Dynamic stability of individuals with transtibial amputation walking in destabilizing environments

Lower limb amputation substantially disrupts motor and proprioceptive function. People with lower limb amputation experience considerable impairments in walking ability, including increased fall risk. Understanding the biomechanical aspects of the gait of these patients is crucial in improving their gait function and their quality of life. In the present study, 9 persons with unilateral transtibial amputation and 13 able-bodied controls walked on a large treadmill in a Computer Assisted Rehabilitation Environment (CAREN). While walking, subjects were either not perturbed, or were perturbed either by continuous mediolateral platform movements or by continuous mediolateral movements of the visual scene. Means and standard deviations of both step lengths and step widths increased significantly during both perturbation conditions (all p<0.001) for both groups. Measures of variability, local and orbital dynamic stability of trunk movements likewise exhibited large and highly significant increases during both perturbation conditions (all p<0.001) for both groups. Patients with amputation exhibited greater step width variability (p=0.01) and greater trunk movement variability (p=0.04) during platform perturbations, but did not exhibit greater local or orbital instability than healthy controls for either perturbation conditions. Our findings suggest that, in the absence of other co-morbidities, patients with unilateral transtibial amputation appear to retain sufficient sensory and motor function to maintain overall upper body stability during walking, even when substantially challenged. Additionally, these patients did not appear to rely more heavily on visual feedback to maintain trunk stability during these walking tasks.     

Margins of stability in young adults with traumatic transtibial amputation walking in destabilizing environments

Understanding how lower-limb amputation affects walking stability, specifically in destabilizing environments, is essential for developing effective interventions to prevent falls. This study quantified mediolateral margins of stability (MOS) and MOS sub-components in young individuals with traumatic unilateral transtibial amputation (TTA) and young able-bodied individuals (AB). Thirteen AB and nine TTA completed five 3-min walking trials in a Computer Assisted Rehabilitation ENvironment (CAREN) system under each of three test conditions: no perturbations, pseudo-random mediolateral translations of the platform, and pseudo-random mediolateral translations of the visual field. Compared to the unperturbed trials, TTA exhibited increased mean MOS and MOS variability during platform and visual field perturbations (p<0.010). AB exhibited increased mean MOS during visual field perturbations and increased MOS variability during both platform and visual field perturbations (p<0.050). During platform perturbations, TTA exhibited significantly greater values than AB for mean MOS (p<0.050) and MOS variability (p<0.050); variability of the lateral distance between the center of mass (COM) and base of support at initial contact (p<0.005); mean and variability of the range of COM motion (p<0.010); and variability of COM peak velocity (p<0.050). As determined by mean MOS and MOS variability, young and otherwise healthy individuals with transtibial amputation achieved lateral stability similar to that of their able-bodied counterparts during unperturbed and visually-perturbed walking. However, based on mean and variability of MOS, unilateral transtibial amputation was shown to have affected lateral walking stability during platform perturbations.     

Dynamic margins of stability during human walking in destabilizing environments

Understanding how humans maintain stability when walking, particularly when exposed to perturbations, is key to preventing falls. Here, we quantified how imposing continuous, pseudorandom anterior-posterior (AP) and mediolateral (ML) oscillations affected the control of dynamic walking stability. Twelve subjects completed five 3-minute walking trials in the Computer Assisted Rehabilitation ENvironment (CAREN) system under each of 5 conditions: no perturbation (NOP), AP platform (APP) or visual (APV) or ML platform (MLP) or visual (MLV) oscillations. We computed AP and ML margins of stability (MOS) for each trial. Mean MOS(ml) were consistently slightly larger during all perturbation conditions than during NOP (p≤0.038). Mean MOS(ap) for the APP, MLP and MLV oscillations were significantly smaller than during NOP (p<0.0005). Variability of both MOS(ap) and MOS(ml) was significantly greater during the MLP and MLV oscillations than during NOP (p<0.0005). We also directly quantified how the MOS on any given step affected the MOS on the following step using first-return plots. There were significant changes in step-to-step MOS(ml) dynamics between experimental conditions (p<0.0005). These changes suggested that subjects may have been trying to control foot placement, and consequently stability, during the perturbation conditions. Quantifying step-to-step changes in margins of dynamic stability may be more useful than mean MOS in assessing how individuals control walking stability.     

Dynamic stability of human walking in visually and mechanically destabilizing environments

Understanding how humans remain stable during challenging locomotor activities is critical to developing effective tests to diagnose patients with increased fall risk. This study determined if different continuous low-amplitude perturbations would induce specific measureable changes in measures of dynamic stability during walking. We applied continuous pseudo-random oscillations of either the visual scene or support surface in either the anterior-posterior or mediolateral directions to subjects walking in a virtual environment with speed-matched optic flow. Floquet multipliers and short-term local divergence exponents both increased (indicating greater instability) during perturbed walking. These responses were generally much stronger for body movements occurring in the same directions as the applied perturbations. Likewise, subjects were more sensitive to both visual and mechanical perturbations applied in the mediolateral direction than to those applied in the anterior-posterior direction, consistent with previous experiments and theoretical predictions. These responses were likewise consistent with subjects' anecdotal perceptions of which perturbation conditions were most challenging. Contrary to the Floquet multipliers and short-term local divergence exponents, which both increased, long-term local divergence exponents decreased during perturbed walking. However, this was consistent with specific changes in the mean log divergence curves, which indicated that subjects' movements reached their maximum local divergence limits more quickly during perturbed walking. Overall, the Floquet multipliers were less sensitive, but reflected greater specificity in their responses to the different perturbation conditions. Conversely, the short-term local divergence exponents exhibited less specificity in their responses, but were more sensitive measures of instability in general.     

A general model for estimating lower extremity inertial properties of individuals with transtibial amputation

Lower extremity joint moment magnitudes during swing are dependent on the inertial properties of the prosthesis and residual limb of individuals with transtibial amputation (TTA). Often, intact limb inertial properties (INTACT) are used for prosthetic limb values in an inverse dynamics model even though these values overestimate the amputated limb’s inertial properties. The purpose of this study was to use subject-specific (SPECIFIC) measures of prosthesis inertial properties to generate a general model (GENERAL) for estimating TTA prosthesis inertial properties. Subject-specific mass, center of mass, and moment of inertia were determined for the shank and foot segments of the prosthesis (n = 11) using an oscillation technique and reaction board. The GENERAL model was derived from the means of the SPECIFIC model. Mass and segment lengths are required GENERAL model inputs. Comparisons of segment inertial properties and joint moments during walking were made using three inertial models (unique sample; n = 9): (1) SPECIFIC, (2) GENERAL, and (3) INTACT. Prosthetic shank inertial properties were significantly smaller with the SPECIFIC and GENERAL model than the INTACT model, but the SPECIFIC and GENERAL model did not statistically differ. Peak knee and hip joint moments during swing were significantly smaller for the SPECIFIC and GENERAL model compared with the INTACT model and were not significantly different between SPECIFIC and GENERAL models. When subject-specific measures are unavailable, using the GENERAL model produces a better estimate of prosthetic side inertial properties resulting in more accurate joint moment measurements for individuals with TTA than the INTACT model.     

Effect of prosthetic alignment changes on socket reaction moment impulse during walking in transtibial amputees

The alignment of a lower limb prosthesis affects the way load is transferred to the residual limb through the socket, and this load is critically important for the comfort and function of the prosthesis. Both magnitude and duration of the moment are important factors that may affect the residual limb health. Moment impulse is a well-accepted measurement that incorporates both factors via moment–time integrals. The aim of this study was to investigate the effect of alignment changes on the socket reaction moment impulse in transtibial prostheses. Ten amputees with transtibial prostheses participated in this study. The socket reaction moment impulse was measured at a self-selected walking speed using a Smart Pyramid™ in 25 alignment conditions, including a nominal alignment (clinically aligned by a prosthetist), as well as angle malalignments of 2°, 4° and 6° (abduction, adduction, extension and flexion) and translation malalignments of 5 mm, 10 mm and 15 mm (lateral, medial, anterior and posterior). The socket reaction moment impulse of the nominal alignment was compared for each condition. The relationship between the alignment and the socket reaction moment impulse was clearly observed in the coronal angle, coronal translation and sagittal translation alignment changes. However, this relationship was not evident in the sagittal angle alignment changes. The results of this study suggested that the socket reaction moment impulse could potentially serve as a valuable parameter to assist the alignment tuning process for transtibial prostheses. Further study is needed to investigate the influence of the socket reaction moment impulse on the residual limb health.     

Two biomechanical strategies for locomotor adaptation to split-belt treadmill walking in subjects with and without transtibial amputation

Locomotor adaptation is commonly studied using split-belt treadmill walking, in which each foot is placed on a belt moving at a different speed. As subjects adapt to split-belt walking, they reduce metabolic power, but the biomechanical mechanism behind this improved efficiency is unknown. Analyzing mechanical work performed by the legs and joints during split-belt adaptation could reveal this mechanism. Because ankle work in the step-to-step transition is more efficient than hip work, we hypothesized that control subjects would reduce hip work on the fast belt and increase ankle work during the step-to-step transition as they adapted. We further hypothesized that subjects with unilateral, trans-tibial amputation would instead increase propulsive work from their intact leg on the slow belt. Control subjects reduced hip work and shifted more ankle work to the step-to-step transition, supporting our hypothesis. Contrary to our second hypothesis, intact leg work, ankle work and hip work in amputees were unchanged during adaptation. Furthermore, all subjects increased collisional energy loss on the fast belt, but did not increase propulsive work. This was possible because subjects moved further backward during fast leg single support in late adaptation than in early adaptation, compensating by reducing backward movement in slow leg single support. In summary, subjects used two strategies to improve mechanical efficiency in split-belt walking adaptation: a CoM displacement strategy that allows for less forward propulsion on the fast belt; and, an ankle timing strategy that allows efficient ankle work in the step-to-step transition to increase while reducing inefficient hip work.     

The effect of walking speed on local dynamic stability is sensitive to calculation methods

Local dynamic stability has been assessed by the short-term local divergence exponent (λ_S), which quantifies the average rate of logarithmic divergence of infinitesimally close trajectories in state space. Both increased and decreased local dynamic stability at faster walking speeds have been reported. This might pertain to methodological differences in calculating λ_S. Therefore, the aim was to test if different calculation methods would induce different effects of walking speed on local dynamic stability. Ten young healthy participants walked on a treadmill at five speeds (60%, 80%, 100%, 120% and 140% of preferred walking speed) for 3 min each, while upper body accelerations in three directions were sampled. From these time-series, λ_S was calculated by three different methods using: (a) a fixed time interval and expressed as logarithmic divergence per stride-time (λ_S−a), (b) a fixed number of strides and expressed as logarithmic divergence per time (λ_S−b) and (c) a fixed number of strides and expressed as logarithmic divergence per stride-time (λ_S−c). Mean preferred walking speed was 1.16±0.09 m/s. There was only a minor effect of walking speed on λ_S−a. λ_S−b increased with increasing walking speed indicating decreased local dynamic stability at faster walking speeds, whereas λ_S−c decreased with increasing walking speed indicating increased local dynamic stability at faster walking speeds. Thus, the effect of walking speed on calculated local dynamic stability was significantly different between methods used to calculate local dynamic stability. Therefore, inferences and comparisons of studies employing λ_S should be made with careful consideration of the calculation method.     

Effect of stable and unstable load carriage on walking gait variability,dynamic stability and muscle activity of older adults

Load carriage perturbs the neuromuscular system, which can be impaired due to ageing. The ability to counteract perturbations is an indicator of neuromuscular function but if the response is insufficient the risk of falls will increase. However, it is unknown how load carriage affects older adults. Fourteen older adults (65 ± 6 years) attended a single visit during which they performed 4 min of walking in 3 conditions, unloaded, stable backpack load and unstable backpack load. During each walking trial, 3-dimensional kinematics of the lower limb and trunk movements and electromyographic activity of 6 lower limb muscles were recorded. The local dynamic stability (local divergence exponents), joint angle variability and spatio-temporal variability were determined along with muscle activation magnitudes. Medio-lateral dynamic stability was lower (p = 0.018) and step width (p = 0.019) and step width variability (p = 0.015) were greater in unstable load walking and step width variability was greater in stable load walking (p = 0.009) compared to unloaded walking. However, there was no effect on joint angle variability. Unstable load carriage increased activity of the Rectus Femoris (p = 0.001) and Soleus (p = 0.043) and stable load carriage increased Rectus Femoris activity (p = 0.006). These results suggest that loaded walking alters the gait of older adults and that unstable load carriage reduces dynamic stability compared to unloaded walking. This can potentially increase the risk of falls, but also offers the potential to use unstable loads as part of fall prevention programmes.     

Control of lateral weight transfer is associated with walking speed in individuals post-stroke

Restoring functional gait speed is an important goal for rehabilitation post-stroke. During walking, transferring of one’s body weight between the limbs and maintaining balance stability are necessary for independent functional gait. Although it is documented that individuals post-stroke commonly have difficulties with performing weight transfer onto their paretic limbs, it remains to be determined if these deficits contributed to slower walking speeds. The primary purpose of this study was to compare the weight transfer characteristics between slow and fast post-stroke ambulators. Participants (N = 36) with chronic post-stroke hemiparesis walked at their comfortable and maximal walking speeds on a treadmill. Participants were stratified into 2 groups based on their comfortable walking speeds (≥0.8 m/s or <0.8 m/s). Minimum body center of mass (COM) to center of pressure (COP) distance, weight transfer timing, step width, lateral foot placement relative to the COM, hip moment, peak vertical and anterior ground reaction forces, and changes in walking speed were analyzed. Results showed that slow walkers walked with a delayed and deficient weight transfer to the paretic limb, lower hip abductor moment, and more lateral paretic limb foot placement relative to the COM compared to fast walkers. In addition, propulsive force and walking speed capacity was related to lateral weight transfer ability. These findings demonstrated that deficits in lateral weight transfer and stability could potentially be one of the limiting factors underlying comfortable walking speeds and a determinant of chronic stroke survivors’ ability to increase walking speed.     

Dynamic gait stability of treadmill versus overground walking in young adults

Treadmill has been broadly used in laboratory and rehabilitation settings for the purpose of facilitating human locomotion analysis and gait training. The objective of this study was to determine whether dynamic gait stability differs or resembles between the two walking conditions (overground vs. treadmill) among young adults. Fifty-four healthy young adults (age: 23.9 ± 4.7 years) participated in this study. Each participant completed five trials of overground walking followed by five trials of treadmill walking at a self-selected speed while their full body kinematics were gathered by a motion capture system. The spatiotemporal gait parameters and dynamic gait stability were compared between the two walking conditions. The results revealed that participants adopted a “cautious gait” on the treadmill compared with over ground in response to the possible inherent challenges to balance imposed by treadmill walking. The cautious gait, which was achieved by walking slower with a shorter step length, less backward leaning trunk, shortened single stance phase, prolonged double stance phase, and more flatfoot landing, ensures the comparable dynamic stability between the two walking conditions. This study could provide insightful information about dynamic gait stability control during treadmill ambulation in young adults.     

Dynamic balance in persons with multiple sclerosis who have a falls history is altered compared to non-fallers and to healthy controls

Around 60% of persons with multiple sclerosis (MS) experience falls, however the dynamic balance differences between those who fall and those who don’t are not well understood. The purpose of this study is to identify distinct biomechanical features of dynamic balance during gait that are different between fallers with MS, non-fallers with MS, and healthy controls. 27 recurrent fallers with MS, 28 persons with MS with no falls history, and 27 healthy controls walked on a treadmill at their preferred speed for 3 min. The variability of trunk accelerations and the average and variability of minimum toe clearance, spatiotemporal parameters, and margin of stability were compared between groups. Fallers with MS exhibited a slower cautious gait compared to non-fallers and healthy controls, but had decreased anterior-posterior margin of stability and minimum toe clearance. Fallers walked with less locally stable and predictable trunk accelerations, and increased variability of step length, stride time, and both anterior-posterior and mediolateral margin of stability compared to non-fallers and healthy controls. The present work provides evidence that within a group of persons with MS, there are gait differences that are influenced by falls history. These differences indicate that in persons with MS who fall, the center of mass is poorly controlled through base of support placement and the foot is closer to the ground during swing phase relative to the non-fallers. These identified biomechanical differences could be used to evaluate dynamic balance in persons with MS and to help improve fall prevention strategies.     

Retention of improvement in gait stability over 14 weeks due to trip-perturbation training is dependent on perturbation dose

Perturbation training is an emerging approach to reduce fall risk in the elderly. This study examined potential differences in retention of improvements in reactive gait stability over 14 weeks resulting from unexpected trip-like gait perturbations. Twenty-four healthy middle-aged adults (41–62 years) were assigned randomly to either a single perturbation group (SINGLE, n = 9) or a group subjected to eight trip-like gait perturbations (MULTIPLE, n = 15). While participants walked on a treadmill a custom-built brake-and-release system was used to unexpectedly apply resistance during swing phase to the lower right limb via an ankle strap. The anteroposterior margin of stability (MoS) was calculated as the difference between the anterior boundary of the base of support and the extrapolated centre of mass at foot touchdown for the perturbed step and the first recovery step during the first and second (MULTIPLE group only) perturbation trials for the initial walking session and retention-test walking 14 weeks later. Group MULTIPLE retained the improvements in reactive gait stability to the perturbations (increased MoS at touchdown for perturbed and first recovery steps; p < 0.01). However, in group SINGLE no differences in MoS were detected after 14 weeks compared to the initial walking session. These findings provide evidence for the requirement of a threshold trip-perturbation dose if adaptive changes in the human neuromotor system over several months, aimed at the improvement in fall-resisting skills, are to occur.     

Ratings of perceived exertion in individuals with varying fitness levels during walking and running

It was the purpose of this investigation to: 1) compare the ratings of perceived exertion (RPEs) in high and low fit individuals when walking and running at comparable exercise intensities and 2) to determine if ventilation (VE) provides a central signal for RPEs. Nine high fit and nine low fit male subjects completed two exercise bouts on a treadmill, one uphill walking and the other level running. Workloads for each bout were set at 90% of each subject's ventilatory threshold (VT) as determined from a graded exercise test. Oxygen consumption (Vo2), heart rate (HR), and VE were all similar between the walk and run trials for the low fit subjects (P greater than 0.05). HR were found to be significantly greater during the walk trial vs. the run trial (P less than 0.05) for the high fit subjects, whereas, VE was significantly greater during the run trial. Oxygen consumption was similar for the high fit subjects during both trials (P greater than 0.05). During the walk and run trials, central (12.1 +/- 1.6 vs. 11.4 +/- 1.5), local (14.0 +/- 1.3 vs. 13.9 +/- 1.1) and overall (12.8 +/- 1.2 vs. 12.4 +/- 1.4) RPEs were not found to be significantly different for the low fit group (P greater than 0.05). In contrast, during the walk vs. the run trial there was a significant increase in central (10.7 +/- 2.0 vs. 9.2 +/- 1.9), local (11.5 +/- 2.0 vs. 9.8 +/- 1.8) and overall (11.2 +/- 2.4 vs. 9.6 +/- 2.3) RPEs for the high fit group (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Magnitude of forward trunk flexion influences upper limb muscular efforts and dynamic postural stability requirements during sitting pivot transfers in individuals with spinal cord injury

The purpose of this study was to investigate the effects of imposing different degrees of forward trunk flexion during sitting pivot transfers on electromyographic activity at the leading and trailing upper limb muscles and on dynamic stability requirements. Thirty-two individuals with a spinal cord injury performed three types of sitting pivot transfers: natural technique, exaggerated forward trunk flexion and upright trunk position. Ground reaction forces, trunk kinematics, and bilateral electromyographic activity of eight upper limb muscles were recorded. Electromyographic data were analyzed using the area under the curve of the muscular utilization ratio. Dynamic stability requirements of sitting pivot transfers were assess using a dynamic equilibrium model. Compared to the natural strategy, significantly greater muscle activities were found for the forward trunk flexion condition at the anterior deltoid and both heads of the pectorialis major, whereas the upright trunk strategy yielded greater muscle activity at the latissimus dorsii and the triceps. The forward flexed condition was found to be more dynamically stable, with a lower stabilizing force, increased area of base of support and greater distance traveled. Thus, transferring with a more forward trunk inclination, even though it increases work of few muscles, may be a beneficial trade-off because increased dynamic stability of this technique and versatility in terms of potential distance of the transfer.     

The concept of margins of stability can be used to better understand a change in obstacle crossing strategy with an increase in age

The purpose of the current study was to use the margins of stability (MoS) to investigate how older adults choose between minimizing the risk of a forward fall when crossing an obstacle and the ease of maintaining forward progression during the steps taken behind the obstacle. In the current study 143 community-dwelling older adults aged between 55 and 83 years old, were divided into three age groups based on tertials of age. All participants were asked to complete five trials of obstacle walking and five trials of normal walking. For the trials of normal walking, the main difference between groups was that MoS at initial contact was lower in the older age groups. For the trials of obstacle crossing the MoS at the instants of obstacle crossing with both the leading and trailing limb became smaller with an increase in age. This result might imply that older people choose to use a strategy during obstacle crossing that results in smaller chance of falling forward if an obstacle was struck. A negative consequence of this more conservative strategy was a smaller MoS at the instants of initial contact after crossing the obstacle, thus a larger chance of a backward fall. These findings provide more insight into the regulation of stability during obstacle crossing and specifically in the differences in strategy between younger and older people, and therefore these results might be used for further research to investigate whether obstacle crossing strategies are trainable in older adults, which could be used as advisory programs aimed at fall prevention and/or engagement in an active lifestyle.     

Dental fluctuating asymmetry in the Gullah: tests of hypotheses regarding developmental stability in deciduous vs. permanent and male vs. female teeth

In this investigation, deciduous teeth (canines, c; first molars, m1; second molars, m2) and their permanent successors (canines, C; first premolars, P1; second premolars, P2) were used to test two related hypotheses about fluctuating asymmetry (FA). First, based on the biology of the developing dentition, it was predicted that deciduous teeth would be more developmentally stable and thus exhibit less dimensional FA than their permanent successors. Second, based on sex differences in tooth development, it was predicted that female canines would have greater developmental stability (less FA) than male canines. Bucco-lingual measurements were made on dental casts from a single Gullah population. Using a repeated-measures study design (n = 3 repeated measures), we tested these hypotheses on sample sizes ranging from 63-82 antimeric pairs. Neither hypothesis was supported by our data. In most cases, Gullah deciduous teeth did not exhibit statistically significantly less FA than their permanent successors; indeed, statistically significant differences were found for only 3 of 12 deciduous vs. permanent contrasts, and in two cases, the deciduous tooth had greater FA. Female mandibular canines exhibited statistically significantly greater FA than those of males, while there was no statistically significant sex difference in FA for the maxillary canine. FA in these Gullah samples is high when compared to Archaic and late prehistoric Ohio Valley Native Americans, consistent with historical and archaeological evidence that environmental stress was relatively higher in the Gullah population. We suggest that when environmental stress in a population is high, the impact of differences in tooth formation time spans and developmental buffering upon FA may be minor relative to the effect of developmental noise.     

基因与环境互作对云南保山烤烟主要潜香型物质的影响

为研究云南保山烤烟不同品种潜香型物质在不同环境条件下的稳定性问题,将云南保山3个烤烟常栽品种(K326、Y87、Y99)种植在3个海拔的2种土壤上,应用AMMI模型对烤烟主要潜香型物质进行基因型(G)、环境(E)和基因型与环境(G×E)互作分析.结果表明： 基因型、环境及其互作对烤烟叶黄素含量、β-胡萝卜素含量、绿原酸含量的影响均达到显著水平;芸香苷含量主要受基因型影响.烤烟主要潜香型物质含量及其稳定性受G×E互作影响显著,叶黄素含量、β-胡萝卜素含量、绿原酸含量随海拔的增加而显著增加,其稳定性也不同程度得到加强;K326适种在中低海拔区,Y87、Y99适种在中高海拔区.
     

Association of the catalytic subunit of aspartate transcarbamoylase with a zinc-containing polypeptide fragment of the regulatory chain leads to increases in thermal stability.

The regulatory enzyme aspartate transcarbamoylase (ATCase), comprising 2 catalytic (C) trimers and 3 regulatory (R) dimers, owes its stability to the manifold interchain interactions among the 12 polypeptide chains. With the availability of a recombinant 70-amino acid zinc-containing polypeptide fragment of the regulatory chain of ATCase, it has become possible to analyze directly the interaction between catalytic and regulatory chains in a complex of simpler structure independent of other interactions such as those between the 2 C trimers, which also contribute to the stability of the holoenzyme. Also, the effect of the interaction between the polypeptide, termed the zinc domain, and the C trimer on the thermal stability and other properties can be measured directly. Differential scanning microcalorimetry experiments demonstrated that the binding of the zinc domain to the C trimer leads to a complex of markedly increased thermal stability. This was shown with a series of mutant forms of the C trimer, which themselves varied greatly in their temperature of denaturation due to single amino acid replacements. With some C trimers, for which tm varied over a range of 30 degrees C due to diverse amino acid substitutions, the elevation of tm resulting from the interaction with the zinc domain was as large as 18 degrees C. The values of tm for a variety of complexes of mutant C trimers and the wild-type zinc domain were similar to those observed when the holoenzymes containing the mutant C trimers were subjected to heat denaturation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Mass spectrometric and chemical stability of the Asp-Pro bond in herpes simplex virus epitope peptides compared with X-Pro bonds of related sequences.

The mass spectrometric analysis of the immunodominant epitope region (273-284) of herpes simplex virus type 1 (HSV-1) glycoprotein D (gD) showed a favoured fission at the Asp-Pro peptide bond. The fast atom bombardment collision induced dissociation (FAB-CID) study of closely related X-Pro peptides documented that neither the length nor the amino acid composition of the peptide has a significant influence on this preferential cleavage. At the same time the DP bond proved to be sensitive to acidic conditions in the course of peptide synthesis. These observations prompted us to compare the chemical and mass spectrometric stability of a new set of nonapeptides related to the 273-284 epitope region of gD, i.e. SALLEDPVG and SALLEXPVG peptides, where X = A, K, I, S, F, E or D, respectively. The chemical stability of these peptides during acidic hydrolysis was investigated by electrospray ionization mass spectrometry (ESI-MS) and the products were identified by ESI-MS and on-line high performance liquid chromatography-mass spectrometry (HPLC-MS). The mass spectrometric fragmentation and bond stability of the untreated peptide samples were also studied using ESI-MS and liquid secondary ion mass spectrometry (LSIMS). Both the chemical hydrolysis and the mass spectrometric fragmentation showed that the Asp-Pro bond could easily be cleaved, while the KP bond proved to be stable under both circumstances. On the other hand, the XP bond (X = A, I, S, F or E) fragmented easily under the mass spectrometric conditions, but was not sensitive to the acidolysis.