The novel mTOR inhibitor RAD001 (Everolimus) induces antiproliferative effects in human pancreatic neuroendocrine tumor cells

Background/Aim: Tumors exhibiting constitutively activated PI(3) K/Akt/mTOR signaling are hypersensitive to mTOR inhibitors such as RAD001 (everolimus) which is presently being investigated in clinical phase II trials in various tumor entities, including neuroendocrine tumors (NETs). However, no preclinical data about the effects of RAD001 on NET cells have been published. In this study, we aimed to evaluate the effects of RAD001 on BON cells, a human pancreatic NET cell line that exhibits constitutively activated PI(3) K/Akt/mTOR signaling. Methods: BON cells were treated with different concentrations of RAD001 to analyze its effect on cell growth using proliferation assays. Apoptosis was examined by Western blot analysis of caspase-3/PARP cleavage and by FACS analysis of DNA fragmentation. Results: RAD001 potently inhibited BON cell growth in a dose-dependent manner which was dependent on the serum concentration in the medium. RAD001-induced growth inhibition involved G0/G1-phase arrest as well as induction of apoptosis. Conclusion: In summary, our data demonstrate antiproliferative and apoptotic effects of RAD001 in NET cells in vitro supporting its clinical use in current phase II trials in NET patients. Copyright (c) 2007 S. Karger AG, Basel

A microtubule interactome: complexes with roles in cell cycle and mitosis.

addresses: Department of Zoology, University of Oxford, Oxford, United Kingdom.notes: PMCID: PMC2323305types: Journal Article; Research Support, Non-U.S. Gov'tCopyright: © 2008 Hughes et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.The microtubule (MT) cytoskeleton is required for many aspects of cell function, including the transport of intracellular materials, the maintenance of cell polarity, and the regulation of mitosis. These functions are coordinated by MT-associated proteins (MAPs), which work in concert with each other, binding MTs and altering their properties. We have used a MT cosedimentation assay, combined with 1D and 2D PAGE and mass spectrometry, to identify over 250 MAPs from early Drosophila embryos. We have taken two complementary approaches to analyse the cellular function of novel MAPs isolated using this approach. First, we have carried out an RNA interference (RNAi) screen, identifying 21 previously uncharacterised genes involved in MT organisation. Second, we have undertaken a bioinformatics analysis based on binary protein interaction data to produce putative interaction networks of MAPs. By combining both approaches, we have identified and validated MAP complexes with potentially important roles in cell cycle regulation and mitosis. This study therefore demonstrates that biologically relevant data can be harvested using such a multidisciplinary approach, and identifies new MAPs, many of which appear to be important in cell division

Peri-implant diseases: Consensus Report of the Sixth European Workshop on Periodontology

Issues related to peri-implant disease were discussed. It was observed that the most common lesions that occur, i.e. peri-implant mucositis and peri-implantitis are caused by bacteria. While the lesion of peri-implant mucositis resides in the soft tissues, peri-implantitis also affects the supporting bone. Peri-implant mucositis occurs in about 80% of subjects (50% of sites) restored with implants, and peri-implantitis in between 28% and 56% of subjects (12-40% of sites). A number of risk indicators were identified including (i) poor oral hygiene, (ii) a history of periodontitis, (iii) diabetes and (iv) smoking. It was concluded that the treatment of peri-implant disease must include anti-infective measures. With respect to peri-implant mucositis, it appeared that non-surgical mechanical therapy caused the reduction in inflammation (bleeding on probing) but also that the adjunctive use of antimicrobial mouthrinses had a positive effect. It was agreed that the outcome of non-surgical treatment of peri-implantitis was unpredictable. The primary objective of surgical treatment in peri-implantitis is to get access to the implant surface for debridement and decontamination in order to achieve resolution of the inflammatory lesion. There was limited evidence that such treatment with the adjunctive use of systemic antibiotics could resolve a number of peri-implantitis lesions. There was no evidence that so-called regenerative procedures had additional beneficial effects on treatment outcome

Comparative analysis of the lambda-interferons IL-28A and IL-29 regarding their transcriptome and their antiviral properties against hepatitis C virus.

Specific differences in signaling and antiviral properties between the different Lambda-interferons, a novel group of interferons composed of IL-28A, IL-28B and IL-29, are currently unknown. This is the first study comparatively investigating the transcriptome and the antiviral properties of the Lambda-interferons IL-28A and IL-29. Expression studies were performed by microarray analysis, quantitative PCR (qPCR), reporter gene assays and immunoluminometric assays. Signaling was analyzed by Western blot. HCV replication was measured in Huh-7 cells expressing subgenomic HCV replicon. All hepatic cell lines investigated as well as primary hepatocytes expressed both IFN-λ receptor subunits IL-10R2 and IFN-λR1. Both, IL-28A and IL-29 activated STAT1 signaling. As revealed by microarray analysis, similar genes were induced by both cytokines in Huh-7 cells (IL-28A: 117 genes; IL-29: 111 genes), many of them playing a role in antiviral immunity. However, only IL-28A was able to significantly down-regulate gene expression (n = 272 down-regulated genes). Both cytokines significantly decreased HCV replication in Huh-7 cells. In comparison to liver biopsies of patients with non-viral liver disease, liver biopsies of patients with HCV showed significantly increased mRNA expression of IL-28A and IL-29. Moreover, IL-28A serum protein levels were elevated in HCV patients. In a murine model of viral hepatitis, IL-28 expression was significantly increased. IL-28A and IL-29 are up-regulated in HCV patients and are similarly effective in inducing antiviral genes and inhibiting HCV replication. In contrast to IL-29, IL-28A is a potent gene repressor. Both IFN-λs may have therapeutic potential in the treatment of chronic HCV

Additive Anti-Tumor Effects of Lovastatin and Everolimus In Vitro through Simultaneous Inhibition of Signaling Pathways

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are creditedThis work was supported by a research grant from the Ludwig-Maximilians University of Munich (Förderprogramm für Forschung und Lehre [FöFoLe], grant number 865/829)

Síndrome de Stevens-Johnson. Apresentação de Caso Clínico

Introdução: A Síndrome de Stevens-Johnson (SSJ) é uma doença mucocutânea rara e potencialmente fatal, mais frequente no sexo masculino, cuja incidência aumenta com a idade e em determinados grupos de risco. A SSJ e a Necrólise Tóxica Epidérmica (NET) são duas entidades da mesma doença, com severidade diferente. A etiologia não é clara, mas pensa-se que se deva maioritariamente a reacções adversas a fármacos. Caso clínico: Um jovem de 17 anos de idade, sem antecedentes pessoais relevantes, foi observado no Serviço de Urgência por surgimento de lesões maculopapulares, com 3 dias de evolução, dispersas pela face, cavidade oral, tronco e extremidades, com prostração e taquicardia. Foi internado com o diagnóstico de SSJ. Discussão e Conclusões: O SSJ e a NET têm grande morbilidade e considerável mortalidade. O rápido reconhecimento desta identidade, com a remoção do fármaco desencadeador é essencial. A perda da função de barreira da pele, com a consequente alteração da homeostasia, implica muitas vezes a manutenção da terapêutica de suporte em Unidades de Cuidados Intensivos ou de Queimados.info:eu-repo/semantics/publishedVersio

Feasibility Testing of the Automatic Design of Three-Unit Implant Fixed Dental Prostheses with Different Dental CAD Software: A Pre-Clinical Pilot Trial

Background/Objectives: The technical development of implant-supported fixed dental prostheses (iFDP) initially concentrated on the computer-aided manufacturing of prosthetic restorations (CAM). Advances in information technologies have shifted the focus for optimizing digital workflows to AI-based processes for design (CAD). This pre-clinical pilot trial investigated the feasibility of the automatic design of three-unit iFDPs using CAD software (Dental Manger 2021, 3Shape; DentalCAD 3.1 Rijeka, exocad GmbH). Methods: Two clinical scenarios based on a full dentition were created virtually. Physical models were produced and digitized using two intraoral scanners applying quadrant or full-arch scans (Trios3, 3Shape, Copenhagen, Denmark; and Primescan AC, Dentsply Sirona, Bensheim, Germany). For each scenario, iFDP designs were generated automatically using two laboratory software systems (Dental Manger 2021, 3Shape; DentalCAD 3.1 Rijeka, exocad GmbH), resulting in 80 STL datasets (2 scenarios × 2 scan strategies × 2 IOS systems × 5 scan repetitions × 2 software). The files were analyzed clinically for the contact schemes and pontic area. One of the automated designs for each scenario was manually post-processed and one iFDP design for each scenario was manually created by experienced dental technicians (control). The time required for all the design processes was recorded. Results: The automatic design of iFDPs without manual adjustment did not lead to clinically acceptable restorations. The time required for the automatically generated/manually adjusted iFDPs designs was not significantly different to that for the manually designed restorations. Conclusions: Current laboratory software can not automatically generate three-unit iFDPs with clinically acceptable results in terms of the interproximal and occlusal contacts and the pontic design. The automatic iFDP design process currently requires manual adjustment, which means there is no benefit in terms of the working time compared with manually created restorations

In planta preliminary screening of ER glycoprotein folding quality control (ERQC) modulators

Small molecule modulators of the Endoplasmic Reticulum glycoprotein folding quality control (ERQC) machinery have broad-spectrum antiviral activity against a number of enveloped viruses and have the potential to rescue secretion of misfolded but active glycoproteins in rare diseases. In vivo assays of candidate inhibitors in mammals are expensive and cannot be afforded at the preliminary stages of drug development programs. The strong conservation of the ERQC machinery across eukaryotes makes transgenic plants an attractive system for low-cost, easy and fast proof-of-concept screening of candidate ERQC inhibitors. The Arabidopsis thaliana immune response is mediated by glycoproteins, the folding of which is controlled by ERQC. We have used the plant response to bacterial peptides as a means of assaying an ERQC inhibitor in vivo. We show that the treatment of the plant with the iminosugar NB-DNJ, which is a known ER α-glucosidase inhibitor in mammals, influences the immune response of the plant to the bacterial peptide elf18 but not to the flagellin-derived flg22 peptide. In the NB-DNJ-treated plant, the responses to elf18 and flg22 treatments closely follow the ones observed for the ER α-glucosidase II impaired plant, At psl5-1. We propose Arabidopsis thaliana as a promising platform for the development of low-cost proof-of-concept in vivo ERQC modulation

Precision of maxillo-mandibular registration with intraoral scanners in vitro

Purpose: To compare the precision of maxillo-mandibular registration and resulting full arch occlusion produced by three intraoral scanners in vitro. Methods: Six dental models (groups A–F) were scanned five times with intraoral scanners (CEREC, TRIOS, PLANMECA), producing both full arch and two buccal maxillo-mandibular scans. Total surface area of contact points (defined as regions within 0.1 mm and all mesh penetrations) was measured, and the distances between four pairs of key points were compared, each two in the posterior and anterior. Results: Total surface area of contact points varied significantly among scanners across all groups. CEREC produced the smallest contact surface areas (5.7–25.3 mm2), while PLANMECA tended to produce the largest areas in each group (22.2–60.2 mm2). Precision of scanners, as measured by the 95% CI range, varied from 0.1–0.9 mm for posterior key points. For anterior key points the 95% CI range was smaller, particularly when multiple posterior teeth were still present (0.04–0.42 mm). With progressive loss of posterior units (groups D–F), differences in the anterior occlusion among scanners became significant in five out of six groups (D–F left canines and D, F right canines, p < 0.05). Conclusions: Maxillo-mandibular registrations from three intraoral scanners created significantly different surface areas of occlusal contact. Posterior occlusions revealed lower precision for all scanners than anterior. CEREC tended towards incorrect posterior open bites, whilst TRIOS was most consistent in reproducing occluding units

Towards the first antibody-free serum biomarker assay for NAFLD

Liver biopsy is the reference standard for NAFLD diagnosis and staging. NAFLD can be assessed less invasively using immunoassays to detect serum biomarkers. However, biomarkers can degrade due to sample storage conditions and therefore may not be detected using these antibody-based assays. Detection of biomarkers by mass spectrometry (MS) overcomes this disadvantage