The ray projection method: a numerical approach for determining ideal camera placement

Data piloting is important to ensure accurate marker coordinate data and to minimize camera dropout. Camera dropout results when a camera fails to image a marker, which often occurs when markers merge or become occluded. In this article, we present the conceptual framework for a numerical method of determining where video cameras, if placed, would have an occluded or a merged view of the tracking markers. Experimental data are presented to demonstrate the efficacy of the method as a tool to complement existing data piloting procedures.     

Simultaneous measurement of three-dimensional joint kinematics and ligament strains with optical methods

The objective of this study was to assess the precision and accuracy of a nonproprietary, optical three-dimensional (3D) motion analysis system for the simultaneous measurement of soft tissue strains and joint kinematics. The system consisted of two high-resolution digital cameras and software for calculating the 3D coordinates of contrast markers. System precision was assessed by examining the variation in the coordinates of static markers over time. Three-dimensional strain measurement accuracy was assessed by moving contrast markers fixed distances in the field of view and calculating the error in predicted strain. Three-dimensional accuracy for kinematic measurements was assessed by simulating the measurements that are required for recording knee kinematics. The field of view (190 mm) was chosen to allow simultaneous recording of markers for soft tissue strain measurement and knee joint kinematics. Average system precision was between +/-0.004 mm and +/-0.035 mm, depending on marker size and camera angle. Absolute error in strain measurement varied from a minimum of +/-0.025% to a maximum of +/-0.142%, depending on the angle between cameras and the direction of strain with respect to the camera axes. Kinematic accuracy for translations was between +/-0.008 mm and +/-0.034 mm, while rotational accuracy was +/-0.082 deg to +/-0.160 deg. These results demonstrate that simultaneous optical measurement of 3D soft tissue strain and 3D joint kinematics can be performed while achieving excellent accuracy for both sets of measurements.     

Characterisation of a high resolution small field of view portable gamma camera

A handheld, high-resolution small field of view (SFOV) pinhole gamma camera has been characterised using a new set of protocols adapted from standards previously developed for large field of view (LFOV) systems. Parameters investigated include intrinsic and extrinsic spatial resolution, spatial linearity, uniformity, sensitivity, count rate capability and energy resolution. Camera characteristics are compared to some clinical LFOV gamma cameras and also to other SFOV cameras in development.     

An Assessment of the Effectiveness of High Definition Cameras as Remote Monitoring Tools for Dolphin Ecology Studies

Research involving marine mammals often requires costly field programs. This paper assessed whether the benefits of using cameras outweighs the implications of having personnel performing marine mammal detection in the field. The efficacy of video and still cameras to detect Indo-Pacific bottlenose dolphins (Tursiops aduncus) in the Fremantle Harbour (Western Australia) was evaluated, with consideration on how environmental conditions affect detectability. The cameras were set on a tower in the Fremantle Port channel and videos were perused at 1.75 times the normal speed. Images from the cameras were used to estimate position of dolphins at the water’s surface. Dolphin detections ranged from 5.6 m to 463.3 m for the video camera, and from 10.8 m to 347.8 m for the still camera. Detection range showed to be satisfactory when compared to distances at which dolphins would be detected by field observers. The relative effect of environmental conditions on detectability was considered by fitting a Generalised Estimation Equations (GEEs) model with Beaufort, level of glare and their interactions as predictors and a temporal auto-correlation structure. The best fit model indicated level of glare had an effect, with more intense periods of glare corresponding to lower occurrences of observed dolphins. However this effect was not large (-0.264) and the parameter estimate was associated with a large standard error (0.113). The limited field of view was the main restraint in that cameras can be only applied to detections of animals observed rather than counts of individuals. However, the use of cameras was effective for long term monitoring of occurrence of dolphins, outweighing the costs and reducing the health and safety risks to field personal. This study showed that cameras could be effectively implemented onshore for research such as studying changes in habitat use in response to development and construction activities.     

A protocol for monitoring soft tissue motion under compression garments during drop landings

This study used a single-subject design to establish a valid and reliable protocol for monitoring soft tissue motion under compression garments during drop landings. One male participant performed six 40 cm drop landings onto a force platform, in three compression conditions (none, medium high). Five reflective markers placed on the thigh under the compression garment and five over the garment were filmed using two cameras (1000 Hz). Following manual digitisation, marker coordinates were reconstructed and their resultant displacements and maximum change in separation distance between skin and garment markers were calculated. To determine reliability of marker application, 35 markers were attached to the thigh over the high compression garment and filmed. Markers were then removed and re-applied on three occasions; marker separation and distance to thigh centre of gravity were calculated. Results showed similar ground reaction forces during landing trials. Significant reductions in the maximum change in separation distance between markers from no compression to high compression landings were reported. Typical errors in marker movement under and over the garment were 0.1mm in medium and high compression landings. Re-application of markers showed mean typical errors of 1mm in marker separation and <3mm relative to thigh centre of gravity. This paper presents a novel protocol that demonstrates sufficient sensitivity to detect reductions in soft tissue motion during landings in high compression garments compared to no compression. Additionally, markers placed under or over the garment demonstrate low variance in movement, and the protocol reports good reliability in marker re-application.     

Biosafety and risk assessment framework for selectable marker genes in transgenic crop plants: a case of the science not supporting the politics

Selectable marker gene systems are vital for the development of transgenic crops. Since the creation of the first transgenic plants in the early 1980s and their subsequent commercialization worldwide over almost an entire decade, antibiotic and herbicide resistance selectable marker gene systems have been an integral feature of plant genetic modification. Without them, creating transgenic crops is not feasible on purely economic and practical terms. These systems allow the relatively straightforward identification and selection of plants that have stably incorporated not only the marker genes but also genes of interest, for example herbicide tolerance and pest resistance. Bacterial antibiotic resistance genes are also crucial in molecular biology manipulations in the laboratory. An unprecedented debate has accompanied the development and commercialization of transgenic crops. Divergent policies and their implementation in the European Union on one hand and the rest of the world on the other (industrialized and developing countries alike), have resulted in disputes with serious consequences on agricultural policy, world trade and food security. A lot of research effort has been directed towards the development of marker-free transformation or systems to remove selectable markers. Such research has been in a large part motivated by perceived problems with antibiotic resistance selectable markers; however, it is not justified from a safety point of view. The aim of this review is to discuss in some detail the currently available scientific evidence that overwhelmingly argues for the safety of these marker gene systems. Our conclusion, supported by numerous studies, most of which are commissioned by some of the very parties that have taken a position against the use of antibiotic selectable marker gene systems, is that there is no scientific basis to argue against the use and presence of selectable marker genes as a class in transgenic plants.     

Three-dimensional kinematics of the human knee during walking.

Three-dimensional kinematics of the tibiofemoral joint were studied during normal walking. Target markers were fixed to tibia and femur by means of intra-cortical traction pins. Radiographs of the lower limb were obtained to compute the position of the target markers relative to internal anatomical structures. High-speed cine cameras were used to measure three-dimensional coordinates of the target markers in five subjects walking at a speed of 1.2 m s-1. Relative motion between tibia and femur was resolved according to a joint coordinate system (JCS). The measurements have identified that substantial angular and linear motions occur about and along each of the JCS axes during walking. The results do not, however, support the traditional view that the so-called 'screw home' mechanism of the knee joint operates during gait.     

QTL analysis: unreliability and bias in estimation procedures

Several statistical methods which employ multiple marker data are currently available for the analysis of quantitative trait loci (QTL) in experimental populations. Although comparable estimates of QTL location and effects have been obtained by these methods, using simulated and real data sets, their accuracy and reliability have not been extensively investigated. The present study specifically examines the merit of using F₂ and doubled haploid populations for locating QTL and estimating their effects. Factors which may affect accuracy and reliability of QTL mapping, such as the number and position of the markers available, the accuracy of the marker locations and the size of the experimental population used, are considered. These aspects are evaluated for QTL of differing heritabilities and locations along the chromosome.A population of 300 F₂ individuals and 150 doubled haploid lines gave estimates of QTL position and effect which were comparable, albeit extremely unreliable. Even for a QTL of high heritability (10%), the confidence interval was 35 cM. There was little increase in reliability to be obtained from using 300, rather than 200, F₂ individuals and 100 doubled haploid lines gave similar results to 150. QTL estimates were not significantly improved either by using the expected, rather than the observed, marker positions or by using a dense map of markers rather than a sparse map. A QTL which was asymmetrically located in the linkage group resulted in inaccurate estimates of QTL position which were seriously biassed at low heritability of the QTL. In a population of 300 F₂ individuals the bias increased from 4 cM to 20 cM, for a QTL with 10% and 2% heritability respectively.     

Mapplet: a CORBA-based genome map viewer

MOTIVATION : There are a large number of genetic and physical maps, distributed at many sites. Each site offers different kinds of access methods and viewers. CORBA, the de facto standard for distributed object-oriented computing, offers new opportunities to unify the view on these maps through standard interfaces. A collaboration of Infobiogen and the EBI proposes a common IDL for maps. RESULTS: A CORBA map viewer is presented which serves as a proof of concept for the proposed IDL. It demonstrates its usefulness in the context of map viewing and its ability to handle large maps with <1000 markers. The viewer gives access to the maps of the Radiation Hybrid Database at EBI. It gives a quick overview of several large maps side by side. The marker density at each map position is displayed and different marker types can be highlighted. AVAILABILITY: Demonstration and source code at: http://sunny.ebi.ac.uk/ approximately jungfer/Mapplet CONTACT: jungfer@ebi.ac.uk      

QTL analysis for ascochyta blight resistance in an intraspecific population of chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis> L.)

In both controlled environment and the field, six QTLs for ascochyta blight resistance were identified in three regions of the genome of an intraspecific population of chickpea using the IDS and AUDPC disease scoring systems. One QTL-region was detected from both environments, whereas the other two regions were detected from each environment. All the QTL-regions were significantly associated with ascochyta blight resistance using either of the disease scoring systems. The QTLs were verified by multiple interval mapping, and a two-QTL genetic model with considerable epistasis was established for both environments. The major QTLs generally showed additive gene action, as well as dominance inter-locus interaction in the multiple genetic model. All the QTLs were mapped near a RGA marker. The major QTLs were located on LG III, which was mapped with five different types of RGA markers. A CLRR-RGA marker and a STMS marker flanked QTL 6 for controlled environment resistance at 0.06 and 0.04 cM, respectively. Other STMS markers flanked QTL 1 for field resistance at a 5.6 cM interval. After validation, these flanking markers may be used in marker-assisted selection to breed for elite chickpea cultivars with durable resistance to ascochyta blight. The tight linkage of RGA markers to the major QTL on LG III will allow map-based cloning of the underlying resistance genes.Communicated by P. Langridge     

On Recombination between Close and Distant Markers in Phage Lambda

The contribution of parental DNA to progeny phages genetically recombinant for close markers, distant markers, or both simultaneously was studied in biparental and triparental replication-blocked crosses. The data are compatible with the previously proposed view that heterozygous overlaps at the sites of crossing over are sometimes about as long as the lambda chromosome. However, about half of the close marker recombinants have enjoyed triparental interactions, attenuating that conclusion and obscuring predictions of the long overlap model.     

Negative selection systems for transgenic barley (Hordeum vulgare L.): comparison of bacterial codA- and cytochrome P450 gene-mediated selection

Efficient negative selection systems are increasingly needed for numerous applications in plant biology. In recent years various counter-selectable genes have been tested in six dicotyledonous species, whereas there are no data available for the use of negative selection markers in monocotyledonous species. In this study, we compared the applicability and reliability of two different conditional negative selection systems in transgenic barley. The bacterial codA gene encoding cytosine deaminase, which converts the non-toxic 5-fluorocytosine (5-FC) into the toxic 5-fluorouracil (5-FU), was used for in vitro selection of germinating seedlings. Development of codA-expressing seedlings was strongly inhibited by germinating the seeds in the presence of 5-FC. For selecting plants in the greenhouse, a bacterial cytochrome P450 mono-oxygenase gene, the product of which catalyses the dealkylation of a sulfonylurea compound, R7402, into its cytotoxic metabolite, was used. T1 plants expressing the selectable marker gene showed striking morphological differences from the non-transgenic plants. In experiments with both negative selectable markers, the presence or absence of the transgene, as predicted from the physiological appearance of the plants under selection, was confirmed by PCR analysis. We demonstrate that both marker genes provide tight negative selection; however, the use of the P450 gene is more amenable to large-scale screening under greenhouse or field conditions.     

Optimal Geometrical Set for Automated Marker Placement to Virtualized Real-Time Facial Emotions

In recent years, real-time face recognition has been a major topic of interest in developing intelligent human-machine interaction systems. Over the past several decades, researchers have proposed different algorithms for facial expression recognition, but there has been little focus on detection in real-time scenarios. The present work proposes a new algorithmic method of automated marker placement used to classify six facial expressions: happiness, sadness, anger, fear, disgust, and surprise. Emotional facial expressions were captured using a webcam, while the proposed algorithm placed a set of eight virtual markers on each subject’s face. Facial feature extraction methods, including marker distance (distance between each marker to the center of the face) and change in marker distance (change in distance between the original and new marker positions), were used to extract three statistical features (mean, variance, and root mean square) from the real-time video sequence. The initial position of each marker was subjected to the optical flow algorithm for marker tracking with each emotional facial expression. Finally, the extracted statistical features were mapped into corresponding emotional facial expressions using two simple non-linear classifiers, K-nearest neighbor and probabilistic neural network. The results indicate that the proposed automated marker placement algorithm effectively placed eight virtual markers on each subject’s face and gave a maximum mean emotion classification rate of 96.94% using the probabilistic neural network.     

Variable stability of antibiotic-resistance markers in Bacillus cereus UW85 in the soybean rhizosphere in the field

We compared the stability of antibiotic-resistance markers in strains derived from Bacillus cereus UW85 in culture media and in the soybean rhizosphere in a growth chamber and in the field. We studied two independent, spontaneous mutants resistant to neomycin, three independent, spontaneous mutants resistant to streptomycin, and strains carrying plasmid pBC16, which encodes tetracycline resistance. Antibiotic-resistance markers were maintained in populations of all UW85 derivatives in culture and in the rhizosphere of soybeans grown in soil in a growth chamber. In two field experiments, antibiotic resistance was substantially lost in rhizosphere populations of B. cereus as early as 14 or as late as 116 days after planting. To distinguish between death of the inoculated strain and loss of its marker, we tested populations of B. cereus for other phenotypes (orange pigmentation, plasmid-borne resistance to tetracycline, and biocontrol activity) that are typical of UW85-derivatives used as inoculum, but atypical of the indigenous populations of B. cereus , and these phenotypes were maintained in populations from which the marker was lost. In general, neomycin-resistance markers were maintained at a higher frequency than streptomycin-resistance markers, and maintenance of antibiotic-resistance markers varied with position on the root and with the year of the experiment. In a semi-defined medium, the UW85 derivatives grew at the same rate as the wild type at 28°C, but most grew more slowly than the wild type at 16°C, demonstrating that antibiotic resistance can affect fitness under some conditions. The results suggest that the stability of antibiotic-resistance markers should be assessed in the ecosystems in which they will be studied.     

Comparative analysis of diversification and population structure of kenaf (Hibiscus cannabinus L.) and roselle (H. sabdariffa L.) using SSR and RGA (resistance gene analogue) markers

Multiple DNA marker systems and complementary analytical approaches are often useful in population genetic analysis and speciation of plants. We investigated population structure of kenaf (Hibiscus cannabinus) and roselle (H. sabdariffa) for gaining insight in evolution and geographic separation of these crop species using SSR and RGA (resistance gene analogues) markers through Bayesian clustering and principal coordinate analysis (PCoA) methods. Genotyping by 12 SSR and 16 RGA markers amplified a total of 172 loci in the study population. The RGA markers generated higher number of alleles per marker (8.2) as compared to SSR (3.4), but exhibited lower heterozygosity in the population. Genetic variance and heterozygosity in roselle population for both marker systems were lower than in kenaf. RGA markers revealed higher variation among populations. Bayesian structure as well as PCoA analysis using RGA marker revealed distinct cluster for roselle, while SSR-based classification revealed high admixture. Results indicate geographic isolation and natural selection for adaptive RGA alleles in kenaf. The Indian kenaf landraces were distinct from the exotic kenaf accessions, suggesting separate lineage formation by geographic separation. Possible origin and domestication of roselle in South India is proposed.     

Simple Tests to Detect Errors in High-Throughput Genotype Data in the Molecular Laboratory

With the advent of high-density DNA marker data sets for the mouse and other model systems, 100 or more genotypes are routinely generated from large groups of mice. Issues of the accuracy and reliability of the genotyping are extremely important but often not addressed until genetic analysis is conducted. Simple tests that rely on the robust predictions arising from Mendelian genetics can be made quickly in the molecular laboratory as the data are generated, and require only a spreadsheet program. In this report, genotype data from 392 mice tested at 96 marker sites were analyzed for errors that are typical when handling large volumes of data generated in a repetitive process. The testing consisted of: (1) repeating the genotyping of approximately 1% of the samples; (2) examining the deviation from the expected segregation ratio (1:2:1) on a marker-by-marker basis; and (3) testing the correlation of the genotype at one marker with that at neighboring genetic markers on a chromosome. These three steps allowed analysis at the level of the microtiter plate, where errors are most likely to occur. A set of 96 dinucleotide repeat markers that are polymorphic between the C57BL/6J and DBA/2J mouse strains and can be multiplexed is reported for use in other genotyping projects.     

Determination of rigid body registration marker error from edge error

Registration markers affixed to rigid bodies (fixed to bone as opposed to skin) are commonly used when tracking 3D rigid body motion. The measured positions of registration markers are subject to unavoidable errors, both systematic and non-systematic. Prior studies have investigated the error propagated to such derived properties as rigid body positions and helical axes, while others have focused on the error associated with a specific position tracking system under restricted conditions. Theoretical and simulation-based error propagation requires knowledge of the variation due to individual registration markers; however, the variation in registration marker position measurement has previously been either assumed or determined from static cases. The objective of this paper is the introduction of a method for determining individual marker variation irrespective of change in rigid body position or motion by utilizing the distances between the markers (edge lengths), which are invariant under rotation and translation. Simulations were used to validate and characterize the introduced technique, demonstrating that the predictions improve with greater edge length and additional markers, converge on reference values where the edge length is at least 4 times the magnitude of the maximum vertex variation, and that under ideal conditions the confidence interval about the predicted variation is within 7% of the maximum variation associated with that marker set. The introduced technique was tested on the results of a motion tracking experiment to demonstrate the wide disparity in vertex variation between static and non-static measurements of the same registration markers, where the non-static variation exceeded the static variation by an average factor of 12.7.     

A composite-conditional-likelihood approach for gene mapping based on linkage disequilibrium in windows of marker loci

A composite-conditional-likelihood (CCL) approach is proposed to map the position of a trait-influencing mutation (TIM) using the ancestral recombination graph (ARG) and importance sampling to reconstruct the genealogy of DNA sequences with respect to windows of marker loci and predict the linkage disequilibrium pattern observed in a sample of cases and controls. The method is designed to fine-map the location of a disease mutation, not as an association study. The CCL function proposed for the position of the TIM is a weighted product of conditional likelihood functions for windows of a given number of marker loci that encompass the TIM locus, given the sample configuration at the marker loci in those windows. A rare recessive allele is assumed for the TIM and single nucleotide polymorphisms (SNPs) are considered as markers. The method is applied to a range of simulated data sets. Not only do the CCL profiles converge more rapidly with smaller window sizes as the number of simulated histories of the sampled sequences increases, but the maximum-likelihood estimates for the position of the TIM remain as satisfactory, while requiring significantly less computing time. The simulations also suggest that non-random samples, more precisely, a non-proportional number of controls versus the number of cases, has little effect on the estimation procedure as well as sample size and marker density beyond some threshold values. Moreover, when compared with some other recent methods under the same assumptions, the CCL approach proves to be competitive.     

Tracking the motion of hidden segments using kinematic constraints and Kalman filtering

Motion capture for biomechanical applications involves in almost all cases sensors or markers that are applied to the skin of the body segments of interest. This paper deals with the problem of estimating the movement of connected skeletal segments from 3D position data of markers attached to the skin. The use of kinematic constraints has been shown previously to reduce the error in estimated segment movement that are due to skin and muscles moving with respect to the underlying segment. A kinematic constraint reduces the number of degrees of freedom between two articulating segments. Moreover, kinematic constraints can help reveal the movement of some segments when the 3D marker data otherwise are insufficient. Important cases include the human ankle complex and the phalangeal segments of the horse, where the movement of small segments is almost completely hidden from external observation by joint capsules and ligaments. This paper discusses the use of an extended Kalman filter for tracking a system of connected segments. The system is modeled using rigid segments connected by simplified joint models. The position and orientation of the mechanism are specified by a set of generalized coordinates corresponding to the mechanism's degrees of motion. The generalized coordinates together with their first time derivatives can be used as the state vector of a state space model governing the kinematics of the mechanism. The data collected are marker trajectories from skin-mounted markers, and the state vector is related to the position of the markers through a nonlinear function. The Jacobian of this function is derived. The practical use of the method is demonstrated on a model of the distal part of the limb of the horse. Monte Carlo simulations of marker data for a two-segment system connected by a joint with three degrees of freedom indicate that the proposed method gives significant improvement over a method, which does not make use of the joint constraint, but the method requires that the model is a good approximation of the true mechanism. Applying the method to data on the movement of the four distal-most segments of the horse's limb shows good between trial consistency and small differences between measured marker positions and marker positions predicted by the model.