5.5-year-survival of CAD/CAM resin-based composite restorations in severe tooth wear patients.

OBJECTIVES: Aim of this prospective study was to assess full mouth rehabilitation of severe tooth wear patients using minimally invasive CAD/CAM resin-based composite (RBC) restorations and direct veneers by evaluating restoration survival up to 5.5-years. METHODS: Twenty-two patients with generalized severe tooth wear with functional and/or esthetic problems were included. Following minimally invasive preparation, CAD/CAM RBC restorations (LAVA Ultimate,3M) were adhesively luted, direct RBC veneers (Filtek Supreme XTE, 3M) were applied in the aesthetic region. Patients were recalled after 1m,1y,3y,5y and seen in between recalls by their general dentists or at the clinical study center if complaints occurred. Failures were categorized as F1 (severe deficiencies requiring replacement/extraction), F2 (localized deficiencies requiring re-cementation/repair) and F3 (small chippings requiring refurbishment/monitoring). Survival of indirect restorations was evaluated using lifetables and Kaplan-Meier-graphs, distinguishing between failure categories and tooth type (front teeth=FT, premolars=PM, molars=M). F1 + F2 and F1 + F2 + F3 failures were analyzed using Cox regression on the variables tooth type/ location, age, gender and VDO increase (p < 0.05). RESULTS: 568 indirect restorations and 200 direct veneers in 21 patients evaluated for up to 5.5-years. For indirect restorations, 96 failures were recorded (F1:6;F2:41;F3:49) and annual failure rates were 0.29%(FT), 1.56%(PM), 2.93%(M) for F1 +F2 and 0.53%(FT), 2.42%(PM), 6.11%(M) for F1 + F2 + F3. Reasons for failure were chipping fracture (48), adhesive fracture (32), complete debonding (7), caries (4), endodontic treatment (1) and reasons unknown (documentation general dentists, 4). Molar tooth type had a statistically significantly increased probability of failure compared with front teeth and premolars for F1 + F2 + F3 (p < 0.006). Direct veneer restorations showed 18 failures (F1:2;F2:9;F3:7). SIGNIFICANCE: Minimally invasive CAD/CAM RBC restorations combined with direct RBC veneers showed an acceptable clinical mid-term survival for restorative rehabilitation of severely worn dentitions.

A Prospective Comparative Analysis Regarding the Assessment of Oral Mucosal Disease Using a Validated Questionnaire to Improve the Teaching of Dental Students.

Structured examination and treatment are essential in medicine. For dental students, a structured approach to the assessment of oral mucosal lesions is missing thus far. To validate an approach, a structured questionnaire was compared with the habitually used free description of oral lesions (white lesions, ulcers, hyperplasia). Thirty-three dental students were divided into two groups (Group 1 (n = 17) used the free description; Group 2 (n = 16) used a guided questionnaire) to characterize mucosal lesions in patients and make a tentative diagnosis. Although no difference was found between the groups regarding the suspected diagnosis or the histopathological findings, there was a significant advantage of the structured questionnaire in all aspects of the description compared to the free description (p = 0.000018). Thus, a structured description is an important aspect in the evaluation of oral mucosal changes, and a guided questionnaire should be implemented in the study of dentistry.

Fourteen years clinical evaluation of leucite-reinforced ceramic inlays luted using two different adhesion strategies.

OBJECTIVES: Aim of the present prospective study was to clinically evaluate the long-term performance of two different luting-materials for leucite-reinforced glass-ceramic inlays/onlays after 14 years. METHODS: A total of 83 IPS-Empress-inlays/onlays were placed in 30 patients. Restorations were luted according to two different strategies: 43 restorations were fixed with a self-adhesive resin-cement (RelyXUnicem, RX), 40 restorations were inserted with VariolinkII-low (SV) after pretreatment with an etch-and-rinse multi-step adhesive. Recalls were performed after two weeks (n=83), two years (n= 82), four years (n=74) and 14 years (n=54). Two independent calibrated examiners evaluated all restorations using modified USPHS-criteria. Statistical analysis was performed using pairwise Mann-Whitney-U-test and Friedman-test (p < 0.05). RESULTS: After 14 years, 54 restorations in 22 patients were evaluated (eight patients equalling 29 inlays not available). Ten restorations had to be replaced (failure rate 12%); four (SV-group) showed bulk fractures and two (RX-group) exhibited marginal fractures at the 14-year recall. Overall, the SV-group revealed significantly better results regarding discoloration of the luting gap (p<0.05) compared to the RX-group. No statistically significant differences were computed between SV and RX for the remaining criteria at the respective recalls (p>0.05). However, statistically significant deteriorations were detected for both luting procedures over 14 years regarding "colour match", "marginal integrity" and "tooth integrity" (p<0.05). CONCLUSIONS: The self-adhesive resin-cement RelyXUnicem showed similar clinical performance to a conventional multi-step luting-procedure after 14 years for most of the test parameters with a slightly inferior performance of RelyXUnicem regarding discoloration of the luting gap. CLINICAL SIGNIFICANCE: The current study presents unique in-vivo long-term data on two adhesion-strategies for indirect ceramic single-tooth restorations. Differences in performance of the two luting methods after being challenged for 14 years in the oral environment are highlighted. However, the overarching survival rate justifies the recommendation of both methods for clinical routine.

Evaluation of wear behavior of dental restorative materials against zirconia in vitro.

OBJECTIVES: To evaluate two-body wear (2BW) and three-body wear (3BW) of different CAD/CAM and direct restorative materials against zirconia using a dual-axis chewing simulator and an ACTA wear machine. METHODS: 3 CAD-CAM resin-based composite or polymer infiltrated ceramic network blocs, 1 lithium disilicate CAD-CAM ceramic (LS2), 3 direct resin composites, amalgam and bovine enamel were tested. For 2BW, 8 flat specimens per material were produced, grinded, polished, stored wet (37 °C, 28d) and tested (49 N, 37 °C, 1,200,000 cycles) against zirconia. For 3BW, specimens (n = 10) were stored accordingly, and tested against a zirconia antagonist wheel (3Y-TZP, d = 20 mm, h = 6 mm; 200,000 cycles, F = 15 N, f = 1 Hz, 15% slip) in millet seed suspension. Wear resistance was analysed in a 3D optical non-contact profilometer, measuring vertical wear depth and volume loss for 2BW and mean wear depth and roughness (Ra) for 3BW. Vickers hardness (15 s, HV2) was measured. Statistical analysis was performed using non-parametric tests (Mann-Whitney-U test, p < 0.05). RESULTS: 2BW and 3BW have a different impact on material surfaces. Similar wear resistance was observed for direct and indirect resin based materials with analogous filler configurations in both methods. Bovine enamel exhibited the best wear resistance in 2BW, but the least wear resistance in 3BW against zirconia. Regarding 2BW, a direct/indirect composite material pair of the same manufacturer showed the significantly highest mean volume losses (2.72/2.85 mm³), followed by LS2 (1.41 mm³). LS2 presented the best wear resistance in 3BW (mean wear depth 2.85 µm), combined with the highest mean Vickers hardness (598 MPa). No linear correlation was found between Vickers hardness and both wear testing procedures. The zirconia antagonists showed no recordable signs of wear. SIGNIFICANCE: Dental restorative materials behave differently in 2BW and 3BW laboratory testing. Vickers hardness testing alone cannot hold for a correlation with wear behavior of materials. Micromorphological investigation of material composition can reveal insights in wear mechanisms related to variations in filler technologies.

Oxygen concentration of gut luminal contents varies post-prandially in growing pigs.

The oxygen (O2 ) concentration of gastrointestinal tract (GIT) contents decreases distally, but little is known about how O2 concentrations are influenced by ingestion of a meal. The O2 concentration in luminal contents at different GIT locations (stomach [cardia and pylorus], proximal, mid- and distal small intestine and caecum) and how these concentrations changed post-prandially were determined. Fifty entire male pigs (22 kg bodyweight at the start of study) were fed semi-synthetic diets containing casein, α-lactalbumin, whey protein isolate or zein as the sole source of protein for 8 days. A further group of pigs received the casein diet for six days and a semi-synthetic protein-free diet for a further 2 days. On day 8, pigs (n = 2 per diet and time point) were euthanized post-prandially (0, 1, 2, 4 and 6 h), and the stomach, small intestine and caecum were isolated and O2 determined in the GIT contents. Observations at each time point were averaged across the diets (n = 10). The mean O2 concentration was markedly higher (p ≤ 0.05) in the stomach compared with the rest of the GIT. The O2 concentration was similar in the small intestinal regions (p > 0.05; 1.0%-1.1%) and the caecum (0.9%), apart for the proximal small intestine which had a 24% higher (p ≤ 0.05) O2 concentration than the caecum. The mean O2 concentration in the GIT varied post-prandially (p ≤ 0.05). The O2 concentration in the cardia decreased 1.8%/h over the first two hours post-feeding and thereafter increased 0.3%/h (p ≤ 0.05). In the caecum, the O2 concentration was constant during the first 4 h and thereafter increased slightly (p ≤ 0.05). The flow of food through the GIT influenced both the concentration and amount of O2 in GIT luminal contents.

Usability of abattoir-acquired pig eyes for refractive excimer laser research.

The purpose of this study was to elucidate, under which conditions abattoir-acquired pig eyes are suitable for refractive excimer laser experiments. Porcine eyes from tunnel-scalded (n = 5) and tank-scalded (n = 10) pigs were compared to unscalded eyes (n = 5) and to eyes scalded in the laboratory (n = 5). The corneal epithelium was removed before an excimer laser was used to perform a - 8.0 D photoablation. Corneal thickness was measured by optical coherence topography before and after photoablation. The ablation depth was determined with a contour measuring station, the morphology of the ablated areas was characterized by scanning electron microscopy and white-light profilometry. The scalded eyes showed an increase in corneal swelling which gained statistical significance in tank-scalded eyes showing a wedge-shaped opaque stromal lesion in the nasal corneal quadrant. A measurable deterioration of photoablation was only found in tank-scalded eyes that exhibited the opaque lesion. Ablated area morphology was smooth and regular in the unscalded and tunnel-scalded eyes. The tank-scalded eyes showed conspicuous wrinkles. While unscalded eyes should always be preferred for excimer laser laboratory experiments, the data suggest that the use of tunnel-scalded eyes may also be acceptable and should be chosen over tank-scalded eyes.

Fracture toughness of dental incremental composite-composite interfaces at elevated temperatures.

The aim of the present laboratory study was to mechanically characterize the interface between two dental resin-based composite (RBC) increments, and to investigate if elevated temperatures have an influence on the quality of the interface mimicking clinical filling procedure. Four RBCs (CLEARFIL MAJESTY™ Posterior, Kuraray (CMP)/Filtek™ Supreme XTE, 3M (FSX)/Grandio®SO, VOCO (GSO)/VisCalor® bulk, VOCO (VCB)) were tested with a fracture toughness test using Chevron notched beams (KI,CNB) at 23, 37 and 54 °C. KI,CNB specimens (3 × 4x25mm) with a V-shaped notch at the incremental interface were loaded until failure in a 4-point bending set-up. Failure modes were characterized using light microscopy, microstructural interface was analyzed using SEM. Statistical analysis was performed using Kolmogorov-Smirnoff test, two-way ANOVA and Tukey Post-Hoc test (p = 0.05). Mean KI,CNB ranged between 0.73 ±0.14 MPam0.5 (VCB, 23 °C) and 1.11 ± 0.11 MPam0.5 (FSX, 23 °C). The tested conventional highly filled RBCs presented fracture toughness at the incremental interface comparable to the cohesive strength of the bulk materials. VCB showed reduced interfacial fracture toughness at 23 and 37 °C, but performed well at elevated temperature of 54 °C.

Viscosity and stickiness of dental resin composites at elevated temperatures.

OBJECTIVE: To investigate the influence of pre-heating different classes of dental resin composites on viscosity and stickiness at five different temperatures. METHODS: Six flowable, five conventional packable, and one thermo-viscous bulk-fill resin composites were heated up to 54°C in a plate-plate rheometer to determine their complex viscosity. Normal force measurements were carried out for the six packable materials to determine the unplugging force and unplugging work (stickiness) over the same temperature range. Data were analyzed using Kolmogorov-Smirnoff test, one-way ANOVA and Tukey Post Hoc test with α=0.05 as level of significance. RESULTS: At 23°C packable composites showed viscosity between 6.75 and 19.14kPas, while flowable composites presented significantly lower viscosities between 1.31 and 2.20kPas. Pre-heating led to a drop of 30-82% in the viscosity of packable materials. The thermo-viscous material dropped to the level of flowables at 45 and 54°C thus behaving as a packable composite at room temperature with flowable-like viscosity at higher temperatures. No statistically significant differences for viscosity were observed among flowable composites at any temperature. The unplugging force decreased for packable composites, while their unplugging work generally increased at elevated temperature. At 23°C unplugging force was measured between 7.50 and 19.18N, while pre-heating up to 54°C led to values between 2.9 and 6.2N. Regarding unplugging work at 23°C the calculated values were between 3.0 and 8.9×10-3J and at 54°C between 8.8 and 13.0×10-3J. SIGNIFICANCE: Pre-heating significantly reduced viscosity of highly viscous resin composites, while no influence was shown for flowable composites. In general stickiness, measured as unplugging work, increased at elevated temperatures. The thermo-viscous material showed low viscosity comparable to flowable composites at 45 and 54°C, yet its stickiness did not increase significantly compared to the values at 23°C.

New Approaches in Bonding to Glass-Ceramic: Self-Etch Glass-Ceramic Primer and Universal Adhesives.

PURPOSE: To investigate the tensile bond strength of silane-containing universal adhesives and self-etch glass-ceramic primer to lithium disilicate glass ceramics (LS2). MATERIALS AND METHODS: 960 rectangular LS2 bars (7 mm x 3 mm x 9 mm, IPS e.max CAD, Ivoclar Vivadent) were manufactured and divided into 4 groups (n = 240). Group 1 was etched with ~5% hydrofluoric acid (HF) for 20 s (VITA Ceramics Etch, Vita Zahnfabrik), group 2 was etched with ~5% HF for 20 s and silanized (ESPE Sil, 3M Oral Care), group 3 was pre-treated with a self-etching glass-ceramic primer (Monobond Etch & Prime, Ivoclar Vivadent, and group 4 received no pre-treatment. Three universal adhesives (iBOND Universal, Heraeus Kulzer; Scotchbond Universal Adhesive, 3M Oral Care; Futurabond U, Voco) were applied to the differently pre-treated surfaces, with Heliobond (Ivoclar Vivadent) serving as control. The bars from each group were paired and luted perpendicularly, forming a square bonded area of 9 mm2, using Variolink II (Ivoclar Vivadent) with a constant pressure of 10 N, followed by light curing (40 s at 800 mW/cm2, Elipar Trilight, 3M Oral Care). The resulting specimens were stored for 24 h at 37°C in distilled water. Half of the specimens of each group were submitted to tensile bond strength testing, the other half were thermocycled ([TC] 5000 cycles, 5°C/55°C, 30-s dwell time) before testing. Data were analyzed using three-way ANOVA (α = 0.05). RESULTS: Group 2 (HF etched and silanized) and group 3 (self-etching glass-ceramic primer) reached significantly higher mean bond strengths than did groups 1 (only HF etched) and 4 (no pre-treatment). CONCLUSION: Additional silanization of HF-etched LS2 statistically signficantly improved the tensile bond strength of the silane-containing universal adhesive (Scotchbond Universal). The self-etching glass-ceramic primer Monobond Etch & Prime achieved mean bond strengths that did not differ significantly from HF-etched and silanized specimens.

Dual Targeting of Y-Box Binding Protein-1 and Akt Inhibits Proliferation and Enhances the Chemosensitivity of Colorectal Cancer Cells.

KRAS-mutated colorectal cancers (CRCs) are resistant to cetuximab treatment. The multifunctional Y-box binding protein 1 (YB-1) is overexpressed in CRC and is associated with chemoresistance. In this study, the effects of oncogenic mutated KRAS(G12V) and KRAS(G13D) on YB-1 phosphorylation were investigated in CRC cells. The effects of the inhibition of p90 ribosomal S6 kinase (RSK) on YB-1 phosphorylation, cell proliferation and survival were tested with and without treatment with 5-fluorouracil using pharmacological inhibitors and siRNA. YB-1 phosphorylation status and subcellular distribution in CRC patient tissues were determined by immunofluorescence staining and confocal microscopy. Endogenous expression of mutated KRAS(G13D) and conditional expression of KRAS(G12V) significantly stimulated YB-1 phosphorylation via RSK and were associated with cetuximab resistance. Inhibition of YB-1 by targeting RSK stimulated the Akt signaling pathway, and this stimulation occurred independently of KRAS mutational status. Akt activation interfered with the antiproliferative effect of the RSK inhibitor. Consequently, dual targeting of RSK and Akt efficiently inhibited cell proliferation in KRAS(G13D)-mutated HCT116 and KRAS wild-type SW48 cells. Treatment with 5-fluorouracil (5-FU) significantly enhanced YB-1 phosphorylation in KRAS(G13D)-mutated HCT116 cells but not in KRAS wild-type SW48 cells. Dual targeting of Akt and RSK sensitized HCT116 cells to 5-FU by stimulating 5-FU-induced apoptosis and inhibiting repair of 5-FU-induced DNA damage. YB-1 was highly phosphorylated in CRC patient tumor tissues and was mainly localized in the nucleus. Together, dual targeting of RSK and Akt may be an alternative molecular targeting approach to cetuximab for treating CRC in which YB-1 is highly phosphorylated.

Stress-Induced Phosphorylation of Nuclear YB-1 Depends on Nuclear Trafficking of p90 Ribosomal S6 Kinase.

Ionizing radiation (IR) and epidermal growth factor (EGF) stimulate Y-box binding protein-1 (YB-1) phosphorylation at Ser-102 in KRAS wild-type (KRASwt) cells, whereas in KRAS mutated (KRASmut) cells, YB-1 is constitutively phosphorylated, independent of IR or EGF. YB-1 activity stimulates the repair of IR-induced DNA double-strand breaks (DSBs) in the nucleus. Thus far, the YB-1 nuclear translocation pattern after cell exposure to various cellular stressors is not clear. In the present study, we investigated the pattern of YB-1 phosphorylation and its possible translocation to the nucleus in KRASwt cells after exposure to IR, EGF treatment, and conditional expression of mutated KRAS(G12V). IR, EGF, and conditional KRAS(G12V) expression induced YB-1 phosphorylation in both the cytoplasmic and nuclear fractions of KRASwt cells. None of the stimuli induced YB-1 nuclear translocation, while p90 ribosomal s6 kinase (RSK) translocation was enhanced in KRASwt cells after any of the stimuli. EGF-induced RSK translocation to the nucleus and nuclear YB-1 phosphorylation were completely blocked by the EGF receptor kinase inhibitor erlotinib. Likewise, RSK inhibition blocked RSK nuclear translocation and nuclear YB-1 phosphorylation after irradiation and KRAS(G12V) overexpression. In summary, acute stimulation of YB-1 phosphorylation does not lead to YB-1 translocation from the cytoplasm to the nucleus. Rather, irradiation, EGF treatment, or KRAS(G12V) overexpression induces RSK activation, leading to its translocation to the nucleus, where it activates already-existing nuclear YB-1. Our novel finding illuminates the signaling pathways involved in nuclear YB-1 phosphorylation and provides a rationale for designing appropriate targeting strategies to block YB-1 in oncology as well as in radiation oncology.

Live Faecalibacterium prausnitzii induces greater TLR2 and TLR2/6 activation than the dead bacterium in an apical anaerobic co-culture system.

Inappropriate activation of intestinal innate immune receptors, such as toll-like receptors (TLRs), by pathogenic bacteria is linked to chronic inflammation. In contrast, a "tonic" level of TLR activation by commensal bacteria is required for intestinal homeostasis. A technical challenge when studying this activation in vitro is the co-culturing of oxygen-requiring mammalian cells with obligate anaerobic commensal bacteria. To overcome this, we used a novel apical anaerobic co-culture system to successfully adapt a TLR activation assay to be conducted in conditions optimised for both cell types. Live Faecalibacterium prausnitzii, an abundant obligate anaerobe of the colonic microbiota, induced higher TLR2 and TLR2/6 activation than the dead bacterium. This enhanced TLR induction by live F. prausnitzii, which until now has not previously been described, may contribute to maintenance of gastrointestinal homeostasis. This highlights the importance of using physiologically relevant co-culture systems to decipher the mechanisms of action of live obligate anaerobes.

Live Faecalibacterium prausnitzii Does Not Enhance Epithelial Barrier Integrity in an Apical Anaerobic Co-Culture Model of the Large Intestine.

Appropriate intestinal barrier maturation during infancy largely depends on colonization with commensal bacteria. Faecalibacterium prausnitzii is an abundant obligate anaerobe that colonizes during weaning and is thought to maintain colonic health throughout life. We previously showed that F. prausnitzii induced Toll-like receptor 2 (TLR2) activation, which is linked to enhanced tight junction formation. Therefore, we hypothesized that F. prausnitzii enhances barrier integrity, an important factor in appropriate intestinal barrier maturation. In order to test metabolically active bacteria, we used a novel apical anaerobic co-culture system that allows the survival of both obligate anaerobic bacteria and oxygen-requiring intestinal epithelial cells (Caco-2). The first aim was to optimize the culture medium to enable growth and active metabolism of F. prausnitzii while maintaining the viability and barrier integrity, as measured by trans-epithelial electrical resistance (TEER), of the Caco-2 cells. This was achieved by supplementing the apical cell culture medium with bacterial culture medium. The second aim was to test the effect of F. prausnitzii on TEER across Caco-2 cell layers. Live F. prausnitzii did not improve TEER, which indicates that its benefits are not via altering tight junction integrity. The optimization of the novel dual-environment co-culturing system performed in this research will enable the investigation of new probiotics originating from indigenous beneficial bacteria.

MicroRNAs and their role for T stage determination and lymph node metastasis in early colon carcinoma.

Worldwide, colon cancer is among the most common cancer entities. Understanding the molecular background is the key to enable accurate stage determination, which is crucial to assess optimal therapy options. The search for preoperative biomarkers is ongoing. In recent years, several studies have proposed a diagnostic and prognostic role for miRNAs in cancer. Aim of this study was to evaluate miRNA expression patterns correlating with tumor stage, especially lymph node metastasis, in primary colon carcinoma tissue. Screening was accomplished using GeneChip® miRNA v3.0 arrays (Thermo Fisher Scientific, Waltham, MA, USA) and validated via TaqMan® qPCR assays (Thermo Fisher Scientific, Waltham, MA, USA) to investigate miRNA expressions in 168 FFPE and 83 fresh frozen colon carcinoma samples. Regarding lymph node status, analyses displayed no significantly differential miRNA expression. Interestingly, divergent expression of miR-18a-5p, miR-20a-5p, miR-21-5p, miR-152-3p and miR-1973 was detected in stage pT1. Although miRNAs might not represent reliable biomarkers regarding lymph node metastasis status, they could support risk assessment in stage T1 tumors.

Human oral isolate Lactobacillus fermentum AGR1487 induces a pro-inflammatory response in germ-free rat colons.

Lactobacilli are thought to be beneficial for human health, with lactobacilli-associated infections being confined to immune-compromised individuals. However, Lactobacillus fermentum AGR1487 negatively affects barrier integrity in vitro so we hypothesized that it caused a pro-inflammatory response in the host. We compared germ-free rats inoculated with AGR1487 to those inoculated with another L. fermentum strain, AGR1485, which does not affect in vitro barrier integrity. We showed that rats inoculated with AGR1487 had more inflammatory cells in their colon, higher levels of inflammatory biomarkers, and increased colonic gene expression of pro-inflammatory pathways. In addition, our in vitro studies showed that AGR1487 had a greater capacity to activate TLR signaling and induce pro-inflammatory cytokines in immune cells. This study indicates the potential of strains of the same species to differentially elicit inflammatory responses in the host and highlights the importance of strain characterization in probiotic approaches to treat inflammatory disorders.

Bulk-fill resin composites: polymerization properties and extended light curing.

OBJECTIVES: The aim was to evaluate the polymerization properties of bulk-fill resin composites using two different light-curing protocols, in terms of degree of conversion (%DC), Vickers hardness (HV), polymerization volume shrinkage (PVS) and polymerization shrinkage stress (PSS) and compare them to conventional condensable and flowable resin composites. MATERIALS AND METHODS: Filtek BulkFill (FBF, 3MESPE, Germany), SDR (Dentsply, Germany), TetricEvoCeram BulkFill (TBF, Ivoclar-Vivadent, Liechtenstein), Venus BulkFill (VBF, Heraeus, Germany), X-traBase (XTB, Voco, Germany), FiltekZ250 (3MESPE) and Filtek Supreme XTE Flowable (FSF, 3MESPE) were investigated. Light-curing was performed for 30 s or according to manufacturers' instructions (1200 mW/cm2, Bluephase20i, Ivoclar-Vivadent). For %DC and HV, discs (n=5) of 2 or 4 mm in thickness were prepared and stored for 24h in distilled water at 37°C. %DC was determined by FTIR-ATR-spectroscopy. %DC and HV were measured at the top and bottom of the specimens. PVS was measured using Archimedes method (n=6). PSS measurements (n=10) were carried out using 5 mm diameter PMMA rods as bonding substrates with a specimen height of 1 mm in a universal testing machine. Data were analyzed using one- and two-way ANOVA (α=0.05). RESULTS: Except Z250 in the manufacturers' light-curing mode, all materials showed no significant inferior %DC at 4 mm thickness. When light cured for 30 s Z250 had no significant differences in %DC at 2 or 4 mm when compared to top. FBF, TBF, FSF and Z250 displayed significant reduced HV at 4 mm in both curing modes. Z250 and TBF showed the lowest PVS and FSF the highest PSS in both curing modes. SIGNIFICANCE: All investigated bulk-fill composites obtained sufficient polymerization properties at 4 mm depth. Enhanced curing time improved the investigated polymerization properties of bulk-fills and Z250.

Understanding how commensal obligate anaerobic bacteria regulate immune functions in the large intestine.

The human gastrointestinal tract is colonised by trillions of commensal bacteria, most of which are obligate anaerobes residing in the large intestine. Appropriate bacterial colonisation is generally known to be critical for human health. In particular, the development and function of the immune system depends on microbial colonisation, and a regulated cross-talk between commensal bacteria, intestinal epithelial cells and immune cells is required to maintain mucosal immune homeostasis. This homeostasis is disturbed in various inflammatory disorders, such as inflammatory bowel diseases. Several in vitro and in vivo studies indicate a role for Faecalibacterium prausnitzii, Bacteroides thetaiotaomicron, Bacteroides fragilis, Akkermansia muciniphila and segmented filamentous bacteria in maintaining intestinal immune homeostasis. These obligate anaerobes are abundant in the healthy intestine but reduced in several inflammatory diseases, suggesting an association with protective effects on human health. However, knowledge of the mechanisms underlying the effects of obligate anaerobic intestinal bacteria remains limited, in part due to the difficulty of co-culturing obligate anaerobes together with oxygen-requiring human epithelial cells. By using novel dual-environment co-culture models, it will be possible to investigate the effects of the unstudied majority of intestinal microorganisms on the human epithelia. This knowledge will provide opportunities for improving human health and reducing the risk of inflammatory diseases.

Protective effects of Lactobacilli, Bifidobacteria and Staphylococci on the infection of cultured HT29 cells with different enterohemorrhagic Escherichia coli serotypes are strain-specific.

In this study, we investigated the interaction of 19 benign strains of lactic acid bacteria (LAB), bifidobacteria and staphylococci with enterohemorrhagic Escherichia coli (EHEC) strains of different serotypes and virulence gene spectrum in a HT29 cell culture infection model. As markers of infection, the secretion of interleukin 8 (IL-8) and the activation of the transcription factor NF-κB by the infected cells were determined. With 12 of 19 tested strains, a weak reduction <30% of IL-8 secretion of HT29 cells after co-infection with EHEC O157:H7 strain EDL933 was observed. Six strains reduced the IL-8 secretion up to 60% and the strain B. adolescentis DSMZ 20086 decreased the IL-8 production about 73%. In further co-infection assays with EHEC strains of the serotypes O103:H2, O26:H⁻, 0157:H⁻ and O113:H21, different abilities of the LAB strains to influence the infection with the different EHEC strains were noted. Therefore, the protective anti-inflammatory effect is strain specific for LAB and also depends on the application of EHEC strains with different sero- and virulence types. The differences in efficacy of protective bacteria against certain EHEC strains were unexpected and have not been shown so far. Furthermore, we could show that the inhibitory effects were not attributed to lower adhesion abilities of EHEC to the production of organic acids by the benign bacteria. In addition, viable bacteria are needed to inhibit the IL-8 secretion. Moreover, the NF-κB activation was reduced significantly by all tested LAB strains in co-infection trials, but was not strain-specific. The model described here is useful to screen for basic effects of protective bacteria that are able to counteract EHEC-mediated effects on human cells, and to study the molecular interaction between bacteria as well as between bacteria and human cultured cells.