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Background and Purpose: Ca(2+)-dependent Cl(-) secretion (CaCC) in airways and other tissues is due to activation of the Cl(-) channel TMEM16A (anoctamin 1). Earlier studies suggested that Ca(2+) -activated Cl(-) channels are regulated by membrane lipid inositol phosphates, and that 1-O-octyl-2-O-butyryl-myo-inositol 3,4,5,6-tetrakisphosphate octakis(propionoxymethyl) ester (INO-4995) augments CaCC. Here we examined whether TMEM16A is the target for INO-4995 and if the channel is regulated by inositol phosphates.Experimental Approach: The effects of INO-4995 on CaCC were examined in overexpressing HEK293, colonic and primary airway epithelial cells as well as Xenopus oocytes. We used patch clamping, double electrode voltage clamp and Ussing chamber techniques.Key Results: We found that INO-4995 directly activates a TMEM16A whole cell conductance of 6.1 ± 0.9 nS pF(-1) in overexpressing cells. The tetrakisphosphates Ins(3,4,5,6)P(4) or Ins(1,3,4,5)P(4) and enzymes controlling levels of InsP(4) or PIP(2) and PIP(3) had no effects on the magnitude or kinetics of TMEM16A currents. In contrast in Xenopus oocytes, human airways and colonic cells, which all express TMEM16A endogenously, Cl(-) currents were not acutely activated by INO-4995. However incubation with INO-4995 augmented 1.6- to 4-fold TMEM16A-dependent Cl(-) currents activated by ionomycin or ATP, while intracellular Ca(2+) signals were not affected. The potentiating effect of INO-4995 on transient ATP-activated TMEM16A-currents in cystic fibrosis (CF) airways was twice of that observed in non-CF airways.Conclusions and Implications: These data indicate that TMEM16A is the target for INO-4995, although the mode of action appears different for overexpressed and endogenous channels. INO-4995 may be useful for the treatment of CF lung disease. [ABSTRACT FROM AUTHOR]

We numerically investigated the use of graphene nanoribbons placed on top of silicon-on-insulator (SOI) strip waveguides for light polarization control in silicon photonic-integrated waveguides. We found that two factors mainly affected the polarization control: the graphene chemical potential and the geometrical parameters of the waveguide, such as the waveguide and nanoribbon widths and distance. We show that the graphene chemical potential influences both TE and TM polarizations almost in the same way, while the waveguide width tapering enables both TE-pass and TM-pass polarizing functionalities. Overall, by increasing the oxide spacer thickness between the silicon waveguide and the top graphene layer, the device insertion losses can be reduced, while preserving a high polarization extinction ratio.

In this work, we show the design of a silicon photonic-based polarization converting device based on the integration of semiconduction InP nanowires on the silicon photonic platform. We present a comprehensive numerical analysis showing that full polarization conversion (from quasi-TE modes to quasi-TM modes, and vice versa) can be achieved in devices exhibiting small footprints (total device lengths below 20 µm) with minimal power loss (<2 dB). The approach described in this work can pave the way to the realization of complex and re-configurable photonic processors based on the manipulation of the state of polarization of guided light beams.

The aim of this study was to determine Vickers microhardness (HV) in bulk fill resins at different depths. Test specimens were prepared with different bulk fill resins: Filtek Bulk-Fill (3M ESPE) [FBF], Surefill SDR flow (Dentsply) [SDR], Fill-UP (COLTENE) [FU] and Surefill (Dentsply) [SF]. Semi-cylindrical test specimens were prepared in a mold 6 mm in diameter and 4 mm thick (n=5). A 1000 mW/cm2 light curing unit was applied (Coltolux LED - Coltene) for 20 seconds. HV was determined with three indentations (Vickers Future Tech FM300, 300 g, 8 s) at four depths: 1, 2, 3 and 4 mm from the top surface to the interior. Data were recorded immediately (t0) and 24 hours later (t24). Results were analyzed with two-way ANOVA (p<0.05), and multiple comparisons were performed using Tukey's test. Mean and SD of HV at t0 for each mm were: [FBF] t0: 49.23(4.65) / 48.32(3.36) / 44.38(2.06) / 40.59(2.58); [FBF] t24: 61.37(3.47) / 62.63(3.03) / 57.27(5.22) / 56.37(5.88);[SDR]t0:27.81(3.13) / 28.07(2.4) / 27.24(2.94) / 25.71(3.0); [SDR] t24: 35.11(2.16) / 35.17(1.96) / 35.53(1.81) / 33.18(2.08); [FU] t0: 41.43(1.41) / 39.87(0.88) / 38.11(1.81) / 39.09(1.92); [FU] t24: 49.27(1.54) / 48.77(1.77) / 48.65(1.88) / 46.76(4.93); [SF] t0: 71.35(7.09) / 67.39(9.76) / 68.95(6.21) / 64.1(8.35); [SF] t24: 76.06(6.61) / 75.31(9.37) / 75.2(11.57) / 69.81(12.14). ANOVA showed significant effect of material, depth and recording time (p<0.05), and Tukey's test showed that recording sites (depths) differed significantly, giving four homogeneous groups. Under the conditions of this study, it can be concluded that microhardness of bulk-fill resins can be affected by depth and post-curing time., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Sociedad Argentina de Investigación Odontológica.)

The aim of this research was to determine compressive and shear bond strength of blocks prepared with bulk-fill and nanofill composite resin combinations. Materials used were Filtek Bulk Fill (FBF) and Z350 (both 3M-ESPE) and Surefil SDR flow (SFF) - Dentsply. To determine shear bond strength, cylindrical specimens 10 mm thick were prepared with composite consisting of thicknesses of 6 mm of one material and 4 mm of the other, in the following combinations: G1: FBF- FBF; G2: Z350-Z350, G3: FBF-Z350, G4: Z350-SFF and G5: SFFSFF. Materials were cured using a 1100 mw/cm2 light for 20 seconds for each layer. Samples were stored for 24 hours at 37 °C in distilled water and shear bond strength was determined. To assess compressive strength, cylindrical samples 4 mm diameter and 6 mm thick consisting of 4 mm + 2 mm were used in the same combinations as described above, stored in distilled water at 37 °C for 24 hours, after which compressive strength was determined. Both tests were performed with a Universal testing machine at a cross head speed of 1 mm/min. Results were analyzed with ANOVA and Tukey's test. Means and standard deviations in MPa for each group were the following: Shear bond strength: G1: 435.87 (65.86), G2: 233.6 (108.15), G3: 279.2 (22.05), G4:449.1 (109.35) and G5: 196.6 (51.16). Compressive strength: G1:160.07(4.27), G2: 149.49 (14.06), G3: 156.10 (29.99), G4: 199-30(39.28), G5: 171.23 (28.71). Evaluation with ANOVA showed no significant differences among combinations for compressive strength (p>0.05) and significant differences for bond strength (p<0.05). Tukey's test showed three homogeneous groups. Under these experimental conditions, it can be concluded that the study combinations have adequate mechanical behavior, equivalent to materials used individually. However, shear bond strength was affected by the combinations analyzed., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article, (Sociedad Argentina de Investigación Odontológica.)

The aim of this study was to evaluate the effect of flowable composite or glass ionomer liners on the shrinkage stress of a restorative composite resin. Fifteen previously sandblasted metal boxes were attached to a universal mechanical testing machine (INSTRON 1011, Instron Corporation). Five of these boxes were filled with Filtek Z350 XT (FXT) Universal Restorative A2 (3M ESPE) (Group 1 or Control). Two further groups of 5 boxes were prepared by interposing a layer of Vitrebond Light Cure Glass Ionomer 3M ESPE (VGI) (Group 2 or G.I.) or Filtek Z350 XT Flowable Restorative A2 3M ESPE (FFR) (Group 3 or Flowable) between the box and the composite resin, completing with the same volume of composite as in Group 1. Upon activating lightcuring, the filled boxes mounted on the testing machine were videoed for 60 seconds (40 s photoactivation and 20 s postcuring), timed with a digital chronometer. Force values were recorded in newtons and converted into stress according to contact surface. Stress values were recorded every 10 s. Results were analyzed using repeated measures ANOVA. Mean and standard deviation in kPa (stress) recorded for each group were: Control group: 126.2 (30.8); G.I.: 48.4 (18); Flowable: 27.9 (19.5). Statistical analysis showed significant differences between the control group and the rest (p<0.01), with no significant difference between groups with glass ionomer liners and flowable resin liners (G.I. and Flowable). Under the experimental conditions of this study, it can be concluded that polymerization shrinkage stress can be reduced by the presence of a liner between the preparation and the restorative material.

Optical technology applied to on-chip wireless communication is particularly promising to overcome the performance limitations of the state-of-the-art networks on-chip. A key enabling component for such applications is the plasmonic antenna coupled to conventional silicon waveguides, which can guarantee full compatibility with standard optical circuitry. In this paper, we propose an antenna array configuration based on tilted plasmonic Vivaldi antennas coupled to a silicon waveguide. The details of the single antenna and of the array design are reported. The radiation characteristics of the array are suitable for on-chip point-to-point communication, i.e. in-plane maximum gain of 14.70 dB for an array with five antennas. The array exploits a travelling wave feeding scheme and, therefore, is compact in size (about 3.5 µm × 8.7 µm ).

In this paper we propose a novel hybrid optical plasmonic Vivaldi antenna for operation in the standard C telecommunication band for wavelengths in the 1550 nm range. The antenna is fed by a silicon waveguide and is designed to have high gain and large bandwidth. The shape of the radiation pattern, with a main lobe along the antenna axis, makes this antenna suitable for point-to-point connections for inter- or intra-chip optical communications. Direct port-to-port short links for different connection distances and in a homogeneous environment have also been simulated to verify, by comparing the results of a full-wave simulation with the Friis transmission equation, the correctness of the antenna parameters obtained via near-to-far field transformation.

Endoplasmic reticulum-plasma membrane (ER-PM) contact sites play an integral role in cellular processes such as excitation-contraction coupling and store-operated calcium entry (SOCE). Another ER-PM assembly is one tethered by the extended synaptotagmins (E-Syt). We have discovered that at steady state, E-Syt2 positions the ER and Sac1, an integral ER membrane lipid phosphatase, in discrete ER-PM junctions. Here, Sac1 participates in phosphoinositide homeostasis by limiting PM phosphatidylinositol 4-phosphate (PI(4)P), the precursor of PI(4,5)P2 Activation of G protein-coupled receptors that deplete PM PI(4,5)P2disrupts E-Syt2-mediated ER-PM junctions, reducing Sac1's access to the PM and permitting PM PI(4)P and PI(4,5)P2to recover. Conversely, depletion of ER luminal calcium and subsequent activation of SOCE increases the amount of Sac1 in contact with the PM, depleting PM PI(4)P. Thus, the dynamic presence of Sac1 at ER-PM contact sites allows it to act as a cellular sensor and controller of PM phosphoinositides, thereby influencing many PM processes., (© 2016 Dickson et al.)

The aim of this study was to evaluate the linear dimensional stability of different irreversible hydrocolloid materials over time. A metal mold was designed with custom trays made of thermoplastic sheets (Sabilex, sheets 0.125 mm thick). Perforations were made in order to improve retention of the material. Five impressions were taken with each of the following: Kromopan 100 (LASCOD) [AlKr], which has dimensional stability of 100 hours, and Phase Plus (ZHERMACK) [AlPh], which has dimensional stability of 48 hours. Standardized digital photographs were taken at different time intervals (0, 15, 30, 45, 60, 120 minutes; 12, 24 and 96 hours), using an "ad-hoc" device. The images were analyzed with software (UTHSCSA Image Tool) by measuring the distance between intersection of the lines previously made at the top of the mold. The results were analyzed by ANOVA for repeated measures. Initial and final values were (mean and standard deviation): AlKr: 16.44 (0.22) and 16.34 (0.11), AlPh: 16.40 (0.06) and 16.18 (0.06). Statistical evaluation showed significant effect of material and time factors. Under the conditions in this study, time significantly affects the linear dimensional stability of irreversible hydrocolloid materials., (Sociedad Argentina de Investigación Odontológica.)

The aim of this study was to determine the influence of the finishing protocol (FP) on flexural properties of several composites (CR). Twenty composite samples (25x2x2 mm) were prepared: G1 HelimolarR; G2 Filtek™ Z350;G3TetricR N Ceram, G4 Point 4™, G5 Premisa™; G6 Esthet.XR HD, G7 ice, G8 Vit-L-escenceR, G9 GrandioR, G10 TPHR3, G11 AmelogenR Plus, G12 Brilliant Enamel; G13 Filtek™ Z100 and randomly divided into four groups according to the finishing system: C control, J JiffyR, SS Super SnapR, AA AstropolR /AstrobrushR. Each sample was polished for 10 seconds with each sequence instrument, and stored in distilled water for 24 hours, after which a three-point flexure test was applied to determine flexural strength (FS) and modulus (Flexural modulus). Data were analyzed using a two-way multivariate ANOVA and means were compared with Tukey's test. Results were: FS level CR p=0.000 with significant differences. FS level FPp= 0.093 with significant differences. In order: FM level CR p 0.00 with significant differences. FM level PS p=0.001; with significant differences. Under the study conditions, the polishing systems based on silicone rubber decreased the flexural properties of composite resins.

The aim of this study was to evaluate in vitro the marginal microleakage of two glass ionomer materials used as pit and fissure sealants. Thirty healthy premolars extracted for orthodontic treatment were randomly assigned to two groups (n=15) and respectively sealed with two glass ionomers (Group I, Fuji VII and Group II, Fuji IX). All teeth were preserved in artificial saliva (NAF) for 10 days, thermocycled (250 cycles; 5 °C, 37 °C and 60 °C), isolated, and immersed in 2% alcohol gentian violet blue solution for 24 h. After washing, teeth were included in acrylic resin and sectioned longitudinally in a bucco-lingual direction with a Struers-Minitom cutting device. Samples were analyzed for leakage using an optical microscope (Olympus BX- 60M). TheWilliams and Winter semi-quantitative ranked scale was used to score dye penetration. In Group I the grades were distributed as follows: Grade 1, 1 sample and Grade 3, 14 samples (Mean 2.87 Median 3, SD 0.52). In Group II: Grade 0: 4 samples, Grade 1, 3 samples, Grade 2, 2 samples and Grade 3, 6 samples (Mean 1.67, Median 2, SD 1.29). Fisher's exact test showed statistically significant differences between materials (p=0.006). From these results, we conclude that Fuji IX had better marginal sealing than Fuji VII when used as a pit and fissure sealant.

An overdense plasma layer irradiated by intense light can exhibit dramatic nonlinear-optical effects due to a relativistic mass-effect of free electrons: highly-multiple hysteresises of reflection and transition, and emergence of immobile waves of large amplitude. Those are trapped quasi-soliton spikes sustained by a weak pumping having a tiny fraction of their peak intensity once they have been excited first by higher power pumping. The phenomenon persists even in the layers with "soft", wash-out boundaries, as well as in a semi-infinite plasma with low absorption.

Wave transport in a media with a slow spatial gradient of its characteristics is found to exhibit a universal wave pattern ("gradient marker") in a vicinity of the maxima or minima of the gradient. The pattern is common for optics, quantum mechanics, and any other propagation governed by the same wave equation. Derived analytically, it has an elegantly simple yet nontrivial profile found in perfect agreement with numerical simulations for specific examples. We also find resonant states in continuum in the case of quantum wells, and formulated criteria for their existence.

Unlabelled: The purpose of this study was to evaluate the linear dimensional stability of different elastomeric impression materials over time. A metal mold was designed with its custom trays, which were made of thermoplastic sheets (Sabilex sheets 0.125 mm thick). Three impressions were taken of it with each of the following: the polyvinylsiloxane Examix-GC-(AdEx), Aquasil-Dentsply-(AdAq) and Panasil-Kettenbach-(AdPa), and the polydimethylsiloxane Densell-Dental Medrano-(CoDe), Speedex-Coltene-(CoSp) and Lastic-Kettenbach-(CoLa). All impressions were taken with putty and light-body materials using a one-step technique. Standardized digital photographs were taken at different time intervals (0, 15, 30, 60, 120 minutes; 24 hours; 7 and 14 days), using an "ad-hoc" device, and analyzed using software (Image Tool) by measuring the distance between lines previously made at the top of the mold. The results were analyzed by ANOVA for repeated measures. The initial and final values for mean and SD were: AdEx: 1.32 (0.01) and 1.31 (0.00); AdAq: 1.32 (0.00) and 1.32 (0.00), AdPa: 1.327 (0.006) and 1.31 (0.00); CoDe: 1.32 (0.00) and 1.32 (0.01); CoSp: 1.327 (0.006) and 1.31 (0.00), CoLa: 1.327 (0.006) and 1.303 (0.006). Statistical evaluation showed that both material and time have significant effects., Conclusion: Under the conditions in this study we conclude that time would significantly affect the lineal dimensional stability of elastomeric impression materials.

Progressive lung damage in cystic fibrosis (CF) has been linked to inadequate airway mucosal hydration. We previously demonstrated that an inositol tetrakisphosphate analog, 1-O-octyl-2-O-butyryl-myo-inositol 3,4,5,6-tetrakisphosphate octakis(propionoxymethyl)ester (INO-4995), regulates airway secretory and absorptive processes, affecting mucosal hydration by prolonged (24 h) inhibition of Na(+) and fluid absorption in CF human nasal epithelia (CFHNE). The objectives of this study were to further assess clinical potential of INO-4995 in CF through ascertaining in vivo activity in mice with CF, determining the effects of repeated administration on potency and determining cytoplasmic half-life. Uptake and metabolism of [(3)H]INO-4995 was monitored with HPLC to calculate intracellular half-life. INO-4995 was administered in vitro repeatedly over 4 to 8 days to CFHNE. Fluid absorption was assessed by blue dextran exclusion, and basal short-circuit current was measured in Ussing chambers. INO-4995 (1-100 microg/kg) was dosed intranasally either as a single dose or once per day over 4 days to gut-corrected CF mice. [(3)H]INO-4995 was rapidly taken up by epithelial cultures and converted to the active drug, which had a half-life of 40 hours. Repeated daily application of INO-4995 to CFHNE lowered the effective concentration for inhibition of fluid absorption and amiloride-sensitive short-circuit current in cultured CFHNE, and reduced nasal potential difference to nearly control levels in gut-corrected CF mice. Ca(2+)-activated Cl(-) channel activity was also boosted in cultures. Mouse nasal levels fell from abnormal levels to within 2 muA of normal with repeated exposure to 0.8 microg/kg over 4 days. These data support further development of INO-4995 for the treatment of CF.

The aim of this study was to evaluate the clinical behavior of two conventional glass ionomers used for pit and fissure sealing in terms of retention, marginal adaptation, caries recurrence and cracking. Eighty-three fully erupted first permanent molars were sealed, in a group of children aged 5 to 8 years. A double-blind, single operator, paired design was used. Materials applied were Fuji IX and VII. Retention (R), presence of caries (PC), marginal discoloration (MD), marginal adaptation (MA) and cracking (C) were evaluated at 6 and 12 months using Ryge's criteria. Data registered 6 months after treatment were: Fuji IX: R: Alpha 37, Bravo 2, Charlie: 4; PC: Alpha 43, Bravo: 0; MA: Alpha 38, Bravo: 1; MD: Alpha 39, Bravo: 0, Charlie: 0; C: Alpha 39, Bravo: 0. Fuji VII: R: Alpha 29, Bravo 4, Charlie: 7; PC: Alpha 40, Bravo: 0; MA: Alpha: 34, Bravo: 0; MD: Alpha: 34, Bravo: 0, Charlie: 0; C: Alpha 33, Bravo: 0. Results after one year were: Fuji IX: R: Alpha 33, Bravo: 3, Charlie: 9; PC: Alpha 44, Bravo: 1; MA: Alpha 33, Bravo: 1; MD: Alpha: 34, Bravo: 0, Charlie: 0; C: Alpha: 34, Bravo: 0. Fuji VII: R: Alpha 22, Bravo 4, Charlie: 13, PC: Alpha 40, Bravo: 0; MA: Alpha 23, Bravo: 1; MD: Alpha 23, Bravo: 1, Charlie: 0; C: Alpha 23, Bravo: 1. Statistical analysis using Fisher test showed no significant difference (p > 0.05) for R, and MA. For PC, MD and C, values are not reported because both materials showed the same results. The second control showed no significant difference (p > 0.05) for R, MA, PC, MD and C. Results suggest no difference between Fuji IX and Fuji VII as sealants in fully erupted permanent molars.

An ion, atom, molecule or macro-particle in a trap can exhibit large motional oscillations due to the Doppler-affected radiation pressure by a laser, blue-detuned from an absorption line of a particle. This oscillator can be nearly thresholdless, but under certain conditions it may exhibit huge hysteretic excitation. Feasible applications include a "Foucault pendulum" in a trap, a rotation sensor, single atom spectroscopy, isotope separation, etc.

Relativistically-intense laser beam with large field gradient ("laser gate") enables strong inelastic scattering of electrons crossing the beam. This process allows for multi-MeV electron net acceleration per pass within the wavelength space. Inelastic scattering even in low-gradient laser field may also induce extremely tight temporal focusing and electron bunch formation down to quantum, zeptosecond limit.

We predict nanoscale field and dipole patterns due to the broken uniformity of a laser-driven local field in 1D and 2D lattices. They may result in size-related resonances and large field enhancement, which in turn can give rise to low-intensity nonlinear optical effects, e.g., optical bistability, even in the ultimate case of a pair of coupled atoms. At certain, "magic" numbers and configurations of atoms in a lattice, the system may exhibit the self-induced cancellation of the suppression of a local field.

The aim of this study was to evaluate in vitro the duration of the antimicrobial effect of endodontic sealers by means of the Direct Contact Test. The sealers tested were: Endomethasone - Septodont, Endomethasone C-Septodont, Endion-Voco, Diaket-ESPE, Pulp Canal Sealer-SybronEndo, and AH26-Dentsply DeTrey. The endodontopathic microorganisms (MO) confronted were: Staphylococcus aureus (Sa), Candida albicans (Ca), Enterococcus faecalis (Ef), Prevotella intermedia (Pi), Porphyromonas gingivalis (Pg) and Fusobacterium nucleatum (Fn). Test specimens of each sealer were prepared and placed on the surface of agar plates that had been inoculated with each MO, and after predetermined periods, transfers were made from the contact area between the test specimen and the cultured agar and from the area that had not been in contact with the test specimens (control). The results were read as presence/absence of microbial growth and analyzed statistically using the Kruskal-Wallis test. It was concluded that the structural features and virulence of endodontopathic microorganisms determine their response to the sealers, independently of the time during which sealers act and the mechanism by which the antiseptic reaches the microorganism, which in this case was by direct contact.

We propose a free-space terminator and broadband interferometry based on a coherent blackbody effect in an ultrathin nonreflecting metallic layer in the microwave to infrared domains. A frequency-insensitive device consisting of a metallic layer in a ring (Sagnac) interferometer can be used for autocorrelation measurements of extremely broad EM spectra.

Amiloride-sensitive, epithelial Na(+) channel (ENaC)-mediated, active absorption of Na(+) is elevated in the airway epithelium of cystic fibrosis (CF) patients, resulting in excess fluid removal from the airway lumen. This excess fluid/volume absorption corresponds to CF transmembrane regulator-linked defects in ENaC regulation, resulting in the reduced mucociliary clearance found in CF airways. Herein we show that INO-4995, a synthetic analog of the intracellular signaling molecule, D-myo-inositol 3,4,5,6-tetrakisphosphate, inhibits Na(+) and fluid absorption across CF airway epithelia, thus alleviating this critical pathology. This conclusion was based on electrophysiological studies, fluid absorption, and (22)Na(+) flux measurements in CF airway epithelia, contrasted with normal epithelia, and on electrophysiological studies in Madin-Darby canine kidney cells and 3T3 cells overexpressing ENaC. The effects of INO-4995 were long-lasting, dose-dependent, and more pronounced in epithelia from CF patients vs. controls. These findings support preclinical development of INO-4995 for CF treatment and demonstrate for the first time the therapeutic potential of inositol polyphosphate derivatives.

A relativistic field-gradient (ponderomotive) force in a laser standing wave ceases to exist in a familiar form; e.g., the adiabatic Hamiltonian is not separable into kinetic and potential energies for electrons moving in the antinode planes. We show that the force in the direction across the initial motion of an electron reverses its sign and makes the high-field areas attractive for electrons, opposite to a regular ponderomotive force. The reversal occurs at a relativistic-scale incident momentum, and represents the only effect known so far that pins down a distinct borderline between relativistic and nonrelativistic motion.

Unlabelled: The aim of this study was to evaluate the antimicrobial activity of points containing antiseptics used for temporary obturation in endodontics. Points containing calcium hydroxide (Roeko and Hygienic), and chlorhexidine (Roeko) were tested and gutta-percha points served as control (Meta Dental Corp.). The following microorganisms were studied: Streptococcus mutans (Sm), Enterococcus faecalis (Ef), Staphylococcus aureus (Sta), Candida albicans (Ca), Fusobacterium nucleatum (Fn) ATCC 25586, Porphyromonas gingivalis (Pg) ATCC 33277, and Prevotella intermedia (Pi) ATCC 25611. The experiments were organized in three stages: 1st stage: one point of each kind was placed on agar plates previously seeded with microorganisms. 2nd stage: Another group of points was immersed in broth for 24 hours and placed on the seeded agar. Samples were then incubated at 37 degrees in the conditions of oxygen and for the time required by each microorganism. The zones of bacterial inhibition around each point were measured. The pH values of the broths were recorded. 3rd stage: the immersion broths were inoculated with suspensions of microorganisms, incubated, seeded on plates by dissemination and incubated. CFU counts were performed., Results: points containing chlorhexidine showed inhibition zones with every microorganism in the 1st stage and, in the 2nd stage, with most of the microorganisms studied, except for Fn and Pi. Calcium hydroxide containing points did not inhibit any of the microorganisms assessed. Broth pH values did not exhibit any changes. CFU counts of the broths in which chlorhexidine points had been immersed showed total inhibition for all the microorganisms. The differences between materials were statistically significant (p<0.05) (ANOVA). In the conditions of this study, chlorhexidine-containing points proved to be effective against most of the tested strains.

Conventional glass-ionomer cements (GICs) have a slow maturation time. Reduction in time of maturation may be achieved by acceleration of the setting reaction. One factor that assists this is the reduction in glass particle size producing a larger surface area for reaction. The resulting rapid set and more rapid maturation should potentially lead to less long-term degradation. Biaxial strength measurements were made with respect to time for two GICs of similar compositions but with differing particle size distributions at different time intervals after immersion in both water and artificial saliva. There was little difference between the strength of the two materials over periods up to 12 weeks. A theoretical estimation of the relative surface areas of glasses showed that, despite there being twice the surface area available for reaction for one glass, there was little difference in strengths values between the two materials at any of the times tested here. The similarity in strength values despite this substantial difference suggests that the larger particles may have a greater influence in the cement forming process.

We predict that Coulomb explosion of a nanoscale cluster, which is ionized by high-intensity laser radiation and has a naturally occurring spatial density profile, will invariably produce shock waves. In most typical situations, two shocks, a leading and a trailing one, form a shock shell that eventually encompasses the entire cluster. Being the first example of shock waves on the nanometer scale, this phenomenon promises interesting effects and applications, including high-rate nuclear reactions inside each individual cluster.

Aim: To determine the flow characteristics and subcutaneous tissue reactions to five endodontic sealers., Methodology: The materials used were Procosol, AH26, Endomethasone, Sealapex and Endion. The sealers were prepared following the manufacturers' instructions, and 0.075 mL of each material was placed on a glass surface, which was then rotated 90 degrees. The samples were stored at 37 degrees C and 95% humidity. The displacement of the sealer was recorded by measuring the difference between its original position and the position recorded at 15 and 60 min. Three samples of each material were used. Two pockets were created in the back of Wistar rats, and one silicone tube, 1 mm in diameter and 1 cm in length, was implanted in each. One was filled with one of the materials under study, and the other empty tube was implanted as a control. Fourteen days after implantation, the animals were sacrificed, and samples of the skin containing the tubes were histologically processed. Histological and histomorphometric evaluations of the tissues adjacent to the open end of the tube were carried out the volume of tissue reaction was measured histomorphometrically according to standard stereological principles. Results were statistically analysed using analysis of variance and Duncan's test., Results: The highest flow values were obtained with Sealapex and AH26. Time significantly affected the flow and the material (P < (1001). Procosol and Endion produced the most severe histological reactions: these were outlined by fibrous tissue; AH26. Endomethasone and Sealapex produced reactions of smaller size and with more moderately defined limits., Conclusions: The flow did not correlate with the degree of inflammatory response. Procosol and Endion produced the most severe tissue reactions, whereas Endomethasone, Sealapex and AH26 produced only minimum reactions.

Cystic fibrosis (CF) patients suffer from a defect in hydration of mucosal membranes due to mutations in the cystic fibrosis transmembrane regulator (CFTR), an apical chloride channel in mucosal epithelia. Disease expression in CF knockout mice is organ specific, varying with the level of expression of calcium activated Cl(-) channels (CLCA). Therefore, restoring transepithelial Cl(-) secretion by augmenting alternate Cl(-) channels, such as CLCA, could be beneficial. However, CLCA-mediated Cl(-) secretion is transient, due in part to the inhibitory effects of myo-inositol 3,4,5,6-tetrakisphosphate [Ins(3,4,5,6)P(4)]. This suggests that antagonists of Ins(3,4,5,6)P(4) could be useful in treatment of CF. We have, therefore, synthesized a series of membrane-permeant Ins(3,4,5,6)P(4) derivatives, carrying alkyl substituents on the hydroxyl groups and screened them for effects on Cl(-) secretion in a human colonic epithelial cell line, T(84). While membrane-permeant Ins(3,4,5,6)P(4) derivatives had no direct effects on carbachol-stimulated Cl(-) secretion, Ins(3,4,5,6)P(4) derivatives, but not enantiomeric Ins(1,4,5,6)P(4) derivatives, reversed the inhibitory effect of Ins(3,4,5,6)P(4) on subsequent thapsigargin activation of Cl(-) secretion. The extent of the antagonistic effect of the Ins(3,4,5,6)P(4) derivatives varied with the position of the alkyl substituents. Derivatives with a cyclohexylidene ketal or a butyl-chain at the 1-position reversed the Ins(3,4,5,6)P(4)-mediated inhibition of Cl(-) secretion by up to 96 and 85%, respectively, whereas butylation of the 1- and 2-position generated a reversal effect of only 65%. Derivatives carrying the butyl chain only at the 2-position showed no antagonistic effect. These data: (1) Support the hypothesis that Ins(3,4,5,6)P(4) stereospecifically inhibits Ca(2+) activated Cl(-) secretion and that Ins(3,4,5,6)P(4) mediates most, if not all of the cholinergic-mediated inhibition of chloride secretion in T(84) cells; (2) Demonstrate Ins(3,4,5,6)P(4)-mediated inhibition can be completely reversed with rationally designed membrane-permeant Ins(3,4,5,6)P(4) antagonists; (3) Demonstrate that a SAR for membrane-permeant Ins(3,4,5,6) P(4) antagonists can be generated and screened in a physiologically relevant cell-based assay; (4) Indicate that Ins(3,4,5,6)P(4) derivatives could serve as a starting point for the development of therapeutics to treat cystic fibrosis.

Electromagnetic bursts of substantial energy on a nuclear time scale of 10(-21)-10(-22) s [zeptosecond (zs) to sub-zs] can theoretically be generated by a perawatt or multiterawatt laser beam focused on a subwavelength-size solid particle or thin wire. Terawatt laser in a similar setup could be instrumental in reaching the subattosecond domain. The system may also generate a half-cycle pulse magnetic field on astrophysical scale up to approximately 10(6) T.

When evaluating the antimicrobial effect of endodontic sealers it is important to study the variation of pH, since this is one of the properties that may inhibit the microbial growth and influence biocompatibility. The aim of this study was to evaluate the capacity of several endodontic sealers without calcium hydroxide to modify the pH of the solution in which they were immersed. The sealers used were: Endomethasone C-Septodont (EC), Bio Seal-Ogna (BS), Diaket-Espe (D), Procosol-StarDental (P), AH26-Dentsply (AH), Ketac Endo-Espe (KE), AHPlus-Dentsply (AHP), Endion-Voco (E), Endomethasone-Septodont (EM), EZ Fill-Essential Dental Systems (EZ). Samples of each material were placed in tubes with phosphate saline solution (PBS). The pH was determined at 2, 5, 12, 20, 30, 60, and 90 days. A tube with solution alone was taken as control. Statistical analysis of the results was performed by ANOVA. Material, sample, time and material-time interaction elicited statistically significant differences (p < 0.05). The variation of pH produced by immersion of endodontic sealers depends on the composition of the material.

The superoxide-generating neutrophil NADPH oxidase can be activated in cell-free reconstitution systems by several agonists, most notably arachidonic acid and the detergent sodium dodecyl sulfate. In this study, we show that both phosphatidic acids and diacylglycerols can serve separately as potent, physiologic activators of NADPH oxidase in a cell-free system. Stimulation of superoxide generation by these lipids was dependent upon both Mg(2+) and agonist concentration. Activation of NADPH oxidase by phosphatidic acids did not appear to require their conversion to corresponding diacylglycerols by phosphatidate phosphohydrolase, since diacylglycerols were much slower than phosphatidic acids to activate the system and required the presence of ATP. Stimulation of the oxidase by dioctanoylglycerol proved to be by a means other than the activation of protein kinase C. Instead, dioctanoylglycerol was converted to dioctanoylphosphatidic acid by an endogenous diacylglycerol kinase present in the cell-free reaction system. This conversion was sensitive to the diacylglycerol kinase inhibitor R59949 and explains the markedly slower kinetics of activation and the novel ATP requirement seen with dioctanoylglycerol. The level of dioctanoylphosphatidic acid formed was suboptimal for NADPH oxidase activation but could synergize with the unmetabolized dioctanoylglycerol to activate superoxide generation.

The aim of this study was to determine the in vitro antimicrobial effect of six endodontic sealers after 2, 20 and 40 days. The sealers studied were Apexit, Endion, AH26, AH-Plus. Procosol and Ketac Endo. The microorganisms used were Candida albicans, Staphylococcus aureus, Streptococcus mutans and Actinomyces israelii. Petri dishes were filled with sterile agar and 0.1-ml wells were prepared and filled with the sealers. The agar plates were stored for 24 h at 37 degrees C. The samples were then removed, immersed in 4.5 ml of culture medium and divided into three groups. The samples in group 1 were stored for 2 days at 37 degrees C whereas the samples of groups 2 and 3 were stored at 4 degrees C for 20 and 40 days respectively. The samples were then removed and discarded, and 0.1 ml of the culture medium was seeded on the agar plates in order to perform colony forming unit counts. Apexit, Endion and AH-Plus produced slight inhibition on Streptococcus mutans at 20 days and on Actinomyces israelii at every time interval. No effect was found on Candida albicans and Staphylococcus aureus. Ketac Endo only produced an antimicrobial effect on Actinomyces israelii at 2 and 40 days. AH26 and Procosol showed antimicrobial effect at 40 days on Candida albicans, at 20 and 40 days on Streptococcus mutans and Staphylococcus aureus, and an effective inhibition on Actinomyces israelii at every time interval. Statistical analysis revealed both sealers and microorganisms to be significant factors affecting results in groups 2 and 3. In conclusion, the sealers evaluated in this study showed different inhibitory effects depending on time span. Overall, sealers containing cugenol and formaldehyde proved to be most effective against the microorganisms at the time intervals studied.

We have synthesized the first deoxy analogues of myo-inositol 3,4,5, 6-tetrakisphosphate (1) [Ins(3,4,5,6)P4], rac-2-deoxy-myo-inositol 3, 4,5,6-tetrakisphosphate (rac-2), 2-deoxy-myo-inositol 1,4,5, 6-tetrakisphosphate (ent-2), and rac-1-deoxy-myo-inositol 3,4,5, 6-tetrakisphosphate (rac-3). In order to evaluate the binding properties of the three derivatives to the yet unidentified intracellular binding sites for Ins(3,4,5,6)P4, the analogues were converted to membrane-permeant derivatives. Starting with common inositol precursors, various forms of Barton-McCombie deoxygenation and classical protection/deprotection procedures yielded the desired precursors rac-1-O-butyryl-2-deoxy-myo-inositol (rac-12), ent-3-O-butyryl-2-deoxy-myo-inositol (ent-12), and rac-2-O-butyryl-1-deoxy-myo-inositol (rac-19), respectively. Phosphorylation and subsequent deprotection yielded rac-2, ent-2, and rac-3. Alternatively, phosphorylation followed by alkylation with acetoxymethyl bromide gave the membrane-permeant derivatives 1-O-butyryl-2-deoxy-myo-inositol 3,4,5,6-tetrakisphosphate octakis(acetoxymethyl) ester (rac-5), 3-O-butyryl-2-deoxy-myo-inositol 1,4,5,6-tetrakisphosphate octakis(acetoxymethyl) ester (ent-5), and 2-O-butyryl-1-deoxy-myo-inositol 3,4,5,6-tetrakisphosphate octakis(acetoxymethyl) ester (rac-6), respectively. We examined the potency of the membrane-permeant deoxy derivatives in inhibition of calcium-mediated chloride secretion (CaMCS) in intact T84 cells. Compared to the 1,2-di-O-butyryl-myo-inositol 3,4,5, 6-tetrakisphosphate octakis(acetoxymethyl) ester (4), the membrane-permeant derivative of Ins(3,4,5,6)P4 (1), the 2-deoxy derivative (rac-5) exhibited a slightly weaker inhibitory effect, while the enantiomerically pure 2-deoxy-Ins(1,4,5,6)P4 (ent-5) and the 1-deoxy derivative (rac-6) were inactive. As expected, the effect was stereoselective. Thus, the 1-hydroxyl group is apparently essential for binding and the inhibitory effect of Ins(3,4,5,6)P4 on chloride secretion, whereas the 2-hydroxyl group plays a less important role.

The synthesis of rac-2,5,6-tri-O-butyryl-myo-inositol 1,3,4-trisphosphate hexakis(acetoxymethyl) ester [Bt3-Ins(1,3,4)P3/AM, 1], a membrane-permeant derivative of myo-inositol 1,3,4-trisphosphate [Ins(1,3,4)P3] is reported. 1 inhibited calcium-mediated chloride secretion of T84 cells, suggesting a regulatory link of Ins(1,3,4)P3 and the biosynthesis of the known inhibitor myo-inositol 3,4,5,6-tetrakisphosphate.

To understand how phosphatidylinositol 3-kinase (PI3K) modulates cell structure and function, we examined the molecular and cellular defects of a Dictyostelium mutant strain (pik1(Delta)2(Delta)) missing two (DdPIK1 and 2) of three PI3K genes, which are homologues of the mammalian p110 subunit. Levels of [32P]phosphatidylinositol 3, 4 bisphosphate (PI(3,4)P2) and [32P]phosphatidylinositol trisphosphate (PIP3) were reduced in pik1(Delta)2(Delta), which had major defects in morphological and functional correlates of macropinocytosis. This was accompanied by dramatic deficits in a subset of F-actin-enriched structures such as circular ruffles, actin crowns and pseudopodia. Although pik1(Delta)2(Delta) were mobile, they failed to aggregate into streams. Therefore we conclude that PIK1 and 2, possibly through modulation of the levels of PIP3 and PI(3,4)P2, regulate the organization of actin filaments necessary for circular ruffling during macropinocytosis, the extension of pseudopodia and the aggregation of cells into streams, but not the regulation of cell motility.

Several inositol-containing compounds play key roles in receptor-mediated cell signaling events. Here, we describe a function for a specific inositol polyphosphate, D-myo-inositol 1,4,5,6-tetrakisphosphate [Ins(1,4,5,6)P4], that is produced acutely in response to a receptor-independent process. Thus, infection of intestinal epithelial cells with the enteric pathogen Salmonella, but not with other invasive bacteria, induced a multifold increase in Ins(1,4,5,6)P4 levels. To define a specific function of Ins(1,4,5,6)P4, a membrane-permeant, hydrolyzable ester was used to deliver it to the intracellular compartment, where it antagonized epidermal growth factor (EGF)-induced inhibition of calcium-mediated chloride (Cl-) secretion (CaMCS) in intestinal epithelia. This EGF function is likely mediated through a phosphoinositide 3-kinase (PtdIns3K)-dependent mechanism because the EGF effects are abolished by wortmannin, and three different membrane-permeant esters of the PtdIns3K product phosphatidylinositol 3,4,5-trisphosphate mimicked the EGF effect on CaMCS. We further demonstrate that Ins(1,4,5,6)P4 antagonized EGF signaling downstream of PtdIns3K because Ins(1,4,5, 6)P4 interfered with the PtdInsP3 effect on CaMCS without affecting PtdIns3K activity. Thus, elevation of Ins(1,4,5,6)P4 in Salmonella-infected epithelia may promote Cl- flux by antagonizing EGF inhibition mediated through PtdIns3K and PtdInsP3.

The disintegration of three endodontic cements in water was determined quantitatively and qualitatively. The materials studied were Ketac-Endo (KE), Tubli Seal (TS), and AH26 (AH). Specimens were immersed in water for 48 h (GI), 7 (GII) and 45 days (GIII). The solid residue was then determined. For the qualitative analysis three groups of tubes were filled with the materials and stored in water for the same periods. The exposed surface was photographed. Results expressed as percentage of original mass in the quantitative analysis for loss of mass due to dissolution were: GI = KE 2.39 (0.70); TS 3.56 (0.37); AH 4.94 (2.83); GII = KE 2.84 (0.30); TS 2.50 (0.50); AH 0.66 (0.26); GIII = KE 1.60 (0.84); TS 1.03 (0.42); AH 1.22 (0.54). Tukey's least significant difference (0.05) was 2.94. In the qualitative experiment KE disintegration was far more evident than that suffered by other materials. The quantitative results had no correlation with the qualitative observations probably due to the difference in the moment when the materials were immersed.

Electromagnetic (EM) bubbles (EMB's), unipolar, super-short, and intense nonoscillating solitary pulses of EM radiation, can be generated in a gas of nonlinear atoms by available half-cycle pulses (HCP's). We investigate how EMB's characteristics (amplitude, length, formation distance, and total number) are controlled by the amplitude and length of originating HCP's. We also predict shocklike wave fronts in the multibubble regime.

T84 monolayers were studied to determine the effect of AlF4, an activator of heterotrimeric G proteins, on Cl-secretion and intestinal barrier function. Basolateral (but not apical) addition of AlF4- increased short-circuit current (I(sc)) and decreased transepithelial resistance. Preincubation with the heavy metal chelator deferoxamine showed that both effects were dependent on Al3+. The effect on I(sc) was abolished by the intracellular Ca2+ chelator 1,2-bis(2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid or in Cl(-)-free solutions, whereas the decrease in resistance was unaffected. AlF4- also increased intracellular Ca2+, as assessed via fura 2 fluorometry. AlF4- had no effect on adenosine 3',5'-cyclic monophosphate (cAMP) or guanosine 3',5'-cyclic monophosphate (cGMP) levels in T84 cells. The effect of AlF4- on transepithelial resistance was accompanied by a decrease in cellular F-actin as well as increased transepithelial fluxes of the paracellular markers mannitol and inulin. The results indicate that AlF4(-)-sensitive G proteins regulate both epithelial secretory and barrier functions, but via different pathways. AlF4- increases Cl- secretion via a Ca2+-dependent and cAMP- and cGMP-independent mechanism in T84 cells, whereas the decrease in resistance is independent of Ca2+.

Epidermal growth factor (EGF) and carbachol both inhibit calcium-activated chloride secretion by the human colonic epithelial cell line, T84. Although the inhibitory mechanism for the carbachol effect involves the 3,4,5,6-isomer of inositol tetrakisphosphate, the mechanisms responsible for the EGF effect have not yet been fully elucidated. Here, we studied the role of phosphatidylinositol 3-kinase (PI 3-kinase) in the inhibitory effect of EGF. The PI 3-kinase inhibitor, wortmannin, slightly increased basal chloride secretion and potentiated the secretory response to thapsigargin. Wortmannin also partially reversed EGF-induced, but not carbachol-induced, inhibition of thapsigargin-stimulated chloride secretion. Wortmannin alone had no effect on carbachol- or histamine-induced chloride secretion and completely reversed EGF-induced inhibition of the secretory response to these agonists. EGF, carbachol, histamine, and thapsigargin all increased levels of the 85-kDa regulatory subunit of PI 3-kinase in antiphosphotyrosine immunoprecipitates. However, only EGF significantly increased levels of the 110-kDa catalytic subunit. Furthermore, only EGF increased PI 3-kinase activity in an in vitro kinase assay. High levels of phosphatidylinositol (3)-monophosphate were present in unstimulated cells and significantly reduced by wortmannin. EGF, but not carbachol, rapidly increased levels of phosphatidylinositol (3,4)-bisphosphate and phosphatidylinositol (3,4,5)-trisphosphate. Production of these lipids was also sensitive to wortmannin. Our data suggest that EGF activates PI 3-kinase and that its lipid products may mediate the inhibitory effect of EGF on calcium-dependent chloride secretion. Our data also suggest that a phosphatidylinositol-specific 3-kinase activity is present in unstimulated T84 cells and may regulate production of phosphatidylinositol (3)-monophosphate and basal secretory tone.

The signaling pathway leading from G protein-coupled chemoattractant receptors to the generation of oxidants by NADPH oxidase in human neutrophils requires the formation of the lipid mediator phosphatidylinositol 3,4,5-trisphosphate (PIP3). Two mechanisms through which PIP3 can be generated have been described in human leukocytes. One pathway involves the coupling of the src-related tyrosine kinase Lyn to the "classical" p85/p110 form of phosphatidylinositol 3-kinase. The second paradigm utilizes a novel form of phosphatidylinositol 3-kinase whose activity is directly regulated by G protein betagamma subunits. In this paper, we show that formation of PIP3 in chemoattractant-stimulated neutrophils is substantially attenuated by inhibitors that specifically block tyrosine kinase activity. These data suggest that the Lyn activation pathway plays a major role in the formation of this important lipid messenger during chemoattractant stimulation of human neutrophils.

This study examined whether epidermal growth factor (EGF) inhibits Ca(2+)-dependent Cl- secretion by T84 cells. Basolateral EGF inhibited Cl- secretion induced by carbachol or thapsigargin, without blocking the rise in intracellular Ca2+. Studies have shown that carbachol renders T84 cells refractory to subsequent stimulation by thapsigargin, an effect ascribed to D-myo-inositol 3,4,5,6-tetrakisphosphate [D-Ins(3,4,5,6)P4]. EGF also increased DL-Ins(3,4,5,6)P4 to a maximum of 170% above control. However, despite the fact that EGF inhibited Cl- secretion at 1 min, DL-Ins(3,4,5,6)P4 was not elevated at this time point. EGF plus carbachol had a greater inhibitory effect on Cl- secretion than either alone, indicating the likely involvement of an additional inhibitory pathway activated by EGF. Staurosporine did not alter the ability of EGF to inhibit Cl- secretion or increase DL-Ins(3,4,5,6)P4. In contrast, genistein inhibited the rise in DL-Ins(3,4,5,6)P4 and partially reversed EGF's inhibitory effect on Cl- secretion. In conclusion, EGF and carbachol can both inhibit Cl- secretion via D-Ins(3,4,5,6)P4, whereas EGF also generates an additional inhibitory signal.

The Rac GTP-binding proteins are members of the Rho family and regulate growth factor-stimulated actin assembly in a variety of cells. The formation of phosphorylated inositol lipids has been implicated in control of the processes initiating and regulating such actin polymerization. Associations of Rho family GTP-binding proteins with enzymes involved in lipid metabolism have been described. Here we demonstrate a direct and specific interaction of Rac proteins with phosphatidylinositol (PI) 3-kinase. This interaction is dependent upon Rac being in a GTP-bound state and requires an intact Rac effector domain. In contrast, direct binding of RhoA to PI 3-kinase could not be detected. Rac-GTP also bound to PI 3-kinase in Swiss 3T3 fibroblast and human neutrophil lysates, and increased PI 3-kinase activity became associated with Rac-GTP in platelet-derived growth factor-stimulated cells. Interaction of Rac-GTP with PI 3-kinase in vitro stimulated the activity of the enzyme by 2-9-fold. A specific interaction of active Rac with PI 3-kinase might be important in regulation of the actin cytoskeleton.

The aim of the present study was to analyze different substrates used to determine composite-dentine bond strength. It comprises 3 parts. In the first, results obtained using bovine and human dentin are compared. In the second, bond strength of composite-bovine dentin sections cut at different angles was studied. In the third part, the effect of different storage solutions on bond strength was analysed. Two methods were used to study the capacity of bovine dentin to reproduce "in vivo" conditions as closely as possible: a) bond strength tests. Tensile strength of composite samples mounted on flat dentin surfaces, treated with bonding agents, stored in artificial saliva at 37 degrees C was measured. b) Observation of exposed surfaces by scanning electron microscopy. Results showed that more reliable values were obtained when using bovine dentin stored in distilled water at 4 degrees C sectioned at an angle of 125 degrees to vestibular face and not further than 7 mm from the incisal edge.