Analysis of the formation of parallel arrays by BHK cells in vitro

The contact behavior of BHK cells in tissue culture was studied in order to elucidate the mechanism for their formation of parallel arrays. It was found that when two arrays of cells meet at angles of less than 55 °, they merge and form a single array. At angles of contact greater than 55 °, these arrays crisscross. This same behavior was noted when two individual cells contacted each other. At angles up to 55 ° the contacting cell alters its direction of movement so that it comes to lie parallel to the contacted cell. At angles greater than 55 °, in contrast, crisscrossing occurs. Detailed analysis at high magnification showed that this behavior is dependent upon localized contact inhibition of cell movement.     

Membrane glycoprotein and surface free energy changes in hypoxic fibroblast cells

Hypoxia affects the biochemistry of mammalian cells and thus alters their sensitivity to subsequent chemo- and radiotherapy. When V79 Chinese hamster lung fibroblasts were grown under conditions of extreme hypoxia (less than 10 ppm O2) there was a significant shift in the membrane glycoprotein composition. Scanning electron microscopy revealed altered cell surface morphology including loss of pseudopodial projections. Experiments to determine changes in interfacial free energy of these cells using equilibrium two phase systems of poly(ethylene glycol) (PEG) and dextran were carried out. Test fluid droplets of the denser dextran-rich phase were formed on layers of cells in the PEG-rich phase as the bathing medium, and the contact angles the droplets made with the cell layers were measured from photomicrographs. The contact angles on cells in the plateau phase increased significantly with time of exposure to hypoxia, from 25 degrees (zero time) to 35 degrees (6 h) to 60 degrees (9 h). Contact angles on cells in the exponential phase increased from 80 degrees (zero time) to 150 degrees after 20 h of hypoxia. It appears that the altered contact angles reflect changes in cell surface hydrophobicity that may, in part, reflect alterations in the membrane glycoprotein composition.     

Fine structure of wall bodies in large cells of an Anabaena species.

A new type of inclusion in blue-green algae termed a wall body is described. These are elongate bodies of medium electron density, from 6 to 70 nm in thickness and at least 0.2 micrometer in depth found in large cells of Anabaena sp. B387. The bodies are up to 12 micrometer long, extending the complete length of the cell in some cases. Some of the bodies are bifurcated and the ends appeared to come in contact with the plasma membrane. The wall bodies lie at various angles in the cells. Up to six have been observed in a single thin section of a cell. The large cells are found in greater numbers in cultures grown in a medium solidified with agar. Comparison of the bodies to the mucopolymer layer (layer 2) of the cell wall is made.     

Surface Hydrophobicity of Culture and Water Biofilm of <Emphasis Type="Italic">Penicillium</Emphasis> spp.

Fungal surface hydrophobicity is involved in several functions in fungal growth and development. Water contact angles measurement has been used as a direct and simple approach for its characterisation in solid cultures. Microsphere adhesion assay is said to be the best method to assess cell hydrophobicity of filamentous fungi. This study aimed to apply these two methods to study hydrophobicity of Penicillium expansum and Penicillium brevicompactum grown as mycelial mats in solid culture, liquid culture and water biofilms. As result, both species in solid cultures were classified as hydrophobic with contact angles ≥90o, but in liquid cultures and water biofilms showed different levels of hydrophobicity when microsphere adhesion assay was applied. In addition, was found that biofilms have specific hydrophobic hyphae which may be involved in fungal ecological functions.     

Non-straight cell edges are important to invasion and engulfment as demonstrated by cell mechanics model

Computational models of cell–cell mechanical interactions typically simulate sorting and certain other motions well, but as demands on these models continue to grow, discrepancies between the cell shapes, contact angles and behaviours they predict and those that occur in real cells have come under increased scrutiny. To investigate whether these discrepancies are a direct result of the straight cell–cell edges generally assumed in these models, we developed a finite element model that approximates cell boundaries using polylines with an arbitrary number of segments. We then compared the predictions of otherwise identical polyline and monoline (straight-edge) models in a variety of scenarios, including annealing, single- and multi-cell engulfment, sorting, and two forms of mixing—invasion and checkerboard pattern formation. Keeping cell–cell edges straight influences cell motion, cell shape, contact angle, and boundary length, especially in cases where one cell type is pulled between or around cells of a different type, as in engulfment or invasion. These differences arise because monoline cells have restricted deformation modes. Polyline cells do not face these restrictions, and with as few as three segments per edge yielded realistic edge shapes and contact angle errors one-tenth of those produced by monoline models, making them considerably more suitable for situations where angles and shapes matter, such as validation of cellular force–inference techniques. The findings suggest that non-straight cell edges are important both in modelling and in nature.     

The role of bacterial cell wall hydrophobicity in adhesion.

In this study, the adhesion of bacteria differing in surface hydrophobicity was investigated. Cell wall hydrophobicity was measured as the contact angle of water on a bacterial layer collected on a microfilter. The contact angles ranged from 15 to 70 degrees. This method was compared with procedures based upon adhesion to hexadecane and with the partition of cells in a polyethylene glycol-dextran two-phase system. The results obtained with these three methods agreed reasonably well. The adhesion of 16 bacterial strains was measured on sulfated polystyrene as the solid phase. These experiments showed that hydrophobic cells adhered to a greater extent than hydrophilic cells. The extent of adhesion correlated well with the measured contact angles (linear regression coefficient, 0.8).     

Effects of cell surface damage on surface properties and adhesion of Pseudomonas aeruginosa

Bacterial cell surfaces play a crucial role in their adhesion to surfaces. In the present study, physico-chemical cell surface properties of Pseudomonas aeruginosa, isolated from a case of contact lens associated keratitis, are determined for mid-exponential and early stationary phase cells and for cells after exposure to a lens care solution or after mechanical damage by sonication. Exposure to a lens care solution and mechanical cell surface damage reduced the cell surface hydrophobicity and water contact angles decreased from 129 degrees to 96 degrees and 83 degrees, respectively. Zeta potentials in saline (-9 mV) were hardly affected after mechanical damage, but tri-modal zeta potential distributions, with subpopulation zeta potentials at -11, -28 and -41 mV, were observed after exposure of bacteria to a lens care solution. X-ray photoelectron spectroscopy indicated changes in the amounts of oxygen-, nitrogen- and phosphorus-rich cell surface components. Mid-exponential phase cells had more nitrogen-rich cell surface components than early stationary phase cells, but water contact angles and zeta potentials were not very different. In addition, mid-exponential phase cells adhered better than early stationary phase cells to hydrophobic and hydrophilic substrata in a parallel plate flow chamber. The capacity of P. aeruginosa to adhere was decreased after inflicting cell surface damage. Exposure to a lens care solution yielded a larger reduction in adhesion capacity than sonication, likely because sonication left most of the cells in a viable state, in contrast to exposure to a lens care solution. It is argued that for clinically relevant experiments, it may be preferable to work with surface damaged cells rather than with gently harvested organisms.     

Orientation of the cleavage spindles in pulmonate mollusks. I. The role of the form of the blastomeres in 2d cleavage spindle orientation

In the normal two-celled embryos of various pulmonate molluscs, the orientation of spindles characteristic of metaanaphase is being frequently established gradually, in the process of transition from pro- to metaphase accompained by the growth of spindle and asters. The typical growth of contact zone between the blastomeres of the common pond snail embryos was inhibited to a different extent under their cultivation after the 1 cleavage division in the calcium-free media or after trypsinization. At the same time the orientation of meta-anaphase spindles was markedly affected (as judged by an angle alpha between the spindle axis and the plane of contact zone in the equatorial projection). When analyzing the model distributions of the angles between the two spindle axes (in the same projection), it was shown that the empirical distributions of these angles corresponded to the principle of stochastic combination of two alpha. A conclusion is drawn that the orientation of one spindle does not depend on that of another but the position of each of them depends on the size of the contact zone and, hence, on the general form of the adjacent blastomere region. Some other processes determining the spindle orientation are discussed.     

Effect of knee joint flexion and femur rotation on retropatellar contact of the human knee joint]

The aim of the present study was to evaluate retropatellar contact characteristics at different angles of flexion of the knee joint. To this end, 6 cadaveric legs were examined using pressure sensitive film (Fuji Prescale type "super low") at angles of flexion of 45 degrees, 60 degrees, 90 degrees and 120 degrees both in neutral rotation and 10 degrees internal and external rotation of the femur in the same knee joints. A force of 140 N was applied to both the vastus medialis and lateralis, and a comparison made with a medially and a laterally dominating muscle force. The contact areas decreased with increasing angles of flexion. The medially dominating muscle traction increased the contact area. Comparison between internal and external rotation revealed a decrease in contact area on internal rotation. The pressure measurements were comparable in all loading situations. Comparison between neutral and medial traction revealed significant differences in contact area, pressure and force. The influence of femoral rotation showed no significant difference. A comparison of the different angles of flexion revealed only few significant differences. To prevent the development of retropatellar arthrosis, maximum contact areas are necessary. The study has shown an advantage for medially dominating muscle traction, and external rotation of the femur.     

A two-phase growth strategy in cultured neuronal networks as reflected by the distribution of neurite branching angles

Neurite outgrowth and branching patterns are instrumental in dictating the wiring diagram of developing neuronal networks. We study the self-organization of single cultured neurons into complex networks focusing on factors governing the branching of a neurite into its daughter branches. Neurite branching angles of insect ganglion neurons in vitro were comparatively measured in two neuronal categories: neurons in dense cultures that bifurcated under the presence of extrinsic (cellular environment) cues versus neurons in practical isolation that developed their neurites following predominantly intrinsic cues. Our experimental results were complemented by theoretical modeling and computer simulations. A preferred regime of branching angles was found in isolated neurons. A model based on biophysical constraints predicted a preferred bifurcation angle that was consistent with this range shown by our real neurons. In order to examine the origin of the preferred regime of angles we constructed simulations of neurite outgrowth in a developing network and compared the simulated developing neurons with our experimental results. We tested cost functions for neuronal growth that would be optimized at a specific regime of angles. Our results suggest two phases in the process of neuronal development. In the first, reflected by our isolated neurons, neurons are tuned to make first contact with a target cell as soon as possible, to minimize the time of growth. After contact is made, that is, after neuronal interconnections are formed, a second branching strategy is adopted, favoring higher efficiency in neurite length and volume. The two-phase development theory is discussed in relation to previous results.     

The effects of superficial wax on leaf wettability

Experiments are described which provide more information on the role played by superficial waxes in the natural water-repellency of leaf surfaces. Contact angles of water were measured on a variety of leaf surfaces, before and after removal of wax, and on smooth films of the isolated superficial waxes. The differences in wettability of leaf surfaces are not wholly accounted for by differences which occur in the chemical and hydrophobic properties of their superficial waxes. Waxes isolated from leaves exhibiting contact angles less than 90° are usually more hydrophobic than the leaf surface itself. On most leaves exhibiting angles greater than 90° wax is the dominant factor governing water-repellency, the isolated wax normally making at least a 60 % contribution to the contact angle measured on the leaf surface. Additional factors, such as roughness, responsible for the occurrence of contact angles greater than 110° on certain leaf surfaces, reside in the wax layer. The hydrophobic properties of some leaves are unaffected by chloroform washing, revealing that superficial waxes play little part in their wettability.     

A novel method for identifying hydrophobicity on fungal surfaces

Fungal surface hydrophobicity has many ecological functions and water contact angles measurement is a direct and simple approach for its characterization. The objective of this study was to evaluate if in-vitro growth conditions coupled with versatile image analysis allows for more accurate fungal contact angle measurements. Fungal cultures were grown on agar slide media and contact angles were measured utilizing a modified microscope and digital camera setup. Advanced imaging software was adopted for contact angle determination. Contact angles were observed in hydrophobic, hydrophilic and a newly created chronoamphiphilic class containing fungi taxa with changing surface hydrophobicity. Previous methods are unable to detect slight changes in hydrophobicity, which provide vital information of hydrophobicity expression patterns. Our method allows for easy and efficient characterization of hydrophobicity, minimizing disturbance to cultures and quantifying subtle variation in hydrophobicity.     

A two‐phase growth strategy in cultured neuronal networks as reflected by the distribution of neurite branching angles

Neurite outgrowth and branching patterns are instrumental in dictating the wiring diagram of developing neuronal networks. We study the self‐organization of single cultured neurons into complex networks focusing on factors governing the branching of a neurite into its daughter branches. Neurite branching angles of insect ganglion neurons in vitro were comparatively measured in two neuronal categories: neurons in dense cultures that bifurcated under the presence of extrinsic (cellular environment) cues versus neurons in practical isolation that developed their neurites following predominantly intrinsic cues. Our experimental results were complemented by theoretical modeling and computer simulations. A preferred regime of branching angles was found in isolated neurons. A model based on biophysical constraints predicted a preferred bifurcation angle that was consistent with this range shown by our real neurons. In order to examine the origin of the preferred regime of angles we constructed simulations of neurite outgrowth in a developing network and compared the simulated developing neurons with our experimental results. We tested cost functions for neuronal growth that would be optimized at a specific regime of angles. Our results suggest two phases in the process of neuronal development. In the first, reflected by our isolated neurons, neurons are tuned to make first contact with a target cell as soon as possible, to minimize the time of growth. After contact is made, that is, after neuronal interconnections are formed, a second branching strategy is adopted, favoring higher efficiency in neurite length and volume. The two‐phase development theory is discussed in relation to previous results. © 2004 Wiley Periodicals, Inc. J Neurobiol, 2005     

Influence of epiphytic micro-organisms on leaf wettability: wetting of the upper leaf surface of Juglans regia and of model surfaces in relation to colonization by micro-organisms

Wetting of the upper leaf surface of Juglans regia L. and of model surfaces colonized by epiphytic micro-organisms was investigated by measuring contact angles of aqueous solutions buffered at different pH values. During June to October 1995, contact angles of aqueous solutions on the leaf surface of J. regia decreased by angles ranging from 12° (low pH values) to 25° at high pH values. At the end of this vegetation period, wetting was strongly dependent on pH showing significantly lower contact angles with alkaline solutions (pH 9·0) than with acidic solutions (pH 3·0). Contact angle titration measured angles on the leaf surface as a function of the pH of buffered aqueous solutions, covering a pH range from 3·0 to 11·0. Titration curves revealed inflection points around 7·5, indicating the existence of ionizable carboxylic groups at the interface of the phylloplane. Altered leaf-surface wetting properties observed on the intact leaf surface could be simulated in model experiments by measuring contact angles on artificial surfaces colonized by Pseudomonas fluorescens and by epiphytic micro-organisms isolated from the phylloplane of J. regia . Strong evidence is provided that interfacial carboxylic groups derive from epiphytic micro-organisms present on the phylloplane. Results suggest that the age-dependent increase in, and pH dependence of, wetting as leaves mature are related to the presence of epiphytic micro-organisms on the phylloplane. Ecological consequences of increased leaf-surface wetting, concerning the structure of the leaf surface as a microhabitat for epiphytic micro-organisms, are discussed.     

A kinematic model of the human hand to evaluate its prehensile capabilities.

A kinematic model has been developed for simulation and prediction of the prehensile capabilities of the human hand. The kinematic skeleton of the hand is characterized by ideal joints and simple segments. Finger-joint angulation is characterized by yaw (abduction-adduction), pitch (flexion-extension) and roll (axial rotation) angles. The model is based on an algorithm that determines contact between two ellipsoids, which are used to approximate the geometry of the cutaneous surface of the hand segments. The model predicts the hand posture (joint angles) for power grasp of ellipsoidal objects by 'wrapping' the fingers around the object. Algorithms for two grip types are included: (1) a transverse volar grasp, which has the thumb abducted for added power; and (2) a diagonal volar grasp, which has the thumb adducted for an element of precision. Coefficients for estimating anthropometric parameters from hand length and breadth are incorporated in the model. Graphics procedures are included for visual display of the model. In an effort to validate the predictive capabilities of the model, joint angles were measured on six subjects grasping circular cylinders of various diameters and these measured joint angles were compared with angles predicted by the model. Sensitivity of the model to the various input parameters was also determined. On an average, the model predicted joint flexion angles that were 5.3% or 2.8 degrees +/- 12.2 degrees larger than the measured angles. Good agreement was found for the MCP and PIP joints, but results for DIP were more variable because of its dependence on the predictions for the proximal joints.     

Determination of contact angle by molecular simulation using number and atomic density contours

The contact angles of Lennard-Jones fluid droplets on a structureless solid surface, simulated using Monte Carlo simulation, are calculated by fitting isochoric surfaces and making a number of assumptions about the droplet. The results show that there are significant uncertainties in the calculated contact angles due to the choice of these assumptions, such as the grid size used in tracking the isochoric density profile, the omission of isochoric data points near the surface and the function used to fit the isochoric profile. In this study, we propose a new method of calculating density contours based on atomic density instead of number density. This method results in a much smaller variation in contact angle when applying different assumptions than using number density for isochoric contours. The most consistent results, across a range of assumptions about the droplet and the contact angle, come from averaging the contact angle from several isochoric density profiles. In addition, this gives a measurement of the variation due to the choice of isochoric density.     

The effect of vastus medialis forces on patello-femoral contact: a model-based study

A mathematical model of the patello-femoral joint was introduced to investigate the impact of the vastus medialis (longus, obliquus) forces on the lateral contact force levels. In the model, the quadriceps were represented as five separate forces: vastus lateralis, vastus intermedius, rectus femoris, vastus medialis longus (VML), and obliquus (VMO). By varying the relative force generation ratios of the quadriceps heads, the patello-femoral contact forces were estimated. We sought to analytically determine the range of forces in the VMO and VML that cause a reduction or an increase of lateral contact forces, often the cause of patello-femoral pain. Our results indicated that increased contact forces are more dependent on combinations of muscle forces than solely VMO weakness. Moreover, our simulation data showed that the contact force levels are also highly dependent on the knee flexion angle. These findings suggest that training targeted to reduce contact forces through certain joint angles could actually result in a significant increase of the contact forces through other joint angles.     

Mapping fluorophore distributions in three dimensions by quantitative multiple angle-total internal reflection fluorescence microscopy.

The decay of evanescent field intensity beyond a dielectric interface depends upon beam incident angle, enabling the 3-d distribution of fluorophores to be deduced from total internal reflection fluorescence microscopy (TIRFM) images obtained at multiple incident angles. Instrumentation was constructed for computer-automated multiple angle-TIRFM (MA-TIRFM) using a right angle F2 glass prism (n(r) 1.632) to create the dielectric interface. A laser beam (488 nm) was attenuated by an acoustooptic modulator and directed onto a specified spot on the prism surface. Beam incident angle was set using three microstepper motors controlling two rotatable mirrors and a rotatable optical flat. TIRFM images were acquired by a cooled CCD camera in approximately 0.5 degree steps for >15 incident angles starting from the critical angle. For cell studies, cells were grown directly on the glass prisms (without refractive index-matching fluid) and positioned in the optical path. Images of the samples were acquired at multiple angles, and corrected for angle-dependent evanescent field intensity using "reference" images acquired with a fluorophore solution replacing the sample. A theory was developed to compute fluorophore z-distribution by inverse Laplace transform of angle-resolved intensity functions. The theory included analysis of multiple layers of different refractive index for cell studies, and the anisotropic emission from fluorophores near a dielectric interface. Instrument performance was validated by mapping the thickness of a film of dihexyloxacarbocyanine in DMSO/water (n(r) 1.463) between the F2 glass prism and a plano-convex silica lens (458 mm radius, n(r) 1.463); the MA-TIRFM map accurately reproduced the lens spherical surface. MA-TIRFM was used to compare with nanometer z-resolution the geometry of cell-substrate contact for BCECF-labeled 3T3 fibroblasts versus MDCK epithelial cells. These studies establish MA-TIRFM for measurement of submicroscopic distances between fluorescent probes and cell membranes.     

Surface tension measurements on filamentous broths

Summary The surface tension of microorganisms is an important parameter in biotechnological processes. In this work contact angles and surface tensions were determined for filamentous micro-organisms Streptomyces levoris and Aspergillus niger. The contact angles were 39.7° and 43.1°, and the surface tensions 59.0 mJ/m² and 57.3 mJ/m², respectively. The contact angles for the filamentous broths were measured with the axisymmetric drop shape analysis - contact diameter (ADSA-CD) method which is particularly well suited to rough surfaces. The contact angle results obtained were significantly larger than those earlier reported for bacteria.Visiting scientist from Helsinki University of Technology, Espoo, Finland.     

Finite element prediction of contact pressures in cam-type femoroacetabular impingement with varied alpha angles

Three dimensional finite element models of cam-type FAI with alpha angles of 60°, 70°, 80°, and 90° were created to investigate the cartilage contact mechanics in daily activities. Intra-articular cartilage contact pressures during routine daily activities were assessed and cross-compared with a normal control hip. Alpha angles and hip range of motion were found to have a combined influence on the cartilage contact mechanics in hips with cam-type FAI, thereby resulting in abnormally high pressures and driving the cartilage damage. In particular, alpha angles of 80° or greater contribute to substantial pressure increase under certain types of daily activities.