Characteristic Elongated Shape Research Articles

Definition and Quantification of Three-Dimensional Imaging Targets to Phenotype Pre-Eclampsia Subtypes: An Exploratory Study

Pre-eclampsia is a severe placenta-related complication of pregnancy with limited early diagnostic and therapeutic options. Aetiological knowledge is controversial, and there is no universal consensus on what constitutes the early and late phenotypes of pre-eclampsia. Phenotyping of native placental three-dimensional (3D) morphology offers a novel approach to improve our understanding of the structural placental abnormalities in pre-eclampsia. Healthy and pre-eclamptic placental tissues were imaged with multiphoton microscopy (MPM). Imaging based on inherent signal (collagen, and cytoplasm) and fluorescent staining (nuclei, and blood vessels) enabled the visualization of placental villous tissue with subcellular resolution. Images were analysed with a combination of open source (FIJI, VMTK, Stardist, MATLAB, DBSCAN), and commercially (MATLAB) available software. Trophoblast organization, 3D-villous tree structure, syncytial knots, fibrosis, and 3D-vascular networks were identified as quantifiable imaging targets. Preliminary data indicate increased syncytial knot density with characteristic elongated shape, higher occurrence of paddle-like villous sprouts, abnormal villous volume-to-surface ratio, and decreased vascular density in pre-eclampsia compared to control placentas. The preliminary data presented indicate the potential of quantifying 3D microscopic images for identifying different morphological features and phenotyping pre-eclampsia in placental villous tissue.

Relationship between nanotopographical alignment and stem cell fate with live imaging and shape analysis.

The topography of a biomaterial regulates cellular interactions and determine stem cell fate. A complete understanding of how topographical properties affect cell behavior will allow the rational design of material surfaces that elicit specified biological functions once placed in the body. To this end, we fabricate substrates with aligned or randomly organized fibrous nanostructured topographies. Culturing adipose-derived stem cells (ASCs), we explore the dynamic relationship between the alignment of topography, cell shape and cell differentiation to osteogenic and myogenic lineages. We show aligned topographies differentiate cells towards a satellite cell muscle progenitor state - a distinct cell myogenic lineage responsible for postnatal growth and repair of muscle. We analyze cell shape between the different topographies, using fluorescent time-lapse imaging over 21 days. In contrast to previous work, this allows the direct measurement of cell shape at a given time rather than defining the morphology of the underlying topography and neglecting cell shape. We report quantitative metrics of the time-based morphological behaviors of cell shape in response to differing topographies. This analysis offers insights into the relationship between topography, cell shape and cell differentiation. Cells differentiating towards a myogenic fate on aligned topographies adopt a characteristic elongated shape as well as the alignment of cells.

Lactoyl‐poloxamine/collagen matrix for cell‐containing tissue engineering modules

Collagen-containing crosslinked, remodelable poloxamine derivatives were produced by introducing very short oligo(lactic acid) segments through the reaction of poloxamine with L-lactide and the later addition of unsaturated bonds by the reaction of modified poloxamine with methacryloyl chloride. Degradation studies on discs indicated a faster weight loss in comparison to the stability of lactoyl-free samples. Cell-containing modules (both HepG2 cells and two different umbilical vein smooth muscle cell (UVSMC) cell-types) were produced. Live/Dead assay showed high survival levels for both HepG2 and UVSMC cell types after crosslinking. While nondegradable modules did not change shape over time, lactoyl-poloxamine matrices showed a gradual shrinkage and size decrease and an increase in the roughness of the surface. These findings were consistent with the expected degradability of the lactoyl derivative. A UVSMC cell line (CRL-2481) embedded in a LA-poloxamine/collagen matrix showed the characteristic elongated shape at day 9. UVSMC primary cells behaved in a manner similar to that seen in collagen gels: these cells formed isolated clusters through the matrix that gradually lost viability. On tissue culture polystyrene the same cells aggregated and did not reach confluence. Modules with embedded CRL-2481 UVSMC led to a better initial adhesion of endothelial cells and a higher extent of surface coverage than seen with the UVSMC-free system. With embedded primary UVSMC, some EC attachment and formation of gap junctions was seen. The pattern was not well organized. With further improvement (and characterization), the lactoyl poloxamine derivative is potentially useful as a scaffold for modular tissue engineering, when tissue remodeling is an important consideration.

Cartilage‐derived morphogenetic proteins induce osteogenic gene expression in the C2C12 mesenchymal cell line

Cartilage-derived morphogenetic protein-1, -2, and -3 (CDMP-1, -2, and -3) are members of the bone morphogenetic protein (BMP) family and have been shown to exhibit a variety of biological activities. In the present study, effects of these CDMPs on the temporal and spatial expression of genes in the pluripotent mesenchymal cell line C2C12 were examined. Cells cultured in the presence of CDMPs lost the characteristic elongated shape of myoblasts. At the molecular level, CDMP treatment did not change the mRNA expression of MyoD, aggrecan, Six1, and tendin. Scleraxis mRNA level was reduced by CDMP treatment. CDMP-1 and -3, but not CDMP-2, stimulated expression of osteogenic markers, such as alkaline phosphatase (AP), osteocalcin (OC), BSP, and type I collagen, in a dose- and time-dependent manner. With few exceptions, the three CDMPs changed, with different potencies, the expression profile of different members of the BMP family in a similar temporal pattern. Except at the late phase of treatment, CDMP treatment did not change the expression of ActR-IA, BMPR-IA, BMPR-IB, BMPR-II, and ALK-7 mRNAs. Based on the current data, the CDMPs appear to be able to stimulate the C2C12 cells to differentiate into the osteoblast pathway.

Stress measurements in high-shear granulators using calibrated “test” particles: application to scale-up

Granulation is a process by which fine powders are agglomerated into larger particles using a liquid binder. In high-shear granulation the powder–binder mix experiences intense agitation inside a mixing vessel as binder is dispersed and granules form and strengthen under the influence of shear and compacting forces in the device. It is an implicit assumption that in a “high shear” mixer, large forces are transmitted to the powder and this results in a short and efficient granulation process. Owing to these desirable characteristics, high-shear granulation was adopted by several industries including pharmaceuticals and detergents where the process is used almost exclusively. In the work reported here, we attempt to measure shear forces in a moving powder inside a mixer-granulator. The method is based on previous numerical simulations [Powder Technology 110 (2000) 59] and experiments [Journal of Fluid Mechanism 347 (1997) 347] where we showed that at equilibrium between stresses in the mixer and the yield strength of the particles, granules attain a characteristic elongated shape. The measuring method adopted is indirect in the sense that pellets with well-defined mechanical properties were used to interrogate forces inside the granulating vessel at the point where they attain their characteristic elongated shape. We subsequently used the condition of equal shear forces in the device as a scale-up criterion so as to preserve the magnitude of stresses at both scales and thereby to expose forming granules to similar forces in both the small- and large-scale machines. We found that shear forces in a “High-Shear” mixer-granulator with a vertical axis (Fielder) are actually not always high. The mixer has the potential to produce high shear forces but these forces are transmitted to the powder mass only if the powder is sufficiently cohesive or becomes cohesive due to binder addition. Shear forces in the granulator are strongly wet-mass-dependent and they increase rapidly as soon as a “granulation limit” is achieved, i.e., at the point where granules start to form in the shearing powder mass. We found that granulators with geometrically similar bowls can be scaled to generate comparable shear forces by decreasing the impeller rotational speed of the large machine by the factor ( D/ d) n , where D and d are the impeller diameters of the large and small machine, respectively, and n is a scaling index that depends on impeller geometry but not on wet mass properties. For the equipment studied in this work, the coefficient n was obtained as 0.80< n<0.85. We also propose an improved granulation process in which dry powders are pre-wetted before introduction into the main granulating device. This scheme has the potential to produce larger shear forces during wetting and binder introduction and thereby improve homogeneity and consequently final granule properties.

Experimental investigation of the transformation zone of tetragonal zirconia polycrystalline ceramics

Stress-induced martensitic transformation plastic zones of ceria-stabilized tetragonal zirconia polycrystalline ceramics, under some typical loading conditions, were studied by Moire interferometry. The full-field fringe patterns, including u-fields and v-fields, were acquired. According to these patterns, the transformation zone shape and transformation plasticity distributions of the specimens were obtained. The experimental results show that the stress-induced transformation at room temperature is concentrated in some narrow bands and the transformation plasticity is not uniform within the transformation zone. Experiments also reveal that the transformation zone with a characteristic elongated shape ahead of the notch, in a single-edge notch bending specimen, is obviously different from that resulting from some constitutive relation of transformation. This work provides a significant experimental foundation for establishing the theoretical models of transformation toughening.

An incompressible jet in a weak crossflow

A description of the incipient bending of a round incompressible jet issuing into a weak crossflow is presented. Axial vorticity is shown to appear from the early stages of the jet evolution owing to the distortion and reorientation of the azimuthal vorticity, and it eventually dominates the flow around the jet and determines the shape of its cross-section.The crossflow is considered weak enough for the distortion of the jet to occur downstream of its development region, where diffusion already influences the whole cross-section and the jet can be modelled as a point source of momentum. Axial pressure gradient and axial diffusion are negligible under these conditions, since the jet is a slender structure.Sufficiently near the origin, a finite-length entraining wake is identified on the lee side of the jet, which gradually merges with the main core. At the same time, the cross-section begins to acquire a characteristic elongated shape, with the jet concentrating in a thin layer. Farther downstream the axial vorticity of the jet rearranges into a couple of large locally two-dimensional contrarotating vortices standing against the wind under the action of their own induced velocity, and a smaller vortex sheet coinciding with the distorted jet.

Sub-domain structure of lipid-bound annexin-V resolved by electron image analysis

Two-dimensional crystals of annexin-V bound to lipid layers containing dioleoyl-phosphatidylserine have been obtained in the presence of Ca 2+. The crystals diffract to 20 Å resolution and have the symmetry of the plane group p3 (unit cell dimensions: a = b = 94 A ̊ , γ = 120 ° ). Electron image analysis revealed that the crystals are composed of trimers of annexin-V forming triskelion-like motifs. Each annexin-V molecule has a characteristic elongated shape, about 65 Å by 20 Å, when observed perpendicularly to the crystal plane. It is composed of two staggered domains of similar size, about 40 Å by 20 Å. Both domains are made of two sub-domains. The present data suggest that the four resolved sub-domains represent the folding units corresponding to the four 70 amino acid repeating segments characteristic of all annexins.

Transformation Zone Shape, Size, and Crack‐Growth‐Resistance [R‐Curve] Behavior of Ceria‐Partially‐Stabilized Zirconia Polycrystals

Transformation zone shape, size, and crack‐growth‐resistance (R‐curve) behavior were studied in precracked and annealed single‐edge notch bend specimens of commercial‐grade ceriapartially‐stabilized zirconia polycrystals as a function of applied load. Well‐defined transformation zones with a characteristic elongated shape in the plane of the crack were observed. It is shown that the observed zone shape is significantly different from the shape predicted by a combined shear/dilatation yield criterion and the stress field of the crack prior to the transformation. The length of the transformation zone directly ahead of the crack tip is in better agreement with the prediction of the Dugdale plastic strip zone model. The fracture toughness increment showed the characteristic square root dependence on the transformation zone width, but the magnitude of the toughness increment was not consistent with the predictions of the theoretical models of transformation toughening.