229 resultados para Single-molecule Microscopy


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Introduction: The ability to regulate joint stiffness and coordinate movement during landing when impaired by muscle fatigue has important implications for knee function. Unfortunately, the literature examining fatigue effects on landing mechanics suffers from a lack of consensus. Inconsistent results can be attributed to variable fatigue models, as well as grouping variable responses between individuals when statistically detecting differences between conditions. There remains a need to examine fatigue effects on knee function during landing with attention to these methodological limitations. Aim: The purpose of this study therefore, was to examine the effects of isokinetic fatigue on pre-impact muscle activity and post-impact knee mechanics during landing using singlesubject analysis. Methodology: Sixteen male university students (22.6+3.2 yrs; 1.78+0.07 m; 75.7+6.3 kg) performed maximal concentric and eccentric knee extensions in a reciprocal manner on an isokinetic dynamometer and step-landing trials on 2 occasions. On the first occasion each participant performed 20 step-landing trials from a knee-high platform followed by 75 maximal contractions on the isokinetic dynamometer. The isokinetic data was used to calculate the operational definition of fatigue. On the second occasion, with a minimum rest of 14 days, participants performed 2 sets of 20 step landing trials, followed by isokinetic exercise until the operational definition of fatigue was met and a final post-fatigue set of 20 step-landing trials. Results: Single-subject analyses revealed that isokinetic fatigue of the quadriceps induced variable responses in pre impact activation of knee extensors and flexors (frequency, onset timing and amplitude) and post-impact knee mechanics(stiffness and coordination). In general however, isokinetic fatigue induced sig nificant (p<0.05) reductions in quadriceps activation frequency, delayed onset and increased amplitude. In addition, knee stiffness was significantly (p<0.05) increased in some individuals, as well as impaired sagittal coordination. Conclusions: Pre impact activation and post-impact mechanics were adjusted in patterns that were unique to the individual, which could not be identified using traditional group-based statistical analysis. The results suggested that individuals optimised knee function differently to satisfy competing demands, such as minimising energy expenditure, as well as maximising joint stability and sensory information.

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Introduction: Evidence concerning the alteration of knee function during landing suffers from a lack of consensus. This uncertainty can be attributed to methodological flaws, particularly in relation to the statistical analysis of variable human movement data. Aim: The aim of this study was to compare single-subject and group analysis in quantifying alterations in the magnitude and within-participant variability of knee mechanics during a step landing task. Methods: A group of healthy men (N = 12) stepped-down from a knee-high platform for 60 consecutive trials, each trial separated by a 1-minute rest. The magnitude and within-participant variability of sagittal knee stiffness and coordination of the landing leg during the immediate postimpact period were evaluated. Coordination of the knee was quantified in the sagittal plane by calculating the mean absolute relative phase of sagittal shank and thigh motion (MARP1) and between knee rotation and knee flexion (MARP2). Changes across trials were compared between both group and single-subject statistical analyses. Results: The group analysis detected significant reductions in MARP1 magnitude. However, the single-subject analyses detected changes in all dependent variables, which included increases in variability with task repetition. Between-individual variation was also present in the timing, size and direction of alterations to task repetition. Conclusion: The results have important implications for the interpretation of existing information regarding the adaptation of knee mechanics to interventions such as fatigue, footwear or landing height. It is proposed that a familiarisation session be incorporated in future experiments on a single-subject basis prior to an intervention.

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This article offers a critical exploration of the concept of resilience, which is largely conceptualized in the literature as an extraordinary atypical personal ability to revert or ‘bounce back’ to a point of equilibrium despite significant adversity. While resilience has been explored in a range of contexts, there is little recognition of resilience as a social process arising from mundane practices of everyday life and situated in person -environment interactions. Based on an ethnographic study among single refugee women with children in Brisbane, Australia, the women’s stories on navigating everyday tensions and opportunities revealed how resilience was a process operating inter-subjectively in the social spaces connecting them to their environment. Far beyond the simplistic binaries of resilience versus non-resilient, we concern ourselves here with the everyday processual, person environment nature of the concept. We argue that more attention should be paid to day-to-day pathways through which resilience outcomes are achieved, and that this has important implications for refugee mental health practice frameworks.

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Background Bactrocera dorsalis s.s. is a pestiferous tephritid fruit fly distributed from Pakistan to the Pacific, with the Thai/Malay peninsula its southern limit. Sister pest taxa, B. papayae and B. philippinensis, occur in the southeast Asian archipelago and the Philippines, respectively. The relationship among these species is unclear due to their high molecular and morphological similarity. This study analysed population structure of these three species within a southeast Asian biogeographical context to assess potential dispersal patterns and the validity of their current taxonomic status. Results Geometric morphometric results generated from 15 landmarks for wings of 169 flies revealed significant differences in wing shape between almost all sites following canonical variate analysis. For the combined data set there was a greater isolation-by-distance (IBD) effect under a ‘non-Euclidean’ scenario which used geographical distances within a biogeographical ‘Sundaland context’ (r2 = 0.772, P < 0.0001) as compared to a ‘Euclidean’ scenario for which direct geographic distances between sample sites was used (r2 = 0.217, P < 0.01). COI sequence data were obtained for 156 individuals and yielded 83 unique haplotypes with no correlation to current taxonomic designations via a minimum spanning network. BEAST analysis provided a root age and location of 540kya in northern Thailand, with migration of B. dorsalis s.l. into Malaysia 470kya and Sumatra 270kya. Two migration events into the Philippines are inferred. Sequence data revealed a weak but significant IBD effect under the ‘non-Euclidean’ scenario (r2 = 0.110, P < 0.05), with no historical migration evident between Taiwan and the Philippines. Results are consistent with those expected at the intra-specific level. Conclusions Bactrocera dorsalis s.s., B. papayae and B. philippinensis likely represent one species structured around the South China Sea, having migrated from northern Thailand into the southeast Asian archipelago and across into the Philippines. No migration is apparent between the Philippines and Taiwan. This information has implications for quarantine, trade and pest management.

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Molecular dynamics (MD) simulations have been employed to investigate the single-crystal Si properties with different pre-existing cavities under nanoindentation. Cavities with different radii and positions have been considered. It is found that pre-existing cavities in the Si substrate would obviously influence the mechanical properties of Si under nanoindentation. Furthermore, pre-existing cavities would absorb part of the strain energy during loading and then release during unloading. It would decrease plastic deformation to the substrate. Particularly, the larger of the cavity or the nearer of the cavity to the substrate’s top surface, the larger decrease of Young’s modulus and hardness is usually observed. Just as expected, the larger offset of the cavity in the lateral direction, the less influence is usually seen.

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Background: Pre-participation screening is commonly used to measure and assess potential intrinsic injury risk. The single leg squat is one such clinical screening measure used to assess lumbopelvic stability and associated intrinsic injury risk. With the addition of a decline board, the single leg decline squat (SLDS) has been shown to reduce ankle dorsiflexion restrictions and allowed greater sagittal plane movement of the hip and knee. On this basis, the SLDS has been employed in the Cricket Australia physiotherapy screening protocols as a measure of lumbopelvic control in the place of the more traditional single leg flat squat (SLFS). Previous research has failed to demonstrate which squatting technique allows for a more comprehensive assessment of lumbopelvic stability. Tenuous links are drawn between kinematics and hip strength measures within the literature for the SLS. Formal evaluation of subjective screening methods has also been suggested within the literature. Purpose: This study had several focal points namely 1) to compare the kinematic differences between the two single leg squatting conditions, primarily the five key kinematic variables fundamental to subjectively assess lumbopelvic stability; 2) determine the effect of ankle dorsiflexion range of motion has on squat kinematics in the two squat techniques; 3) examine the association between key kinematics and subjective physiotherapists’ assessment; and finally 4) explore the association between key kinematics and hip strength. Methods: Nineteen (n=19) subjects performed five SLDS and five SLFS on each leg while being filmed by an 8 camera motion analysis system. Four hip strength measures (internal/external rotation and abd/adduction) and ankle dorsiflexion range of motion were measured using a hand held dynamometer and a goniometer respectively on 16 of these subjects. The same 16 participants were subjectively assessed by an experienced physiotherapist for lumbopelvic stability. Paired samples t-tests were performed on the five predetermined kinematic variables to assess the differences between squat conditions. A Bonferroni correction for multiple comparisons was used which adjusted the significance value to p = 0.005 for the paired t-tests. Linear regressions were used to assess the relationship between kinematics, ankle range of motion and hip strength measures. Bivariate correlations between hip strength measures and kinematics and pelvic obliquity were employed to investigate any possible relationships. Results: 1) Significant kinematic differences between squats were observed in dominant (D) and non-dominant (ND) end of range hip external rotation (ND p = <0.001; D p = 0.004) and hip adduction kinematics (ND p = <0.001; D p = <0.001). With the mean angle, only the non-dominant leg observed significant differences in hip adduction (p = 0.001) and hip external rotation (p = <0.001); 2) Significant linear relationships were observed between clinical measures of ankle dorsiflexion and sagittal plane kinematic namely SLFS dominant ankle (p = 0.006; R2 = .429), SLFS non-dominant knee (p = 0.015; R2 = .352) and SLFS non-dominant ankle (p = 0.027; R2 = .305) kinematics. Only the dominant ankle (p = 0.020; R2 = .331) was found to have a relationship with the decline squat. 3) Strength measures had tenuous associations with the subjective assessments of lumbopelvic stability with no significant relationships being observed. 4) For the non-dominant leg, external rotation strength and abduction strength were found to be significantly correlated with hip rotation kinematics (Newtons r = 0.458 p = 0.049; Normalised for bodyweight: r = 0.469; p = 0.043) and pelvic obliquity (normalised for bodyweight: r = 0.498 p = 0.030) respectively for the SLFS only. No significant relationships were observed in the dominant leg for either squat condition. Some elements of the hip strength screening protocols had linear relationships with kinematics of the lower limb, particularly the sagittal plane movements of the knee and ankle. Strength measures had tenuous associations with the subjective assessments of lumbopelvic stability with no significant relationships being observed; Discussion: The key finding of this study illustrated that kinematic differences can occur at the hip without significant kinematic differences at the knee as a result of the introduction of a decline board. Further observations reinforce the role of limited ankle dorsiflexion range of motion on sagittal plane movement of the hip and knee and in turn multiplanar kinematics of the lower limb. The kinematic differences between conditions have clinical implications for screening protocols that employ frontal plane movement of the knee as a guide for femoral adduction and rotation. Subjects who returned stronger hip strength measurements also appeared to squat deeper as characterised by differences in sagittal plane kinematics of the knee and ankle. Despite the aforementioned findings, the relationship between hip strength and lower limb kinematics remains largely tenuous in the assessment of the lumbopelvic stability using the SLS. The association between kinematics and the subjective measures of lumbopelvic stability also remain tenuous between and within SLS screening protocols. More functional measures of hip strength are needed to further investigate these relationships. Conclusion: The type of SLS (flat or decline) should be taken into account when screening for lumbopelvic stability. Changes to lower limb kinematics, especially around the hip and pelvis, were observed with the introduction of a decline board despite no difference in frontal plane knee movements. Differences in passive ankle dorsiflexion range of motion yielded variations in knee and ankle kinematics during a self-selected single leg squatting task. Clinical implications of removing posterior ankle restraints and using the knee as a guide to illustrate changes at the hip may result in inaccurate screening of lumbopelvic stability. The relationship between sagittal plane lower limb kinematics and hip strength may illustrate that self-selected squat depth may presumably be a useful predictor of the lumbopelvic stability. Further research in this area is required.

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The most common software analysis tools available for measuring fluorescence images are for two-dimensional (2D) data that rely on manual settings for inclusion and exclusion of data points, and computer-aided pattern recognition to support the interpretation and findings of the analysis. It has become increasingly important to be able to measure fluorescence images constructed from three-dimensional (3D) datasets in order to be able to capture the complexity of cellular dynamics and understand the basis of cellular plasticity within biological systems. Sophisticated microscopy instruments have permitted the visualization of 3D fluorescence images through the acquisition of multispectral fluorescence images and powerful analytical software that reconstructs the images from confocal stacks that then provide a 3D representation of the collected 2D images. Advanced design-based stereology methods have progressed from the approximation and assumptions of the original model-based stereology(1) even in complex tissue sections(2). Despite these scientific advances in microscopy, a need remains for an automated analytic method that fully exploits the intrinsic 3D data to allow for the analysis and quantification of the complex changes in cell morphology, protein localization and receptor trafficking. Current techniques available to quantify fluorescence images include Meta-Morph (Molecular Devices, Sunnyvale, CA) and Image J (NIH) which provide manual analysis. Imaris (Andor Technology, Belfast, Northern Ireland) software provides the feature MeasurementPro, which allows the manual creation of measurement points that can be placed in a volume image or drawn on a series of 2D slices to create a 3D object. This method is useful for single-click point measurements to measure a line distance between two objects or to create a polygon that encloses a region of interest, but it is difficult to apply to complex cellular network structures. Filament Tracer (Andor) allows automatic detection of the 3D neuronal filament-like however, this module has been developed to measure defined structures such as neurons, which are comprised of dendrites, axons and spines (tree-like structure). This module has been ingeniously utilized to make morphological measurements to non-neuronal cells(3), however, the output data provide information of an extended cellular network by using a software that depends on a defined cell shape rather than being an amorphous-shaped cellular model. To overcome the issue of analyzing amorphous-shaped cells and making the software more suitable to a biological application, Imaris developed Imaris Cell. This was a scientific project with the Eidgenössische Technische Hochschule, which has been developed to calculate the relationship between cells and organelles. While the software enables the detection of biological constraints, by forcing one nucleus per cell and using cell membranes to segment cells, it cannot be utilized to analyze fluorescence data that are not continuous because ideally it builds cell surface without void spaces. To our knowledge, at present no user-modifiable automated approach that provides morphometric information from 3D fluorescence images has been developed that achieves cellular spatial information of an undefined shape (Figure 1). We have developed an analytical platform using the Imaris core software module and Imaris XT interfaced to MATLAB (Mat Works, Inc.). These tools allow the 3D measurement of cells without a pre-defined shape and with inconsistent fluorescence network components. Furthermore, this method will allow researchers who have extended expertise in biological systems, but not familiarity to computer applications, to perform quantification of morphological changes in cell dynamics.

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Background:  For those in the field of managing diabetic complications, the accurate diagnosis and monitoring of diabetic peripheral neuropathy (DPN) continues to be a challenge. Assessment of sub-basal corneal nerve morphology has recently shown promise as a novel ophthalmic marker for the detection of DPN. Methods:  Two hundred and thirty-one individuals with diabetes with predominantly mild or no neuropathy and 61 controls underwent evaluation of diabetic neuropathy symptom score, neuropathy disability score, testing with 10 g monofilament, quantitative sensory testing (warm, cold, vibration detection) and nerve conduction studies. Corneal nerve fibre length, branch density and tortuosity were measured using corneal confocal microscopy. Differences in corneal nerve morphology between individuals with and without DPN and controls were investigated using analysis of variance and correlations were determined between corneal morphology and established tests of, and risk factors for, DPN. Results:  Corneal nerve fibre length was significantly reduced in diabetic individuals with mild DPN compared with both controls (p < 0.001) and diabetic individuals without DPN (p = 0.012). Corneal nerve branch density was significantly reduced in individuals with mild DPN compared with controls (p = 0.032). Corneal nerve fibre tortuosity did not show significant differences. Corneal nerve fibre length and corneal nerve branch density showed modest correlations to most measures of neuropathy, with the strongest correlations to nerve conduction study parameters (r = 0.15 to 0.25). Corneal nerve fibre tortuosity showed only a weak correlation to the vibration detection threshold. Corneal nerve fibre length was inversely correlated to glycated haemoglobin (r = -0.24) and duration of diabetes (r = -0.20). Conclusion:  Assessment of corneal nerve morphology is a non-invasive, rapid test capable of showing differences between individuals with and without DPN. Corneal nerve fibre length shows the strongest associations with other diagnostic tests of neuropathy and with established risk factors for neuropathy.

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This paper presents a reactive collision avoidance method for small unmanned rotorcraft using spherical image-based visual servoing. Only a single point feature is used to guide the aircraft in a safe spiral like trajectory around the target, whilst a spherical camera model ensures the target always remains visible. A decision strategy to stop the avoidance control is derived based on the properties of spiral like motion, and the effect of accurate range measurements on the control scheme is discussed. We show that using a poor range estimate does not significantly degrade the collision avoidance performance, thus relaxing the need for accurate range measurements. We present simulated and experimental results using a small quad rotor to validate the approach.

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Recombinant human papillomavirus (HPV) virus-like particles (VLPs) made from the major capsid protein L1 are promising vaccine candidates for use as vaccines against genital and other HPV infections, and particularly against HPV-16. However, HPV-16 genotype variants have different binding affinities for neutralising mouse Mabs raised against HPV-16 L1 VLPs. This paper analyses, using a panel of well-characterised Mabs, the effects on the antigenicity of various C- and N-terminal deletants of HPV-16 L1 made in insect cells via recombinant baculovirus, of an A → T mutation at residue 266 (A266T), and of a C → G mutation at conserved position 428 (C428G). The effects of these changes on assembly of the variant L1s were studied by electron microscopy. Binding of Mab H16:E70 to A266T was reduced by almost half in comparison to wild type L1. Retention of the C-terminal region 428-483 was critical for the binding of conformation-specific Mabs (H16:V5, H16:E70, H16:U4 and H16:9A) whereas deletion of the nuclear localisation signal (NLS) or the C428G mutation or an N-terminal deletion (residues 2-9) did not affect the antigenicity. The N-terminal deletion resulted in a mixed population of 30 and 55 nm VLPs, which differs from the same construct expressed in Escherichia coli, whereas pentamer aggregates resulted from deletion of the 428-465 region or the C428G mutation. The results have implications both for considering use of single-genotype HPV vaccines, and for design of novel second-generation vaccines. © 2006 Elsevier B.V. All rights reserved.

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We used in vivo (biological), in silico (computational structure prediction), and in vitro (model sequence folding) analyses of single-stranded DNA sequences to show that nucleic acid folding conservation is the selective principle behind a high-frequency single-nucleotide reversion observed in a three-nucleotide mutated motif of the Maize streak virus replication associated protein (Rep) gene. In silico and in vitro studies showed that the three-nucleotide mutation adversely affected Rep nucleic acid folding, and that the single-nucleotide reversion [C(601)A] restored wild-type-like folding. In vivo support came from infecting maize with mutant viruses: those with Rep genes containing nucleotide changes predicted to restore a wild-type-like fold [A(601)/G(601)] preferentially accumulated over those predicted to fold differently [C(601)/T(601)], which frequently reverted to A(601) and displaced the original population. We propose that the selection of native nucleic acid folding is an epigenetic effect, which might have broad implications in the evolution of plants and their viruses.