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Is morphogenesis an intrinsically robust process?

Because morphogenesis is dynamically complex, involving many processes operating over different spatial and temporal scales, it is generally assumed that the stability and repeatability of development depends primarily upon accurate control over parameter values i.e. a precise genetic program. However, it is possible that the dynamic coupling between the different mechanisms involved in development reduces the choices available to the system because of bias in successive symmetry-breaking events. Evidence for such stable cascades in morphogenetic processes is described, and a general argument is presented for morphogenesis as an intrinsically robust process. The evolutionary implications of the hypothesis are considerable.

Acetabularia↗

Robust oscillations within the interlocked feedback model of Drosophila circadian rhythm.

A mechanism for generating circadian rhythms has been of major interest in recent years. After the discovery of per and tim, a model with a simple feedback loop involving per and tim has been proposed. However, it is recognized that the simple feedback model cannot account for phenotypes generated by various mutants. A recent report by Glossop, Lyons & Hardin [Science286, 766 (1999)] on Drosophila suggests involvement of another feedback loop by dClk that is interlocked with per-tim feedback loop. In order to examine whether interlocked feedback loops can be a basic mechanism for circadian rhythms, a mathematical model was created and examined. Through extensive simulation and mathematical analysis, it was revealed that the interlocked feedback model accounts for the observations that are not explained by the simple feedback model. Moreover, the interlocked feedback model has robust properties in oscillations.

Animals↗

Theory of robustness of irreversible differentiation in a stem cell system: chaos hypothesis.

Based on an extensive study of a dynamical systems model of the development of a cell society, a novel theory for stem cell differentiation and its regulation is proposed as the "chaos hypothesis". Two fundamental features of stem cell systems-stochastic differentiation of stem cells and the robustness of a system due to regulation of this differentiation-are found to be general properties of a system of interacting cells exhibiting chaotic intra-cellular reaction dynamics and cell division, whose presence does not depend on the detail of the model. It is found that stem cells differentiate into other cell types stochastically due to a dynamical instability caused by cell-cell interactions, in a manner described by the Isologous Diversification theory. This developmental process is shown to be stable not only with respect to molecular fluctuations but also with respect to the removal of cells. With this developmental process, the irreversible loss of multipotency accompanying the change from a stem cell to a differentiated cell is shown to be characterized by a decrease in the chemical diversity in the cell and of the complexity of the cellular dynamics. The relationship between the division speed and the loss of multipotency is also discussed. Using our model, some predictions that can be tested experimentally are made for a stem cell system.

Animals↗

Robustness of the estimator of the index of dispersion for DNA sequences.

If substitutions in DNA sequences follow a Poisson process, the ratio of the variance in the number of substitutions to the mean number of substitutions (the index of dispersion) should equal 1. In this paper, the robustness of the commonly applied estimator of the index of dispersion in replacement sites and silent sites to various assumptions regarding DNA evolution is explored using simulation methods. The estimate of the index of dispersion may be strongly biased if the assumptions of the model of substitution are violated. However, the results of this study support the conclusions of studies by Gillespie and Ohta that the process of substitution in replacement sites is overdispersed. This result contradicts those of a recent study and shows that the high index of dispersion for replacement sites is not an artifact caused by the method of estimation.

Computer Simulation↗

Robust and efficient regulation of transgene expression in vivo by improved tetracycline-dependent lentiviral vectors.

We developed a panel of lentiviral vectors that displayed tetracycline-regulated transgene expression over two orders of magnitude in bulk, non-selected populations of transduced cells in vitro and in vivo. The robust expression and homogeneous response indicated that most transduced vector genomes were transcription competent and responsive to regulation, providing the lentiviral vector with a novel competitive advantage for gene transfer. After ex vivo transduction and transplantation of cord blood CD34+ cells into NOD/SCID mice, reporter gene expression could be switched "on" and "off" in human hematopoietic cells in vivo for prolonged times, proving integration of the regulated expression system into long-term repopulating cells. By vector injection into established tumor grafts, we achieved efficient delivery and quantitative regulation of transgene expression in vivo. By these approaches, gene function studies can now be performed in in vivo models of human hematopoiesis and cancer. In the future, regulated lentiviral vectors will improve the safety and efficacy of gene therapy.

Animals↗

New robust 3-D phase unwrapping algorithms: application to magnetic field mapping and undistorting echoplanar images.

The phase, as well as the magnitude, of MRI images can carry useful information. It may be used to encode flow or temperature, or to map the magnetic field for the undistorting of EPIs and automated shimming. In all cases, we measure the extra spin given to nuclei. Unfortunately, we can only measure the final phase of the spins: the rotation is wrapped into the range [-pi, +pi], and to obtain a measure of the parameter of interest the missing multiples of 2pi must be replaced--a process known as phase unwrapping. While simple in principle, standard phase unwrapping algorithms fail catastrophically in the presence of even small amounts of noise. Here we present a new algorithm for robust three-dimensional phase unwrapping, in which unwrapping is guided, so that it initially works on less noisy regions. We test the algorithm on simulated phase data, and on maps of magnetic field, which were then used to successfully undistort EPI images. The unwrapping algorithm could be directly applied to other kinds of phase data.

Acoustic Stimulation↗

A robust method for extraction and automatic segmentation of brain images.

A new protocol is introduced for brain extraction and automatic tissue segmentation of MR images. For the brain extraction algorithm, proton density and T2-weighted images are used to generate a brain mask encompassing the full intracranial cavity. Segmentation of brain tissues into gray matter (GM), white matter (WM), and cerebral spinal fluid (CSF) is accomplished on a T1-weighted image after applying the brain mask. The fully automatic segmentation algorithm is histogram-based and uses the Expectation Maximization algorithm to model a four-Gaussian mixture for both global and local histograms. The means of the local Gaussians for GM, WM, and CSF are used to set local thresholds for tissue classification. Reproducibility of the extraction procedure was excellent, with average variation in intracranial capacity (TIC) of 0.13 and 0.66% TIC in 12 healthy normal and 33 Alzheimer brains, respectively. Repeatability of the segmentation algorithm, tested on healthy normal images, indicated scan-rescan differences in global tissue volumes of less than 0.30% TIC. Reproducibility at the regional level was established by comparing segmentation results within the 12 major Talairach subdivisions. Accuracy of the algorithm was tested on a digital brain phantom, and errors were less than 1% of the phantom volume. Maximal Type I and Type II classification errors were low, ranging between 2.2 and 4.3% of phantom volume. The algorithm was also insensitive to variation in parameter initialization values. The protocol is robust, fast, and its success in segmenting normal as well as diseased brains makes it an attractive clinical application.

Adult↗

Classification of structural images via high-dimensional image warping, robust feature extraction, and SVM.

This paper presents a method for classification of medical images, using machine learning and deformation-based morphometry. A morphological representation of the anatomy of interest is first obtained using high-dimensional template warping, from which regions that display strong correlations between morphological measurements and the classification (clinical) variable are extracted using a watershed segmentation, taking into account the regional smoothness of the correlation map which is estimated by a cross-validation strategy in order to achieve robustness to outliers. A Support Vector Machine-Recursive Feature Elimination (SVM-RFE) technique is then used to rank computed features from the extracted regions, according to their effect on the leave-one-out error bound. Finally, SVM classification is applied using the best set of features, and it is tested using leave-one-out. The results from a group of 61 brain images of female normal controls and schizophrenia patients demonstrate not only high classification accuracy (91.8%) and steep ROC curves, but also exceptional stability with respect to the number of selected features and the SVM kernel size.

Algorithms↗

Robust visualization of the dental occlusion by a double scan procedure.

A detailed visualization of the dental occlusion in 3D image-based planning environments for oral and maxillofacial planning is important. With CT imaging however, this occlusion is often deteriorated by streak artifacts caused by amalgam fillings. Moreover, more detailed surface information at the level of the dental cuspids is often desired. In this paper, a double scan technique is introduced to image the dental occlusion by means of a newly designed 3D splint. The patient wears this splint between the upper and lower teeth during CT-scan. In a second step, the splint is positioned between the plaster casts of the upper and lower jaw, and this setup is scanned. Based on markers in the 3D splint, both data sets are fused and a combined visualization is possible. The accuracy, robustness and applicability in clinical routine is shown. This technology enables meticulous 3D cephalometric analysis, detailed maxillofacial planning and opens possibilities towards intraoperative support.

Cadaver↗

STREM: a robust multidimensional parametric method to segment MS lesions in MRI.

We propose to segment Multiple Sclerosis (MS) lesions overtime in multidimensional Magnetic Resonance (MR) sequences. We use a robust algorithm that allows the segmentation of the abnormalities using the whole time series simultaneously and we propose an original rejection scheme for outliers. We validate our method using the BrainWeb simulator. To conclude, promising preliminary results on longitudinal multi-sequences of clinical data are shown.

Algorithms↗

A robust orthogonal algorithm for system identification and time-series analysis.

We describe and illustrate methods for obtaining a parsimonious sinusoidal series representation or model of biological time-series data. The methods are also used to identify nonlinear systems with unknown structure. A key aspect is a rapid search for significant terms to include in the model for the system or the time-series. For example, the methods use fast and robust orthogonal searches for significant frequencies in the time-series, and differ from conventional Fourier series analysis in several important respects. In particular, the frequencies in our resulting sinusoidal series need not be commensurate, nor integral multiples of the fundamental frequency corresponding to the record length. Freed of these restrictions, the methods produce a more economical sinusoidal series representation (than a Fourier series), finding the most significant frequencies first, and automatically determine model order. The methods are also capable of higher resolution than a conventional Fourier series analysis. In addition, the methods can cope with unequally-spaced or missing data, and are applicable to time-series corrupted by noise. Finally, we compare one of our methods with a well-known technique for resolving sinusoidal signals in noise using published data for the test time-series.

Algorithms↗

Robustness of a model of exercise.

A recent longitudinal study of exercise behavior was published in this journal. A structural equation model derived from the theory of reasoned action was evaluated and found to fit the data well. This paper presents further analysis of the data to determine the robustness of the results to modifications of the original model.

Computer Simulation↗

A robust assessment of the NoGo-anteriorisation of P300 microstates in a cued Continuous Performance Test.

The Continuous Performance Test (CPT) is successfully applied in clinical routine to evaluate attentional performance. The aim of the present study was to investigate the features of the ERPs related to the conditions of a cued CPT, in particular the Go- and the NoGo-condition demanding either the execution or the inhibition of a prepared motor response. For that purpose, 21-channel-ERPs of ten healthy subjects elicited by the Go, NoGo, primer and distractor cues were analyzed with reference-independent methods. The P300 microstates were identified by means of a data-driven segmentation of the ERPs based on the individual peaks of the Global Field Power (GFP). The topographical assessment of the P300 fields yielded an extraordinarily robust result consisting of a more anterior location of the positive centroid in the NoGo compared to the Go condition in every single subject. In conclusion, this result is an impressive validation of the applied reference-independent spatial analysis which reveals the rapid changes of the brain electrical field configurations related to the execution/inhibition paradigm within the cued CPT. Because of the stability of the NoGo anteriorisation we propose to use this parameter as a topographical standard index, analogous to the amplitude effect between oddball targets and nontargets which defines the classical P300.

Adult↗

Robust design of finger probe in non-invasive total haemoglobin monitor.

The development of a non-invasive monitor of total haemoglobin has been reported previously. The paper presents the design and testing of a finger probe used in that optical haemoglobin monitor. When a finger is inserted into the probe, light is radiated onto the nail, and a silicon detector measures transmitted light. This finger probe can have different values or settings for design parameters such as the internal colour, detector area, the emission area of a light source and the distance between the light source and detector. Design of experiment (DOE) was introduced to select the best combination of design parameters that were robust to external conditions such as finger alignment and ambient light. An optimally designed finger probe from DOE analysis, compared with the initial design, increased the correlation coefficient from 0.696 to 0.869 and improved the standard deviation from 1.18 to 0.81 g dl(-1) in predicting total haemoglobin. This was under different conditions of finger-probe alignment. Under different ambient light conditions, the optimum design improved the correlation coefficient from 0.735 to 0.870 and reduced the standard deviation from 1.14 to 0.83 g dl(-1).

Equipment Design↗

Real-time system for robust spectral parameter estimation in Doppler signal analysis.

In assessing the level of stenosis in extracranial Doppler analysis, spectral analysis has until now been used qualitatively, for the most part. Owing to the many variables affecting the measurements (mainly noise level and instrument setting made subjectively by the operator), the reliability of the inferences on the degree of stenosis is not clearly definable. Under such conditions the need arises for algorithms and systems that can estimate spectral parameters with a higher degree of accuracy, to verify whether reliable inferences can indeed by made or if this technique is only a qualitative one. In the paper a real-time spectral analysis system is described. The system relies on a new spectral estimation algorithm which gives estimates with good robustness with respect to noise. Moreover, a clear measurement procedure which eliminates the many subjective factors affecting the estimates has also been proposed and used. The system has been evaluated with simulated signals and in clinical trials and has shown better performance than the commonly used commercial analysers.

Adult↗

Hierarchically coupled ultradian oscillators generating robust circadian rhythms.

Ensembles of mutually coupled ultradian cellular oscillators have been proposed by a number of authors to explain the generation of circadian rhythms in mammals. Most mathematical models using many coupled oscillators predict that the output period should vary as the square root of the number of participating units, thus being inconsistent with the well-established experimental result that ablation of substantial parts of the suprachiasmatic nuclei (SCN), the main circadian pacemaker in mammals, does not eliminate the overt circadian functions, which show no changes in the phases or periods of the rhythms. From these observations, we have developed a theoretical model that exhibits the robustness of the circadian clock to changes in the number of cells in the SCN, and that is readily adaptable to include the successful features of other known models of circadian regulation, such as the phase response curves and light resetting of the phase.

Activity Cycles↗

Identification of physiological systems: a robust method for non-parametric impulse response estimation.

The identification of non-parametric impulse response functions (IRFs) from noisy finite-length data records is analysed using the techniques of matrix perturbation theory. Based on these findings, a method for IRF estimation is developed that is more robust than existing techniques, particularly when the input is non-white. Furthermore, methods are developed for computing confidence bounds on the resulting IRF estimates. Monte Carlo simulations are used to assess the capabilities of this new method and to demonstrate its superiority over classical techniques. An application to the identification of dynamic ankle stiffness in humans is presented.

Ankle Joint↗

Robust ability of IFN-gamma to upregulate class II MHC antigen expression in tumor bearing rat brains.

T cells are attractive for delivering therapy to brain tumor, especially disseminated micro-tumor. However, to trigger effector function, tumor antigen must be re-presented to T cells, via major histocompatibility complex (MHC) proteins, at the tumor site. In normal brain, MHC+ antigen-presenting cells (APC) are rare, but abundant after gamma interferon (IFN-gamma) injection. Here we studied tumor-bearing brains. IFN-gamma (or buffer) was injected stereotactically into brains with established tumors from a panel of immunologically varied glioma cell lines, some expressing b-galactosidase as a micro-tumor marker. Four days later, cryostat sections were stained for tumor and MHC proteins. In phosphate-buffered saline-injected controls, class II MHC+ potential APC (microglia, macrophages) were seen only at (some) tumor sites. In rats that received IFN-gamma, class II+ potential APC were widespread, including all actual and potential micro-tumor sites and all tumor-free areas. In the same slides, neither class I nor class II MHC antigen was detected in neural cells or most tumor cells. This MHC pattern favors indirect re-presentation of tumor antigen, by tumor-adjacent APC. The robust response to IFN-gamma might also be exploited in other ways: activated microglia and macrophages can attack tumor directly, and class II+ APC may help mark micro-tumor sites.

Animals↗