Effects of flowable liners on the shrinkage vectors of bulk-fill composites.

OBJECTIVES: This investigation evaluated the effect of flowable liners beneath a composite restoration applied via different methods on the pattern of shrinkage vectors. METHODS: Forty molars were divided into five groups (n = 8), and cylindrical cavities were prepared and bonded with a self-etch adhesive (AdheSe). Tetric EvoCeram Bulk Fill (TBF) was used as the filling material in all cavities. The flowable liners Tetric EvoFlow Bulk Fill (TEF) and SDR were used to line the cavity floor. In gp1-TBF, the flowable composite was not used. TEF was applied in a thin layer in gp2-fl/TEF + TBF and gp3-fl/TEF + TBFincremental. Two flowable composites with a layer thickness of 2 mm were compared in gp4-fl/TEF + TBF and gp5-fl/SDR + TBF. TEF and SDR were mixed with radiolucent glass beads, while air bubbles inherently present in TBF served as markers. Each material application was scanned twice by micro-computed tomography before and after light curing. Scans were subjected to image segmentation for calculation of the shrinkage vectors. RESULTS: The absence of a flowable liner resulted in the greatest shrinkage vectors. A thin flowable liner (gp2-fl/TEF + TBFbulk) resulted in larger overall shrinkage vectors for the whole restoration than a thick flowable liner (gp4-fl/TEF + TBF). A thin flowable liner and incremental application (gp3-fl/TEF + TBFincremental) yielded the smallest shrinkage vectors. SDR yielded slightly smaller shrinkage vectors for the whole restoration than that observed in gp4-fl/TEF + TBF. CONCLUSIONS: Thick flowable liner layers had a more pronounced stress-relieving effect than thin layers regardless of the flowable liner type. CLINICAL RELEVANCE: It is recommended to apply a flowable liner (thin or thick) beneath bulk-fill composites, preferably incrementally.

Transillumination and HDR Imaging for Proximal Caries Detection.

The purpose of this study was to develop an in vitro model for the validation of near-infrared transillumination (NIRT) for proximal caries detection, to enhance NIRT with high-dynamic-range imaging (HDRI), and to compare both methods, using micro-computed tomography (µCT) as a reference standard. Both proximal surfaces of 53 healthy or decayed permanent human teeth were examined using the Diagnocam (DC) (KaVo) and NIRT with HDRI (NIRT-HDRI). NIRT was combined with HDRI to improve the diagnostic performance by reducing under- and overexposed image areas. For NIRT-HDRI, an exposure series was captured and merged into a single HDR image. A classification was applied according to lesion depth. All surfaces were assessed twice by 2 trained examiners, and additionally with µCT for validation. The Kappa statistic was used to calculate inter-rater reliability and agreement between DC and NIRT-HDRI. Inter-rater reliability (weighted Kappa, wκ) showed very good agreement for the DC (0.90) and NIRT-HDRI (0.96). The overall agreement (wκ) was almost perfect (0.85). In the individual categories (0 to 4), the agreement (simple Kappa) ranged from almost perfect (category 4) to moderate (1 and 2) to substantial (categories 0 and 3). Sensitivity and specificity of sound surfaces, enamel, and dentin caries ranged from 0.57 to 0.99 and were similar for both methods in the different categories. NIRT-HDRI had a higher sensitivity for sound surfaces and enamel caries, as well as a higher specificity for dentin caries. Regarding the obtained images, HDRI allowed for the detection of caries within a greater range of luminance levels, resulting in a more detailed visualization of structures without under- or overexposure. However, HDRI this did not improve the diagnostics significantly. Distinguishing between a processed demineralized enamel and dentin lesions appears to be a problem specific to NIRT and cannot be balanced using HDRI.

Degradation Potential of Bulk Versus Incrementally Applied and Indirect Composites: Color, Microhardness, and Surface Deterioration.

This study investigated the color stability and microhardness of five composites exposed to four beverages with different pH values. Composite discs were produced (n=10); Filtek Z250 (3M ESPE) and Filtek P90 (3M ESPE) were applied in two layers (2 mm, 20 seconds), and Tetric N-Ceram Bulk Fill (TetricBF, Ivoclar Vivadent) and SonicFill (Kerr) were applied in bulk (4 mm) and then light cured (40 seconds, Ortholux-LED, 1600 mW/cm2). Indirect composite Sinfony (3M ESPE) was applied in two layers (2 mm) and cured (Visio system, 3M ESPE). The specimens were polished and tested for color stability; ΔE was calculated using spectrophotometer readings. Vickers microhardness (50 g, dwell time=45 seconds) was assessed on the top and bottom surfaces at baseline, 40 days of storage, subsequent repolishing, and 60 days of immersion in distilled water (pH=7.0), Coca-Cola (pH=2.3), orange juice (pH=3.75), or anise (pH=8.5) using scanning electron microscopy (SEM). The materials had similar ΔE values (40 days, p>0.05), but TetricBF had a significantly greater ΔE than P90 or SF (40 days). The ΔE was less for P90 and TetricBF than for Z250, SonicFill, and Sinfony (60 days). Repolishing and further immersion significantly affected the ΔE (p<0.05) except for P90. All composites had significantly different top vs bottom baseline microhardnesses. This was insignificant for the Z250/water, P90/orange juice (40 days), and Sinfony groups (40 and 60 days). Immersion produced variable time-dependent deterioration of microhardness in all groups. Multivariate repeated measures analysis of variance with post hoc Bonferroni tests were used to compare the results. ΔE and microhardness changes were significantly inversely correlated at 40 days, but this relationship was insignificant at 60 days (Pearson test). SEM showed degradation (40 days) that worsened (60 days). Bulk-fill composites differ regarding color-stability and top-to-bottom microhardness changes compared with those of other composites. P90 showed better surface degradation resistance. In conclusion, bulk-fill composites are not promising alternatives to incremental and indirect composites regarding biodegradation.

Anoctamin 6 is localized in the primary cilium of renal tubular cells and is involved in apoptosis-dependent cyst lumen formation.

Primary cilia are antenna-like structures projected from the apical surface of various mammalian cells including renal tubular cells. Functional or structural defects of the cilium lead to systemic disorders comprising polycystic kidneys as a key feature. Here we show that anoctamin 6 (ANO6), a member of the anoctamin chloride channel family, is localized in the primary cilium of renal epithelial cells in vitro and in vivo. ANO6 was not essential for cilia formation and had no effect on in vitro cyst expansion. However, knockdown of ANO6 impaired cyst lumen formation of MDCK cells in three-dimensional culture. In the absence of ANO6, apoptosis was reduced and epithelial cells were incompletely removed from the center of cell aggregates, which form in the early phase of cystogenesis. In line with these data, we show that ANO6 is highly expressed in apoptotic cyst epithelial cells of human polycystic kidneys. These data identify ANO6 as a cilium-associated protein and suggest its functional relevance in cyst formation.

Occlusal caries extension in relation to visual and radiographic diagnostic criteria: results from a microcomputed tomography study.

OBJECTIVE: This in vitro study aimed to evaluate occlusal caries extension in relation to visual and radiographic diagnostic criteria and their clinical value to indicate operative or preventive dental care. METHODS: A total of 196 third molars with clinically sound occlusal fissures or noncavitated lesions were collected. Before microcomputed tomography (µCT) investigation, each tooth was examined visually and radiographically. Kühnisch's µCT-based caries-extension index (CE index) was used to determine the caries depth on a numeric scale (0 = sound; 0.01-0.99 = enamel caries; 1.0-1.99 = dentin caries). Sensitivities (SEs), specificities (SPs), and area under the receiver operating characteristic curve (Az value) were also calculated. RESULTS: Based on µCT data, the following mean CE index values and standard deviations (SDs) were documented according to the visual criteria: sound = 0.6 (0.4); first visible signs = 0.9 (0.4); established lesions = 1.3 (0.3); microcavities = 1.4 (0.2); dentin exposure = 1.5 (0.2); and large cavities = 1.5 (0.3). The radiographic categories according to Marthaler (enamel caries [D0-2], caries in the outer half of dentin [D3], and caries in the inner half of dentin [D4]) were related to CE index values of 0.9 (0.4), 1.4 (0.2) and 1.6 (0.4), respectively. Caries detected visually or radiographically showed an SE of 84% and an SP of 85% (Az = 0.85). When both methods were used to predict dentin involvement simultaneously, SE = 27%, SP = 100%, and Az = 0.63; this combined visual and radiographic approach was associated with a perfect specificity and no false-negative decisions. The proportion of false-positive diagnoses was moderately high, and lesion extension in these cases was mainly limited to the outer 20% of the dentin. CONCLUSIONS: Our results might be useful for differentiating between preventive and operative dental care for pits and fissures.

Calcium-activated and apoptotic phospholipid scrambling induced by Ano6 can occur independently of Ano6 ion currents.

Immune cells and platelets maintain plasma membrane phospholipid asymmetry. Upon activation, this asymmetry is disrupted by phospholipid scrambling (PS), which is a major step during activation of immune cells, hemostasis and apoptosis. Anoctamin 6 (Ano6; TMEM16F) causes chloride (Cl(-)) and cation currents and is required for Ca(2+)-dependent PS. It is defective in blood cells from patients with Scott syndrome, a rare bleeding disorder. We examined if Cl(-) currents and PS are related, whether both processes are Ca(2+) dependent, and whether Ca(2+)-independent scrambling during intrinsic and extrinsic apoptosis is controlled by Ano6. Ca(2+) increase by ionomycin activated Ano6 Cl(-) currents and PS in normal lymphocytes, but not in B-lymphocytes from two different patients with Scott syndrome. Fas ligand (FasL) did not increase intracellular Ca(2+), but activated Cl(-) currents in normal but not in Scott lymphocytes. Whole-cell currents were inhibited by Cl(-) channel blockers and by siRNA knockdown of Ano6. In contrast, intrinsic mitochondrial apoptosis by ABT-737 did not induce Cl(-) currents in lymphocytes. PS was not inhibited by blockers of Ano6 or removal of Cl(-) ions. Remarkably, Ca(2+)-independent scrambling due to extrinsic (FasL) or intrinsic (ABT-737) apoptosis was unchanged in Scott cells. We conclude that: (i) Ano6 Cl(-) currents are activated by increase in cytosolic Ca(2+), or Ca(2+) independent by stimulation of Fas receptors; (ii) Ca(2+)-dependent PS induced by Ano6 does not require Cl(-) currents; (iii) Ca(2+)-independent PS does not require Ano6; (iv) Ano6 is necessary for Ca(2+)-dependent PS, but not by increasing intracellular Ca(2+).

Shape and volume of craniofacial cavities in intentional skull deformations.

Intentional cranial deformations (ICD) have been observed worldwide but are especially prevalent in preColombian cultures. The purpose of this study was to assess the consequences of ICD on three cranial cavities (intracranial cavity, orbits, and maxillary sinuses) and on cranial vault thickness, in order to screen for morphological changes due to the external constraints exerted by the deformation device. We acquired CT-scans for 39 deformed and 19 control skulls. We studied the thickness of the skull vault using qualitative and quantitative methods. We computed the volumes of the orbits, of the maxillary sinuses, and of the intracranial cavity using haptic-aided semi-automatic segmentation. We finally defined 3D distances and angles within orbits and maxillary sinuses based on 27 anatomical landmarks and measured these features on the 58 skulls. Our results show specific bone thickness patterns in some types of ICD, with localized thinning in regions subjected to increased pressure and thickening in other regions. Our findings confirm that volumes of the cranial cavities are not affected by ICDs but that the shapes of the orbits and of the maxillary sinuses are modified in circumferential deformations. We conclude that ICDs can modify the shape of the cranial cavities and the thickness of their walls but conserve their volumes. These results provide new insights into the morphological effects associated with ICDs and call for similar investigations in subjects with deformational plagiocephalies and craniosynostoses.

Primary cancellous bone formation with BMP and micro-chambered beads: experimental study on sheep.

PROBLEM: The physiological reconstruction of cancellous bone defects in surgery of the locomotor system is an unsatisfactorily solved problem. AIMS: The aims of this study are to examine whether micro-chambered ß-tricalcium-phosphate (ß-TCP) beads provide a certain capillary force suctioning in blood and bone marrow thus forming a stable "negative"-replica of the bone marrow spaces. If so, a new approach for osteoconduction would yield primarily a scaffold of lamellar cancellous bone under load without a long-lasting remodeling process. Recombinant human bone morphogenetic protein (rhBMP) might even enhance all processes of defect healing, remodeling and ß-TCP resorption; gentamicin-loaded ε-caprolactone might protect the implant. MATERIAL AND METHODS: Ten sheep were operated on; the patella-groove model and the tibial head were used. A defect of 9.4 × 20 mm was created using wet-grinding-diamond instruments. Micro-chambered ß-TCP-beads of 4-6 mm with 0.35 mg rhBMP-7 + 0.1 g collagen per animal, or 1.5 g demineralized bone matrix (DBM) paste on the contra-lateral side were implanted. Both osteoinduction groups were compared with the defect in the tibial heads where plain micro-chambered ceramic beads were inserted. Added to the beads was 12.5 mg gentamicin sulphate in 12.5 mg ε-caprolactone-carrier. Outward diffusion was prevented using a 1-mm-thick press-fit inserted ceramic lid. The bone healing, remodeling and resorption of the ceramic in a right-left comparison of the patella groove and the tibial head was examined at 6 weeks, 2 and 3 months; one animal in reserve was followed for 14 months. The animals were perfusion-fixed, the vasculature micro-casted with an acrylate and nondemineralized processed, and with µ-CT and microscopically documented. RESULTS: A primary load-bearing spongiosa had developed around the beads, which shortened the remodeling process. The strong micro-chambered, resorbable ß-TCP-beads demonstrate high capillary strength, resorb blood and bone marrow, and represent a stable formative material which, as a carrier for the controlled local release of BMP, that accelerates bone healing, shortens resorption and remodeling compared with plain and DBM loaded implants. CONCLUSION: Micro-chambered beads represent the bone-forming element, BMP yields a fast defect healing and enhanced remodeling of bone and resorption of ß-TCP compared to delayed and incomplete reconstruction and resorption of ß-TCP on the DBM-side, the plain implants reached nearly the same reconstruction, but far later compared with the BMP loaded implants.

Irregularities below the enamel-dentin junction may predispose for fissure caries.

This study investigated the structure of the fissure fundus on occlusal surfaces with respect to the detection of possible irregularities below the enamel-dentin junction (EDJ). Occlusal surfaces were examined by micro-computed tomography (µCT). In total, 203 third molars with clinically sound occlusal fissures or non-cavitated lesions were selected. All specimens were scanned with µCT. Subsequently, each tooth was sectioned, and each slice was investigated by stereomicroscopy. In 7 of 203 molars (3.4%), demarcated radiolucencies below the EDJ were detected by µCT. These defects were obviously of non-carious origin, because the µCT images revealed no gradient of demineralization in the dentin. In all cases, a direct pathway between the oral cavity and the dentin was evident. The comparison of the µCT sites with conventional histological images also revealed defects in the dentin. These results demonstrate that demarcated radiolucencies below the EDJ may not necessarily be caries lesions according to µCT images and may be classified as possible developmental irregularities. To avoid misinterpreting µCT data, dental researchers should carefully consider this condition when analyzing µCT images. The clinical significance of this finding is that these defects may predispose molar teeth to early-onset caries in occlusal pits and fissures.

Release of TEGDMA from composite during the chewing situation.

The aim of this study was to investigate the triethylene glycol (TEGDMA) elution kinetics from light-cured composite with and without chewing simulation over a time period of 86 h. An experimental composite with TEGDMA labeled with a tracer dose of 14C-TEGDMA was used. The material parameters were in the range of commercially available composites. The mastification was simulated with the Fatigue-machine and the MUC-3 chewing simulator. 14C was eluted to 2.55% of the applied 14C-TEGDMA dose within 86 h after chewing simulation with the Fatigue-machine and to 2.60% after chewing simulation with the MUC-3. Similar 14C-kinetic data were found for 14C-elution with and without chewing simulation with the Fatigue-machine and with MUC-3. During the first 26 h after the beginning of the experiments a linear 14C-elution kinetic was observed, followed by a second linear 14C-elution kinetic with a lower slope up to 86 h in both apparatus. It could be shown that chewing simulation has no significant (p<0.05) effect on the release of 14C-TEGDMA (and/or 14C-degradation products) from polymerized composites.

Localization and functional characterization of the human NKCC2 isoforms.

AIM: Salt reabsorption across the apical membrane of cells in the thick ascending limb (TAL) of Henle is primarily mediated by the bumetanide-sensitive Na(+)/K(+)/2Cl(-) cotransporter NKCC2. Three full-length splice variants of NKCC2 (NKCC2B, NKCC2A and NKCC2F) have been described. The NKCC2 isoforms have specific localizations and transport characteristics, as assessed for rabbit, rat and mouse. In the present study, we aimed to address the localization and transport characteristics of the human NKCC2 isoforms. METHODS: RT-PCR, in situ hybridization and uptake studies in Xenopus oocytes were performed to characterize human NKCC2 isoforms. RESULTS: All three classical NKCC2 isoforms were detected in the human kidney; in addition, we found splice variants with tandem duplicates of the variable exon 4. Contrary to rodents, in which NKCC2F is the most abundant NKCC2 isoform, NKCC2A was the dominant isoform in humans; similarly, isoform-specific in situ hybridization showed high expression levels of human NKCC2A along the TAL. Compared to NKCC2B and NKCC2F, human NKCC2A had the lowest Cl(-) affinity as determined by (86)Rb(+) uptake studies in oocytes. All NKCC2 isoforms were more efficiently inhibited by bumetanide than by furosemide. A sequence analysis of the amino acids encoded by exon 4 variants revealed high similarities between human and rodent NKCC2 isoforms, suggesting that differences in ion transport characteristics between species may be related to sequence variations outside the highly conserved sequence encoded by exon 4. CONCLUSION: The human NKCC2 is an example of how differential splicing forms the basis for a diversification of transporter protein function.

Rapamycin inhibits oncogenic intestinal ion channels and neoplasia in APC(Min/+) mice.

The adenomatous polyposis coli (APC) gene is mutated in familial adenomatous polyposis. Mice with a heterozygous APC(Min) mutation develop multiple intestinal neoplasia (Min) leading to premature death. Early in colorectal carcinogenesis, APC(Min/+) mice show enhanced Akt-mammalian target of rapamycin (mTOR) signaling, which is paralleled by upregulation of oncogenic K(+) channels. In this study, we tested the effect of mTOR inhibition with rapamycin on tumor formation in APC(Min/+) mice and evaluated ion channel regulation. We found that continuous long-term rapamycin treatment of APC(Min/+) mice dramatically inhibits intestinal neoplasia. Moreover, although untreated APC(Min/+) mice lose weight, experience intestinal bleeding and succumb to multiple neoplasia by 22.3+/-1.4 weeks of age, mice treated with rapamycin maintain stable weight and survive long term (39.6+/-3.4 weeks), with more than 30% surviving >1 year. Impressively, abnormalities in colonic electrolyte transport typical for APC(Min/+) mice are abolished, along with the suppression of epithelial Na(+) channel (ENaC) and oncogenic K(+) ion channels BK, Elk1 and Erg1, both functionally and at mRNA levels. These results show that continuous prophylaxis by rapamycin markedly inhibits the development of APC mutation-related polyposis, and suggest a novel contributing mechanism of action through the blockade of intestinal oncogenic ion channels.

Distribution and excretion of BisGMA in guinea pigs.

Bisphenol-A-glycidyldimethacrylate (BisGMA) is used in many resin-based dental materials. It was shown in vitro that BisGMA was released into the adjacent biophase from such materials during the first days after placement. In this study, the uptake, distribution, and excretion of [(14)C]BisGMA applied via gastric and intravenous administration (at dose levels well above those encountered in dental care) were examined in vivo in guinea pigs to test the hypothesis that BisGMA reaches cytotoxic levels in mammalian tissues. [(14)C]BisGMA was taken up rapidly from the stomach and intestine after gastric administration and was widely distributed in the body following administration by each route. Most [(14)C] was excreted within one day as (14)CO(2). The peak equivalent BisGMA levels in guinea pig tissues examined were at least 1000-fold less than known toxic levels. The peak urine level in guinea pigs that received well in excess of the body-weight-adjusted dose expected in humans was also below known toxic levels. The study therefore did not support the hypothesis.

Ca2+-induced Cl- efflux at rat distal colonic epithelium.

With the aid of the halide-sensitive dye 6-methoxy-N-ethylquinolinium iodide (MEQ), changes in intracellular Cl(-) concentration were measured to characterize the role of Ca(2+)-dependent Cl(-) channels at the rat distal colon. In order to avoid indirect effects of secretagogues mediated by changes in the driving force for Cl(-) exit (i.e., mediated by opening of Ca(2+)-dependent K(+) channels), all experiments were performed under depolarized conditions, i.e., in the presence of high extracellular K(+) concentrations. The Ca(2+)-dependent secretagogue carbachol induced a stilbene-sensitive Cl(-) efflux, which was mimicked by the Ca(2+) ionophore ionomycin. Surprisingly, the activation of Ca(2+)-dependent Cl(-) efflux was resistant against blockers of classical Ca(2+) signaling pathways such as phospholipase C, protein kinase C and calmodulin. Hence, alternative pathways must be involved in the signaling cascade. One possible signaling molecule seems to be nitric oxide (NO) as the NO donor sodium nitroprusside could induce Cl(- )efflux. Vice versa, the NO synthase inhibitor N-omega-monomethyl-arginine (L: -NMMA) reduced the carbachol-induced Cl(- )efflux. This indicates that NO may be involved in part of the signaling cascade. In order to test the ability of the epithelium to produce NO, the expression of different isoforms of NO synthase was verified by immunohistochemistry. In addition, the cytoskeleton seems to play a role in the activation of Ca(2+)-dependent Cl(-) channels. Inhibitors of microtubule association such as nocodazole and colchicine as well as jasplakinolide, a drug that enhances actin polymerization, inhibited the carbachol-induced Cl(-) efflux. Consequently, the activation of apical Cl(-) channels by muscarinic receptor stimulation differs in signal transduction from the classical phospholipase C/protein kinase C way.

KCNMA1 gene amplification promotes tumor cell proliferation in human prostate cancer.

Molecular mechanisms of prostate cancer progression are poorly understood. Here, we studied gene amplification of the large conductance calcium-activated potassium channel alpha subunit (KCNMA1), which is located at the chromosomal region 10q22. Fluorescence in situ hybridization (FISH) revealed KCNMA1 amplification in 16% of 119 late-stage human prostate cancers and in the hormone-insensitive prostate cancer cell line PC-3. In contrast, KCNMA1 amplification was absent in 33 benign controls, 32 precursor lesions and in 105 clinically organ-confined prostate cancers. Amplification was associated with mRNA and protein overexpression as well as increased density of BK channel protein and beta-estradiol-insensitive BK currents in PC-3 cells as compared to non-amplified control cell lines. Specific blockade of BK channels by iberiotoxin or RNA(i) significantly inhibited K(+) currents and growth of PC-3 cells. The data demonstrate that 10q22 amplification drives KCNMA1 expression and cell proliferation. Thus, KCNMA1 qualifies as a promising diagnostic and therapeutic target in patients with prostate cancer.

Antagonist enamel wears more than ceramic inlays.

Wear phenomena of ceramic inlays are not fully understood. The aim of the present study was to evaluate ceramic wear, antagonist enamel wear, and luting cement wear over 8 years. The two-fold null hypothesis was that there would be (1) no difference in wear behavior between ceramic and enamel, and (2) no influence of filler content of luting composites on composite wear. From 96 restorations, 36 Class II inlays from 16 participants were selected. For inlays with opposing enamel cusps (n=17), replicas of inlays and enamel were scanned with a 3-D laser scanner. Luting gaps of inlays (n=36) were analyzed with a profilometer, including 3-D data analysis. Ceramic and enamel wear increased between 4 and 8 years, with significantly higher values for enamel after 6 years (p<0.05). Luting gap wear increased continuously up to 8 years (p<0.05), with no influence of luting composites (p>0.05) and location of teeth (p>0.05).

Purinergic inhibition of the epithelial Na+ transport via hydrolysis of PIP2.

Stimulation of purinergic receptors inhibits amiloride-sensitive Na+ transport in epithelial tissues by an unknown mechanism. Because previous studies excluded the role of intracellular Ca2+ or protein kinase C, we examined whether purinergic regulation of Na+ absorption occurs via hydrolysis of phospholipid such as phosphatidylinositol-bisphosphates (PIP2). Inhibition of amiloride-sensitive short-circuit currents (Isc-Amil) by adenine 5'-triphosphate (ATP) in native tracheal epithelia and M1 collecting duct cells was suppressed by binding neomycin to PIP2, and recovery from ATP inhibition was abolished by blocking phosphatidylinositol-4-kinase or diacylglycerol kinase. Stimulation by ATP depleted PIP2 from apical membranes, and PIP2 co-immunoprecipitated the beta subunit of ENaC. ENaC was inhibited by ATP stimulation of P2Y2 receptors in Xenopus oocytes. Mutations in the PIP2 binding domain of betaENaC but not gammaENaC reduced ENaC currents without affecting surface expression. Collectively, these data supply evidence for a novel and physiologically relevant regulation of ENaC in epithelial tissues. Although surface expression is controlled by its C terminus, N-terminal binding of betaENaC to PIP2 determines channel activity.

Control of epithelial ion transport by Cl- and PDZ proteins.

Inhibition of epithelial Na+ channels (ENaC) by the cystic fibrosis transmembrane conductance regulator (CFTR) has been demonstrated previously. Recent studies suggested a role of cytosolic Cl- for the interaction of CFTR with ENaC, when studied in Xenopus oocytes. In the present study we demonstrate that the Na+ / H+ -exchanger regulator factor (NHERF) controls expression of CFTR in mouse collecting duct cells. Inhibition of NHERF largely attenuates CFTR expression, which is paralleled by enhanced Ca(2+) -dependent Cl- secretion and augmented Na+ absorption by the ENaC. It is further demonstrated that epithelial Na+ absorption and ENaC are inhibited by cytosolic Cl- and that stimulation by secretagogues enhances the intracellular Cl- concentration. Thus, the data provide a clue to the question, how epithelial cells can operate as both absorptive and secretory units: Increase in intracellular Cl- during activation of secretion will inhibit ENaC and switch epithelial transport from salt absorption to Cl- secretion.

Polymerization contraction stress in light-cured compomer restorative materials.

OBJECTIVE: The magnitude and kinetics of polymerization contraction stress build-up may be potential predictors of bond failure of adhesive restorations. The present study determined these properties of seven commercial compomers (Dyract, Dyract AP, F2000 Rasant, Hytac, Compoglass F, Luxat, Glasiosite). METHODS: Polymerization shrinkage was generated by 40 s light curing the test materials (800 mW/cm2). The contraction force induced was recorded for 300 s at room temperature (23-24 degrees C) by means of a Stress-Strain-Analyzer (C factor=0.33). Maximum contraction stress (MPa), coefficient of near linear fit of contraction force/time (gradient) and relative force rate (%/s) of each material were compared with that of two hybrid composites (Tetric Ceram, Prodigy). The statistical analysis was conducted by ANOVA (alpha=0.05) and post hoc Tukey's test. RESULTS: No statistically significant differences in the maximum stress between Glasiosite (2.27+/-0.06 MPa), Hytac (2.31+/-0.07 MPa) and Tetric Ceram (2.21+/-0.11 MPa), and between Compoglass F (2.60+/-0.18 MPa) and Prodigy (2.70+/-0.06 MPa) were found. The contraction stress of F2000 Rasant (3.41+/-0.09 MPa) and Luxat (3.33+/-0.08 MPa) were significantly highest, whilst Dyract exhibited the significantly lowest shrinkage stress (1.27+/-0.08 MPa) among the tested materials. SIGNIFICANCE: High contraction stress, early start of stress build-up and rapid contraction force development may lead to failure of bond to tooth structure. This study suggested that the contraction stress and kinetic behavior of compomers are generally similar to those of hybrid composites in a dry condition. Dyract might be superior in maintaining the bond with cavity walls compared to conventional hybrid composites in view of its low shrinkage stress.