nor lymphocyte proliferation were correlated with aerobic power or muscle strength. Although mood state and quality of life can sometimes be influenced by an individual's fitness level, our multivariate analyses suggested that depression, fatigue and quality of life were more important determinants of immune profile than our fitness measures. Conclusions: Psychological changes associated with aging may have a substantial adverse effect upon the immune system, and immunological function may be enhanced more by addressing these issues than by focusing upon aerobic or resistance training. (C) 2012 Elsevier Inc. All rights reserved.