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PubMed1999

Calculation of a Gap restoration in the membrane skeleton of the red blood cell: possible role for myosin II in local repair.

Human red blood cells contain all of the elements involved in the formation of nonmuscle actomyosin II complexes (V. M. Fowler. 1986. J. Cell. Biochem. 31:1-9; 1996. Curr. Opin. Cell Biol. 8:86-96). No clear function has yet been attributed to these complexes. Using a mathematical model for the structure of the red blood cell spectrin skeleton (M. J. Saxton. 1992. J. Theor. Biol. 155:517-536), we have explored a possible role for myosin II bipolar minifilaments in the restoration of the membrane skeleton, which may be locally damaged by major mechanical or chemical stress. We propose that the establishment of stable links between distant antiparallel actin protofilaments after a local myosin II activation may initiate the repair of the disrupted area. We show that it is possible to define conditions in which the calculated number of myosin II minifilaments bound to actin protofilaments is consistent with the estimated number of myosin II minifilaments present in the red blood cells. A clear restoration effect can be observed when more than 50% of the spectrin polymers of a defined area are disrupted. It corresponds to a significant increase in the spectrin density in the protein free region of the membrane. This may be involved in a more complex repair process of the red blood cell membrane, which includes the vesiculation of the bilayer and the compaction of the disassembled spectrin network.

Actins↗

PubMedDate not supplied

Spectrin properties and the elasticity of the red blood cell membrane skeleton.

Two models of spectrin elasticity are developed and compared to experimental measurements of the red blood cell (RBC) membrane shear modulus through the use of an elastic finite element model of the RBC membrane skeleton. The two molecular models of spectrin are: (i) An entropic spring model of spectrin as a flexible chain. This is a model proposed by several previous authors. (ii) An elastic model of a helical coiled-coil which expands by increasing helical pitch. In previous papers, we have computed the relationship between the stiffness of a single spectrin molecule (K) and the shear modulus of a network (mu), and have shown that this behavior is strongly dependent upon network topology. For realistic network models of the RBC membrane skeleton, we equate mu to micropipette measurements of RBCs and predict K for spectrin that is consistent with the coiled-coli molecular model. The value of spectrin stiffness derived from the entropic molecular model would need to be at least 30 times greater to match the experimental results. Thus, the conclusion of this study is that a helical coiled-coil model for spectrin is more realistic than a purely entropic model.

Cytoskeleton↗

PubMed2002

Monolithic silica columns with various skeleton sizes and through-pore sizes for capillary liquid chromatography.

Reduction of through-pore size and skeleton size of a monolithic silica column was attempted to provide high separation efficiency in a short time. Monolithic silica columns were prepared to have various sizes of skeletons (approximately 1-2 microm) and through-pores (approximately 2-8 microm) in a fused-silica capillary (50-200 microm I.D.). The columns were evaluated in HPLC after derivatization to C18 phase. It was possible to prepare monolithic silica structures in capillaries of up to 200 microm I.D. from a mixture of tetramethoxysilane and methyltrimethoxysilane. As expected, a monolithic silica column with smaller domain size showed higher column efficiency and higher pressure drop. High external porosity (> 80%) and large through-pores resulted in high permeability (K = 8 x 10(-14) -1.3 x 10(-12) m2) that was 2-30 times higher than that of a column packed with 5-mirom silica particles. The monolithic silica columns prepared in capillaries produced a plate height of about 8-12 microm with an 80% aqueous acetonitrile mobile phase at a linear velocity of 1 mm/s. Separation impedance, E, was found to be as low as 100 under optimum conditions, a value about an order of magnitude lower than reported for conventional columns packed with 5-microm particles. Although a column with smaller domain size generally resulted in higher separation impedance and the lower total performance, the monolithic silica columns showed performance beyond the limit of conventional particle-packed columns under pressure-driven conditions.

Chromatography, Liquid↗

PubMed1998

Mobilization of lead from the skeleton during the postnatal period is larger than during pregnancy.

A cohort of 15 immigrant females to Australia and 7 native Australian controls were monitored on a monthly basis with high-precision lead isotopic methods during gestation and for 6 months after pregnancy to determine the extent of lead mobilization from the maternal skeleton. Quarterly environmental samples of house dust, drinking water, urban air, gasoline, and a 6-day duplicate diet were also measured. The geometric mean blood lead concentration for the immigrant females on arrival in Australia was 3.0 microg/dl (range: 1.9 to 20 microg/dl), and for the Australian controls was 3.1 gm/dl (range: 1.9 to 4.3 microg/dl). During gestation and after pregnancy, blood lead concentrations varied, with mean individual changes of -14% to 83%. For the immigrant subjects, the percentage change in blood lead concentration was significantly greater during the postpregnancy period than during the 2nd and 3rd trimesters (p < 0.001). Skeletal contribution to blood lead, based on the isotopic composition for the immigrant subjects, increased in an approximately linear manner during pregnancy. The mean increases for each individual during pregnancy varied from 26% to 99%. Skeletal lead contribution to blood lead was significantly greater (p < 0.001) during the postpregnancy period than during the 2nd and 3rd trimesters. The contribution of skeletal lead to blood lead during the postpregnancy period remained essentially constant at the increased level of lead mobilization, although the duration of breastfeeding varied from 1 week to more than 6 months. The increased contribution of skeletal lead to blood lead during the postpregnancy period is attributed to increased mobilization of lead from maternal skeletal stores during lactation. The increased contribution of skeletal lead both during pregnancy and in the postpregnancy period is consistent with increased bone resorption, and may be associated with an inadequate calcium intake observed in quarterly 6-day duplicate diets. Mobilization of skeletal lead stores represents a potentially important source of perinatal lead intake and accumulation in the developing fetus. Only two subjects consumed dietary supplements for calcium, and their mobilization of lead from the skeleton to the blood was the lowest of all the subjects. These two subjects' use of calcium supplements may have reduced mobilization of skeletal mineral stores to supply the calcium needs of pregnancy and lactation. Calcium supplementation may be an important means of limiting fetal exposure to lead.

Bone and Bones↗

PubMed1999

Estimation of cumulative lead releases (lead flux) from the maternal skeleton during pregnancy and lactation.

Recent longitudinal studies with human subjects and nonhuman primates using high-precision stable lead isotopes show that lead is mobilized from the maternal skeleton during pregnancy and the postpartum period. We have now calculated the cumulative lead release (lead flux in micrograms) mobilized from the skeleton during these periods by means of analysis of monthly PbB samples from recent immigrants to Australia. Results included a statistically significant inverse relationship (P = .006) between the lead flux and the time of conception after the arrival of the subjects in Australia. By using an area-under-the-curve approach to determine the added lead inputs to blood during pregnancy and nursing versus a baseline value, the net lead release to blood varied from 0.9 to 10.1 microg/d, which is equivalent to 0.3 to 4.03 mg of lead. With group PbB concentrations usually less than 3 microg/dL, the observed releases imply a high skeletal turnover of greater than 10% and possibly greater than 30% in some subjects during pregnancy and the postpartum period. These elevated rates in some subjects may partly arise from low daily calcium intakes, being one half to two thirds of that of recommended daily requirements. The lead flux calculated from a cumulative approach was compared with other approaches: first-order kinetics, bone turnover, bone x-ray fluorescence measurements, and the International Commission for Radiological Protection lead pharmacokinetic model. Calculated lead releases and remaining bone lead concentrations would likely not be detectable by current x-ray fluorescence methods.

Adolescent↗

PubMed1989

Computer simulation of the hydrostatic skeleton. The physical equivalent, mathematics and application to worm-like forms.

The functional principles of a hydrostatic skeleton were combined to obtain a physical model which includes geometry, number and length-tension relationships of the elastic elements in the body wall, internal volume and internal pressure. The model skeleton with pre-set internal volume assumes a certain shape and develops a specific internal pressure in order to minimize the potential energy stored in the elastic elements. This shape is calculated as equilibrium state by using finite element methods and optimization techniques. This model is flexible enough to accommodate different geometries and length-tension-relationships of the elastic elements. Presently, the model is implemented with linear length-tension relationships and certain geometrical restrictions, such as uniform width over the entire animal, and rectangular cross sections; the general case is outlined. First simulations with the "unit-worm" yield stable solutions, i.e. stable shapes for all combinations of parameters tested so far. They define the conditions for bringing all muscles to an optimal operating point. We detected a pressure maximum with increasing volume, assessed the contribution of circular muscles to bending, and determined the shapes of animals with different muscle activations in each body half (Chapman-matrix). We summarize our results by the volume rule and stabilization rule, two simple concepts which predict changes in shape as the result of muscle activation.

Animals↗

PubMed1995

Comparison of the effect of monopolar and bipolar cauterization on skeletonized, dissected internal thoracic arteries.

The internal thoracic artery is preferable to the saphenous vein for use as a conduit for coronary artery bypass. More extensive use is possible if this artery is mobilized in a skeletonized form to provide greater length. Internal thoracic arteries are usually mobilized with cauterization. This study compared the effectiveness and effects on neighboring areas of division of the branches of the canine internal thoracic artery with bipolar cauterization and monopolar cauterization. Branch closure was significantly more secure in the bipolar cauterization group, with bleeding in 25 (9%) of 279 branches of 15 internal thoracic arteries treated with monopolar cauterization, in contrast to bleeding in 4 (1.3%) of 306 branches in the 15 internal thoracic arteries treated with bipolar cauterization, which were paired with the group treated with monopolar cauterization. The group of internal thoracic arteries treated with monopolar cauterization had a significantly higher prevalence of leakage when luminal pressure was increased from 120 to 160 mm Hg. Scanning electron microscopy demonstrated partial loss of endothelial cells on the flow surface of internal thoracic arteries treated with bipolar cauterization, compared with almost complete loss of endothelial cells around the orificial areas after monopolar cauterization. Secondary bipolar cauterization treatment caused only slightly more damage than primary treatment, but secondary monopolar cauterization was much more severe and extensive than primary treatment. These data suggest that bipolar cauterization is preferable to monopolar cauterization for skeletonized dissection of the internal thoracic artery.

Animals↗

PubMed1990

The microarchitecture of the intracavernosal smooth muscle and the cavernosal fibrous skeleton.

The bulk of the parenchyma of the corpora cavernosa consists of bundles of smooth muscle fibers. These bundles are oriented in all directions and establish at least 2 insertions upon elements of the fibrous skeleton of the corpora. The fibrous skeleton includes the tunica albuginea and its fibrous columns, the intracavernous fibrous framework, and the periarterial and perineural fibrous sheaths. The cavernous vascular spaces (sinusoids) form an intercommunicating network embedded within the mass of smooth muscle. Thus, the smooth muscle does not form a muscular wall proper for each sinusoid.

Collagen↗

PubMed1998

Forced dissociation of food- and light- entrainable circadian rhythms of rats in a skeleton photoperiod.

To investigate the control over drinking and feeding behavior by the light-entrainable circadian pacemaker, rats were maintained in a 12 h:12 h skeleton photoperiod for 36 days with both food and water restricted to the subjective day. During the restriction period most of the food and water intake was concentrated during the first 4 h of the subjective day. The subsequent release into ad lib conditions under the skeleton photoperiod or continuous dark showed that this increased ingestion during the subjective day persisted for up to 6-10 days. This may indicate the entrainment of a food-entrainable oscillator, which has hitherto been investigated solely in anticipatory activity. The daily activity rhythm returned to its original phase position. Thus the phase of the overt daily rhythm is eventually only determined by the phase of the light-entrainable oscillator (LEO) but can temporarily be influenced by the restriction of food and water access. This restriction apparently stimulated a food-entrainable oscillator in antiphase with the light-entrainable oscillator but failed to phase shift or entrain the light-entrainable oscillator.

Animals↗

PubMed2002

Electron diffraction studies of the calcareous skeletons of bryozoans.

Electron microscopy and electron diffraction were used to investigate mineral crystallites dissociated from the skeletal walls of six species belonging to the Bryozoa, a phylum of predominantly marine colony-forming invertebrate animals. Four cheilostome bryozoans (Flustra foliacea, Membranipora membranacea, Thalamoporella novaehollandiae and Cellarinella foveolata) and two cyclostomes (Fasciculipora ramosa and Hornera robusta) were analysed. In each case, an attempt was made to relate the crystal morphology imaged in situ by scanning electron microscopy with the crystallographic orientation of isolated crystals determined by electron diffraction analysis in the transmission electron microscope. The results showed that the calcitic cheilostome and cyclostome skeletons consisted of closely packed arrays of plate-like Mg-containing calcite crystallites, and that the crystallographic a-axis was preferentially aligned perpendicular to the top and bottom surfaces of the flattened particles. The results suggest that calcite biomineralization occurs under similar crystallographic constraints in the five species studied even though the origins of cheilostomes and cyclostomes are separated by over 300 million years in the fossil record of the bryozoans. Similar studies for the aragonite crystallites in skeletons of M. membranacea indicated that the crystallographic b-axis was preferentially oriented perpendicular to the basal surfaces of irregular plate-like particles.

Animals↗

PubMed2000

Pfsbp1, a Maurer's cleft Plasmodium falciparum protein, is associated with the erythrocyte skeleton.

Antibodies from hyperimmune monkey sera, selected by absorption to Plasmodium falciparum-infected erythrocytes, and elution at acidic pH, allowed us to characterize a novel parasite protein, Pfsbp1 (P. falciparum skeleton binding protein 1). Pfsbp1 is an integral membrane protein of parasite-induced membranous structures associated with the erythrocyte plasma membrane and referred to as Maurer's clefts. The carboxy-terminal domain of Pfsbp1, exposed within the cytoplasm of the host cell, interacts with a 35 kDa erythrocyte skeletal protein and might participate in the binding of the Maurer's clefts to the erythrocyte submembrane skeleton. Antibodies to the carboxy- and amino-terminal domains of Pfsbp1 labelled similar vesicular structures in the cytoplasm of Plasmodium chabaudi and Plasmodium berghei-infected murine erythrocytes, suggesting that the protein is conserved among malaria species, consistent with an important role of Maurer's cleft-like structures in the intraerythrocytic development of malaria parasites.

Amino Acid Sequence↗

PubMed1998

Isolation, characterisation and embryonic expression of WNT11, a gene which maps to 11q13.5 and has possible roles in the development of skeleton, kidney and lung.

The Wnt gene family encodes a set of signalling molecules, thought to play an important role in key processes of embryonic development. In vertebrates as a whole 20 different Wnt genes have been identified to date, however, a complement of only 16 have been identified in man and for some of these the complete coding sequences are unavailable. We have recently isolated the full-length cDNA sequence of a new human WNT gene, WNT11, investigated its genomic organisation and performed detailed expression studies in early human embryos. These have shown that the expression of human WNT11 is restricted to the perichondrium of the developing skeleton, lung mesenchyme, the tips of the ureteric buds and other areas of the urogenital system and the cortex of the adrenal gland. This, for the first time, provides information for the embryonic expression of human WNT11. We have mapped WNT11 to 11q13.5 and this together with its expression in the perichondrium of the developing skeleton, makes it a plausible candidate gene for HBM, which has been previously linked to markers from this region.

Amino Acid Sequence↗

PubMed1999

Parallel and robust skeletonization built on self-organizing elements.

A massively parallel neural architecture is suggested for the approximate computation of the skeleton of a planar shape. Numerical examples demonstrate the robustness of the method. The architecture is constructed from self-organizing elements that allow the extension of the concept of skeletonization to areas remote to image processing.

Journal Article↗

PubMed1993

Natural coral skeleton used as onlay graft for contour augmentation of the face. A preliminary report.

The skeletons of two different genera of Madreporaria were used as onlay graft material for contour augmentation of the face. With coral blocks or granules in a subperiosteal fashion, patients with hemifacial microsomia, maxillary hypoplasia, microgenia, and Treacher Collins' syndrome have been adequately treated. Sixteen augmentations were performed in 14 patients, followed up for 6-34 months. One graft had to be removed because its inadequate fixation resulted in mobility. Delayed healing was observed in another case for the same reason. Coral skeleton is a cheap, natural biomaterial with excellent biocompatibility. It can easily be prepared and shaped, and it maintained its form over the period observed.

Adolescent↗

PubMed2000

Novel potassium channel openers. Part 4: transformation of the 1,4-benzoxazine skeleton into 1,4-benzothiazine, 1,2,3,4-tetrahydroquinoline, 1,2,3,4-tetrahydroquinoxaline, indoline, and 1,5-benzoxazepine.

As part of a search for a new potassium channel opener, the 1,4-benzoxazine skeleton derived from the benzopyran skeleton of cromakalim, was transformed into other fused rings such as 1,4-benzothiazine, 1,2,3,4-tetrahydroquinoline, 1,2,3,4-tetrahydroquinoxaline, indoline, and 1,5-benzoxazepine. The 1,4-benzothiazine derivative displayed approximately 20 times more potent vasorelaxant activity than cromakalim.

Animals↗

PubMed1988

Influence of calmodulin on the human red cell membrane skeleton.

The calcium receptor calmodulin interacts with components of the human red cell membrane skeleton as well as with the membrane. Under physiological salt conditions, calmodulin has a calcium-dependent affinity for spectrin, one of the major components of the membrane skeleton. It is apparent from our results that calmodulin inhibits the ability of erythrocyte spectrin (when preincubated with filamentous actin) to create nucleation centers and thereby to seed actin polymerization. The gelation of filamentous actin induced by spectrin tetramers is also inhibited by calmodulin. The inhibition is calcium dependent and decreases with increasing pH, similar to the binding of calmodulin to spectrin. Direct binding studies using aqueous two-phase partition indicate that calmodulin interferes with the binding of actin to spectrin. Even in the presence of protein 4.1, which is believed to stabilize the ternary complex, calmodulin has an inhibitory effect. Since calmodulin also inhibits the corresponding activities of brain spectrin (fodrin), it appears likely that calmodulin may modulate the organization of cytoskeletons containing actin and spectrin or spectrin analogues.

Actins↗

PubMed2006

Effect of surface composition on electronic structure, stability, and electrocatalytic properties of Pt-transition metal alloys: Pt-skin versus Pt-skeleton surfaces.

The surface properties of PtM (M = Co, Ni, Fe) polycrystalline alloys are studied by utilizing Auger electron spectroscopy, low energy ion scattering spectroscopy, and ultraviolet photoemission spectroscopy. For each alloy initial surface characterization was done in an ultrahigh vacuum (UHV) system, and depending on preparation procedure it was possible to form surfaces with two different compositions. Due to surface segregation thermodynamics, annealed alloy surfaces form the outermost Pt-skin surface layer, which consists only platinum atoms, while the sputtered surfaces have the bulk ratio of alloying components. The measured valence band density of state spectra clearly shows the differences in electronic structures between Pt-skin and sputtered surfaces. Well-defined surfaces were hereafter transferred out from UHV and exposed to the acidic (electro)chemical environment. The electrochemical and post-electrochemical UHV surface characterizations revealed that Pt-skin surfaces are stable during and after immersion to an electrolyte. In contrast all sputtered surfaces formed Pt-skeleton outermost layers due to dissolution of transition metal atoms. Therefore, these three different near-surface compositions (Pt-skin, Pt-skeleton, and pure polycrystalline Pt) all having pure-Pt outermost layers are found to have different electronic structures, which originates from different arrangements of subsurface atoms of the alloying component. Modification in Pt electronic properties alters adsorption/catalytic properties of the corresponding bimetallic alloy. The most active systems for the electrochemical oxygen reduction reaction are established to be the Pt-skin near-surface composition, which also have the most shifted metallic d-band center position versus Fermi level.

Journal Article↗

PubMed2002

Intact carbohydrate structures as part of the melanoidin skeleton.

Model melanoidins from monomeric, oligomeric, and polymeric carbohydrates, and amino acids formed under aqueous as well as water-free reaction conditions, were submitted to acidic catalyzed hydrolysis. Their degradation products were detected qualitatively and quantitatively by HPTLC and HPLC-DAD. A considerable amount of monomer carbohydrates from hydrolysis of model melanoidins formed under water-free reaction conditions was detected. It can be seen clearly that the amount of carbohydrates released increased with increasing degree of polymerization of the carbohydrates used as starting material. In comparison, the hydrolysis of melanoidins formed in aqueous condition resulted in only a small glucose release. It seems that in the Maillard reaction under water-free conditions, a significant amount of di- and oligomer carbohydrates were incorporated into the melanoidin skeleton as complete oligomer with intact glycosidic bond, forming side chains at the melanoidin skeleton. Additional side chains could be formed by transglycosylation reactions. With increasing water content, hydrothermolytic as well as retro-aldol reactions of the starting carbonyl components became significant, and therefore the possibility of forming side chains decreased. The results are consistent with the postulated melanoidin structure being built up mainly from sugar degradation products, probably branched via amino compounds.

Alanine↗