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PubMed1996

Membrane skeleton involvement in cell fusion kinetics: a parameter that correlates with erythrocyte osmotic fragility.

Spectrin levels in erythrocytes have been related to several biomechanical and biophysical membrane properties essential to the survival and function of the cell. Populations of erythrocytes display a natural and finite range of sensitivities to osmotic shock that has been directly correlated, in studies from other laboratories, to the presence of spectrin. We used a procedure to isolate subpopulations of 1) the osmotically most sensitive and 2) the osmotically most resistant erythrocyte membranes in an attempt to select for membranes enriched and depleted in spectrin (and/or a related component). The mechanical function of the spectrin-based membrane skeleton was further explored in these two subpopulations by searching for any effect on the time-dependent increase in fusion zone diameter in pairs of electrofused erythrocyte ghosts as a model for cell fusion. The results clearly show that the diameter expansions in fusions of membranes from osmotically resistant erythrocytes are faster in the early stage (up to 9 to 10 s after fusion) but do not thereafter expand as far as in fusions of membranes from osmotically sensitive membranes.

Animals↗
PubMed1996

An elastic network model based on the structure of the red blood cell membrane skeleton.

A finite element network model has been developed to predict the macroscopic elastic shear modulus and the area expansion modulus of the red blood cell (RBC) membrane skeleton on the basis of its microstructure. The topological organization of connections between spectrin molecules is represented by the edges of a random Delaunay triangulation, and the elasticity of an individual spectrin molecule is represented by the spring constant, K, for a linear spring element. The model network is subjected to deformations by prescribing nodal displacements on the boundary. The positions of internal nodes are computed by the finite element program. The average response of the network is used to compute the shear modulus (mu) and area expansion modulus (kappa) for the corresponding effective continuum. For networks with a moderate degree of randomness, this model predicts mu/K = 0.45 and kappa/K = 0.90 in small deformations. These results are consistent with previous computational models and experimental estimates of the ratio mu/kappa. This model also predicts that the elastic moduli vary by 20% or more in networks with varying degrees of randomness. In large deformations, mu increases as a cubic function of the extension ratio lambda 1, with mu/K = 0.62 when lambda 1 = 1.5.

Biophysical Phenomena↗
PubMed2003

Reconstructive options in the treatment of osteoradionecrosis of the craniomaxillofacial skeleton.

Osteoradionecrosis (ORN) of the craniomaxillofacial skeleton is a serious and debilitating complication that can occur following radiation therapy in the head and neck. Patients require effective treatment, which eradicates diseased tissue and restores function with minimal additional morbidity in a single stage, a requirement fulfilled in many cases by free tissue transfer. In a 6-year period from 1994 to 1999, 21 patients with ORN were treated by wide resection and free-flap reconstruction. The median interval between radiation therapy and ORN was 4 years (range: 1-33 years). The median radiation dose was 6000cGy. The affected areas were the mandible (15 patients), the temporal bone (three patients), the maxilla (one patient), the cervical vertebrae (one patient) and the frontal bone (one patient). Clinical symptoms included pain, ulceration, a persistent draining fistula, exposure of bone or hardware, and pathological fracture or non-union of bone. Six patients had had previous unsuccessful attempts at conservative surgical resection. Ten patients had preoperative hyperbaric oxygen (HBO) therapy. A number of different flaps were used for reconstruction in these patients. These included free fibula flaps (13 patients), iliac crest flaps (two patients), scapula flaps (three patients) and rectus abdominis flaps (three patients). All patients achieved relief from their presenting symptoms and primary bone or wound healing. One flap (4.8%) was lost. This was successfully reconstructed in a subsequent procedure. There were three flap re-explorations, two for arterial thrombosis and one for venous thrombosis. Conservative measures, such as limited debridement and HBO therapy, may be effective in preventing the progression of ORN. However, they fail to eradicate established ORN, which requires radical surgical resection followed by functional reconstruction with well-vascularised tissue.

Aged↗
PubMed2001

Transition metals and carbohydrates: the methyl-4,6-O-benzylidene-2,3-diazo-2,3-dideoxy-alpha-D-mannopyranoside skeleton as building block for new chiral nitrogen chelates.

This paper describes a straightforward strategy for the synthesis of new nitrogen chelates by employing the chirality of readily available sugars. Thus, diimino and diamino ligands can be attained easily by using the methyl-4,6-O-benzylidene-2,3-diazo-2,3-dideoxy-alpha-D-mannopyranoside skeleton. The coordinating ability of one ligand has also been assessed by preparing a square-planar palladium complex [PdCl2(N,N-chelate)].

Ligands↗
PubMed2003

Vitamin A deficiency in the late gastrula stage rat embryo results in a one to two vertebral anteriorization that extends throughout the axial skeleton.

Vitamin A and its metabolites are known to be involved in patterning the vertebrate embryo. Study of the effect of vitamin A on axial skeletal patterning has been hindered by the fact that deficient embryos do not survive past midgestation. In this study, pregnant vitamin A-deficient rats were maintained on a purified diet containing limiting amounts of all-trans retinoic acid (12 microg atRA/g diet) and given a daily oral bolus dose of retinol starting at embryonic day 0.5, 8.25, 8.5, 8.75, 9.25, 9.5, 9.75, or 10.5. Embryos were recovered at E21.5 for analysis of the skeleton and at earlier times for analysis of select mRNAs. Normal axial skeletal development and patterning were observed in embryos from pregnant animals receiving retinol starting on or before E8.75. Delay of retinol supplementation to E9.5 or later resulted in a marked increase in both occurrence and severity of skeletal malformations, extending from the craniocervical to sacral regions. Embryos from the groups receiving retinol starting at E9.5 and E9.75 had one-vertebral anterior transformations of the cervical, thoracic, lumbar, and sacral vertebrae. Few embryos survived in the E10.5 group, but these embryos yielded the most severe and extensive anteriorization events. The skeletal alterations seen in vitamin A deficiency are associated with posterior shifts in the mesodermal expression of Hoxa-4, Hoxb-3, Hoxd-3, Hoxd-4, and Hoxa-9 mRNAs, whereas the anterior domains of Hoxb-4 and Cdx2 expression are unaltered. This work defines a critical window of development in the late gastrula-stage embryo when vitamin A is essential for normal axial skeletal patterning and shows that vitamin A deficiency causes anterior homeotic transformations extending from the cervical to lumbosacral regions.

Animals↗
PubMed1998

The role and source of 5'-deoxyadenosyl radical in a carbon skeleton rearrangement catalyzed by a plant enzyme.

The last step in the biosynthesis of tropane alkaloids is the carbon skeleton rearrangement of littorine to hyoscyamine. The reaction is catalyzed by a cell-free extract prepared from cultured hairy roots of Datura stramonium. Adenosylmethionine stimulated the rearrangement 10-20-fold and showed saturation kinetics with an apparent Km of 25 microM. It is proposed that S-adenosylmethionine is the source of a 5'-deoxyadenosyl radical which initiates the rearrangement in a similar manner as it does in analogous rearrangements catalyzed by coenzyme B12-dependent enzymes. Possible roles of S-adenosylmethionine as a radical source in higher plants are discussed.

Atropine↗
PubMed1992

Gonadotropin-releasing hormone agonists and the skeleton.

OBJECTIVE: To review and evaluate published studies that have assessed the effect on the skeleton of treatment with gonadotropin-releasing hormone agonists (GnRH-a). The effects on bone biochemistry and bone density are presented. DATA IDENTIFICATION: Major studies reporting on bone and GnRH-a use were identified through Excerpta Medica and Medline searches. STUDY SELECTION: Those that have reported on GnRH-a treatment in patients where data relating to skeletal metabolism or bone density were obtained. RESULTS: Changes in bone biochemistry during GnRH-a therapy indicate that there is altered skeletal metabolism associated with GnRH-a use. The data on changes in bone density after a 6-month course of GnRH-a therapy generally indicate some bone loss. The magnitude of any detected change in bone density depends on the skeletal site assessed and the method of assessment. No significant bone loss has been documented in cortical bone; any bone loss occurring is usually associated with trabecular bone and is partially or completely reversible on withdrawal of GnRH-a treatment. CONCLUSIONS: A 6-month course of GnRH-a therapy may result in a small reduction in trabecular bone density that is partially or completely reversible on withdrawal of treatment. The magnitude of any persistent residual bone loss is unlikely to be of clinical relevance and the benefits of a single 6-month course of GnRH-a therapy should not be withheld on the basis of a possible effect on bone.

Bone Density↗
PubMed1979

Rapid determination of DDT and related compounds in soils via carbon skeleton gas chromatography-mass spectrometry.

The levels of DDT and related compounds in soil samples from an apple orchard have been determined. Extraction of residues was achieved by cyclic steam extraction and by conventional solvent extraction methods allowing comparison of the two methods. Determination of the organochlorine residues present in the extracts was performed by gas-liquid chromatography-electron-capture detection and by carbon skeleton gas chromatograph-mass spectrometry. Excellent agreement between the two determination techniques was achieved. Prior to application of p,p'-DDT, a value of approximately 0.4 ppm was obtained for Delta DDT in the soil. The major component of this total was DDE. After spraying the apple trees with technical grade, p,p'-DDT, Delta DDT for the soil rose to 0.6 ppm. This increase was due to translocation of p,p'-DDT from the trees.

DDT↗
PubMed2001

Study of Croatian non-aromatic naphthalane constituents with skeletons analogous to bioactive compounds.

Non-aromatic Croatian naphthalane (NAN), shown to be efficace in the treatment of psoriasis vulgaris, was studied in order to improve our understanding of its constituents, which may be potentially responsible for its bioactivity. The components steranes and hopanes were analysed. Since NAN is a complex mixture of hydrocarbons, high-resolution GC and GC-MS were applied as the methods of choice in the study. The GC chromatogram of NAN showed a remarkable cluster in which sterane peaks prevail and composed 33+/- 1% of the sample. Identified steranes (by GC-MS) represented almost half of the cluster (48%). They were in the range from norcholestanes up to propyl cholestanes. The amount of alpha-steranes (8.9%) was higher than of beta-steranes (6.4%) and regular steranes (15.3%) dominated in the ratio 17:1 over the rearranged ones (0.9%). Steranes conserved the skeleton of bio-precursors and remained analogues of bioactive compounds, such as of vitamins D and some hormones and corticosteroids. Pentacyclic hydrocarbons hopanes, as derivatives of bacteriohopantetraol which is the physiological equivalent of cholesterol, made 4.7+/-0.2% of NAN.

Gas Chromatography-Mass Spectrometry↗
PubMed1997

Pregnancy increases mobilization of lead from maternal skeleton.

The question of the extent of lead mobilization from the maternal skeleton during pregnancy and lactation is one of the most outstanding problems of lead toxicity. We have undertaken a longitudinal cohort study in an urban environment of European female immigrants of child-bearing age (18 to 35 years) to Australia whose skeletal lead isotopic composition has been determined to be different from that in their current environment. The cohort was to consist of 100 immigrants anticipated to provide 20 pregnant subjects who would be compared with two groups of control subjects: a matched immigrant nonpregnant control group and second-generation Australian pregnant control subjects. Pregnant subjects also serve as their own controls for a comparison of changes during gestation with those before conception. High-precision lead isotopic compositions and lead concentrations are measured in maternal blood and urine prenatally, monthly during gestation, and postnatally for 6 months; they are also measured in infant blood and urine for 6 months; environmental measures are sampled quarterly for 6-day duplicate diet, house dust and water, and urban air and gasoline. Because of continuing public health concerns about lead exposure, interim findings from this cohort are being reported. To date there have been 13 conceptions in immigrant subjects, with 7 births, in addition to 3 conceptions in the Australian control group, with 2 births. PbBs have been generally low, with a geometric mean of 3.0 microg/dl, and have ranged from 1.9 to 20 microg/dl. Increases in PbB of approximately 20% during pregnancy have been detectable even in subjects with low blood lead levels. The skeletal contribution to blood lead level, based on isotopic measurements, has exhibited a mean increase (and standard deviation) of 31% +/- 19% with a range from 9% to 65%. Earlier studies that used lead concentrations only have suggested that blood lead levels increased only during the second half of pregnancy. This increase in blood lead levels has also been observed in the present study. However, in two subjects the increases in total blood lead were also detected in the first 2 months of pregnancy. Changes in isotopic composition and blood lead during gestation for Australian pregnant controls were negligible. The ratio of cord/maternal blood lead levels varied from 0.54 to 1.05, and the ratio for the isotopic composition was 0.993 to 1.002. Results of this study confirm that lead is mobilized from skeletal stores at an accelerated rate during pregnancy and is transferred to the fetus. These results also show that mobilization from long-term stores (i.e., bone) contributes significantly to blood lead levels during pregnancy. Furthermore, exposure of the fetus to lead during pregnancy has implications for interpretations of neurobehavioral disorders attributed to only postnatal exposure. Even after 800 days of residence in Australia, the contribution of European skeletal lead to blood lead in nonpregnant subjects can be on the order of 50%, but the current PbB may give no indication of the former high skeletal lead burden.

Adolescent↗
PubMed1992

Gaps in the erythrocyte membrane skeleton: a stretched net model.

The geometry of spectrin-free regions in the erythrocyte membrane skeleton is modeled using Monte Carlo calculations for an incomplete triangular lattice of entropy springs under tension. Intact springs correspond to normal spectrin molecules, and cut springs correspond to spectrin that is missing or unable to associate normally. As springs are cut and the network is allowed to relax to mechanical equilibrium, gaps in the network appear. Geometrical properties of these gaps are obtained as a function of the fraction of springs cut. The most important property modeled is the area of the largest spectrin-free region; this area increases approximately exponentially as the fraction of normal spectrin decreases from 100% to approximately 50%. The effect of these gaps on lateral diffusion and vesiculation is discussed.

Erythrocyte Membrane↗
PubMed2002

Analysis of chlorinated paraffins in cutting fluids and sealing materials by carbon skeleton reaction gas chromatography.

Chlorinated paraffins (CPs) pose a major risk in the environment, due to their wide application, to their persistence, to their carcinogenic potential, and in view of the fact, that they cannot be easily identified. Various commercial cutting fluids and sealing materials were analysed for CPs with carbon skeleton reaction gas chromatography (GC) and flame ionization detection. CPs are simultaneously dechlorinated and hydrogenated to the corresponding alkanes with Pd catalyst material in the GC injector. With this method, the carbon chain length of commercial technical CPs was determined. In six of sixteen sealing materials we found five short and one medium chain length polychlorinated paraffins in percentages of 9-16% (w/w). In five cutting fluids we found predominantly medium to long chain length chloroparaffins with percentages of 2.5-31% (w/w), only one fluid contained short chain CPs.

Catalysis↗
PubMed2003

Bone autografting of the calvaria and craniofacial skeleton: historical background, surgical results in a series of 15 patients, and review of the literature.

BACKGROUND: Although the use of autologous bone for reconstruction of the cranial and facial skeleton underwent a partial reappraisal following the introduction of a vast range of alloplastic materials for this purpose, it has demonstrated definite advantages over the last century and, particularly, during the last decade. METHODS: Fifteen patients underwent cranial and/or cranio-facial reconstruction using autologous bone grafting in the Department of Neurologic Sciences-Neurosurgery and the Division of Maxillo-Facial Surgery of the Rome "La Sapienza" University between 1987 and 1995. This group of patients consisted of 8 females and 7 males whose average age was 29.5 years (range 7.5 to 59 years, mean age 30). In all these patients cranioplasty and/or cranio-facial reconstruction had been performed to repair bone defects secondary to benign tumors or tumor-like lesions (12 cases), trauma (2 cases), or, in the remaining case, to wound infection after craniotomy for a neurosurgical operation. RESULTS: The results obtained in a series of 15 patients treated using this method are described with reference to the abundant data published on this topic. CONCLUSION: The mechanical, immunologic, and technical-grafting properties of autologous bone, together with its superior esthetic and psychological effects, probably make it the best material for cranioplasty.

Adolescent↗
PubMed2001

Wnt-14 plays a pivotal role in inducing synovial joint formation in the developing appendicular skeleton.

The long bones of the vertebrate appendicular skeleton arise from initially continuous condensations of mesenchymal cells that subsequently segment and cavitate to form discrete elements separated by synovial joints. Little is known, however, about the molecular mechanisms of joint formation. We present evidence that Wnt-14 plays a central role in initiating synovial joint formation in the chick limb. Wnt-14 is expressed in joint-forming regions prior to the segmentation of the cartilage elements, and local misexpression of Wnt-14 induces morphological and molecular changes characteristic of the first steps of joint formation. Induction of an ectopic joint-like region by Wnt-14 suppresses the formation of the immediately adjacent endogenous joint, potentially providing insight into the spacing of joints.

Animals↗
PubMed1985

A monoclonal antibody inhibits calcium accumulation and skeleton formation in cultured embryonic cells of the sea urchin.

The assembly of the spicules (primitive skeleton) of the sea urchin embryo is being studied in primary mesenchyme cells cultured in vitro. A monoclonal antibody (1223) has been prepared that inhibits the deposition of CaCO3 into the spicules. This antibody reacts with a 130,000 Mr cell-surface protein that is concentrated on the surface of approximately 5% of the cells of dissociated gastrula stage embryos. When primary mesenchyme cells in the embryo or cells cultured in vitro are examined, the 1223 antigen is detected on the surface of the cells and on the extracellular material associated with the spicule. We conclude that the 1223 antibody recognizes a cell-surface protein that plays an essential role in spicule formation.

Animals↗
PubMed1976

Anabolic effect of low doses of a fragment of human parathyroid hormone on the skeleton in postmenopausal osteoporosis.

Parathyroid hormone, injected daily in low dosage, exerted anabolic effects on the human skeleton, just as it does in the rat. Four postmenopausal women with primary osteoporosis were treated for six months with a synthetic fragment of human parathyroid hormone (hP.T.H. 1-34), given as a daily injection of 100 mug. This treatment caused a remarkable acceleration of bone turnover, indicated both by isotopic tracer and histological methods. At this normocalcaemic dose level, the increases in bone formation outweighed increases in resorption. Three of the four patients showed more positive calcium balances, and mean increases in calcium and phosphorus balances were statistically significant for the group as a whole, the changes being principally due to increased intestinal absorption of both elements. Many modifications of the present method of hormone administration are possible which could further increase the preponderance of anabolic effects. These results suggest that low doses of hP.T.H. 1-34, alone or in combination with other agents, may prove useful in the treatment of osteoporosis.

Absorption↗
PubMed1979

A physiological role for calcitonin: protection of the maternal skeleton.

Plasma-calcitonin levels, measured with an established and reliable extraction radioimmunoassay technique, were significantly higher throughout normal pregnancy and lactation than in normal non-pregnant women, and were not immediately influenced by the acute stimulation of breast-feeding. Thus, more calcitonin circulates at times of physiologically increased calcium need. It is suggested that an important function of calcitonin is the protection of the healthy maternal skeleton from excessive resorption by opposing the resorptive action of 1,25-dihydroxycholecalciferol on bone.

Adolescent↗
PubMed1992

Complex orbital fracture repair using rigid fixation of the internal orbital skeleton.

Large orbital fractures involving more than one orbital wall are frequently associated with enophthalmos and vertical globe dystopia. The authors repaired 22 orbits in 20 patients using the technique of rigid fixation of the internal orbital skeleton. Eleven orbits were reconstructed with titanium or vitallium implants that were custom-shaped to span the bony defect and fixed to the orbital rim. Seven orbits were reconstructed with bone grafts rigidly supported by a miniplate, and, in four cases, direct lag screw support of bone grafts was used. The intraoperative goal was to restore the native orbital anatomy and volume. Autologous calvarial bone grafts were used to supplement the repair to achieve the desired volumetric effect and to cover additional wall defects. A reoperation for additional orbital augmentation was needed in one enophthalmic and one anophthalmic orbit. There were no cases of overcorrection, migration, infection, or extraocular muscle entrapment. In complex orbital fractures, the technique of rigid internal orbital fixation appears to yield a stable and predictable result with the prevention of postoperative globe malposition.

Adult↗