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At least 199 records · Page 11Linked to original sources

PubMed1999

Effect of skeletonization of the internal thoracic artery on vessel wall integrity.

BACKGROUND: This study was conceived to evaluate the effect of internal thoracic artery (ITA) skeletonization on vessel wall integrity. METHODS: Forty consecutive patients undergoing coronary artery bypass were randomized to receive a skeletonized (n = 22) or a pedicled (n = 18) ITA graft. ITA harvesting was performed by 2 experienced surgeons using the same instrumentation and technique. Specimens were examined by light and electron microscope in order to assess vascular wall integrity. A specific immunohistochemical staining and a computerized method were used to quantify the degree of endothelial integrity after surgical preparation. RESULTS: Morphologic analysis revealed 2 cases of limited subadventitial hemorrhage (one for each group) and no case of major arterial damage. Immunohistochemical staining demonstrated an extremely high degree of maintenance of the endothelial integrity in both groups (97.2% +/- 1.9% in the skeletonized and 96.8% +/- 2.1% in the pedicled one; p = 0.53). CONCLUSIONS: Skeletonization does not affect ITA wall integrity in humans submitted to coronary artery bypass procedures.

Aged↗

PubMed2001

Direct measurement of the area expansion and shear moduli of the human red blood cell membrane skeleton.

The area expansion and the shear moduli of the free spectrin skeleton, freshly extracted from the membrane of a human red blood cell (RBC), are measured by using optical tweezers micromanipulation. An RBC is trapped by three silica beads bound to its membrane. After extraction, the skeleton is deformed by applying calibrated forces to the beads. The area expansion modulus K(C) and shear modulus mu(C) of the two-dimensional spectrin network are inferred from the deformations measured as functions of the applied stress. In low hypotonic buffer (25 mOsm/kg), one finds K(C) = 4.8 +/- 2.7 microN/m, mu(C) = 2.4 +/- 0.7 microN/m, and K(C)/mu(C) = 1.9 +/- 1.0. In isotonic buffer, one measures higher values for K(C), mu(C), and K(C)/mu(C), partly because the skeleton collapses in a high-ionic-strength environment. Some data concerning the time evolution of the mechanical properties of the skeleton after extraction and the influence of ATP are also reported. In the Discussion, it is shown that the measured values are consistent with estimates deduced from experiments carried out on the intact membrane and agree with theoretical and numerical predictions concerning two-dimensional networks of entropic springs.

Adenosine Triphosphate↗

PubMed1997

Influence of network topology on the elasticity of the red blood cell membrane skeleton.

A finite-element network model is used to investigate the influence of the topology of the red blood cell membrane skeleton on its macroscopic mechanical properties. Network topology is characterized by the number of spectrin oligomers per actin junction (phi a) and the number of spectrin dimers per self-association junction (phi s). If it is assumed that all associated spectrin is in tetrameric form, with six tetramers per actin junction (i.e., phi a = 6.0 and phi s = 2.0), then the topology of the skeleton may be modeled by a random Delaunay triangular network. Recent images of the RBC membrane skeleton suggest that the values for these topological parameters are in the range of 4.2 < phi a < 5.5 and 2.1 < phi s < 2.3. Model networks that simulate these realistic topologies exhibit values of the shear modulus that vary by more than an order of magnitude relative to triangular networks. This indicates that networks with relatively sparse nontriangular topologies may be needed to model the RBC membrane skeleton accurately. The model is also used to simulate skeletal alterations associated with hereditary spherocytosis and Southeast Asian ovalocytosis.

Actins↗

PubMed1998

Structure of the erythrocyte membrane skeleton as observed by atomic force microscopy.

The structure of the membrane skeleton on the cytoplasmic surface of the erythrocyte plasma membrane was observed in dried human erythrocyte ghosts by atomic force microscopy (AFM), taking advantage of its high sensitivity to small height variations in surfaces. The majority of the membrane skeleton can be imaged, even on the extracellular surface of the membrane. Various fixation and drying methods were examined for preparation of ghost membrane samples for AFM observation, and it was found that freeze-drying (freezing by rapid immersion in a cryogen) of unfixed specimens was a fast and simple way to obtain consistently good results for observation without removing the membrane or extending the membrane skeleton. Observation of the membrane skeleton at the external surface of the cell was possible mainly because the bilayer portion of the membrane sank into the cell during the drying process. The average mesh size of the spectrin network observed at the extracellular and cytoplasmic surfaces of the plasma membrane was 4800 and 3000 nm2, respectively, which indicates that spectrin forms a three-dimensionally folded meshwork, and that 80% of spectrin can be observed at the extracellular surface of the plasma membrane.

Cell Fractionation↗

PubMed1987

Model of red blood cell membrane skeleton: electrical and mechanical properties.

A theoretical membrane skeleton model of erythrocyte has been developed and successfully applied to interpret electrical and mechanical properties of the red blood cell spectrin-actin network. The model is based on the structure of the membrane skeleton that is comprised of unit cells each containing an actin protofilament and shooting forth a few spectrin heterodimers. The loose ends of the heterodimers of adjacent cells can form bonds with each other giving rise to an integrated network. The number of bonds depends on the temperature. The bond length being excessive (2.6 times the distance between the centers of adjacent cells), the bonds are flexible, and can thus be regarded as entropy springs. The advanced model has been employed to calculate the shear modulus of the membrane skeleton as well as to establish its temperature dependence. In a wide range of temperatures mu(T) is a decreasing function well fitting the experimental data. The relationship between the membrane bilayer-free size of the skeleton and the ionic strength of the solution has been derived to appear in good agreement with the results obtained previously. Experimental data combined with the advanced theory yield the average number of heterodimers per unit cell, m0, as equal to ca. 5; the spectrin heterodimer charge has been estimated.

Actins↗

PubMed1996

Anion exchanger 1 (band 3) is required to prevent erythrocyte membrane surface loss but not to form the membrane skeleton.

The red blood cell (RBC) membrane protein AE1 provides high affinity binding sites for the membrane skeleton, a structure critical to RBC integrity. AE1 biosynthesis is postulated to be required for terminal erythropoiesis and membrane skeleton assembly. We used targeted mutagenesis to assess AE1 function in vivo. RBCs lacking AE1 spontaneously shed membrane vesicles and tubules, leading to severe spherocytosis and hemolysis, but the levels of the major skeleton components, the synthesis of spectrin in mutant erythroblasts, and skeletal architecture are normal or nearly normal. The results indicate that AE1 does not regulate RBC membrane skeleton assembly in vivo but is essential for membrane stability. We postulate that stabilization is achieved through AE1-lipid interactions and that loss of these interactions is a key pathogenic event in hereditary spherocytosis.

Animals↗

PubMed2000

Adiabatic compressibility of red blood cell membrane: influence of skeleton.

Measurements of ultrasound velocity and density were used for determination of the adiabatic compressibility of red blood cells (RBC) during detachment of the membrane skeleton. Skeleton detachment was induced by addition of nystatin into a low ionic strength RBC suspension resulting in an increase (10%) of the ultrasound velocity concentration increment, [u], while the specific volume of cells, phi(v) did not change significantly. Changes of the concentration increment had rather long kinetics and were not completed even after 60 min. Both [u] and phiV values were used for calculation of the specific apparent adiabatic compressibility of RBC, phiK/beta0. The value of the specific apparent compressibility decreases following addition of nystatin. This corresponds to an increase in the volume elastic modulus of RBC membranes during detachment of the membrane skeleton. Control experiments with large unilamellar liposomes at conditions similar to that performed with the RBC did not reveal significant changes of [u] after the addition of nystatin. Our results show that the role of the membrane skeleton probably consists in maintaining higher compressibility of the RBC membranes. This may partly provide conditions for conformational changes of RBC membrane proteins.

Cell Size↗

PubMed2002

Comparative diagnostic value and therapeutic relevance of magnetic resonance imaging and bone marrow scintigraphy in patients with metastatic solid tumors of the axial skeleton.

PURPOSE: To evaluate the comparative impact of magnetic resonance imaging (MRI) and bone marrow scintigraphy (BMS) in bone marrow metastases of solid tumors. METHODS: In 20 patients with solid tumors MRI of the axial skeleton and whole-body BMS were retrospectively reviewed. Detectability of metastases, extent of disease and therapeutic implications were assessed. RESULTS: In 15/20 (75%) patients MRI and BMS concordantly revealed bone marrow metastases of the axial skeleton. In nine of these 15 patients (60%) MRI showed more metastases. Local radiotherapy or surgery was performed in seven of these cases (78%). BMS detected additional metastases of the appendicular skeleton in 8/15 (53%) patients. In 4/20 cases (20%) the imaging findings were discordant. In three patients with degenerative changes (n=2) or lipoma (n=1) BMS was false positive. In another patient BMS failed to detect metastases proven by MRI and clinical follow-up resulting in subsequent radiation therapy. One patient had normal bone marrow. CONCLUSION: MRI appears to be more sensitive and specific in the detection of bone marrow metastases in the axial skeleton and is of clinical importance for subsequent local therapy.

Bone Marrow Neoplasms↗

PubMed2003

Estrogen deposits extra mineral into bones of female rats in puberty, but simultaneously seems to suppress the responsiveness of female skeleton to mechanical loading.

To first test the possible effect of gender on the responsiveness of growing rat skeleton to mechanical loading, 5-week-old littermates of 25 male and 25 female rats were subjected to either free-cage activity or treadmill training for a period of 14 weeks (experiment 1). Using peripheral quantitative computed tomography (pQCT) and mechanical testing of the femoral neck, we observed female rats exhibiting a clearly lower responsiveness to external loading than male rats (+3.0% vs +25% in cross-sectional area (CSA), +4.2% vs +27% in the bone mineral content (BMC), -0.6% vs +10% in volumetric bone mineral density (BMD), and +4.7% vs +28% in fracture strength (F(max)) of the femoral neck). Also, relative to the mechanical demands placed on the skeleton, the bones of the young female rats were considerably denser (>50%) than those of the males. In the subsequent experiment 2, we repeated the above-noted first experiment with 33-week-old rats and observed virtually identical exercise-induced benefits (+2.1% vs +10% in CSA, +3.4% vs +18% in BMC, +2.5% vs +23% in BMD, and -1.1% vs +27% in F(max) in females vs males, respectively) and the growth/puberty-related condensation of mineral into female bones. Finally, in experiment 3, 60 littermates of 3-week-old female rats were first subjected to sham operation or ovariectomy and then further randomized to exercise or control groups, respectively, to study whether the condensation of mineral into female bones and their lower responsiveness to loading were attributable to the effects of estrogen. At the end of the 16-week intervention, our pQCT and mechanical testing analysis showed not only the anticipated effect of reduced bone density in the ovariectomized rats ( approximately -20%) but also the hypothesized better responsiveness to mechanical loading in these estrogen-depleted rats (-3.5% vs +9.1% in CSA, -0.4% vs +12% in BMC, +4.4% vs +9.6% in BMD, and -4.2% vs +16% in F(max) in SHAM vs OVX, respectively). In conclusion, the results of our series of three experiments suggest that as such estrogen seems to have very little primary effect on the sensitivity of female bone to respond to external loading, but rather deposits extra stock of mineral into female bones in puberty. This estrogen-driven extra condensation of the female skeleton seems to persist into adulthood, simultaneously damping the responsiveness of the female skeleton to mechanical loading.

Animals↗

PubMed2005

Syntheses and structures of hypervalent pentacoordinate carbon and boron compounds bearing an anthracene skeleton--elucidation of hypervalent interaction based on X-ray analysis and DFT calculation.

Pentacoordinate and tetracoordinate carbon and boron compounds (27, 38, 50-52, 56-61) bearing an anthracene skeleton with two oxygen or nitrogen atoms at the 1,8-positions were synthesized by the use of four newly synthesized tridentate ligand precursors. Several carbon and boron compounds were characterized by X-ray crystallographic analysis, showing that compounds 27, 56-59 bearing an oxygen-donating anthracene skeleton had a trigonal bipyramidal (TBP) pentacoordinate structure with relatively long apical distances (ca. 2.38-2.46 A). Despite the relatively long apical distances, DFT calculation of carbon species 27 and boron species 56 and experimental accurate X-ray electron density distribution analysis of 56 supported the existence of the apical hypervalent bond even though the nature of the hypervalent interaction between the central carbon (or boron) and the donating oxygen atom was relatively weak and ionic. On the other hand, X-ray analysis of compounds 50-52 bearing a nitrogen-donating anthracene skeleton showed unsymmetrical tetracoordinate carbon or boron atom with coordination by only one of the two nitrogen-donating groups. It is interesting to note that, with an oxygen-donating skeleton, the compound 61 having two chlorine atoms on the central boron atom showed a tetracoordinate structure, although the corresponding compound 60 with two fluorine atoms showed a pentacoordinate structure. The B-O distances (av 2.29 A) in 60 were relatively short in comparison with those (av 2.44 A) in 59 having two methoxy groups on the central boron atom, indicating that the B-O interaction became stronger due to the electron-withdrawing nature of the fluorine atoms.

Journal Article↗

PubMed1997

Antitumor agents. 180. Chemical Studies and cytotoxic evaluation of cumingianosides and cumindysoside A, antileukemic triterpene glucosides with a 14,18-cycloapotirucallane skeleton.

Treatment of cumingianosides and cumindysoside A, which possess a 14,18-cycloapotirucallane skeleton, with p-toluenesulfonic acid in CH2Cl2 yielded new triterpene glucosides. Cumingianoside A (1) gave 10 and 11, along with cumingianoside Q (5). The structures of 10 and 11 were determined on the basis of spectral examination and contained a dammar-13(17)-ene and a 17(R),23(R)-epoxydammarane skeleton, respectively. Cumingianoside C (2) afforded, together with cumingianoside P (6), products 12 and 13, which were similar to 10 and 11, respectively. With a short reaction time at room temperature, cumingianoside E (3) yielded cumingianoside D (4). In contrast, when 3 was treated with p-toluenesulfonic acid in CH2Cl2 overnight at 5 degrees C, it gave two products, 9 and 14. Extensive spectroscopic examination revealed that 9 possessed a dammar-12-ene skeleton, while 14 was a pentacyclic tetranortriterpene glucoside with a novel skeleton. Cumindysoside A (8) gave a product (15) similar to 14. The cytotoxicities of 9-15 were evaluated against a panel of 58 human tumor cell lines. Compounds 11-15 exhibited potent cytotoxicity with log GI50 values ranging from -7.11 to -4.94, especially against leukemia and colon-tumor cell lines.

Antineoplastic Agents, Phytogenic↗

PubMed2002

Importance of the methyl group at C10 of squalene for hopene biosynthesis and novel carbocyclic skeletons with 6/5 + 5/5 + (6) ring system(s).

[reaction: see text] Incubation of (6E,10E,14E,18E)-2,6,10,19,23-pentamethyl-tetracosa-2,6,10,14,18,22-hexaene with Alicyclobacillus acidocaldarius hopene cyclase afforded four products having two types of carbocyclic skeletons, i.e., two hopane products and two products having an unprecedented carbocyclic skeleton of 6/5 + 5/5 +6 pentacyclic and 6/5 + 5/5 tetracyclic ring systems. The former two hopane skeletons were formed from the bioconversion of C15-desmethylsqualene and the latter two skeletons from that of C10-desmethylsqualene.

Bacillus↗

PubMed1995

The polarity of the membrane skeleton in retinal pigment epithelial cells of developing chicken embryos and in primary culture.

We studied the morphogenesis and the membrane skeleton in the retinal pigment epithelium during chicken embryogenesis and in culture, by using immunofluorescence and electron microscopy. During embryogenesis two distinct membrane skeletal structures were formed, an apical and a basolateral one. The former was seen in the apical surface already in the 10-day-old embryos. It was comprised of ankyrin and alpha-fodrin and showed a codistribution with Na+,K(+)-ATPase and an as yet uncharacterized cadherin-like molecule. The basolateral membrane skeleton was seen in the lateral walls already in the 10-day-old embryos, and later, between the 13th and 17th embryonic days, it also appeared at the basal membrane, coincidentally with the formation of the basal infoldings. It consisted of ankyrin and alpha-fodrin, but did not codistribute with any of the integral membrane proteins studied (Na+,K(+)-ATPase and cadherins). In culture, the retinal pigment epithelial cells retained their polarized morphology. Compared with the situation in vivo, however, there was a distinct translocation of the membrane skeletal components fodrin and ankyrin from the apical surface to the lateral walls, accompanied by a similar redistribution of Na+,K(+)-ATPase and the cadherin-like molecule. The results suggest that (1) there is, in the retinal pigment epithelium, an apical Na+,K(+)-ATPase-membrane skeleton structure stabilized by contacts between the retinal pigment epithelium and the neural retina, possibly mediated by a cadherin-like molecule, and that (2) there is another fodrin/ankyrin-based membrane skeleton in the basolateral walls that is important for the maintenance of the extensive folding of these surface areas.

Animals↗

PubMed2001

Desmocytes in the calicoblastic epithelium of the stony coral Mycetophyllia reesi and their attachment to the skeleton.

Desmocytes scattered over the surface of the corallum of the scleractinian Mycetophyllia reesi attach the calicoblastic tissue to the skeleton. The structure of the desmocyte is generally consistent with that of other scleractinians except for their more rectangular profiles and greater size. However, the extent of attachment is distinctive, and the mode of attachment to mineral is described for the first time. The skeleton contains dual rows of interconnected pits between the septa, within and among which desmocytes form virtually uninterrupted sheets. Desmocytes terminate with hemidesmosomes that attach the epithelium to a fibrillar basal lamina. Fibrils extend from the basal lamina into the skeletal matrix anchoring tissue firmly to the skeleton. In addition, the basal lamina itself appears to be incorporated within the organic matrix during growth, partitioning the skeleton into compartments. Because the skeletal organic matrix is physicochemically labile during demineralization, these intraskeletal details cannot be observed unless polycationic dyes such as Ruthenium red or other glycan precipitating agents are employed in the fixative sequence.

Animals↗

PubMed1984

[Radiologic studies of the correlation between the mineral content of the peripheral and axial skeleton].

Estimations of mineral content of normal human skeletons using three radiological methods have led to the following conclusions: 1. Bone density of vertebral bodies increases from T.6 to L.5. Average values for thoracic and lumbar vertebrae are 0.71 and 0.88 g./cm2. 2. There is no linear correlation between the mineral content of an easily accessible peripheral bone and of the vertebrae. 3. The method for bone mineral estimation, using thulium170, which was developed in Dresden, can be used for the peripheral and central skeleton. 4. 125-photon absorption us useful for measurements on the peripheral skeleton, where it is the most elegant and accurate method. It is, however, useless for measurements of the central skeleton.

Adolescent↗

PubMed2001

Bilateral skeletonized internal thoracic artery grafts in patients with diabetes mellitus.

OBJECTIVE: Increased risk of deep sternal infections has prohibited routine bilateral internal thoracic artery grafting in diabetic patients. The technique for harvesting the skeletonized internal thoracic artery provides the potential to minimize this risk. The purpose of this study was to compare the outcome of bypass grafting with bilateral skeletonized internal thoracic arteries in diabetic and nondiabetic patients. METHODS: From May 1996 to April 1998, 231 consecutive diabetic and 534 nondiabetic patients underwent bilateral skeletonized internal thoracic artery grafting. Mean age was 66 years. Compared with the nondiabetic group, the diabetic group comprised more women (29% vs 18%, P =.001), had a greater prevalence of hypertension (53% vs 44%, P =.019) and congestive heart failure (20% vs 14%, P =.016), but a lower prevalence of preoperative acute myocardial infarction (26% vs 34%, P =.027). RESULTS: Operative mortality of diabetic patients was comparable with that of nondiabetic patients (3% vs 2.6%). The two groups also had similar occurrences of deep sternal infection (2.6% vs 1.7%, respectively, P =.40). Deep sternal infection was significantly more prevalent in obese, diabetic women (3/20 = 15%) than in diabetic patients without this combination of risk factors (3/211 = 1.4%, P <.0001) (odds ratio 11.1, confidence interval 2.1-59.4). Diabetic patients also had a higher incidence of stroke (3.5% vs 0.9%, P =.014). Three-year actuarial survival of diabetic patients was lower (91.3% vs 94.7%, P =.083). CONCLUSIONS: Bilateral skeletonized internal thoracic artery grafting is a good surgical revascularization option in diabetic patients. Operative mortality and prevalence of sternal infection are comparable with those of nondiabetic patients. However, the risk of sternal infection in obese diabetic women is high, and for them we advocate the use of a single artery instead of bilateral internal thoracic arteries.

Aged↗

PubMed1981

Elliptical erythrocyte membrane skeletons and heat-sensitive spectrin in hereditary elliptocytosis.

Erythrocyte membranes (ghosts) and membrane skeletons (submembranous reticula of spectrin, actin, and protein 4.1 prepared by extracting ghosts with Triton X-100) from 15 patients with hereditary elliptocytosis (HE) were elliptical, which indicates that the primary defect responsible for the abnormal shape of these cells resides in the skeleton. The protein composition of HE skeletons was normal, but in three kindreds purified spectrin heterodimer from 7/7 HE patients was heat sensitive and denatured at 48.0 +/- 0.1 degrees C instead of 49.0 +/- 0.3 degrees C (P less than 0.0005). Heat sensitivity was detected by precipitation and, in the spectrin from one patient, by changes in circular dichroism. In one other kindred spectrin dimer from 3/3 patients denatured at the normal temperature. In two of the three kindreds with heat-sensitive spectrin, intact erythrocytes exhibited budding and fragmentation at the temperature at which spectrin denatured. In the third kindred spectrin was heat sensitive, but erythrocytes were not. The symptoms in the latter kindred were clinically more severe (hemolytic HE with spherocytosis) than in the other three (mild HE). We conclude that defects in the erythrocyte membrane skeleton may be a common feature of HE. As judged by heat denaturation of erythrocytes and purified spectrin dimer, three phenotypically distinct forms of HE exist, two of which are characterized by defective, heat-sensitive spectrin. It remains to be determined whether the molecular defect in spectrin responsible for heat sensitivity is the primary genetic defect responsible for HE.

Drug Stability↗

PubMed2002

Proteomic analysis of a detergent-resistant membrane skeleton from neutrophil plasma membranes.

Plasma membranes are organized into functional domains both by liquid-ordered packing into "lipid rafts," structures that resist Triton extraction, and by attachments to underlying cytoskeletal proteins in assemblies called "membrane skeletons." Although the actin cytoskeleton is implicated in many lipid raft-mediated signaling processes, little is known about the biochemical basis for actin involvement. We show here that a subset of plasma membrane skeleton proteins from bovine neutrophils co-isolates with cholesterol-rich, detergent-resistant membrane fragments (DRMs) that exhibit a relatively high buoyant density in sucrose (DRM-H; d approximately 1.16 g/ml). By using matrix-assisted laser desorption/ionization time of flight and tandem mass spectrometry, we identified 19 major DRM-H proteins. Membrane skeleton proteins include fodrin (nonerythroid spectrin), myosin-IIA, myosin-IG, alpha-actinin 1, alpha-actinin 4, vimentin, and the F-actin-binding protein, supervillin. Other DRM-H components include lipid raft-associated integral membrane proteins (stomatin, flotillin 1, and flotillin 2), extracellular surface-bound and glycophosphatidylinositol-anchored proteins (IgM, membrane-type 6 matrix metalloproteinase), and intracellular dually acylated signaling proteins (Lyn kinase, Galpha(i-2)). Consistent with cytoskeletal association, most DRM-H-associated flotillin 2, Lyn, and Galpha(i-2) also resist extraction with 0.1 m octyl glucoside. Supervillin, myosin-IG, and myosin-IIA resist extraction with 0.1 m sodium carbonate, a treatment that removes all detectable actin, suggesting that these cytoskeletal proteins are proximal to the DRM-H bilayer. Binding of supervillin to the DRM-H fragments is confirmed by co-immunoaffinity purification. In spreading neutrophils, supervillin localizes with F-actin in cell extensions and in discrete basal puncta that partially overlap with Galpha(i) staining. We suggest that the DRM-H fraction represents a membrane skeleton-associated subset of leukocyte signaling domains.

Actins↗