A Laboratory Excitation Technique to Test Road Bike Vibration Transmission

This paper describes a technique designed to measure the in-situ acceleration signals that will be used to drive a road simulator in the study of road bike vibration transmission in a laboratory setting. To measure the signals, a bike mounted by a cyclist and towed by a motor vehicle is used. A road simulator using actuators driven by a digital signal is described. The impulse response of the bike used to measure road data is convoluted with the road acceleration in order to obtain the required actuator signal. The reproduction capacity of the simulator is evaluated by comparing the frequency content as well as the time statistical parameters of the acceleration signal measurement with road to the acceleration obtained on the simulator. On a granular road with a broadband excitation spectrum, the vertical excitation obtained with the simulator adequately mimics the measured road acceleration. This technique can be used to compare vibration transmission characteristics among different road bikes.The authors gratefully acknowledge financial support from the National Science and Engineering Council of Canada (NSERC) and the participation of Cervélo and Vroomen-White Design

The Crystal Structure of Human Tyrosyl-DNA Phosphodiesterase, Tdp1

AbstractTyrosyl-DNA phosphodiesterase (Tdp1) catalyzes the hydrolysis of a phosphodiester bond between a tyrosine residue and a DNA 3′ phosphate. The enzyme appears to be responsible for repairing the unique protein-DNA linkage that occurs when eukaryotic topoisomerase I becomes stalled on the DNA in the cell. The 1.69 Å crystal structure reveals that human Tdp1 is a monomer composed of two similar domains that are related by a pseudo-2-fold axis of symmetry. Each domain contributes conserved histidine, lysine, and asparagine residues to form a single active site. The structure of Tdp1 confirms that the protein has many similarities to the members of the phospholipase D (PLD) superfamily and indicates a similar catalytic mechanism. The structure also suggests how the unusual protein-DNA substrate binds and provides insights about the nature of the substrate in vivo

The relative contribution of road bicycle components on vibration induced to the cyclist

Improving comfort in road bicycle design is a paramount concern for cyclists, who are affected by the vibrations caused by constant contact with the road surface. The cycling community has deployed many efforts in the attempt to understand and improve bicycle comfort. However, these attempts have been focused on specific components such as the fork, frame and wheels without knowing their relative influence on vibration induced to the bicyclist (VIB). The objective of this paper is to assess the relative contribution of bicycle components on the VIB at the cyclist’s hands and buttocks. A factorial design test comparing the VIB in acceleration, force and power of different bicycle components has already shown that the handlebar and fork are the preponderant components for the VIB measured at the cyclist’s hands. At the buttocks, the preponderant components are the wheels and frame.The authors gratefully acknowledge financial support from the National Science and Engineering Council of Canada (NSERC) and the participation of Cervelo and Vroomen-White Design

Road bike comfort: on the measurement of vibrations induced to cyclist

With ride quality being one of the most sought-after characteristics of a road bicycle by customers as well as by bicycle manufacturers, the vibrational behaviour of the bicycle/cyclist system has grown into an active field in sport engineering research in recent years. When assessing bicycle transmissibility and ride comfort, it is important to control test conditions to obtain repeatable load and acceleration measurements at the cyclist’s contact points with the bicycle. Surprisingly, however, this consideration has not yet been specifically addressed in the literature. The aim of this paper is a first effort to investigate the effect of a selected set of test conditions on the measurement of vibration induced to the cyclist by a road bicycle. Our results showed that all the test conditions selected had a significant effect on the level of vibration induced to the cyclist.The authors gratefully acknowledge financial support from the National Science and Engineering Council of Canada (NSERC) and the participation of Cervélo and Vroomen-White Design

Distinct requirements for the Rad32(Mre¹¹) nuclease and Ctp1(CtIP) in the removal of covalently bound topoisomerase I and II from DNA

For a cancer cell to resist treatment with drugs that trap topoisomerases covalently on the DNA, the topoisomerase must be removed. In this study, we provide evidence that the Schizosaccharomyces pombe Rad32Mre11 nuclease activity is involved in the removal of both Top2 from 5′ DNA ends as well as Top1 from 3′ ends in vivo. A ctp1CtIP deletion is defective for Top2 removal but overproficient for Top1 removal, suggesting that Ctp1CtIP plays distinct roles in removing topoisomerases from 5′ and 3′ DNA ends. Analysis of separation of function mutants suggests that MRN-dependent topoisomerase removal contributes significantly to resistance against topoisomerase-trapping drugs. This study has important implications for our understanding of the role of the MRN complex and CtIP in resistance of cells to a clinically important group of anticancer drugs

Methodological criteria for the assessment of moderators in systematic reviews of randomised controlled trials : a consensus study

Background: Current methodological guidelines provide advice about the assessment of sub-group analysis within RCTs, but do not specify explicit criteria for assessment. Our objective was to provide researchers with a set of criteria that will facilitate the grading of evidence for moderators, in systematic reviews. Method: We developed a set of criteria from methodological manuscripts (n = 18) using snowballing technique, and electronic database searches. Criteria were reviewed by an international Delphi panel (n = 21), comprising authors who have published methodological papers in this area, and researchers who have been active in the study of sub-group analysis in RCTs. We used the Research ANd Development/University of California Los Angeles appropriateness method to assess consensus on the quantitative data. Free responses were coded for consensus and disagreement. In a subsequent round additional criteria were extracted from the Cochrane Reviewers’ Handbook, and the process was repeated. Results: The recommendations are that meta-analysts report both confirmatory and exploratory findings for subgroups analysis. Confirmatory findings must only come from studies in which a specific theory/evidence based apriori statement is made. Exploratory findings may be used to inform future/subsequent trials. However, for inclusion in the meta-analysis of moderators, the following additional criteria should be applied to each study: Baseline factors should be measured prior to randomisation, measurement of baseline factors should be of adequate reliability and validity, and a specific test of the interaction between baseline factors and interventions must be presented. Conclusions: There is consensus from a group of 21 international experts that methodological criteria to assess moderators within systematic reviews of RCTs is both timely and necessary. The consensus from the experts resulted in five criteria divided into two groups when synthesising evidence: confirmatory findings to support hypotheses about moderators and exploratory findings to inform future research. These recommendations are discussed in reference to previous recommendations for evaluating and reporting moderator studies

Effects of DNA and protein size on substrate cleavage by human tyrosyl-DNA phosphodiesterase 1

Tyrosyl-DNA phosphodiesterase 1 (TDP1) catalyzes the hydrolysis of phosphodiester linkages between a DNA 3′ phosphate and a tyrosine residue as well as a variety of other DNA 3′ substituents, and has been implicated in the repair of covalent complexes involving eukaryotic type IB topoisomerases. To better understand the substrate features that are recognized by TDP1, the size of either the DNA or protein component of the substrate was varied. Competition experiments and gel shift analyses comparing a series of substrates with DNA lengths increasing from 6 to 28 nucleotides indicated that, contrary to predictions based on the crystal structure of the protein, the apparent affinity for the substrate increased as the DNA length was increased over the entire range tested. It has previously been found that a substrate containing the full-length native form of human topoisomerase I protein is not cleaved by TDP1. Protein-oligonucleotide complexes containing either a 53 or 108 amino acid long topoisomerase I-derived peptide were efficiently cleaved by TDP1, but like the full length protein, a substrate containing a 333 amino acid topoisomerase I fragment was resistant to cleavage. Consistent with these results, evidence is presented that processing by the proteasome is required for TDP1 cleavage in vivo

BLM and RMI1 alleviate RPA inhibition of topoIIIα decatenase activity

RPA is a single-stranded DNA binding protein that physically associates with the BLM complex. RPA stimulates BLM helicase activity as well as the double Holliday junction dissolution activity of the BLM-topoisomerase IIIα complex. We investigated the effect of RPA on the ssDNA decatenase activity of topoisomerase IIIα. We found that RPA and other ssDNA binding proteins inhibit decatenation by topoisomerase IIIα. Complex formation between BLM, TopoIIIα, and RMI1 ablates inhibition of decatenation by ssDNA binding proteins. Together, these data indicate that inhibition by RPA does not involve species-specific interactions between RPA and BLM-TopoIIIα-RMI1, which contrasts with RPA modulation of double Holliday junction dissolution. We propose that topoisomerase IIIα and RPA compete to bind to single-stranded regions of catenanes. Interactions with BLM and RMI1 enhance toposiomerase IIIα activity, promoting decatenation in the presence of RPA

STK295900, a Dual Inhibitor of Topoisomerase 1 and 2, Induces G<inf>2</inf> Arrest in the Absence of DNA Damage

STK295900, a small synthetic molecule belonging to a class of symmetric bibenzimidazoles, exhibits antiproliferative activity against various human cancer cell lines from different origins. Examining the effect of STK295900 in HeLa cells indicates that it induces G2 phase arrest without invoking DNA damage. Further analysis shows that STK295900 inhibits DNA relaxation that is mediated by topoisomerase 1 (Top 1) and topoisomerase 2 (Top 2) in vitro. In addition, STK295900 also exhibits protective effect against DNA damage induced by camptothecin. However, STK295900 does not affect etoposide-induced DNA damage. Moreover, STK295900 preferentially exerts cytotoxic effect on cancer cell lines while camptothecin, etoposide, and Hoechst 33342 affected both cancer and normal cells. Therefore, STK295900 has a potential to be developed as an anticancer chemotherapeutic agent. © 2013 Kim et al