Selective inhibition of V600E-mutant BRAF gene induces apoptosis in thyroid carcinoma cell lines.

PURPOSE: Papillary thyroid cancer (PTC) has a high incidence of BRAFV600E mutation. The purpose of this study was to evaluate the potential relationship between thyroiditis and BRAFV600E mutation status in patients with PTC. We investigated how a selective inhibitor of BRAFV600E PLX4032 affects the proliferation and inflammatory cytokine levels of thyroid cancer. METHODS: Two thyroid cancer cell lines TPC1 and 8505C were treated with PLX4032, an analysis was done on cell growth, cell cycle, the degree of apoptosis, and levels of inflammatory cytokines. To identify the functional links of BRAF, we used the STRING database. RESULTS: Docking results illustrated PLX4032 blocked the kinase activity by exclusively binding on the serine/threonine kinase domain. STRING results indicated BRAF is functionally linked to mitogen-activated protein kinase. Both cell lines showed a dose-dependent reduction in growth rate but had a different half maximal inhibitory concentration value for PLX4032. The reaction to PLX4032 was more sensitive in the 8505C cells than in the TPC1 cells. PLX4032 induced a G2/M phase arrest in the TPC1 cells and G0/G1 in the 8505C cells. PLX4032 induced apoptosis only in the 8505C cells. With PLX4032, the TPC1 cells showed decreased levels of vascular endothelial growth factor, granulocyte-macrophage colony-stimulating factor, chemokine (C-C motif) ligand 2/monocyte chemoattractant protein 1, whereas the 8505C cells showed significantly decreased levels of IL-8, serpin E1/plasminogen activator inhibitor-1, and matrix metalloproteinase (MMP)-3. CONCLUSION: PLX4032 was cytotoxic in both TPC1 and 8505C cells and induced apoptosis. In the 8505C cells, inflammatory cytokines such as IL-8 and MMP-3 were down-regulated. These findings suggest the possibility that the BRAFV600E mutation needs to target inflammatory signaling pathways in the treatment of thyroid cancer.

Prognostic Significance of Prostaglandin-Endoperoxide Synthase-2 Expressions in Human Breast Carcinoma: A Multiomic Approach.

Prostaglandin-endoperoxide synthase-2 (PTGS2) plays a pivotal role in inflammation and carcinogenesis in human breast cancer. Our aim of the study is to find the prognostic value of PTGS2 in breast cancer. We conducted a multiomic analysis to determine whether PTGS2 functions as a prognostic biomarker in human breast cancer. We explored PTGS2 mRNA expressions using different public bioinformatics portals. Oncomine, Serial Analysis of Gene Expression (SAGE), GEPIA, ULCAN, PrognoScan database, Kaplan-Meier Plotter, bc-GenExMiner, USC XENA, and Cytoscape/STRING DB were used to identify the prognostic roles of PTGS2 in breast cancer. Based on the clinicopathological analysis, decreased PTGS2 expressions correlated positively with older age, lymph node status, the human epidermal growth factor receptor 2 (HER2) status (P < .0001), estrogen receptor (ER+) expression (P < .0001) Luminal A (P < .0001), and Luminal B (P < .0001). Interestingly, progesterone receptor (PR) (P < .0001) negative showed a high expression of PTGS2. Prostaglandin-endoperoxide synthase-2 was downregulated in breast cancer tissues than in normal tissues. In the PrognoScan database and, Kaplan-Meier Scanner, downregulated expressions of PTGS2 associated with poor overall survival (OS), relapse-free survival (RFS), and distant metastasis-free survival. The methylation levels were significantly higher in the Luminal B subtype. Through oncomine coexpressed gene analysis, we found a positive correlation between PTGS2 and interleukin-6 (IL-6) expression in breast cancer tissues. These results indicate that downregulated expressions of PTGS2 can be used as a promising prognostic biomarker and Luminal B hyper methylation may play an important role in the development of breast cancers. However, to clarify our results, extensive study is required.

High-resolution dental magnetic resonance imaging for planning palatal graft surgery-a clinical pilot study.

AIM: To evaluate whether high-resolution, non-contrast-enhanced dental magnetic resonance imaging (MRI) can be used for accurate determination of palatal masticatory mucosa thickness (PMMT) and to locate the greater palatal artery (GPA). MATERIALS AND METHODS: In five volunteers (four males, one female; mean age 30.2 ± 0.4 years), two independent raters measured PMMT by use of dental MRI in 180 positions. For comparison, clinical bone sounding was performed. The GPA was identified in time-of-flight (TOF) angiography and MSVAT-SPACE-prototype sequence. Intra- and inter-observer agreement for MRI measurements, agreement between MRI and bone sounding were analysed by intra-class correlation coefficient (ICC) and Cohen's kappa (κ). RESULTS: Reliability of dental MRI measurements was high (intra-observer-ICC 0.962; inter-observer ICC 0.959). Agreement of MRI measurements with bone sounding was moderate (ICC 0.744), and the GPA could be identified in 60% of measurement points using the TOF-angiography alone and in 85% with additional information of the MSVAT-SPACE. Good intra-observer agreement was observed for GPA identification (κ: 0.778). CONCLUSION: Palatal masticatory mucosa thickness measured by high-resolution, non-contrast enhanced dental MRI is comparable with that obtained by bone sounding. Dental MRI enables reliable, non-invasive and radiation-free planning of palatal tissue harvesting and can also be used for location of the GPA at 85% of measurement points, which might help reduce complications during surgery.

Determination of the palatal masticatory mucosa thickness by dental MRI: a prospective study analysing age and gender effects.

OBJECTIVES: The aim of this prospective study was to evaluate age and gender effects on the palatal masticatory mucosa thickness by using non-invasive and non-ionizing MRI. METHODS: 40 periodontally healthy participants of five gender-balanced age groups (20-29, 30-39, 40-49, 50-59 and 60-69 years, respectively) underwent dental MRI at 3 Tesla using a contrast-enhanced, high-resolution 3D-sequence. The palatal masticatory mucosa was measured at 40 standard measurement points by two independent observers. Statistical analysis was performed by using intraclass correlation coefficient (ICC), Shapiro-Wilk test, two-way analysis of variance and post-hoc Tukey HSD test. RESULTS: Measurements of the palatal masticatory mucosa thickness were highly reliable with a mean intraobserver ICC of 0.989 and a mean interobserver ICC of 0.987. Mean palatal masticatory mucosa thickness increased with the distance from the gingival margin in all tooth regions. Molars showed a considerably lower average palatal masticatory mucosa thickness at intermediate heights in comparison to canines and premolars. Average palatal masticatory mucosa thickness continuously increased with age and significantly differed between age groups (p < 0.01). A significant increase was observed between the age groups of 30-39 years and 40-49 years (p = 0.04). Gender had no significant effect on average palatal mucosa thickness (p = 0.19). However, there was a tendency towards thicker mucosa in males (mean ± SD, 3.36 mm ± 0.47) compared to females (3.23 mm ± 0.44). CONCLUSIONS: In the present study, dental MRI allowed for a highly reliable determination of the palatal masticatory mucosa thickness. Considerable intra- and interindividual variations in palatal masticatory mucosa thickness were observed. Average palatal masticatory mucosa thickness was dependent on age but not on gender.

PETRA, MSVAT-SPACE and SEMAC sequences for metal artefact reduction in dental MR imaging.

OBJECTIVES: Dental MRI is often impaired by artefacts due to metallic dental materials. Several sequences were developed to reduce susceptibility artefacts. Here, we evaluated a set of sequences for artefact reduction for dental MRI for the first time. METHODS: Artefact volume, signal-to-noise ratio (SNR) and image quality were assessed on a 3-T MRI for pointwise encoding time reduction with radial acquisition (PETRA), multiple-slab acquisition with view angle tilting gradient, based on a sampling perfection with application-optimised contrasts using different flip angle evolution (SPACE) sequence (MSVAT-SPACE), slice-encoding for metal-artefact correction (SEMAC) and compared to a standard SPACE and a standard turbo-spin-echo (TSE) sequence. Field-of-view and acquisition times were chosen to enable in vivo application. Two implant-supported prostheses were tested (porcelain fused to metal non-precious alloy and monolithic zirconia). RESULTS: Smallest artefact was measured for TSE sequences with no difference between the standard TSE and the SEMAC. MSVAT-SPACE reduced artefacts about 56% compared to the standard SPACE. Effect of the PETRA was dependent on sample used. Image quality and SNR were comparable for all sequences except PETRA, which yielded poor results. CONCLUSION: There is no benefit in terms of artefact reduction for SEMAC compared to standard TSE. Usage of MSVAT-SPACE is advantageous since artefacts are reduced and higher resolution is achieved. KEY POINTS: • SEMAC is not superior to TSE in terms of artefact reduction. • MSVAT-SPACE reduces susceptibility artefacts while maintaining comparable image quality. • PETRA reduces susceptibility artefacts depending on material but offers poor image quality.

Lateral cephalometric analysis for treatment planning in orthodontics based on MRI compared with radiographs: A feasibility study in children and adolescents.

OBJECTIVE: The objective of this prospective study was to evaluate whether magnetic resonance imaging (MRI) is equivalent to lateral cephalometric radiographs (LCR, "gold standard") in cephalometric analysis. METHODS: The applied MRI technique was optimized for short scanning time, high resolution, high contrast and geometric accuracy. Prior to orthodontic treatment, 20 patients (mean age ± SD, 13.95 years ± 5.34) received MRI and LCR. MRI datasets were postprocessed into lateral cephalograms. Cephalometric analysis was performed twice by two independent observers for both modalities with an interval of 4 weeks. Eight bilateral and 10 midsagittal landmarks were identified, and 24 widely used measurements (14 angles, 10 distances) were calculated. Statistical analysis was performed by using intraclass correlation coefficient (ICC), Bland-Altman analysis and two one-sided tests (TOST) within the predefined equivalence margin of ± 2°/mm. RESULTS: Geometric accuracy of the MRI technique was confirmed by phantom measurements. Mean intraobserver ICC were 0.977/0.975 for MRI and 0.975/0.961 for LCR. Average interobserver ICC were 0.980 for MRI and 0.929 for LCR. Bland-Altman analysis showed high levels of agreement between the two modalities, bias range (mean ± SD) was -0.66 to 0.61 mm (0.06 ± 0.44) for distances and -1.33 to 1.14° (0.06 ± 0.71) for angles. Except for the interincisal angle (p = 0.17) all measurements were statistically equivalent (p < 0.05). CONCLUSIONS: This study demonstrates feasibility of orthodontic treatment planning without radiation exposure based on MRI. High-resolution isotropic MRI datasets can be transformed into lateral cephalograms allowing reliable measurements as applied in orthodontic routine with high concordance to the corresponding measurements on LCR.

Artefacts of implant-supported single crowns - Impact of material composition on artefact volume on dental MRI.

PURPOSE: MRI allows radiation-free imaging of the head and neck area. However, implant-supported prostheses may severely impair image quality due to artefacts. Therefore, identification of preferable material compositions for implants and supported prostheses with little impact on MR image quality is mandatory. MATERIALS AND METHODS: Overall, one zirconia and four titanium dental implants were provided with different single crown materials: porcelain-fused-to-metal precious alloy (GP-T), porcelain-fused-to-metal non-precious alloy (CCT-T), porcelain-fused-to-zirconia (ZC-T) and monolithic zirconia (Z-T, Z-Z). Three-dimensional artefact volume was determined on a 3 Tesla MRI, applying two standard sequence types (SPACE and TSE). Two-way ANOVA and pair-wise post-hoc Turkey test were performed for comparison of artefact size. RESULTS: Fewest MR artefacts were observed with zirconia implant combined with monolithic zirconia crown. A titanium implant combined with a single crown framework out of the non-precious alloys was unfavourable in terms of artefact volume. Smaller and comparable artefact volumes were noted for titanium implants with the remaining three crown materials (GP-T, ZC-T and Z-T). CONCLUSIONS: Material composition of dental implants provided with single crowns has a profound impact on artefact volume. In comparison with crowns containing cobalt, chromium and tungsten, the MRI artefacts are reduced in precious alloy- and zirconia-based crowns. Further studies are needed to assess whether residual artefacts allow sufficient diagnostic imaging with these crowns. Conflict-of-interest statement: The authors have nothing to disclose.

Superior in vitro stimulation of human CD8+ T-cells by whole virus versus split virus influenza vaccines.

Pandemic and seasonal influenza viruses cause considerable morbidity and mortality in the general human population. Protection from severe disease may result from vaccines that activate antigen-presenting DC for effective stimulation of influenza-specific memory T cells. Special attention is paid to vaccine-induced CD8+ T-cell responses, because they are mainly directed against conserved internal influenza proteins thereby presumably mediating cross-protection against circulating seasonal as well as emerging pandemic virus strains. Our study showed that influenza whole virus vaccines of major seasonal A and B strains activated DC more efficiently than those of pandemic swine-origin H1N1 and pandemic-like avian H5N1 strains. In contrast, influenza split virus vaccines had a low ability to activate DC, regardless which strain was investigated. We also observed that whole virus vaccines stimulated virus-specific CD8+ memory T cells much stronger compared to split virus counterparts, whereas both vaccine formats activated CD4+ Th cell responses similarly. Moreover, our data showed that whole virus vaccine material is delivered into the cytosolic pathway of DC for effective activation of virus-specific CD8+ T cells. We conclude that vaccines against seasonal and pandemic (-like) influenza strains that aim to stimulate cross-reacting CD8+ T cells should include whole virus rather than split virus formulations.