Redistribution of Actin during Assembly and Reassembly of the Contractile Ring in Grasshopper Spermatocytes

Cytokinesis in animal cells requires the assembly of an actomyosin contractile ring to cleave the cell. The ring is highly dynamic; it assembles and disassembles during each cell cleavage, resulting in the recurrent redistribution of actin. To investigate this process in grasshopper spermatocytes, we mechanically manipulated the spindle to induce actin redistribution into ectopic contractile rings, around reassembled lateral spindles. To enhance visualization of actin, we folded the spindle at its equator to convert the remnants of the partially assembled ring into a concentrated source of actin. Filaments from the disintegrating ring aligned along reorganizing spindle microtubules, suggesting that their incorporation into the new ring was mediated by microtubules. We tracked incorporation by speckling actin filaments with Qdots and/or labeling them with Alexa 488-phalloidin. The pattern of movement implied that actin was transported along spindle microtubules, before entering the ring. By double-labeling dividing cells, we imaged actin filaments moving along microtubules near the contractile ring. Together, our findings indicate that in one mechanism of actin redistribution, actin filaments are transported along spindle microtubule tracks in a plus-end–directed fashion. After reaching the spindle midzone, the filaments could be transported laterally to the ring. Notably, actin filaments undergo a dramatic trajectory change as they enter the ring, implying the existence of a pulling force. Two other mechanisms of actin redistribution, cortical flow and de novo assembly, are also present in grasshopper, suggesting that actin converges at the nascent contractile ring from diffuse sources within the cytoplasm and cortex, mediated by spindle microtubules

Mouse mammary stem cells express prognostic markers for triple-negative breast cancer

Introduction Triple negative breast cancer (TNBC) is a heterogeneous group of tumours in which chemotherapy, the current mainstay of systemic treatment, is often initially beneficial but with a high risk of relapse and metastasis. There is currently no means of predicting which TNBC will relapse. We tested the hypothesis that the biological properties of normal stem cells are re-activated in tumour metastasis and that, therefore, the activation of normal mammary stem cell-associated gene sets in primary TNBC would be highly prognostic for relapse and metastasis. Methods Mammary basal stem and myoepithelial cells were isolated by flow cytometry and tested in low dose transplant assays. Gene expression microarrays were used to establish expression profiles of the stem and myoepithelial populations; these were compared to each other and to our previously established mammary epithelial gene expression profiles. Stem cell genes were classified by Gene Ontology (GO) analysis and the expression of a subset analysed in the stem cell population at single cell resolution. Activation of stem cell genes was interrogated across different breast cancer cohorts and within specific subtypes and tested for clinical prognostic power. Results A set of 323 genes was identified that was expressed significantly more highly in the purified basal stem cells compared to all other cells of the mammary epithelium. 109 out of 323 genes had been associated with stem cell features in at least one other study in addition to our own, providing further support for their involvement in the biology of this cell type. GO analysis demonstrated an enrichment of these genes for an association with cell migration, cytoskeletal regulation and tissue morphogenesis, consistent with a role in invasion and metastasis. Single cell resolution analysis showed that individual cells co-expressed both epithelial- and mesenchymal-associated genes/proteins. Most strikingly, we demonstrated that strong activity of this stem cell gene set in TNBCs identified those tumours most likely to rapidly progress to metastasis. Conclusions Our findings support the hypothesis that the biological properties of normal stem cells are drivers of metastasis and that these properties can be used to stratify patients with a highly heterogeneous disease such as TNBC

Development of a questionnaire specifically for patients with Ileal Orthotopic Neobladder (IONB)

The ileal orthotopic neobladder (IONB) is often used in patients undergoing radical cystectomy. The IONB allows to void avoiding the disadvantages of the external urinary diversion.In IONB patients the quality of life (QoL) appears compromised by the need to urinate voluntarily. The patients need to wake up at night interrupting the sleep-wake rhythm with consequences on social and emotional life.At present the QoL in IONB patients is evaluated by generic questionnaires. These are useful when IONB patients are compared with patients with different urinary diversions but they are less effective when only IONB patients are evaluated. To address this problem a specific questionnaire-the IONB-PRO-was developed. METHODS: A) Based on a conceptual framework, narrative-based interviews were conducted on 35 IONB patients. A basic pool of 43 items was produced and organized throughout two clinical and four QoL dimensions. An additional 15 IONB patients were interviewed for face validity testing.B) Psychometric testing was conducted on 145 IONB patients. Both classic test strategy and Rasch analysis were applied. Psychometric properties of the resulting scales were comparatively tested against other QoL-validated scales. RESULTS: The IONB-PRO questionnaire includes two sections: one on the QoL and a second section on the capability of the patient to manage the IONB. For evaluation of the QoL, three versions were delivered: 1) a basic 23-item QoL version (3 domains 23-items; alpha 0.86÷ 9.69), 2) a short-form 12-item QoL scale (alpha = 0.947), and 3) a short-form 15-item Rasch QoL scale (alpha = 0.967). Correlations of the long version scales with the corresponding dimensions of the EORTC-QLQ C30 and the EORTC-BLM30 were significant. The short forms exhibited significant correlations with the global health dimension of the EORTC-QLQ and with the urinary subscales of the EORTC-BLM30. The effect size was approximately 1.00 between patients at the 1-year follow-up period and those with 3, 5, and > 5-year follow-up periods for all scales. No relevant differences were observed between the 12-item short-form and the Rasch scale. CONCLUSIONS: The IONB-PRO long and short-forms demonstrated a high level of internal consistency and reliability with an excellent discriminanting validity

Priorities in optimisation of medical X-ray imaging - A contribution to the debate

A simplistic approach to optimising medical imaging is to use the lowest effective dose to the patient that does not jeopardise a correct diagnosis. With limited resources and over 1000 different types of X-ray examinations, it is not always easy to set the right priorities and to decide how to perform the optimisation. Recent research shows that the 'Rose model' for the detection of specific structures does not hold for realistic backgrounds. A reasonable conclusion regarding methods for optimisation is therefore not to use contrast-detail phantoms. Phantoms producing clinically realistic background images or real clinical images-modified with respect to quantum noise levels-are preferred. The images should be evaluated using visual grading or receiver operating characteristic methods. The quality of many common X-ray investigations. performed with projection techniques, is not limited by quantum noise. For these, the radiation dose to the patient can be lowered without seriously affecting the outcome of the detection task. For computed tomography (CT) investigations. the obscuring effect of anatomical structures and anatomical noise is less pronounced than in projection techniques. For (7, true optimisation in terms of a trade-off between radiation dose and image quality is therefore more likely to be effective. Both the number of CT examinations performed per year and the effective dose per examination are increasing owing to the technical advances in CT-jointly leading to a steady increase in the collective dose from CT examinations. Moreover. the smaller influence of the anatomical background in CT gives a high correlation between detection tasks and radiation dose. Thus. a reasonable view to take on which examinations to optimise is to give priority to CT examinations. The recommended distribution of a full working week for optimisation, based on the relative lifetime risk of lethal cancer from diagnostic X rays and the total collective dose from CT, is to use three out of five days to optimise CT examinations, of which one day should be devoted to paediatric CT

Method of simulating dose reduction for digital radiographic systems

The optimisation of image quality vs. radiation dose is an important task in medical imaging. To obtain maximum validity of the optimisation, it must be based on clinical images. Images at different dose levels can then either be obtained by collecting patient images at the different dose levels sought to investigate-including additional exposures and permission from an ethical committee-or by manipulating images to simulate different dose levels. The aim of the present work was to develop a method of simulating dose reduction for digital radiographic systems. The method uses information about the detective quantum efficiency and noise power spectrum at the original and simulated dose levels to create an image containing filtered noise. When added to the original image this results in an image with noise which, in terms of frequency content, agrees with the noise present in an image collected at the simulated dose level. To increase the validity, the method takes local dose variations in the original image into account. The method was tested on a computed radiography system and was shown to produce images with noise behaviour similar to that of images actually collected at the simulated dose levels. The method can, therefore, be used to modify an image collected at one dose level so that it simulates an image of the same object collected at any lower dose level

Nodule detection in digital chest radiography: Introduction to the radius chest trial

Most digital radiographic systems of today have wide latitude and are hence able to provide images with a small constraint on dose level. This opens up for an unprejudiced dose optimisation. However, in order to succeed in the optimisation task, good knowledge of the imaging and detection processes is needed. As a part of the European-wide research project 'unification of physical and clinical requirements for medical X-ray imaging'-governed by the Radiological Imaging Unification Strategies (RADIUS) Group-a major image quality trial was conducted by members of the group. The RADIUS chest trial was focused on the detection of lung nodules in digital chest radiography with the aims of determining to what extent (1) the detection of a nodule is dependent on its location, (2) the system noise disturbs the detection of lung nodules, (3) the anatomical noise disturbs the detection of lung nodules and (4) the image background and anatomical background act as pure noise for the detection of lung nodules. The purpose of the present paper is to give an introduction to the trial and describe the framework and set-up of the investigation

A software tool for increased efficiency in observer performance studies in radiology

Observer performance studies are time-consurning tasks, both for the participating observers and for the scientists collecting and analysing the data. A possible way to optimise such studies is to perform them in a completely digital environment. A software tool-ViewDEX (Viewer for Digital Evaluation of X-ray images)-has been developed in Java, enabling it to function on almost any computer. ViewDEX is designed to handle several types of studies, such as visual grading analysis (VGA), image criteria scoring (ICS) and receiver operating characteristics (ROC). The results from each observer are saved in a log file, which can be exported for further analysis in, for example, a special software for analysing ROC results. By using ViewDEX for an ROC experiment, an evaluation rate of similar to 200 images per hour can be achieved, compared to similar to 25 images per hour using hard copy evaluation. The results are obtained within minutes of completion of the viewing. The risk of human errors in the process of data collection and analysis is also minimised. The viewer has been used in a major trial containing similar to 2700 images

RADIUS--closing the circle on the assessment of imaging performance

The RADIUS (Radiological Imaging Unification Strategy) project addresses the assessment of image quality in terms of both physical and clinically relevant measures. The aim is to unify our understanding of both types of measure as well as the numerous underlying factors that play a key role in the assessments of imaging performance. In this way it is expected to provide a solid basis for the improvement in radiological safety management, where not only radiation risks are considered but also diagnostic risks of incorrect clinical outcomes (i.e. false positive/false negative). The project has applied a variety of relevant experimental and theoretical methods to this problem, which , generic to medical Imaging as a whole. Digital radiography of the chest and the breast has been employed as the clinical imaging domain vehicles for the study. The project addressed the problem from the following directions: role and relevance of pathology. human observer studies including receiver operating characteristics, image quality criteria analysis, structural noise analysis, physical measurements on clinical images, physical measurements on imaging system, modelling of imaging system, modelling of visual processes, modelling of doses delivered and IT-based scientific support strategies, This paper presents an overview of the main outcomes from this project and highlights how the research outcomes actually apply to the real world. In particular, attention will be focused on new and original findings and methods and techniques that have been developed within the framework of the project. The relevance of the project's outcomes to future European research will also be presented

Demonstration of correlations between clinical and physical image quality measures in chest and lumbar spine screen-film radiography

The ability to predict clinical image quality from physical measures is useful for optimization in diagnostic radiology. In this work, clinical and physical assessments of image quality are compared and correlations between the two are derived. Clinical assessment has been made by a group of expert radiologists who evaluated fulfilment of the European image criteria for chest and lumbar spine radiography using two scoring methods: image criteria score (ICS) and visual grading analysis score (VGAS). Physical image quality measures were calculated using a Monte Carlo simulation model of the complete imaging system. This model includes a voxelized male anatomy and was used to calculate contrast and signal-to-noise ratio of various Important anatomical details and measures of dynamic range. Correlations between the physical image quality measures on the one hand and the ICS and VGAS on the other were sought. 16 chest and 4 lumbar spine imaging system configurations were compared in frontal projection. A statistically significant correlation with clinical image quality was found in chest posteroanterior radiography for the contrast of blood vessels in the retrocardiac area and a measure of useful dynamic range. In lumbar spine anteroposterior radiography, a similar significant correlation with clinical image quality was found between the contrast and signal-to-noise ratio of the trabecular structures in the L1-L5 vertebrae. The significant correlation shows that clinical image duality can, at least in some cases, be predicted from appropriate measures of physical image quality

Imaging of root canal fillings: a comparison of subjective image quality between limited cone-beam CT, storage phosphor and film radiography

WOS: 000244229100003PubMed ID: 17305694Aim To compare the subjective quality of limited cone-beam computed tomography (LCBCT), storage phosphor plate (SPP) and F-speed film images for the evaluation of length and homogeneity of root fillings. Methodology Root canals of 17 extracted permanent mandibular incisor teeth were filled. With the teeth placed in their jaws, images were obtained with Accu-I-Tomo LCBCT, Digora((R)) Optime image plate system and F-speed film using exposure parameters yielding 'clinically' acceptable density and contrast. Three radiologists and three endodontists independently rated the quality of all images in respect to homogeneity and the length of root fillings using a 3-graded scale. Evaluations were undertaken in two sessions. In the first, the coronal LCBCT images were not included. In the second, both coronal and sagittal LCBCT images were rated along with F-speed film and SPP images. Results were compared using the Friedman test (P < 0.05). Pair-wise comparisons of systems were completed using the Wilxocon signed-ranks test (P < 0.05). Kappa was used to measure interobserver agreement. Results Digora images were rated superior, consecutively followed by F-speed films and LCBCT images, for the evaluation of both homogeneity and length of root fillings in both the evaluation sessions (P < 0.05). Kappa ranged from slight to moderate for the length evaluation of root fillings and from poor to fair for the evaluation of homogeneity of root fillings. Conclusion Image quality of storage phosphor images was subjectively as good as conventional film images and superior to LCBCT images for the evaluation of both homogeneity and length of root fillings in single-rooted teeth