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.

A modular synthetic route to size-defined immunogenic Haemophilus influenzae b antigens is key to the identification of an octasaccharide lead vaccine candidate.

The first glycoconjugate vaccine using isolated glycans was licensed to protect children from Haemophilus influenzae serotype b (Hib) infections. Subsequently, the first semisynthetic glycoconjugate vaccine using a mixture of antigens derived by polymerization targeted the same pathogen. Still, a detailed understanding concerning the correlation between oligosaccharide chain length and the immune response towards the polyribosyl-ribitol-phosphate (PRP) capsular polysaccharide that surrounds Hib remains elusive. The design of semisynthetic and synthetic Hib vaccines critically depends on the identification of the minimally protective epitope. Here, we demonstrate that an octasaccharide antigen containing four repeating disaccharide units resembles PRP polysaccharide in terms of immunogenicity and recognition by anti-Hib antibodies. Key to this discovery was the development of a modular synthesis that enabled access to oligosaccharides up to decamers. Glycan arrays containing the synthetic oligosaccharides were used to analyze anti-PRP sera for antibodies. Conjugates of the synthetic antigens and the carrier protein CRM197, which is used in licensed vaccines, were employed in immunization studies in rabbits.

Transcriptomic and proteomic insight into the effects of a defined European mistletoe extract in Ewing sarcoma cells reveals cellular stress responses.

BACKGROUND: The hydrophobic triterpenes, oleanolic and betulinic acid as well as the hydrophilic mistletoe lectins and viscotoxins possess anticancer properties. They do all occur in combination in European mistletoe (Viscum album L.). Commercial Viscum album L. extracts are aqueous, excluding the insoluble triterpenes. We have previously shown that mistletoe lectins and triterpene acids are effective against Ewing sarcoma in vitro, ex vivo and in vivo. METHODS: We recreated a total mistletoe effect (viscumTT) by combining an aqueous extract (viscum) and a triterpene extract (TT) solubilised with cyclodextrins and analysed the effects of viscumTT and the single extracts on TC-71 Ewing sarcoma cells in vitro by transcriptomic and proteomic profiling. RESULTS: Treatment with the extracts strongly impacted Ewing sarcoma cell gene and protein expression. Apoptosis-associated and stress-activated genes were upregulated, proteasomal protein abundance enhanced and ribosomal and spliceosomal proteins downregulated. The mechanism of action of viscum, TT and viscumTT in TC-71 and MHH-ES-1 cells suggests the involvement of the unfolded protein response. While viscum and viscumTT extract treatment indicate response to oxidative stress and activation of stress-mediated MAPK signalling, TT extract treatment suggests the involvement of TLR signalling and autophagy. CONCLUSIONS: Since the combinatory extract viscumTT exerts highly effective pro-apoptotic effects on Ewing sarcoma cells in vitro, this phytopolychemotherapy could be a promising adjuvant therapeutic option for paediatric patients with Ewing sarcoma.

Mitochondrial DNA mutations in renal cell carcinomas revealed no general impact on energy metabolism.

Previously, renal cell carcinoma tissues were reported to display a marked reduction of components of the respiratory chain. To elucidate a possible relationship between tumourigenesis and alterations of oxidative phosphorylation, we screened for mutations of the mitochondrial DNA (mtDNA) in renal carcinoma tissues and patient-matched normal kidney cortex. Seven of the 15 samples investigated revealed at least one somatic heteroplasmic mutation as determined by denaturating HPLC analysis (DHPLC). No homoplasmic somatic mutations were observed. Actually, half of the mutations presented a level of heteroplasmy below 25%, which could be easily overlooked by automated sequence analysis. The somatic mutations included four known D-loop mutations, four so far unreported mutations in ribosomal genes, one synonymous change in the ND4 gene and four nonsynonymous base changes in the ND2, COI, ND5 and ND4L genes. One renal cell carcinoma tissue showed a somatic A3243G mutation, which is a known frequent cause of MELAS syndrome (mitochondrial encephalomyopathy, lactic acidosis, stroke-like episode) and specific compensatory alterations of enzyme activities of the respiratory chain in the tumour tissue. No difference between histopathology and clinical progression compared to the other tumour tissues was observed. In conclusion, the low abundance as well as the frequently observed low level of heteroplasmy of somatic mtDNA mutations indicates that the decreased aerobic energy capacity in tumour tissue seems to be mediated by a general nuclear regulated mechanism.