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PubMed2004

Extended neck regions stabilize tetramers of the receptors DC-SIGN and DC-SIGNR.

The human cell surface receptors DC-SIGN (dendritic cell-specific intercellular adhesion molecule-grabbing nonintegrin) and DC-SIGNR (DC-SIGN-related) bind to oligosaccharide ligands found on human tissues as well as on pathogens including viruses, bacteria, and parasites. The extracellular portion of each receptor contains a membrane-distal carbohydrate-recognition domain (CRD) and forms tetramers stabilized by an extended neck region consisting of 23 amino acid repeats. Cross-linking analysis of full-length receptors expressed in fibroblasts confirms the tetrameric state of the intact receptors. Hydrodynamic studies on truncated receptors demonstrate that the portion of the neck of each protein adjacent to the CRD is sufficient to mediate the formation of dimers, whereas regions near the N terminus are needed to stabilize the tetramers. Some of the intervening repeats are missing from polymorphic forms of DC-SIGNR. Two different crystal forms of truncated DC-SIGNR comprising two neck repeats and the CRD reveal that the CRDs are flexibly linked to the neck, which contains alpha-helical segments interspersed with non-helical regions. Differential scanning calorimetry measurements indicate that the neck and CRDs are independently folded domains. Based on the crystal structures and hydrodynamic data, models for the full extracellular domains of the receptors have been generated. The observed flexibility of the CRDs in the tetramer, combined with previous data on the specificity of these receptors, suggests an important role for oligomerization in the recognition of endogenous glycans, in particular those present on the surfaces of enveloped viruses recognized by these proteins.

Animals↗
PubMed2005

Increased flexibility as a strategy for cold adaptation: a comparative molecular dynamics study of cold- and warm-active uracil DNA glycosylase.

Uracil DNA glycosylase (UDG) is a DNA repair enzyme in the base excision repair pathway and removes uracil from the DNA strand. Atlantic cod UDG (cUDG), which is a cold-adapted enzyme, has been found to be up to 10 times more catalytically active in the temperature range 15-37 degrees C as compared with the warm-active human counterpart. The increased catalytic activity of cold-adapted enzymes as compared with their mesophilic homologues are partly believed to be caused by an increase in the structural flexibility. However, no direct experimental evidence supports the proposal of increased flexibility of cold-adapted enzymes. We have used molecular dynamics simulations to gain insight into the structural flexibility of UDG. The results from these simulations show that an important loop involved in DNA recognition (the Leu(272) loop) is the most flexible part of the cUDG structure and that the human counterpart has much lower flexibility in the Leu(272) loop. The flexibility in this loop correlates well with the experimental k(cat)/K(m) values. Thus, the data presented here add strong support to the idea that flexibility plays a central role in adaptation to cold environments.

Adaptation, Physiological↗
PubMed2005

An active enzyme constructed from a 9-amino acid alphabet.

Nature employs a set of 20 amino acids to produce a repertoire of protein structures endowed with sophisticated functions. Here, we combined design and selection to create an enzyme composed entirely from a set of only 9 amino acids that can rescue auxotrophic cells lacking chorismate mutase. The simplified protein captures key structural features of its natural counterpart but appears to be somewhat less stable and more flexible. The potential of a dramatically reduced amino acid alphabet to produce an active catalyst supports the notion that primordial enzymes may have possessed low amino acid diversity and suggests that combinatorial engineering strategies, such as the one used here, may be generally applied to create enzymes with novel structures and functions.

Amino Acid Sequence↗
PubMed2006

Crystallographic evidence that the dinuclear copper center of tyrosinase is flexible during catalysis.

At high resolution, we determined the crystal structures of copper-bound and metal-free tyrosinase in a complex with ORF378 designated as a "caddie" protein because it assists with transportation of two CuII ions into the tyrosinase catalytic center. These structures suggest that the caddie protein covers the hydrophobic molecular surface of tyrosinase and interferes with the binding of a substrate tyrosine to the catalytic site of tyrosinase. The caddie protein, which consists of one six-strandedbeta-sheet and one alpha-helix, has no similarity with all proteins deposited into the Protein Data Bank. Although tyrosinase and catechol oxidase are classified into the type 3 copper protein family, the latter enzyme lacks monooxygenase activity. The difference in catalytic activity is based on the structural observations that a large vacant space is present just above the active center of tyrosinase and that one of the six His ligands for the two copper ions is highly flexible. These structural characteristics of tyrosinase suggest that, in the reaction that catalyzes the ortho-hydroxylation of monophenol, one of the two Cu(II) ions is coordinated by the peroxide-originated oxygen bound to the substrate. Our crystallographic study shows evidence that the tyrosinase active center formed by dinuclear coppers is flexible during catalysis.

Amino Acid Sequence↗
PubMed2006

Effect of DNA modifications on DNA processing by HIV-1 integrase and inhibitor binding: role of DNA backbone flexibility and an open catalytic site.

Integration of the viral cDNA into host chromosomes is required for viral replication. Human immunodeficiency virus integrase catalyzes two sequential reactions, 3'-processing (3'-P) and strand transfer (ST). The first integrase inhibitors are undergoing clinical trial, but interactions of inhibitors with integrase and DNA are not well understood in the absence of a co-crystal structure. To increase our understanding of integrase interactions with DNA, we examined integrase catalysis with oligonucleotides containing DNA backbone, base, and groove modifications placed at unique positions surrounding the 3'-processing site. 3'-Processing was blocked with substrates containing constrained sugars and alpha-anomeric residues, suggesting that integrase requires flexibility of the phosphodiester backbone at the 3'-P site. Of several benzo[a]pyrene 7,8-diol 9,10-epoxide (BaP DE) adducts tested, only the adduct in the minor groove at the 3'-P site inhibited 3'-P, suggesting the importance of the minor groove contacts for 3'-P. ST occurred in the presence of bulky BaP DE DNA adducts attached to the end of the viral DNA suggesting opening of the active site for ST. Position-specific effects of these BaP DE DNA adducts were found for inhibition of integrase by diketo acids. Together, these results demonstrate the importance of DNA structure and specific contacts with the viral DNA processing site for inhibition by integrase inhibitors.

Base Sequence↗
PubMed2006

Energetics of ligand-induced conformational flexibility in the lactose permease of Escherichia coli.

Isothermal titration calorimetry has been applied to characterize the thermodynamics of ligand binding to wild-type lactose permease (LacY) and a mutant (C154G) that strongly favors an inward facing conformation. The affinity of wild-type or mutant LacY for ligand and the change in free energy (DeltaG) upon binding are similar. However, with the wild type, the change in free energy upon binding is due primarily to an increase in the entropic free energy component (TDeltaS), whereas in marked contrast, an increase in enthalpy (DeltaH) is responsible for DeltaG in the mutant. Thus, wild-type LacY behaves as if there are multiple ligand-bound conformational states, whereas the mutant is severely restricted. The findings also indicate that the structure of the mutant represents a conformational intermediate in the overall transport cycle.

Calorimetry↗
PubMed2002

Changes in expansion and mechanical strength during water storage of a traditional and three modified resin composites.

Four types of resin composites (Dyract AP (DYR), Definite (DEF), Ariston pHc (ARI), and Spectrum TPH (SPE)) were tested after water storage for up to 180 days. The test parameters were flexural strength, flexural modulus, and marginal gaps at fillings in cylindrical cavities. The cavity tests were performed in two ways. One series of filings was finished shortly after curing and the maximal gaps were followed with time during water storage. Another series was finished immediately before gap size measurement and after water storage for various periods. Absence of or reduced gap sizes at 180 days compared to 1 h or 1 day was obsenred for all materials and with both methods. At 180 days, DYR and ARI showed significantly smaller gap sizes than those observed with DEF and SPE. No significant differences in flexural strength were observed when means at 180 days were compared to those observed at 1 day. DEF, ARI, and SPE showed a significant increase in flexural moduli at 180 days compared to 1 day, while DYR showed none. It is concluded that fillings made of Dyract AP or Ariston pHc may show absence of or very small gap sizes between filling and cavity wall after 6 months and with no significant reduction in mechanical strength.

Analysis of Variance↗
PubMed2003

Flexural strength and modulus of a novel ceramic restorative cement intended for posterior restorations as determined by a three-point bending test.

The aim of this study was to compare a new restorative cement intended for posterior restorations, Doxadent, with other types of tooth-colored materials as regards flexural strength and flexural modulus. The new restorative material consists mainly of calcium aluminate. Four hybrid resin composites, one polyacid-modified resin composite, one resin-modified glass ionomer cement, one conventional glass ionomer cement, one zinc phosphate cement, and an experimental version as well as the marketed version of Doxadent were investigated. Flexural strength and flexural modulus were tested according to ISO standard 4049 and determined after 1 d, 1 week, and 2 weeks. Together with the zinc phosphate cement, Doxadent had the lowest flexural strengths (13-22 MPa). The strongest materials were the resin composites and the polyacid-modified resin composite (83-136 MPa). The highest flexural modulus was found for Doxadent (17-19 GPa). The flexural strength of Doxadent decreased significantly from 1 week to 2 weeks, while flexural modulus remained unchanged. The other materials reacted in different ways to prolonged water storage. It can be concluded that the restorative cement Doxadent had significantly lower flexural strength and significantly higher flexural modulus than today's materials used for direct posterior restorations.

Aluminum Compounds↗
PubMed2004

Flexural strength and modulus of a step-cured resin composite.

In an attempt to reduce marginal contraction gaps, a step-curing mode of light polymerization of resin composite has been proposed. It was hypothesized that such an exposure mode, having an initial reduced curing rate, would result in composite having a lower modulus of elasticity than composite exposed in one step. The composite was initially exposed to power density levels of 50, 100, 150, or 200 mW/cm2 for durations of 10, 20, or 40 s. The final exposure and the exposure of the control group were performed at 750 mW/cm2 for 20 s. It was found that certain modes of two-step exposure resulted in a lower flexural modulus than did a one-step exposure of constant, high power density. Regression analysis showed, with statistical significance, that flexural modulus was relatively high following a short initial exposure at low power density or using a long initial exposure at high power density. Conversely, flexural modulus was relatively low following a long initial exposure at low power density or using a short initial exposure at high power density. The results were explained by probable differences in degree of conversion and cross-link density of the polymer network.

Analysis of Variance↗
PubMed2006

Effect of surface preparation using ethyl acetate on the repair strength of denture base resin.

OBJECTIVE: The objective of this study was to evaluate the effect of surface preparation using ethyl acetate on the repair strength of a denture base resin. MATERIAL AND METHODS: Bar specimens were fabricated with a heat-processed denture base resin and cut in half. The repair surfaces of the specimens were treated with ethyl acetate for varying amounts of time before the autopolymerizing repair resin was applied. The specimens were then immersed in 37 degrees C distilled water for 2 days. A three-point bending test was used to determine the transverse strength of the test specimens. Ten specimens prepared using a 120-s application of ethyl acetate and 10 specimens prepared using a 5-s application of dichloromethane were thermocycled at 10,000 cycles (at 5 degrees C and 55 degrees C) with a 60-s dwell time. The morphological changes in the repair surfaces after preparation were observed by scanning electron microscopy. RESULTS: The repair strength of the denture base resin prepared using ethyl acetate for 120 s (30.6 MPa) was the highest among the results for the various amounts of time tested (p<0.05). There were no significant differences between the repair bond strength both before and after thermocycling of the denture base resin prepared using ethyl acetate for 120 s and that of the denture base resin prepared with dichloromethane for 5 s (p>0.05). CONCLUSION: Within the limitations of the current study, a 120-s application of ethyl acetate was as effective as a 5-s application of dichloromethane at preparing the surfaces of a denture base resin.

Acetates↗
PubMed2005

Effect of deep-freezing on some properties of acrylic resin.

The objective of this study was to evaluate the effect of freezing the acrylic resin polymer/monomer dough mix on a range of properties of the polymerized denture base material. Powder and liquid of acrylic resin were mixed and at the dough stage transferred to a deep freezer and frozen for periods of 24 h, 1 week, 1 month, 3 months, and 6 months. At the end of the storage time, sufficient dough was removed, thawed, packed, and polymerized, and specimens were prepared from the polymerized plates. A control series was prepared from dough which had followed conventional mixing and packing procedures without freezing. The strength properties, hardness, and flash thickness were examined. Statistical analysis was carried out using ANOVA (one-way and two-way) with multiple range tests. The flexural strength was significantly (p=0.03) increased and the impact strength, hardness, and flash thickness were not affected by freezing the material during the dough stage. The flexural modulus was significantly (p=0.0001) reduced when the storage time in the freezer was increased to 3 months. Six months' frozen material proved to be unpackable. It was concluded that the acrylic resin polymer/monomer dough mix can be stored in the freezer for up to 1 month without any statistically significant effect on the properties of the polymerized denture base material.

Acrylic Resins↗
PubMed2005

Evaluation of some properties of two fiber-reinforced composite materials.

OBJECTIVE: Water sorption, flexural properties, bonding properties, and elemental composition of photopolymerizable resin-impregnated fiber-reinforced composite (FRC) materials (everStick C&B and BR-100) (FPD) were evaluated in this study. MATERIAL AND METHODS: Bar-shaped specimens (2 x 2 x 25 mm) were prepared for water sorption and flexural strength testing. The specimens (n = 6) were polymerized either with a hand light-curing unit for 40 s or, additionally, in a light-curing oven for 20 min and stored in water for 30 days. Water sorption was measured during this time, followed by measurements of flexural strength and modulus. A shear bond strength test was performed to determine the bonding characteristics of polymerized FRC to composite resin luting cement (Panavia-F), (n = 15). The cement was bonded to the FRC substrate and the specimens were thermocycled 5000 times (5-55 degrees C) in water. SEM/EDS were analyzed to evaluate the elemental composition of the glass fibers and the fiber distribution in cross section. RESULTS: ANOVA showed significant differences in water sorption according to brand (p < 0.05). Water sorption of everStick C&B was 1.86 wt% (hand-unit polymerized) and 1.94 wt% (oven polymerized), whereas BR-100 was 1.07 wt% and 1.17 wt%, respectively. The flexural strength of everStick C&B after 30 days' water storage was 559 MPa (hand-unit polymerized) and 796 MPa (oven-polymerized); for BR-100, the values were 547 MPa and 689 MPa, respectively. Mean shear bond strength of composite resin cement to the FRC varied between 20.1 and 23.7 MPa, showing no statistical difference between the materials. SEM/EDS analysis revealed that fibers of both FRC materials consist of the same oxides (SiO2, CaO, and Al2O3) in ratios. The distribution of fibers in the cross section of specimens was more evenly distributed in everStick C&B than in BR-100. CONCLUSION: The results of this study suggest that there are some differences in the tested properties of the FRC materials.

Aluminum Oxide↗
PubMed2006

Effect of variations from the recommended powder/liquid ratio on some properties of resin-modified cements.

OBJECTIVES: The powder and liquid contents of cements are mixed in accordance with the recommended mixing ratio, but discrepancies occur despite the use of proportioning scoops. Little is known about powder/liquid ratio variations on certain properties of resin-modified cements. METHODS: Two resin-modified glass ionomer cements (RMGIC) were mixed using various powder/liquid ratios: (a) Recommended ratio: Fuji Plus: powder/liquid 2:1; ProTecCEM p/l 2.25:1. (b) Maximal variation arising using proportioning aids (17% liquid surplus): Fuji Plus 1.66:1, ProTecCEM 1.875:1. Limits of mixing and specimen construction: either (c) both groups with more liquid (1.5:1) or (d) more powder (3:1). Flexural strength was determined using a 3-point bending test (after 24 h) and wear using a 3-body abrasion device. The extent of cure reaction was characterized using differential thermal analysis (DTA). RESULTS: While higher powder content did not significantly affect the flexural strength of Fuji Plus and ProTecCEM, it considerably reduced wear of Fuji Plus. Increasing liquid content reduced flexural strength. A substantial increase in wear for Fuji Plus 1.5:1, and ProTecCEM 1.875:1 and 1.5:1 mixtures was observed. DTA demonstrated that a higher liquid content resulted in incomplete setting reactions, which could be detected even after 24 h of cure. CONCLUSIONS: If RMGICs are mixed with powder/liquid variations, given the inaccuracy of proportioning aids the properties of RMGIC will change slightly and may be disregarded. If set with higher powder/liquid variations, a surplus of powder has less influence on the properties than a surplus of liquid.

Dental Restoration Wear↗
PubMed2006

Effect of continuous longitudinal glass fiber reinforcement on the cantilever beam strength of particulate filler composites.

OBJECTIVE: The objective of this in vitro study was to investigate the effect of continuous longitudinal glass fibers on the bending properties of particulate filler composite resins. MATERIAL AND METHODS: Four particulate filler composite resins (Tetric Ceram, Point 4, Z250, P60) and one continuous longitudinal glass fiber material (everStick) as reinforcement were selected. Three groups of beam specimens (5 x 5 x 25 mm) for each material were fabricated. For group I, specimens were not reinforced with fibers and were tested as the control group. For group II, specimens were reinforced with 2 layers of the fiber (6.5 vol%), and for group III with 3 layers (9.8 vol%). The specimens were stored at 37 degrees C for 30 days. One end of the beam specimen (14 mm) was fixed in a device, while the other (11 mm) was left free. Cantilever beam strength was measured using a universal testing machine, loading at a distance of 10 mm from the upright support through a steel ball of 2 mm diameter. The loads and deflection at initial failure and at final failure for each specimen were recorded. The data were statistically analyzed using one-way ANOVA and the multiple comparison Scheffé test (alpha = 0.05). RESULTS: The bending moments of particulate filler composite at final failure, when reinforced with 3 layers of the glass fibers (272.4-325.2 Ncm), were significantly higher than for the composites without fibers. However, the materials reinforced with 3 layers of fibers were not significantly different from the materials reinforced with 2 layers of fibers (234.1-282.6 Ncm). The materials reinforced with 3 layers of fibers exhibited severe deflection at final failure, ranging from 2.8 mm to 3.4 mm. The bending moments of the particulate composites increased linearly with the weight fraction of the fillers, but there was no linear correlation between them when reinforced with fibers. CONCLUSIONS: The cantilever beam strength of the particulate filler composites increased significantly when layers of fibers were added and as the weight fraction of filler increased, but a higher fiber volume fraction did not lead to a significantly higher cantilever beam strength.

Composite Resins↗
PubMed2004

Platelet concentrate injection improves Achilles tendon repair in rats.

BACKGROUND: Blood platelets release a cocktail of growth factors when activated, some of which are thought to initiate and stimulate repair. EXPERIMENT AND FINDINGS: We studied whether a platelet concentrate injection would improve Achilles tendon repair in an established rat model. The Achilles tendon was transected and a 3 mm segment removed. After 6 h, a platelet concentrate was injected percutaneously into the hematoma. This increased tendon callus strength and stiffness by about 30% after 1 week, which persisted for as long as 3 weeks after the injection. At this time, the mechanical testing indicated an improvement in material characteristics--i.e., greater maturation of the tendon callus. This was confirmed by blinded histological scoring. INTERPRETATION: Platelet concentrate may prove useful for the treatment of Achilles tendon ruptures.

Achilles Tendon↗
PubMedDate not supplied

Comparison of muscle strength and flexibility between the preferred and non-preferred leg in English soccer players.

Most soccer players have a favoured foot for kicking the ball, and it is believed that this preference may lead to an asymmetry in the strength and flexibility of the lower extremities. This study was designed to determine whether asymmetry in strength and flexibility are present in the legs of soccer players.Forty-one elite and sub-elite soccer players (age 23.4 +/- 3.8 years; height 1.81 +/- 0.06 m; body mass 81.7 +/- 9.9 kg) were studied (data are presented as mean +/- SD). The dynamic strength of knee flexors (hamstrings) and knee extensors (quadriceps) was measured using an isokinetic dynamometer at angular velocities of 1.05, 2.09, 5.23 rad/s (in a concentric mode) and 2.09 rad/s (in an eccentric mode). The concentric strength ratio (hamstrings(conc)/quadriceps(conc)) and the dynamic control ratio (hamstrings(ecc)/quadriceps(conc)) were computed. Hip joint flexibility (in flexion) was measured using a goniometer.A significant difference between the preferred and non-preferred leg was found in the knee flexors at 2.09 rad/s (119 +/- 22 versus 126 +/- 24 Nm; P < 0.05) and for the dynamic control ratio (0.79 +/- 0.13 versus 0.84 +/- 0.16 Nm; P < 0.05). In both cases the knee flexors of the preferred leg were weaker than those of the non-preferred leg. A total of 28 of the 41 players (68%) had significant musculoskeletal abnormality (imbalance >10%) in one or more specific muscle groups. No significant differences were found in flexibility of the hip joint between the preferred and non-preferred leg (P > 0.05). It is concluded that the lower strength of the knee flexor muscles of the preferred leg may be associated with the differential use of these muscle during the kicking action and thus constitutes a unique training effect associated with soccer. This in turn can lead to muscular imbalance which is generally regarded as an injury risk factor.

Adult↗
PubMed2001

A comparison of the anthropometric, strength, endurance and flexibility characteristics of female elite and recreational climbers and non-climbers.

There is limited information on the anthropometry, strength, endurance and flexibility of female rock climbers. The aim of this study was to compare these characteristics in three groups of females: Group 1 comprised 10 elite climbers aged 31.3 +/- 5.0 years (mean +/- s) who had led to a standard of 'hard very severe'; Group 2 consisted of 10 recreational climbers aged 24.1 +/- 4.0 years who had led to a standard of 'severe'; and Group 3 comprised 10 physically active individuals aged 28.5 +/- 5.0 years who had not previously rock-climbed. The tests included finger strength (grip strength, finger strength measured on climbing-specific apparatus), flexibility, bent arm hang and pull-ups. Regression procedures (analysis of covariance) were used to examine the influence of body mass, leg length, height and age. For finger strength, the elite climbers recorded significantly higher values (P < 0.05) than the recreational climbers and non-climbers (four fingers, right hand: elite 321 +/- 18 N, recreational 251 +/- 14 N, non-climbers 256 +/- 15 N; four fingers, left hand: elite 307 +/- 14 N, recreational 248 +/- 12 N, non-climbers 243 +/- 11 N). For grip strength of the right hand, the elite climbers recorded significantly higher values than the recreational climbers only (elite 338 +/- 12 N, recreational 289 +/- 10 N, non-climbers 307 +/- 11 N). The results suggest that elite climbers have greater finger strength than recreational climbers and non-climbers.

Abdominal Muscles↗
PubMed2002

Comparison of spine motion in elite golfers with and without low back pain.

Low back pain is a common musculoskeletal disorder affecting golfers, yet little is known of the specific mechanisms responsible for this injury. The aim of this study was to compare golf swing spinal motion in three movement planes between six male professional golfers with low back pain (age 29.2+/-6.4 years; height 1.79+/-0.04 m; body mass 78.2+/-12.2 kg; mean +/- s) and six without low back pain (age 32.7+/-4.8 years; height 1.75+/-0.03 m; body mass 85.8+/-10.9 kg) using a lightweight triaxial electrogoniometer. We found that golfers with low back pain tended to flex their spines more when addressing the ball and used significantly greater left side bending on the backswing. Golfers with low back pain also had less trunk rotation(obtained from a neutral posture), which resulted in a relative 'supramaximal' rotation of their spines when swinging. Pain-free golfers demonstrated over twice as much trunk flexion velocity on the downswing, which could relate to increased abdominal muscle activity in this group. This study is the first to show distinct differences in the swing mechanics between golfers with and without low back pain and provides valuable guidance for clinicians and teachers to improve technique to facilitate recovery from golf-related low back pain.

Adult↗