TRAIL/TRAIL Receptor System and Susceptibility to Multiple Sclerosis

The TNF-related apoptosis inducing ligand (TRAIL)/TRAIL receptor system participates in crucial steps in immune cell activation or differentiation. It is able to inhibit proliferation and activation of T cells and to induce apoptosis of neurons and oligodendrocytes, and seems to be implicated in autoimmune diseases. Thus, TRAIL and TRAIL receptor genes are potential candidates for involvement in susceptibility to multiple sclerosis (MS). To test whether single-nucleotide polymorphisms (SNPs) in the human genes encoding TRAIL, TRAILR-1, TRAILR-2, TRAILR-3 and TRAILR-4 are associated with MS susceptibility, we performed a candidate gene case-control study in the Spanish population. 59 SNPs in the TRAIL and TRAIL receptor genes were analysed in 628 MS patients and 660 controls, and validated in an additional cohort of 295 MS patients and 233 controls. Despite none of the SNPs withstood the highly conservative Bonferroni correction, three SNPs showing uncorrected p values<0.05 were successfully replicated: rs4894559 in TRAIL gene, p = 9.8×10−4, OR = 1.34; rs4872077, in TRAILR-1 gene, p = 0.005, OR = 1.72; and rs1001793 in TRAILR-2 gene, p = 0.012, OR = 0.84. The combination of the alleles G/T/A in these SNPs appears to be associated with a reduced risk of developing MS (p = 2.12×10−5, OR = 0.59). These results suggest that genes of the TRAIL/TRAIL receptor system exerts a genetic influence on MS

The deimination of myelin basic protein as a post-translational modification contributing to the pathogenesis of demyelinating disease

grantor: University of TorontoMyelin basic protein (MBP) consists of a family of charge isomers generated by post-translational modifications. Deimination is one such modification which results in the conversion of arginyl residues to citrulline producing a net loss of positive charge within MBP. The enzyme considered to be responsible for the deimination of proteins is peptidylarginine deiminase (PAD). Citrullinated MBP has been found to be the major charge isomer present early in development (in infants up to 2 years). An increased amount of citrullinated MBP has also been noted in both the brains of patients with MS and in the brains of ND4 mice (an animal model of demyelinating disease). Citrullinated MBP has been found to be more susceptible to proteolytic digestion by cathepsin D such that the rate of digestion increased 0 citrulline < 6 citrulline < 18 citrulline. In this study, MBP containing various numbers of citrullinyl residues was generated using PAD 'in vitro' and the rate of digestion slowly increased at low numbers of citrulline (less than 4) but increased rapidly at 7 mol citrulline/mol protein. The effect of methylation at arginine 106 (bovine sequence) on deimination was examined by mass spectroscopy. Methylated MBP was found to be less susceptible to deimination by PAD ' in vitro' and citrullinated MBP isolated from MS brains was found to be less methylated suggesting that methylation of arginine 106 (107 of human sequence) may protect against deimination. Methylation of arginine 107 (human sequence) was found to result in a compaction of the molecule (as shown by molecular modelling) perhaps rendering MBP less accessible to PAD. In this study, the developmental changes in PAD and citrullinated MBP were studied in normal and ND4 mice. The amount of citrullinated MBP was affected by the amount of PAD but this relationship appeared to be subject to a number of other influences such as myelin compaction, methylation of arginine 106 and the increased rate of proteolytic digestion of citrullinated MBP. In order to decrease the amount of citrullinated MBP generated in demyelinating disease, an inhibitor of PAD activity 'in vitro' (paclitaxel) was identified.Ph.D

RNA disruption and drug response in breast cancer primary systemic therapy

BACKGROUND: As there is now evidence that switching clinical nonresponders early in primary systemic therapy to alternate treatment regimens can enhance survival in some breast cancer patients, the need for a robust intermediate endpoint that can guide treatment response across all tumor subtypes is urgent. Recently, chemotherapy drugs have been shown to induce a decrease in RNA quality in tumor cells from breast cancer biopsies in some patients at midtherapy, and that this has been associated with subsequent achievement of pathological complete response (pCR). The decrease in RNA quality has been shown to be associated with RNA disruption; aberrant RNA bands visualized by RNA electrophoresis have been associated with subsequent tumor cell death. The objectives of these studies are to show that a new assay based on induction of RNA disruption in tumor cells by chemotherapy can stratify at midtherapy, pCR responders from non-pCR responders irrespective of clinical response and to present early evidence that clinically useful RNA disruption can be detected as early as 14 days after initiation of treatment. METHODS: RNA disruption in tumor cells was quantified by analysis of the RNA electrophoresis banding pattern and expressed as an RNA disruption index (RDI). To develop the RNA disruption assay (RDA), RDI was correlated with clinical outcome (pCR) from the NCIC-CTG MA.22 breast cancer clinical trial (ClinicalTrials.gov NCT00066443). RDA Zones were established by stratifying patients using RDI values into Zone 1, Zone 2, and Zone 3. Zone 3 included seven out of eight pCR responders, whereas Zone 1 contained no pCR responders. An intermediate zone (Zone 2) was established which contained one pCR. Subsequently, to determine early drug response, RNA disruption was examined by RDI after 14 days exposure to trastuzumab, zoledronic acid, or letrozole + cyclophosphamide \ub1 sorafenib therapy. RESULTS: In MA.22, RDA stratified 23 of 85 patients in Zone 1 as pCR nonresponders, 24 patients in Zone 2, an intermediate zone, and 38 patients in Zone 3, pCR responders and non-pCR patients who share RDI comparable to those achieving pCR. In the early response studies, after 14 days exposure to chemotherapy, some RNA disruption as measured by RDI elevation could be detected in 3/12 trastuzumab, 7/15 zoledronic acid, 5/29 letrozole + cyclophosphamide, and 5/23 letrozole + cyclophosphamide + sorafenib patients. CONCLUSIONS: RDA is a novel intermediate endpoint that has promise for clinical utility for breast cancers early in response-guided primary systemic therapy

How do school principals assign teachers to classes?

The education system in the Czech Republic allows sorting pupils according to their abilities in the first and second level of primary school. Teachers are therefore placed to pupils in selective or non-selective classes and groups by headmasters and management of primary schools. This thesis deals with the placing of teachers to classes by headmasters and management in primary schools and aims to determine by what way and according to which criteria teachers are placed to different types of classes. The research in this thesis is supported by studies, which deal with the quality of teacher and differential education since the 60th of the 20th century. The theoretical part of the thesis focuses on the definition of terms related to differential education and studies how teachers and the teaching are viewed. The empirical part describes the research methodology of qualitative research which objective is to discover how the headmasters or management place teachers to classes, in addition what criteria are decisive for them and how they define qualities of the placed teachers. Key words: External differentiation, internal differentiation, quality of teacher, teaching approaches selective primary school, the sorting of students, abilities, Principal, the placing of teachers, selective classes,..

Additional file 4: of RNA disruption is associated with response to multiple classes of chemotherapy drugs in tumor cell lines

Northern blots of total A2780 RNA hybridized with 18S probes. Panels on the left side show an RNA gel and panels on the right side show corresponding membranes after transfer and blotting. RNA isolated from untreated cells is in lanes marked (-) while RNA isolated from 48 h 0.2 ΟM docetaxel treated cells is in lanes marked (+). Sizes of RNA bands on gels are shown in nucleotide length (nt) to the left of the figure with arrows indicating each band. Sizes of hybridization targets of probes are shown in nucleotides to the right of the figures with arrows indicating each hybridization target. A. Northern blot results using the 18S-1 probe. B. Northern blot results using the 18S-2 probe. C. Northern blot results using the 18S-3 probe. (PPTX 410 kb

RNA disruption is a widespread phenomenon associated with stress-induced cell death in tumour cells

Abstract We have previously shown that neoadjuvant chemotherapy can induce the degradation of tumour ribosomal RNA (rRNA) in patients with advanced breast cancer, a phenomenon we termed “RNA disruption”. Extensive tumour RNA disruption during chemotherapy was associated with a post-treatment pathological complete response and improved disease-free survival. The RNA disruption assay (RDA), which quantifies this phenomenon, is now being evaluated for its clinical utility in a large multinational clinical trial. However, it remains unclear if RNA disruption (i) is manifested across many tumour and non-tumour cell types, (ii) can occur in response to cell stress, and (iii) is associated with tumour cell death. In this study, we show that RNA disruption is induced by several mechanistically distinct chemotherapy agents and report that this phenomenon is observed in response to oxidative stress, endoplasmic reticulum (ER) stress, protein translation inhibition and nutrient/growth factor limitation. We further show that RNA disruption is dose- and time-dependent, and occurs in both tumourigenic and non-tumourigenic cell types. Northern blotting experiments suggest that the rRNA fragments generated during RNA disruption stem (at least in part) from the 28S rRNA. Moreover, we demonstrate that RNA disruption is reproducibly associated with three robust biomarkers of cell death: strongly reduced cell numbers, lost cell replicative capacity, and the generation of cells with a subG1 DNA content. Thus, our findings indicate that RNA disruption is a widespread phenomenon exhibited in mammalian cells under stress, and that high RNA disruption is associated with the onset of cell death

Pure anti-tumor effect of zoledronic acid in na\uefve bone-only metastatic and locally advanced breast cancer: Proof from the "biological window therapy"

The study investigated the anti-tumour effect of zoledronic acid (ZA) administered alone in a biological window therapy in na\uefve bone-only metastatic and locally advanced breast cancer (LABC) patients. 33 patients with LABC (Group 1) and 20 patients with a first diagnosis of bone metastasis only (Group 2) received 4 mg single dose of ZA, 14 days (biological window) before starting any treatment. In Group 1, Ki67, CD34, p53/bcl-2 and caspase 3 expression along with the adenosine triphosphate (ATP) levels and RNA disruption index were evaluated as markers of tumor growth in tumour specimens obtained before and after ZA administration (basal, day 14). In Group 2, the total enumeration of circulating tumour cells (CTCs), and of M30+ve CTCs along with the soluble marker of cell death (M30/M65) were carried-out as markers of tumor dissemination at baseline, at 48 h and day 14th. In Group 1, there was a significant reduction in Ki67, CD34, bcl-2 expression after 14 days ZA based-treatment (p = 0.0032; p = 0.0001, p < 0.00001 respectively). ZA showed a significant increase of RNA disruption (p < 0.0076). In Group 2, we observed a significant reduction of CTCs number after 48 h (p = 0.0012), followed by a significant rebound at 14 days (p = 0.012). The apoptotic CTCs/M30+ve and M65 levels significantly increased under treatment (p = 0.018 and p = 0.039 respectively) after drug administration when compared to the baseline. These results are the first prospective in vivo data showing the direct pure anti-tumour effect (either on the tumour cell or on CTCs) of ZA