Lysyl oxidase like-2 mediates tumor to stromal cell communication in oral cancer

INTRODUCTION: The lysyl oxidase family consists of 5 members and oxidizes specific lysine residues in biosynthetic collagen and elastin maturation. Lysyl oxidase like-2 (LOXL2) is elevated in oral cancer and promotes metastasis and correlates with poor prognosis. The objective of this study was to determine the mechanism by which LOXL2 promotes the progression and invasiveness of oral squamous cell carcinoma. METHODS: In vitro: The effects of LOXL2 inhibitor (PXS-S1C) on human gingival fibroblasts treated with tumor cell conditioned medium (CM) were investigated. Cell proliferation assays, signaling arrays, gene knock down and western blots were used to evaluate the effect of PXS-S1C on CM-treated fibroblasts. The effects of PXS-S1C on cancer cell expression of LOXL2 and proliferation were determined. To find potential LOXL2 substrates, carbonyl-containing proteins of gingival fibroblasts treated with CM +/- PXS-S1C were affinity-labeled and then purified by affinity chromatography and identified by western blot. In vivo: The effects of PXS-S1C on cancer cell growth and metastasis in vivo were investigated using orthotopic oral tongue cancer mouse models in both immunodeficient and immunocompetent mice. PXS-S1C at 10 mg/kg and 30 mg/kg was injected immediately following tumor cell injections. Tumor growth was monitored by both caliper measurement and in vivo imaging (IVIS). The mice were sacrificed and their organs were subjected to immunohistochemical staining with proliferation markers. RESULTS: PXS-S1C significantly inhibited gingival fibroblast proliferation stimulated by tumor cell CM and attenuated phosphorylation of PDGFRβ at the Y771 and Y857, but not Y751 residues in response to CM treatment. PXS-S1C inhibited ERK1/2-signaling in fibroblasts but not AKT pathway in response to CM treatment. PDGFR activation by oral tumor cells was mimicked by PDGF-AB but not PDGF-BB. PXS-S1C decreased the expression of LOXL2 in HSC3 oral cancer cells in vitro, suggesting the existence of a positive autoregulatory loop. Assessing for direct LOXL2 substrates in fibroblasts with functional consequences identified PDGFR. In vivo studies: Caliper measurements, IVIS, and immunohistochemistry demonstrated that inhibition of LOXL2 by injections of PXS-S1C significantly decreased both progression and metastasis of oral cancers in vivo in both mouse models. Mice without PXS-S1C treatment developed larger tongue volumes (p<0.05), and in immunocompetent mice larger lymph nodes (9 out 12) were observed compared to the PXS-S1C-treated mice (4 out of 12). IVIS imaging of immunodeficient mice revealed inhibition of metastasis by PXS-S1C treatment. The expression of proliferation marker (Ki-67 or PCNA) and LOXL2 was lower in tongue tumors treated with PXS-S1C in both in vivo models (p<0.05). CONCLUSIONS: LOXL2 secreted by cancer cells stimulates fibroblast proliferation by oxidizing PDGFR and thereby enhancing PDGF-mediated signaling. Inhibition of LOXL2 can be used as a therapeutic strategy to suppress the growth and metastasis of oral cancers by modulating tumor microenvironment.2019-10-24T00:00:00

Monitoring program for pharmaceuticals, illegal substances, and contaminants in farmed fish - Annual report for 2021

This report summarises the monitoring data collected in 2021 on the status of illegal substances, pharmaceuticals and contaminants in Norwegian farmed fish. In 2021, a total of 14135 fish were sampled. Samples examined for illegal compounds were collected at all stages of farming and are representative of farmed fish under production. The samples were analysed for substances with anabolic effects or unauthorized substances. Residues of the dye crystal violet were detected in two samples of salmon. A follow-up investigation conducted by the Norwegian Food Safety Authority (NSFA) concluded with the cause likely being a contamination of the samples during sample taking. No other residues of illegal compounds were found. Samples tested for approved veterinary drugs and contaminants were collected at processing plants and are representative of Norwegian farmed fish ready for human consumption. Residues of the anti-sealice agents cypermethrin, emamectin and lufenuron were found, with concentrations below the respective Maximum Residue Limits (MRLs). Other veterinary drugs, like antibiotics or drugs used against internal parasites were not found. No environmental contaminants were found above the EU maximum level (ML).Monitoring program for pharmaceuticals, illegal substances, and contaminants in farmed fish - Annual report for 2021publishedVersio

Comparing microleakage in root canals obturated with nanosilver coated gutta-percha to standard gutta-percha by two different methods

INTRODUCTION: Favorable apical seal of root filling materials is a crucial factor for a successful root canal treatment. The aim of this in vitro study was to compare bacterial and dye microleakage of two root canal filling materials including standard gutta-percha and nanosilver coated gutta-percha, and to evaluate the agreement between results of these two methods.MATERIALS & METHODS: Fifty-eight extracted single-rooted teeth were randomly divided into two experimental groups of 26 each, and two control groups of three each. After decoronation, root canals were instrumented by crown-down technique. Obturation was conducted using standard gutta-percha in one of experimental groups and nanosilver-coated gutta-percha in another group. AH26 sealer was used as the sealer in both experimental groups. Bacterial leakage was investigated after 60 days using Enterococcus (E.) faecalis microbial strains, and dye leakage was assessed during 72 hours using 1% methylene blue. The data were statistically analyzed by Chi-square test, Kaplan-Meier survival analysis, and Cohen’s Kappa. RESULTS: There was 84% bacterial leakage in standard gutta-percha group and 76% in nanosilver gutta-percha group. Complete dye leakage occurred in 24% and 27% of standard and nanosilver gutta-percha groups, respectively. The above difference between groups was not significant. In the samples with leakage, recorded times of leakage were not significantly different. There was no significant measure of agreement between dye and bacterial penetration along root-end fillings.CONCLUSION: There was a poor agreement between dye and bacterial leakage methods. Leakage results produced by nanosilver gutta-percha were comparable to those by standard gutta-percha. Considering the antibacterial effects of nanosilver coated gutta-percha, use of this type of gutta-percha might be more efficacious in endodontic treatments.

Bioaccumulation of mercury and transcriptional responses in tusk (Brosme brosme), a deep-water fish from a Norwegian fjord

High concentrations of mercury (Hg) have been documented in deep-water fish species from some Norwegian fjords. In this study, tusk (Brosme brosme) was sampled from four locations in the innermost parts of Sognefjorden in Western Norway. Total Hg and methylmercury (MeHg) levels were measured in liver tissue. To search for potential sublethal effects of Hg, we characterized the hepatic transcriptome in tusk with high and low levels of Hg bioaccumulation using global transcriptomics analysis (RNA-seq). The results showed that there was a significant correlation between fish weight and accumulated concentrations of MeHg but not total Hg. MeHg accounted for 30–40% of total Hg in liver of most of the fish, although at concentrations above 2–3 mg Hg/kg wet weight the percentage of MeHg dropped considerably. Transcriptome analysis resulted in hundreds of differentially expressed genes in the liver of tusk with high Hg levels. Functional enrichment analysis suggested that the top affected pathways are associated with protein folding, adipogenesis, notch signaling, and lipid metabolism (beta-oxidation and phospholipids). Based on transcriptional responses pointing to well-known effects of Hg compounds in fish, the study suggests that tusk in Sognefjorden could be negatively impacted by Hg bioaccumulation.publishedVersio

The Effect of Orthodontic Forces on Tooth Response to Electric Pulp Test

Introduction: The current study investigated the pulp response to electric pulp testing (EPT), before, upon initiation and one month after the start of orthodontic tooth movement. Methods and Materials: A total of 402 anterior teeth from 39 patients (mean age of 16.8±2.7 years) were examined in this non-controlled prospective study. The aligning forces were administered using initial NiTi archwires ligated on fixed appliances by using the MBT straight wire technique. The electrical stimulation was provided by the EPT. The EPT readings were recorded at three time points: before bonding (EPT0), immediately upon initiation (EPT1) and 1 month post-treatment (EPT2). The data were statistically analyzed by the ANOVA and Bonferroni tests (P&lt;0.05). Results: Prior to bonding of the orthodontic brackets, the mean EPT value for all the experimental teeth was 3.42 EPT units. Upon initiation, the mean value of EPT1 for each tooth increased to 7.62 units. One month later, the mean EPT2 values dropped to 6.27 units. At this time point, 64 teeth (16%) of the experimental teeth failed to respond. The differences among EPT values at different time points were significant. There was no association between the EPT values and the location or the type of teeth. Conclusion: The physiological changes in the pulp affect the nerve fibers in the early stages of the orthodontic force application. As a result, thresholds to electrical stimulation would increase and the EPT may not initiate a response. Therefore results obtained by electrical pulp testing should be interpreted accordingly.Keywords:Electric Pulp Test; Orthodontic Treatment; Pulp Vitality

Mercury bioaccumulation pathways in tusk (Brosme brosme) from Sognefjord, Norway: Insights from C and N isotopes

Seafood is the main source of methylmercury (MeHg) exposure for humans and elevated total mercury (Hg) concentrations have been reported in marine fish from Norwegian fjords compared with offshore areas. Hg in tusk fillets (n = 201) and liver samples (n = 177) were measured in individuals from different habitats including offshore, coastal area, outer and inner Sognefjord. Specifically, the effects of habitat, energy sources and trophic complexity on Hg bioaccumulation pathways in tusk (Brosme brosme) were investigated using stable isotopes of carbon (δ13C) and nitrogen (δ15N). The concentrations of Hg in tusk increased from offshore towards inner Sognefjord. While Hg concentrations in sediment were at background levels, tusk fillet samples from 7 of 8 sites in Sognefjord had higher Hg levels than the maximum level set by European Union. Based on these findings, human consumption advice for tusk from Sognefjord was issued by the Norwegian Food Safety Authority. δ13C values in tusk successfully discriminated individuals from different habitats and were positively correlated to Hg concentrations in tusk across individuals, sites and habitats, outlining the potential importance of terrestrial carbon and most likely the atmospheric deposition of Hg from the catchment to the overall Hg bioaccumulation and exposure regime in tusk. Additionally, we postulate that the effects of terrestrial carbon sources increased towards inner Sognefjord and likely influenced Hg bioavailability throughout the food web. In contrast, δ15N values were patchy throughout the fjord system and although trophic position explained some of the Hg variation between individual fish, it was not correlated with Hg variation across sites and habitats. Our results suggest that tusk can accumulate high levels of Hg in fjord ecosystems and that catchment runoff is likely an important driver of Hg bioaccumulation in this species.publishedVersio