Automatic neutrophil nucleus lobe counting based on graph representation of region skeleton

Abnormal neutrophil nucleus lobation (like left shift and right shift) helps to diagnose for some clinical conditions. Currently, quantification of it depends on the manual microscopic inspection of blood smears by clinicians. The quality of the manual inspection is extremely limited by the efficiency of clinicians and their medical background. This article proposed an automatic lobe counting method based on the graph representation of the nucleus region skeletons. Skeletons of the segmented nucleus regions are computed by augmented Fast Marching Method and transformed into plane graphs. Then the nucleus lobes are split based on the extracted graph properties including width distribution along the skeleton and graph structure decomposition. Experiments show that the proposed method could efficiently approaches the real lobe numbers in blood smears and reliably distinguish the stabs from the segmented neutrophils, thus it should be helpful in automatic neutrophil lobe number quantification and abnormal lobation diagnosis. ? 2012 International Society for Advancement of Cytometry.     

Regeneration of a well-differentiated human airway surface epithelium by spheroid and lentivirus vector-transduced airway cells

BACKGROUND: Following injury to the airway epithelium, rapid regeneration of a functional epithelium is necessary in order to restore the epithelial barrier integrity. In the perspective of airway gene/cell therapy, we analyzed the capacity of human airway epithelial cells cultured as three-dimensional (3-D) spheroid structures to be efficiently transduced on long term by a pseudotyped lentiviral vector. The capacity of the 3-D spheroid structures to repopulate a denuded tracheal basement membrane and regenerate a well-differentiated airway epithelium was also analyzed. METHODS: An HIV-1-derived VSV-G pseudotyped lentiviral vector encoding the enhanced green fluorescent protein (eGFP) was used. Airway epithelial cells were isolated from mature human fetal tracheas and airway xenografts, cultured as 3-D spheroid structures, and either transduced at multiplicity of infection (MOI) 10 and 100 or assayed in an ex vivo and in vivo model to evaluate their regeneration capacity. RESULTS: An in vivo repopulation assay in SCID-hu mice with transduced isolated fetal airway epithelial cells shows that lentiviral transduction does not alter the airway reconstitution. Transduction of the 3-D spheroid structures shows that 12% of cells were eGFP-positive for up to 80 days. In ex vivo and in vivo assays (NUDE-hu mice), the 3-D spheroid structures are able to repopulate denuded basement membrane and reconstitute a well-differentiated human airway surface epithelium. CONCLUSIONS: The efficient and long-term lentiviral transduction of 3-D spheroid structures together with their capacity to regenerate a well-differentiated mucociliary epithelium demonstrate the potential relevance of these 3-D structures in human airway gene/cell therapy.     

Development and validation of a subject-specific finite element model of a human clavicle

This study aimed to develop and validate a finite element (FE) model of a human clavicle which can predict the structural response and bone fractures under both axial compression and anterior–posterior three-point bending loads. Quasi-static non-injurious axial compression and three-point bending tests were first conducted on a male clavicle followed by a dynamic three-point bending test to fracture. Then, two types of FE models of the clavicle were developed using bone material properties which were set to vary with the computed tomography image density of the bone. A volumetric solid FE model comprised solely of hexahedral elements was first developed. A solid-shell FE model was then created which modelled the trabecular bone as hexahedral elements and the cortical bone as quadrilateral shell elements. Finally, simulations were carried out using these models to evaluate the influence of variations in cortical thickness, mesh density, bone material properties and modelling approach on the biomechanical responses of the clavicle, compared with experimental data. The FE results indicate that the inclusion of density-based bone material properties can provide a more accurate reproduction of the force–displacement response and bone fracture timing than a model with uniform bone material properties. Inclusion of a variable cortical thickness distribution also slightly improves the ability of the model to predict the experimental response. The methods developed in this study will be useful for creating subject-specific FE models to better understand the biomechanics and injury mechanism of the clavicle.     

Automatic localization of anatomical landmarks on the back surface and construction of a body-fixed coordinate system

A method for automatic measurement of anatomical landmarks on the back surface is presented. The landmarks correspond to the verteba prominens, the dimples of the posterior superior iliac spines and the sacrum point (beginning of rima ani), which are characterized by distinct surface curvature. The surface curvatures are calculated from rasterstereographic surface measurements. The procedure of isolating a region of interest for each landmark (surface segmentation) and the calculation of the landmark coordinates are described in detail. The accuracy of landmark localization was tested with serial rasterstereographs of 28 patients (with moderate idiopathic scoliosis). From the results the intrinsic accuracy of the method is estimated to be little more than 1 mm (depending on the sampling density of the surface measurement). Therefore, the landmarks may well be used for the objective definition of a body-fixed reference coordinate system. The accuracy is, however, dependent on the specific landmark and a minor influence of posture variations is observed.     

An articular cartilage contact model based on real surface geometry

Abnormal, excessive stresses acting on articular joint surfaces are speculated to be one of the causes for joint degeneration. However, articular surface stresses have not been studied systematically, since it is technically difficult to measure in vivo contact areas and pressures in dynamic situations. Therefore, we implemented a numerical model of articular surface contact using accurate surface geometries. The model was developed for the cat patellofemoral joint. We demonstrated that small misalignments of the patella relative to the femur change the joint contact mechanics substantially for a given external load. These results suggest that misalignment might be studied as one of the factors causing articular cartilage disorder and joint degeneration.     

A parametric geometry model of the aortic valve for subject-specific blood flow simulations using a resistive approach

Cardiac valves simulation is one of the most complex tasks in cardiovascular modeling. Fluid–structure interaction is not only highly computationally demanding but also requires knowledge of the mechanical properties of the tissue. Therefore, an alternative is to include valves as resistive flow obstacles, prescribing the geometry (and its possible changes) in a simple way, but, at the same time, with a geometry complex enough to reproduce both healthy and pathological configurations. In this work, we present a generalized parametric model of the aortic valve to obtain patient-specific geometries that can be included into blood flow simulations using a resistive immersed implicit surface (RIIS) approach. Numerical tests are presented for geometry generation and flow simulations in aortic stenosis patients whose parameters are extracted from ECG-gated CT images.

     

Generation of hemipelvis surface geometry based on statistical shape modelling and contralateral mirroring

Biomechanics and Modeling in Mechanobiology - Personalised fracture plates manufactured using 3D printing offer an improved treatment option for unstable pelvic ring fractures that may not be...     

Effect of position and lung volume on upper airway geometry

The occurrence of upper airway obstruction during sleep and with anesthesia suggests the possibility that upper airway size might be compromised by the gravitational effects of the supine position. We used an acoustic reflection technique to image airway geometry and made 180 estimates of effective cross-sectional area as a function of distance along the airway in 10 healthy volunteers while they were supine and also while they were seated upright. We calculated z-scores along the airway and found that pharyngeal cross-sectional area was smaller in the supine than in the upright position in 9 of the 10 subjects. For all subjects, pharyngeal cross-sectional area was 23 +/- 8% smaller in the supine than in the upright position (P less than or equal to 0.05), whereas glottic and tracheal areas were not significantly altered. Because changing from the upright to the supine position causes a decrease in functional residual capacity (FRC), six of these subjects were placed in an Emerson cuirass, which was evacuated producing a positive transrespiratory pressure so as to restore end-expiratory lung volume to that seen before the position change. In the supine posture an increase in end-expiratory lung volume did not change the cross-sectional area at any point along the airway. We conclude that pharyngeal cross-sectional area decreases as a result of a change from the upright to the supine position and that the mechanism of this change is independent of the change in FRC.     

Automatic detection of surface EMG activation timing using a wavelet transform based method

The problem of the identification of the muscle contraction timing by using surface electromyographic signal is addressed. The timing detection of the muscular activation in dynamic conditions has a real clinical diagnostic impact. Widely used single threshold methods still rely on the experience of the operator in manually setting that threshold. A new approach to detect the muscular activation intervals, that is based on discontinuities detection in the wavelet domain, is proposed. Accuracy and precision of the algorithm were assessed by using a set of simulated signals obtaining values lower than 11.0 and 8.7 ms for biases and standard deviations of the estimation, respectively. Moreover an experimental application of the algorithm was carried out recruiting a population of 10 able-bodied subjects and processing the myoelectric signals recorded from the lower limb during an isokinetic exercise. The algorithm was able to reveal correctly the timing of muscular activation with performance comparable to the state-of-the-art methods. The detection algorithm is automatic and user-independent, it manages the detection of both onset and offset activation, it can be fruitfully applied even in presence of noise and, therefore, it can be used also by unskilled operators.     

Cross sectional geometry of the forelimb skeleton and flight mode in pelecaniform birds

Avian wing elements have been shown to experience both dorsoventral bending and torsional loads during flapping flight. However, not all birds use continuous flapping as a primary flight strategy. The pelecaniforms exhibit extraordinary diversity in flight mode, utilizing flapping, flap‐gliding, and soaring. Here we (1) characterize the cross‐sectional geometry of the three main wing bone (humerus, ulna, carpometacarpus), (2) use elements of beam theory to estimate resistance to loading, and (3) examine patterns of variation in hypothesized loading resistance relative to flight and diving mode in 16 species of pelecaniform birds. Patterns emerge that are common to all species, as well as some characteristics that are flight‐ and diving‐mode specific. In all birds examined, the distal most wing segment (carpometacarpus) is the most elliptical (relatively high I_max/I_min) at mid‐shaft, suggesting a shape optimized to resist bending loads in a dorsoventral direction. As primary flight feathers attach at an oblique angle relative to the long axis of the carpometacarpus, they are likely responsible for inducing bending of this element during flight. Moreover, among flight modes examined the flapping group (cormorants) exhibits more elliptical humeri and carpometacarpi than other flight modes, perhaps pertaining to the higher frequency of bending loads in these elements. The soaring birds (pelicans and gannets) exhibit wing elements with near‐circular cross‐sections and higher polar moments of area than in the flap and flap‐gliding birds, suggesting shapes optimized to offer increased resistance to torsional loads. This analysis of cross‐sectional geometry has enhanced our interpretation of how the wing elements are being loaded and ultimately how they are being used during normal activities. J. Morphol., 2011. © 2011 Wiley‐Liss,Inc.     

3,3-diphenylpentane skeleton as a steroid skeleton substitute: novel inhibitors of human 5alpha-reductase 1

We designed and synthesized novel type 1 5alpha-reductase inhibitors by using 3,3-diphenylpentane skeleton as a substitute for the usual steroid skeleton. 4-(3-(4-(N-Methylacetamido)phenyl)pentan-3-yl)phenyl dibenzylcarbamate (11k) is a competitive 5alpha-reductase inhibitor with the IC(50) value of 0.84 microM.     

Effects of surface tension and viscosity on airway reopening

We studied airway opening in a benchtop model intended to mimic bronchial walls held in apposition by airway lining fluid. We measured the relationship between the airway opening velocity (U) and the applied airway opening pressure in thin-walled polyethylene tubes of different radii (R) using lining fluids of different surface tensions (gamma) and viscosities (mu). Axial wall tension (T) was applied to modify the apparent wall compliance characteristics, and the lining film thickness (H) was varied. Increasing mu or gamma or decreasing R or T led to an increase in the airway opening pressures. The effect of H depended on T: when T was small, opening pressures increased slightly as H was decreased; when T was large, opening pressure was independent of H. Using dimensional analysis, we found that the relative importance of viscous and surface tension forces depends on the capillary number (Ca = microU/gamma). When Ca is small, the opening pressure is approximately 8 gamma/R and acts as an apparent "yield pressure" that must be exceeded before airway opening can begin. When Ca is large (Ca greater than 0.5), viscous forces add appreciably to the overall opening pressures. Based on these results, predictions of airway opening times suggest that airway closure can persist through a considerable portion of inspiration when lining fluid viscosity or surface tension are elevated.     

Measles virus preferentially transduces the basolateral surface of well-differentiated human airway epithelia

Measles virus (MV) is typically spread by aerosol droplets and enters via the respiratory tract. The progression of MV infection has been widely studied; yet, the pathway for virus entry in polarized human airway epithelia has not been investigated. Herein we report the use of a replication-competent Edmonston vaccine strain of MV expressing enhanced green fluorescent protein (MV-eGFP) to infect primary cultures of well-differentiated human airway epithelia. Previous studies with polarized Caco-2 cells (intestine-derived human epithelia) and MDCK cells (kidney-derived canine epithelia) demonstrated that MV primarily infected and exited the apical surface. In striking contrast, our results indicate that MV preferentially transduces human airway cells from the basolateral surface; however, virus release remains in an apical direction. When MV-eGFP was applied apically or basolaterally to primary cultures of airway epithelia, discrete foci of eGFP expression appeared and grew; however, the cell layer integrity was maintained for the duration of the study (7 days). Interestingly, utilizing immunohistochemistry and confocal microscopy, we observed widespread expression of the receptor for the vaccine strain of MV (CD46) at greatest abundance on the apical surface of the differentiated human airway epithelia as well as in human tracheal tissue sections. These data suggest that the progression of MV infection through the respiratory epithelium may involve pathways other than direct binding and entry through the apical surface of airway epithelia.     

Erythromycin increases bactericidal activity of surface liquid in human airway epithelial cells

Macrolide antibiotics have clinical benefits in patients with diffuse panbronchiolitis and in patients with cystic fibrosis. Although many mechanisms have been proposed, the precise mechanisms are still uncertain. We examined the effects of erythromycin on bactericidal activity of airway surface liquid secreted by cultured human tracheal epithelial cells. Airway surface liquid was collected by washing the surface of human tracheal epithelial cells with a sodium solution (40 meq/l). Methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa were incubated with airway surface liquid, and the number of surviving bacteria was examined. The number of bacteria in airway surface liquid from the cells cultured in medium alone was significantly lower than that in the sodium solution. Furthermore, the number of bacteria in airway surface liquid from the cells treated with erythromycin was significantly lower than that in airway surface liquid from the cells treated with solvent alone. The production of mRNA and protein of human beta-defensin-1 and human beta-defensin-2 was significantly increased by erythromycin. Bactericidal activity of airway surface liquid was observed at low concentrations (40 meq/l) of sodium but not at higher concentrations (> or =80 meq/l). Airway surface liquid did not contain significant amounts of antibiotics supplemented in the culture medium. Erythromycin at the levels in airway surface liquid and in culture medium did not inhibit bacterial growth. These results suggest that erythromycin may increase bactericidal activity of airway surface liquid in human airway epithelial cells through human beta-defensins production and reduce susceptibility of the airway to bacterial infection.     

Synergistic and additive killing by antimicrobial factors found in human airway surface liquid

Airway surface liquid contains multiple factors thought to provide a first line of defense against bacteria deposited in the airways. Although the antimicrobial action of individual factors has been studied, less is known about how they work in combination. We examined the combined action of six antimicrobial peptides found in airway surface liquid. The paired combinations of lysozyme-lactoferrin, lysozyme-secretory leukocyte protease inhibitor (SLPI), and lactoferrin-SLPI were synergistic. The triple combination of lysozyme, lactoferrin, and SLPI showed even greater synergy. Other combinations involving the human beta-defensins, LL-37, and tobramycin (often administered to cystic fibrosis patients by inhalation) were additive. Because the airway surface liquid salt concentration may be elevated in cystic fibrosis patients, we examined the effect of salt on the synergistic combinations. As the ionic strength increased, synergistic interactions were lost. Our data suggest that the antibacterial potency of airway surface liquid may be significantly increased by synergistic and additive interactions between antimicrobial factors. These results also suggest that increased salt concentrations that may exist in cystic fibrosis could inhibit airway defenses by diminishing these synergistic interactions.     

Proteomic analysis of a membrane skeleton fraction from human liver

The cytoskeleton networks around liver cell cortex can resist Triton extraction and co-pellet with their tightly associated integral membrane proteins, forming assemblies called "membrane skeletons". Despite their important roles in determining cell shape and in signal transduction pathways, the membrane skeletons of human liver cells are uncharacterized to a great extent. In the present work, we prepared a membrane skeleton fraction by Triton extraction of human liver plasma membranes and then separated its protein components by 2-D gels. We optimized the detergent used for protein solubilization and found that 2% ASB-14 allowed the best recovery of membrane skeleton proteins. By analyzing the protein spots with MALDI-TOF and MALDI-TOF-TOF MS, we identified 104 nonredundant proteins, wherein 38 were cytoskeletal proteins that were further classified into several groups, including proteins in fodrin-based meshworks, adhesion proteins (proteins involved in adherens junctions, focal adhesions, desmosomes, hemidesmosomes and tight junctions), proteins that regulate F-actin dynamics, motor proteins, and some other cytoskeletal proteins. To the best of our knowledge, this is one of the largest data sets of membrane skeleton proteins to date. All the results suggested that the liver cells had complex actin- and cytokeratin-based membrane skeletons. This work provided a representative 2-DE map of membrane skeletons from human normal liver, for the purpose of helping to elucidate the composition and function of the membrane skeletons.     

Three‐dimensional geometry of a pterosaur wing skeleton,and its implications for aerial and terrestrial locomotion

This study reports on the three‐dimensional spatial arrangement and movements of the skeleton of Anhanguera santanae (Pterodactyloidea: Ornithocheiridae), determined using exceptionally well‐preserved uncrushed fossil material, and a rigid‐body method for analysing the joints of extinct animals. The geometric results of this analysis suggest that the ornithocheirids were inherently unstable in pitch and yaw. As a result, pitch control would probably have been brought about by direct adjustment of the angle of attack of the wing, by raising or lowering the trailing edge from the root using the legs if, as is indicated in soft‐tissue specimens of a number of unrelated pterosaur species, the legs were attached to the main wing membrane, or by using long‐axis rotations at the shoulder or wrist to raise and lower the trailing edge from the wingtip. An analysis of the three‐dimensional morphology of the wrist lends support to the idea that the pteroid – a long, slender wrist bone unique to pterosaurs that supported a membranous forewing – was directed forwards in flight, not towards the body. As a result, the forewing could have fulfilled the function of an air‐brake and high‐lift device, and may also have had an important role in pitch, yaw, and roll control. The joint analysis is consistent with a semi‐erect quadrupedal model of terrestrial locomotion in the ornithocheirids. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society, 2008, 154 , 27–69.     

Model of forced expiratory flows and airway geometry in infants.

Early measurements of autopsied lungs from infants, children, and adults suggested that the ratio of peripheral to central airway resistance was higher in infants than older children and adults. Recent measurements of forced expiration suggest that infants have high flows relative to lung volume. We employed a computational model of forced expiratory flow along with physiological and anatomic data to evaluate whether the infant lung is a uniformly scaled-down version of the adult lung. First, we uniformly scaled an existing computational model of adult forced expiration to estimate forced expiratory flows (FEF) and density dependence for an 18-mo-old infant. The values obtained for FEF and density dependence were significantly lower than those reported for healthy 18-mo-old infants. Next, we modified the model for the infant lung to reproduce standard indexes of expiratory flow [forced expiratory volume in 0.5 s (FEV(0.5)), FEFs after exhalation of 50 and 75% forced vital capacity, FEF between 25 and 75% expired volume] for this age group. The airway sizes obtained for the infant lung model that produced accurate physiological measurements were similar to anatomic data available for this age and larger than those in the scaled model. Our findings indicate that the airways in the infant lung model differ from those in the scaled model, i.e., middle and peripheral airway sizes are larger than result from uniform downscaling of the adult lung model. We show that the infant lung model can be made to reproduce individual flow-volume curves by adjusting lumen area generation by generation.