The role of Mg++-ATPase (actomyosin-like protein) in maintaining the biconcave shape of erythrocytes.

Chemically different substances known to change the Mg++-ATPase activity in the red cell membrane, likewise alter the red cell shape. Normal human red cells retain their biconcave shape only when the activity of this enzyme remains unchanged. The present work deals with the possibility that Mg++-ATPase may cause certain tension in the membrane responsible for the biconcave shape of the erythrocyte.     

A methodology for estimating the shape of biconcave red blood cells using multicolor scattering images

In this paper, a novel methodology for estimating the shape of human biconcave red blood cells (RBCs), using color scattering images, is presented. The information retrieval process includes, image normalization, features extraction using two-dimensional discrete transforms, such as angular radial transform (ART), Zernike moments and Gabor filters bank and features dimension reduction using both independent component analysis (ICA) and principal component analysis (PCA). A radial basis neural network (RBF-NN) estimates the RBC geometrical properties. The proposed method is evaluated in both regression and identification tasks by processing images of a simulated device used to acquire scattering phenomena of moving RBCs. The simulated device consists of a tricolor light source (light emitting diode – LED) and moving RBCs in a thin glass. The evaluation database includes 23,625 scattering images, obtained by means of the boundary element method. The regression and identification accuracy of the actual RBC shape is estimated using three feature sets in the presence of additive white Gaussian noise from 60 to 10 dB SNR and systematic distortion, giving a mean error rate less than 1% of the actual RBC shape, and more than 99% mean identification rate.     

2D-finite element analyses and histomorphology of lag screws with and without a biconcave washer.

For osteosynthesis and for bone transplant fixation in particular, a lag screw with a biconcave washer, the so called "Anchor Screw" (AS) has been introduced in maxillo-facial surgery. Using 2D-finite element analysis (FEA), the v.Mises and the circumferential stresses induced in underlying bone by this AS are analysed and compared to those under a conventional lag screw. The stress distributions below the biconcave washer of the AS were correlated with histomorphological bone reactions after AS osteosynthesis in two tumor patients, retrieved 12 weeks and 19 months after tumor surgery, respectively. Depending on the thickness of cortical bone, the v.Mises stress concentrations below the biconcave washer were lower than under the head of the conventional lag screw (CLS), but with a higher stress maximum concentrated around the rim of the washer. The circumferential stresses were only half as high around the AS, and thus the deformation of bone was reduced. As predicted by FEA, histology showed microcrack formation, but then after minimal resorption, remodelling of bone below the biconcave washer. Stable osteosynthesis could be demonstrated by bony union already after 12 weeks, and, while bone remodelling continued in the healed osteotomy, it had decreased around the screws after 19 months. It can be concluded from the biomechanical principles and the histomorphological findings that the AS appears superior to the CLS.     

Thermohemolysis of erythrocytes

Thermohemolysis kinetic curves of erythrocytes in the temperature interval of 37-75 degrees C in different media and in the course of blood storage were studied. Mechanism of thermohemolysis is proposed. The rate constants of hemolysis stages are introduced. It is shown that these parameters are sensitive to the changes of structural state of erythrocytes.     

Microencapsulation of erythrocytes

Human erythrocytes have been encapsulated in a polyacrylate membrane by a simple precipitation process. The encapsulated cells appeared to remain functional after encapsulation: the consumption of glucose and the ability to reversibly bind oxygen was unimpaired. Furthermore, storage at 4°C for almost 6 months had no effect on the P₅₀ and n₅₀ values. This is the first time to our knowledge that live mammalian cells have been encapsulated in a polymer other than alginate.     

Sequence representation

This article deals with the definition of a method for analyzing sequences of symbols, especially biological sequences. We are mostly interested in finding representations of sequences, that could help to explicit relationship between their structure and their activity. Starting with automatically built rules, governing occurrences of symbols within sequences, we define ways of using these rules to determine different subsequences that we assume to be contexts. Labelled contexts provide a possible representation of sequences. Finally an example of detected contexts in proteins is given.     

Is there any connection between heat inactivation of spectrin-dependent ATPase and loss of smooth biconcave shape of red cells?

Spectrin-dependent ATPase activity was measured in membranes from native human erythrocytes and erythrocytes heated for 20 min at different temperatures. This activity was found to decline when the erythrocytes were heated at 48 degrees C and higher. The break in ATPase activity corresponds to morphological changes in erythrocytes found by Crome and Mollison [Brit. J. Haematol. (1964) 10, 137]. The role of spectrin-dependent ATPase in erythrocyte shape maintenance is discussed.     

Flicker spectroscopy of erythrocytes

Frequency analysis of thermally excited surface undulations of erythrocytes leading to the flicker phenomenon is applied to determine biochemically and physically induced modulations of the membrane curvature elasticity. Flicker spectra of individual cells fixed to the window of a flow chamber by polylysine are taken by phase contrast microscopy, enabling investigations of the reversibility of the structural modifications. The spectra may be approximated by Lorentzian lines in most cases. By measuring the amplitude (at zero frequency) and the line width, effects of the structural changes on the curvature elastic constant, K _c , and the wavelength distribution of the undulations may be studied separately. Effect of physically induced modifications: The temperature dependence of the flicker spectra are taken from 10°C to 37°C. Above 20°C, K _c decreases with increasing temperature whereas the reverse holds below this limit. The latter anomalous behaviour is explained in terms of a conformational change associated with protein and lipid lateral phase separation. The bending stiffness increases when the cells swell osmotically, owing to surface tension effects. The dependence of the flicker spectra on the viscosity of the suspension medium agrees with the theoretical prediction. Biochemically and drug induced modifications: 5 vol of ethanol leads to a pronounced and reversible suppression of the long wavelength undulations without altering the discoid cell shape and without affecting the bending stiffness appreciably. Adsorption of dextran to the glycocalix increases K _c by a factor of 1.6 at saturation. The bending stiffness is increased by a factor of 1.3 after cross-linking the proteins with the SH-oxidizing agent diamid. Injection of Ca⁺⁺ into the cell via ionophores evokes (within 10 min) the formation of fine — probably spectrin free — spicules. This leads to an increase in K _c by a factor of 1.3 which is explained in terms of a lateral condensation of the spectrin/actin network. The spicule formation and K _c change is completely reversible (within 2 min) after perfusion with Ca⁺⁺-free buffer. Cholesterol depletion leads first to a continuous increase in K _c without change of the cell shape whereas a sudden discocyte- to echinocyte transformation sets in below a critical steroid content. The latter transition is also observed in cell suspensions and is reminiscent of a phase transition. The anti-tumor drug actinomycin D evokes an increase in the bending stiffness K _c by a factor of two, suggesting that its effect is at least partially due to a modulation of the membrane structure. The -receptor agonist leads to a remarkable increase in K _c (by about 25%) at 10^-4 M but the effect is not reversed by the -antagonist prazosin, suggesting that the agonist exerts a non-specific effect.A new technique, dynamic reflection interference contrast microscopy, is introduced by which absolute values of the amplitudes of the surface undulations and therefore K _c can be determined. The value obtained: K _c =5·10^-13 erg is about a factor of two larger then the bending stiffness of pure lipid bilayers. We suggest that the surface undulations may also be determined by lateral fluctuations of the quasi-two-dimensional spectrin/actin network.     

Ultrastructure of malaria-infected erythrocytes

Knobs, caveolae, caveola-vesicle complexes, cytoplasmic clefts, and electron-dense material are five major ultrastructural changes found in the membrane skeleton and cytoplasm of erythrocytes infected with species of primate malaria. Knobs are electron-dense, conical evaginations of the erythrocyte surface, which are believed to mediate cytoadherence and sequestration of Plasmodium falciparum-infected erythrocytes. Caveolae and caveola-vesicle complexes are flask-shaped invaginations of the membrane skeleton, which may be involved in the uptake or export of host- or parasite-derived substances. Cytoplasmic clefts are flattened or circular membranous structures found in the erythrocyte cytoplasm between the intracellular parasite and the host cell surface. The clefts are variable in length and bounded by two or more membranes. Fine, granular electron-dense material is often found on the cytoplasmic face of clefts or in amorphous packets in the erythrocyte cytoplasm. Immunocytochemistry has demonstrated that all of these ultrastructural changes are associated with the trafficking and interaction of specific malarial antigens with the host erythrocyte.