WorldWideScience

Sample records for single-strand orthodontic wires

  1. [Mechanics analysis of fracture of orthodontic wires].

    Science.gov (United States)

    Wang, Yeping; Sun, Xiaoye; Zhang, Longqi

    2003-03-01

    Fracture problem of orthodontic wires was discussed in this paper. The calculation formulae of bending stress and tensile stress were obtained. All main factors that affect bending stress and tensile stress of orthodontic wires were analyzed and discussed. It was concluded that the main causes of fracture of orthodontic wires were fatigue and static disruption. Some improving proposals for preventing fracture of orthodontic wires were put forward.

  2. Right wire in orthodontics: a review

    OpenAIRE

    Ali, Hashim

    2015-01-01

    Quality of orthodontic wire such as stiffness, hardness, resiliency, elasticity and working range are important determinants of the effectivenes of tooth movement. Commonly used types of orthodontic arch wire:1) stainless steel(ss) wire, 2) conventional nickel- titanium (NiTi)alloy wire,3) improved super elastic NiTi- alloy wire( also called low hysteresis(LH)wire), and titanium molybdenum alloy(TMA) wire.

  3. Initial arch wires for tooth alignment during orthodontic treatment with fixed appliances.

    Science.gov (United States)

    Jian, Fan; Lai, Wenli; Furness, Susan; McIntyre, Grant T; Millett, Declan T; Hickman, Joy; Wang, Yan

    2013-04-30

    potentially confounding factor (such as bracket type, slot size, ligation method, extraction of teeth) which is likely to have influenced the outcome and was not controlled in the trial. None of the trials reported the important adverse outcome of root resorption.Three groups of comparisons were made.(1) Multistrand stainless steel initial arch wires compared to superelastic nickel titanium (NiTi) initial arch wires. There were four trials in this group, with different comparisons and outcomes reported at different times. No meta-analysis was possible. There is insufficient evidence from these trials to determine whether or not there is a difference in either rate of alignment or pain between stainless steel and NiTi initial arch wires.(2) Conventional (stabilised) NiTi initial arch wires compared to superelastic NiTi initial arch wires. There were two trials in this group, one reporting the outcome of alignment over 6 months and the other reporting pain over 1 week. There is insufficient evidence from these trials to determine whether or not there is any difference between conventional (stabilised) and superelastic NiTi initial arch wires with regard to either alignment or pain.(3) Single-strand superelastic NiTi initial arch wires compared to other NiTi (coaxial, copper NiTi (CuNiTi) or thermoelastic) initial arch wires. The three trials in this comparison each compared a different product against single-strand superelastic NiTi. There is very weak unreliable evidence, based on one very small study (n = 24) at high risk of bias, that coaxial superelastic NiTi may produce greater tooth movement over 12 weeks, but no information on associated pain or root resorption. This result should be interpreted with caution until further research evidence is available. There is insufficient evidence to determine whether or not there is a difference between either thermoelastic or CuNiTi and superelastic NiTi initial arch wires. There is no reliable evidence from the trials included in

  4. Load-Deflection and Friction Properties of PEEK Wires as Alternative Orthodontic Wires.

    Science.gov (United States)

    Tada, Yoshifumi; Hayakawa, Tohru; Nakamura, Yoshiki

    2017-08-09

    Polyetheretherketone (PEEK) is now attracting attention as an alternative to metal alloys in the dental field. In the present study, we evaluated the load-deflection characteristics of PEEK wires in addition to their frictional properties. Three types of PEEK wires are used: two sizes of rectangular shape, 0.016 × 0.022 in² and 0.019 × 0.025 in² (19-25PEEK), and rounded shape, diameter 0.016 in (16PEEK). As a control, Ni-Ti orthodontic wire, diameter 0.016 in, was used. The three-point bending properties were evaluated in a modified three-point bending system for orthodontics. The static friction between the orthodontic wire and the bracket was also measured. The load-deflection curves were similar among Ni-Ti and PEEK wires, except for 16PEEK with slot-lid ligation. The bending force of 19-25PEEK wire was comparable with that of Ni-Ti wire. 19-25PEEK showed the highest load at the deflection of 1500 μm ( p 0.05). No significant difference was seen in static friction between all three PEEK wires and Ni-Ti wire ( p > 0.05). It is suggested that 19-25PEEK will be applicable for orthodontic treatment with the use of slot-lid ligation.

  5. Finite element modeling of superelastic nickel-titanium orthodontic wires.

    Science.gov (United States)

    Naceur, Ines Ben; Charfi, Amin; Bouraoui, Tarak; Elleuch, Khaled

    2014-11-28

    Thanks to its good corrosion resistance and biocompatibility, superelastic Ni–Ti wire alloys have been successfully used in orthodontic treatment. Therefore, it is important to quantify and evaluate the level of orthodontic force applied to the bracket and teeth in order to achieve tooth movement. In this study, three dimensional finite element models with a Gibbs-potential-based-formulation and thermodynamic principles were used. The aim was to evaluate the influence of possible intraoral temperature differences on the forces exerted by NiTi orthodontic arch wires with different cross sectional shapes and sizes. The prediction made by this phenomenological model, for superelastic tensile and bending tests, shows good agreement with the experimental data. A bending test is simulated to study the force variation of an orthodontic NiTi arch wire when it loaded up to the deflection of 3 mm, for this task one half of the arch wire and the 3 adjacent brackets were modeled. The results showed that the stress required for the martensite transformation increases with the increase of cross-sectional dimensions and temperature. Associated with this increase in stress, the plateau of this transformation becomes steeper. In addition, the area of the mechanical hysteresis, measured as the difference between the forces of the upper and lower plateau, increases.

  6. Mechanical characterisation of orthodontic superelastic Ni-Ti wires

    Energy Technology Data Exchange (ETDEWEB)

    Arrigoni, M.; Pietrabissa, R. [Politecnico di Milano, Milano (Italy). Lab. of Biological Structure Mechanics; Auricchio, F.; Petrini, L. [Politecnico di Milano, Milano (Italy). Lab. of Biological Structure Mechanics; Pavia Univ. (Italy). Dept. of Structural Mechanics; Cacciafesta, V. [Politecnico di Milano, Milano (Italy). Lab. of Biological Structure Mechanics; Pavia Univ. (Italy). Dept. of Orthodontia

    2001-11-01

    Nowadays, the orthodontic treatment is improving thanks to the introduction of Ni-Ti super-elastic alloy wires in the ordinary therapy. Indeed, laboratory tests performed in the last decade have shown that Ni-Ti superelastic wires are able to satisfy the ideal requirements for fixed arch-wire appliance: high flexibility, minimal distortion or plastic deformation, light constant force production over a wide range of displacements. On the other hand, many orthodontic companies produce Ni-Ti arch-wires, without giving detailed specifications on their superelastic characteristics. To improve the knowledge on real properties for these products, an experimental campaign on different commercial arch-wires has been started at the Laboratory of Biological Structure Mechanics (LABS) at the Politecnico di Milano (Italy). This work presents the first step of the research, concerning the comparison between the behaviour of four types of wires (two produced by ORMCO and two produced by 3M/Unitek) under monotonic and cyclic isothermal tensile tests. The results show significant differences between the products in terms of elastic modulus, stress values of the loading-unloading plateau, hysteresis amplitude, spring-back capacity, shape recovery capability, strain rate effect and fatigue behaviour. (orig.)

  7. Initial arch wires for alignment of crooked teeth with fixed orthodontic braces.

    LENUS (Irish Health Repository)

    Wang, Yan

    2010-01-01

    The initial arch wire is the first arch wire to be inserted into the fixed appliance at the beginning of orthodontic treatment and is used mainly for correcting crowding and rotations of teeth. With a number of orthodontic arch wires available for initial tooth alignment, it is important to understand which wire is most efficient, as well as which wires cause the least amount of root resorption and pain during the initial aligning stage of treatment.

  8. Biofilm formation on stainless steel and gold wires for bonded retainers in vitro and in vivo and their susceptibility to oral antimicrobials.

    Science.gov (United States)

    Jongsma, Marije A; Pelser, Floris D H; van der Mei, Henny C; Atema-Smit, Jelly; van de Belt-Gritter, Betsy; Busscher, Henk J; Ren, Yijin

    2013-05-01

    Bonded retainers are used in orthodontics to maintain treatment result. Retention wires are prone to biofilm formation and cause gingival recession, bleeding on probing and increased pocket depths near bonded retainers. In this study, we compare in vitro and in vivo biofilm formation on different wires used for bonded retainers and the susceptibility of in vitro biofilms to oral antimicrobials. Orthodontic wires were exposed to saliva, and in vitro biofilm formation was evaluated using plate counting and live/dead staining, together with effects of exposure to toothpaste slurry alone or followed by antimicrobial mouthrinse application. Wires were also placed intra-orally for 72 h in human volunteers and undisturbed biofilm formation was compared by plate counting and live/dead staining, as well as by denaturing gradient gel electrophoresis for compositional differences in biofilms. Single-strand wires attracted only slightly less biofilm in vitro than multi-strand wires. Biofilms on stainless steel single-strand wires however, were much more susceptible to antimicrobials from toothpaste slurries and mouthrinses than on single-strand gold wires and biofilms on multi-strand wires. Also, in vivo significantly less biofilm was found on single-strand than on multi-strand wires. Microbial composition of biofilms was more dependent on the volunteer involved than on wire type. Biofilms on single-strand stainless steel wires attract less biofilm in vitro and are more susceptible to antimicrobials than on multi-strand wires. Also in vivo, single-strand wires attract less biofilm than multi-strand ones. Use of single-strand wires is preferred over multi-strand wires, not because they attract less biofilm, but because biofilms on single-strand wires are not protected against antimicrobials as in crevices and niches as on multi-strand wires.

  9. Phase transformation changes in thermocycled nickel-titanium orthodontic wires.

    Science.gov (United States)

    Berzins, David W; Roberts, Howard W

    2010-07-01

    In the oral environment, orthodontic wires will be subject to thermal fluctuations. The purpose of this study was to investigate the effect of thermocycling on nickel-titanium (NiTi) wire phase transformations. Straight segments from single 27 and 35 degrees C copper NiTi (Ormco), Sentalloy (GAC), and Nitinol Heat Activated (3M Unitek) archwires were sectioned into 5mm segments (n=20). A control group consisted of five randomly selected non-thermocycled segments. The remaining segments were thermocycled between 5 and 55 degrees C with five randomly selected segments analyzed with differential scanning calorimetry (DSC; -100150 degrees C at 10 degrees C/min) after 1000, 5000, and 10,000 cycles. Thermal peaks were evaluated with results analyzed via ANOVA (alpha=0.05). Nitinol HA and Sentalloy did not demonstrate qualitative or quantitative phase transformation behavior differences. Significant differences were observed in some of the copper NiTi transformation temperatures, as well as the heating enthalpy with the 27 degrees C copper NiTi wires (p<0.05). Qualitatively, with increased thermocycling the extent of R-phase in the heating peaks decreased in the 35 degrees C copper NiTi, and an austenite to martensite peak shoulder developed during cooling in the 27 degrees C copper NiTi. Repeated temperature fluctuations may contribute to qualitative and quantitative phase transformation changes in some NiTi wires. Copyright 2010 Academy of Dental Materials. All rights reserved.

  10. Microstructure of NiTi orthodontic wires observations using transmission electron microscopy

    OpenAIRE

    Ferčec, J.; Jenko, D.; Buchmeister, B.; Rojko, F.; Budič, B.; Kosec, B.; Rudolf, R.

    2014-01-01

    This work presents the results of the microstructure observation of six different types of NiTi orthodontic wires by using Transmission Electron Microscopy (TEM). Within these analyses the chemical compositions of each wire were observed in different places by applying the EDS detector. Namely, the chemical composition in the orthodontic wires is very important because it shows the dependence between the phase temperatures and mechanical properties. Microstructure observations showed that ort...

  11. Evaluation of Effects of Sterilization on Mechanical Properties of Orthodontic Wires

    Directory of Open Access Journals (Sweden)

    Sridhar Kannan

    2012-01-01

    Results: Dry heat sterilization, autoclave, 2% glutaraldehyde solution had no effect on ultimate tensile strength, 0.1% yield strength, modulus of elasticity and percentage elongation of stainless steel and elgiloy wires. Tensile strength and yield strength of Nitinol and b-titanium wires together with percentage elongation of b-titanium wires significantly increased following dry heat sterilization and autoclave. No detrimental effects on properties of wires were observed. These sterilization procedures could be safely recommended for sterilization of orthodontic wires.

  12. Surface characterization of nickel titanium orthodontic arch wires

    Science.gov (United States)

    Krishnan, Manu; Seema, Saraswathy; Tiwari, Brijesh; Sharma, Himanshu S.; Londhe, Sanjay; Arora, Vimal

    2015-01-01

    Background Surface roughness of nickel titanium orthodontic arch wires poses several clinical challenges. Surface modification with aesthetic/metallic/non metallic materials is therefore a recent innovation, with clinical efficacy yet to be comprehensively evaluated. Methods One conventional and five types of surface modified nickel titanium arch wires were surface characterized with scanning electron microscopy, energy dispersive analysis, Raman spectroscopy, Atomic force microscopy and 3D profilometry. Root mean square roughness values were analyzed by one way analysis of variance and post hoc Duncan's multiple range tests. Results Study groups demonstrated considerable reduction in roughness values from conventional in a material specific pattern: Group I; conventional (578.56 nm) > Group V; Teflon (365.33 nm) > Group III; nitride (301.51 nm) > Group VI (i); rhodium (290.64 nm) > Group VI (ii); silver (252.22 nm) > Group IV; titanium (229.51 nm) > Group II; resin (158.60 nm). It also showed the defects with aesthetic (resin/Teflon) and nitride surfaces and smooth topography achieved with metals; titanium/silver/rhodium. Conclusions Resin, Teflon, titanium, silver, rhodium and nitrides were effective in decreasing surface roughness of nickel titanium arch wires albeit; certain flaws. Findings have clinical implications, considering their potential in lessening biofilm adhesion, reducing friction, improving corrosion resistance and preventing nickel leach and allergic reactions. PMID:26843749

  13. Autoclaving and clinical recycling: Effects on mechanical properties of orthodontic wires

    Directory of Open Access Journals (Sweden)

    M Oshagh

    2012-01-01

    Conclusion: Although recycle wires were softer than those of control group, relatively small differences and also various properties of available wires have obscured the clinical predictability of their application. There is seemingly no problem in terms of mechanical properties to recycle orthodontic wires.

  14. Adhesive Properties of Bonded Orthodontic Retainers to Enamel : Stainless Steel Wire vs Fiber-reinforced Composites

    NARCIS (Netherlands)

    Foek, Dave Lie Sam; Krebs, Eliza; Sandham, John; Ozcan, Mutlu

    2009-01-01

    Purpose: The objectives of this study were to compare the bond strength of a stainless steel orthodontic wire vs various fiber-reinforced composites (FRC) used as orthodontic retainers on enamel, analyze the failure types after debonding, and investigate the influence of different application

  15. Torque resistance of different stainless steel wires commonly used for fixed retainers in orthodontics

    DEFF Research Database (Denmark)

    Arnold, Dario; Dalstra, Michel; Verna, Carlalberta

    2016-01-01

    Objective: Movements of teeth splinted by fixed retention wires after orthodontic treatment have been observed. The aetiological factors for these movements are unknown. The aim of this in vitro study was to compare the resistance to torque of different stainless steel wires commonly used for fixed...... retainers in orthodontics. Materials and Methods: Torquing moments acting on a retainer wire were measured in a mechanical force testing system by applying buccal crown torque to an upper lateral incisor in both a 3-teeth and in a 2-teeth setup. Seven stainless steel wires with different shape, type (plain...... or a braided 0.016 × 0.022-inch stainless steel wire. A tooth attached by a retainer wire to only one neighbouring tooth is less resistant to torque than a tooth connected to two neighbouring teeth. Annealing a retainer wire with a flame reduces the stiffness of the wire markedly and can lead to a non...

  16. Mechanical properties and aesthetics of FRP orthodontic wire fabricated by hot drawing.

    Science.gov (United States)

    Imai, T; Watari, F; Yamagata, S; Kobayashi, M; Nagayama, K; Toyoizumi, Y; Nakamura, S

    1998-12-01

    The FRP wires 0.5 mm in diameter with a multiple fiber structure were fabricated by drawing the fiber polymer complex at 250 degrees C for an esthetic, transparent orthodontic wire. Biocompatible CaO-P2O5-SiO2-Al2O3 (CPSA) glass fibers of 8-20 microm in diameter were oriented unidirectionally in the longitudinal direction in PMMA matrix. The mechanical properties were investigated by 3-point flexural test. The FRP wire showed sufficient strength and a very good elastic recovery after deformation. Young's modulus and the flexural load at deflection 1 mm were nearly independent of the fiber diameter and linearly increased with the fiber fraction. The dependence on fiber fraction obeys well the rule of mixture. This FRP wire could cover the range of strength corresponding to the conventional metal orthodontic wires from Ni-Ti used in the initial stage of orthodontic treatments to Co-Cr used in the final stage by changing the volume ratio of glass fibers with the same external diameter. The estheticity in external appearance was excellent. Thus the new FRP wire can satisfy both mechanical properties necessary for an orthodontic wire and enough estheticity, which was not possible for the conventional metal wire.

  17. Effect of Sodium Fluoride Mouthwash on the Frictional Resistance of Orthodontic Wires

    Directory of Open Access Journals (Sweden)

    Allahyar Geramy

    2017-12-01

    Full Text Available Objectives: The friction between the brackets and orthodontic wire during sliding mechanics inflicts difficulties such as decreasing the applied force and tooth movement and also the loss of anchorage. Therefore, many studies have focused on the factors that affect the friction. The purpose of this study was to assess the effect of 0.05% sodium fluoride mouthwash on the friction between orthodontic brackets and wire.Materials and Methods: Four types of orthodontic wires including rectangular standard stainless steel (SS, titanium molybdenum alloy (TMA, nickel-titanium (NiTi and copper-nickel-titanium (Cu-NiTi were selected. In each group, half of the samples were immersed in 0.05% sodium fluoride mouthwash and the others were immersed in artificial saliva for 10 hours. An elastomeric ligature was used for ligating the wires to brackets. The frictional test was performed in a universal testing machine at the speed of 10 mm/minute. Two-way ANOVA was used for statistical analysis of the friction rate.Results: The friction rate was significantly higher after immersion in 0.05% sodium fluoride mouthwash in comparison with artificial saliva (P=0.00. Cu-NiTi wire showed the highest friction value followed by TMA, NiTi and SS wires.  Conclusions: According to the results of the current study, 0.05% sodium fluoride mouthwash increased the frictional characteristics of all the evaluated orthodontic wires.

  18. Galvanic coupling of steel and gold alloy lingual brackets with orthodontic wires.

    Science.gov (United States)

    Polychronis, Georgios; Al Jabbari, Youssef S; Eliades, Theodore; Zinelis, Spiros

    2018-03-06

    The aim of this research was to assess galvanic behavior of lingual orthodontic brackets coupled with representative types of orthodontic wires. Three types of lingual brackets: Incognito (INC), In-Ovation L (IOV), and STb (STB) were combined with a stainless steel (SS) and a nickel-titanium (NiTi) orthodontic archwire. All materials were initially investigated by scanning electron microscopy / x-ray energy dispersive spectroscopy (SEM/EDX) while wires were also tested by x-ray diffraction spectroscopy (XRD). All bracket-wire combinations were immersed in acidic 0.1M NaCl 0.1M lactic acid and neutral NaF 0.3% (wt) electrolyte, and the potential differences were continuously recorded for 48 hours. The SEM/EDX analysis revealed that INC is a single-unit bracket made of a high gold (Au) alloy while IOV and STB are two-piece appliances in which the base and wing are made of SS alloys. The SS wire demonstrated austenite and martensite iron phase, while NiTi wire illustrated an intense austenite crystallographic structure with limited martensite. All bracket wire combinations showed potential differences below the threshold of galvanic corrosion (200 mV) except for INC and STB coupled with NiTi wire in NaF media. The electrochemical results indicate that all brackets tested demonstrated galvanic compatibility with SS wire, but fluoride treatment should be used cautiously with NiTi wires coupled with Au and SS brackets.

  19. Mechanical properties of orthodontic wires on ceramic brackets associated with low friction ligatures

    Directory of Open Access Journals (Sweden)

    Fernando KOIKE

    2017-03-01

    Full Text Available Abstract Introduction Few studies investigated the mechanical properties of orthodontic wires on ceramic brackets associated the ligatures. Objective This study aimed to compare the load-deflection of orthodontic wires with round section of 0.016” made of stainless steel (SS, nickel-titanium (NiTi and glass fiber-reinforced polymer composite (GFRPC. Material and method Sixty specimens obtained from 10 sectioned pre-contoured arches (TP Orthodontics, were divided into 3 groups of 20 according to each type of material (1 esthetic-type wire and 2 not esthetic and length of 50 mm. The methodology consisted of a 3-point bending test using esthetic ceramic brackets (INVU, TP Orthodontics, Edgewise, 0.022”x 0.025” as points of support. The tensile tests were performed on a mechanical test machine, at a speed of 10 mm/min, deflection of 1 mm, 2 mm and 3 mm. Friedman’s Non Parametric Multiple comparisons test was used (P<0.05. Result The nickel-titanium wire presented smaller load/ deflection compared with stainless steel. GFRPC wires had lower strength values among all groups evaluated (P<.05. The steel wire showed permanent deformation after 3 mm deflection, NiTi wire demonstrated memory effect and the esthetic type had fractures with loss of strength. Conclusion It can be concluded that steel wires have high strength values, requiring the incorporation of loops and folds to reduce the load / deflection. NiTi and GFRPC wires produced low levels of force, however the esthetic wire was shown to fracture and break.

  20. The effect of surface treatment and clinical use on friction in NiTi orthodontic wires.

    Science.gov (United States)

    Wichelhaus, Andrea; Geserick, Marc; Hibst, Raimund; Sander, Franz G

    2005-10-01

    Since the low friction of NiTi wires allows a rapid and efficient orthodontic tooth movement, the aim of this research was to investigate the friction and surface roughness of different commercially available superelastic NiTi wires before and after clinical use. The surface of all of the wires had been pre-treated by the manufacturer. Forty superelastic wires (Titanol Low Force, Titanol Low Force River Finish Gold, Neo Sentalloy, Neo Sentalloy Ionguard) of diameter 0.016 x 0.022 in. were tested. The friction for each type of NiTi archwire ligated into a commercial stainless steel bracket was determined with a universal testing machine. Having ligated the wire into the bracket, it could then be moved forward and backwards along a fixed archwire whilst a torquing moment was applied. The surface roughness was investigated using a profilometric measuring device on defined areas of the wire. Statistical data analysis was conducted by means of the Wilcoxon test. The results showed that initially, the surface treated wires demonstrated significantly (p < 0.01) less friction than the non-treated wires. The surface roughness showed no significant difference between the treated and the non-treated surfaces of the wires. All 40 wires however showed a significant increase in friction and surface roughness during clinical use. Whilst the Titanol Low Force River Finish Gold (Forestadent, Pforzheim, Germany) wires showed the least friction of all the samples and consequently should be more conservative on anchorage, the increase in friction of all the surface treated wires during orthodontic treatment almost cancels out this initial effect on friction. It is therefore recommended that surface treated NiTi orthodontic archwires should only be used once.

  1. Bacterial colonization on coated and uncoated orthodontic wires: A prospective clinical trial

    OpenAIRE

    Seyed Hamid Raji; Hasan Shojaei; Parinaz Saeidi Ghorani; Elahe Rafiei

    2014-01-01

    Background: The advantages of coated orthodontic wires such as esthetic and their effects on reduced friction, corrosion and allergic reaction and the significant consequences of plaque accumulation on oral health encouraged us to assess bacterial colonization on these wires. Materials and Methods: A total of 18 (9 upper and 9 lower) epoxy resin coated 16 × 22 nickel-titanium wires (Spectra, GAC, USA) and 18 (9 upper and 9 lower) non-coated 16 × 22 nickel-titanium wires (Sentalloy, GAC, U...

  2. Microstructure of NiTi orthodontic wires observations using transmission electron microscopy

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    J. Ferčec

    2014-10-01

    Full Text Available This work presents the results of the microstructure observation of six different types of NiTi orthodontic wires by using Transmission Electron Microscopy (TEM. Within these analyses the chemical compositions of each wire were observed in different places by applying the EDS detector. Namely, the chemical composition in the orthodontic wires is very important because it shows the dependence between the phase temperatures and mechanical properties. Microstructure observations showed that orthodontic wires consist of nano-sized grains containing precipitates of Ti2Ni and/or TiC. The first precipitated Ti2Ni are rich in Ti, while the precipitated TiC is rich in C. Further investigation showed that there was a difference in average grain size in the NiTi matrix. The sizes of grains in orthodontic wires are in the range from approximately 50 to 160 nm and the sizes of precipitate are in the range from 0,3 μm to 5 μm.

  3. Bacterial colonization on coated and uncoated orthodontic wires: A prospective clinical trial.

    Science.gov (United States)

    Raji, Seyed Hamid; Shojaei, Hasan; Ghorani, Parinaz Saeidi; Rafiei, Elahe

    2014-11-01

    The advantages of coated orthodontic wires such as esthetic and their effects on reduced friction, corrosion and allergic reaction and the significant consequences of plaque accumulation on oral health encouraged us to assess bacterial colonization on these wires. A total of 18 (9 upper and 9 lower) epoxy resin coated 16 × 22 nickel-titanium wires (Spectra, GAC, USA) and 18 (9 upper and 9 lower) non-coated 16 × 22 nickel-titanium wires (Sentalloy, GAC, USA) with isolated packages were selected and sterilized before application. The samples were divided randomly between upper and lower arches in 18 patients and hence that every patient received one coated and one uncoated wire at the same time. Samples were removed and cut in equal lengths after 3 weeks and placed in phosphate buffered saline buffer. After separation of bacteria in trypsin and ethylenediaminetetraacetic acid solution, the diluted solution was cultured in blood agar and bacterial colony forming units were counted. Finally, the data was analyzed using the paired t-test and the significance was set at 0.05. Mean of bacterial colonization on uncoated wires was more than that of coated wires (P < 0.001). Bacterial plaque accumulation on epoxy resin coated nickel-titanium orthodontic wires is significantly lower than uncoated nickel-titanium wires.

  4. Deflection and Flexural Strength Effects on the Roughness of Aesthetic-Coated Orthodontic Wires

    OpenAIRE

    Albuquerque, Cibele Gonçalves de; Correr, Américo Bortolazzo; Venezian, Giovana Cherubini; Santamaria Jr, Milton; Tubel, Carlos Alberto; Vedovello, Silvia Amélia Scudeler

    2017-01-01

    Abstract The aim was to evaluate the flexural strength and the effects of deflection on the surface roughness of esthetic orthodontic wires. The sample consisted of 70 archwire 0.014-inch: polytetrafluorethylene (PTFE)-coated Nickel-Titanium (Niti) archwires (Titanol Cosmetic-TC, Flexy Super Elastic Esthetic-FSE, esthetic Nickel Titanium Wire-ANT); epoxy resin-coated Niti archwires (Spectra-S, Niticosmetic-TEC); gold and rhodium coated Niti (Sentalloy-STC) and a control group (superelastic Ni...

  5. Initial arch wires for alignment of crooked teeth with fixed orthodontic braces.

    Science.gov (United States)

    Wang, Yan; Jian, Fan; Lai, Wenli; Zhao, Zhihe; Yang, Zhi; Liao, Zhengyu; Shi, Zongdao; Wu, Taixiang; Millett, Declan T; McIntyre, Grant T; Hickman, Joy

    2010-04-14

    The initial arch wire is the first arch wire to be inserted into the fixed appliance at the beginning of orthodontic treatment and is used mainly for correcting crowding and rotations of teeth. With a number of orthodontic arch wires available for initial tooth alignment, it is important to understand which wire is most efficient, as well as which wires cause the least amount of root resorption and pain during the initial aligning stage of treatment. To identify and assess the evidence for the effects of initial arch wires for alignment of teeth with fixed orthodontic braces in relation to alignment speed, root resorption and pain intensity. We searched the following electronic databases: the Cochrane Oral Health Group's Trials Register (30th November 2009), CENTRAL (The Cochrane Library 2009, Issue 4), MEDLINE (1950 to 30th November 2009) and EMBASE (1980 to 30th November 2009). Reference lists of articles were also searched. There was no restriction with regard to publication status or language of publication. We contacted all authors of included studies to identify additional studies. Randomised controlled trials (RCTs) of initial arch wires to align crooked teeth with fixed orthodontic braces were selected. Only studies involving patients with upper and/or lower full arch fixed orthodontic appliances were included. Two review authors were responsible for study selection, validity assessment and data extraction. All disagreements were resolved by discussion amongst the review team. Corresponding authors of included studies were contacted to obtain missing information. Seven RCTs, with 517 participants, provided data for this review. Among them, five trials investigated the speed of initial tooth alignment comparing: 0.016 inch ion-implanted A-NiTi wire versus 0.016 inch A-NiTi versus 0.0175 multistrand stainless steel wire; 0.016x0.022 inch medium force active M-NiTi wire versus 0.016x0.022 inch graded force active M-NiTi wire versus 0.0155 inch multistrand

  6. Deflection and Flexural Strength Effects on the Roughness of Aesthetic-Coated Orthodontic Wires.

    Science.gov (United States)

    Albuquerque, Cibele Gonçalves de; Correr, Américo Bortolazzo; Venezian, Giovana Cherubini; Santamaria, Milton; Tubel, Carlos Alberto; Vedovello, Silvia Amélia Scudeler

    2017-01-01

    The aim was to evaluate the flexural strength and the effects of deflection on the surface roughness of esthetic orthodontic wires. The sample consisted of 70 archwire 0.014-inch: polytetrafluorethylene (PTFE)-coated Nickel-Titanium (Niti) archwires (Titanol Cosmetic-TC, Flexy Super Elastic Esthetic-FSE, esthetic Nickel Titanium Wire-ANT); epoxy resin-coated Niti archwires (Spectra-S, Niticosmetic-TEC); gold and rhodium coated Niti (Sentalloy-STC) and a control group (superelastic Niti (Nitinol-NS). The initial roughness was evaluated with a rugosimeter. After that, the wires were submitted to flexural test in an universal testing machine. Each wire was deflected up to 2 mm at a speed of 1 mm/min. After flexural test, the roughness of the wires was evaluted on the same surface as that used for the initial evaluation. The data of roughness and flexural strength were analyzed by one-way ANOVA and Tukey's test (a=0.05). Student t-test compared roughness before and after deflection (a =0.05). The roughness of S and ANT (epoxy resin and PTFE-coated wires, respectively), before and after deflection, was significantly higher than the other groups (p<0.05). Wire deflection significantly increased the roughness of the wires S and STC (p<0.05). The flexural strength of groups FSE and NS (PTFE and uncoated) was higher compared with that of the other groups (p<0.05). We concluded that the roughness and flexural strength of the orthodontic wires does not depend on the type of the esthetic coating, but it is influenced by the method of application of this coating. The deflection can increase the roughness of the esthetic orthodontic wires.

  7. Nickel release from orthodontic retention wires: the action of mechanical loading and pH

    NARCIS (Netherlands)

    Milheiro, A.; Kleverlaan, C.; Muris, J.; Feilzer, A.; Pallav, P.

    2012-01-01

    Nickel (Ni) is a potent sensitizer and may induce innate and adaptive immune responses. Ni is an important component of orthodontic appliances (8-50 wt%). Due to chemical and mechanical factors in the oral environment, Ni is released from these appliances. Retention wires are in situ for a long

  8. Accuracy of four different digital intraoral scanners: effects of the presence of orthodontic brackets and wire.

    Science.gov (United States)

    Jung, Yoo-Ran; Park, Ji-Man; Chun, Youn-Sic; Lee, Kkot-Nim; Kim, Minji

    The objective of this study was to compare the accuracy of four different digital intraoral scanners and the effects of buccal brackets and orthodontic wire. For this study, three sets of models (Control model, BKT model with buccal bracket, and WBKT model with buccal bracket and orthodontic wire) were scanned using four different types of intraoral scanners: E4D dentist, iTero, Trios, and Zfx IntraScan. The mesiodistal width of the teeth, intercanine width, and intermolar width measured by four scanners were compared. Three-dimensional (3D) images of the brackets were taken using the four scanners. Data were analyzed with one-way ANOVA, independent t test, and post-hoc Tukey test at a significance level of P brackets and orthodontic wire. Comparison of 3D bracket images scanned by the four scanners showed differences in image distortion among the scanners. Bracket characteristics did not affect the 3D bracket images. The four intraoral scanners used in this study differed in accuracy. However, the results acquired by iTero and Trios were more reliable. Effects of buccal brackets and orthodontic wire on the 3D images taken by intraoral scanners were not clinically significant.

  9. Development and evaluation of a reinforced polymeric biomaterial for use as an orthodontic wire

    Science.gov (United States)

    Zufall, Scott William

    Composite archwires have the potential to provide esthetic and functional improvements over conventional wires. As part of an ongoing effort to bring these materials into general use, composite wires were fabricated using a photo-pultrusion manufacturing technique, and subsequently coated with a 10 mum layer of poly(chloro-p-xylylene). Coated and uncoated composites were subjected to several different evaluations to assess their ability to perform the functions of an orthodontic archwire. An investigation of the viscoelastic behavior of uncoated composite wires was conducted at a physiological temperature of 37°C using a bend stress relaxation test. Over 90 day testing periods, energy losses increased with decreasing reinforcement levels from to 8% of the initial wire stress. Final viscous losses were 1% for all reinforcement levels. Relaxed elastic moduli for the composite wires were comparable to the reported elastic moduli of conventional orthodontic wires that are typically used for initial and intermediate alignment procedures. Frictional characteristics were evaluated in passive and active configurations for uncoated composite wires against three contemporary orthodontic brackets. Kinetic coefficients of friction were the same for all wire-bracket combinations tested and were slightly lower than the reported coefficients of other initial and intermediate alignment wires. Wear patterns on the wires, which were largely caused by sharp leading edges of the bracket slots, were characteristic of plowing and cutting wear behaviors. This wear caused glass fibers to be released from the surface of the wires, presenting a potential irritant. Coated composite wires were subjected to the same frictional analysis as the uncoated wires. A mathematical model of the archwire-bracket system was derived using engineering mechanics, and used to define a coefficient of binding. The coating increased the frictional coefficients of the wires by 72%, yet the binding coefficient

  10. Torque resistance of different stainless steel wires commonly used for fixed retainers in orthodontics.

    Science.gov (United States)

    Arnold, Dario T; Dalstra, Michel; Verna, Carlalberta

    2016-06-01

    Movements of teeth splinted by fixed retention wires after orthodontic treatment have been observed. The aetiological factors for these movements are unknown. The aim of this in vitro study was to compare the resistance to torque of different stainless steel wires commonly used for fixed retainers in orthodontics. Torquing moments acting on a retainer wire were measured in a mechanical force testing system by applying buccal crown torque to an upper lateral incisor in both a 3-teeth and in a 2-teeth setup. Seven stainless steel wires with different shape, type (plain, braided, coaxial, or chain) and dimensions were selected for this study. For a torquing angle of 16.2° in the 3-teeth setup torsion moments can vary between 390 cNmm and 3299 cNmm depending on the retainer wire. For the 2-teeth setup the torsion moments are much smaller. Exposure to the flame of a butane-gas torch for 10 seconds to anneal the wire reduces the stiffness of the retainer wire. Clinicians must select wires for fixed retainers very carefully since the difference in resistance to torque is large. A high level of torque control can be achieved with a plain 0.016 × 0.016-inch or a braided 0.016 × 0.022-inch stainless steel wire. A tooth attached by a retainer wire to only one neighbouring tooth is less resistant to torque than a tooth connected to two neighbouring teeth. Annealing a retainer wire with a flame reduces the stiffness of the wire markedly and can lead to a non-uniform and non-reproducible effect.

  11. Release of nickel and chromium ions from orthodontic wires following the use of teeth whitening mouthwashes

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    AmirHossein Mirhashemi

    2018-02-01

    Full Text Available Abstract Background Corrosion resistance is an important requirement for orthodontic appliances. Nickel and chromium may be released from orthodontic wires and can cause allergic reactions and cytotoxicity when patients use various mouthwashes to whiten their teeth. Our study aimed to assess the release of nickel and chromium ions from nickel titanium (NiTi and stainless steel (SS orthodontic wires following the use of four common mouthwashes available on the market. Methods This in vitro, experimental study was conducted on 120 orthodontic appliances for one maxillary quadrant including five brackets, one band and half of the required length of SS, and NiTi wires. The samples were immersed in Oral B, Oral B 3D White Luxe, Listerine, and Listerine Advance White for 1, 6, 24, and 168 h. The samples immersed in distilled water served as the control group. Atomic absorption spectroscopy served to quantify the amount of released ions. Results Nickel ions were released from both wires at all time-points; the highest amount was in Listerine and the lowest in Oral B mouthwashes. The remaining two solutions were in-between this range. The process of release of chromium from the SS wire was the same as that of nickel. However, the release trend in NiTi wires was not uniform. Conclusions Listerine caused the highest release of ions. Listerine Advance White, Oral B 3D White Luxe, and distilled water were the same in terms of ion release. Oral B showed the lowest amount of ion release.

  12. Characterization and coating stability evaluation of nickel-titanium orthodontic esthetic wires: an in vivo study

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    Nina ARGALJI

    2017-08-01

    Full Text Available Abstract The objective of this study was to compare coating dimensions and surface characteristics of two different esthetic covered nickel-titanium orthodontic rectangular archwires, as-received from the manufacturer and after oral exposure. The study was designed for comparative purposes. Both archwires, as-received from the manufacturer, were observed using a stereomicroscope to measure coating thickness and inner metallic dimensions. The wires were also exposed to oral environment in 11 orthodontic active patients for 21 days. After removing the samples, stereomicroscopy images were captured, coating loss was measured and its percentage was calculated. Three segments of each wire (one as-received and two after oral exposure were observed using scanning electron microscopy for a qualitative analysis of the labial surface of the wires. The Lilliefors test and independent t-test were applied to verify normality of data and statistical differences between wires, respectively. The significance level adopted was 0.05. The results showed that the differences between the wires while comparing inner height and thickness were statistically significant (p < 0.0001. In average, the most recently launched wire presented a coating thickness twice that of the control wire, which was also a statistically significant difference. The coating loss percentage was also statistically different (p = 0.0346 when the latest launched wire (13.27% was compared to the control (29.63%. In conclusion, the coating of the most recent wire was thicker and more uniform, whereas the control had a thinner coating on the edges. After oral exposure, both tested wires presented coating loss, but the most recently launched wire exhibited better results.

  13. Characterization and coating stability evaluation of nickel-titanium orthodontic esthetic wires: an in vivo study.

    Science.gov (United States)

    Argalji, Nina; Silva, Eduardo Moreira da; Cury-Saramago, Adriana; Mattos, Claudia Trindade

    2017-08-21

    The objective of this study was to compare coating dimensions and surface characteristics of two different esthetic covered nickel-titanium orthodontic rectangular archwires, as-received from the manufacturer and after oral exposure. The study was designed for comparative purposes. Both archwires, as-received from the manufacturer, were observed using a stereomicroscope to measure coating thickness and inner metallic dimensions. The wires were also exposed to oral environment in 11 orthodontic active patients for 21 days. After removing the samples, stereomicroscopy images were captured, coating loss was measured and its percentage was calculated. Three segments of each wire (one as-received and two after oral exposure) were observed using scanning electron microscopy for a qualitative analysis of the labial surface of the wires. The Lilliefors test and independent t-test were applied to verify normality of data and statistical differences between wires, respectively. The significance level adopted was 0.05. The results showed that the differences between the wires while comparing inner height and thickness were statistically significant (p wire presented a coating thickness twice that of the control wire, which was also a statistically significant difference. The coating loss percentage was also statistically different (p = 0.0346) when the latest launched wire (13.27%) was compared to the control (29.63%). In conclusion, the coating of the most recent wire was thicker and more uniform, whereas the control had a thinner coating on the edges. After oral exposure, both tested wires presented coating loss, but the most recently launched wire exhibited better results.

  14. The Effect of Annealing on the Elastic Modulus of Orthodontic Wires

    Science.gov (United States)

    Higginbottom, Kyle

    Introduction: Nickel Titanium orthodontic wires are currently used in orthodontic treatment due to their heat activated properties and their delivery of constant force. The objective of this study was to determine the effect of annealing on the elastic modulus of Nickel Titanium, Stainless Steel and Beta-titanium (TMA) wires. Different points along the wire were tested in order to determine how far from the annealed ends the elastic modulus of the wires was affected. Methods: Eighty (80) orthodontic wires consisting of 4 equal groups (SS/TMA/Classic NitinolRTM/Super Elastic NitinolRTM) were used as the specimens for this study. All wires were measured and marked at 5mm measurements, and cut into 33.00mm sections. The wires were heated with a butane torch until the first 13.00mm of the wires were red hot. Load deflection tests using an InstronRTM universal testing machine were run at 5mm distances from the end of the wire that had been annealed. The change in elastic modulus was then determined. Results: There was a significant difference (F = 533.001, p = 0.0005) in the change in elastic modulus for the four distances. There was also a significant difference (F = 57.571, p = 0.0005) in the change in elastic modulus for the four wire types. There was a significant interaction (F = 19.601, p = 0.005) between wire type and distance, however this interaction negated the differences between the wires. Conclusion: 1) There are significant differences in the changes in elastic modulus between the areas of the wires within the annealed section and those areas 5mm and 10mm away from the annealed section. The change in elastic modulus within the annealed section was significantly greater at 8 mm than it was at 13mm, and this was significantly greater than 18mm and 23mm (5mm and 10mm beyond the annealed section). However, there was no statistical difference in the change in elastic modulus between 5mm and 10mm away from the annealed section (18mm and 23mm respectively). 2

  15. Martensitic transformation of austenitic stainless steel orthodontic wires during intraoral exposure.

    Science.gov (United States)

    Izquierdo, Paula P; de Biasi, Ronaldo S; Elias, Carlos N; Nojima, Lincoln I

    2010-12-01

    Our purpose was to study the mechanical properties and phase transformations of orthodontic wires submitted to in-vivo exposure in the mouth for different periods of time. Stainless steel wires were tied to fixed orthodontic appliances of 30 patients from the orthodontics clinic of Universidade Federal do Rio de Janeiro School of Dentistry in Brazil. According to the duration of the clinical treatment, the patients were divided into 3 groups. After in-vivo exposure, the samples were studied by mechanical testing (torsion) and ferromagnetic resonance. Statistical analyses were carried out to evaluate the correlation between time of exposure, mechanical properties, and austenite-to-martensite transformation among the groups. The results were compared with as-received control samples. The torque values increased as time in the mouth increased. The increase in torque resistance showed high correlations with time of exposure (P = 0.005) and austenite-martensite phase transformation. The resistance of stainless steel orthodontic wires increases as the time in the mouth increases; this effect is attributed to the austenite-to-martensite transformation. Copyright © 2010 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.

  16. Ion Release and Galvanic Corrosion of Different Orthodontic Brackets and Wires in Artificial Saliva.

    Science.gov (United States)

    Tahmasbi, Soodeh; Sheikh, Tahereh; Hemmati, Yasamin B

    2017-03-01

    To investigate the galvanic corrosion of brackets manufactured by four different companies coupled with stainless steel (SS) or nickel-titanium (NiTi) wires in an artificial saliva solution. A total of 24 mandibular central incisor Roth brackets of four different manufacturers (American Orthodontics, Dentaurum, Shinye, ORJ) were used in this experimental study. These brackets were immersed in artificial saliva along with SS or NiTi orthodontic wires (0.016'', round) for 28 days. The electric potential difference of each bracket/ wire coupled with a saturated calomel reference electrode was measured via a voltmeter and recorded constantly. Corrosion rate (CR) was calculated, and release of ions was measured with an atomic absorption spectrometer. Stereomicroscope was used to evaluate all samples. Then, samples with corrosion were further assessed by scanning electron microscope and energy-dispersive X-ray spectroscopy. Two-way analysis of variance was used to analyze data. Among ions evaluated, release of nickel ions from Shinye brackets was significantly higher than that of other brackets. The mean potential difference was significantly lower in specimens containing a couple of Shinye brackets and SS wire compared with other specimens. No significant difference was observed in the mean CR of various groups (p > 0.05). Microscopic evaluation showed corrosion in two samples only: Shinye bracket coupled with SS wire and American Orthodontics bracket coupled with NiTi wire. Shinye brackets coupled with SS wire showed more susceptibility to galvanic corrosion. There were no significant differences among specimens in terms of the CR or released ions except the release of Ni ions, which was higher in Shinye brackets.

  17. Effects of a diamond-like carbon coating on the frictional properties of orthodontic wires.

    Science.gov (United States)

    Muguruma, Takeshi; Iijima, Masahiro; Brantley, William A; Mizoguchi, Itaru

    2011-01-01

    To test the hypothesis that a diamond-like carbon coating does not affect the frictional properties of orthodontic wires. Two types of wires (nickel-titanium and stainless steel) were used, and diamond-like carbon (DLC) films were deposited on the wires. Three types of brackets, a conventional stainless steel bracket and two self-ligating brackets, were used for measuring static friction. DLC layers were observed by three-dimensional scanning electron microscopy (3D-SEM), and the surface roughness was measured. Hardness and elastic modulus were obtained by nanoindentation testing. Frictional forces and surface roughness were compared by the Kruskal-Wallis and Mann-Whitney U-tests. The hardness and elastic modulus of the wires were compared using Student's t-test. When angulation was increased, the DLC-coated wires showed significantly less frictional force than the as-received wires, except for some wire/bracket combinations. Thin DLC layers were observed on the wire surfaces by SEM. As-received and DLC-coated wires had similar surface morphologies, and the DLC-coating process did not affect the surface roughness. The hardness of the surface layer of the DLC-coated wires was much higher than for the as-received wires. The elastic modulus of the surface layer of the DLC-coated stainless steel wire was less than that of the as-received stainless steel wire, whereas similar values were found for the nickel-titanium wires. The hypothesis is rejected. A DLC-coating process does reduce the frictional force.

  18. Force systems in the initial phase of orthodontic treatment -- a comparison of different leveling arch wires.

    Science.gov (United States)

    Fuck, Lars-Michael; Drescher, Dieter

    2006-01-01

    The determination of orthodontically-effective forces and moments places great demands on the technical equipment. Many patients report severe pain after fixed appliance insertion. Since it is assumed that pain from orthodontic appliances is associated with the force and moment levels applied to the teeth and since the occurrence of root resorption is a common therapeutic side effect, it would seem important to know the actual magnitudes of the components of the active orthodontic force systems. The aim of the present study was therefore to measure initial force systems produced by different leveling arch-wires in a complete multi-bracket appliance and to assess whether force and moment levels can be regarded as biologically acceptable or not. The actual bracket position in 42 patients was transferred onto a measurement model. Forces and moments produced by a super-elastic nickel-titanium (NiTi) archwire, a 6-strand stainless steel archwire, and a 7-strand super-elastic NiTi archwire were determined experimentally on different teeth. Average forces and moments produced by the super-elastic NiTi arch wires were found to be the highest. In spite if their larger diameter, the stranded arch wires' average force and moment levels were lower, especially that of the stranded super-elastic archwire. Nevertheless, maximum force levels sometimes exceeded recommended values in the literature and must be considered as too high. The measured arch wires' initial force systems differed significantly depending on the type of archwire and its material structure. Stranded arch wires produced lower force and moment levels, and we recommend their use in the initial phase of orthodontic treatment.

  19. Relationship between friction force and orthodontic force at the leveling stage using a coated wire

    Directory of Open Access Journals (Sweden)

    Masaki MURAYAMA

    2013-12-01

    Full Text Available The relationship between orthodontic force and friction produced from an archwire and brackets affects the sliding of the wire in the leveling stage. Objective: The purpose of this study was to evaluate the relationship between force and friction in a small esthetic nickel-titanium (Ni-Ti wire. Material and Methods: Five esthetic wires (three coated and two plated and two small, plain Ni-Ti wires (0.012 and 0.014 inches were used. We performed a three-point bending test according to ISO 15841 and the drawing test with a dental arch model designed with upper linguoversion of the lateral incisor in the arch (displacements of 0.5, 1.0, 2.0 and 3.0 mm, and evaluated the relationship between them. Results: Unloading bending forces of all wires at displacements of less than 1.0 mm were larger than friction forces, but all friction forces at displacements exceeding 2.0 mm were larger than unloading bending forces. The arch likely expands when displacement from the proximal brackets exceeds 1.0 mm. The friction force of a martensite 0.014-inch Ni-Ti wire was significantly greater than those of the other esthetic and austenitic wires. Conclusions: A wire with the smallest possible friction force should be used in cases with more than 1.0 mm displacement.

  20. Relationship between friction force and orthodontic force at the leveling stage using a coated wire.

    Science.gov (United States)

    Murayama, Masaki; Namura, Yasuhiro; Tamura, Takahiko; Iwai, Hiroaki; Shimizu, Noriyoshi

    2013-01-01

    The relationship between orthodontic force and friction produced from an archwire and brackets affects the sliding of the wire in the leveling stage. The purpose of this study was to evaluate the relationship between force and friction in a small esthetic nickel-titanium (Ni-Ti) wire. Five esthetic wires (three coated and two plated) and two small, plain Ni-Ti wires (0.012 and 0.014 inches) were used. We performed a three-point bending test according to ISO 15841 and the drawing test with a dental arch model designed with upper linguoversion of the lateral incisor in the arch (displacements of 0.5, 1.0, 2.0 and 3.0 mm), and evaluated the relationship between them. Unloading bending forces of all wires at displacements of less than 1.0 mm were larger than friction forces, but all friction forces at displacements exceeding 2.0 mm were larger than unloading bending forces. The arch likely expands when displacement from the proximal brackets exceeds 1.0 mm. The friction force of a martensite 0.014-inch Ni-Ti wire was significantly greater than those of the other esthetic and austenitic wires. A wire with the smallest possible friction force should be used in cases with more than 1.0 mm displacement.

  1. The Mechanical Behavior Variation of Nickel - Titanium Orthodontic Wires in Different Fluoride Mouthwash

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    Ilnaz haj Hemati

    2016-12-01

    Full Text Available In this study, an orthodontic arch wire of Ni-Ti alloy made by Germany in four different mouthwashes contain fluoride was investigated in mechanical behavior. This research focused on the mechanical behavior changes in different mouthwashes. The Mouthwashes were chosen from most popular one in Iran including Oral-B, Gum, and Behsa Mouthwash. The wires after 3 month in touch with mouthwashes were studied and the result was presented. The result show that the lowest power to reshape the wire, is for the wire in Crest case (3. The maximum power assigns to wire in Behsa case (1. Also, the shortest treatment period is for wire in Behsa mouthwash case (1 with 0.087 Nm strain energy and the maximum duration of therapy is related to case (3 Crest mouthwash with 0.039 Nm. Moreover, the wires in Oral-b (0.095 Nm has the lowest and Gum mouthwash (0.140 Nm has the highest energy intake. The energy absorbed by the wires in Behsa and Crest is in the middle. Finally, it is proposed that patient could used all mouthwash but based on their condition, one product will be useful.

  2. Corrosion resistance of titanium-containing dental orthodontic wires in fluoride-containing artificial saliva

    International Nuclear Information System (INIS)

    Lee, T.-H.; Wang, C.-C.; Huang, T.-K.; Chen, L.-K.; Chou, M.-Y.; Huang, H.-H.

    2009-01-01

    This study was to investigate the corrosion resistance of different Ti-containing dental orthodontic wires (including Ni-Ti, Ni-Ti-Cu, Ti-Mo-Zr-Sn, and Ti-Nb alloys) in acidic fluoride-containing artificial saliva using cyclic potentiodynamic polarization curve measurements. Different NaF concentrations (0%, 0.2%, and 0.5%), simulating the fluoride contents in commercial toothpastes, were added to the artificial saliva. Surface characterization was analyzed using X-ray photoelectron spectrometry. Cyclic potentiodynamic polarization curves showed that the presence of fluoride ions, especially 0.5% NaF, was detrimental to the protective ability of the TiO 2 -based film on the Ti-containing wires. This might lead to a decrease in the corrosion resistance of the tested alloys, i.e. an increase in the corrosion rate and anodic current density and a decrease in the passive film breakdown potential. Among the tested Ti-containing wires, the Ni-Ti and Ni-Ti-Cu wires containing mainly TiO 2 on surface film were more susceptible to fluoride-enhanced corrosion, while the Ti-Mo-Zr-Sn and Ti-Nb wires containing MoO 3 /ZrO 2 /SnO and Nb 2 O 5 , respectively, along with TiO 2 on surface film were pitting corrosion resistant and showed a lower susceptibility to fluoride-enhanced corrosion. The difference in corrosion resistance of the tested commercial Ti-containing dental orthodontic wires was significantly dependent on the passive film characteristics on wires' surface.

  3. Corrosion resistance of titanium-containing dental orthodontic wires in fluoride-containing artificial saliva

    Energy Technology Data Exchange (ETDEWEB)

    Lee, T.-H. [Institute of Medicine, Chung Shan Medical University, Taichung 402, Taiwan (China); Department of Dentistry, Chung Shan Medical University Hospital, Taichung 402, Taiwan (China); Wang, C.-C. [Department of Dental Laboratory Technology, Min-Hwei College of Health Care Management, Tainan County 736, Taiwan (China); Huang, T.-K. [College of Dental Medicine, Kaohsiung Medical University, Kaohsiung 807, Taiwan (China); Chen, L.-K. [Department of Dentistry, Taipei City Hospital, Taipei 115, Taiwan (China); Chou, M.-Y. [Institute of Medicine, Chung Shan Medical University, Taichung 402, Taiwan (China); Department of Dentistry, Chung Shan Medical University Hospital, Taichung 402, Taiwan (China); Huang, H.-H., E-mail: hhhuang@ym.edu.t [Department of Dentistry, Taipei City Hospital, Taipei 115, Taiwan (China); Department of Dentistry, National Yang-Ming University, Taipei 112, Taiwan (China); Institute of Oral Biology, National Yang-Ming University, Taipei 112, Taiwan (China)

    2009-11-20

    This study was to investigate the corrosion resistance of different Ti-containing dental orthodontic wires (including Ni-Ti, Ni-Ti-Cu, Ti-Mo-Zr-Sn, and Ti-Nb alloys) in acidic fluoride-containing artificial saliva using cyclic potentiodynamic polarization curve measurements. Different NaF concentrations (0%, 0.2%, and 0.5%), simulating the fluoride contents in commercial toothpastes, were added to the artificial saliva. Surface characterization was analyzed using X-ray photoelectron spectrometry. Cyclic potentiodynamic polarization curves showed that the presence of fluoride ions, especially 0.5% NaF, was detrimental to the protective ability of the TiO{sub 2}-based film on the Ti-containing wires. This might lead to a decrease in the corrosion resistance of the tested alloys, i.e. an increase in the corrosion rate and anodic current density and a decrease in the passive film breakdown potential. Among the tested Ti-containing wires, the Ni-Ti and Ni-Ti-Cu wires containing mainly TiO{sub 2} on surface film were more susceptible to fluoride-enhanced corrosion, while the Ti-Mo-Zr-Sn and Ti-Nb wires containing MoO{sub 3}/ZrO{sub 2}/SnO and Nb{sub 2}O{sub 5}, respectively, along with TiO{sub 2} on surface film were pitting corrosion resistant and showed a lower susceptibility to fluoride-enhanced corrosion. The difference in corrosion resistance of the tested commercial Ti-containing dental orthodontic wires was significantly dependent on the passive film characteristics on wires' surface.

  4. Effect of thermocycling on nickel release from orthodontic arch wires: an in vitro study.

    Science.gov (United States)

    Sheibaninia, Ahmad

    2014-12-01

    The amount of daily intake of metals from orthodontic appliances over time is a matter of great concern. Nickel results in one of the most common metal-induced allergic contact dermatitis in humans; it produces more allergic reactions than all the other metals combined together. The purpose of this study was to evaluate the effects of thermocycling on the nickel release from orthodontic arch wires stored in artificial saliva with different pH values. Forty new wire pieces were selected. Each wire piece was placed in a special capillary Pyrex tube filled with artificial saliva, which was sealed and immersed in deionized water at 37 °C. The samples were divided into four groups of ten. Group I received no treatment; group II was subjected to thermocycling. The pH of storage in groups III and IV was reduced to 4.5, and group IV was subjected to thermocycling. Thermocycling was carried out between 5 and 55 °C for 500 cycles. The release of nickel ions was statistically analyzed by two-way ANOVA for the effects of two variables: pH and thermocycling. The interaction between pH and thermocycling was found to be statistically significant (F = 12.127, P = 0.001). Two-way ANOVA showed that different storage media or pH and thermocycling had a significant effect on the nickel release (F = 52.812, P nickel from orthodontic wires, while thermocycling is clearly the dominant factor.

  5. Influence of stress and phase on corrosion of a superelastic nickel-titanium orthodontic wire.

    Science.gov (United States)

    Segal, Nadav; Hell, Jess; Berzins, David W

    2009-06-01

    The purpose of this investigation was to study the effect of stress and phase transformation on the corrosion properties of a superelastic nickel-titanium orthodontic wire. The phase transformation profiles of superelastic nickel-titanium (Sentalloy, GAC International, Bohemia, NY) and beta-titanium (TMA, Ormco, Orange, Calif) archwires were analyzed by using differential scanning calorimetry. The force/deflection behavior of the wires at 37 degrees C was measured in a 3-point bending test per modified American Dental Association specification no. 32. Electrochemical testing consisted of monitoring the open circuit potential (OCP) for 2 hours followed by polarization resistance and cyclic polarization tests on archwire segments engaged in a 5-bracket simulation apparatus with bend deflections of 0.75, 1.5, or 3 mm in artificial saliva at 37 degrees C. Nondeflected segments were also tested. Sentalloy was additionally examined for bending and corrosion at 5 degrees C, where it exists as martensite and is devoid of stress-induced phase transformation. OCP at 2 hours and corrosion current density (i(corr)) were analyzed by using ANOVA and Tukey tests (alpha = .05) (n = 10 per deflection). Significant differences (P Sentalloy wires at 5 degrees C, but not for Sentalloy at 37 degrees C. Significant differences (P Sentalloy (37 degrees C) peaked at 0.75 mm deflection before the wire's stress-induced phase transformation point and then decreased with further deflection and transformation. The i(corr) values for TMA and Sentalloy at 5 degrees C, both of which do not undergo phase transformation with deformation, continuously increased from 0 to 1.5 mm deflection before decreasing at the 3.0-mm deflection. Stress increased the corrosion rate in nickel-titanium and beta-titanium orthodontic wires. Alterations in stress/strain associated with phase transformation in superelastic nickel-titanium might alter the corrosion rate in ways different from wires not undergoing phase

  6. Mechanical properties of orthodontic wires covered with a polyether ether ketone tube.

    Science.gov (United States)

    Shirakawa, Nobukazu; Iwata, Toshio; Miyake, Shinjiro; Otuka, Takero; Koizumi, So; Kawata, Toshitugu

    2018-03-21

    To evaluate the esthetics and frictional force of an orthodontic wire passed through a newly designed tube made of a polyether ether ketone (PEEK) resin. Two types of standard PEEK tubes were prepared at 0.5 × 0.6ф and 0.8 × 0.9ф, and different archwires were passed through the tubes. Color values were determined according to brightness and hues. Friction was assessed with different bracket-wire combinations, and surface roughness was determined by stereomicroscopy before and after the application of friction. The PEEK tube showed a color difference that was almost identical to that of coated wires conventionally used in clinical practice, indicating a sufficient esthetic property. The result of the friction test showed that the frictional force was greatly reduced by passing the archwire through the PEEK tube in almost all of the archwires tested. Use of the new PEEK tube demonstrated a good combination of esthetic and functional properties for use in orthodontic appliances.

  7. Influence of autoclave sterilization on the surface parameters and mechanical properties of six orthodontic wires.

    Science.gov (United States)

    Pernier, C; Grosgogeat, B; Ponsonnet, L; Benay, G; Lissac, M

    2005-02-01

    Orthodontic wires are frequently packaged in individual sealed bags in order to avoid cross-contamination. The instructions on the wrapper generally advise autoclave sterilization of the package and its contents if additional protection is desired. However, sterilization can modify the surface parameters and the mechanical properties of many types of material. The aim of this research was to determine the influence of one of the most widely used sterilization processes, autoclaving (18 minutes at 134 degrees C, as recommended by the French Ministry of Health), on the surface parameters and mechanical properties of six wires currently used in orthodontics (one stainless steel alloy: Tru-Chrome RMO; two nickel-titanium shape memory alloys: Neo Sentalloy and Neo Sentalloy with Ionguard GAC; and three titanium-molybdenum alloys: TMA(R) and Low Friction TMA Ormco and Resolve GAC). The alloys were analysed on receipt and after sterilization, using surface structure observation techniques, including optical, scanning electron and atomic force microscopy and profilometry. The mechanical properties were assessed by three-point bending tests. The results showed that autoclave sterilization had no adverse effects on the surface parameters or on the selected mechanical properties. This supports the possibility for practitioners to systematically sterilize wires before placing them in the oral environment.

  8. Assessment of the hardness of different orthodontic wires and brackets produced by metal injection molding and conventional methods.

    Science.gov (United States)

    Alavi, Shiva; Kachuie, Marzie

    2017-01-01

    This study was conducted to assess the hardness of orthodontic brackets produced by metal injection molding (MIM) and conventional methods and different orthodontic wires (stainless steel, nickel-titanium [Ni-Ti], and beta-titanium alloys) for better clinical results. A total of 15 specimens from each brand of orthodontic brackets and wires were examined. The brackets (Elite Opti-Mim which is produced by MIM process and Ultratrimm which is produced by conventional brazing method) and the wires (stainless steel, Ni-Ti, and beta-titanium) were embedded in epoxy resin, followed by grinding, polishing, and coating. Then, X-ray energy dispersive spectroscopy (EDS) microanalysis was applied to assess their elemental composition. The same specimen surfaces were repolished and used for Vickers microhardness assessment. Hardness was statistically analyzed with Kruskal-Wallis test, followed by Mann-Whitney test at the 0.05 level of significance. The X-ray EDS analysis revealed different ferrous or co-based alloys in each bracket. The maximum mean hardness values of the wires were achieved for stainless steel (SS) (529.85 Vickers hardness [VHN]) versus the minimum values for beta-titanium (334.65 VHN). Among the brackets, Elite Opti-Mim exhibited significantly higher VHN values (262.66 VHN) compared to Ultratrimm (206.59 VHN). VHN values of wire alloys were significantly higher than those of the brackets. MIM orthodontic brackets exhibited hardness values much lower than those of SS orthodontic archwires and were more compatible with NiTi and beta-titanium archwires. A wide range of microhardness values has been reported for conventional orthodontic brackets and it should be considered that the manufacturing method might be only one of the factors affecting the mechanical properties of orthodontic brackets including hardness.

  9. Interlot variations of transition temperature range and force delivery in copper-nickel-titanium orthodontic wires.

    Science.gov (United States)

    Pompei-Reynolds, Renée C; Kanavakis, Georgios

    2014-08-01

    The manufacturing process for copper-nickel-titanium archwires is technique sensitive. The primary aim of this investigation was to examine the interlot consistency of the mechanical properties of copper-nickel-titanium wires from 2 manufacturers. Wires of 2 sizes (0.016 and 0.016 × 0.022 in) and 3 advertised austenite finish temperatures (27°C, 35°C, and 40°C) from 2 manufacturers were tested for transition temperature ranges and force delivery using differential scanning calorimetry and the 3-point bend test, respectively. Variations of these properties were analyzed for statistical significance by calculating the F statistic for equality of variances for transition temperature and force delivery in each group of wires. All statistical analyses were performed at the 0.05 level of significance. Statistically significant interlot variations in austenite finish were found for the 0.016 in/27°C (P = 0.041) and 0.016 × 0.022 in/35°C (P = 0.048) wire categories, and in austenite start for the 0.016 × 0.022 in/35°C wire category (P = 0.01). In addition, significant variations in force delivery were found between the 2 manufacturers for the 0.016 in/27°C (P = 0.002), 0.016 in/35.0°C (P = 0.049), and 0.016 × 0.022 in/35°C (P = 0.031) wires. Orthodontic wires of the same material, dimension, and manufacturer but from different production lots do not always have similar mechanical properties. Clinicians should be aware that copper-nickel-titanium wires might not always deliver the expected force, even when they come from the same manufacturer, because of interlot variations in the performance of the material. Copyright © 2014 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.

  10. Stress distribution in delayed replanted teeth splinted with different orthodontic wires: a three-dimensional finite element analysis.

    Science.gov (United States)

    de Souza, Fernando Isquierdo; Poi, Wilson Roberto; da Silva, Vanessa Ferreira; Martini, Ana Paula; Melo, Regis Alexandre da Cunha; Panzarini, Sonia Regina; Rocha, Eduardo Passos

    2015-06-01

    The aim was to evaluate the biomechanical behavior of the supporting bony structures of replanted teeth and the periodontal ligament (PDL) of adjacent teeth when orthodontic wires with different mechanical properties are applied, with three-dimensional finite element analysis. Based on tomographic and microtomographic data, a three-dimensional model of the anterior maxilla with the corresponding teeth (tooth 13-tooth 23) was generated to simulate avulsion and replantation of the tooth 21. The teeth were splinted with orthodontic wire (Ø 0.8 mm) and composite resin. The elastic modulus of the three orthodontic wires used, that is, steel wire (FA), titanium-molybdenum wire (FTM), and nitinol wire (FN) were 200 GPa, 84 GPa, and 52 GPa, respectively. An oblique load (100 N) was applied at an angle of 45° on the incisal edge of the replanted tooth and was analyzed using Ansys Workbench software. The maximum (σmax) and minimum (σmin) principal stresses generated in the PDL, cortical and alveolar bones, and the modified von Mises (σvM) values for the orthodontic wires were obtained. With regard to the cortical bone and PDL, the highest σmin and σmax values for FTM, FN, and FA were checked. With regard to the alveolar bone, σmax and σmin values were highest for FA, followed by FTM and FN. The σvM values of the orthodontic wires followed the order of rigidity of the alloys, that is, FA > FTM > FN. The biomechanical behavior of the analyzed structures with regard to all the three patterns of flexibility was similar. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. Estimation of changes in nickel and chromium content in nickel-titanium and stainless steel orthodontic wires used during orthodontic treatment: An analytical and scanning electron microscopic study

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    Vandana Kararia

    2015-01-01

    Full Text Available Introduction: The biocompatibility of orthodontic dental alloys has been investigated over the past 20 years, but the results have been inconclusive. The study compares standard 3 M Unitek nickel-titanium (NiTi and stainless steel archwires with locally available JJ orthodontics wires. Scanning electron microscope (SEM study of surface changes and complexometric titration to study compositional change was performed. Materials and Methods: Ten archwires each of group 1-3 M 0.016" NiTi, group 2-JJ 0.016" NiTi, group 3-3 M 0.019" FNx010.025" SS and group 4-JJ SS contributed a 10 mm piece of wire for analysis prior to insertion in the patient and 6 weeks post insertion. SEM images were recorded at Χ2000, Χ4000 and Χ6000 magnification. The same samples were subjected to complexiometric titration using ethylenediaminetetraacetic acid to gauge the actual change in the composition. Observations and Results: The SEM images of all the archwires showed marked changes with deep scratches and grooves and dark pitting corrosion areas post intraoral use. 3M wires showed an uniform criss-cross pattern in as received wires indicating a coating which was absent after intraoral use. There was a significant release of Nickel and Chromium from both group 3 and 4. Group 2 wires released ions significantly more than group 1 (P = 0.0. Conclusion: Extensive and stringent trials are required before certifying any product to be used in Orthodontics.

  12. The effect of ligation on the load deflection characteristics of nickel titanium orthodontic wire.

    Science.gov (United States)

    Kasuya, Shugo; Nagasaka, Satoshi; Hanyuda, Ai; Ishimura, Sadao; Hirashita, Ayao

    2007-12-01

    This study examined the effect of ligation on the load-deflection characteristics of nickel-titanium (NiTi) orthodontic wire. A modified three-point bending system was used for bending the NiTi round wire, which was inserted and ligated in the slots of three brackets, one of which was bonded to each of the three bender rods. Three different ligation methods, stainless steel ligature (SSL), slot lid (SL), and elastomeric ligature (EL), were employed, as well as a control with neither bracket nor ligation (NBL). The tests were repeated five times under each condition. Comparisons were made of load-deflection curve, load at maximum deflection of 2,000 microm, and load at a deflection of 1,500 microm during unloading. Analysis of Variance (ANOVA) and Dunnett's test were conducted to determine method difference (alpha = 0.05). The interaction between deflection and ligation was tested, using repeated-measures ANOVA (alpha = 0.05). The load values of the ligation groups were two to three times greater than the NBL group at a deflection of 1,500 microm during unloading: 4.37 N for EL, 3.90 N for SSL, 3.02 N for SL, and 1.49 N for NBL (P wire may make NiTi wire exhibit a significantly heavier load than that traditionally expected. NiTi wire exhibited the majority of its true superelasticity with SL, whereas EL may act as a restraint on its superelasticity.

  13. Evaluation of force released by deflection of orthodontic wires in conventional and self-ligating brackets.

    Science.gov (United States)

    Higa, Rodrigo Hitoshi; Semenara, Nayara Thiago; Henriques, José Fernando Castanha; Janson, Guilherme; Sathler, Renata; Fernandes, Thais Maria Freire

    2016-01-01

    The aim of the study was to evaluate deflection forces of rectangular orthodontic wires in conventional (MorelliTM), active (In-Ovation RTM) and passive (Damon 3MXTM) self-ligating brackets. Two brands of stainless steel and nickel-titanium (NiTi) wires (MorelliTM and GACTM), in addition to OrmcoTM copper-nickel-titanium wires were used. Specimens were assembled in a clinical simulation device especially designed for this study and tested in an Instron universal testing machine. For the testing procedures, an acrylic structure representative of the maxillary right central incisor was lingually moved in activations of 0 to 1 mm, with readings of the force released by deflection in unloading of 0.5, 0.8 and 1 mm at a constant speed of 2 mm/min. Inter-bracket forces with stainless steel, NiTi and CuNiTi were individually compared by two-way ANOVA, followed by Tukey's tests. Results showed that there were lower forces in conventional brackets, followed by active and passive self-ligating brackets. Within the brands, only for NiTi wires, the MorelliTM brand presented higher forces than GACTM wires. Bracket systems provide different degrees of deflection force, with self-ligating brackets showing the highest forces.

  14. Evaluation of surface roughness of orthodontic wires by means of atomic force microscopy.

    Science.gov (United States)

    D'Antò, Vincenzo; Rongo, Roberto; Ametrano, Gianluca; Spagnuolo, Gianrico; Manzo, Paolo; Martina, Roberto; Paduano, Sergio; Valletta, Rosa

    2012-09-01

    To compare the surface roughness of different orthodontic archwires. Four nickel-titanium wires (Sentalloy(®), Sentalloy(®) High Aesthetic, Titanium Memory ThermaTi Lite(®), and Titanium Memory Esthetic(®)), three β-titanium wires (TMA(®), Colored TMA(®), and Beta Titanium(®)), and one stainless-steel wire (Stainless Steel(®)) were considered for this study. Three samples for each wire were analyzed by atomic force microscopy (AFM). Three-dimensional images were processed using Gwiddion software, and the roughness average (Ra), the root mean square (Rms), and the maximum height (Mh) values of the scanned surface profile were recorded. Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Tukey's post hoc test (P Sentalloy High Aesthetic was the roughest (Ra  =  133.5 ± 10.8; Rms  =  165.8 ± 9.8; Mh  =  949.6 ± 192.1) of the archwires. The surface quality of the wires investigated differed significantly. Ion implantation effectively reduced the roughness of TMA. Moreover, Teflon(®)-coated Titanium Memory Esthetic was less rough than was ion-implanted Sentalloy High Aesthetic.

  15. Hole hopping rates in single strand oligonucleotides

    Energy Technology Data Exchange (ETDEWEB)

    Borrelli, Raffaele [Dipartimento di Scienze Agrarie, Forestali e Alimentari, Università di Torino, Largo Paolo Braccini 2, I-10095 Grugliasco, TO (Italy); Capobianco, Amedeo [Dipartimento di Chimica e Biologia, Università di Salerno, Via Giovanni Paolo II, I-84084 Fisciano, SA (Italy); Peluso, Andrea, E-mail: apeluso@unisa.it [Dipartimento di Chimica e Biologia, Università di Salerno, Via Giovanni Paolo II, I-84084 Fisciano, SA (Italy)

    2014-08-31

    Highlights: • DNA hole transfer rates have been computed. • Delocalized adenine domains significantly affect hole transfer rates in DNA. • Franck–Condon weighted density of state from DFT normal modes. • DNA application in molecular electronics. - Abstract: The rates of hole transfer between guanine and adenine in single strand DNA have been evaluated by using Fermi’s golden rule and Kubo’s generating function approach for the Franck–Condon weighted density of states. The whole sets of the normal modes and vibrational frequencies of the two nucleobases, obtained at DFT/B3LYP level of calculation, have been considered in computations. The results show that in single strand the pyramidalization/planarization mode of the amino groups of both nucleobases plays the major role. At room temperature, the Franck–Condon density of states extends over a wide range of hole site energy difference, 0–1 eV, giving some hints about the design of oligonucleotides of potential technological interest.

  16. Wire Roughness Assessment of 0.016'' × 0.022'' the Technique Lingual Orthodontics.

    Science.gov (United States)

    Facchini, Fátima Mm; Filho, Mario Vedovello; Vedovello, Silvia As; Cotrim, Flávio A; Cotrim-Ferreira, Andrຟa; Tubel, Carlos Am

    2017-04-01

    To evaluate the difference in surface roughness of stainless steel archwires of different commercial brands used in lingual orthodontics. Precontoured arches measuring 0.016'' × 0.022'' were selected of the following brands: Tecnident, Adenta, G&H, Highland Metals Inc., Ormco, Incognito, and Ebraces. Quantitative evaluation of the surface roughness of archwires was performed by means of an atomic force microscope in contact mode. Three surface readouts were taken of each sample, analyzing areas of 20 × 20 μm. Each scan of the samples produced a readout of 512 lines, generating three-dimensional images of the wires. The analysis of variance statistical test was applied to prove significant variables (p > 0.05), with H 0 being rejected and H 1 accepted. The Incognito brand showed the lowest surface roughness. The archwires of brands Adenta, Tecnident, Highland, and Ormco showed similar values among them, and all close to these obtained by the Incognito brand. The archwires of the Ebraces brand showed the highest surface roughness, with values being close to those of the G&H Brand. There was a statistical difference in surface roughness of orthodontic archwires among the brands studied. Companies should pay attention to the quality control of their materials, as these may directly affect the quality of orthodontic treatment.

  17. Metal release profiles of orthodontic bands, brackets, and wires: an in vitro study.

    Science.gov (United States)

    Wendl, B; Wiltsche, H; Lankmayr, E; Winsauer, H; Walter, A; Muchitsch, A; Jakse, N; Wendl, M; Wendl, T

    2017-11-01

    The present study evaluated the temporal release of Co Cr, Mn, and Ni from the components of a typical orthodontic appliance during simulated orthodontic treatment. Several commercially available types of bands, brackets, and wires were exposed to an artificial saliva solution for at least 44 days and the metals released were quantified in regular intervals using inductively coupled plasma quadrupole mass spectrometry (ICP-MS, Elan DRC+, Perkin Elmer, USA). Corrosion products encountered on some products were investigated by a scanning electron microscope equipped with an energy dispersive X-ray microanalyzer (EDX). Bands released the largest quantities of Co, Cr, Mn, and Ni, followed by brackets and wires. Three different temporal metal release profiles were observed: (1) constant, though not necessarily linear release, (2) saturation (metal release stopped after a certain time), and (3) an intermediate release profile that showed signs of saturation without reaching saturation. These temporal metal liberation profiles were found to be strongly dependent on the individual test pieces. The corrosion products which developed on some of the bands after a 6-month immersion in artificial saliva and the different metal release profiles of the investigated bands were traced back to different attachments welded onto the bands. The use of constant release rates will clearly underestimate metal intake by the patient during the first couple of days and overestimate exposure during the remainder of the treatment which is usually several months long. While our data are consistent with heavy metal release by orthodontic materials at levels well below typical dietary intake, we nevertheless recommend the use of titanium brackets and replacement of the band with a tube in cases of severe Ni or Cr allergy.

  18. Evaluation of deflection forces of orthodontic wires with different ligation types

    Directory of Open Access Journals (Sweden)

    José Fernando Castanha HENRIQUES

    2017-07-01

    Full Text Available Abstract The aim of this study was to evaluate deflection forces of orthodontic wires of different alloys engaged into conventional brackets using several ligation types. Stainless steel, conventional superelastic nickel-titanium and thermally activated nickel-titanium archwires tied into conventional brackets by a ring-shaped elastomeric ligature (RSEL, a 8-shaped elastomeric ligature (8SEL and a metal ligature (ML were tested. A clinical simulation device was created especially for this study and forces were measured with an Instron Universal Testing Machine. For the testing procedure, the block representing the maxillary right central incisor was moved 0.5 and 1 mm bucco-lingually at a constant speed of 2 mm/min, and the forces released by the wires were recorded, in accordance with the ISO 15841 guidelines. In general, the RSEL showed lighter forces, while 8SEL and ML showed higher values. At the 0.5 mm deflection, the 8SEL presented the greatest force, but at the 1.0 mm deflection the ML had a statistically similar force. Based on our evaluations, to obtain lighter forces, the thermally activated nickel-titanium wire with the RSEL are recommended, while the steel wire with the 8SEL or the ML are recommended when larger forces are desired. The ML exhibited the highest force increase with increased deflections, compared with the elastomeric ligatures.

  19. Galvanic Corrosion of and Ion Release from Various Orthodontic Brackets and Wires in a Fluoride-containing Mouthwash

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    Soodeh Tahmasbi

    2015-09-01

    Full Text Available Background and aims. This study compared the galvanic corrosion of orthodontic wires and brackets from various man-ufacturers following exposure to a fluoride mouthwash. Materials and methods. This study was conducted on 24 lower central incisor 0.022” Roth brackets of four different commercially available brands (Dentaurum, American Orthodontics, ORJ, Shinye. These brackets along with stainless steel (SS or nickel-titanium (NiTi orthodontic wires (0.016", round were immersed in Oral-B mouthwash containing 0.05% sodium fluoride for 28 days. The electric potential (EP difference of each bracket-wire couple was measured with a Satu-rated Calomel Reference Electrode (Ag/AgCl saturated with KCl via a voltmeter. The ions released in the electrolyte weremeasured with an atomic absorption spectrometer. All the specimens were assessed under a stereomicroscope and speci-mens with corrosion were analyzed with scanning electron microscopy (SEM. Data were analyzed using ANOVA. Results. The copper ions released from specimens with NiTi wire were greater than those of samples containing SS wire. ORJ brackets released more Cu ions than other samples. The Ni ions released from Shinye brackets were significantly more than those of other specimens (P < 0.05. Corrosion rate of brackets coupled with NiTi wires was higher than that of brack-ets coupled with SS wires. Light and electron microscopic observations showed greater corrosion of ORJ brackets. Conclusion. In fluoride mouthwash, Shinye and ORJ brackets exhibited greater corrosion than Dentaurum and American Orthodontics brackets. Stainless steel brackets used with NiTi wires showed greater corrosion and thus caution is recom-mended when using them.

  20. Galvanic Corrosion of and Ion Release from Various Orthodontic Brackets and Wires in a Fluoride-containing Mouthwash.

    Science.gov (United States)

    Tahmasbi, Soodeh; Ghorbani, Mohammad; Masudrad, Mahdis

    2015-01-01

    Background and aims. This study compared the galvanic corrosion of orthodontic wires and brackets from various manufacturers following exposure to a fluoride mouthwash. Materials and methods. This study was conducted on 24 lower central incisor 0.022" Roth brackets of four different commercially available brands (Dentaurum, American Orthodontics, ORJ, Shinye). These brackets along with stainless steel (SS) or nickel-titanium (NiTi) orthodontic wires (0.016", round) were immersed in Oral-B mouthwash containing 0.05% sodium fluoride for 28 days. The electric potential (EP) difference of each bracket-wire couple was measured with a Saturated Calomel Reference Electrode (Ag/AgCl saturated with KCl) via a voltmeter. The ions released in the electrolyte weremeasured with an atomic absorption spectrometer. All the specimens were assessed under a stereomicroscope and specimens with corrosion were analyzed with scanning electron microscopy (SEM). Data were analyzed using ANOVA. Results. The copper ions released from specimens with NiTi wire were greater than those of samples containing SS wire. ORJ brackets released more Cu ions than other samples. The Ni ions released from Shinye brackets were significantly more than those of other specimens (P brackets coupled with NiTi wires was higher than that of brackets coupled with SS wires. Light and electron microscopic observations showed greater corrosion of ORJ brackets. Conclusion. In fluoride mouthwash, Shinye and ORJ brackets exhibited greater corrosion than Dentaurum and American Orthodontics brackets. Stainless steel brackets used with NiTi wires showed greater corrosion and thus caution is recommended when using them.

  1. The effect of ZnO nanoparticle coating on the frictional resistance between orthodontic wires and ceramic brackets

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    Ahmad

    2016-06-01

    Full Text Available Background. Any decrease in friction between orthodontic wire and bracket can accelerate tooth movement in the sliding technique and result in better control of anchorage. This study was carried out to evaluate frictional forces by coating orthodontic wires and porcelain brackets with zinc oxide nanoparticles (ZnO. Methods. In this in vitro study, we evaluated a combination of 120 samples of 0.019×0.025 stainless steel (SS orthodontic wires and 22 mil system edgewise porcelain brackets with and without spherical zinc oxide nanoparticles. Spherical ZnO nanoparticles were deposited on wires and brackets by immersing them in ethanol solution and SEM (scanning electron microscope evaluation confirmed the presence of the ZnO coating. The frictional forces were calculated between the wires and brackets in four groups: group ZZ (coated wire and bracket, group OO (uncoated wire and bracket, group ZO (coated wire and uncoated bracket and group OZ (uncoated wire and coated bracket. Kolmogorov-Smirnov, Mann-Whitney and Kruskal-Wallis tests were used for data analysis. Results. The frictional force in ZZ (3.07±0.4 N was the highest (P <0.05, and OZ (2.18±0.5 N had the lowest amount of friction (P <0.05 among the groups. There was no significant difference in frictional forces between the ZO and OO groups (2.65±0.2 and 2.70±0.2 N, respectively. Conclusion. Coating of porcelain bracket surfaces with ZnO nanoparticles can decrease friction in the sliding technique, and wire coating combined with bracket coating is not recommended due to its effect on friction.

  2. Effect of coating on properties of esthetic orthodontic nickel-titanium wires.

    Science.gov (United States)

    Iijima, Masahiro; Muguruma, Takeshi; Brantley, William; Choe, Han-Cheol; Nakagaki, Susumu; Alapati, Satish B; Mizoguchi, Itaru

    2012-03-01

    To determine the effect of coating on the properties of two esthetic orthodontic nickel-titanium wires. Woowa (polymer coating; Dany Harvest) and BioForce High Aesthetic Archwire (metal coating; Dentsply GAC) with cross-section dimensions of 0.016 × 0.022 inches were selected. Noncoated posterior regions of the anterior-coated Woowa and uncoated Sentalloy were used for comparison. Nominal coating compositions were determined by x-ray fluorescence (JSX-3200, JOEL). Cross-sectioned and external surfaces were observed with a scanning electron microscope (SEM; SSX-550, Shimadzu) and an atomic force microscope (SPM-9500J2, Shimadzu). A three-point bending test (12-mm span) was carried out using a universal testing machine (EZ Test, Shimadzu). Hardness and elastic modulus of external and cross-sectioned surfaces were obtained by nanoindentation (ENT-1100a, Elionix; n  =  10). Coatings on Woowa and BioForce High Aesthetic Archwire contained 41% silver and 14% gold, respectively. The coating thickness on Woowa was approximately 10 µm, and the coating thickness on BioForce High Aesthetic Archwire was much smaller. The surfaces of both coated wires were rougher than the noncoated wires. Woowa showed a higher mean unloading force than the noncoated Woowa, although BioForce High Aesthetic Archwire showed a lower mean unloading force than Sentalloy. While cross-sectional surfaces of all wires had similar hardness and elastic modulus, values for the external surface of Woowa were smaller than for the other wires. The coating processes for Woowa and BioForce High Aesthetic Archwire influence bending behavior and surface morphology.

  3. Comparison of Superelasticity of Nickel Titanium Orthodontic Arch wires using Mechanical Tensile Testing and Correlating with Electrical Resistivity

    Science.gov (United States)

    Sivaraj, Aravind

    2013-01-01

    Background: Application of light and continuous forces for optimum physiological response and least damage to the tooth supporting structures should be the primary aim of the orthodontist. Nickel titanium alloys with the properties of excellent spring back, super elasticity and wide range of action is one of the natural choices for the clinicians to achieve this goal. In recent periods, various wire manufacturers have come with a variety of wires exhibiting different properties. It is the duty of the clinician to select appropriate wires during various stages of treatment for excellent results. For achieving this evaluation of the properties of these wires is essential. Materials & Methods: This study is focussed on evaluating the super elastic property of eight groups of austenite active nickel titanium wires. Eight groups of archwires bought from eight different manufacturers were studied. These wires were tested through mechanical tensile testing and electrical resistivity methods. Results: Unloading curves were carefully assessed for superelastic behaviour on deactivation. Rankings of the wires tested were based primarily upon the unloading curve’s slope Conclusion: Ortho organisers wires ranked first and superior, followed by American Orthodontics and Ormco A wires. Morelli and GAClowland NiTi wires were ranked last. It can be concluded that the performance of these wires based on rankings should be further evaluated by clinical studies. How to cite this article: Sivaraj A. Comparison of Superelasticity of Nickel Titanium Orthodontic Arch wires using Mechanical Tensile Testing and Correlating with Electrical Resistivity. J Int Oral Health 2013; 5(3):1-12. PMID:24155596

  4. Susceptibility to corrosion and in vitro biocompatibility of a laser-welded composite orthodontic arch wire.

    Science.gov (United States)

    Zhang, Chao; Sun, Xinhua; Zhao, Shuang; Yu, Wenwen; Sun, Daqian

    2014-01-01

    Composite arch-wire (CoAW) is an arch wire formed by solder connection of nickel titanium shape memory alloy and stainless steel wire. The purpose of the present study was to investigate the biocompatibility of CoAW as an important foundation for its clinical application. The electrochemical corrosion and ion release behavior of CoAW upon immersion in solutions simulating oral cavity conditions were measured to evaluate the corrosion behavior of CoAW. Murine L-929 cells were co-cultured with CoAW extract to evaluate the cytotoxicity of the corrosion products in vitro. Polarization tests indicated that CoAW is resistant to corrosion in the tested artificial saliva (AS)-based solutions (chloric solution, simple AS, fluorinated AS, and protein-containing AS), and the amount of toxic copper ions released after immersion was lower than average daily dietary intake levels. The cytotoxicity experiments demonstrated the in vitro biocompatibility of CoAW. Based on the combined advantages of its base materials CoAW, with its resistance to biocorrosion and in vitro cytocompatibility, is a promising alternative material for use in orthodontic fixation applications.

  5. Frictional resistance of orthodontic wires tied with 3 types of elastomeric ligatures

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    Amanda Carneiro da Cunha

    2011-12-01

    Full Text Available The aims of this study were to determine and compare frictional resistance obtained by low-friction and conventional elastomeric ligatures in the presence of artificial saliva, and observe whether this variable changed after 21 days. Super Slick® low-friction elastomeric ligatures and conventional ligatures of the brands TP conventional® and Unitek® were placed on standard edgewise maxillary central incisor metal brackets, slot .022" × .028" tying rectangular orthodontic wires .018" × .025". Three experimental groups were arranged according to the type of ligature and a control group in which no wires were used. The friction values obtained between the bracket/wire/ligature set were measured using a Universal Test Machine at a speed of 20 mm/minute, at two experimental time intervals: T0 - immediately after specimen fabrication; and T1 - 21 days after fabrication and immersion in artificial saliva at 37 ºC. Conventional Unitek ligatures and the low-friction ligature (Super Slick showed the lowest friction values at T0. After 21 days (T1, however, conventional Unitek ligatures presented the lowest value. All groups assessed from T0 to T1 showed a numerical reduction in friction values, suggesting that time, heat and humidity may cause elastic degradation, however this was not verified statistically (P > 0.05.

  6. Evolution of flexural rigidity according to the cross-sectional dimension of a superelastic nickel titanium orthodontic wire.

    Science.gov (United States)

    Garrec, Pascal; Tavernier, Bruno; Jordan, Laurence

    2005-08-01

    The choice of the most suitable orthodontic wire for each stage of treatment requires estimation of the forces generated. In theory, the selection of wire sequences should initially utilize a lower flexural rigidity; thus clinicians use smaller round cross-sectional dimension wires to generate lighter forces during the preliminary alignment stage. This assessment is true for conventional alloys, but not necessarily for superelastic nickel titanium (NiTi). In this case, the flexural rigidity dependence on cross-sectional dimension differs from the linear elasticity prediction because of the martensitic transformation process. It decreases with increasing deflection and this phenomenon is accentuated in the unloading process. This behaviour should lead us to consider differently the biomechanical approach to orthodontic treatment. The present study compared bending in 10 archwires made from NiTi orthodontics alloy of two cross-sectional dimensions. The results were based on microstructural and mechanical investigations. With conventional alloys, the flexural rigidity was constant for each wire and increased largely with the cross-sectional dimension for the same strain. With NiTi alloys, the flexural rigidity is not constant and the influence of size was not as important as it should be. This result can be explained by the non-constant elastic modulus during the martensite transformation process. Thus, in some cases, treatment can begin with full-size (rectangular) wires that nearly fill the bracket slot with a force application deemed to be physiologically desirable for tooth movement and compatible with patient comfort.

  7. Evaluation of tensile strength and surface topography of orthodontic wires after infection control procedures: An in vitro study.

    Science.gov (United States)

    Brindha, M; Kumaran, N Kurunji; Rajasigamani, K

    2014-07-01

    The aim of this study is to evaluate, the influence of four types of sterilization/disinfection procedures (autoclave, hot air oven, glutaraldehyde, and ultraviolet [UV] light) on the tensile strength and surface topography of three orthodontic wires (stainless steel (SS), titanium - molybdenum alloy [TMA], and cobalt chromium (CoCr)). Sample comprised of three types of 8 inches straight length segments of orthodontic wires. They were divided into three groups according to wire composition comprising of 50 samples each. Totally 50 samples of each group were then equally divided into five subgroups according to sterilization method. After sterilization and disinfection of the experimental group, surface topography was examined with scanning electron microscope (SEM) and tensile strength was tested using universal testing machine. The results of this study show that the mean ultimate tensile strength (UTS) of SS wire after four sterilization procedures were similar to the control group (1845.815 ± 142.29 MPa). The mean UTS of TMA wire increases after four sterilization procedures when compared with the control group (874.107 ± 275.939 MPa). The mean UTS of CoCr wire remains same after UV light disinfection, but increases after other three sterilization procedures when compared with the control group (1449.759 ± 156.586 MPa). SEM photographs of the present study shows gross increase in pitting roughness of the surface topography of all the three types of wires after four types of sterilization. Orthodontists who want to offer maximum safety for their patients can sterilize orthodontic wires before placement, as it does not deteriorate the tensile strength and surface roughness of the alloys.

  8. Interceptive Correction of Anterior Crossbite Using Short-Span Wire-Fixed Orthodontic Appliance: A Report of Three Cases

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    S. Nagarajan M. P. Sockalingam

    2018-01-01

    Full Text Available Anterior crossbite is relatively a common presentation in the mixed dentition stage. If left untreated, it can lead to a host of problems and may complicate future orthodontic treatment. One of the major difficulties in performing anterior crossbite correction in young children is treatment compliance. In most cases, poor compliance is due to the unacceptability of the removable appliance used. This article describes three cases of successful correction of anterior crossbite of patients in mixed dentition using short-span wire-fixed orthodontic appliances. This sectional appliance provides an alternative method of correcting anterior crossbite of dental origin and offers many advantages compared to the use of removable appliances.

  9. A Comparison of pical Root Resorption in Incisors after Fixed Orthodontic Treatment with Standard Edgewise and Straight Wire (MBT) Method.

    Science.gov (United States)

    Zahed Zahedani, Sm; Oshagh, M; Momeni Danaei, Sh; Roeinpeikar, Smm

    2013-09-01

    One of the major outcomes of orthodontic treatment is the apical root resorption of teeth moved during the treatment. Identifying the possible risk factors, are necessary for every orthodontist. The aim of this study was to compare the rate of apical root resorption after fixed orthodontic treatment with standard edgewise and straight wire (MBT) method, and also to evaluate other factors effecting the rate of root resorption in orthodontic treatments. In this study, parallel periapical radiographs of 127 patients imaging a total of 737 individual teeth, were collected. A total of 76 patients were treated by standard edgewise and 51 patients by straight wire method. The periapical radiographs were scanned and then the percentage of root resorption was calculated by Photoshop software. The data were analyzed by Paired-Samples t-test and the Generalized Linear Model adopting the SPSS 15.0. In patients treated with straight wire method (MBT), mean root resorption was 18.26% compared to 14.82% in patients treated with standard edgewise technique (proot resorption,statistically significant (proot resorption in this study. Having more root resorption in the straight wire method and less in the standard edgewise technique can be attributed to more root movement in pre-adjusted MBT technique due to the brackets employed in this method.

  10. Do Mechanical and Physicochemical Properties of Orthodontic NiTi Wires Remain Stable In Vivo?

    Directory of Open Access Journals (Sweden)

    Michał Sarul

    2016-01-01

    Full Text Available Introduction and Aim. Exceptional properties of the NiTi archwires may be jeopardized by the oral cavity; thus its long-term effect on the mechanical and physiochemical properties of NiTi archwires was the aim of work. Material and Methods. Study group comprised sixty 0.016 × 0.022 NiTi archwires from the same manufacturer evaluated (group A after the first 12 weeks of orthodontic treatment. 30 mm long pieces cut off from each wire prior to insertion formed the control group B. Obeying the strict rules of randomization, all samples were subjected to microscopic evaluation and nanoindentation test. Results. Both groups displayed substantial presence of nonmetallic inclusions. Heterogeneity of the structure and its alteration after usage were found in groups B and A, respectively. Conclusions. Long-term, reliable prediction of biomechanics of NiTi wires in vivo is impossible, especially new archwires from the same vendor display different physiochemical properties. Moreover, manufacturers have to decrease contamination in the production process in order to minimize risk of mutual negative influence of nickel-titanium archwires and oral environment.

  11. Effects of nanostructured, diamondlike, carbon coating and nitrocarburizing on the frictional properties and biocompatibility of orthodontic stainless steel wires.

    Science.gov (United States)

    Zhang, Hao; Guo, Shuyu; Wang, Dongyue; Zhou, Tingting; Wang, Lin; Ma, Junqing

    2016-09-01

    To evaluate and compare the effects of nanostructured, diamondlike, carbon (DLC) coating and nitrocarburizing on the frictional properties and biocompatibility of orthodontic stainless steel archwires. Plasma-enhanced chemical vapor deposition technology was applied to coat DLC films onto the surface of austenitic stainless steel wires, and salt-bath nitrocarburizing technology was employed to achieve surface hardening of other wires. Surface and cross-sectional characteristics, microhardness, modulus of elasticity, friction resistance, corrosion resistance, and cell toxicity of the modified and control wires were analyzed. The surfaces of the DLC-coated and nitrocarburized wires were both smooth and even. Compared with the control, the DLC-coated wires were increased in surface hardness 1.46 times, decreased in elastic modulus, reduced in kinetic friction coefficient by 40.71%, and decreased in corrosion current density by two orders of magnitude. The nitrocarburized wire was increased in surface hardness 2.39 times, exhibited an unchanged elastic modulus, demonstrated a decrease in maximum static friction force of 22.2%, and rose in corrosion current density two orders of magnitude. Cytotoxicity tests revealed no significant toxicity associated with the modified wires. DLC coating and nitrocarburizing significantly improved the surface hardness of the wires, reduced friction, and exhibited good biocompatibility. The nanostructured DLC coating provided excellent corrosion resistance and good elasticity, and while the nitrocarburizing technique substantially improved frictional properties, it reduced the corrosion resistance of the stainless steel wires to a lesser extent.

  12. Fatigue of orthodontic nickel-titanium (NiTi) wires in different fluids under constant mechanical stress

    International Nuclear Information System (INIS)

    Prymak, O.; Klocke, A.; Kahl-Nieke, B.; Epple, M.

    2003-01-01

    The aim of this study was to analyze in vitro the fatigue resistance of nickel-titanium (NiTi) and CuNiTi orthodontic wires when subjected to forces and fluids which are present intraorally. The wires were subjected to dynamic mechanical analysis (DMA) while they were immersed into different fluids with mechanical loading parameters similar to those that are subjected in the mouth. The characteristic temperatures of transitions and a rough surface structure on the perimeter of the wires were determined by differential scanning calorimetry (DSC) and scanning electron microscopy (SEM), respectively, before and after the DMA experiments. Stainless steel wires were used for comparison. In general, NiTi wires fractured earlier than the stainless steel specimens. Survival times were lower for the NiTi wires when immersed in fluids (water, citric acid, NaCl solution, artificial saliva, and fluoridated artificial saliva) than in air. SEM surface analysis showed that the NiTi and CuNiTi wires had a rougher surface than steel wires. The fracture occurred within a short number of loading cycles. Until fracture occurred, the mechanical properties remained mostly constant

  13. Fatigue of orthodontic nickel-titanium (NiTi) wires in different fluids under constant mechanical stress

    Energy Technology Data Exchange (ETDEWEB)

    Prymak, O.; Klocke, A.; Kahl-Nieke, B.; Epple, M

    2003-07-25

    The aim of this study was to analyze in vitro the fatigue resistance of nickel-titanium (NiTi) and CuNiTi orthodontic wires when subjected to forces and fluids which are present intraorally. The wires were subjected to dynamic mechanical analysis (DMA) while they were immersed into different fluids with mechanical loading parameters similar to those that are subjected in the mouth. The characteristic temperatures of transitions and a rough surface structure on the perimeter of the wires were determined by differential scanning calorimetry (DSC) and scanning electron microscopy (SEM), respectively, before and after the DMA experiments. Stainless steel wires were used for comparison. In general, NiTi wires fractured earlier than the stainless steel specimens. Survival times were lower for the NiTi wires when immersed in fluids (water, citric acid, NaCl solution, artificial saliva, and fluoridated artificial saliva) than in air. SEM surface analysis showed that the NiTi and CuNiTi wires had a rougher surface than steel wires. The fracture occurred within a short number of loading cycles. Until fracture occurred, the mechanical properties remained mostly constant.

  14. Orthodontic buccal tooth movement by nickel-free titanium-based shape memory and superelastic alloy wire.

    Science.gov (United States)

    Suzuki, Akihiro; Kanetaka, Hiroyasu; Shimizu, Yoshinaka; Tomizuka, Ryo; Hosoda, Hideki; Miyazaki, Shuichi; Okuno, Osamu; Igarashi, Kaoru; Mitani, Hideo

    2006-11-01

    To examine the mechanical properties and the usefulness of titanium-niobium-aluminum (Ti-Nb-Al) wire in orthodontic tooth movement as compared with nickel-titanium (Ni-Ti) wire. The load deflection of expansion springs was gauged with an original jig. The gradient of the superelastic region was measured during the unloading process. Expansion springs comprising the two types of alloy wires were applied to upper first molars of rats. The distance between the first molars was measured with micrometer calipers. The force magnitude of the Ti-Nb-Al expansion spring was lower than that of the Ni-Ti expansion spring over the entire deflection range. The initial force magnitude and the gradient in the superelastic region of the Ti-Nb-Al expansion springs were half those of the Ni-Ti expansion springs. Thus, Ti-Nb-Al expansion springs generated lighter and more continuous force. Tooth movement in the Ni-Ti group proceeded in a stepwise fashion. On the other hand, tooth movement in the Ti-Nb-Al group showed relatively smooth and continuous progression. At 17 days after insertion of expansion springs, there were no significant differences between the Ti-Nb-Al and Ni-Ti groups in the amount of tooth movement. These results indicate that Ti-Nb-Al wire has excellent mechanical properties for smooth, continuous tooth movement and suggest that Ti-Nb-Al wire may be used as a practical nickel-free shape memory and superelastic alloy wire for orthodontic treatment as a substitute for Ni-Ti wire.

  15. The effect of ZnO nanoparticle coating on the frictional resistance between orthodontic wires and ceramic brackets

    Science.gov (United States)

    Behroozian, Ahmad; Kachoei, Mojgan; Khatamian, Masumeh; Divband, Baharak

    2016-01-01

    Background. Any decrease in friction between orthodontic wire and bracket can accelerate tooth movement in the sliding technique and result in better control of anchorage. This study was carried out to evaluate frictional forces by coating orthodontic wires and porcelain brackets with zinc oxide nanoparticles (ZnO). Methods. In this in vitro study, we evaluated a combination of 120 samples of 0.019×0.025 stainless steel (SS) orthodonticwires and 22 mil system edgewise porcelain brackets with and without spherical zinc oxide nanoparticles. Spherical ZnOnanoparticles were deposited on wires and brackets by immersing them in ethanol solution and SEM (scanning electronmicroscope) evaluation confirmed the presence of the ZnO coating. The frictional forces were calculated between the wiresand brackets in four groups: group ZZ (coated wire and bracket), group OO (uncoated wire and bracket), group ZO (coatedwire and uncoated bracket) and group OZ (uncoated wire and coated bracket). Kolmogorov-Smirnov, Mann-Whitney andKruskal-Wallis tests were used for data analysis. Results. The frictional force in ZZ (3.07±0.4 N) was the highest (P <0.05), and OZ (2.18±0.5 N) had the lowest amount of friction (P <0.05) among the groups. There was no significant difference in frictional forces between the ZO and OO groups (2.65±0.2 and 2.70±0.2 N, respectively). Conclusion. Coating of porcelain bracket surfaces with ZnO nanoparticles can decrease friction in the sliding technique,and wire coating combined with bracket coating is not recommended due to its effect on friction. PMID:27429727

  16. Comparison of the initial orthodontic force systems produced by a new lingual bracket system and a straight-wire appliance.

    Science.gov (United States)

    Fuck, Lars-Michael; Wiechmann, Dirk; Drescher, Dieter

    2005-09-01

    Over the last few years, lingual appliances have become an established orthodontic treatment technique. Many studies have concentrated on various esthetic aspects, on laboratory and clinical procedures, and on patient comfort and compliance. The orthodontic force systems of these appliances, however, have not yet been investigated. The aim of this study was thus to determine the forces and moments produced by a new lingual bracket system during the leveling phase of orthodontic treatment and to compare those with the corresponding force system of a labial straight-wire appliance. The intra-oral situation of ten patients undergoing orthodontic treatment was replicated in measurement casts fitted with lingual and labial brackets. Special care was taken to precisely reproduce each patient's interbracket geometry. We measured each tooth's force systems as generated by a leveling arch inserted into the lingual and labial brackets. The resulting force systems of both appliances were found to be quite similar with regard to the magnitude of most force and moment components. Only the first molars were subjected to considerably greater single forces with the lingual appliance. Tipping moments were found to be significantly smaller with the lingual technique, whereas the rotational moments were significantly smaller with the labial appliance. All in all we noted significant differences between the two techniques only in certain areas which upon closer examination were distributed over only a few tooth types. The initial force systems produced by the new lingual bracket system proved to be comparable with those delivered by a conventional straight-wire appliance. The actual levels of forces and moments, however, were found in certain cases to be too heavy with both techniques. We therefore recommend the development of leveling wires producing considerably lighter forces and moments.

  17. Evaluation of the force generated by gradual deflection of orthodontic wires in conventional metallic, esthetic, and self-ligating brackets.

    Science.gov (United States)

    Francisconi, Manoela Fávaro; Janson, Guilherme; Henriques, José Fernando Castanha; Freitas, Karina Maria Salvatore de

    2016-01-01

    The purpose of this study was to evaluate the deflection forces of Nitinol orthodontic wires placed in different types of brackets: metallic, reinforced polycarbonate with metallic slots, sapphire, passive and active self-ligating, by assessing strength values variation according to gradual increase in wire diameter and deflection and comparing different combinations in the different deflections. Specimens were set in a clinical simulation model and evaluated in a Universal Testing Machine (INSTRON 3342), using the ISO 15841 protocol. Data were subjected to One-way ANOVA, followed by Tukey tests (pbrackets presented the most similar behavior to each other. For conventional brackets there was no consistent behavior for any of the deflections studied. Self-ligating brackets presented the most consistent and predictable results while conventional brackets, as esthetic brackets, showed very different patterns of forces. Self-ligating brackets showed higher strength in all deflections when compared with the others, in 0.020-inch wires.

  18. X-ray diffraction study of low-temperature phase transformations in nickel-titanium orthodontic wires.

    Science.gov (United States)

    Iijima, M; Brantley, W A; Guo, W H; Clark, W A T; Yuasa, T; Mizoguchi, I

    2008-11-01

    Employ conventional X-ray diffraction (XRD) to analyze three clinically important nickel-titanium orthodontic wire alloys over a range of temperatures between 25 and -110 degrees C, for comparison with previous results from temperature-modulated differential scanning calorimetry (TMDSC) studies. The archwires selected were 35 degrees C Copper Ni-Ti (Ormco), Neo Sentalloy (GAC International), and Nitinol SE (3M Unitek). Neo Sentalloy, which exhibits superelastic behavior, is marketed as having shape memory in the oral environment, and Nitinol SE and 35 degrees C Copper Ni-Ti also exhibit superelastic behavior. All archwires had dimensions of 0.016in.x0.022in. (0.41 mm x 0.56 mm). Straight segments cut with a water-cooled diamond saw were placed side-by-side to yield a 1 cm x 1cm test sample of each wire product for XRD analysis (Rint-Ultima(+), Rigaku) over a 2theta range from 30 degrees to 130 degrees and at successive temperatures of 25, -110, -60, -20, 0 and 25 degrees C. The phases revealed by XRD at the different analysis temperatures were in good agreement with those found in previous TMDSC studies of transformations in these alloys, in particular verifying the presence of R-phase at 25 degrees C. Precise comparisons are not possible because of the approximate nature of the transformation temperatures determined by TMDSC and the preferred crystallographic orientation present in the wires. New XRD peaks appear to result from low-temperature transformation in martensite, which a recent transmission electron microscopy (TEM) study has shown to arise from twinning. While XRD is a useful technique to study phases in nickel-titanium orthodontic wires and their transformations as a function of temperature, optimum insight is obtained when XRD analyses are combined with complementary TMDSC and TEM study of the wires.

  19. Effect of Punica granatum L. Flower Water Extract on Five Common Oral Bacteria and Bacterial Biofilm Formation on Orthodontic Wire.

    Science.gov (United States)

    Vahid Dastjerdi, Elahe; Abdolazimi, Zahra; Ghazanfarian, Marzieh; Amdjadi, Parisa; Kamalinejad, Mohammad; Mahboubi, Arash

    2014-12-01

    Use of herbal extracts and essences as natural antibacterial compounds has become increasingly popular for the control of oral infectious diseases. Therefore, finding natural antimicrobial products with the lowest side effects seems necessary. The present study sought to assess the effect of Punica granatum L. water extract on five oral bacteria and bacterial biofilm formation on orthodontic wire. Antibacterial property of P. granatum L. water extract was primarily evaluated in brain heart infusion agar medium using well-plate method. The minimum inhibitory concentration and minimum bactericidal concentration were determined by macro-dilution method. The inhibitory effect on orthodontic wire bacterial biofilm formation was evaluated using viable cell count in biofilm medium. At the final phase, samples were fixed and analyzed by Scanning Electron Microscopy. The growth inhibition zone diameter was proportional to the extract concentration. The water extract demonstrated the maximum antibacterial effect on Streptococcus sanguinis ATCC 10556 with a minimum inhibitory concentration of 6.25 mg/ml and maximum bactericidal effect on S. sanguinis ATCC 10556 and S. sobrinus ATCC 27607 with minimum bactericidal concentration of 25 mg/ml. The water extract decreased bacterial biofilm formation by S. sanguinis, S. sobrinus, S. salivarius, S. mutans ATCC 35608 and E. faecalis CIP 55142 by 93.7-100%, 40.6-99.9%, 85.2-86.5%, 66.4-84.4% and 35.5-56.3% respectively. Punica granatum L. water extract had significant antibacterial properties against 5 oral bacteria and prevented orthodontic wire bacterial biofilm formation. However, further investigations are required to generalize these results to the clinical setting.

  20. Effect of Punica granatum L. Flower Water Extract on Five Common Oral Bacteria and Bacterial Biofilm Formation on Orthodontic Wire

    Science.gov (United States)

    VAHID DASTJERDI, Elahe; ABDOLAZIMI, Zahra; GHAZANFARIAN, Marzieh; AMDJADI, Parisa; KAMALINEJAD, Mohammad; MAHBOUBI, Arash

    2014-01-01

    Background: Use of herbal extracts and essences as natural antibacterial compounds has become increasingly popular for the control of oral infectious diseases. Therefore, finding natural antimicrobial products with the lowest side effects seems necessary. The present study sought to assess the effect of Punica granatum L. water extract on five oral bacteria and bacterial biofilm formation on orthodontic wire. Methods: Antibacterial property of P. granatum L. water extract was primarily evaluated in brain heart infusion agar medium using well-plate method. The minimum inhibitory concentration and minimum bactericidal concentration were determined by macro-dilution method. The inhibitory effect on orthodontic wire bacterial biofilm formation was evaluated using viable cell count in biofilm medium. At the final phase, samples were fixed and analyzed by Scanning Electron Microscopy. Results: The growth inhibition zone diameter was proportional to the extract concentration. The water extract demonstrated the maximum antibacterial effect on Streptococcus sanguinis ATCC 10556 with a minimum inhibitory concentration of 6.25 mg/ml and maximum bactericidal effect on S. sanguinis ATCC 10556 and S. sobrinus ATCC 27607 with minimum bactericidal concentration of 25 mg/ml. The water extract decreased bacterial biofilm formation by S. sanguinis, S. sobrinus, S. salivarius, S. mutans ATCC 35608 and E. faecalis CIP 55142 by 93.7–100%, 40.6–99.9%, 85.2–86.5%, 66.4–84.4% and 35.5–56.3% respectively. Conclusion: Punica granatum L. water extract had significant antibacterial properties against 5 oral bacteria and prevented orthodontic wire bacterial biofilm formation. However, further investigations are required to generalize these results to the clinical setting. PMID:26171362

  1. Effect of Different Types of Toothpaste on the Frictional Resistance Between Orthodontic Stainless Steel Brackets and Wires.

    Science.gov (United States)

    Hosseinzadeh Nik, Tahereh; Hooshmand, Tabassom; Farhadifard, Homa

    2017-09-01

    The purpose of this study was to investigate the effect of different types of toothpaste on the frictional resistance between stainless steel brackets and archwires. Ninety stainless steel orthodontic brackets with stainless steel wires were bonded to bovine teeth and were divided into 6 groups for application of the following toothpastes: Colgate® Total® Advanced Whitening, Colgate® Total® Pro Gum Health, Colgate® Anticavity, Ortho.Kin®, and Sunstar GUM® Ortho toothpastes. No toothpaste was applied in the control group. Each group was brushed by a brushing machine with the use of the designated solution for 4.5 minutes. The frictional force was measured in a universal testing machine with a crosshead speed of 10 mm/minute over a 5-mm archwire. Data were analyzed using one-way analysis of variance (ANOVA) at the 0.05 significance level. The frictional resistance values of Ortho.Kin® and GUM® Ortho toothpastes and the control group were not significantly different (P>0.05). However, there were significant differences between the frictional resistance values of Colgate® Total® Pro Gum Health and Colgate® Anticavity toothpastes with that of the control group (Porthodontic toothpastes did not increase the frictional resistance between the orthodontic stainless steel brackets and wires.

  2. Effect of time and pH on physical-chemical properties of orthodontic brackets and wires.

    Science.gov (United States)

    Dos Santos, Aretha Aliny Ramos; Pithon, Matheus Melo; Carlo, Fabíola Galbiatti Carvalho; Carlo, Hugo Lemes; de Lima, Bruno Alessandro Silva Guedes; Dos Passos, Tibério Andrade; Lacerda-Santos, Rogério

    2015-03-01

    To test the hypothesis that treatment time, debris/biofilm, and oral pH have an influence on the physical-chemical properties of orthodontic brackets and arch wires. One hundred twenty metal brackets were evaluated. They were divided into four groups (n  =  30) according to treatment time: group C (control) and groups T12, T24, and T36 (brackets recovered after 12, 24, and 36 months of treatment, respectively). Rectangular stainless-steel arch wires that remained in the oral cavity for 12 to 24 months were also analyzed. Dimensional stability, surface morphology, composition of brackets, resistance to sliding of the bracket-wire set, surface roughness of wires, and oral pH were analyzed. One-way analysis of variance, followed by a Tukey multiple comparisons test, was used for statistical analysis (P bracket slots were shown to have more influence on the degradation process and frictional force of these devices than did oral pH.

  3. Radiographic comparison of apical root resorption after orthodontic treatment between bidimensional and Roth straight-wire techniques.

    Science.gov (United States)

    Zawawi, Khalid H; Malki, Ghadah A

    2014-10-01

    The aim of this study was to compare the amount of root resorption after orthodontic treatment between the bidimensional and the Roth straight-wire techniques. Another objective was to compare the amount of root resorption in the whole sample studied and record the prevalence of root resorption. The sample consisted of 40 patients (age ranged between 11 and 18 years) with Angle Class II division 1 malocclusions, treated nonextraction. Twenty patients were treated with bidimensional technique and 20 with a 0.018-inch Roth straight-wire technique. Root lengths of the maxillary incisors were measured on pre- and post-treatment periapical radiographs. The results demonstrated that the bidimensional and Roth straight-wire groups showed significant root resorption after treatment, 1.11 (0.17) and 0.86 (0.05), respectively, P root shortening between the bidimensional and Roth straight-wire groups, there was no significant difference between the mean change from pre- to post-treatment between bidimensional group (mean = 1.00 ± 1.34) and Roth straight-wire group (mean = 0.88 ± 0.86), P = 0.63. Considering the whole sample, there was no root resoprtion in 32.5% of the analysed teeth. There was only mild resorption in 56.2%, moderate in 8.8% and severe in only 2.5% of the teeth. Treatment with the bidimensional technique did not produce an increase in the amount of root resorption. The prevalence and amount of root resorption was similar between bidimensional and Roth straight-wire techniques.

  4. DNA single strand break in fibroblast from Down syndrome patients

    International Nuclear Information System (INIS)

    Rozga, B.

    1992-01-01

    The radiosensitivity of tree trisomic (trisomia +21) strains of human fibroblasts to gamma radiation has been investigated in vitro and the causes of induction and repair of single strand DNA breaks in these cells have been estimated. The single strand breaks in DNA of normal and trisomic cells have been found to be ameliorated with an approximately equal efficiency. Repair has been found to be three times slower in trisomic cells compared to their normal relevant, most likely due to their elevated sensitivity to ionizing radiation and the following mortality of trisomic cells, and/or the potential occurrence of a great number of chromosome aberrations in cells irradiated in vitro. (author). 28 refs, 4 figs, 1 tab

  5. Evaluation of the force generated by gradual deflection of orthodontic wires in conventional metallic, esthetic, and self-ligating brackets

    Directory of Open Access Journals (Sweden)

    Manoela Fávaro Francisconi

    Full Text Available ABSTRACT Objective: The purpose of this study was to evaluate the deflection forces of Nitinol orthodontic wires placed in different types of brackets: metallic, reinforced polycarbonate with metallic slots, sapphire, passive and active self-ligating, by assessing strength values variation according to gradual increase in wire diameter and deflection and comparing different combinations in the different deflections. Material and Methods: Specimens were set in a clinical simulation model and evaluated in a Universal Testing Machine (INSTRON 3342, using the ISO 15841 protocol. Data were subjected to One-way ANOVA, followed by Tukey tests (p<0.05. Results: Self-ligating brackets presented the most similar behavior to each other. For conventional brackets there was no consistent behavior for any of the deflections studied. Conclusions: Self-ligating brackets presented the most consistent and predictable results while conventional brackets, as esthetic brackets, showed very different patterns of forces. Self-ligating brackets showed higher strength in all deflections when compared with the others, in 0.020-inch wires.

  6. Effect of Saliva on Load-Deflection Characteristics of Superelastic Nickel-Titanium Orthodontic Wires

    OpenAIRE

    Hosseinzadeh Nik, T.; Ghadirian, H.; Ahmadabadi, M. Nili; Shahhoseini, T.; Haj-Fathalian, M.

    2012-01-01

    Statement of Problem: Most published results about the features of NiTi wires are based on the mechanical laboratory tests in a dry environment.Purpose of Study: The purpose of this study was to investigate the effect of saliva on load-deflection characteristics of superelastic NiTi wires.Materials and Methods: In this experimental study, 15 wires of three kinds of NiTi wires (Sentalloy, Force I and Truflex) were prepared; five of them were tested in their as-received condition to provide bas...

  7. The effects of sterilization on the tensile strength of orthodontic wires.

    Science.gov (United States)

    Staggers, J A; Margeson, D

    1993-01-01

    The purpose of this study was to evaluate the effect of sterilization on the tensile strength of 0.016" beta-titanium, nickel titanium and stainless steel wires. Three common methods of sterilization--autoclaving, dry heat and ethylene oxide--were evaluated in three test trials involving zero, one and five sterilization cycles. For each of the test trials, five pieces each of 0.016" TMA, 0.016" Sentalloy and 0.016" Tru-chrome stainless steel wires were sterilized using a standard autoclave. Five other pieces of each of the same wires were sterilized in a dryclave, while an additional five pieces of each of the three wire types were sterilized using ethylene oxide. The ultimate tensile strengths of the wires were then determined using an Instron Universal Testing Machine. The data were compared for statistical differences using analysis of variance. The results showed that dry heat sterilization significantly increased the tensile strength of TMA wires after one cycle, but not after five cycles. Autoclaving and ethylene oxide sterilization did not significantly alter the tensile strength of TMA wires. Dry heat and autoclave sterilization also significantly increased the tensile strength of Sentalloy wires, but the mean strength after five sterilization cycles was not significantly different than after one cycle. Ethylene oxide sterilization of Sentalloy wires did not significantly alter the tensile strengths of that wire. There were no significant differences in the tensile strengths of the stainless steel wires following zero, one or five cycles for any of the sterilization methods.

  8. Clinical Efficiency of Two Sequences of Orthodontic Wires to Correct Crowding of the Lower Anterior Teeth

    Directory of Open Access Journals (Sweden)

    Cláudia Maria de Castro Serafim

    2015-01-01

    Full Text Available This study compared time to correction of mandibular anterior crowding using two arch wire sequences, one with conventional nickel-titanium (NiTi arch wires and the other with conventional and NiTi heat-activated arch wires. Twenty-two boys and girls (mean age: 16.68 ± 2.66 with moderate crowding (3–6 mm were assigned randomly to one of two groups and followed up for five months (six assessments when arch wires were changed. Time to crowding correction was analyzed statistically using the Kaplan-Meier method. Data were collected during the five-month follow-up, and time to correction was compared between groups using the log rank test. At the end of follow-up, mandibular crowding was corrected in 100% of the cases in the group treated with the sequence that included NiTi heat-activated arch wires, whereas about 30% of those treated with NiTi arch wires were not completely corrected. There was a significant difference in time to complete treatment between groups (log rank = 5.996; p<0.05. In the group treated with the sequence that included heat-activated wires, alignment and leveling of mandibular anterior teeth were completed earlier than in the group treated only with conventional NiTi arch wires. Clinical trial registration is found at RBR-7g5zng.

  9. Effect of saliva on load-deflection characteristics of superelastic nickel-titanium orthodontic wires.

    Science.gov (United States)

    Hosseinzadeh Nik, T; Ghadirian, H; Ahmadabadi, M Nili; Shahhoseini, T; Haj-Fathalian, M

    2012-01-01

    Most published results about the characteristics of NiTi wires are based on the mechanical laboratory tests on the as-received wires.The purpose of this study was to investigate the effect of saliva on load-deflection characteristics of superelastic NiTi wires. In this experimental study, 15 wires of three kinds of superelastic NiTi wires (Sentalloy, Force I and Truflex) were prepared. Five specimens of each wire were tested in the as-received condition (T0) to provide baseline information and the remaining wires were divided into two groups of five. Half of them were kept inside artificial saliva for one month (T1), while the others were kept in air (T2). After 30 days, three-point bending test was done in a dental arch model and data from selected points on the unloading phase of the generated graphs were used for statistical analysis. Force I and Truflex showed significantly greater force than Sentalloy. The load values of Truflex and Force I after one month exposed to artificial saliva (T1) decreased significantly, but Sentalloy was not affected significantly. The plateau gap values were not considerably different among T0, T1 and T2. Saliva decreased the load of Force I and Truflex significantly, but it did not have a statistically significant effect on Sentalloy.

  10. Single-strand DNA molecule translocation through nanoelectrode gaps

    International Nuclear Information System (INIS)

    Zhao Xiongce; Payne, Christina M; Cummings, Peter T; Lee, James W

    2007-01-01

    Molecular dynamics simulations were performed to investigate the translocation of single-strand DNA through nanoscale electrode gaps under the action of a constant driving force. The application behind this theoretical study is a proposal to use nanoelectrodes as a screening gap as part of a rapid genomic sequencing device. Preliminary results from a series of simulations using various gap widths and driving forces suggest that the narrowest electrode gap that a single-strand DNA can pass is ∼1.5 nm. The minimum force required to initiate the translocation within nanoseconds is ∼0.3 nN. Simulations using DNA segments of various lengths indicate that the minimum initiation force is insensitive to the length of DNA. However, the average threading velocity of DNA varies appreciably from short to long DNA segments. We attribute such variation to the different nature of drag force experienced by the short and long DNA segments in the environment. It is found that DNA molecules deform significantly to fit in the shape of the nanogap during the translocation

  11. Programmable autonomous synthesis of single-stranded DNA

    Science.gov (United States)

    Kishi, Jocelyn Y.; Schaus, Thomas E.; Gopalkrishnan, Nikhil; Xuan, Feng; Yin, Peng

    2018-02-01

    DNA performs diverse functional roles in biology, nanotechnology and biotechnology, but current methods for autonomously synthesizing arbitrary single-stranded DNA are limited. Here, we introduce the concept of primer exchange reaction (PER) cascades, which grow nascent single-stranded DNA with user-specified sequences following prescribed reaction pathways. PER synthesis happens in a programmable, autonomous, in situ and environmentally responsive fashion, providing a platform for engineering molecular circuits and devices with a wide range of sensing, monitoring, recording, signal-processing and actuation capabilities. We experimentally demonstrate a nanodevice that transduces the detection of a trigger RNA into the production of a DNAzyme that degrades an independent RNA substrate, a signal amplifier that conditionally synthesizes long fluorescent strands only in the presence of a particular RNA signal, molecular computing circuits that evaluate logic (AND, OR, NOT) combinations of RNA inputs, and a temporal molecular event recorder that records in the PER transcript the order in which distinct RNA inputs are sequentially detected.

  12. Fios ortodônticos: conhecer para otimizar a aplicação clínica Orthodontic wires: knowledge to optimize clinical application

    Directory of Open Access Journals (Sweden)

    Cátia Cardoso Abdo Quintão

    2009-12-01

    Full Text Available A grande variedade de fios ortodônticos presente no mercado pode gerar dúvidas quanto à melhor escolha para situações clínicas. Assim, o conhecimento das propriedades mecânicas dos mesmos facilita a escolha para aplicação do movimento ortodôntico na dependência da fase em que o tratamento se encontra. A evolução da tecnologia de manufatura dos fios e a elaboração de novas técnicas ortodônticas geraram a busca por uma melhor qualidade das ligas, a fim de torná-los biologicamente mais efetivos no que diz respeito aos dentes e tecidos de suporte. O presente artigo resume as principais características dos fios utilizados em Ortodontia, em relação ao histórico, propriedades mecânicas e aplicação clínica, de acordo com fases específicas de tratamento.The huge variety of orthodontic wires brands available in market might generate confusion as regard to the best choice for clinical application. Therefore mechanical properties knowledge about wires would help the professional to apply the best orthodontic technique depending on the treatment phase. The wires manufacturing evolution and the new orthodontic techniques proposed guided the market into the search for better quality alloys, in order to make them biologically more effective to teeth and support tissues. This paper aims to summarize some main characteristics of orthodontic wires related to their history, mechanical properties and clinical application as regard to individual phase of treatment.

  13. The release of nickel from orthodontic NiTi wires is increased by dynamic mechanical loading but not constrained by surface nitridation.

    Science.gov (United States)

    Peitsch, T; Klocke, A; Kahl-Nieke, B; Prymak, O; Epple, M

    2007-09-01

    The influence of dynamic mechanical loading and of surface nitridation on the nickel release from superelastic nickel-titanium orthodontic wires was investigated under ultrapure conditions. Commercially available superelastic NiTi arch wires (size 0.018 x 0.025'') without surface modification (Neo Sentalloy) and with nitrogen ion implantation surface treatment (Neo Sentalloy Ionguard) were analyzed. Mechanical loading of wire segments with a force similar to the physiological situation was performed with a frequency of 5 Hz in ultrapure water and saline solution, respectively. The release of nickel was monitored by atomic absorption spectroscopy for up to 36 days. The mechanically loaded wires released significantly more nickel ( approximately 45 ng cm(-2) d(-1)) than did nonloaded wires (<1 ng cm(-2) d(-1)). There was no statistically significant effect of the testing solution (water or NaCl) or of the surface nitridation. The total amount of released nickel was small in all cases, but may nevertheless account for the occasional clinical observations of adverse reactions during application of NiTi-based orthodontic appliances. The surface nitridation did not constrain the release of nickel from NiTi under continuous mechanical stress.

  14. Corrosion behavior of ion implanted nickel-titanium orthodontic wire in fluoride mouth rinse solutions.

    Science.gov (United States)

    Iijima, Masahiro; Yuasa, Toshihiro; Endo, Kazuhiko; Muguruma, Takeshi; Ohno, Hiroki; Mizoguchi, Itaru

    2010-01-01

    This study investigated the corrosion properties of ion implanted nickel-titanium wire (Neo Sentalloy Ionguard) in artificial saliva and fluoride mouth rinse solutions (Butler F Mouthrinse, Ora-Bliss). Non ion implanted nickel-titanium wire (Neo Sentalloy) was used as control. The anodic corrosion behavior was examined by potentiodynamic polarization measurement. The surfaces of the specimens were examined with SEM. The elemental depth profiles were characterized by XPS. Neo Sentalloy Ionguard in artificial saliva and Butler F Mouthrinse (500 ppm) had a lower current density than Neo Sentalloy. In addition, breakdown potential of Neo Sentalloy Ionguard in Ora-Bliss (900 ppm) was much higher than that of Neo Sentalloy although both wires had similar corrosion potential in Ora-Bliss (450 and 900 ppm). The XPS results for Neo Sentalloy Ionguard suggested that the layers consisted of TiO(2) and TiN were present on the surface and the layers may improve the corrosion properties.

  15. Molecular investigation of evaporation of biodroplets containing single-strand DNA on graphene surface.

    Science.gov (United States)

    Akbari, Fahimeh; Foroutan, Masumeh

    2018-02-14

    In this study, the water droplet behaviour of four different types of single-strand DNA with homogeneous base sequence on a graphene substrate during evaporation of the droplet was investigated using molecular dynamics (MD) simulation. The simulation results indicated that the evaporation depended on the DNA sequence. The observed changes can be divided into four parts: (i) vaporization mode, (ii) evaporation flux, (iii) mechanism of single-strand placement on the surface, and (iv) consideration of remaining single strands after evaporation. Our simulation observations indicated different evaporation modes for thymine biodroplets as compared to those for other biodroplets. The evaporation of the thymine biodroplets occurred with an increase in the contact angle, while that of the other biodroplets occur in a constant contact angle mode. Moreover, thymine biodroplets generate the lowest contact line compared to other single strands, and it is always placed far away from the centre of the droplets during evaporation. Investigating variations in the evaporation flux shows that thymine has the highest evaporation flux and guanine has the lowest. Moreover, during initial evaporation, the flux of evaporation increases at the triple point of the biodroplets containing thymine single strands, while it decreases in the other biodroplets. The following observation was obtained from the study of the placement of single strands on the substrate: guanine and thymine interacted slower than other single strands during evaporation with graphene, adenine single strand had a higher folding during evaporation, and guanine single strand showed the lowest end-to-end distance. The investigation of single-strand DNA after evaporation shows that adenine produces the most stable structure at the end of evaporation. In addition, cytosine is the most stretched single-strand DNA due to its lack of internal π-π stacking and hydrogen bonding. Therefore, cytosine single strand is more

  16. Assessment of the hardness of different orthodontic wires and brackets produced by metal injection molding and conventional methods

    Directory of Open Access Journals (Sweden)

    Shiva Alavi

    2017-01-01

    Conclusion: MIM orthodontic brackets exhibited hardness values much lower than those of SS orthodontic archwires and were more compatible with NiTi and beta-titanium archwires. A wide range of microhardness values has been reported for conventional orthodontic brackets and it should be considered that the manufacturing method might be only one of the factors affecting the mechanical properties of orthodontic brackets including hardness.

  17. Enzymatic Production of Monoclonal Stoichiometric Single-Stranded DNA Oligonucleotides

    Science.gov (United States)

    Ducani, Cosimo; Kaul, Corinna; Moche, Martin; Shih, William M.; Högberg, Björn

    2013-01-01

    Single-stranded oligonucleotides are important as research tools as probes for diagnostics and gene therapy. Today, production of oligonucleotides is done via solid-phase synthesis. However, the capabilities of current polymer chemistry are limited in comparison to what can be produced in biological systems. The errors in synthetic DNA increases with oligonucleotide length, and sequence diversity can often be a problem. Here, we present the Monoclonal Stoichiometric (MOSIC) method for enzymatic DNA oligonucleotide production. Using this method, we amplify oligonucleotides from clonal templates followed by digestion of a cutter-hairpin, resulting in pools of monoclonal oligonucleotides with precisely controlled relative stoichiometric ratios. We present data where MOSIC oligonucleotides, 14–378 nt long, were prepared either by in vitro rolling-circle amplification, or by amplification in Escherichia coli in the form of phagemid DNA. The formation of a DNA crystal and folding of DNA nanostructures confirmed the scalability, purity and stoichiometry of the produced oligonucleotides. PMID:23727986

  18. Enzymatic production of 'monoclonal stoichiometric' single-stranded DNA oligonucleotides.

    Science.gov (United States)

    Ducani, Cosimo; Kaul, Corinna; Moche, Martin; Shih, William M; Högberg, Björn

    2013-07-01

    Single-stranded oligonucleotides are important as research tools, as diagnostic probes, in gene therapy and in DNA nanotechnology. Oligonucleotides are typically produced via solid-phase synthesis, using polymer chemistries that are limited relative to what biological systems produce. The number of errors in synthetic DNA increases with oligonucleotide length, and the resulting diversity of sequences can be a problem. Here we present the 'monoclonal stoichiometric' (MOSIC) method for enzyme-mediated production of DNA oligonucleotides. We amplified oligonucleotides from clonal templates derived from single bacterial colonies and then digested cutter hairpins in the products, which released pools of oligonucleotides with precisely controlled relative stoichiometric ratios. We prepared 14-378-nucleotide MOSIC oligonucleotides either by in vitro rolling-circle amplification or by amplification of phagemid DNA in Escherichia coli. Analyses of the formation of a DNA crystal and folding of DNA nanostructures confirmed the scalability, purity and stoichiometry of the produced oligonucleotides.

  19. Finite-element analysis of NiTi wire deflection during orthodontic levelling treatment

    International Nuclear Information System (INIS)

    Razali, M F; Mahmud, A S; Abdullah, J; Mokhtar, N

    2016-01-01

    Finite-element analysis is an important product development tool in medical devices industry for design and failure analysis of devices. This tool helps device designers to quickly explore various design options, optimizing specific designs and providing a deeper insight how a device is actually performing. In this study, three-dimensional finite-element models of superelastic nickel-titanium arch wire engaged in a three brackets system were developed. The aim was to measure the effect of binding friction developed on wire-bracket interaction towards the remaining recovery force available for tooth movement. Uniaxial and three brackets bending test were modelled and validated against experimental works. The prediction made by the three brackets bending models shows good agreement with the experimental results. (paper)

  20. Finite-element analysis of NiTi wire deflection during orthodontic levelling treatment

    Science.gov (United States)

    Razali, M. F.; Mahmud, A. S.; Mokhtar, N.; Abdullah, J.

    2016-02-01

    Finite-element analysis is an important product development tool in medical devices industry for design and failure analysis of devices. This tool helps device designers to quickly explore various design options, optimizing specific designs and providing a deeper insight how a device is actually performing. In this study, three-dimensional finite-element models of superelastic nickel-titanium arch wire engaged in a three brackets system were developed. The aim was to measure the effect of binding friction developed on wire-bracket interaction towards the remaining recovery force available for tooth movement. Uniaxial and three brackets bending test were modelled and validated against experimental works. The prediction made by the three brackets bending models shows good agreement with the experimental results.

  1. Force level of small diameter nickel-titanium orthodontic wires ligated with different methods

    Directory of Open Access Journals (Sweden)

    Rodrigo Hitoshi Higa

    2017-08-01

    Full Text Available Abstract Background The aim of this study was to compare the deflection force in conventional and thermally activated nickel-titanium (NiTi wires in passive (Damon Q and active (Bioquick self-ligating brackets (SLB and in conventional brackets (CB tied by two different methods: elastomeric ligature (EL and metal ligature (ML. Methods Two wire diameters (0.014 and 0.016 in. and 10 specimens per group were used. The specimens were assembled in a clinical simulation device and tested in an Instron Universal Testing Machine, with a load cell of 10 N. For the testing procedures, the acrylic block representative of the right maxillary central incisor was palatally moved, with readings of the force at 0.5, 1, 2, and 3 mm, at a constant speed of 2 mm/min and temperature of 36.5 °C. Results The conventional NiTi released higher forces than the thermally activated NiTi archwires in large deflections. In general, the SLB showed lower forces, while the ML had higher forces, with both showing a similar force release behavior, constantly decreasing as the deflection decreased. The EL showed an irregular behavior. The active SLB showed smaller forces than passive, in large deflections. Conclusions The SLB and the ML exhibit standard force patterns during unloading, while the elastomeric ligatures exhibit a randomly distributed force release behavior.

  2. Regions of incompatibility in single-stranded DNA bacteriophages phi X174 and G4

    NARCIS (Netherlands)

    van der Avoort, H. G.; van der Ende, A.; van Arkel, G. A.; Weisbeek, P. J.

    1984-01-01

    The intracellular presence of a recombinant plasmid containing the intercistronic region between the genes H and A of bacteriophage phi X174 strongly inhibits the conversion of infecting single-stranded phi X DNA to parental replicative-form DNA. Also, transfection with single-stranded or

  3. Genetic analysis of RPA single-stranded DNA binding protein in Haloferax volcanii

    OpenAIRE

    Stroud, A. L.

    2012-01-01

    Replication protein A (RPA) is a single-stranded DNA-binding protein that is present in all three domains of life. The roles of RPA include stabilising and protecting single- stranded DNA from nuclease degradation during DNA replication and repair. To achieve this, RPA uses an oligosaccharide-binding fold (OB fold) to bind single- stranded DNA. Haloferax volcanii encodes three RPAs – RPA1, RPA2 and RPA3, of which rpa1 and rpa3 are in operons with genes encoding associated proteins (APs). ...

  4. Toxin MqsR Cleaves Single-Stranded mRNA with Various 5 Ends

    Science.gov (United States)

    2016-08-24

    either protein ORIGINAL RESEARCH Toxin MqsR cleaves single- stranded mRNA with various 5’ ends Nityananda Chowdhury1,*, Brian W. Kwan1,*, Louise C...in which a single 5′- GCU site was predicted to be single- stranded (ssRNA), double- stranded (dsRNA), in the loop of a stem - loop (slRNA), or in a...single- stranded 5′- GCU sites since cleavage was approximately 20- fold higher than cleavage seen with the 5′- GCU site in the stem - loop and

  5. Single-strand breaks in supercoiled DNA induced by vacuum-UV radiation in aqueous solution

    Energy Technology Data Exchange (ETDEWEB)

    Takakura, Kaoru; Ishikawa, Mitsuo; Hieda, Kotaro; Kobayashi, Katsumi; Ito, Atsushi; Ito, Takashi

    1986-09-01

    The induction of single-strand breaks in the DNA of plasmid pBR 322 by vacuum-UV radiation above 145 nm in aqueous solutions was studied in relation to the production of OH-radicals in water. The similarity and dissimilarity were examined on the wavelength dependence between the two effects. The maximum of single strand breaks at 150 nm could be explained by the action of OH-radicals derived from direct water photolysis: the maximum at 180 nm remains unexplained. There was no indication that the direct absorption of photon by the DNA molecule plays an important role in the production of single-strand breaks.

  6. Single-strand breaks in supercoiled DNA induced by vacuum-UV radiation in aqueous solution

    International Nuclear Information System (INIS)

    Takakura, Kaoru; Ishikawa, Mitsuo; Hieda, Kotaro; Kobayashi, Katsumi; Ito, Atsushi; Ito, Takashi

    1986-01-01

    The induction of single-strand breaks in the DNA of plasmid pBR 322 by vacuum-UV radiation above 145 nm in aqueous solutions was studied in relation to the production of OH-radicals in water. The similarity and dissimilarity were examined on the wavelength dependence between the two effects. The maximum of single strand breaks at 150 nm could be explained by the action of OH-radicals derived from direct water photolysis: the maximum at 180 nm remains unexplained. There was no indication that the direct absorption of photon by the DNA molecule plays an important role in the production of single-strand breaks. (author)

  7. Mammalian DNA single-strand break repair: an X-ra(y)ted affair.

    Science.gov (United States)

    Caldecott, K W

    2001-05-01

    The genetic stability of living cells is continuously threatened by the presence of endogenous reactive oxygen species and other genotoxic molecules. Of particular threat are the thousands of DNA single-strand breaks that arise in each cell, each day, both directly from disintegration of damaged sugars and indirectly from the excision repair of damaged bases. If un-repaired, single-strand breaks can be converted into double-strand breaks during DNA replication, potentially resulting in chromosomal rearrangement and genetic deletion. Consequently, cells have adopted multiple pathways to ensure the rapid and efficient removal of single-strand breaks. A general feature of these pathways appears to be the extensive employment of protein-protein interactions to stimulate both the individual component steps and the overall repair reaction. Our current understanding of DNA single-strand break repair is discussed, and testable models for the architectural coordination of this important process are presented. Copyright 2001 John Wiley & Sons, Inc.

  8. Comparison of the Load Deflection Characteristics of Esthetic and Metal Orthodontic Wires on Ceramic Brackets using Three Point Bending Test

    Directory of Open Access Journals (Sweden)

    Umal Hiralal Doshi

    2013-01-01

    Conclusion: Steel wires showed highest strength values, requiring the incorporation of loops and folds to reduce the load/deflection. NiTi and GFRPC wires produced more deflection at low levels of force, however the esthetic wire was shown to fracture and break.

  9. Microstructural studies of 35 degrees C copper Ni-Ti orthodontic wire and TEM confirmation of low-temperature martensite transformation.

    Science.gov (United States)

    Brantley, William A; Guo, Wenhua; Clark, William A T; Iijima, Masahiro

    2008-02-01

    Previous temperature-modulated differential scanning calorimetry (TMDSC) study of nickel-titanium orthodontic wires revealed a large exothermic low-temperature peak that was attributed to transformation within martensitic NiTi. The purpose of this study was to use transmission electron microscopy (TEM) to verify this phase transformation in a clinically popular nickel-titanium wire, identify its mechanism and confirm other phase transformations found by TMDSC, and to provide detailed information about the microstructure of this wire. The 35 degrees C Copper nickel-titanium wire (Ormco) with cross-section dimensions of 0.016 in. x 0.022 in. used in the earlier TMDSC investigation was selected. Foils were prepared for TEM analyses by mechanical grinding, polishing, dimpling, ion milling and plasma cleaning. Standard bright-field and dark-field TEM images were obtained, along with convergent-beam electron diffraction patterns. A cryo-stage with the electron microscope (Phillips CM 200) permitted the specimen to be observed at -187, -45, and 50 degrees C, as well as at room temperature. Microstructures were also observed with an optical microscope and a scanning electron microscope. Room temperature microstructures had randomly oriented, elongated grains that were twinned. Electron diffraction patterns confirmed that phase transformations took place over temperature ranges previously found by TMDSC. TEM observations revealed a high dislocation density and fine-scale oxide particles, and that twinning is the mechanism for the low-temperature transformation in martensitic NiTi. TEM confirmed the low-temperature peak and other phase transformations observed by TMDSC, and revealed that twinning in martensite is the mechanism for the low-temperature peak. The high dislocation density and fine-scale oxide particles in the microstructure are the result of the wire manufacturing process.

  10. Evaluation of frictional resistance and surface characteristics after immersion of orthodontic brackets and wire in different chemical solutions: A comparative in vitrostudy.

    Science.gov (United States)

    Nanjundan, Kavitha; Vimala, G

    2016-01-01

    To evaluate the changes of static and kinetic frictional forces between the brackets and wires following exposure to a soft drink, acidic food ingredient, and acidulated fluoride prophylactic agents. Two types of Roth prescription mandibular incisor brackets were used: 3M Unitek Victory stainless steel (SS) brackets (n = 40) and Transcend 6000 polycrystalline alumina (PCA) brackets (n = 40) as well as eighty 0.019 × 0.025" dimension ortho technology SS wires of 50 mm length each. Subsequently, brackets tied with SS wires divided into eight subgroups (n = 10) and were immersed in vinegar (pH = 3.5 ± 0.5), Pepsi ® (pH = 2.46), Colgate Phos-Flur mouth rinse (pH = 5.1), and artificial saliva (control group pH = 7) for 24 h. Changes in surface morphology under scanning electron microscope ×1000, surface roughness (Ra) with surface profilometer (single bracket and single wire from each subgroup), and frictional resistance using universal testing machine were evaluated. Highest mean (standard deviation) static frictional force of 2.65 (0.25) N was recorded in Pepsi ® followed by 2.57 (0.25) N, 2.40 (0.22) N, and 2.36 (0.17) N for Vinegar, Colgate Phos-Flur mouth rinse, and artificial saliva groups, respectively. In a similar order, lesser mean kinetic frictional forces obtained. PCA brackets revealed more surface deterioration and higher frictional force values than SS brackets. A significant positive correlation was observed between frictional forces and bracket slot roughness (r = 0.861 and 0.802, respectively, for static and kinetic frictional forces, p < 0.001 for both) and wire roughness (r = 0.243 and 0.242, respectively, for static and kinetic frictional forces, p < 0.05 for both). Findings may have long-term implications when acidic food substances are used during fixed orthodontic treatment. Further, in vivo studies are required to analyze the clinical effect of acidic mediums in the oral environment during orthodontic treatment.

  11. Evaluation of frictional resistance and surface characteristics after immersion of orthodontic brackets and wire in different chemical solutions: A comparative in vitrostudy

    Directory of Open Access Journals (Sweden)

    Kavitha Nanjundan

    2016-01-01

    Full Text Available Aim: To evaluate the changes of static and kinetic frictional forces between the brackets and wires following exposure to a soft drink, acidic food ingredient, and acidulated fluoride prophylactic agents. Materials and Methods: Two types of Roth prescription mandibular incisor brackets were used: 3M Unitek Victory stainless steel (SS brackets (n = 40 and Transcend 6000 polycrystalline alumina (PCA brackets (n = 40 as well as eighty 0.019 × 0.025" dimension ortho technology SS wires of 50 mm length each. Subsequently, brackets tied with SS wires divided into eight subgroups (n = 10 and were immersed in vinegar (pH = 3.5 ± 0.5, Pepsi ® (pH = 2.46, Colgate Phos-Flur mouth rinse (pH = 5.1, and artificial saliva (control group pH = 7 for 24 h. Changes in surface morphology under scanning electron microscope ×1000, surface roughness (Ra with surface profilometer (single bracket and single wire from each subgroup, and frictional resistance using universal testing machine were evaluated. Results: Highest mean (standard deviation static frictional force of 2.65 (0.25 N was recorded in Pepsi ® followed by 2.57 (0.25 N, 2.40 (0.22 N, and 2.36 (0.17 N for Vinegar, Colgate Phos-Flur mouth rinse, and artificial saliva groups, respectively. In a similar order, lesser mean kinetic frictional forces obtained. PCA brackets revealed more surface deterioration and higher frictional force values than SS brackets. A significant positive correlation was observed between frictional forces and bracket slot roughness (r = 0.861 and 0.802, respectively, for static and kinetic frictional forces, p < 0.001 for both and wire roughness (r = 0.243 and 0.242, respectively, for static and kinetic frictional forces, p < 0.05 for both. Conclusions: Findings may have long-term implications when acidic food substances are used during fixed orthodontic treatment. Further, in vivo studies are required to analyze the clinical effect of acidic mediums in the oral environment during

  12. Three-dimensional deformation of orthodontic brackets

    OpenAIRE

    Melenka, Garrett W; Nobes, David S; Major, Paul W; Carey, Jason P

    2013-01-01

    Braces are used by orthodontists to correct the misalignment of teeth in the mouth. Archwire rotation is a particular procedure used to correct tooth inclination. Wire rotation can result in deformation to the orthodontic brackets, and an orthodontic torque simulator has been designed to examine this wire?bracket interaction. An optical technique has been employed to measure the deformation due to size and geometric constraints of the orthodontic brackets. Images of orthodontic brackets are c...

  13. Stiffness and frictional resistance of a superelastic nickel-titanium orthodontic wire with low-stress hysteresis.

    Science.gov (United States)

    Liaw, Yu-Cheng; Su, Yu-Yu M; Lai, Yu-Lin; Lee, Shyh-Yuan

    2007-05-01

    Stress-induced martensite formation with stress hysteresis that changes the elasticity and stiffness of nickel-titanium (Ni-Ti) wire influences the sliding mechanics of archwire-guided tooth movement. This in-vitro study investigated the frictional behavior of an improved superelastic Ni-Ti wire with low-stress hysteresis. Improved superelastic Ni-Ti alloy wires (L & H Titan, Tomy International, Tokyo, Japan) with low-stress hysteresis were examined by using 3-point bending and frictional resistance tests with a universal test machine at a constant temperature of 35 degrees C, and compared with the former conventional austenitic-active superelastic Ni-Ti wires (Sentalloy, Tomy International). Wire stiffness levels were derived from differentiation of the polynomial regression of the unloading curves, and values for kinetic friction were measured at constant bending deflection distances of 0, 2, 3, and 4 mm, respectively. Compared with conventional Sentalloy wires, the L & H Titan wire had a narrower stress hysteresis including a lower loading plateau and a higher unloading plateau. In addition, L & H Titan wires were less stiff than the Sentalloy wires during most unloading stages. Values of friction measured at deflections of 0, 2, and 3 mm were significantly (P Sentalloy wires at all bending deflections (P <.05). Stress-induced martensite formation significantly reduced the stiffness and thus could be beneficial to decrease the binding friction of superelastic Ni-Ti wires during sliding with large bending deflections. Austenitic-active alloy wires with low-stress hysteresis and lower stiffness and friction offer significant potential for further investigation.

  14. Repair of X-ray-induced single-strand breaks by a cell-free system

    International Nuclear Information System (INIS)

    Seki, Shuji; Ikeda, Shogo; Tsutui, Ken; Teraoka, Hirobumi

    1990-01-01

    Repair of X-ray-induced single-strand breaks of DNA was studied in vitro using an exonuclease purified from mouse ascites sarcoma (SR-C3H/He) cells. X-ray-dose-dependent unscheduled DNA synthesis was primed by the exonuclease. Repair of X-ray-induced single-strand breaks in pUC19 plasmid DNA was demonstrated by agarose gel electrophoresis after incubating the damaged DNA with the exonuclease, DNA polymerase (Klenow fragment of DNA polymerase I or DNA polymerase β purified from SR-C3H/He cells), four deoxynucleoside triphosphates, ATP and DNA ligase (T4 DNA ligase or DNA ligase I purified from calf thymus). The present results suggested that the exonuclease is involved in the initiation of repair of X-ray-induced single-strand breaks in removing 3' ends of X-ray-damaged DNA. (author)

  15. Yield of single-strand breaks in the DNA of E. coli 10 msec after irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Fox, R A; Fielden, E M; Sapora, O [Institute of Cancer Research, Sutton (UK). Surrey Branch

    1976-04-01

    The rapid mixing of 0.3M alkali with a suspension of E.coli B/r 6 +- 3 and 144 +- 3 msec after irradiation with electrons (4.3 MeV, 0 to 50 krad) has been used to make a comparison of the yields of single strand breaks in the presence and absence of oxygen. No significant difference was observed between the numbers of single strand breaks appearing at 6 and 144 msec after irradiation. Assuming that mixing with alkali inactivates the cellular repair enzymes within several milliseconds, these results indicate that enzymic repair does not operate within this time scale. It seems probable that radiation chemical processes are responsible for the initial oxygen effect on single strand breaks.

  16. Electron attachment to DNA single strands: gas phase and aqueous solution.

    Science.gov (United States)

    Gu, Jiande; Xie, Yaoming; Schaefer, Henry F

    2007-01-01

    The 2'-deoxyguanosine-3',5'-diphosphate, 2'-deoxyadenosine-3',5'-diphosphate, 2'-deoxycytidine-3',5'-diphosphate and 2'-deoxythymidine-3',5'-diphosphate systems are the smallest units of a DNA single strand. Exploring these comprehensive subunits with reliable density functional methods enables one to approach reasonable predictions of the properties of DNA single strands. With these models, DNA single strands are found to have a strong tendency to capture low-energy electrons. The vertical attachment energies (VEAs) predicted for 3',5'-dTDP (0.17 eV) and 3',5'-dGDP (0.14 eV) indicate that both the thymine-rich and the guanine-rich DNA single strands have the ability to capture electrons. The adiabatic electron affinities (AEAs) of the nucleotides considered here range from 0.22 to 0.52 eV and follow the order 3',5'-dTDP > 3',5'-dCDP > 3',5'-dGDP > 3',5'-dADP. A substantial increase in the AEA is observed compared to that of the corresponding nucleic acid bases and the corresponding nucleosides. Furthermore, aqueous solution simulations dramatically increase the electron attracting properties of the DNA single strands. The present investigation illustrates that in the gas phase, the excess electron is situated both on the nucleobase and on the phosphate moiety for DNA single strands. However, the distribution of the extra negative charge is uneven. The attached electron favors the base moiety for the pyrimidine, while it prefers the 3'-phosphate subunit for the purine DNA single strands. In contrast, the attached electron is tightly bound to the base fragment for the cytidine, thymidine and adenosine nucleotides, while it almost exclusively resides in the vicinity of the 3'-phosphate group for the guanosine nucleotides due to the solvent effects. The comparatively low vertical detachment energies (VDEs) predicted for 3',5'-dADP(-) (0.26 eV) and 3',5'-dGDP(-) (0.32 eV) indicate that electron detachment might compete with reactions having high activation barriers

  17. Phenylketonuria in The Netherlands : 93% of the mutations are detected by single-strand conformation analysis

    NARCIS (Netherlands)

    vanderSijsBos, CJM; Diepstraten, CM; Juyn, JA; Plaisier, M; Giltay, JC; vanSpronsen, FJ; Smit, GPA; Berger, R; Smeitink, JAM; PollThe, BT; vanAmstel, JKP

    1996-01-01

    Single-strand conformational analysis was used to screen for genetic defects in all thirteen exons of the phenylalanine hydroxylase gene (PAH) in phenylketonuria and hyperphenylalaninemia patients in the Netherlands. Exons that showed a bandshift were sequenced directly, In this way, we were able to

  18. CHARACTERIZATION OF SINGLE-STRAND ORIGINS OF CRYPTIC ROLLING-CIRCLE PLASMIDS FROM BACILLUS-SUBTILIS

    NARCIS (Netherlands)

    MEIJER, WJJ; VENEMA, G; BRON, S

    1995-01-01

    In this paper we describe the isolation and characterization of single strand origins (SSOs) of several cryptic Bacillus subtilis plasmids which use the rolling-circle mechanism of replication, The plasmids used in this study involved pTA1015, pTA1020, pTA1030, pTA1040, pTA1050 and pTA1060, The SSO

  19. STRUCTURAL AND FUNCTIONAL-ANALYSIS OF THE SINGLE-STRAND ORIGIN OF REPLICATION FROM THE LACTOCOCCAL PLASMID PWVO1

    NARCIS (Netherlands)

    SEEGERS, JFML; ZHAO, AC; MEIJER, WJJ; KHAN, SA; VENEMA, G; BRON, S

    1995-01-01

    The single-strand origin (SSO) of the rolling-circle (RC), broad-host-range lactococcal plasmid pWVO1 was functionally characterized. The activity of this SSO in the conversion of single-stranded DNA to double-stranded DNA was tested both in vivo and in vitro. In addition, the effect of this SSO on

  20. Genetic and biochemical identification of a novel single-stranded DNA binding complex in Haloferax volcanii

    Directory of Open Access Journals (Sweden)

    Amy eStroud

    2012-06-01

    Full Text Available Single-stranded DNA binding proteins play an essential role in DNA replication and repair. They use oligosaccharide-binding folds, a five-stranded ß-sheet coiled into a closed barrel, to bind to single-stranded DNA thereby protecting and stabilizing the DNA. In eukaryotes the single-stranded DNA binding protein is known as replication protein A (RPA and consists of three distinct subunits that function as a heterotrimer. The bacterial homolog is termed single-stranded DNA-binding protein (SSB and functions as a homotetramer. In the archaeon Haloferax volcanii there are three genes encoding homologs of RPA. Two of the rpa genes (rpa1 and rpa3 exist in operons with a novel gene specific to Euryarchaeota, this gene encodes a protein that we have termed rpa-associated protein (RPAP. The rpap genes encode proteins belonging to COG3390 group and feature oligosaccharide-binding folds, suggesting that they might cooperate with RPA in binding to single-stranded DNA. Our genetic analysis showed that rpa1 and rpa3 deletion mutants have differing phenotypes; only ∆rpa3 strains are hypersensitive to DNA damaging agents. Deletion of the rpa3-associated gene rpap3 led to similar levels of DNA damage sensitivity, as did deletion of the rpa3 operon, suggesting that RPA3 and RPAP3 function in the same pathway. Protein pull-downs involving recombinant hexahistidine-tagged RPAs showed that RPA3 co-purifies with RPAP3, and RPA1 co-purifies with RPAP1. This indicates that the RPAs interact only with their respective associated proteins; this was corroborated by the inability to construct rpa1 rpap3 and rpa3 rpap1 double mutants. This is the first report investigating the individual function of the archaeal COG3390 RPA-associated proteins. We have shown genetically and biochemically that the RPAPs interact with their respective RPAs, and have uncovered a novel single-stranded DNA binding complex that is unique to Euryarchaeota.

  1. Sites of termination of in vitro DNA synthesis on psoralen phototreated single-stranded templates

    International Nuclear Information System (INIS)

    Piette, J.; Hearst, J.

    1985-01-01

    Single-stranded DNA has been photochemically induced to react with 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) and used as substrate for DNA replication with E. coli DNA polymerase I large fragment. By using the dideoxy sequencing procedure, it is possible to map the termination sites on the template photoreacted with HMT. These sites occur at the nucleotides preceding each thymine residue (and a few cytosine residues), emphasizing the fact that in a single-stranded stretch of DNA, HMT reacts with each thymine residue without any specificity regarding the flanking base sequence of the thymine residues. In addition, termination of DNA synthesis due to psoralen-adducted thymine is not influenced by the efficiency of the 3'-5' exonuclease proof-reading activity of the DNA polymerase. (author)

  2. Expression, purification and biochemical characterization of a single-stranded DNA binding protein from Herbaspirillum seropedicae.

    Science.gov (United States)

    Vernal, Javier; Serpa, Viviane I; Tavares, Carolina; Souza, Emanuel M; Pedrosa, Fábio O; Terenzi, Hernán

    2007-05-01

    An open reading frame encoding a protein similar in size and sequence to the Escherichia coli single-stranded DNA binding protein (SSB protein) was identified in the Herbaspirillum seropedicae genome. This open reading frame was cloned into the expression plasmid pET14b. The SSB protein from H. seropedicae, named Hs_SSB, was overexpressed in E. coli strain BL21(DE3) and purified to homogeneity. Mass spectrometry data confirmed the identity of this protein. The apparent molecular mass of the native Hs_SSB was estimated by gel filtration, suggesting that the native protein is a tetramer made up of four similar subunits. The purified protein binds to single-stranded DNA (ssDNA) in a similar manner to other SSB proteins. The production of this recombinant protein in good yield opens up the possibility of obtaining its 3D-structure and will help further investigations into DNA metabolism.

  3. Repair of single-strand breaks in normal and trisomic lymphocytes

    International Nuclear Information System (INIS)

    Leonard, J.C.; Merz, T.

    1982-01-01

    Recently, Athanasiou and colleagues (1981) reported a deficiency in the capacity of lymphocytes from persons with Down's syndrome to repair single-strand DNA breaks. They found that 1 h after exposure to 160 Gray, repair processes had restored the sedimentation profile of DNA from normal lymphocytes to control values, whereas the relative average molecular weight of DNA from irradiated lymphocytes from persons with Down's syndrome showed no increase during the repair interval. They have suggested that their data, in conjunction with the earlier data concerning the frequencies of induced chromosomal aberrations in lymphocytes from persons with Down's syndrome, reflect a decreased efficiency in some aspect of DNA repair in trisomic cells. However, for further studies of this hypothesis, it is more appropriate to study the rejoining of DNA single-strand breaks after doses comparable to those used in tests for chromosomal aberrations. (orig.)

  4. The Globular State of the Single-Stranded RNA: Effect of the Secondary Structure Rearrangements

    Science.gov (United States)

    Grigoryan, Zareh A.; Karapetian, Armen T.

    2015-01-01

    The mutual influence of the slow rearrangements of secondary structure and fast collapse of the long single-stranded RNA (ssRNA) in approximation of coarse-grained model is studied with analytic calculations. It is assumed that the characteristic time of the secondary structure rearrangement is much longer than that for the formation of the tertiary structure. A nonequilibrium phase transition of the 2nd order has been observed. PMID:26345143

  5. The Globular State of the Single-Stranded RNA: Effect of the Secondary Structure Rearrangements

    Directory of Open Access Journals (Sweden)

    Zareh A. Grigoryan

    2015-01-01

    Full Text Available The mutual influence of the slow rearrangements of secondary structure and fast collapse of the long single-stranded RNA (ssRNA in approximation of coarse-grained model is studied with analytic calculations. It is assumed that the characteristic time of the secondary structure rearrangement is much longer than that for the formation of the tertiary structure. A nonequilibrium phase transition of the 2nd order has been observed.

  6. Temporary electron localization and scattering in disordered single strands of DNA

    International Nuclear Information System (INIS)

    Caron, Laurent; Sanche, Leon

    2006-01-01

    We present a theoretical study of the effect of structural and base sequence disorders on the transport properties of nonthermal electron scattering within and from single strands of DNA. The calculations are based on our recently developed formalism to treat multiple elastic scattering from simplified pseudomolecular DNA subunits. Structural disorder is shown to increase both the elastic scattering cross section and the attachment probability on the bases at low energy. Sequence disorder, however, has no significant effect

  7. Defective processing of methylated single-stranded DNA by E. coli alkB mutants

    Science.gov (United States)

    Dinglay, Suneet; Trewick, Sarah C.; Lindahl, Tomas; Sedgwick, Barbara

    2000-01-01

    Escherichia coli alkB mutants are very sensitive to DNA methylating agents. Despite these mutants being the subject of many studies, no DNA repair or other function has been assigned to the AlkB protein or to its human homolog. Here, we report that reactivation of methylmethanesulfonate (MMS)-treated single-stranded DNA phages, M13, f1, and G4, was decreased dramatically in alkB mutants. No such decrease occurred when using methylated λ phage or M13 duplex DNA. These data show that alkB mutants have a marked defect in processing methylation damage in single-stranded DNA. Recombinant AlkB protein bound more efficiently to single- than double-stranded DNA. The single-strand damage processed by AlkB was primarily cytotoxic and not mutagenic and was induced by SN2 methylating agents, MMS, DMS, and MeI but not by SN1 agent N-methyl-N-nitrosourea or by γ irradiation. Strains lacking other DNA repair activities, alkA tag, xth nfo, uvrA, mutS, and umuC, were not defective in reactivation of methylated M13 phage and did not enhance the defect of an alkB mutant. A recA mutation caused a small but additive defect. Thus, AlkB functions in a novel pathway independent of these activities. We propose that AlkB acts on alkylated single-stranded DNA in replication forks or at transcribed regions. Consistent with this theory, stationary phase alkB cells were less MMS sensitive than rapidly growing cells. PMID:10950872

  8. Solubilization of Single-walled Carbon Nanotubes with Single- stranded DNA Generated from Asymmetric PCR

    Directory of Open Access Journals (Sweden)

    Chunhai Fan

    2007-07-01

    Full Text Available Carbon nanotubes (CNTs can be effectively dispersed and functionalized bywrapping with long single-stranded DNA (ssDNA synthesized by asymmetric PCR. ThessDNA-CNTs attached on surface of glass carbon electrode made it possible forelectrochemical analysis and sensing, which was demonstrated by reduction of H2O2 onhemoglobin/ssDNA-CNTs modified electrodes. This research showed the potentialapplication of DNA-functionalised CNTs in construction of future electrochemicalbiosensors.

  9. MEIOB targets single-strand DNA and is necessary for meiotic recombination.

    Directory of Open Access Journals (Sweden)

    Benoit Souquet

    Full Text Available Meiotic recombination is a mandatory process for sexual reproduction. We identified a protein specifically implicated in meiotic homologous recombination that we named: meiosis specific with OB domain (MEIOB. This protein is conserved among metazoan species and contains single-strand DNA binding sites similar to those of RPA1. Our studies in vitro revealed that both recombinant and endogenous MEIOB can be retained on single-strand DNA. Those in vivo demonstrated the specific expression of Meiob in early meiotic germ cells and the co-localization of MEIOB protein with RPA on chromosome axes. MEIOB localization in Dmc1 (-/- spermatocytes indicated that it accumulates on resected DNA. Homologous Meiob deletion in mice caused infertility in both sexes, due to a meiotic arrest at a zygotene/pachytene-like stage. DNA double strand break repair and homologous chromosome synapsis were impaired in Meiob (-/- meiocytes. Interestingly MEIOB appeared to be dispensable for the initial loading of recombinases but was required to maintain a proper number of RAD51 and DMC1 foci beyond the zygotene stage. In light of these findings, we propose that RPA and this new single-strand DNA binding protein MEIOB, are essential to ensure the proper stabilization of recombinases which is required for successful homology search and meiotic recombination.

  10. Transformation of Saccharomyces cerevisiae with UV-irradiated single-stranded plasmid.

    Science.gov (United States)

    Zgaga, Z

    1991-08-01

    UV-irradiated single-stranded replicative plasmids were used to transform different yeast strains. The low doses of UV used in this study (10-75 J/m2) caused a significant decrease in the transforming efficiency of plasmid DNA in the Rad+ strain, while they had no effect on transformation with double-stranded plasmids of comparable size. Neither the rev3 mutation, nor the rad18 or rad52 mutations influenced the efficiency of transformation with irradiated single-stranded plasmid. However, it was found to be decreased in the double rev3 rad52 mutant. Extracellular irradiation of plasmid that contains both URA3 and LEU2 genes (psLU) gave rise to up to 5% Leu- transformants among selected Ura+ ones in the repair-proficient strain. Induction of Leu- transformants was dose-dependent and only partially depressed in the rev3 mutant. These results suggest that both mutagenic and recombinational repair processes operate on UV-damaged single-stranded DNA in yeast.

  11. A neutral glyoxal gel electrophoresis method for the detection and semi-quantitation of DNA single-strand breaks.

    Science.gov (United States)

    Pachkowski, Brian; Nakamura, Jun

    2013-01-01

    Single-strand breaks are among the most prevalent lesions found in DNA. Traditional electrophoretic methods (e.g., the Comet assay) used for investigating these lesions rely on alkaline conditions to denature DNA prior to electrophoresis. However, the presence of alkali-labile sites in DNA can result in the introduction of additional single-strand breaks upon alkali treatment during DNA sample processing. Herein, we describe a neutral glyoxal gel electrophoresis assay which is based on alkali-free DNA denaturation and is suitable for qualitative and semi-quantitative analyses of single-strand breaks in DNA isolated from different organisms.

  12. Um novo fio de aço inoxidável para aplicações ortodônticas A new stainless steel wire for orthodontic purposes

    Directory of Open Access Journals (Sweden)

    André Itman Filho

    2011-08-01

    Full Text Available OBJETIVO: desenvolver uma metodologia para fabricação de fios ortodônticos de aço inoxidável austeno-ferrítico SEW 410 Nr. 14517 por meio dos processos convencionais de laminação e trefilação. MÉTODOS: o aço austeno-ferrítico foi elaborado em um forno elétrico de indução. A qualidade dos fios foi avaliada por ensaios de tração e medidas de microdureza. A ductilidade e a manuseabilidade foram analisadas por meio da confecção de componentes ortodônticos. RESULTADOS E CONCLUSÕES: os valores encontrados mostraram que os fios de aço inoxidável austeno-ferrítico atenderam às normas BS 3507:1976 e ISO 5832-1, e apresentaram ótima ductilidade para confecção de componentes ortodônticos com dobras complexas.OBJECTIVE: To develop a method to manufacture austenitic-ferritic stainless steel orthodontic wires (SEW 410 Nr. 14517 using conventional rolling and wiredrawing processes. METHODS: Austenitic-ferritic steel was produced in an induction furnace. Traction trials and microhardness measurements were used to evaluate wire quality. Orthodontic parts were fabricated to assess ductility and malleability. RESULTS AND CONCLUSIONS: Austenitic-ferritic stainless steel wires meet the BS 3507:1976 and ISO 5832-1 norms and have excellent ductility for the fabrication of orthodontic parts with complex folds.

  13. Induction and repair of double- and single-strand DNA breaks in bacteriophage lambda superinfecting Escherichia coli

    International Nuclear Information System (INIS)

    Boye, E.; Krisch, R.E.

    1980-01-01

    Induction and repair of double-and single-strand DNA breaks have been measured after decays of 125 I and 3 H incorporated into the DNA and after external irradiation with 4 MeV electrons. For the decay experiments, cells of wild type Escherichia coli K-12 were superinfected with bacteriophage lambda DNA labelled with 5'-( 125 I)iodo-2'-deoxyuridine or with (methyl- 3 H)thymidine and frozen in liquid nitrogen. Aliquots were thawed at intervals and lysed at neutral pH, and the phage DNA was assayed for double- and single-strand breakage by neutral sucrose gradient centrifugation. The gradients used allowed measurements of both kinds of breaks in the same gradient. Decays of 125 I induced 0.39 single-strand breaks per double-strand break. No repair of either break type could be detected. Each 3 H disintegration caused 0.20 single-strand breaks and very few double-strand breaks. The single-strand breaks were rapidly rejoined after the cells were thawed. For irradiation with 4 MeV electrons, cells of wild type E. coli K-12 were superinfected with phage lambda and suspended in growth medium. Irradiation induced 42 single-strand breaks per double-strand break. The rates of break induction were 6.75 x 10 -14 (double-strand breaks) and 2.82 x 10 -12 (single-strand breaks) per rad and per dalton. The single-strand breaks were rapidly repaired upon incubation whereas the double-strand breaks seemed to remain unrepaired. It is concluded that double-strand breaks in superinfecting bacteriophage lambda DNA are repaired to a very small extent, if at all. (Author)

  14. In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element Specific for Bromacil

    Directory of Open Access Journals (Sweden)

    Ryan M. Williams

    2014-01-01

    Full Text Available Bromacil is a widely used herbicide that is known to contaminate environmental systems. Due to the hazards it presents and inefficient detection methods, it is necessary to create a rapid and efficient sensing device. Towards this end, we have utilized a stringent in vitro selection method to identify single-stranded DNA molecular recognition elements (MRE specific for bromacil. We have identified one MRE with high affinity (Kd=9.6 nM and specificity for bromacil compared to negative targets of selection and other pesticides. The selected ssDNA MRE will be useful as the sensing element in a field-deployable bromacil detection device.

  15. A single-stranded architecture for cotranscriptional folding of RNA nanostructures

    DEFF Research Database (Denmark)

    Geary, Cody; Rothemund, Paul; Andersen, Ebbe Sloth

    2014-01-01

    Artificial DNA and RNA structures have been used as scaffolds for a variety of nanoscale devices. In comparison to DNA structures, RNA structures have been limited in size, but they also have advantages: RNA can fold during transcription and thus can be genetically encoded and expressed in cells....... We introduce an architecture for designing artificial RNA structures that fold from a single strand, in which arrays of antiparallel RNA helices are precisely organized by RNA tertiary motifs and a new type of crossover pattern. We constructed RNA tiles that assemble into hexagonal lattices...

  16. On the Formation of Thymine Photodimers in Thymine Single Strands and Calf Thymus DNA

    DEFF Research Database (Denmark)

    Baggesen, Lisbeth Munksgård; Hoffmann, S.V.; Nielsen, Steen Brøndsted

    2014-01-01

    a principal component analysis of the CD spectra, we extract fingerprint spectra of both the cyclobutane pyrimidine dimer (CPD) and the pyrimidine (6-4) pyrimidone photoadduct (64PP). Extending the CD measurements to the vacuum ultraviolet region in combination with systematic examinations of size effects...... of terminal thymines, i.e., the reaction does not occur preferentially at the extremities of the single strands as previously stated. It is even possible to form two dimers with only two bridging thymines. Finally, experiments conducted on calf thymus DNA provided a similar signature of the photodimer...

  17. RADX interacts with single-stranded DNA to promote replication fork stability

    DEFF Research Database (Denmark)

    Schubert, Lisa; Ho, Teresa; Hoffmann, Saskia

    2017-01-01

    Single-stranded DNA (ssDNA) regions form as an intermediate in many DNA-associated transactions. Multiple cellular proteins interact with ssDNA via the oligonucleotide/oligosaccharide-binding (OB) fold domain. The heterotrimeric, multi-OB fold domain-containing Replication Protein A (RPA) complex...... ssDNA-binding activities is critical for avoiding these defects. Our findings establish RADX as an important component of cellular pathways that promote DNA replication integrity under basal and stressful conditions by means of multiple ssDNA-binding proteins....

  18. Purification of Single-Stranded cDNA Based on RNA Degradation Treatment and Adsorption Chromatography.

    Science.gov (United States)

    Trujillo-Esquivel, Elías; Franco, Bernardo; Flores-Martínez, Alberto; Ponce-Noyola, Patricia; Mora-Montes, Héctor M

    2016-08-02

    Analysis of gene expression is a common research tool to study networks controlling gene expression, the role of genes with unknown function, and environmentally induced responses of organisms. Most of the analytical tools used to analyze gene expression rely on accurate cDNA synthesis and quantification to obtain reproducible and quantifiable results. Thus far, most commercial kits for isolation and purification of cDNA target double-stranded molecules, which do not accurately represent the abundance of transcripts. In the present report, we provide a simple and fast method to purify single-stranded cDNA, exhibiting high purity and yield. This method is based on the treatment with RNase H and RNase A after cDNA synthesis, followed by separation in silica spin-columns and ethanol precipitation. In addition, our method avoids the use of DNase I to eliminate genomic DNA from RNA preparations, which improves cDNA yield. As a case report, our method proved to be useful in the purification of single-stranded cDNA from the pathogenic fungus Sporothrix schenckii.

  19. Genomic mapping of single-stranded DNA in hydroxyurea-challenged yeasts identifies origins of replication.

    Science.gov (United States)

    Feng, Wenyi; Collingwood, David; Boeck, Max E; Fox, Lindsay A; Alvino, Gina M; Fangman, Walton L; Raghuraman, Mosur K; Brewer, Bonita J

    2006-02-01

    During DNA replication one or both strands transiently become single stranded: first at the sites where initiation of DNA synthesis occurs (known as origins of replication) and subsequently on the lagging strands of replication forks as discontinuous Okazaki fragments are generated. We report a genome-wide analysis of single-stranded DNA (ssDNA) formation in the presence of hydroxyurea during DNA replication in wild-type and checkpoint-deficient rad53 Saccharomyces cerevisiae cells. In wild-type cells, ssDNA was first observed at a subset of replication origins and later 'migrated' bi-directionally, suggesting that ssDNA formation is associated with continuously moving replication forks. In rad53 cells, ssDNA was observed at virtually every known origin, but remained there over time, suggesting that replication forks stall. Telomeric regions seemed to be particularly sensitive to the loss of Rad53 checkpoint function. Replication origins in Schizosaccharomyces pombe were also mapped using our method.

  20. Methods for the preparation of large quantities of complex single-stranded oligonucleotide libraries.

    Science.gov (United States)

    Murgha, Yusuf E; Rouillard, Jean-Marie; Gulari, Erdogan

    2014-01-01

    Custom-defined oligonucleotide collections have a broad range of applications in fields of synthetic biology, targeted sequencing, and cytogenetics. Also, they are used to encode information for technologies like RNA interference, protein engineering and DNA-encoded libraries. High-throughput parallel DNA synthesis technologies developed for the manufacture of DNA microarrays can produce libraries of large numbers of different oligonucleotides, but in very limited amounts. Here, we compare three approaches to prepare large quantities of single-stranded oligonucleotide libraries derived from microarray synthesized collections. The first approach, alkaline melting of double-stranded PCR amplified libraries with a biotinylated strand captured on streptavidin coated magnetic beads results in little or no non-biotinylated ssDNA. The second method wherein the phosphorylated strand of PCR amplified libraries is nucleolyticaly hydrolyzed is recommended when small amounts of libraries are needed. The third method combining in vitro transcription of PCR amplified libraries to reverse transcription of the RNA product into single-stranded cDNA is our recommended method to produce large amounts of oligonucleotide libraries. Finally, we propose a method to remove any primer binding sequences introduced during library amplification.

  1. The impact of base stacking on the conformations and electrostatics of single-stranded DNA.

    Science.gov (United States)

    Plumridge, Alex; Meisburger, Steve P; Andresen, Kurt; Pollack, Lois

    2017-04-20

    Single-stranded DNA (ssDNA) is notable for its interactions with ssDNA binding proteins (SSBs) during fundamentally important biological processes including DNA repair and replication. Previous work has begun to characterize the conformational and electrostatic properties of ssDNA in association with SSBs. However, the conformational distributions of free ssDNA have been difficult to determine. To capture the vast array of ssDNA conformations in solution, we pair small angle X-ray scattering with novel ensemble fitting methods, obtaining key parameters such as the size, shape and stacking character of strands with different sequences. Complementary ion counting measurements using inductively coupled plasma atomic emission spectroscopy are employed to determine the composition of the ion atmosphere at physiological ionic strength. Applying this combined approach to poly dA and poly dT, we find that the global properties of these sequences are very similar, despite having vastly different propensities for single-stranded helical stacking. These results suggest that a relatively simple mechanism for the binding of ssDNA to non-specific SSBs may be at play, which explains the disparity in binding affinities observed for these systems. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  2. Transcription blockage by homopurine DNA sequences: role of sequence composition and single-strand breaks

    Science.gov (United States)

    Belotserkovskii, Boris P.; Neil, Alexander J.; Saleh, Syed Shayon; Shin, Jane Hae Soo; Mirkin, Sergei M.; Hanawalt, Philip C.

    2013-01-01

    The ability of DNA to adopt non-canonical structures can affect transcription and has broad implications for genome functioning. We have recently reported that guanine-rich (G-rich) homopurine-homopyrimidine sequences cause significant blockage of transcription in vitro in a strictly orientation-dependent manner: when the G-rich strand serves as the non-template strand [Belotserkovskii et al. (2010) Mechanisms and implications of transcription blockage by guanine-rich DNA sequences., Proc. Natl Acad. Sci. USA, 107, 12816–12821]. We have now systematically studied the effect of the sequence composition and single-stranded breaks on this blockage. Although substitution of guanine by any other base reduced the blockage, cytosine and thymine reduced the blockage more significantly than adenine substitutions, affirming the importance of both G-richness and the homopurine-homopyrimidine character of the sequence for this effect. A single-strand break in the non-template strand adjacent to the G-rich stretch dramatically increased the blockage. Breaks in the non-template strand result in much weaker blockage signals extending downstream from the break even in the absence of the G-rich stretch. Our combined data support the notion that transcription blockage at homopurine-homopyrimidine sequences is caused by R-loop formation. PMID:23275544

  3. Helical filaments of human Dmc1 protein on single-stranded DNA: a cautionary tale

    Science.gov (United States)

    Yu, Xiong; Egelman, Edward H.

    2010-01-01

    Proteins in the RecA/Rad51/RadA family form nucleoprotein filaments on DNA that catalyze a strand exchange reaction as part of homologous genetic recombination. Because of the centrality of this system to many aspects of DNA repair, the generation of genetic diversity, and cancer when this system fails or is not properly regulated, these filaments have been the object of many biochemical and biophysical studies. A recent paper has argued that the human Dmc1 protein, a meiotic homolog of bacterial RecA and human Rad51, forms filaments on single stranded DNA with ∼ 9 subunits per turn in contrast to the filaments formed on double stranded DNA with ∼ 6.4 subunits per turn, and that the stoichiometry of DNA binding is different between these two filaments. We show using scanning transmission electron microscopy (STEM) that the Dmc1 filament formed on single stranded DNA has a mass per unit length expected from ∼ 6.5 subunits per turn. More generally, we show how ambiguities in helical symmetry determination can generate incorrect solutions, and why one sometimes must use other techniques, such as biochemistry, metal shadowing, or STEM to resolve these ambiguities. While three-dimensional reconstruction of helical filaments from EM images is a powerful tool, the intrinsic ambiguities that may be present with limited resolution are not sufficiently appreciated. PMID:20600108

  4. Influence of DNA conformation on radiation-induced single-strand breaks

    International Nuclear Information System (INIS)

    Barone, F.; Belli, M.; Mazzei, F.

    1994-01-01

    We performed experiments on two DNA fragments of about 300 bp having different conformation to test whether radiation-induced single-strand breakage is dependent on DNA conformation. Breakage analysis was carried out by denaturing polyacrylamide gel electrophoresis, which allows determination of the broken site at single nucleotide resolution. We found uniform cutting patterns in B-form regions. On the contrary, X- or γ-irradiation of curved fragments of kinetoplast DNA showed that the distribution of single-strand breaks was not uniform along the fragment, as the cleavage pattern was modulated in phase with the runs of A-T pairs. This modulation likely reflected the reduced accessibility of the sites which on hydroxyl-radical attack give rise to strand breaks. The cleavage pattern was phased with the runs of A-T pairs. Moreover, the overall yield of strand breaks was considerably lower in curved DNA fragments than in those with extended straight regions. The conformation effect found here indicates that the cleavage pattern reflects the fine structural features of DNA. (orig./MG)

  5. A novel pre-surgical maxillary orthodontic device using β-titanium wire for wide unilateral cleft lip and palate patients: preliminary study of its efficacy and impact for the maxillary formation.

    Science.gov (United States)

    Taniguchi, Maki; Oyama, Tomoki; Kiya, Koichiro; Sone, Yumiko; Ishii, Nobuyuki; Hosokawa, Ko

    2014-02-01

    For patients with a wide, complete, unilateral cleft lip and palate, pre-surgical maxillary orthodontic treatments have been used to reduce the alveolar gap before cheiloplasty. However, most of these treatments are complicated and laborious for patients and for medical professionals. Thus, we developed an original pre-surgical orthodontic device made with two separate acrylic resin plates connected with a spring-shaped β-titanium wire (β-TW). When the device was applied on the palate, each segment of the maxilla was automatically aligned for our target formation with the elastic force of β-titanium alloy. This study aimed to evaluate the efficacy of the new device and the size of the maxilla in comparison with the conventional Hotz procedures. A total of 47 patients with a wide unilateral cleft lip and palate were retrospectively evaluated; 33 patients were treated with our new device (β-TW plate group) and 14 were treated with a Hotz plate (HP group). We evaluated the alveolar gap reduction and the size of the maxilla between the two groups, obtaining intraoral maxillary impressions at birth, at 3 months and 1 year. The width of the alveolar gap in the β-TW plate group was significantly reduced compared with that in the HP group 1 month after the treatment (p pre-surgical orthodontic treatments. Copyright © 2013 British Association of Plastic, Reconstructive and Aesthetic Surgeons. Published by Elsevier Ltd. All rights reserved.

  6. Repair of single-strand breaks induced in the DNA of Proteus mirabilis by excision repair after UV-irradiation

    International Nuclear Information System (INIS)

    Stoerl, K.; Mund, C.

    1977-01-01

    Single-strand breaks have been produced in the DNA of P. mirabilis after UV-irradiation in dependence on the incident UV-doses. It has been found that there exists a discrepancy between the single-strand breaks estimated from sedimentation in alkaline sucrose gradients and the expected single-strand breaks approximated from measurements of dimer excision. The low number in incision breaks observed by sedimentation experiments is an indication that the cells are able to repair the excision-induced breaks as fast as they are formed. Toluenized cells have been used for investigation of the incision step independently of subsequent repair processes. In presence of NMN the appearance of more single-strand breaks in the DNA has been observed. Furthermore, the number of incision breaks in toluenized cells increased in presence of exogenous ATP. The completion of the excision repair process has been investigated by observing the rejoining of incision breaks. After irradiation with UV-doses higher than approximately 240 erg/mm 2 the number of single-strand breaks remaining unrepaired in the DNA increased. Studies of the influence of nutrition conditions on the repair process have shown approximately the same capacity for repair of single-strand breaks in growth medium as well as in buffer. Progress in the excision repair was also followed by investigation of the DNA synthesized at the template-DNA containing the pyrimidine dimers. In comparison with E. coli, P. mirabilis showed a somewhat lower efficiency for the repair of single-strand breaks during the excision repair. (author)

  7. Excess single-stranded DNA inhibits meiotic double-strand break repair.

    Directory of Open Access Journals (Sweden)

    Rebecca Johnson

    2007-11-01

    Full Text Available During meiosis, self-inflicted DNA double-strand breaks (DSBs are created by the protein Spo11 and repaired by homologous recombination leading to gene conversions and crossovers. Crossover formation is vital for the segregation of homologous chromosomes during the first meiotic division and requires the RecA orthologue, Dmc1. We analyzed repair during meiosis of site-specific DSBs created by another nuclease, VMA1-derived endonuclease (VDE, in cells lacking Dmc1 strand-exchange protein. Turnover and resection of the VDE-DSBs was assessed in two different reporter cassettes that can repair using flanking direct repeat sequences, thereby obviating the need for a Dmc1-dependent DNA strand invasion step. Access of the single-strand binding complex replication protein A, which is normally used in all modes of DSB repair, was checked in chromatin immunoprecipitation experiments, using antibody against Rfa1. Repair of the VDE-DSBs was severely inhibited in dmc1Delta cells, a defect that was associated with a reduction in the long tract resection required to initiate single-strand annealing between the flanking repeat sequences. Mutants that either reduce Spo11-DSB formation or abolish resection at Spo11-DSBs rescued the repair block. We also found that a replication protein A component, Rfa1, does not accumulate to expected levels at unrepaired single-stranded DNA (ssDNA in dmc1Delta cells. The requirement of Dmc1 for VDE-DSB repair using flanking repeats appears to be caused by the accumulation of large quantities of ssDNA that accumulate at Spo11-DSBs when Dmc1 is absent. We propose that these resected DSBs sequester both resection machinery and ssDNA binding proteins, which in wild-type cells would normally be recycled as Spo11-DSBs repair. The implication is that repair proteins are in limited supply, and this could reflect an underlying mechanism for regulating DSB repair in wild-type cells, providing protection from potentially harmful effects

  8. Mapping yeast origins of replication via single-stranded DNA detection.

    Science.gov (United States)

    Feng, Wenyi; Raghuraman, M K; Brewer, Bonita J

    2007-02-01

    Studies in th Saccharomyces cerevisiae have provided a framework for understanding how eukaryotic cells replicate their chromosomal DNA to ensure faithful transmission of genetic information to their daughter cells. In particular, S. cerevisiae is the first eukaryote to have its origins of replication mapped on a genomic scale, by three independent groups using three different microarray-based approaches. Here we describe a new technique of origin mapping via detection of single-stranded DNA in yeast. This method not only identified the majority of previously discovered origins, but also detected new ones. We have also shown that this technique can identify origins in Schizosaccharomyces pombe, illustrating the utility of this method for origin mapping in other eukaryotes.

  9. Single-strand breaks induced in Bacillus subtilis DNA by ultraviolet light: action spectrum and properties

    International Nuclear Information System (INIS)

    Peak, M.J.; Peak, J.G.

    1982-01-01

    The induction of single-strand breaks (alkali-labile bonds plus frank breaks) in the DNA of Bacillus subtilis irradiated in vivo by monochromatic UV light at wavelengths from 254 to 434nm was measured. The spectrum consists of a major far-UV (below 320nm) component and a minor near-UV shoulder. A mutant deficient in DNA polymerase I accumulates breaks caused by near-UV (above 320nm) wavelengths faster than the wild-type strain proficient in polymerase I. Measurable breaks in extracted DNA are induced at a higher frequency than those induced in vivo. Anoxia, glycerol, and diazobicyclo (2.2.2.) octane inhibit break formation in extracted DNA. Alkali-labile bonds induced by 365-nm UV radiation are largely (78%) covalent bond chain breaks, the remainder consists of true alkali-labile bonds, probably apurinic and apyrimidinic sites. (author)

  10. CdS nanowires formed by chemical synthesis using conjugated single-stranded DNA molecules

    Science.gov (United States)

    Sarangi, S. N.; Sahu, S. N.; Nozaki, S.

    2018-03-01

    CdS nanowires were successfully grown by chemical synthesis using two conjugated single-stranded (ss) DNA molecules, poly G (30) and poly C (30), as templates. During the early stage of the synthesis with the DNA molecules, the Cd 2+ interacts with Poly G and Poly C and produces the (Cd 2+)-Poly GC complex. As the growth proceeds, it results in nanowires. The structural analysis by grazing angle x-ray diffraction and transmission electron microscopy confirmed the zinc-blende CdS nanowires with the growth direction of . Although the nanowires are well surface-passivated with the DNA molecules, the photoluminescence quenching was caused by the electron transfer from the nanowires to the DNA molecules. The quenching can be used to detect and label the DNAs.

  11. The effects of radioprotective agents on the radiation-induced DNA single strand breaks

    International Nuclear Information System (INIS)

    Rhiu, Sung Ryul; Ko, Kyung Hwan; Jung, In Yong; Cho, Chul Ku; Kim, Tae Hwan; Park, Woo Wiun; Kim, Sung Ho; Ji, Young Hoon; Kim, Kyung Jung; Bang, Hio Chang; Jung, Young Suk; Choi, Moon Sik

    1992-04-01

    With the increased use of atomic energy in science, industry, medicine and public power production, the probability of nuclear accidents certainly appears to be on the increase. Therefore, early medical diagnosis and first-aid are needed urgently to establish an efficient treatment. We carried out the studies of radiation protector such as DDC, MEA, WR-2721 and variety of decontaminator with a view to establishing the protective measure and diagnostic standards for safety of worker and neighbors living around the radiation area in case of occurring the accidental contamination. In this experiment, we examined radiation-induced DNA single strand breaks as one of the study on molecular biology of the response of cells to radiation because an understanding of the radiation-induced damage in molecular level would add to our knowledge of radiation protection and treatment. (Author)

  12. Capillary Electrophoresis Single-Strand Conformational Polymorphisms as a Method to Differentiate Algal Species

    Directory of Open Access Journals (Sweden)

    Alice Jernigan

    2015-01-01

    Full Text Available Capillary electrophoresis single-strand conformational polymorphism (CE-SSCP was explored as a fast and inexpensive method to differentiate both prokaryotic (blue-green and eukaryotic (green and brown algae. A selection of two blue-green algae (Nostoc muscorum and Anabaena inaequalis, five green algae (Chlorella vulgaris, Oedogonium foveolatum, Mougeotia sp., Scenedesmus quadricauda, and Ulothrix fimbriata, and one brown algae (Ectocarpus sp. were examined and CE-SSCP electropherogram “fingerprints” were compared to each other for two variable regions of either the 16S or 18S rDNA gene. The electropherogram patterns were remarkably stable and consistent for each particular species. The patterns were unique to each species, although some common features were observed between the different types of algae. CE-SSCP could be a useful method for monitoring changes in an algae species over time as potential shifts in species occurred.

  13. Radioimmunoassay of single-stranded DNA antibodies for control of diagnosis and therapy

    Energy Technology Data Exchange (ETDEWEB)

    Meffert, H; Boehm, F; Soennichsen, N; Gens, J [Humboldt-Universitaet, Berlin (German Democratic Republic). Bereich Medizin (Charite)

    1980-10-01

    Several years experience in quantitative determination of single-stranded DNA antibodies is reported and the normal range as well as the diagnostic hit rate of the method is outlined. In the controls the mean DNA attachment rate was 1.5% and the upper normal range limit was 12.8%, the risk of erroneous rejection being 1%. The DNA binding rate was greater than 12.8% in 74.7% of untreated patients suffering from lupus erythematodes visceralis, in 47.6% of patients with circumscribed sclerodermia, in 14.4% of patients with progressive sclerodermia, and in 10.3% of those suffering from lupus erythematodes chronicus. The findings emphasize the importance of regulatory mechanisms of the immune system to the process of autosensitization.

  14. Radioimmunoassay of single-stranded DNA antibodies for control of diagnosis and therapy

    International Nuclear Information System (INIS)

    Meffert, H.; Boehm, F.; Soennichsen, N.; Gens, J.

    1980-01-01

    Several years experience in quantitative determination of single-stranded DNA antibodies is reported and the normal range as well as the diagnostic hit rate of the method is outlined. In the controls the mean DNA attachment rate was 1.5% and the upper normal range limit was 12.8%, the risk of erroneous rejection being 1%. The DNA binding rate was greater than 12.8% in 74.7% of untreated patients suffering from lupus erythematodes visceralis, in 47.6% of patients with circumscribed sclerodermia, in 14.4% of patients with progressive sclerodermia, and in 10.3% of those suffering from lupus erythematodes chronicus. The findings emphasize the importance of regulatory mechanisms of the immune system to the process of autosensitization

  15. Zinc(II) and the single-stranded DNA binding protein of bacteriophage T4

    International Nuclear Information System (INIS)

    Gauss, P.; Krassa, K.B.; McPheeters, D.S.; Nelson, M.A.; Gold, L.

    1987-01-01

    The DNA binding domain of the gene 32 protein of the bacteriophage T4 contains a single zinc-finger sequence. The gene 32 protein is an extensively studied member of a class of proteins that bind relatively nonspecifically to single-stranded DNA. The authors have sequenced and characterized mutations in gene 32 whose defective proteins are activated by increasing the Zn(II) concentration in the growth medium. The results identify a role for the gene 32 protein in activation of T4 late transcription. Several eukaryotic proteins with zinc fingers participate in activation of transcription, and the gene 32 protein of T4 should provide a simple, well-characterized system in which genetics can be utilized to study the role of a zinc finger in nucleic acid binding and gene expression

  16. Bacterial single-stranded DNA-binding proteins are phosphorylated on tyrosine

    DEFF Research Database (Denmark)

    Mijakovic, Ivan; Petranovic, Dina; Macek, B

    2006-01-01

    for phosphotyrosine-containing proteins in Streptomyces griseus by immunoaffinity chromatography identified bacterial SSBs as a novel target of bacterial tyrosine kinases. Since genes encoding protein-tyrosine kinases (PTKs) have not been recognized in streptomycetes, and SSBs from Streptomyces coelicolor (Sc......SSB) and Bacillus subtilis (BsSSB) share 38.7% identity, we used a B.subtilis protein-tyrosine kinase YwqD to phosphorylate two cognate SSBs (BsSSB and YwpH) in vitro. We demonstrate that in vivo phosphorylation of B.subtilis SSB occurs on tyrosine residue 82, and this reaction is affected antagonistically...... by kinase YwqD and phosphatase YwqE. Phosphorylation of B.subtilis SSB increased binding almost 200-fold to single-stranded DNA in vitro. Tyrosine phosphorylation of B.subtilis, S.coelicolor and Escherichia coli SSBs occured while they were expressed in E.coli, indicating that tyrosine phosphorylation...

  17. Towards quantitative viromics for both double-stranded and single-stranded DNA viruses

    Directory of Open Access Journals (Sweden)

    Simon Roux

    2016-12-01

    Full Text Available Background Viruses strongly influence microbial population dynamics and ecosystem functions. However, our ability to quantitatively evaluate those viral impacts is limited to the few cultivated viruses and double-stranded DNA (dsDNA viral genomes captured in quantitative viral metagenomes (viromes. This leaves the ecology of non-dsDNA viruses nearly unknown, including single-stranded DNA (ssDNA viruses that have been frequently observed in viromes, but not quantified due to amplification biases in sequencing library preparations (Multiple Displacement Amplification, Linker Amplification or Tagmentation. Methods Here we designed mock viral communities including both ssDNA and dsDNA viruses to evaluate the capability of a sequencing library preparation approach including an Adaptase step prior to Linker Amplification for quantitative amplification of both dsDNA and ssDNA templates. We then surveyed aquatic samples to provide first estimates of the abundance of ssDNA viruses. Results Mock community experiments confirmed the biased nature of existing library preparation methods for ssDNA templates (either largely enriched or selected against and showed that the protocol using Adaptase plus Linker Amplification yielded viromes that were ±1.8-fold quantitative for ssDNA and dsDNA viruses. Application of this protocol to community virus DNA from three freshwater and three marine samples revealed that ssDNA viruses as a whole represent only a minor fraction (<5% of DNA virus communities, though individual ssDNA genomes, both eukaryote-infecting Circular Rep-Encoding Single-Stranded DNA (CRESS-DNA viruses and bacteriophages from the Microviridae family, can be among the most abundant viral genomes in a sample. Discussion Together these findings provide empirical data for a new virome library preparation protocol, and a first estimate of ssDNA virus abundance in aquatic systems.

  18. Accurate quantification of microRNA via single strand displacement reaction on DNA origami motif.

    Directory of Open Access Journals (Sweden)

    Jie Zhu

    Full Text Available DNA origami is an emerging technology that assembles hundreds of staple strands and one single-strand DNA into certain nanopattern. It has been widely used in various fields including detection of biological molecules such as DNA, RNA and proteins. MicroRNAs (miRNAs play important roles in post-transcriptional gene repression as well as many other biological processes such as cell growth and differentiation. Alterations of miRNAs' expression contribute to many human diseases. However, it is still a challenge to quantitatively detect miRNAs by origami technology. In this study, we developed a novel approach based on streptavidin and quantum dots binding complex (STV-QDs labeled single strand displacement reaction on DNA origami to quantitatively detect the concentration of miRNAs. We illustrated a linear relationship between the concentration of an exemplary miRNA as miRNA-133 and the STV-QDs hybridization efficiency; the results demonstrated that it is an accurate nano-scale miRNA quantifier motif. In addition, both symmetrical rectangular motif and asymmetrical China-map motif were tested. With significant linearity in both motifs, our experiments suggested that DNA Origami motif with arbitrary shape can be utilized in this method. Since this DNA origami-based method we developed owns the unique advantages of simple, time-and-material-saving, potentially multi-targets testing in one motif and relatively accurate for certain impurity samples as counted directly by atomic force microscopy rather than fluorescence signal detection, it may be widely used in quantification of miRNAs.

  19. Accurate Quantification of microRNA via Single Strand Displacement Reaction on DNA Origami Motif

    Science.gov (United States)

    Lou, Jingyu; Li, Weidong; Li, Sheng; Zhu, Hongxin; Yang, Lun; Zhang, Aiping; He, Lin; Li, Can

    2013-01-01

    DNA origami is an emerging technology that assembles hundreds of staple strands and one single-strand DNA into certain nanopattern. It has been widely used in various fields including detection of biological molecules such as DNA, RNA and proteins. MicroRNAs (miRNAs) play important roles in post-transcriptional gene repression as well as many other biological processes such as cell growth and differentiation. Alterations of miRNAs' expression contribute to many human diseases. However, it is still a challenge to quantitatively detect miRNAs by origami technology. In this study, we developed a novel approach based on streptavidin and quantum dots binding complex (STV-QDs) labeled single strand displacement reaction on DNA origami to quantitatively detect the concentration of miRNAs. We illustrated a linear relationship between the concentration of an exemplary miRNA as miRNA-133 and the STV-QDs hybridization efficiency; the results demonstrated that it is an accurate nano-scale miRNA quantifier motif. In addition, both symmetrical rectangular motif and asymmetrical China-map motif were tested. With significant linearity in both motifs, our experiments suggested that DNA Origami motif with arbitrary shape can be utilized in this method. Since this DNA origami-based method we developed owns the unique advantages of simple, time-and-material-saving, potentially multi-targets testing in one motif and relatively accurate for certain impurity samples as counted directly by atomic force microscopy rather than fluorescence signal detection, it may be widely used in quantification of miRNAs. PMID:23990889

  20. Accurate quantification of microRNA via single strand displacement reaction on DNA origami motif.

    Science.gov (United States)

    Zhu, Jie; Feng, Xiaolu; Lou, Jingyu; Li, Weidong; Li, Sheng; Zhu, Hongxin; Yang, Lun; Zhang, Aiping; He, Lin; Li, Can

    2013-01-01

    DNA origami is an emerging technology that assembles hundreds of staple strands and one single-strand DNA into certain nanopattern. It has been widely used in various fields including detection of biological molecules such as DNA, RNA and proteins. MicroRNAs (miRNAs) play important roles in post-transcriptional gene repression as well as many other biological processes such as cell growth and differentiation. Alterations of miRNAs' expression contribute to many human diseases. However, it is still a challenge to quantitatively detect miRNAs by origami technology. In this study, we developed a novel approach based on streptavidin and quantum dots binding complex (STV-QDs) labeled single strand displacement reaction on DNA origami to quantitatively detect the concentration of miRNAs. We illustrated a linear relationship between the concentration of an exemplary miRNA as miRNA-133 and the STV-QDs hybridization efficiency; the results demonstrated that it is an accurate nano-scale miRNA quantifier motif. In addition, both symmetrical rectangular motif and asymmetrical China-map motif were tested. With significant linearity in both motifs, our experiments suggested that DNA Origami motif with arbitrary shape can be utilized in this method. Since this DNA origami-based method we developed owns the unique advantages of simple, time-and-material-saving, potentially multi-targets testing in one motif and relatively accurate for certain impurity samples as counted directly by atomic force microscopy rather than fluorescence signal detection, it may be widely used in quantification of miRNAs.

  1. Kinetics of end-to-end collision in short single-stranded nucleic acids.

    Science.gov (United States)

    Wang, Xiaojuan; Nau, Werner M

    2004-01-28

    A novel fluorescence-based method, which entails contact quenching of the long-lived fluorescent state of 2,3-diazabicyclo[2.2.2]-oct-2-ene (DBO), was employed to measure the kinetics of end-to-end collision in short single-stranded oligodeoxyribonucleotides of the type 5'-DBO-(X)n-dG with X = dA, dC, dT, or dU and n = 2 or 4. The fluorophore was covalently attached to the 5' end and dG was introduced as an efficient intrinsic quencher at the 3' terminus. The end-to-end collision rates, which can be directly related to the efficiency of intramolecular fluorescence quenching, ranged from 0.1 to 9.0 x 10(6) s(-1). They were strongly dependent on the strand length, the base sequence, as well as the temperature. Oligonucleotides containing dA in the backbone displayed much slower collision rates and significantly higher positive activation energies than strands composed of pyrimidine bases, suggesting a higher intrinsic rigidity of oligoadenylate. Comparison of the measured collision rates in short single-stranded oligodeoxyribonucleotides with the previously reported kinetics of hairpin formation indicates that the intramolecular collision is significantly faster than the nucleation step of hairpin closing. This is consistent with the configurational diffusion model suggested by Ansari et al. (Ansari, A.; Kuznetsov, S. V.; Shen, Y. Proc.Natl. Acad. Sci. USA 2001, 98, 7771-7776), in which the formation of misfolded loops is thought to slow hairpin formation.

  2. Yield of single-strand breaks in the DNA of E.coli 10 msec after irradiation

    International Nuclear Information System (INIS)

    Fox, R.A.; Fielden, E.M.; Sapora, O.

    1976-01-01

    The rapid mixing of 0.3M alkali with a suspension of E.coli B/r 6 +- 3 and 144 +- 3 msec after irradiation with electrons (4.3 MeV, 0 to 50 krad) has been used to make a comparison of the yields of single strand breaks in the presence and absence of oxygen. No significant difference was observed between the numbers of single strand breaks appearing at 6 and 144 msec after irradiation. Assuming that mixing with alkali inactivates the cellular repair enzymes within several milliseconds, these results indicate that enzymic repair does not operate within this time scale. It seems probable that radiation chemical processes are responsible for the initial oxygen effect on single strand breaks. (U.K.)

  3. Strand Displacement by DNA Polymerase III Occurs through a τ-ψ-χ Link to Single-stranded DNA-binding Protein Coating the Lagging Strand Template*

    OpenAIRE

    Yuan, Quan; McHenry, Charles S.

    2009-01-01

    In addition to the well characterized processive replication reaction catalyzed by the DNA polymerase III holoenzyme on single-stranded DNA templates, the enzyme possesses an intrinsic strand displacement activity on flapped templates. The strand displacement activity is distinguished from the single-stranded DNA-templated reaction by a high dependence upon single-stranded DNA binding protein and an inability of γ-complex to support the reaction in the absence of τ. However, if γ-complex is p...

  4. Mechanical properties of NiTi and CuNiTi wires used in orthodontic treatment. Part 2: Microscopic surface appraisal and metallurgical characteristics

    Directory of Open Access Journals (Sweden)

    Marco Abdo Gravina

    2014-01-01

    Full Text Available OBJECTIVE: This research aimed at comparing the qualitative chemical compositions and the surface morphology of fracture regions of eight types of Nickel (Ni Titanium (Ti conventional wires, superelastic and heat-activated (GAC, TP, Ormco, Masel, Morelli and Unitek, to the wires with addition of copper (CuNiTi 27oC and 35oC, Ormco after traction test. METHODS: The analyses were performed in a scanning electronic microscope (JEOL, model JSM-5800 LV with EDS system of microanalysis (energy dispersive spectroscopy. RESULTS : The results showed that NiTi wires presented Ni and Ti as the main elements of the alloy with minimum differences in their composition. The CuNiTi wires, however, presented Ni and Ti with a significant percentage of copper (Cu. As for surface morphology, the wires that presented the lowest wire-surface roughness were the superelastic ones by Masel and Morelli, while those that presented the greatest wire-surface roughness were the CuNiTi 27oC and 35oC ones by Ormco, due to presence of microcavity formed as a result of pulling out some particles, possibly of NiTi. 4 The fracture surfaces presented characteristics of ductile fracture, with presence of microcavities. The superelastic wires by GAC and the CuNiTi 27oC and the heat-activated ones by Unitek presented the smallest microcavities and the lowest wire-surface roughness with regard to fracture, while the CuNiTi 35oC wires presented inadequate wire-surface roughness in the fracture region. CONCLUSION: CuNiTi 35oC wires did not present better morphologic characteristics in comparison to the other wires with regard to surfaces and fracture region.

  5. Single-strand conformation polymorphism analysis of ribosomal DNA for detection of Phytophthora ramorum directly from plant tissues

    Science.gov (United States)

    Ping Kong; Patricia A. Richardson; Chuanxue Hong; Thomas L. Kubisiak

    2006-01-01

    At the first Sudden Oak Death Science Symposium, we reported on the use of a single strand conformation polymorphism (SSCP) analysis for rapid identification of Phytophthora ramorum in culture. We have since assessed and improved the fingerprinting technique for detecting this pathogen directly from plant tissues. The improved SSCP protocol uses a...

  6. Novel Single-Stranded DNA Virus Genomes Recovered from Chimpanzee Feces Sampled from the Mambilla Plateau in Nigeria

    Science.gov (United States)

    Walters, Matthew; Bawuro, Musa; Christopher, Alfred; Knight, Alexander; Kraberger, Simona; Stainton, Daisy; Chapman, Hazel

    2017-01-01

    ABSTRACT Metagenomic approaches are rapidly expanding our knowledge of the diversity of viruses. In the fecal matter of Nigerian chimpanzees we recovered three gokushovirus genomes, one circular replication-associated protein encoding single-stranded DNA virus (CRESS), and a CRESS DNA molecule. PMID:28254982

  7. Interaction with Single-stranded DNA-binding Protein Stimulates Escherichia coli Ribonuclease HI Enzymatic Activity.

    Science.gov (United States)

    Petzold, Christine; Marceau, Aimee H; Miller, Katherine H; Marqusee, Susan; Keck, James L

    2015-06-05

    Single-stranded (ss) DNA-binding proteins (SSBs) bind and protect ssDNA intermediates formed during replication, recombination, and repair reactions. SSBs also directly interact with many different genome maintenance proteins to stimulate their enzymatic activities and/or mediate their proper cellular localization. We have identified an interaction formed between Escherichia coli SSB and ribonuclease HI (RNase HI), an enzyme that hydrolyzes RNA in RNA/DNA hybrids. The RNase HI·SSB complex forms by RNase HI binding the intrinsically disordered C terminus of SSB (SSB-Ct), a mode of interaction that is shared among all SSB interaction partners examined to date. Residues that comprise the SSB-Ct binding site are conserved among bacterial RNase HI enzymes, suggesting that RNase HI·SSB complexes are present in many bacterial species and that retaining the interaction is important for its cellular function. A steady-state kinetic analysis shows that interaction with SSB stimulates RNase HI activity by lowering the reaction Km. SSB or RNase HI protein variants that disrupt complex formation nullify this effect. Collectively our findings identify a direct RNase HI/SSB interaction that could play a role in targeting RNase HI activity to RNA/DNA hybrid substrates within the genome. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  8. Interaction with Single-stranded DNA-binding Protein Stimulates Escherichia coli Ribonuclease HI Enzymatic Activity*

    Science.gov (United States)

    Petzold, Christine; Marceau, Aimee H.; Miller, Katherine H.; Marqusee, Susan; Keck, James L.

    2015-01-01

    Single-stranded (ss) DNA-binding proteins (SSBs) bind and protect ssDNA intermediates formed during replication, recombination, and repair reactions. SSBs also directly interact with many different genome maintenance proteins to stimulate their enzymatic activities and/or mediate their proper cellular localization. We have identified an interaction formed between Escherichia coli SSB and ribonuclease HI (RNase HI), an enzyme that hydrolyzes RNA in RNA/DNA hybrids. The RNase HI·SSB complex forms by RNase HI binding the intrinsically disordered C terminus of SSB (SSB-Ct), a mode of interaction that is shared among all SSB interaction partners examined to date. Residues that comprise the SSB-Ct binding site are conserved among bacterial RNase HI enzymes, suggesting that RNase HI·SSB complexes are present in many bacterial species and that retaining the interaction is important for its cellular function. A steady-state kinetic analysis shows that interaction with SSB stimulates RNase HI activity by lowering the reaction Km. SSB or RNase HI protein variants that disrupt complex formation nullify this effect. Collectively our findings identify a direct RNase HI/SSB interaction that could play a role in targeting RNase HI activity to RNA/DNA hybrid substrates within the genome. PMID:25903123

  9. Cdc45-induced loading of human RPA onto single-stranded DNA.

    Science.gov (United States)

    Szambowska, Anna; Tessmer, Ingrid; Prus, Piotr; Schlott, Bernhard; Pospiech, Helmut; Grosse, Frank

    2017-04-07

    Cell division cycle protein 45 (Cdc45) is an essential component of the eukaryotic replicative DNA helicase. We found that human Cdc45 forms a complex with the single-stranded DNA (ssDNA) binding protein RPA. Moreover, it actively loads RPA onto nascent ssDNA. Pull-down assays and surface plasmon resonance studies revealed that Cdc45-bound RPA complexed with ssDNA in the 8-10 nucleotide binding mode, but dissociated when RPA covered a 30-mer. Real-time analysis of RPA-ssDNA binding demonstrated that Cdc45 catalytically loaded RPA onto ssDNA. This placement reaction required physical contacts of Cdc45 with the RPA70A subdomain. Our results imply that Cdc45 controlled stabilization of the 8-nt RPA binding mode, the subsequent RPA transition into 30-mer mode and facilitated an ordered binding to ssDNA. We propose that a Cdc45-mediated loading guarantees a seamless deposition of RPA on newly emerging ssDNA at the nascent replication fork. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  10. Leishmania replication protein A-1 binds in vivo single-stranded telomeric DNA

    International Nuclear Information System (INIS)

    Neto, J.L. Siqueira; Lira, C.B.B.; Giardini, M.A.; Khater, L.; Perez, A.M.; Peroni, L.A.; Reis, J.R.R. dos; Freitas-Junior, L.H.; Ramos, C.H.I.; Cano, M.I.N.

    2007-01-01

    Replication protein A (RPA) is a highly conserved heterotrimeric single-stranded DNA-binding protein involved in different events of DNA metabolism. In yeast, subunits 1 (RPA-1) and 2 (RPA-2) work also as telomerase recruiters and, in humans, the complex unfolds G-quartet structures formed by the 3' G-rich telomeric strand. In most eukaryotes, RPA-1 and RPA-2 bind DNA using multiple OB fold domains. In trypanosomatids, including Leishmania, RPA-1 has a canonical OB fold and a truncated RFA-1 structural domain. In Leishmania amazonensis, RPA-1 alone can form a complex in vitro with the telomeric G-rich strand. In this work, we show that LaRPA-1 is a nuclear protein that associates in vivo with Leishmania telomeres. We mapped the boundaries of the OB fold DNA-binding domain using deletion mutants. Since Leishmania and other trypanosomatids lack homologues of known telomere end binding proteins, our results raise questions about the function of RPA-1 in parasite telomeres

  11. Viral interference with DNA repair by targeting of the single-stranded DNA binding protein RPA.

    Science.gov (United States)

    Banerjee, Pubali; DeJesus, Rowena; Gjoerup, Ole; Schaffhausen, Brian S

    2013-10-01

    Correct repair of damaged DNA is critical for genomic integrity. Deficiencies in DNA repair are linked with human cancer. Here we report a novel mechanism by which a virus manipulates DNA damage responses. Infection with murine polyomavirus sensitizes cells to DNA damage by UV and etoposide. Polyomavirus large T antigen (LT) alone is sufficient to sensitize cells 100 fold to UV and other kinds of DNA damage. This results in activated stress responses and apoptosis. Genetic analysis shows that LT sensitizes via the binding of its origin-binding domain (OBD) to the single-stranded DNA binding protein replication protein A (RPA). Overexpression of RPA protects cells expressing OBD from damage, and knockdown of RPA mimics the LT phenotype. LT prevents recruitment of RPA to nuclear foci after DNA damage. This leads to failure to recruit repair proteins such as Rad51 or Rad9, explaining why LT prevents repair of double strand DNA breaks by homologous recombination. A targeted intervention directed at RPA based on this viral mechanism could be useful in circumventing the resistance of cancer cells to therapy.

  12. RPA Stabilization of Single-Stranded DNA Is Critical for Break-Induced Replication.

    Science.gov (United States)

    Ruff, Patrick; Donnianni, Roberto A; Glancy, Eleanor; Oh, Julyun; Symington, Lorraine S

    2016-12-20

    DNA double-strand breaks (DSBs) are cytotoxic lesions that must be accurately repaired to maintain genome stability. Replication protein A (RPA) plays an important role in homology-dependent repair of DSBs by protecting the single-stranded DNA (ssDNA) intermediates formed by end resection and by facilitating Rad51 loading. We found that hypomorphic mutants of RFA1 that support intra-chromosomal homologous recombination are profoundly defective for repair processes involving long tracts of DNA synthesis, in particular break-induced replication (BIR). The BIR defects of the rfa1 mutants could be partially suppressed by eliminating the Sgs1-Dna2 resection pathway, suggesting that Dna2 nuclease attacks the ssDNA formed during end resection when not fully protected by RPA. Overexpression of Rad51 was also found to suppress the rfa1 BIR defects. We suggest that Rad51 binding to the ssDNA formed by excessive end resection and during D-loop migration can partially compensate for dysfunctional RPA. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  13. Reconstitution of RPA-covered single-stranded DNA-activated ATR-Chk1 signaling.

    Science.gov (United States)

    Choi, Jun-Hyuk; Lindsey-Boltz, Laura A; Kemp, Michael; Mason, Aaron C; Wold, Marc S; Sancar, Aziz

    2010-08-03

    ATR kinase is a critical upstream regulator of the checkpoint response to various forms of DNA damage. Previous studies have shown that ATR is recruited via its binding partner ATR-interacting protein (ATRIP) to replication protein A (RPA)-covered single-stranded DNA (RPA-ssDNA) generated at sites of DNA damage where ATR is then activated by TopBP1 to phosphorylate downstream targets including the Chk1 signal transducing kinase. However, this critical feature of the human ATR-initiated DNA damage checkpoint signaling has not been demonstrated in a defined system. Here we describe an in vitro checkpoint system in which RPA-ssDNA and TopBP1 are essential for phosphorylation of Chk1 by the purified ATR-ATRIP complex. Checkpoint defective RPA mutants fail to activate ATR kinase in this system, supporting the conclusion that this system is a faithful representation of the in vivo reaction. Interestingly, we find that an alternative form of RPA (aRPA), which does not support DNA replication, can substitute for the checkpoint function of RPA in vitro, thus revealing a potential role for aRPA in the activation of ATR kinase. We also find that TopBP1 is recruited to RPA-ssDNA in a manner dependent on ATRIP and that the N terminus of TopBP1 is required for efficient recruitment and activation of ATR kinase.

  14. Characterization of a novel single-stranded RNA mycovirus in pleurotus ostreatus

    International Nuclear Information System (INIS)

    Yu, Hyun Jae; Lim, Dongbin; Lee, Hyun-Sook

    2003-01-01

    A mycovirus, named oyster mushroom spherical virus (OMSV), was isolated from cultivated oyster mushrooms with a severe epidemic of oyster mushroom Die-back disease. OMSV was a 27-nm spherical virus encapsidating a single-stranded RNA (ssRNA) of 5.784 kb with a coat protein of approximately 28.5 kDa. The nucleotide sequence of the virus revealed that its genomic RNA was positive strand, containing 5784 bases with seven open reading frames (ORF). ORF1 had the motifs of RNA-dependent RNA polymerases (RdRp) and helicase. ORF2 encoded a coat protein. ORF3 to 7 could encode putative polypeptides of approximately 12, 12.5, 21, 14.5, and 23 kDa, respectively, but none of them showed significant similarity to any other known polypeptides. The 5' end of the viral RNA was uncapped and the 3' end was polyadenylated with 74 bases. Genomic structure and organization and the derived amino acid sequence of RdRp and helicase domain were similar to those of tymoviruses, a plant virus group

  15. Single-stranded DNA cleavage by divergent CRISPR-Cas9 enzymes

    Science.gov (United States)

    Ma, Enbo; Harrington, Lucas B.; O’Connell, Mitchell R.; Zhou, Kaihong; Doudna, Jennifer A.

    2015-01-01

    Summary Double-stranded DNA (dsDNA) cleavage by Cas9 is a hallmark of type II CRISPR-Cas immune systems. Cas9–guide RNA complexes recognize 20-base-pair sequences in DNA and generate a site-specific double-strand break, a robust activity harnessed for genome editing. DNA recognition by all studied Cas9 enzymes requires a protospacer adjacent motif (PAM) next to the target site. We show that Cas9 enzymes from evolutionarily divergent bacteria can recognize and cleave single-stranded DNA (ssDNA) by an RNA-guided, PAM-independent recognition mechanism. Comparative analysis shows that in contrast to the type II-A S. pyogenes Cas9 that is widely used for genome engineering, the smaller type II-C Cas9 proteins have limited dsDNA binding and unwinding activity and promiscuous guide-RNA specificity. These results indicate that inefficiency of type II-C Cas9 enzymes for genome editing results from a limited ability to cleave dsDNA, and suggest that ssDNA cleavage was an ancestral function of the Cas9 enzyme family. PMID:26545076

  16. Fragment-based modelling of single stranded RNA bound to RNA recognition motif containing proteins

    Science.gov (United States)

    de Beauchene, Isaure Chauvot; de Vries, Sjoerd J.; Zacharias, Martin

    2016-01-01

    Abstract Protein-RNA complexes are important for many biological processes. However, structural modeling of such complexes is hampered by the high flexibility of RNA. Particularly challenging is the docking of single-stranded RNA (ssRNA). We have developed a fragment-based approach to model the structure of ssRNA bound to a protein, based on only the protein structure, the RNA sequence and conserved contacts. The conformational diversity of each RNA fragment is sampled by an exhaustive library of trinucleotides extracted from all known experimental protein–RNA complexes. The method was applied to ssRNA with up to 12 nucleotides which bind to dimers of the RNA recognition motifs (RRMs), a highly abundant eukaryotic RNA-binding domain. The fragment based docking allows a precise de novo atomic modeling of protein-bound ssRNA chains. On a benchmark of seven experimental ssRNA–RRM complexes, near-native models (with a mean heavy-atom deviation of <3 Å from experiment) were generated for six out of seven bound RNA chains, and even more precise models (deviation < 2 Å) were obtained for five out of seven cases, a significant improvement compared to the state of the art. The method is not restricted to RRMs but was also successfully applied to Pumilio RNA binding proteins. PMID:27131381

  17. Packaging signals in single-stranded RNA viruses: nature's alternative to a purely electrostatic assembly mechanism.

    Science.gov (United States)

    Stockley, Peter G; Twarock, Reidun; Bakker, Saskia E; Barker, Amy M; Borodavka, Alexander; Dykeman, Eric; Ford, Robert J; Pearson, Arwen R; Phillips, Simon E V; Ranson, Neil A; Tuma, Roman

    2013-03-01

    The formation of a protective protein container is an essential step in the life-cycle of most viruses. In the case of single-stranded (ss)RNA viruses, this step occurs in parallel with genome packaging in a co-assembly process. Previously, it had been thought that this process can be explained entirely by electrostatics. Inspired by recent single-molecule fluorescence experiments that recapitulate the RNA packaging specificity seen in vivo for two model viruses, we present an alternative theory, which recognizes the important cooperative roles played by RNA-coat protein interactions, at sites we have termed packaging signals. The hypothesis is that multiple copies of packaging signals, repeated according to capsid symmetry, aid formation of the required capsid protein conformers at defined positions, resulting in significantly enhanced assembly efficiency. The precise mechanistic roles of packaging signal interactions may vary between viruses, as we have demonstrated for MS2 and STNV. We quantify the impact of packaging signals on capsid assembly efficiency using a dodecahedral model system, showing that heterogeneous affinity distributions of packaging signals for capsid protein out-compete those of homogeneous affinities. These insights pave the way to a new anti-viral therapy, reducing capsid assembly efficiency by targeting of the vital roles of the packaging signals, and opens up new avenues for the efficient construction of protein nanocontainers in bionanotechnology.

  18. Kinetics of repair of DNA single-strand breaks in cultured mammalian cells

    International Nuclear Information System (INIS)

    Vexler, F.B.; Eidus, L.Kh.; Vexler, A.M.

    1984-01-01

    Postirradiation treatment of Chinese hamster cells with cysteamine (MEA), caffeine-benzoate (CB) and caffeine sharply inhibits the repair of DNA single-strand breaks in the first five minutes. This inhibition is reversible since removing of the agent leads immediately to the resumption of the repair. The rate of the repair is decreased with prolongation of treatment and increasing concentration of the modifying agent. The efficiency of the substances studied depends not only on their concentration in the medium. For MEA and CB, which are weak electrolytes, it is also pH-dependent. This is explained by the theory of dissociation of weak electrolytes and their distribution between the cell and medium. It is shown that intracellular concentration of the substances is the most important factor determining their efficiency. All the three substances exert practically the same effect when compared at equal intracellular concentration. The above presented data serve as evidence for the existence of an unspecific mechanism of the effect of the substances studied. (author)

  19. Precise gene modification mediated by TALEN and single-stranded oligodeoxynucleotides in human cells.

    Directory of Open Access Journals (Sweden)

    Xiaoling Wang

    Full Text Available The development of human embryonic stem cells (ESCs and induced pluripotent stem cells (iPSCs facilitates in vitro studies of human disease mechanisms, speeds up the process of drug screening, and raises the feasibility of using cell replacement therapy in clinics. However, the study of genotype-phenotype relationships in ESCs or iPSCs is hampered by the low efficiency of site-specific gene editing. Transcription activator-like effector nucleases (TALENs spurred interest due to the ease of assembly, high efficiency and faithful gene targeting. In this study, we optimized the TALEN design to maximize its genomic cutting efficiency. We showed that using optimized TALENs in conjunction with single-strand oligodeoxynucleotide (ssODN allowed efficient gene editing in human cells. Gene mutations and gene deletions for up to 7.8 kb can be accomplished at high efficiencies. We established human tumor cell lines and H9 ESC lines with homozygous deletion of the microRNA-21 (miR-21 gene and miR-9-2 gene. These cell lines provide a robust platform to dissect the roles these genes play during cell differentiation and tumorigenesis. We also observed that the endogenous homologous chromosome can serve as a donor template for gene editing. Overall, our studies demonstrate the versatility of using ssODN and TALEN to establish genetically modified cells for research and therapeutic application.

  20. Selection and characterization of single stranded DNA aptamers recognizing fumonisin B1

    International Nuclear Information System (INIS)

    Chen, Xiujuan; Huang, Yukun; Duan, Nuo; Wu, Shijia; Xia, Yu; Ma, Xiaoyuan; Ding, Zhansheng; Wang, Zhouping; Zhu, Changqing; Jiang, Yuan

    2014-01-01

    We present an improved method for the selection of single-stranded DNA aptamers that can recognize fumonisin B 1 (FB 1 ). FB 1 is a carcinogenic mycotoxin mainly found in corn and corn-based food products worldwide, posing a global threat to feed and food safety. Selection was based on the mag-SELEX (magnetic bead systematic evolution of ligands by exponential enrichment) technology modified by adopting free analogs of targets rather than immobilized targets for counter selections. Firstly, aptamer candidates for FB 1 were selected from an 80 nt random DNA library after 13 rounds of selection. Next, binding assays were performed for affinity evaluation, and circular dichroism spectroscopy was used to investigate their conformation. A high-affinity aptamer designated as F10 (with a dissociation constant of 62 ± 5 nM) was identified and tested for its specificity by competitive binding assays. The results demonstrate that this improved mag-SELEX technology facilitates aptamer screening because it avoids the tedious immobilization of counter-selection molecules on magnetic beads. The aptamers obtained by this technique open new possibilities for the detection of FB 1 via aptasensors. (author)

  1. Selection and characterization of single stranded DNA aptamers recognizing fumonisin B{sub 1}

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Xiujuan; Huang, Yukun; Duan, Nuo; Wu, Shijia; Xia, Yu; Ma, Xiaoyuan; Ding, Zhansheng; Wang, Zhouping [State Key Laboratory of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, School of Food Science and Technology, Jiangnan University, Wuxi, 214122 (China); Zhu, Changqing; Jiang, Yuan [Animal, Plant and Food Inspection Centre, Jiangsu Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001 (China)

    2014-08-01

    We present an improved method for the selection of single-stranded DNA aptamers that can recognize fumonisin B{sub 1} (FB{sub 1}). FB{sub 1} is a carcinogenic mycotoxin mainly found in corn and corn-based food products worldwide, posing a global threat to feed and food safety. Selection was based on the mag-SELEX (magnetic bead systematic evolution of ligands by exponential enrichment) technology modified by adopting free analogs of targets rather than immobilized targets for counter selections. Firstly, aptamer candidates for FB{sub 1} were selected from an 80 nt random DNA library after 13 rounds of selection. Next, binding assays were performed for affinity evaluation, and circular dichroism spectroscopy was used to investigate their conformation. A high-affinity aptamer designated as F10 (with a dissociation constant of 62 ± 5 nM) was identified and tested for its specificity by competitive binding assays. The results demonstrate that this improved mag-SELEX technology facilitates aptamer screening because it avoids the tedious immobilization of counter-selection molecules on magnetic beads. The aptamers obtained by this technique open new possibilities for the detection of FB{sub 1} via aptasensors. (author)

  2. Molecular dynamics simulation of a DNA containing a single strand break

    Energy Technology Data Exchange (ETDEWEB)

    Yamaguchi, H.; Siebers, G.; Furukawa, A.; Otagiri, N.; Osman, R

    2002-07-01

    Molecular dynamics simulations were performed for a dodecamer DNA containing a single strand break (SSB), which has been represented by a 3'-OH deoxyribose and 5'-OH phosphate in the middle of the strand. Molecular force field parameters of the 5'-OH phosphate region were determined from an ab initio calculation at the HF/6-31G level using the program package GAMESS. The DNA was placed in a periodic boundary box with water molecules and Na+ counter-ions to produce a neutralised system. After minimisation, the system was heated to 300 K, equilibrated and a production run at constant NTP was executed for 1 ns using AMBER 4.1. Snapshots of the SSB-containing DNA and a detailed analysis of the equilibriated average structure revealed surprisingly small conformational changes compared to normal DNA. However, dynamic properties calculated using the essential dynamics method showed some features that may be important for the recognition of this damage by repair enzymes. (author)

  3. A conserved MCM single-stranded DNA binding element is essential for replication initiation.

    Science.gov (United States)

    Froelich, Clifford A; Kang, Sukhyun; Epling, Leslie B; Bell, Stephen P; Enemark, Eric J

    2014-04-01

    The ring-shaped MCM helicase is essential to all phases of DNA replication. The complex loads at replication origins as an inactive double-hexamer encircling duplex DNA. Helicase activation converts this species to two active single hexamers that encircle single-stranded DNA (ssDNA). The molecular details of MCM DNA interactions during these events are unknown. We determined the crystal structure of the Pyrococcus furiosus MCM N-terminal domain hexamer bound to ssDNA and define a conserved MCM-ssDNA binding motif (MSSB). Intriguingly, ssDNA binds the MCM ring interior perpendicular to the central channel with defined polarity. In eukaryotes, the MSSB is conserved in several Mcm2-7 subunits, and MSSB mutant combinations in S. cerevisiae Mcm2-7 are not viable. Mutant Mcm2-7 complexes assemble and are recruited to replication origins, but are defective in helicase loading and activation. Our findings identify an important MCM-ssDNA interaction and suggest it functions during helicase activation to select the strand for translocation. DOI: http://dx.doi.org/10.7554/eLife.01993.001.

  4. Systematic Identification of Determinants for Single-Strand Annealing-Mediated Deletion Formation in Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Maia Segura-Wang

    2017-10-01

    Full Text Available To ensure genomic integrity, living organisms have evolved diverse molecular processes for sensing and repairing damaged DNA. If improperly repaired, DNA damage can give rise to different types of mutations, an important class of which are genomic structural variants (SVs. In spite of their importance for phenotypic variation and genome evolution, potential contributors to SV formation in Saccharomyces cerevisiae (budding yeast, a highly tractable model organism, are not fully recognized. Here, we developed and applied a genome-wide assay to identify yeast gene knockout mutants associated with de novo deletion formation, in particular single-strand annealing (SSA-mediated deletion formation, in a systematic manner. In addition to genes previously linked to genome instability, our approach implicates novel genes involved in chromatin remodeling and meiosis in affecting the rate of SSA-mediated deletion formation in the presence or absence of stress conditions induced by DNA-damaging agents. We closely examined two candidate genes, the chromatin remodeling gene IOC4 and the meiosis-related gene MSH4, which when knocked-out resulted in gene expression alterations affecting genes involved in cell division and chromosome organization, as well as DNA repair and recombination, respectively. Our high-throughput approach facilitates the systematic identification of processes linked to the formation of a major class of genetic variation.

  5. Managing Single-Stranded DNA during Replication Stress in Fission Yeast

    Directory of Open Access Journals (Sweden)

    Sarah A. Sabatinos

    2015-09-01

    Full Text Available Replication fork stalling generates a variety of responses, most of which cause an increase in single-stranded DNA. ssDNA is a primary signal of replication distress that activates cellular checkpoints. It is also a potential source of genome instability and a substrate for mutation and recombination. Therefore, managing ssDNA levels is crucial to chromosome integrity. Limited ssDNA accumulation occurs in wild-type cells under stress. In contrast, cells lacking the replication checkpoint cannot arrest forks properly and accumulate large amounts of ssDNA. This likely occurs when the replication fork polymerase and helicase units are uncoupled. Some cells with mutations in the replication helicase (mcm-ts mimic checkpoint-deficient cells, and accumulate extensive areas of ssDNA to trigger the G2-checkpoint. Another category of helicase mutant (mcm4-degron causes fork stalling in early S-phase due to immediate loss of helicase function. Intriguingly, cells realize that ssDNA is present, but fail to detect that they accumulate ssDNA, and continue to divide. Thus, the cellular response to replication stalling depends on checkpoint activity and the time that replication stress occurs in S-phase. In this review we describe the signs, signals, and symptoms of replication arrest from an ssDNA perspective. We explore the possible mechanisms for these effects. We also advise the need for caution when detecting and interpreting data related to the accumulation of ssDNA.

  6. Quantitation of ultraviolet-induced single-strand breaks using oligonucleotide chip

    International Nuclear Information System (INIS)

    Pal, Sukdeb; Kim, Min Jung; Choo, Jaebum; Kang, Seong Ho; Lee, Kyeong-Hee; Song, Joon Myong

    2008-01-01

    A simple, accurate and robust methodology was established for the direct quantification of ultraviolet (UV)-induced single-strand break (SSB) using oligonucleotide chip. Oligonucleotide chips were fabricated by covalently anchoring the fluorescent-labeled ssDNAs onto silicon dioxide chip surfaces. Assuming that the possibility of more than one UV-induced SSB to be generated in a small oligonucleotide is extremely low, SSB formation was investigated quantifying the endpoint probe density by fluorescence measurement upon UV irradiation. The SSB yields obtained based on the highly sensitive laser-induced fluorometric determination of fluorophore-labeled oligonucleotides were found to coincide well with that predicted from a theoretical extrapolation of the results obtained for plasmid DNAs using conventional agarose gel electrophoresis. The developed method has the potential to serve as a high throughput, sample-thrifty, and time saving tool to realize more realistic, and direct quantification of radiation and chemical-induced strand breaks. It will be especially useful for determining the frequency of SSBs or lesions convertible to SSBs by specific cleaving reagents or enzymes

  7. Detection of hepatitis A virus by hybridization with single-stranded RNA probes

    International Nuclear Information System (INIS)

    Xi, J.; Estes, M.K.; Metcalf, T.G.

    1987-01-01

    An improved method of dot-blot hybridization to detect hepatitis A virus (HAV) was developed with single-stranded RNA (ssRNA) probes. Radioactive and nonradioactive ssRNA probes were generated by in vitro transcription of HAV templates inserted into the plasmid pGEM-1. 32 P-labeled ssRNA probes were at least eightfold more sensitive than the 32 P-labeled double-stranded cDNA counterparts, whereas biotin-labeled ssRNA probes showed a sensitivity comparable with that of the 32 P-labeled double-stranded cDNA counterparts. Hybridization of HAV with the ssRNA probes at high stringency revealed specific reactions with a high signal-to-noise ratio. The differential hybridization reactions seen with probes of positive and negative sense (compared with HAV genomic RNA) were used to detect HAV in clinical and field samples. A positive/negative ratio was introduced as an indicator that permitted an semiquantitative expression of a positive HAV reaction. Good agreement of this indicator was observed with normal stool samples and with HAV-seeded samples. By using this system, HAV was detected in estuarine and freshwater samples collected from a sewage-polluted bayou in Houston and a saltwater tributary of Galveston Bay

  8. Viral single-strand DNA induces p53-dependent apoptosis in human embryonic stem cells.

    Science.gov (United States)

    Hirsch, Matthew L; Fagan, B Matthew; Dumitru, Raluca; Bower, Jacquelyn J; Yadav, Swati; Porteus, Matthew H; Pevny, Larysa H; Samulski, R Jude

    2011-01-01

    Human embryonic stem cells (hESCs) are primed for rapid apoptosis following mild forms of genotoxic stress. A natural form of such cellular stress occurs in response to recombinant adeno-associated virus (rAAV) single-strand DNA genomes, which exploit the host DNA damage response for replication and genome persistence. Herein, we discovered a unique DNA damage response induced by rAAV transduction specific to pluripotent hESCs. Within hours following rAAV transduction, host DNA damage signaling was elicited as measured by increased gamma-H2AX, ser15-p53 phosphorylation, and subsequent p53-dependent transcriptional activation. Nucleotide incorporation assays demonstrated that rAAV transduced cells accumulated in early S-phase followed by the induction of apoptosis. This lethal signaling sequalae required p53 in a manner independent of transcriptional induction of Puma, Bax and Bcl-2 and was not evident in cells differentiated towards a neural lineage. Consistent with a lethal DNA damage response induced upon rAAV transduction of hESCs, empty AAV protein capsids demonstrated no toxicity. In contrast, DNA microinjections demonstrated that the minimal AAV origin of replication and, in particular, a 40 nucleotide G-rich tetrad repeat sequence, was sufficient for hESC apoptosis. Our data support a model in which rAAV transduction of hESCs induces a p53-dependent lethal response that is elicited by a telomeric sequence within the AAV origin of replication.

  9. Viral single-strand DNA induces p53-dependent apoptosis in human embryonic stem cells.

    Directory of Open Access Journals (Sweden)

    Matthew L Hirsch

    Full Text Available Human embryonic stem cells (hESCs are primed for rapid apoptosis following mild forms of genotoxic stress. A natural form of such cellular stress occurs in response to recombinant adeno-associated virus (rAAV single-strand DNA genomes, which exploit the host DNA damage response for replication and genome persistence. Herein, we discovered a unique DNA damage response induced by rAAV transduction specific to pluripotent hESCs. Within hours following rAAV transduction, host DNA damage signaling was elicited as measured by increased gamma-H2AX, ser15-p53 phosphorylation, and subsequent p53-dependent transcriptional activation. Nucleotide incorporation assays demonstrated that rAAV transduced cells accumulated in early S-phase followed by the induction of apoptosis. This lethal signaling sequalae required p53 in a manner independent of transcriptional induction of Puma, Bax and Bcl-2 and was not evident in cells differentiated towards a neural lineage. Consistent with a lethal DNA damage response induced upon rAAV transduction of hESCs, empty AAV protein capsids demonstrated no toxicity. In contrast, DNA microinjections demonstrated that the minimal AAV origin of replication and, in particular, a 40 nucleotide G-rich tetrad repeat sequence, was sufficient for hESC apoptosis. Our data support a model in which rAAV transduction of hESCs induces a p53-dependent lethal response that is elicited by a telomeric sequence within the AAV origin of replication.

  10. Biophysical characterization of the association of histones with single-stranded DNA.

    Science.gov (United States)

    Wang, Ying; van Merwyk, Luis; Tönsing, Katja; Walhorn, Volker; Anselmetti, Dario; Fernàndez-Busquets, Xavier

    2017-11-01

    Despite the profound current knowledge of the architecture and dynamics of nucleosomes, little is known about the structures generated by the interaction of histones with single-stranded DNA (ssDNA), which is widely present during replication and transcription. Non-denaturing gel electrophoresis, transmission electron microscopy, atomic force microscopy, magnetic tweezers. Histones have a high affinity for ssDNA in 0.15M NaCl ionic strength, with an apparent binding constant similar to that calculated for their association with double-stranded DNA (dsDNA). The length of DNA (number of nucleotides in ssDNA or base pairs in dsDNA) associated with a fixed core histone mass is the same for both ssDNA and dsDNA. Although histone-ssDNA complexes show a high tendency to aggregate, nucleosome-like structures are formed at physiological salt concentrations. Core histones are able to protect ssDNA from digestion by micrococcal nuclease, and a shortening of ssDNA occurs upon its interaction with histones. The purified (+) strand of a cloned DNA fragment of nucleosomal origin has a higher affinity for histones than the purified complementary (-) strand. At physiological ionic strength histones have high affinity for ssDNA, possibly associating with it into nucleosome-like structures. In the cell nucleus histones may spontaneously interact with ssDNA to facilitate their participation in the replication and transcription of chromatin. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. New Method for Differentiation of Granuloviruses (Betabaculoviruses Based on Multitemperature Single Stranded Conformational Polymorphism

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    Martyna Krejmer-Rabalska

    2017-12-01

    Full Text Available Baculoviruses have been used as biopesticides for decades. Recently, due to the excessive use of chemical pesticides there is a need for finding new agents that may be useful in biological protection. Sometimes few isolates or species are discovered in one host. In the past few years, many new baculovirus species have been isolated from environmental samples, thoroughly characterized and thanks to next generation sequencing methods their genomes are being deposited in the GenBank database. Next generation sequencing (NGS methodology is the most certain way of detection, but it has many disadvantages. During our studies, we have developed a method based on Polymerase chain reaction (PCR followed by Multitemperature Single Stranded Conformational Polymorphism (MSSCP which allows for distinguishing new granulovirus isolates in only a few hours and at low-cost. On the basis of phylogenetic analysis of betabaculoviruses, representative species have been chosen. The alignment of highly conserved genes—granulin and late expression factor-9, was performed and the degenerate primers were designed to amplify the most variable, short DNA fragments flanked with the most conserved sequences. Afterwards, products of PCR reaction were analysed by MSSCP technique. In our opinion, the proposed method may be used for screening of new isolates derived from environmental samples.

  12. Delayed repair of DNA single-strand breaks does not increase cytogenetic damage

    International Nuclear Information System (INIS)

    Morgan, W.F.; Djordjevic, M.C.; Jostes, R.F.; Pantelias, G.E.

    1985-01-01

    DNA damage and cytogenetic effects of ionizing radiation were investigated in Chinese hamster ovary (CHO) cells and unstimulated human peripheral blood lymphocytes. DNA damage and repair were analysed by alkaline elution under conditions that predominantly measured DNA single-strand breaks (ssb). X-radiation (2.5 Gy) induced ssb in both CHO cells and unstimulated lymphocytes, and the breaks were repaired within 30 and 90 min, respectively. This rapid repair was delayed by the poly(ADP-ribose) polymerase inhibitor, 3-aminobenzamide (3AB). The cytogenetic effects of the 3AB-induced delay in DNA repair were examined by analysing sister chromatid exchange (SCE) frequency in CHO cells and fragmentation of prematurely condensed chromosomes (PCC) in unstimulated human lymphocytes after 2.5 Gy of X-rays. Although 3AB delayed the rejoining of DNA ssb, this delay did not result in increased cytogenetic damage manifested as either SCE or fragmentation of PCC. These results indicate that the rapidly rejoining DNA ssb are not important in the production of chromosome damage. (author)

  13. Capillary electrophoresis single-strand conformation polymorphism for the monitoring of gastrointestinal microbiota of chicken flocks.

    Science.gov (United States)

    Pissavin, C; Burel, C; Gabriel, I; Beven, V; Mallet, S; Maurice, R; Queguiner, M; Lessire, M; Fravalo, P

    2012-09-01

    The objective of the present study was to evaluate the capillary electrophoresis single-strand conformation polymorphism (CE-SSCP) to characterize poultry gut microbiota and the ability of this molecular method to detect modifications related to rearing conditions to be used as an epidemiological tool. The V3 region of the 16S rRNA gene was selected as the PCR target. Our results showed that this method provides reproducible data. The microbiota analysis of individuals showed that variability between individual fingerprints was higher for ileum and cloaca than for ceca. However, pooling the samples decreased this variability. To estimate the variability within and between farms, we compared molecular gut patterns of animals from the same hatchery reared under similar conditions and fed the same diet in 2 separate farms. Total aerobic bacteria, coliforms, and lactic acid bacteria were enumerated using conventional bacteriological methods. A significant difference was observed for coliforms present in the ceca and the cloaca depending on the farm. Ileal contents fingerprints were more closely related to those of cloacal contents than to those of ceca contents. When comparing samples from the 2 farms, a specific microbiota was highlighted for each farm. For each gut compartment, the microbiota fingerprints were joined in clusters according to the farm. Thus, this rapid and potentially high-throughput method to obtain gut flora fingerprints is sensitive enough to detect a "farm effect" on the balance of poultry gut microbiota despite the birds being fed the same regimens and reared under similar conditions.

  14. QUANTITATION OF INTRACELLULAR NAD(P)H IN LIVING CELLS CAN MONITOR AN IMBALANCE OF DNA SINGLE STRAND BREAK REPAIR IN REAL TIME

    Science.gov (United States)

    Quantitation of intracellular NAD(P)H in living cells can monitor an imbalance of DNA single strand break repair in real time.ABSTRACTDNA single strand breaks (SSBs) are one of the most frequent DNA lesions in genomic DNA generated either by oxidative stress or du...

  15. Synergistic effect of wire bending and salivary pH on surface properties and mechanical properties of orthodontic stainless steel archwires.

    Science.gov (United States)

    Hobbelink, Marieke G; He, Yan; Xu, Jia; Xie, Huixu; Stoll, Richard; Ye, Qingsong

    2015-01-01

    The aim of this study was to investigate the corrosive behaviour of stainless steel archwires in a more clinically relevant way by bending and exposing to various pH. One hundred and twenty pieces of rectangular stainless steel wires (0.43 × 0.64 mm) were randomly assigned into four groups. In each group, there were 15 pieces of bent wires and 15 straight ones. Prior to measurements of the wires, as individual experimental groups (group 1, 2, and 3), the wires were exposed to artificial saliva for 4 weeks at pH 5.6, 6.6, and 7.6, respectively. A control group of wires (group 4) remained in air for the same period of time before sent for measurements. Surface roughness (Ra-value) was measured by a profilometer. Young's modulus and maximum force were determined by a four-point flexural test apparatus. Scanning electron microscopy was used to observe the surface morphology of straight wire. Differences between groups were examined using a two-way analysis of variance (ANOVA). Mean surface roughness values, flexural Young's moduli, and maximum force values of bent wires are significantly different from those of the straight wires, which was the main effect of wire bending, ignoring the influence of pH. A significant effect was found between Ra-values regarding the main effect of pH, ignoring the influence of shape. There was a significant interaction effect of bending and pH on flexural Young's moduli of stainless steel archwires, while pH did not show much impact on the maximum force values of those stainless steel wires. Bigger surface irregularities were seen on SEM images of straight wires immersed in artificial saliva at pH 5.6 compared to artificial saliva at other pH values. Surface depth (Rz) was more sensitive than Ra in revealing surface roughness, both measured from 3D reconstructed SEM images. Ra showed a comparable result of surface roughness to Ra-value measured by the profilometer. Bending has a significant influence on surface roughness and mechanical

  16. Selection and Characterization of Single Stranded DNA Aptamers for the Hormone Abscisic Acid

    Science.gov (United States)

    Gonzalez, Victor M.; Millo, Enrico; Sturla, Laura; Vigliarolo, Tiziana; Bagnasco, Luca; Guida, Lucrezia; D'Arrigo, Cristina; De Flora, Antonio; Salis, Annalisa; Martin, Elena M.; Bellotti, Marta; Zocchi, Elena

    2013-01-01

    The hormone abscisic acid (ABA) is a small molecule involved in pivotal physiological functions in higher plants. Recently, ABA has been also identified as an endogenous hormone in mammals, regulating different cell functions including inflammatory processes, stem cell expansion, insulin release, and glucose uptake. Aptamers are short, single-stranded (ss) oligonucleotidesable to recognize target molecules with high affinity. The small size of the ABA molecule represented a challenge for aptamer development and the aim of this study was to develop specific anti-ABA DNA aptamers. Biotinylated abscisic acid (bio-ABA) was immobilized on streptavidin-coated magnetic beads. DNA aptamers against bio-ABA were selected with 7 iterative rounds of the systematic evolution of ligands by exponential enrichment method (SELEX), each round comprising incubation of the ABA-binding beads with the ssDNA sequences, DNA elution, electrophoresis, and polymerase chain reaction (PCR) amplification. The PCR product was cloned and sequenced. The binding affinity of several clones was determined using bio-ABA immobilized on streptavidin-coated plates. Aptamer 2 and aptamer 9 showed the highest binding affinity, with dissociation constants values of 0.98±0.14 μM and 0.80±0.07 μM, respectively. Aptamers 2 and 9 were also able to bind free, unmodified ABA and to discriminate between different ABA enantiomers and isomers. Our findings indicate that ssDNA aptamers can selectively bind ABA and could be used for the development of ABA quantitation assays. PMID:23971905

  17. Multicopy Single-Stranded DNA Directs Intestinal Colonization of Enteric Pathogens

    Energy Technology Data Exchange (ETDEWEB)

    Elfenbein, Johanna R.; Knodler, Leigh A.; Nakayasu, Ernesto S.; Ansong, Charles; Brewer, Heather M.; Bogomolnaya, Lydia; Adams, L. Garry; McClelland, Michael; Adkins, Joshua N.; Andrews-Polymenis, Helene L.; Fang, Ferric C.

    2015-09-14

    Multicopy single-stranded DNAs (msDNAs) are hybrid RNA-DNA molecules encoded on retroelements called retrons and produced by the action of retron reverse transcriptases. Retrons are widespread in bacteria but the natural function of msDNA has remained elusive despite 30 years of study. The major roadblock to elucidation of the function of these unique molecules has been the lack of any identifiable phenotypes for mutants unable to make msDNA. We report that msDNA of the zoonotic pathogen Salmonella Typhimurium is necessary for colonization of the intestine. Similarly, we observed a defect in intestinal persistence in an enteropathogenic E. coli mutant lacking its retron reverse transcriptase. Under anaerobic conditions in the absence of msDNA, proteins of central anaerobic metabolism needed for Salmonella colonization of the intestine are dysregulated. We show that the msDNA-deficient mutant can utilize nitrate but not other alternate electron acceptors in anaerobic conditions. Consistent with the availability of nitrate in the inflamed gut, a neutrophilic inflammatory response partially rescued the ability of a mutant lacking msDNA to colonize the intestine. These findings together indicate that the mechanistic basis of msDNA function during Salmonella colonization of the intestine is proper production of proteins needed for anaerobic metabolism. We further conclude that a natural function of msDNA is to regulate protein abundance, the first attributable function for any msDNA. Our data provide novel insight into the function of this mysterious molecule that likely represents a new class of regulatory molecules.

  18. Screening for mutations in human alpha-globin genes by nonradioactive single-strand conformation polymorphism

    Directory of Open Access Journals (Sweden)

    Jorge S.B.

    2003-01-01

    Full Text Available Point mutations and small insertions or deletions in the human alpha-globin genes may produce alpha-chain structural variants and alpha-thalassemia. Mutations can be detected either by direct DNA sequencing or by screening methods, which select the mutated exon for sequencing. Although small (about 1 kb, 3 exons and 2 introns, the alpha-globin genes are duplicate (alpha2 and alpha1 and highy G-C rich, which makes them difficult to denature, reducing sequencing efficiency and causing frequent artifacts. We modified some conditions for PCR and electrophoresis in order to detect mutations in these genes employing nonradioactive single-strand conformation polymorphism (SSCP. Primers previously described by other authors for radioactive SSCP and phast-SSCP plus denaturing gradient gel electrophoresis were here combined and the resultant fragments (6 new besides 6 original per alpha-gene submitted to silver staining SSCP. Nine structural and one thalassemic mutations were tested, under different conditions including two electrophoretic apparatus (PhastSystem(TM and GenePhor(TM, Amersham Biosciences, different polyacrylamide gel concentrations, run temperatures and denaturing agents, and entire and restriction enzyme cut fragments. One hundred percent of sensitivity was achieved with four of the new fragments formed, using the PhastSystem(TM and 20% gels at 15ºC, without the need of restriction enzymes. This nonradioactive PCR-SSCP approach showed to be simple, rapid and sensitive, reducing the costs involved in frequent sequencing repetitions and increasing the reliability of the results. It can be especially useful for laboratories which do not have an automated sequencer.

  19. A high throughput system for the preparation of single stranded templates grown in microculture.

    Science.gov (United States)

    Kolner, D E; Guilfoyle, R A; Smith, L M

    1994-01-01

    A high throughput system for the preparation of single stranded M13 sequencing templates is described. Supernatants from clones grown in 48-well plates are treated with a chaotropic agent to dissociate the phage coat protein. Using a semi-automated cell harvester, the free nucleic acid is bound to a glass fiber filter in the presence of chaotrope and then washed with ethanol by aspiration. Individual glass fiber discs are punched out on the cell harvester and dried briefly. The DNA samples are then eluted in water by centrifugation. The processing time from 96 microcultures to sequence quality templates is approximately 1 hr. Assuming the ability to sequence 400 bases per clone, a 0.5 megabase per day genome sequencing facility will require 6250 purified templates a week. Toward accomplishing this goal we have developed a procedure which is a modification of a method that uses a chaotropic agent and glass fiber filter (Kristensen et al., 1987). By exploiting the ability of a cell harvester to uniformly aspirate and wash 96 samples, a rapid system for high quality template preparation has been developed. Other semi-automated systems for template preparation have been developed using commercially available robotic workstations like the Biomek (Mardis and Roe, 1989). Although minimal human intervention is required, processing time is at least twice as long. Custom systems based on paramagnetic beads (Hawkins et al., 1992) produce DNA in insufficient quantity for direct sequencing and therefore require cycle sequencing. These systems require custom programing, have a fairly high initial cost and have not proven to be as fast as the method reported here.

  20. Identification of five novel FBN1 mutations by non-radioactive single-strand conformation analysis

    Energy Technology Data Exchange (ETDEWEB)

    Liu, W.; Qian, C.; Comeau, K.; Francke, U. [Stanford Univ. Medical Center, Stanford, CA (United States)

    1994-09-01

    Marfan syndrome (MFS), one of the most common genetic disorders of connective tissue, is characterized by variable manifestations in skeletal, cardiovascular and ocular systems. Mutations in the fibrillin gene on chromosome 15 (FBN1) have been shown to cause MFS. To examine the relationship between FBN1 gene mutations, fibrillin protein function and MFS phenotypes, we screened for alternations in the fibrillin coding sequence in fibroblast derived cDNA from MFS patients. To date, abnormally migrating bands in more than 20 unrelated MFS patients have been identified by using non-radioactive single-strand conformation analysis and silver staining. Five altered bands have been directly sequenced. Two missense mutations and three splice site mutations have been identified. Both missense mutations substitute another amino acid for a cysteine residue (C1402W and C1672R) in EGF-like motifs of the fibrillin polypeptide chain. The two splice site mutations are at nucleotide positions 6994+1 (G{yields}A), and 7205-2 (A{yields}G) and result in in-frame skipping of exon 56 and 58, respectively. Skipping of exon 56 occurs in 50% of mutant transcripts. Use of a cryptic splice site 51 bp upstream of the normal donor site results in half of the mutant transcripts containing part of exon 56. Both products contain in-frame deletions. Another splice site mutation, identified by exon screening from patient genomic DNA using intron primers, is at nucleotide position 2293+2 (T{yields}A), but the predicted exon skipping has not been detected at the RT-PCR level. This may be due to instability of the mutant transcript. Including the mutations reported here, a total of 8 out of 36 published FBN1 gene mutations involve exon skipping. It may be inferred that FBN1 exon skipping plays an important pathogenic role in MFS.

  1. Carboplatin enhances the production and persistence of radiation-induced DNA single-strand breaks

    International Nuclear Information System (INIS)

    Yang, L.; Douple, E.B.; O'Hara, J.A.; Wang, H.J.

    1995-01-01

    Fluorometric analysis of DNA unwinding and alkaline elution were used to investigate the production and persistence of DNA single-strand breaks (SSBs) in Chinese hamster V79 and xrs-5 cells treated with the chemotherapeutic agent carboplatin in combination with radiation. Carboplatin was administered to cells before irradiation in hypoxic conditions, or the drug was added immediately after irradiation during the postirradiation recovery period in air. The results of DNA unwinding studies suggest that carboplatin enhances the production of radiation-induced SSBs in hypoxic V79 cells and xrs-5 cells by a factor of 1.86 and 1.83, respectively, when combined with radiation compared to the SSBs produced by irradiation alone. Carboplatin alone did not produce a measureable number of SSBs. Alkaline elution profiles also indicated that the rate of elution of SSBs was higher in cells treated with the carboplatin is present after irradiation and during the postirradiation recovery period, the rejoining of radiation-induced SSBs by a factor of 1.46 in V79 cells with 20 Gy irradiation and by a factor of 2.02 in xrs-5 cells with 20 Gy irradiation. When carboplatin is present after irradiation and during the postirradiation recovery period, the rejoining of radiation-induced SSBs is inhibited during this postirradiation incubation period (radiopotentiation) with a relative inhibition factor at 1 h postirradiation of 1.25 in V79 cells and 1.15 in xrs-5 cells. An increased production and persistence of SSBs resulting from the interaction of carboplatin with radiation may be an important step in the mechanism responsible for the potentiated cell killing previously from studies in animal tumors and in cultured cells. 31 refs., 7 figs

  2. Chemo-mechanical pushing of proteins along single-stranded DNA.

    Science.gov (United States)

    Sokoloski, Joshua E; Kozlov, Alexander G; Galletto, Roberto; Lohman, Timothy M

    2016-05-31

    Single-stranded (ss)DNA binding (SSB) proteins bind with high affinity to ssDNA generated during DNA replication, recombination, and repair; however, these SSBs must eventually be displaced from or reorganized along the ssDNA. One potential mechanism for reorganization is for an ssDNA translocase (ATP-dependent motor) to push the SSB along ssDNA. Here we use single molecule total internal reflection fluorescence microscopy to detect such pushing events. When Cy5-labeled Escherichia coli (Ec) SSB is bound to surface-immobilized 3'-Cy3-labeled ssDNA, a fluctuating FRET signal is observed, consistent with random diffusion of SSB along the ssDNA. Addition of Saccharomyces cerevisiae Pif1, a 5' to 3' ssDNA translocase, results in the appearance of isolated, irregularly spaced saw-tooth FRET spikes only in the presence of ATP. These FRET spikes result from translocase-induced directional (5' to 3') pushing of the SSB toward the 3' ssDNA end, followed by displacement of the SSB from the DNA end. Similar ATP-dependent pushing events, but in the opposite (3' to 5') direction, are observed with EcRep and EcUvrD (both 3' to 5' ssDNA translocases). Simulations indicate that these events reflect active pushing by the translocase. The ability of translocases to chemo-mechanically push heterologous SSB proteins along ssDNA provides a potential mechanism for reorganization and clearance of tightly bound SSBs from ssDNA.

  3. Ammonia disinfection of hatchery waste for elimination of single-stranded RNA viruses.

    Science.gov (United States)

    Emmoth, Eva; Ottoson, Jakob; Albihn, Ann; Belák, Sándor; Vinnerås, Björn

    2011-06-01

    Hatchery waste, an animal by-product of the poultry industry, needs sanitation treatment before further use as fertilizer or as a substrate in biogas or composting plants, owing to the potential presence of opportunistic pathogens, including zoonotic viruses. Effective sanitation is also important in viral epizootic outbreaks and as a routine, ensuring high hygiene standards on farms. This study examined the use of ammonia at different concentrations and temperatures to disinfect hatchery waste. Inactivation kinetics of high-pathogenic avian influenza virus H7N1 and low-pathogenic avian influenza virus H5N3, as representatives of notifiable avian viral diseases, were determined in spiked hatchery waste. Bovine parainfluenza virus type 3, feline coronavirus, and feline calicivirus were used as models for other important avian pathogens, such as Newcastle disease virus, infectious bronchitis virus, and avian hepatitis E virus. Bacteriophage MS2 was also monitored as a stable indicator. Coronavirus was the most sensitive virus, with decimal reduction (D) values of 1.2 and 0.63 h after addition of 0.5% (wt/wt) ammonia at 14 and 25°C, respectively. Under similar conditions, high-pathogenic avian influenza H7N1 was the most resistant, with D values of 3.0 and 1.4 h. MS2 was more resistant than the viruses to all treatments and proved to be a suitable indicator of viral inactivation. The results indicate that ammonia treatment of hatchery waste is efficient in inactivating enveloped and naked single-stranded RNA viruses. Based on the D values and confidence intervals obtained, guidelines for treatment were proposed, and one was successfully validated at full scale at a hatchery, with MS2 added to hatchery waste.

  4. Stabilization of Pt nanoparticles by single stranded DNA and the binary assembly of Au and Pt nanoparticles without hybridization

    International Nuclear Information System (INIS)

    Yang, J.; Lee, Jim Yang; Too, Heng-Phon; Chow, Gan-Moog; Gan, Leong M.

    2006-01-01

    The non-specific interaction between single stranded DNA (ssDNA) and 12 nm Pt nanoparticles is investigated in this work. The data show a strong and non-specific interaction between the two which can be exploited for the stabilization of Pt nanoparticles in aqueous solutions. Based on the experimental findings, a non-hybridization based protocol to assemble 17 nm Au and Pt nanoparticles (12 nm cubic and 3.6 nm spherical) by single-stranded DNA was developed. Transmission electron microscopy (TEM) and UV-visible spectroscopy confirmed that Au and Pt nanoparticles could be assembled by the non-specific interaction in an orderly manner. The experimental results also caution against the potential pitfalls in using DNA melting point analysis to infer metal nanoparticle assembly by DNA hybridization

  5. Orthodontics at a Pivotal Point of Transformation

    Science.gov (United States)

    Mao, Jeremy J.

    2014-01-01

    The profession of orthodontics is projected to face a multitude of challenges. Do cyclic forces accelerate the rate of tooth movement and hence the speed of orthodontic treatment? Would bioengineered cementum and dentine be a solution to root resorption? What would orthodontics be like when bioengineered periodontal ligament and alveolar bone become clinical practice, or one day, entire teeth are bioengineered? Would it be possible to selectively differentiate stem cells into osteoblasts or osteoclasts by either static or cyclic forces? What is the new demand on orthodontic expertise with increasingly automated appliances? What will be the impact of the next generation of dental implants or rapid prototyped crowns on orthodontics? A century ago, Edward Angle’s practice of fixed appliances, along with other seminal contributions, such as functional appliances, established the profession of orthodontics. Today, the biophysical principles of orthodontics remain largely unchanged from Angle’s era, despite incremental refinements of brackets and wires. The paucity of fundamental innovations in orthodontics for decades presents intrinsic risks for the profession. This review will identify challenges for contemporary orthodontics and delineate strategies for the profession to evolve in an era of unprecedented scientific and technological advances, and serve as a call to action for the orthodontic profession. PMID:25018618

  6. Explanation for excessive DNA single-strand breaks and endogenous repair foci in pluripotent mouse embryonic stem cells.

    Science.gov (United States)

    Banáth, J P; Bañuelos, C A; Klokov, D; MacPhail, S M; Lansdorp, P M; Olive, P L

    2009-05-01

    Pluripotent mouse embryonic stem cells (mES cells) exhibit approximately 100 large gammaH2AX repair foci in the absence of measurable numbers of DNA double-strand breaks. Many of these cells also show excessive numbers of DNA single-strand breaks (>10,000 per cell) when analyzed using the alkaline comet assay. To understand the reasons for these unexpected observations, various methods for detecting DNA strand breaks were applied to wild-type mES cells and to mES cells lacking H2AX, ATM, or DNA-PKcs. H2AX phosphorylation and expression of other repair complexes were measured using flow and image analysis of antibody-stained cells. Results indicate that high numbers of endogenous gammaH2AX foci and single-strand breaks in pluripotent mES cells do not require ATM or DNA-PK kinase activity and appear to be associated with global chromatin decondensation rather than pre-existing DNA damage. This will limit applications of gammaH2AX foci analysis in mES cells to relatively high levels of initial or residual DNA damage. Excessive numbers of single-strand breaks in the alkaline comet assay can be explained by the vulnerability of replicating chromatin in mES cells to osmotic shock. This suggests that caution is needed in interpreting results with the alkaline comet assay when applied to certain cell types or after treatment with agents that make chromatin vulnerable to osmotic changes. Differentiation of mES cells caused a reduction in histone acetylation, gammaH2AX foci intensity, and DNA single-strand breakage, providing a link between chromatin structural organization, excessive gammaH2AX foci, and sensitivity of replicating mES cell chromatin to osmotic shock.

  7. Near-Complete Genome Sequence of a Novel Single-Stranded RNA Virus Discovered in Indoor Air.

    Science.gov (United States)

    Rosario, Karyna; Fierer, Noah; Breitbart, Mya

    2018-03-22

    Viral metagenomic analysis of heating, ventilation, and air conditioning (HVAC) filters recovered the near-complete genome sequence of a novel virus, named HVAC-associated R NA v irus 1 (HVAC-RV1). The HVAC-RV1 genome is most similar to those of picorna-like viruses identified in arthropods but encodes a small domain observed only in negative-sense single-stranded RNA viruses. Copyright © 2018 Rosario et al.

  8. Alterations in the nuclear matrix protein mass correlate with heat-induced inhibition of DNA single-strand-break repair

    International Nuclear Information System (INIS)

    Warters, R.L.; Brizgys, L.M.; Lyons, B.W.

    1987-01-01

    The total protein mass co-isolating with the nuclear matrix or nucleoid from Chinese hamster ovary (CHO) cells was observed to increase in heated cells as a function of increasing exposure temperature between 43 0 C and 45 0 C or of exposure time at any temperature. The sedimentation distance of the CHO cell nucleoid in sucrose gradients increased with increasing exposure time at 45 0 C. Both these nuclear alterations correlated in a log-linear manner with heat-induced inhibition of DNA strand break repair. A two-fold threshold increase in nuclear matrix protein mass preceded any substantial inhibition of repair of DNA single-strand breaks. When preheated cells were incubated at 37 0 C the nuclear matrix protein mass and nucleoid sedimentation recovered with a half-time of about 5 h, while DNA single-strand-break repair recovered with a half-time of about 2 h. When preheated cells were placed at 41 0 C a further increase was observed in the nuclear matrix protein mass and the half-time of DNA strand break repair, while nucleoid sedimentation recovered toward control values. These results implicate alterations in the protein mass of the nuclear matrix in heat-induced inhibition of repair of DNA single-strand breaks. (author)

  9. Radiation-induced DNA single-strand scission and its rejoining in spermatogonia and spermatozoa of mouse

    International Nuclear Information System (INIS)

    Ono, T.; Okada, S.

    1977-01-01

    Gamma-ray-induced DNA single-strand scissions and the ability to repair the scissions in spermatogonia from young mice and in spermatozoa from adult mice were studied quantitatively by an alkaline sucrose density-gradient centrifugation method. The average size of DNAs in non-irradiated spermatogonia was 2.6-3.0xx10 8 daltons, similar to those of a spermatid-rich population, and the size of DNA in non-irradiated spermatozoa was 1.2x10 8 daltons. In spermatogonia, the radiosensitivity of DNA was 0.42 single-strand breaks/10 12 daltons of DNA/rad in oxic conditions and only 0.24 under anoxic conditions. In spermatozoa the break efficiency of DNA was 0.22 single-strand breaks/10 12 daltons of DNA/rad under oxic conditions and altered little under anoxic irradiation. The DNA scissions were efficiently repaired in spermatogonia within 10 min, whereas the breaks in spermatozoa were not rejoined at all even after two days of post-irradiation time. The radiosensitivities of DNA, repair capability and non- and/or slowreparable DNA scissions were compared in spermatogonium-rich, spermatid-rich and spermatozoanrich populations

  10. Role of electrostatics in the assembly pathway of a single-stranded RNA virus.

    Science.gov (United States)

    Garmann, Rees F; Comas-Garcia, Mauricio; Koay, Melissa S T; Cornelissen, Jeroen J L M; Knobler, Charles M; Gelbart, William M

    2014-09-01

    We have recently discovered (R. D. Cadena-Nava et al., J. Virol. 86:3318-3326, 2012, doi:10.1128/JVI.06566-11) that the in vitro packaging of RNA by the capsid protein (CP) of cowpea chlorotic mottle virus is optimal when there is a significant excess of CP, specifically that complete packaging of all of the RNA in solution requires sufficient CP to provide charge matching of the N-terminal positively charged arginine-rich motifs (ARMS) of the CPs with the negatively charged phosphate backbone of the RNA. We show here that packaging results from the initial formation of a charge-matched protocapsid consisting of RNA decorated by a disordered arrangement of CPs. This protocapsid reorganizes into the final, icosahedrally symmetric nucleocapsid by displacing the excess CPs from the RNA to the exterior surface of the emerging capsid through electrostatic attraction between the ARMs of the excess CP and the negative charge density of the capsid exterior. As a test of this scenario, we prepare CP mutants with extra and missing (relative to the wild type) cationic residues and show that a correspondingly smaller and larger excess, respectively, of CP is needed for complete packaging of RNA. Cowpea chlorotic mottle virus (CCMV) has long been studied as a model system for the assembly of single-stranded RNA viruses. While much is known about the electrostatic interactions within the CCMV virion, relatively little is known about these interactions during assembly, i.e., within intermediate states preceding the final nucleocapsid structure. Theoretical models and coarse-grained molecular dynamics simulations suggest that viruses like CCMV assemble by the bulk adsorption of CPs onto the RNA driven by electrostatic attraction, followed by structural reorganization into the final capsid. Such a mechanism facilitates assembly by condensing the RNA for packaging while simultaneously concentrating the local density of CP for capsid nucleation. We provide experimental evidence of

  11. Functional characterization of an alkaline exonuclease and single strand annealing protein from the SXT genetic element of Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Huang Jian-dong

    2011-04-01

    Full Text Available Abstract Background SXT is an integrating conjugative element (ICE originally isolated from Vibrio cholerae, the bacterial pathogen that causes cholera. It houses multiple antibiotic and heavy metal resistance genes on its ca. 100 kb circular double stranded DNA (dsDNA genome, and functions as an effective vehicle for the horizontal transfer of resistance genes within susceptible bacterial populations. Here, we characterize the activities of an alkaline exonuclease (S066, SXT-Exo and single strand annealing protein (S065, SXT-Bet encoded on the SXT genetic element, which share significant sequence homology with Exo and Bet from bacteriophage lambda, respectively. Results SXT-Exo has the ability to degrade both linear dsDNA and single stranded DNA (ssDNA molecules, but has no detectable endonuclease or nicking activities. Adopting a stable trimeric arrangement in solution, the exonuclease activities of SXT-Exo are optimal at pH 8.2 and essentially require Mn2+ or Mg2+ ions. Similar to lambda-Exo, SXT-Exo hydrolyzes dsDNA with 5'- to 3'-polarity in a highly processive manner, and digests DNA substrates with 5'-phosphorylated termini significantly more effectively than those lacking 5'-phosphate groups. Notably, the dsDNA exonuclease activities of both SXT-Exo and lambda-Exo are stimulated by the addition of lambda-Bet, SXT-Bet or a single strand DNA binding protein encoded on the SXT genetic element (S064, SXT-Ssb. When co-expressed in E. coli cells, SXT-Bet and SXT-Exo mediate homologous recombination between a PCR-generated dsDNA fragment and the chromosome, analogous to RecET and lambda-Bet/Exo. Conclusions The activities of the SXT-Exo protein are consistent with it having the ability to resect the ends of linearized dsDNA molecules, forming partially ssDNA substrates for the partnering SXT-Bet single strand annealing protein. As such, SXT-Exo and SXT-Bet may function together to repair or process SXT genetic elements within infected V

  12. Modifications in straight wire treatment.

    Science.gov (United States)

    Cardona, Alvin

    2010-01-01

    Orthodontic treatments have been modified with each new generation of clinicians. Today the emphasis is on facial esthetics and healthy temporomandibular joints. With orthopedic treatment, we can develop dental arches to get the necessary space to align the teeth and we can reach adequate function and esthetics, all within relatively good stability. By combining two-phase treatment with low friction fixed orthodontics and super elastic wires we produce light but continuous forces and we can provide better treatment than before. These types of forces cause physiological and functional orthopedic orthodontic reactions. The purpose of this article is to demonstrate our fixed orthopedic and orthodontic approach called "Modified Straight Wire" or "Physiologic Arch Technique." This technique is very successful with our patients because it can exert slow and continuous forces with minimal patient cooperation.

  13. Orthodontic scars

    Directory of Open Access Journals (Sweden)

    Vinay Reddy

    2012-01-01

    Full Text Available Orthodontic therapy apart from its benefits also has potential risks and limitations in terms of tissue damage. Fortunately, in orthodontics, risks are minimal and infrequent However, all potential risks and limitations should be considered and addressed when making the decision to undergo orthodontic treatment. Orthodontic treatment carries with it the risk of various types of soft and hard tissue damages (e.g. decalcification of enamel, lacerations, ulcerations, temporomandibular joint disorders, etc-, apart from treatment failure in itself. If correcting a malocclusion is to be of benefit, the advantages offered should outweigh any possible damage- All preventive procedures should be considered during and after orthodontic treatment to restore the normal health of soft and hard tissues. Hence, the orthodontist should be vigilant and prudent enough in assessing and monitoring every aspect of these tissues at any given stage and time in order to achieve a healthy and successful final result.

  14. Identification of a Single Strand Origin of Replication in the Integrative and Conjugative Element ICEBs1 of Bacillus subtilis.

    Directory of Open Access Journals (Sweden)

    Laurel D Wright

    2015-10-01

    Full Text Available We identified a functional single strand origin of replication (sso in the integrative and conjugative element ICEBs1 of Bacillus subtilis. Integrative and conjugative elements (ICEs, also known as conjugative transposons are DNA elements typically found integrated into a bacterial chromosome where they are transmitted to daughter cells by chromosomal replication and cell division. Under certain conditions, ICEs become activated and excise from the host chromosome and can transfer to neighboring cells via the element-encoded conjugation machinery. Activated ICEBs1 undergoes autonomous rolling circle replication that is needed for the maintenance of the excised element in growing and dividing cells. Rolling circle replication, used by many plasmids and phages, generates single-stranded DNA (ssDNA. In many cases, the presence of an sso enhances the conversion of the ssDNA to double-stranded DNA (dsDNA by enabling priming of synthesis of the second DNA strand. We initially identified sso1 in ICEBs1 based on sequence similarity to the sso of an RCR plasmid. Several functional assays confirmed Sso activity. Genetic analyses indicated that ICEBs1 uses sso1 and at least one other region for second strand DNA synthesis. We found that Sso activity was important for two key aspects of the ICEBs1 lifecycle: 1 maintenance of the plasmid form of ICEBs1 in cells after excision from the chromosome, and 2 stable acquisition of ICEBs1 following transfer to a new host. We identified sequences similar to known plasmid sso's in several other ICEs. Together, our results indicate that many other ICEs contain at least one single strand origin of replication, that these ICEs likely undergo autonomous replication, and that replication contributes to the stability and spread of these elements.

  15. Dissociation of single-strand DNA: single-walled carbon nanotube hybrids by Watson-Crick base-pairing.

    Science.gov (United States)

    Jung, Seungwon; Cha, Misun; Park, Jiyong; Jeong, Namjo; Kim, Gunn; Park, Changwon; Ihm, Jisoon; Lee, Junghoon

    2010-08-18

    It has been known that single-strand DNA wraps around a single-walled carbon nanotube (SWNT) by pi-stacking. In this paper it is demonstrated that such DNA is dissociated from the SWNT by Watson-Crick base-pairing with a complementary sequence. Measurement of field effect transistor characteristics indicates a shift of the electrical properties as a result of this "unwrapping" event. We further confirm the suggested process through Raman spectroscopy and gel electrophoresis. Experimental results are verified in view of atomistic mechanisms with molecular dynamics simulations and binding energy analyses.

  16. Packaging signals in single-stranded RNA viruses: nature?s alternative to a purely electrostatic assembly mechanism

    OpenAIRE

    Stockley, Peter G.; Twarock, Reidun; Bakker, Saskia E.; Barker, Amy M.; Borodavka, Alexander; Dykeman, Eric; Ford, Robert J.; Pearson, Arwen R.; Phillips, Simon E. V.; Ranson, Neil A.; Tuma, Roman

    2013-01-01

    The formation of a protective protein container is an essential step in the life-cycle of most viruses. In the case of single-stranded (ss)RNA viruses, this step occurs in parallel with genome packaging in a co-assembly process. Previously, it had been thought that this process can be explained entirely by electrostatics. Inspired by recent single-molecule fluorescence experiments that recapitulate the RNA packaging specificity seen in vivo for two model viruses, we present an alternative the...

  17. Molecular Genetic Characterization of Mutagenesis Using a Highly Sensitive Single-Stranded DNA Reporter System in Budding Yeast.

    Science.gov (United States)

    Chan, Kin

    2018-01-01

    Mutations are permanent alterations to the coding content of DNA. They are starting material for the Darwinian evolution of species by natural selection, which has yielded an amazing diversity of life on Earth. Mutations can also be the fundamental basis of serious human maladies, most notably cancers. In this chapter, I describe a highly sensitive reporter system for the molecular genetic analysis of mutagenesis, featuring controlled generation of long stretches of single-stranded DNA in budding yeast cells. This system is ~100- to ~1000-fold more susceptible to mutation than conventional double-stranded DNA reporters, and is well suited for generating large mutational datasets to investigate the properties of mutagens.

  18. Automated methods for single-stranded DNA isolation and dideoxynucleotide DNA sequencing reactions on a robotic workstation

    International Nuclear Information System (INIS)

    Mardis, E.R.; Roe, B.A.

    1989-01-01

    Automated procedures have been developed for both the simultaneous isolation of 96 single-stranded M13 chimeric template DNAs in less than two hours, and for simultaneously pipetting 24 dideoxynucleotide sequencing reactions on a commercially available laboratory workstation. The DNA sequencing results obtained by either radiolabeled or fluorescent methods are consistent with the premise that automation of these portions of DNA sequencing projects will improve the reproducibility of the DNA isolation and the procedures for these normally labor-intensive steps provides an approach for rapid acquisition of large amounts of high quality, reproducible DNA sequence data

  19. Electroporation and microinjection successfully deliver single-stranded and duplex DNA into live cells as detected by FRET measurements.

    Directory of Open Access Journals (Sweden)

    Rosemary A Bamford

    Full Text Available Förster resonance energy transfer (FRET technology relies on the close proximity of two compatible fluorophores for energy transfer. Tagged (Cy3 and Cy5 complementary DNA strands forming a stable duplex and a doubly-tagged single strand were shown to demonstrate FRET outside of a cellular environment. FRET was also observed after transfecting these DNA strands into fixed and live cells using methods such as microinjection and electroporation, but not when using lipid based transfection reagents, unless in the presence of the endosomal acidification inhibitor bafilomycin. Avoiding the endocytosis pathway is essential for efficient delivery of intact DNA probes into cells.

  20. Versatile Auxiliary Orthodontic Spring for Orthodontic Correction of Impacted Teeth

    Directory of Open Access Journals (Sweden)

    Pavankumar Janardan Vibhute

    2011-01-01

    Full Text Available Malocclusion such as impacted tooth is not uncommon. Many approaches with various auxiliary springs have been reported in literature till date for correction of such malocclusions. They had biomechanical, retentive and stability drawbacks inherent in their designs. This article presents the innovative approach for orthodontic correction of impacted tooth, especially with light force appliance, i.e. Begg′s appliance, where round wires in round molar tubes are used throughout treatment. A versatile auxiliary orthodontic spring (VAOS is fabricated in the 0.018 inch Australian stainless steel round wire, which may be anchored on round molar buccal tube, and desirable force vector may be applied in any of the three dimensions. Fabrication and its clinical application are discussed.

  1. A Novel Method of Coating Orthodontic Archwires with Nanoparticles

    Science.gov (United States)

    Syed, Shibli S; Kulkarni, Dinraj; Todkar, Rohit; Bagul, Ravikiran S; Parekh, Kreena; Bhujbal, Nikita

    2015-01-01

    Background: The major hazard to the orthodontic tooth movement is the friction developing at the bracket wire interface. In the past, there have been various attempts to reduce this friction. We believe that coating the commercially available orthodontic wires with nanoparticles can result in a successful reduction of this friction. The objective of this study is to develop a novel method of coating orthodontic archwires with nanoparticles. Materials and Methods: Stainless steel (Ormco, CA, USA), titanium molybdenum alloy (Ormco, CA, USA) and nickel-titanium (G and H Wire Company, USA) orthodontic wires with a rectangular cross-section dimension of 0.019”× 0.025”, were selected. The wires were later coated with a uniform and smooth nanoparticle film using 100 ml nanocremics. The coating procedure described in this article is a sol-gel thin film dip coating method. Results: The coating procedure was verified by comparing the surface topography of nanocoated archwires with the commercially available archwires in an environmental scanning electron microscope (ESEM). The ESEM images prove that the surface topography of the coated wires was found to be smoother with less surface deteriorations as compared to the commercially available wires. Conclusion: Commercially available orthodontic wires can be successfully coated using a novel method of sol-gel thin film dip coating method. PMID:26028899

  2. Interactive Roles of DNA Helicases and Translocases with the Single-Stranded DNA Binding Protein RPA in Nucleic Acid Metabolism.

    Science.gov (United States)

    Awate, Sanket; Brosh, Robert M

    2017-06-08

    Helicases and translocases use the energy of nucleoside triphosphate binding and hydrolysis to unwind/resolve structured nucleic acids or move along a single-stranded or double-stranded polynucleotide chain, respectively. These molecular motors facilitate a variety of transactions including replication, DNA repair, recombination, and transcription. A key partner of eukaryotic DNA helicases/translocases is the single-stranded DNA binding protein Replication Protein A (RPA). Biochemical, genetic, and cell biological assays have demonstrated that RPA interacts with these human molecular motors physically and functionally, and their association is enriched in cells undergoing replication stress. The roles of DNA helicases/translocases are orchestrated with RPA in pathways of nucleic acid metabolism. RPA stimulates helicase-catalyzed DNA unwinding, enlists translocases to sites of action, and modulates their activities in DNA repair, fork remodeling, checkpoint activation, and telomere maintenance. The dynamic interplay between DNA helicases/translocases and RPA is just beginning to be understood at the molecular and cellular levels, and there is still much to be learned, which may inform potential therapeutic strategies.

  3. Single-stranded γPNAs for in vivo site-specific genome editing via Watson-Crick recognition.

    Science.gov (United States)

    Bahal, Raman; Quijano, Elias; McNeer, Nicole A; Liu, Yanfeng; Bhunia, Dinesh C; Lopez-Giraldez, Francesco; Fields, Rachel J; Saltzman, William M; Ly, Danith H; Glazer, Peter M

    2014-01-01

    Triplex-forming peptide nucleic acids (PNAs) facilitate gene editing by stimulating recombination of donor DNAs within genomic DNA via site-specific formation of altered helical structures that further stimulate DNA repair. However, PNAs designed for triplex formation are sequence restricted to homopurine sites. Herein we describe a novel strategy where next generation single-stranded gamma PNAs (γPNAs) containing miniPEG substitutions at the gamma position can target genomic DNA in mouse bone marrow at mixed-sequence sites to induce targeted gene editing. In addition to enhanced binding, γPNAs confer increased solubility and improved formulation into poly(lactic-co-glycolic acid) (PLGA) nanoparticles for efficient intracellular delivery. Single-stranded γPNAs induce targeted gene editing at frequencies of 0.8% in mouse bone marrow cells treated ex vivo and 0.1% in vivo via IV injection, without detectable toxicity. These results suggest that γPNAs may provide a new tool for induced gene editing based on Watson-Crick recognition without sequence restriction.

  4. Alkali-labile sites and post-irradiation effects in single-stranded DNA induced by H radicals

    International Nuclear Information System (INIS)

    Lafleur, M.V.M.; Heuvel, N. van; Woldhuis, J.; Loman, H.

    1978-01-01

    Single-stranded phiX174 DNA in aqueous solutions has been irradiated in the absence of oxygen, under conditions in which H radicals react with the DNA. It was shown that H radical reactions result in breaks, which contribute approximately 10 per cent inactivation. Further, two types of alkali-labile sites were formed. One was lethal and gave rise to single-strand breaks by alkali and was most probably identical with post-irradiation heat damage and contributed about 33 per cent to the inactivation mentioned above. The other consisted of non-lethal damage, partly dihydropyrimidine derivatives, and was converted to lethal damage by alkali. This followed from experiments in which the DNA was treated with osmium-tetroxide, which oxidized thymine to 5,6-dihydroxydihydrothymine. Treatment with alkali of this DNA gave the same temperature dependence as found for the non-lethal alkali-labile sites in irradiated DNA. A similar temperature dependence was found for dihydrothymine and irradiated pyrimidines with alkali. (author)

  5. Non-uniform binding of single-stranded DNA binding proteins to hybrids of single-stranded DNA and single-walled carbon nanotubes observed by atomic force microscopy in air and in liquid

    Energy Technology Data Exchange (ETDEWEB)

    Umemura, Kazuo, E-mail: meicun2006@163.com; Ishizaka, Kei; Nii, Daisuke; Izumi, Katsuki

    2016-12-01

    Highlights: • Conjugates of protein, DNA, and SWNTs were observed by AFM in liquid. • Non-uniform binding of proteins was visualized in liquid. • Thickness of DNA molecules on SWNT surfaces was well characterized in liquid. - Abstract: Using atomic force spectroscopy (AFM), we observed hybrids of single-stranded DNA (ssDNA) and single-walled carbon nanotubes (SWNTs) with or without protein molecules in air and in an aqueous solution. This is the first report of ssDNA–SWNT hybrids with proteins in solution analyzed by AFM. In the absence of protein, the height of the ssDNA–SWNT hybrids was 1.1 ± 0.3 nm and 2.4 ± 0.6 nm in air and liquid, respectively, suggesting that the ssDNA molecules adopted a flexible structure on the SWNT surface. In the presence of single-stranded DNA binding (SSB) proteins, the heights of the hybrids in air and liquid increased to 6.4 ± 3.1 nm and 10.0 ± 4.5 nm, respectively. The AFM images clearly showed binding of the SSB proteins to the ssDNA–SWNT hybrids. The morphology of the SSB–ssDNA–SWNT hybrids was non-uniform, particularly in aqueous solution. The variance of hybrid height was quantitatively estimated by cross-section analysis along the long-axis of each hybrid. The SSB–ssDNA–SWNT hybrids showed much larger variance than the ssDNA–SWNT hybrids.

  6. Base damage within single-strand DNA underlies in vivo hypermutability induced by a ubiquitous environmental agent.

    Directory of Open Access Journals (Sweden)

    Kin Chan

    Full Text Available Chromosomal DNA must be in single-strand form for important transactions such as replication, transcription, and recombination to occur. The single-strand DNA (ssDNA is more prone to damage than double-strand DNA (dsDNA, due to greater exposure of chemically reactive moieties in the nitrogenous bases. Thus, there can be agents that damage regions of ssDNA in vivo while being inert toward dsDNA. To assess the potential hazard posed by such agents, we devised an ssDNA-specific mutagenesis reporter system in budding yeast. The reporter strains bear the cdc13-1 temperature-sensitive mutation, such that shifting to 37°C results in telomere uncapping and ensuing 5' to 3' enzymatic resection. This exposes the reporter region, containing three closely-spaced reporter genes, as a long 3' ssDNA overhang. We validated the ability of the system to detect mutagenic damage within ssDNA by expressing a modified human single-strand specific cytosine deaminase, APOBEC3G. APOBEC3G induced a high density of substitutions at cytosines in the ssDNA overhang strand, resulting in frequent, simultaneous inactivation of two reporter genes. We then examined the mutagenicity of sulfites, a class of reactive sulfur oxides to which humans are exposed frequently via respiration and food intake. Sulfites, at a concentration similar to that found in some foods, induced a high density of mutations, almost always as substitutions at cytosines in the ssDNA overhang strand, resulting in simultaneous inactivation of at least two reporter genes. Furthermore, sulfites formed a long-lived adducted 2'-deoxyuracil intermediate in DNA that was resistant to excision by uracil-DNA N-glycosylase. This intermediate was bypassed by error-prone translesion DNA synthesis, frequently involving Pol ζ, during repair synthesis. Our results suggest that sulfite-induced lesions in DNA can be particularly deleterious, since cells might not possess the means to repair or bypass such lesions

  7. Three-dimensional deformation of orthodontic brackets.

    Science.gov (United States)

    Melenka, Garrett W; Nobes, David S; Major, Paul W; Carey, Jason P

    2013-01-01

    Braces are used by orthodontists to correct the misalignment of teeth in the mouth. Archwire rotation is a particular procedure used to correct tooth inclination. Wire rotation can result in deformation to the orthodontic brackets, and an orthodontic torque simulator has been designed to examine this wire-bracket interaction. An optical technique has been employed to measure the deformation due to size and geometric constraints of the orthodontic brackets. Images of orthodontic brackets are collected using a stereo microscope and two charge-coupled device cameras, and deformation of orthodontic brackets is measured using a three-dimensional digital image correlation technique. The three-dimensional deformation of orthodontic brackets will be evaluated. The repeatability of the three-dimensional digital image correlation measurement method was evaluated by performing 30 archwire rotation tests using the same bracket and archwire. Finally, five Damon 3MX and five In-Ovation R self-ligating brackets will be compared using this technique to demonstrate the effect of archwire rotation on bracket design.

  8. Functional characterization of a conserved archaeal viral operon revealing single-stranded DNA binding, annealing and nuclease activities

    DEFF Research Database (Denmark)

    Guo, Yang; Kragelund, Birthe Brandt; White, Malcolm F.

    2015-01-01

    encoding proteins of unknown function and forming an operon with ORF207 (gp19). SIRV2 gp17 was found to be a single-stranded DNA (ssDNA) binding protein different in structure from all previously characterized ssDNA binding proteins. Mutagenesis of a few conserved basic residues suggested a U......-shaped binding path for ssDNA. The recombinant gp18 showed an ssDNA annealing activity often associated with helicases and recombinases. To gain insight into the biological role of the entire operon, we characterized SIRV2 gp19 and showed it to possess a 5'→3' ssDNA exonuclease activity, in addition...... for rudiviruses and the close interaction among the ssDNA binding, annealing and nuclease proteins strongly point to a role of the gene operon in genome maturation and/or DNA recombination that may function in viral DNA replication/repair....

  9. Effect of nalidixic acid on repair of single-strand breaks in DNA induced by ionizing irradiation in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Francia, I [Debreceni Orvostudomanyi Egyetem (Hungary); Okos, A; Hernadi, F J [Institute of Pharmacology, Debrecen (Hungary)

    1978-09-30

    The incidence of DNA single-strand breaks induced by /sup 60/Co irradiation and their repair in E.coli K12 (AB 1157) rec/sup +/ cells were studied by the alkaline sucrose gradient sedimentation method described by McGrath and Williams. For the quantitative analysis of sedimentation profiles we used the s 1/2 values described by Veatch and Okada. The s 1/2 value of non-irradiated controls was 22.4, and after 20 krads irradiation it was found to be 11.7. A postirradiation incubation at 37 /sup 0/C for 60 min increasedthe s 1/2 value from 11.7 to 22.1. Nalidixic acid at low concentration (20-50 ..mu..g/ml) did not block, but at 100 ..mu..g/ml extensively inhibited the above repair process, exhibiting an s 1/2 value of 14.4.

  10. Packaging signals in two single-stranded RNA viruses imply a conserved assembly mechanism and geometry of the packaged genome.

    Science.gov (United States)

    Dykeman, Eric C; Stockley, Peter G; Twarock, Reidun

    2013-09-09

    The current paradigm for assembly of single-stranded RNA viruses is based on a mechanism involving non-sequence-specific packaging of genomic RNA driven by electrostatic interactions. Recent experiments, however, provide compelling evidence for sequence specificity in this process both in vitro and in vivo. The existence of multiple RNA packaging signals (PSs) within viral genomes has been proposed, which facilitates assembly by binding coat proteins in such a way that they promote the protein-protein contacts needed to build the capsid. The binding energy from these interactions enables the confinement or compaction of the genomic RNAs. Identifying the nature of such PSs is crucial for a full understanding of assembly, which is an as yet untapped potential drug target for this important class of pathogens. Here, for two related bacterial viruses, we determine the sequences and locations of their PSs using Hamiltonian paths, a concept from graph theory, in combination with bioinformatics and structural studies. Their PSs have a common secondary structure motif but distinct consensus sequences and positions within the respective genomes. Despite these differences, the distributions of PSs in both viruses imply defined conformations for the packaged RNA genomes in contact with the protein shell in the capsid, consistent with a recent asymmetric structure determination of the MS2 virion. The PS distributions identified moreover imply a preferred, evolutionarily conserved assembly pathway with respect to the RNA sequence with potentially profound implications for other single-stranded RNA viruses known to have RNA PSs, including many animal and human pathogens. Copyright © 2013 Elsevier Ltd. All rights reserved.

  11. Alkyladenine DNA glycosylase (AAG) localizes to mitochondria and interacts with mitochondrial single-stranded binding protein (mtSSB).

    Science.gov (United States)

    van Loon, Barbara; Samson, Leona D

    2013-03-01

    Due to a harsh environment mitochondrial genomes accumulate high levels of DNA damage, in particular oxidation, hydrolytic deamination, and alkylation adducts. While repair of alkylated bases in nuclear DNA has been explored in detail, much less is known about the repair of DNA alkylation damage in mitochondria. Alkyladenine DNA glycosylase (AAG) recognizes and removes numerous alkylated bases, but to date AAG has only been detected in the nucleus, even though mammalian mitochondria are known to repair DNA lesions that are specific substrates of AAG. Here we use immunofluorescence to show that AAG localizes to mitochondria, and we find that native AAG is present in purified human mitochondrial extracts, as well as that exposure to alkylating agent promotes AAG accumulation in the mitochondria. We identify mitochondrial single-stranded binding protein (mtSSB) as a novel interacting partner of AAG; interaction between mtSSB and AAG is direct and increases upon methyl methanesulfonate (MMS) treatment. The consequence of this interaction is specific inhibition of AAG glycosylase activity in the context of a single-stranded DNA (ssDNA), but not a double-stranded DNA (dsDNA) substrate. By inhibiting AAG-initiated processing of damaged bases, mtSSB potentially prevents formation of DNA breaks in ssDNA, ensuring that base removal primarily occurs in dsDNA. In summary, our findings suggest the existence of AAG-initiated BER in mitochondria and further support a role for mtSSB in DNA repair. Copyright © 2012. Published by Elsevier B.V.

  12. Charge Enhancement of Single-Stranded DNA in Negative Electrospray Ionization Using the Supercharging Reagent Meta-nitrobenzyl Alcohol

    Science.gov (United States)

    Brahim, Bessem; Alves, Sandra; Cole, Richard B.; Tabet, Jean-Claude

    2013-12-01

    Charge enhancement of single-stranded oligonucleotide ions in negative ESI mode is investigated. The employed reagent, meta-nitrobenzyl alcohol (m-NBA), was found to improve total signal intensity (Itot), increase the highest observed charge states (zhigh), and raise the average charge states (zavg) of all tested oligonucleotides analyzed in negative ESI. To quantify these increases, signal enhancement ratios (SER1%) and charge enhancement coefficients (CEC1%) were introduced. The SER1%, (defined as the quotient of total oligonucleotide ion abundances with 1 % m-NBA divided by total oligonucleotide abundance without m-NBA) was found to be greater than unity for every oligonucleotide tested. The CEC1% values (defined as the average charge state in the presence of 1 % m-NBA minus the average charge state in the absence of m-NBA) were found to be uniformly positive. Upon close inspection, the degree of charge enhancement for longer oligonucleotides was found to be dependent upon thymine density (i.e., the number and the location of phospho-thymidine units). A correlation between the charge enhancement induced by the presence of m-NBA and the apparent gas-phase acidity (largely determined by the sequence of thymine units but also by the presence of protons on other nucleobases) of multiply deprotonated oligonucleotide species, was thus established. Ammonium cations appeared to be directly involved in the m-NBA supercharging mechanism, and their role seems to be consistent with previously postulated ESI mechanisms describing desorption/ionization of single-stranded DNA into the gas phase.

  13. Single-Stranded Nucleic Acids Bind to the Tetramer Interface of SAMHD1 and Prevent Formation of the Catalytic Homotetramer.

    Science.gov (United States)

    Seamon, Kyle J; Bumpus, Namandjé N; Stivers, James T

    2016-11-08

    Sterile alpha motif and HD domain protein 1 (SAMHD1) is a unique enzyme that plays important roles in nucleic acid metabolism, viral restriction, and the pathogenesis of autoimmune diseases and cancer. Although much attention has been focused on its dNTP triphosphohydrolase activity in viral restriction and disease, SAMHD1 also binds to single-stranded RNA and DNA. Here we utilize a UV cross-linking method using 5-bromodeoxyuridine-substituted oligonucleotides coupled with high-resolution mass spectrometry to identify the binding site for single-stranded nucleic acids (ssNAs) on SAMHD1. Mapping cross-linked amino acids on the surface of existing crystal structures demonstrated that the ssNA binding site lies largely along the dimer-dimer interface, sterically blocking the formation of the homotetramer required for dNTPase activity. Surprisingly, the disordered C-terminus of SAMHD1 (residues 583-626) was also implicated in ssNA binding. An interaction between this region and ssNA was confirmed in binding studies using the purified SAMHD1 583-626 peptide. Despite a recent report that SAMHD1 possesses polyribonucleotide phosphorylase activity, we did not detect any such activity in the presence of inorganic phosphate, indicating that nucleic acid binding is unrelated to this proposed activity. These data suggest an antagonistic regulatory mechanism in which the mutually exclusive oligomeric state requirements for ssNA binding and dNTP hydrolase activity modulate these two functions of SAMHD1 within the cell.

  14. Structural features of single-stranded integron cassette attC sites and their role in strand selection.

    Directory of Open Access Journals (Sweden)

    Marie Bouvier

    2009-09-01

    Full Text Available We recently showed that cassette integration and deletion in integron platforms were occurring through unconventional site-specific recombination reactions involving only the bottom strand of attC sites. The lack of sequence conservation among attC sites led us to hypothesize that sequence-independent structural recognition determinants must exist within attC sites. The structural data obtained from a synaptic complex of the Vibrio cholerae integrase with the bottom strand of an attC site has shown the importance of extra helical bases (EHB inside the stem-loop structure formed from the bottom strand. Here, we systematically determined the contribution of three structural elements common to all known single-stranded attC site recombination substrates (the EHBs, the unpaired central spacer (UCS, and the variable terminal structure (VTS to strand choice and recombination. Their roles have been evaluated in vivo in the attIxattC reaction context using the suicide conjugation assay we previously developed, but also in an attCxattC reaction using a deletion assay. Conjugation was used to deliver the attC sites in single-stranded form. Our results show that strand choice is primarily directed by the first EHB, but the presence of the two other EHBs also serves to increase this strand selection. We found that the structure of the central spacer is essential to achieve high level recombination of the bottom strand, suggesting a dual role for this structure in active site exclusion and for hindering the reverse reaction after the first strand exchange. Moreover, we have shown that the VTS has apparently no role in strand selectivity.

  15. Bacillus subtilis single-stranded DNA-binding protein SsbA is phosphorylated at threonine 38 by the serine/threonine kinase YabT

    DEFF Research Database (Denmark)

    Derouiche, Abderahmane; Petranovic, Dina; Macek, Boris

    2016-01-01

    Background and purpose: Single-stranded DNA-binding proteins participate in all stages of DNA metabolism that involve single-stranded DNA, from replication, recombination, repair of DNA damage, to natural competence in species such as Bacillus subtilis. B. subtilis single-stranded DNA......-binding proteins have previously been found to be phosphorylated on tyrosine and arginine residues. While tyrosine phosphorylation was shown to enhance the DNA-binding properties of SsbA, arginine phosphorylation was not functionally characterized.Materials and methods: We used mass spectrometry analysis to detect...... phosphorylation of SsbA purified from B. subtilis cells. The detected phosphorylation site was assessed for its influence on DNA-binding in vitro, using electrophoretic mobility shift assays. The ability of B. subtilis serine/threonine kinases to phosphorylate SsbA was assessed using in vitro phosphorylation...

  16. Epidermal growth factor stimulating reparation of γ-ray-induced single-strand breaks predominantly in untranscribed DNA of HeLa cells

    International Nuclear Information System (INIS)

    Igusheva, O.A.; Bil'din, V.N.; Zhestyanikov, V.D.

    1994-01-01

    Considerable evidence suggest that genomic DNA undergoes reparation unevenly because of different transcription activities of its particular sequence. It is highly probably that transcriptional factors are necessary for postion stages of excision reparation and for reparation of single-strand DNA breaks caused by ionizing radiation. There is evidence suggesting that DNA lesions inflicted by γ-radiation is preferentially initiated in transcribed rather than in untranscribed DNA species. This paper looks at the relationship between stimulatory effect of epidermal growth factor (EGF) on reparation of single-strand DNA breaks and reparation of the damage done to active and inert fragments of chromatin. The results show that EGF stimulates reparation of single-strand DNA breaks induced by γ-radiation more effectively in untranscribed than in transcribed DNA. 13 refs., 1 fig., 1 tab

  17. Electropolishing-Orthodontic Office: A Simplified Approach

    Directory of Open Access Journals (Sweden)

    Sudhir Munjal

    2014-01-01

    Full Text Available Electropolishing plays an important role in dentistry by providing enhanced mechanical properties, better corrosion protection, physical appearance and ease of cleaning various metallic attachments. To achieve all these objectives, we present here a simple and economical way to fabricate a electropolisher which has wide applications in orthodontic office (recycle brackets, anneal retention wires, etc..

  18. Surgical orthodontics.

    Science.gov (United States)

    Strohl, Alexis M; Vitkus, Lauren

    2017-08-01

    The article reviews some commonly used orthodontic treatments as well as new strategies to assist in the correction of malocclusion. Many techniques are used in conjunction with surgical intervention and are a necessary compliment to orthognathic surgery. Basic knowledge of these practices will aid in the surgeon's ability to adequately treat the patient. Many orthodontists and surgeons are eliminating presurgical orthodontics to adopt a strategy of 'surgery first' orthodontics in orthognathic surgery. This has the benefit of immediate improvement in facial aesthetics and shorter treatment times. The advent of virtual surgical planning has helped facilitate the development of this new paradigm by making surgical planning faster and easier. Furthermore, using intraoperative surgical navigation is improving overall precision and outcomes. A variety of surgical and nonsurgical treatments may be employed in the treatment of malocclusion. It is important to be familiar with all options available and tailor the patient's treatment plan accordingly. Surgery-first orthodontics, intraoperative surgical navigation, virtual surgical planning, and 3D printing are evolving new techniques that are producing shorter treatment times and subsequently improving patient satisfaction without sacrificing long-term stability.

  19. Clinical bending of nickel titanium wires

    Directory of Open Access Journals (Sweden)

    Stephen Chain

    2015-01-01

    Full Text Available Since the evolution and the involvement of Nickel Titanium wires in the field of Orthodontics. The treatment plan has evolved with the use of low force Nickel Titanium wires. Because of their high springback, low stiffness, they are the key initial wires in leveling and alignment but have poor formability. Since poor formability limits its ability to create variable arch forms thus; limits the form of treatment. We have devised a method to bend the Nickel Titanium wires to help in our inventory but also customized the wire according to the treatment.

  20. Genetic and Biochemical Identification of a Novel Single-Stranded DNA-Binding Complex in Haloferax volcanii.

    Science.gov (United States)

    Stroud, Amy; Liddell, Susan; Allers, Thorsten

    2012-01-01

    Single-stranded DNA (ssDNA)-binding proteins play an essential role in DNA replication and repair. They use oligonucleotide/oligosaccharide-binding (OB)-folds, a five-stranded β-sheet coiled into a closed barrel, to bind to ssDNA thereby protecting and stabilizing the DNA. In eukaryotes the ssDNA-binding protein (SSB) is known as replication protein A (RPA) and consists of three distinct subunits that function as a heterotrimer. The bacterial homolog is termed SSB and functions as a homotetramer. In the archaeon Haloferax volcanii there are three genes encoding homologs of RPA. Two of the rpa genes (rpa1 and rpa3) exist in operons with a novel gene specific to Euryarchaeota; this gene encodes a protein that we have termed RPA-associated protein (rpap). The rpap genes encode proteins belonging to COG3390 group and feature OB-folds, suggesting that they might cooperate with RPA in binding to ssDNA. Our genetic analysis showed that rpa1 and rpa3 deletion mutants have differing phenotypes; only Δrpa3 strains are hypersensitive to DNA damaging agents. Deletion of the rpa3-associated gene rpap3 led to similar levels of DNA damage sensitivity, as did deletion of the rpa3 operon, suggesting that RPA3 and RPAP3 function in the same pathway. Protein pull-downs involving recombinant hexahistidine-tagged RPAs showed that RPA3 co-purifies with RPAP3, and RPA1 co-purifies with RPAP1. This indicates that the RPAs interact only with their respective associated proteins; this was corroborated by the inability to construct rpa1 rpap3 and rpa3 rpap1 double mutants. This is the first report investigating the individual function of the archaeal COG3390 RPA-associated proteins (RPAPs). We have shown genetically and biochemically that the RPAPs interact with their respective RPAs, and have uncovered a novel single-stranded DNA-binding complex that is unique to Euryarchaeota.

  1. Changes in the infrared microspectroscopic characteristics of DNA caused by cationic elements, different base richness and single-stranded form.

    Directory of Open Access Journals (Sweden)

    Maria Luiza S Mello

    Full Text Available BACKGROUND: The infrared (IR analysis of dried samples of DNA and DNA-polypeptide complexes is still scarce. Here we have studied the FT-IR profiles of these components to further the understanding of the FT-IR signatures of chromatin and cell nuclei. METHODOLOGY/PRINCIPAL FINDINGS: Calf thymus and salmon testis DNA, and complexes of histone H1, protamine, poly-L-lysine and poly-L-arginine (histone-mimic macromolecules with DNA were analyzed in an IR microspectroscope equipped with an attenuated total reflection diamond objective and Grams software. Conditions including polypeptides bound to the DNA, DNA base composition, and single-stranded form were found to differently affect the vibrational characteristics of the chemical groups (especially, PO(2(- in the nucleic acid. The antisymmetric stretching (ν(as of the DNA PO(2(- was greater than the symmetric stretching (ν(s of these groups and increased in the polypeptide-DNA complexes. A shift of the ν(as of the DNA PO(2(- to a lower frequency and an increased intensity of this vibration were induced especially by lysine-rich histones. Lysine richness additionally contributed to an increase in the vibrational stretching of the amide I group. Even in simple molecules such as inorganic phosphates, the vibrational characteristics of the phosphate anions were differently affected by different cations. As a result of the optimization of the DNA conformation by binding to arginine-rich polypeptides, enhancements of the vibrational characteristics in the FT-IR fingerprint could be detected. Although different profiles were obtained for the DNA with different base compositions, this situation was no longer verified in the polypeptide-DNA complexes and most likely in isolated chromatin or cell nuclei. However, the ν(as PO(2(-/ν(s PO(2(- ratio could discriminate DNA with different base compositions and DNA in a single-stranded form. CONCLUSIONS/SIGNIFICANCE: FT-IR spectral profiles are a valuable tool

  2. Initiation and termination of the bacteriophage phi X174 rolling circle DNA replication in vivo: packaging of plasmid single-stranded DNA into bacteriophage phi X174 coats

    NARCIS (Netherlands)

    van der Ende, A.; Teertstra, R.; Weisbeek, P. J.

    1982-01-01

    The bacteriophage phi X174 viral (+) origin when inserted in a plasmid can interact in vivo with the A protein produced by infecting phi X174 phages. A consequence of this interaction is packaging of single-stranded plasmid DNA into preformed phage coats resulting in infective particles (1). This

  3. Mechanism of replication of ultraviolet-irradiated single-stranded DNA by DNA polymerase III holoenzyme of Escherichia coli. Implications for SOS mutagenesis

    International Nuclear Information System (INIS)

    Livneh, Z.

    1986-01-01

    Replication of UV-irradiated oligodeoxynucleotide-primed single-stranded phi X174 DNA with Escherichia coli DNA polymerase III holoenzyme in the presence of single-stranded DNA-binding protein was investigated. The extent of initiation of replication on the primed single-stranded DNA was not altered by the presence of UV-induced lesions in the DNA. The elongation step exhibited similar kinetics when either unirradiated or UV-irradiated templates were used. Inhibition of the 3'----5' proofreading exonucleolytic activity of the polymerase by dGMP or by a mutD mutation did not increase bypass of pyrimidine photodimers, and neither did purified RecA protein influence the extent of photodimer bypass as judged by the fraction of full length DNA synthesized. Single-stranded DNA-binding protein stimulated bypass since in its absence the fraction of full length DNA decreased 5-fold. Termination of replication at putative pyrimidine dimers involved dissociation of the polymerase from the DNA, which could then reinitiate replication at other available primer templates. Based on these observations a model for SOS-induced UV mutagenesis is proposed

  4. Molecular dosimetry of DNA damage caused by alkylation. I. Single-strand breaks induced by ethylating agents in cultured mammalian cells in relation to survival

    NARCIS (Netherlands)

    Abbondandolo, A.; Dogliotti, E.; Lohman, P.H.M.; Berends, F.

    1982-01-01

    Cultured Chinese hamster ovary cells were treated with ethylating agents. DNA lesions giving rise to single-strand breaks (ssb) or alkali-labile sites were measured by centrifugation in alkaline sucrose gradients after lysis in alkali. 4 agents with different tendencies to ethylate preferentially

  5. Induction of single-strand DNA breaks in human cells by H2O2 formed in near-uv (black light)-irradiated medium

    International Nuclear Information System (INIS)

    Wang, R.J.; Ananthaswamy, H.N.; Nixon, B.T.; Hartman, P.S.; Eisenstark, A.

    1980-01-01

    When Dulbecco's modified Eagle's medium (depleted of phenol red) was irradiated for up to 3 h by 4 to 5 W/m 2 black light, hydrogen peroxide (H 2 O 2 ) was produced. Generation of H 2 O 2 resulted from riboflavin-sensitized photooxidation of tryptophan and tyrosine. Reagent H 2 O 2 , or hydrogen peroxide generated in black light-exposed aqueous solutions containing riboflavin and tryptophan, induced 2 x 10 4 single-strand breaks per 10 16 daltons of DNA in intact, physiologically viable human D98/AH 2 cells. Concomitant with the single-strand breaks in the cells was loss of cellular reproductive viability. Two classes of photoproducts were identified: H 2 O 2 and non-H 2 O 2 . The H 2 O 2 component of the photoproducts was responsible for all the single-strand break induction but for only partial loss of reproductive viability. The non-H 2 O 2 photoproducts, accountable for the remainder of cell lethality, caused no single-strand breaks

  6. Micronuclei, DNA single-strand breaks and DNA-repair activity in mice exposed to 1,3-butadiene by inhalation

    Czech Academy of Sciences Publication Activity Database

    Vodička, Pavel; Štětina, R.; Šmerák, P.; Vodičková, Ludmila; Naccarati, Alessio; Bárta, I.; Hemminki, K.

    2006-01-01

    Roč. 608, - (2006), s. 49-57 ISSN 1383-5718 R&D Projects: GA ČR(CZ) GA310/01/0802 Institutional research plan: CEZ:AV0Z50390512 Keywords : Single-strand DNA breaks * Micronucleus formation * DNA-repair activity Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.122, year: 2006

  7. Effects of Fluoride on NiTi Orthodontic Archwires: An X-ray Diffraction Study

    Directory of Open Access Journals (Sweden)

    Sumit Kumar Yadav

    2013-01-01

    Results: Unloading force values of NiTi orthodontic wires were significantly decreased after exposure to both fluoride solutions (p < 0.001. Corrosive changes in surface topography were observed for both fluoride solutions. Wires exposed to acidic fluoride appeared as more severely affected. X-ray diffraction analysis showed no change in crystal lattice of NiTi wires in both solutions. Conclusion: The results suggest that using topical fluoride agents with NiTi wire could decrease the functional unloading mechanical properties of the wire and contribute to prolonged orthodontic treatment.

  8. Strand displacement by DNA polymerase III occurs through a tau-psi-chi link to single-stranded DNA-binding protein coating the lagging strand template.

    Science.gov (United States)

    Yuan, Quan; McHenry, Charles S

    2009-11-13

    In addition to the well characterized processive replication reaction catalyzed by the DNA polymerase III holoenzyme on single-stranded DNA templates, the enzyme possesses an intrinsic strand displacement activity on flapped templates. The strand displacement activity is distinguished from the single-stranded DNA-templated reaction by a high dependence upon single-stranded DNA binding protein and an inability of gamma-complex to support the reaction in the absence of tau. However, if gamma-complex is present to load beta(2), a truncated tau protein containing only domains III-V will suffice. This truncated protein is sufficient to bind both the alpha subunit of DNA polymerase (Pol) III and chipsi. This is reminiscent of the minimal requirements for Pol III to replicate short single-stranded DNA-binding protein (SSB)-coated templates where tau is only required to serve as a scaffold to hold Pol III and chi in the same complex (Glover, B., and McHenry, C. (1998) J. Biol. Chem. 273, 23476-23484). We propose a model in which strand displacement by DNA polymerase III holoenzyme depends upon a Pol III-tau-psi-chi-SSB binding network, where SSB is bound to the displaced strand, stabilizing the Pol III-template interaction. The same interaction network is probably important for stabilizing the leading strand polymerase interactions with authentic replication forks. The specificity constant (k(cat)/K(m)) for the strand displacement reaction is approximately 300-fold less favorable than reactions on single-stranded templates and proceeds with a slower rate (150 nucleotides/s) and only moderate processivity (approximately 300 nucleotides). PriA, the initiator of replication restart on collapsed or misassembled replication forks, blocks the strand displacement reaction, even if added to an ongoing reaction.

  9. Craniofacial orthodontics and postgraduate orthodontic training in ...

    African Journals Online (AJOL)

    The least clinical experience was recorded in pre-bone graft orthodontics (7.4%) and orthodontic preparation for orthognathic surgery (5.5%). Some of the challenges highlighted by the residents were low patients turn out for orthodontic care and the absence of multidisciplinary treatment for craniofacial patients in their ...

  10. Single-strand DNA-binding protein SSB1 facilitates TERT recruitment to telomeres and maintains telomere G-overhangs.

    Science.gov (United States)

    Pandita, Raj K; Chow, Tracy T; Udayakumar, Durga; Bain, Amanda L; Cubeddu, Liza; Hunt, Clayton R; Shi, Wei; Horikoshi, Nobuo; Zhao, Yong; Wright, Woodring E; Khanna, Kum Kum; Shay, Jerry W; Pandita, Tej K

    2015-03-01

    Proliferating mammalian stem and cancer cells express telomerase [telomerase reverse transcriptase (TERT)] in an effort to extend chromosomal G-overhangs and maintain telomere ends. Telomerase-expressing cells also have higher levels of the single-stranded DNA-binding protein SSB1, which has a critical role in DNA double-strand break (DSB) repair. Here, we report that SSB1 binds specifically to G-strand telomeric DNA in vitro and associates with telomeres in vivo. SSB1 interacts with the TERT catalytic subunit and regulates its interaction with telomeres. Deletion of SSB1 reduces TERT interaction with telomeres and leads to G-overhang loss. Although SSB1 is recruited to DSB sites, we found no corresponding change in TERT levels at these sites, implying that SSB1-TERT interaction relies upon a specific chromatin structure or context. Our findings offer an explanation for how telomerase is recruited to telomeres to facilitate G-strand DNA extension, a critical step in maintaining telomere ends and cell viability in all cancer cells. Cancer Res; 75(5); 858-69. ©2015 AACR. ©2015 American Association for Cancer Research.

  11. Single-strand DNA binding protein SSB1 facilitates TERT recruitment to telomeres and maintains telomere G-overhangs

    Science.gov (United States)

    Pandita, Raj K.; Chow, Tracy T.; Udayakumar, Durga; Bain, Amanda L.; Cubeddu, Liza; Hunt, Clayton R.; Shi, Wei; Horikoshi, Nobuo; Zhao, Yong; Wright, Woodring E.; Khanna, Kum Kum; Shay, Jerry W.; Pandita, Tej K.

    2015-01-01

    Proliferating mammalian stem and cancer cells express telomerase (TERT) in an effort to extend chromosomal G-overhangs and maintain telomere ends. Telomerase-expressing cells also have higher levels of the single-stranded DNA binding protein SSB1, which has a critical role in DNA double-strand break repair. Here we report that SSB1 binds specifically to G-strand telomeric DNA in vitro and associates with telomeres in vivo. SSB1 interacted with the TERT catalytic subunit and regulates its interaction with telomeres. Deletion of SSB1 reduced TERT interaction with telomeres and lead to G-overhang loss. While SSB1 was recruited to DSB sites, we found no corresponding change in TERT levels at these sites, implying that SSB1-TERT interaction relied upon a specific chromatin structure or context. Our findings offer an explanation for how telomerase is recruited to telomeres to facilitate G-strand DNA extension, a critical step in maintaining telomere ends and cell viability in all cancer cells. PMID:25589350

  12. Structural Basis of Mec1-Ddc2-RPA Assembly and Activation on Single-Stranded DNA at Sites of Damage.

    Science.gov (United States)

    Deshpande, Ishan; Seeber, Andrew; Shimada, Kenji; Keusch, Jeremy J; Gut, Heinz; Gasser, Susan M

    2017-10-19

    Mec1-Ddc2 (ATR-ATRIP) is a key DNA-damage-sensing kinase that is recruited through the single-stranded (ss) DNA-binding replication protein A (RPA) to initiate the DNA damage checkpoint response. Activation of ATR-ATRIP in the absence of DNA damage is lethal. Therefore, it is important that damage-specific recruitment precedes kinase activation, which is achieved at least in part by Mec1-Ddc2 homodimerization. Here, we report a structural, biochemical, and functional characterization of the yeast Mec1-Ddc2-RPA assembly. High-resolution co-crystal structures of Ddc2-Rfa1 and Ddc2-Rfa1-t11 (K45E mutant) N termini and of the Ddc2 coiled-coil domain (CCD) provide insight into Mec1-Ddc2 homodimerization and damage-site targeting. Based on our structural and functional findings, we present a Mec1-Ddc2-RPA-ssDNA composite structural model. By way of validation, we show that RPA-dependent recruitment of Mec1-Ddc2 is crucial for maintaining its homodimeric state at ssDNA and that Ddc2's recruitment domain and CCD are important for Mec1-dependent survival of UV-light-induced DNA damage. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. RPA-coated single-stranded DNA as a platform for post-translational modifications in the DNA damage response.

    Science.gov (United States)

    Maréchal, Alexandre; Zou, Lee

    2015-01-01

    The Replication Protein A (RPA) complex is an essential regulator of eukaryotic DNA metabolism. RPA avidly binds to single-stranded DNA (ssDNA) through multiple oligonucleotide/oligosaccharide-binding folds and coordinates the recruitment and exchange of genome maintenance factors to regulate DNA replication, recombination and repair. The RPA-ssDNA platform also constitutes a key physiological signal which activates the master ATR kinase to protect and repair stalled or collapsed replication forks during replication stress. In recent years, the RPA complex has emerged as a key target and an important regulator of post-translational modifications in response to DNA damage, which is critical for its genome guardian functions. Phosphorylation and SUMOylation of the RPA complex, and more recently RPA-regulated ubiquitination, have all been shown to control specific aspects of DNA damage signaling and repair by modulating the interactions between RPA and its partners. Here, we review our current understanding of the critical functions of the RPA-ssDNA platform in the maintenance of genome stability and its regulation through an elaborate network of covalent modifications.

  14. PARP inhibition versus PARP-1 silencing: different outcomes in terms of single-strand break repair and radiation susceptibility

    International Nuclear Information System (INIS)

    Godon, C.; Cordelieres, F.P.; Giocanti, N.; Megnin-Chanet, F.; Hall, J.; Favaudon, V.; Godon, C.; Giocanti, N.; Megnin-Chanet, F.; Hall, J.; Favaudon, V.; Cordelieres, F.P.; Cordelieres, F.P.; Biard, D.

    2008-01-01

    The consequences of PARP-1 disruption or inhibition on DNA single-strand break repair (SSBR) and radio-induced lethality were determined in synchronized, iso-genic HeLa cells stably silenced or not for poly(ADP-ribose) polymerase-1 (PARP-1) (PARP-1(KD)) or XRCC1 (XRCC1(KD)). PARP-1 inhibition prevented XRCC1-YFP recruitment at sites of 405 nm laser micro irradiation, slowed SSBR 10-fold and triggered the accumulation of large persistent foci of GFP-PARP-1 and GFP-PCNA at photo damaged sites. These aggregates are presumed to hinder the recruitment of other effectors of the base excision repair (BER) pathway.PARP-1 silencing also prevented XRCC1-YFP recruitment but did not lengthen the lifetime of GFP-PCNA foci. Moreover, PARP-1(KD) and XRCC1(KD) cells in S phase completed SSBR as rapidly as controls, while SSBR was delayed in G1. Taken together, the data demonstrate that a PARP-1- and XRCC1-independent SSBR pathway operates when the short patch repair branch of the BER is deficient. Long patch repair is the likely mechanism, as GFP-PCNA recruitment at photo-damaged sites was normal in PARP-1(KD) cells. PARP-1 silencing elicited hyper-radiosensitivity, while radiosensitization by a PARP inhibitor reportedly occurs only in those cells treated in S phase. PARP-1 inhibition and deletion thus have different outcomes in terms of SSBR and radiosensitivity. (authors)

  15. Distinct spatio temporal patterns and PARP dependence of XRCC1 recruitment to single-strand break and base excision repair

    International Nuclear Information System (INIS)

    Campalans, Anna; Kortulewski, Thierry; Amouroux, Rachel; Radicella, J. Pablo; Menoni, Herve; Vermeulen, Wim

    2013-01-01

    Single-strand break repair (SSBR) and base excision repair (BER) of modified bases and abasic sites share several players. Among them is XRCC1, an essential scaffold protein with no enzymatic activity, required for the coordination of both pathways. XRCC1 is recruited to SSBR by PARP-1, responsible for the initial recognition of the break. The recruitment of XRCC1 to BER is still poorly understood. Here we show by using both local and global induction of oxidative DNA base damage that XRCC1 participation in BER complexes can be distinguished from that in SSBR by several criteria. We show first that XRCC1 recruitment to BER is independent of PARP. Second, unlike SSBR complexes that are assembled within minutes after global damage induction, XRCC1 is detected later in BER patches, with kinetics consistent with the repair of oxidized bases. Third, while XRCC1-containing foci associated with SSBR are formed both in eu- and heterochromatin domains, BER complexes are assembled in patches that are essentially excluded from heterochromatin and where the oxidized bases are detected. (authors)

  16. Assembly of presynaptic filaments. Factors affecting the assembly of RecA protein onto single-stranded DNA

    DEFF Research Database (Denmark)

    Thresher, RJ; Christiansen, Gunna; Griffith, JD

    1988-01-01

    We have previously shown that the assembly of RecA protein onto single-stranded DNA (ssDNA) facilitated by SSB protein occurs in three steps: (1) rapid binding of SSB protein to the ssDNA; (2) nucleation of RecA protein onto this template; and (3) co-operative polymerization of additional Rec......M in the presence of 12 mM-Mg2+), and relatively low concentrations of SSB protein (1 monomer per 18 nucleotides). Assembly was depressed threefold when SSB protein was added to one monomer per nine nucleotides. These effects appeared to be exerted at the nucleation step. Following nucleation, RecA protein...... assembled onto ssDNA at net rates that varied from 250 to 900 RecA protein monomers per minute, with the rate inversely related to the concentration of SSB protein. Combined sucrose sedimentation and electron microscope analysis established that SSB protein was displaced from the ssDNA during RecA protein...

  17. Electron microscopic comparison of the sequences of single-stranded genomes of mammalian parvoviruses by heteroduplex mapping

    Energy Technology Data Exchange (ETDEWEB)

    Banerjee, P.T.; Olson, W.H.; Allison, D.P.; Bates, R.C.; Snyder, C.E.; Mitra, S.

    1983-01-01

    The sequence homologies among the linear single-stranded genomes of several mammalian parvoviruses have been studied by electron microscopic analysis of tthe heteroduplexes produced by reannealing the complementary strands of their DNAs. The genomes of Kilham rat virus, H-1, minute virus of ice and LuIII, which are antigenically distinct non-defective parvoviruses, have considerable homology: about 70% of their sequences are conserved. The homologous regions map at similar locations in the left halves (from the 3' ends) of the genomes. No sequence homology, however, is observed between the DNAs of these nondefective parvoviruses and that of bovine parvovirus, another non-defective virus, or that of defective adenoassociated virus, nor between the genomes of bovine parvovirus and adenoassociated virus. This suggests that only very short, if any, homologous regions are present. From these results, an evolutionary relationship among Kilham rat virus, H-1, minute virus of mice and LuIII is predicted. It is interesting to note that, although LuIII was originally isolated from a human cell line and is specific for human cells in vitro, its genome has sequences in common only with the rodent viruses Kilham rat virus, minute virus of mice and H-1, and not with the other two mammalian parvoviruses tested.

  18. Overproduction of single-stranded-DNA-binding protein specifically inhibits recombination of UV-irradiated bacteriophage DNA in Escherichia coli

    International Nuclear Information System (INIS)

    Moreau, P.L.

    1988-01-01

    Overproduction of single-stranded DNA (ssDNA)-binding protein (SSB) in uvr Escherichia coli mutants results in a wide range of altered phenotypes. (i) Cell survival after UV irradiation is decreased; (ii) expression of the recA-lexA regulon is slightly reduced after UV irradiation, whereas it is increased without irradiation; and (iii) recombination of UV-damaged lambda DNA is inhibited, whereas recombination of nonirradiated DNA is unaffected. These results are consistent with the idea that in UV-damaged bacteria, SSB is first required to allow the formation of short complexes of RecA protein and ssDNA that mediate cleavage of the LexA protein. However, in a second stage, SSB should be displaced from ssDNA to permit the production of longer RecA-ssDNA nucleoprotein filaments that are required for strand pairing and, hence, recombinational repair. Since bacteria overproducing SSB appear identical in physiological respects to recF mutant bacteria, it is suggested that the RecF protein (alone or with other proteins of the RecF pathway) may help RecA protein to release SSB from ssDNA

  19. Structure-spectrophotometric selectivity relationship in interactions of quercetin related flavonoids with double stranded and single stranded RNA

    Science.gov (United States)

    Piantanida, Ivo; Mašić, Lozika; Rusak, Gordana

    2009-04-01

    Interactions of five flavonoids with dsRNA and single stranded ssRNA were studied by UV/vis titrations. The results obtained supported the intercalative binding mode as a dominant interaction of studied flavonoids with dsRNA as well as major interaction with ssRNA. Furthermore, changes of the UV/vis spectra of flavonoids induced by addition of poly G or poly C, respectively, are significantly stronger than changes induced by double stranded poly G-poly C, pointing to essential role of the free poly G or poly C sequence (not hydrogen bonded in double helix). Exclusively poly G caused significant batochromic shift of the UV/vis maxima of all studied flavonoids, whereby the intensity of batochromic shift is nicely correlated to the number of OH groups of flavonoid. Unlikely to poly G, addition of poly A and poly U induced measurable changes only in the UV/vis spectra of flavonoids characterised by no OH (galangin) or three OH groups (myricetin) on the phenyl part of the molecule. Consequently, flavonoids with one- or two-OH groups on the phenyl part of the molecule (luteolin, fisetin, kaempferol) specifically differentiate between poly A, poly U (negligible changes in the UV/Vis spectra) and poly G (strong changes in the UV/Vis spectra) as well as poly C (moderate changes in the UV/Vis spectra).

  20. Interaction with Single-stranded DNA-binding Protein Stimulates Escherichia coli Ribonuclease HI Enzymatic Activity

    Energy Technology Data Exchange (ETDEWEB)

    Petzold, Christine; Marceau, Aimee H.; Miller, Katherine H.; Marqusee, Susan; Keck, James L. (UW-MED); (UCB)

    2015-04-22

    Single-stranded (ss) DNA-binding proteins (SSBs) bind and protect ssDNA intermediates formed during replication, recombination, and repair reactions. SSBs also directly interact with many different genome maintenance proteins to stimulate their enzymatic activities and/or mediate their proper cellular localization. We have identified an interaction formed between Escherichia coli SSB and ribonuclease HI (RNase HI), an enzyme that hydrolyzes RNA in RNA/DNA hybrids. The RNase HI·SSB complex forms by RNase HI binding the intrinsically disordered C terminus of SSB (SSB-Ct), a mode of interaction that is shared among all SSB interaction partners examined to date. Residues that comprise the SSB-Ct binding site are conserved among bacterial RNase HI enzymes, suggesting that RNase HI·SSB complexes are present in many bacterial species and that retaining the interaction is important for its cellular function. A steady-state kinetic analysis shows that interaction with SSB stimulates RNase HI activity by lowering the reaction Km. SSB or RNase HI protein variants that disrupt complex formation nullify this effect. Collectively our findings identify a direct RNase HI/SSB interaction that could play a role in targeting RNase HI activity to RNA/DNA hybrid substrates within the genome.

  1. Change of conformation and internal dynamics of supercoiled DNA upon binding of Escherichia coli single-strand binding protein

    International Nuclear Information System (INIS)

    Langowski, J.; Benight, A.S.; Fujimoto, B.S.; Schurr, J.M.; Schomburg, U.

    1985-01-01

    The influence of Escherichia coli single-strand binding (SSB) protein on the conformation and internal dynamics of pBR322 and pUC8 supercoiled DNAs has been investigated by using dynamic light scattering at 632.8 and 351.1 nm and time-resolved fluorescence polarization anisotropy of intercalated ethidium. SSB protein binds to both DNAs up to a stoichiometry that is sufficient to almost completely relax the superhelical turns. Upon saturation binding, the translational diffusion coefficients (D 0 ) of both DNAs decrease by approximately 20%. Apparent diffusion coefficients (D/sub app/) obtained from dynamic light scattering display the well-known increase with K 2 (K = scattering vector), leveling off toward a plateau value (D/sub plat/) at high K 2 . For both DNAs, the difference D/sub plat/ - D 0 increases upon relaxation of supercoils by SSB protein, which indicates a corresponding enhancement of the subunit mobilities in internal motions. Fluorescence polarization anisotropy measurements on free and complexed pBR322 DNA indicate a (predominantly) uniform torsional rigidity for the saturated DNA/SSB protein complex that is significantly reduced compared to the free DNA. These observations are all consistent with the notion that binding of SSB protein is accompanied by a gradual loss of supercoils and saturates when the superhelical twist is largely removed

  2. Fanconi anemia complementation group A (FANCA) protein has intrinsic affinity for nucleic acids with preference for single-stranded forms.

    Science.gov (United States)

    Yuan, Fenghua; Qian, Liangyue; Zhao, Xinliang; Liu, Jesse Y; Song, Limin; D'Urso, Gennaro; Jain, Chaitanya; Zhang, Yanbin

    2012-02-10

    The Fanconi anemia complementation group A (FANCA) gene is one of 15 disease-causing genes and has been found to be mutated in ∼60% of Fanconi anemia patients. Using purified protein, we report that human FANCA has intrinsic affinity for nucleic acids. FANCA binds to both single-stranded (ssDNA) and double-stranded (dsDNA) DNAs; however, its affinity for ssDNA is significantly higher than for dsDNA in an electrophoretic mobility shift assay. FANCA also binds to RNA with an intriguingly higher affinity than its DNA counterpart. FANCA requires a certain length of nucleic acids for optimal binding. Using DNA and RNA ladders, we determined that the minimum number of nucleotides required for FANCA recognition is ∼30 for both DNA and RNA. By testing the affinity between FANCA and a variety of DNA structures, we found that a 5'-flap or 5'-tail on DNA facilitates its interaction with FANCA. A patient-derived FANCA truncation mutant (Q772X) has diminished affinity for both DNA and RNA. In contrast, the complementing C-terminal fragment of Q772X, C772-1455, retains the differentiated nucleic acid-binding activity (RNA > ssDNA > dsDNA), indicating that the nucleic acid-binding domain of FANCA is located primarily at its C terminus, where most disease-causing mutations are found.

  3. Fanconi Anemia Complementation Group A (FANCA) Protein Has Intrinsic Affinity for Nucleic Acids with Preference for Single-stranded Forms*

    Science.gov (United States)

    Yuan, Fenghua; Qian, Liangyue; Zhao, Xinliang; Liu, Jesse Y.; Song, Limin; D'Urso, Gennaro; Jain, Chaitanya; Zhang, Yanbin

    2012-01-01

    The Fanconi anemia complementation group A (FANCA) gene is one of 15 disease-causing genes and has been found to be mutated in ∼60% of Fanconi anemia patients. Using purified protein, we report that human FANCA has intrinsic affinity for nucleic acids. FANCA binds to both single-stranded (ssDNA) and double-stranded (dsDNA) DNAs; however, its affinity for ssDNA is significantly higher than for dsDNA in an electrophoretic mobility shift assay. FANCA also binds to RNA with an intriguingly higher affinity than its DNA counterpart. FANCA requires a certain length of nucleic acids for optimal binding. Using DNA and RNA ladders, we determined that the minimum number of nucleotides required for FANCA recognition is ∼30 for both DNA and RNA. By testing the affinity between FANCA and a variety of DNA structures, we found that a 5′-flap or 5′-tail on DNA facilitates its interaction with FANCA. A patient-derived FANCA truncation mutant (Q772X) has diminished affinity for both DNA and RNA. In contrast, the complementing C-terminal fragment of Q772X, C772–1455, retains the differentiated nucleic acid-binding activity (RNA > ssDNA > dsDNA), indicating that the nucleic acid-binding domain of FANCA is located primarily at its C terminus, where most disease-causing mutations are found. PMID:22194614

  4. A Novel Single-Strand RNAi Therapeutic Agent Targeting the (Pro)renin Receptor Suppresses Ocular Inflammation.

    Science.gov (United States)

    Kanda, Atsuhiro; Ishizuka, Erdal Tan; Shibata, Atsushi; Matsumoto, Takahiro; Toyofuku, Hidekazu; Noda, Kousuke; Namba, Kenichi; Ishida, Susumu

    2017-06-16

    The receptor-associated prorenin system (RAPS) refers to the pathogenic mechanism whereby prorenin binding to the (pro)renin receptor [(P)RR] dually activates the tissue renin-angiotensin system (RAS) and RAS-independent intracellular signaling. Here we revealed significant upregulation of prorenin and soluble (P)RR levels in the vitreous fluid of patients with uveitis compared to non-inflammatory controls, together with a positive correlation between these RAPS components and monocyte chemotactic protein-1 among several upregulated cytokines. Moreover, we developed a novel single-strand RNAi agent, proline-modified short hairpin RNA directed against human and mouse (P)RR [(P)RR-PshRNA], and we determined its safety and efficacy in vitro and in vivo. Application of (P)RR-PshRNA in mice caused significant amelioration of acute (uveitic) and chronic (diabetic) models of ocular inflammation with no apparent adverse effects. Our findings demonstrate the significant implication of RAPS in the pathogenesis of human uveitis and the potential usefulness of (P)RR-PshRNA as a therapeutic agent to reduce ocular inflammation. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  5. DFT investigations of phosphotriesters hydrolysis in aqueous solution: a model for DNA single strand scission induced by N-nitrosoureas.

    Science.gov (United States)

    Liu, Tingting; Zhao, Lijiao; Zhong, Rugang

    2013-02-01

    DNA phosphotriester adducts are common alkylation products of DNA phosphodiester moiety induced by N-nitrosoureas. The 2-hydroxyethyl phosphotriester was reported to hydrolyze more rapidly than other alkyl phosphotriesters both in neutral and in alkaline conditions, which can cause DNA single strand scission. In this work, DFT calculations have been employed to map out the four lowest activation free-energy profiles for neutral and alkaline hydrolysis of triethyl phosphate (TEP) and diethyl 2-hydroxyethyl phosphate (DEHEP). All the hydrolysis pathways were illuminated to be stepwise involving an acyclic or cyclic phosphorane intermediate for TEP or DEHEP, respectively. The rate-limiting step for all the hydrolysis reactions was found to be the formation of phosphorane intermediate, with the exception of DEHEP hydrolysis in alkaline conditions that the decomposition process turned out to be the rate-limiting step, owing to the extraordinary low formation barrier of cyclic phosphorane intermediate catalyzed by hydroxide. The rate-limiting barriers obtained for the four reactions are all consistent with the available experimental information concerning the corresponding hydrolysis reactions of phosphotriesters. Our calculations performed on the phosphate triesters hydrolysis predict that the lower formation barriers of cyclic phosphorane intermediates compared to its acyclic counter-part should be the dominant factor governing the hydrolysis rate enhancement of DEHEP relative to TEP both in neutral and in alkaline conditions.

  6. Interaction of bacteriophage T4 and T7 single-stranded DNA-binding proteins with DNA

    International Nuclear Information System (INIS)

    Shokri, Leila; Williams, Mark C; Rouzina, Ioulia

    2009-01-01

    Bacteriophages T4 and T7 are well-studied model replication systems, which have allowed researchers to determine the roles of many proteins central to DNA replication, recombination and repair. Here we summarize and discuss the results from two recently developed single-molecule methods to determine the salt-dependent DNA-binding kinetics and thermodynamics of the single-stranded DNA (ssDNA)-binding proteins (SSBs) from these systems. We use these methods to characterize both the equilibrium double-stranded DNA (dsDNA) and ssDNA binding of the SSBs T4 gene 32 protein (gp32) and T7 gene 2.5 protein (gp2.5). Despite the overall two-orders-of-magnitude weaker binding of gp2.5 to both forms of DNA, we find that both proteins exhibit four-orders-of-magnitude preferential binding to ssDNA relative to dsDNA. This strong preferential ssDNA binding as well as the weak dsDNA binding is essential for the ability of both proteins to search dsDNA in one dimension to find available ssDNA-binding sites at the replication fork

  7. The single-strand DNA binding activity of human PC4 preventsmutagenesis and killing by oxidative DNA damage

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jen-Yeu; Sarker, Altaf Hossain; Cooper, Priscilla K.; Volkert, Michael R.

    2004-02-01

    Human positive cofactor 4 (PC4) is a transcriptional coactivator with a highly conserved single-strand DNA (ssDNA) binding domain of unknown function. We identified PC4 as a suppressor of the oxidative mutator phenotype of the Escherichia coli fpg mutY mutant and demonstrate that this suppression requires its ssDNA binding activity. Yeast mutants lacking their PC4 ortholog Sub1 are sensitive to hydrogen peroxide and exhibit spontaneous and peroxide induced hypermutability. PC4 expression suppresses the peroxide sensitivity of the yeast sub l{Delta} mutant, suggesting that the human protein has a similar function. A role for yeast and human proteins in DNA repair is suggested by the demonstration that Sub1 acts in a peroxide-resistance pathway involving Rad2 and by the physical interaction of PC4 with the human Rad2 homolog XPG. We show XPG recruits PC4 to a bubble-containing DNA substrate with resulting displacement of XPG and formation of a PC4-DNA complex. We discuss the possible requirement for PC4 in either global or transcription-coupled repair of oxidative DNA damage to mediate the release of XPG bound to its substrate.

  8. DNA single-strand breaks during repair of uv damage in human fibroblasts and abnormalities of repair in xeroderma pigmentosum

    International Nuclear Information System (INIS)

    Fornace, A.J. Jr.; Kohn, K.W.; Kann, H.E. Jr.

    1976-01-01

    The method of DNA alkaline elution was applied to a study of the formation and resealing of DNA single-strand breaks after irradiation of human fibroblasts with ultraviolet light (UV). The general features of the results were consistent with current concepts of DNA excision repair, in that breaks appeared rapidly after uv, and resealed slowly in normal fibroblasts, whereas breaks did not appear in those cells of patients with xeroderma pigmentosum (XP) that are known to have defects in DNA repair synthesis. The appearance of breaks required a short post-uv incubation, consistent with the expected action of an endonuclease. Cells of the variant form of XP characterized by normal DNA repair synthesis exhibited normal production of breaks after uv, but were slower than normal cells in resealing these breaks. This difference was enhanced by caffeine. A model is proposed to relate this finding with a previously described defect in post-replication repair in these XP variant cells. DNA crosslinking appears to cause an underestimate in the measurement of DNA breakage after uv

  9. Role of radiation chemical and enzymatic processes on single-strand breaks at short times after irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Sapora, O; Loverock, P S; Fielden, E M [Institute of Cancer Research, Sutton (UK). Surrey Branch

    1976-10-01

    A rapid mixing lysis technique has been used to study the effects of irradiation at different temperatures on two strains of E.coli K12, one lacking in the polymerase I activity (W3110), and the other carrying a ligase temperature-sensitive mutation (DY179), which had full ligase activity at 30/sup 0/C and none at 46/sup 0/C. The results provided direct evidence for the absence of any ligase-dependent repair of SSB at short times. The addition of 5 x 10/sup -3/M cysteine to heat-treated W3110 cells before irradiation in anoxic conditions practically removed the increase in yield of SSB per single strand genome shown by the heat-treated cells; the response was very close to that of normal cells in anoxia. The important contribution of sulphydryl compounds to the anoxic radio-biological response is thereby demonstrated. The basic difference in damage obtained by irradiation under oxic or anoxic conditions is due not to preferential enzymic (ligase) repair but to differences in radiation chemical events.

  10. Effect of Fluoride Prophylactic Agents on the Mechanical Properties of Nickel-Titanium Wires: An in vitro Study

    Directory of Open Access Journals (Sweden)

    S R Harish Koushik

    2011-01-01

    Conclusion : The results suggest that using topical fluoride agents with NiTi wire could decrease the functional unloading mechanical properties of the wire and contribute to prolonged orthodontic treatment.

  11. Mechanical properties of a new thermoplastic polymer orthodontic archwire

    Energy Technology Data Exchange (ETDEWEB)

    Varela, Juan Carlos; Velo, Marcos [Grupo de investigación en Ortodoncia, Facultad de Odontología, Universidad Santiago de Compostela, Santiago de Compostela (Spain); Espinar, Eduardo; Llamas, Jose Maria [Grupo de investigación en Ortodoncia, Facultad de Odontología, Universidad de Sevilla (Spain); Rúperez, Elisa; Manero, Jose Maria [Dept. C. Materiales e Ing. Metalúrgica, Universitat Politècnica de Catalunya, Centre de Recerca Nanoenginyeria, Member of Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN (Spain); Javier Gil, F., E-mail: francesc.xavier.gil@upc.edu [Dept. C. Materiales e Ing. Metalúrgica, Universitat Politècnica de Catalunya, Centre de Recerca Nanoenginyeria, Member of Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, CIBER-BBN (Spain)

    2014-09-01

    A new thermoplastic polymer for orthodontic applications was obtained and extruded into wires with round and rectangular cross sections. We evaluated the potential of new aesthetic archwire: tensile, three point bending, friction and stress relaxation behaviour, and formability characteristics were assessed. Stresses delivered were generally slightly lower than typical beta-titanium and nickel-titanium archwires. The polymer wire has good instantaneous mechanical properties; tensile stress decayed about 2% over 2 h depending on the initial stress relaxation for up to 120 h. High formability allowed shape bending similar to that associated with stainless steel wires. The friction coefficients were lower than the metallic conventional archwires improving the slipping with the brackets. This new polymer could be a good candidate for aesthetic orthodontic archwires. - Highlights: • A new thermoplastic polymer for orthodontic applications was obtained. • This polymer could be a good candidate for aesthetic orthodontic archwires. • The polymer has good mechanical properties as orthodontic wire coating. • The friction coefficients were lower than the metallic archwires improving the slipping with the brackets. • High formability allowed shape bending similar to that associated with stainless steel wires.

  12. Mechanical properties of a new thermoplastic polymer orthodontic archwire

    International Nuclear Information System (INIS)

    Varela, Juan Carlos; Velo, Marcos; Espinar, Eduardo; Llamas, Jose Maria; Rúperez, Elisa; Manero, Jose Maria; Javier Gil, F.

    2014-01-01

    A new thermoplastic polymer for orthodontic applications was obtained and extruded into wires with round and rectangular cross sections. We evaluated the potential of new aesthetic archwire: tensile, three point bending, friction and stress relaxation behaviour, and formability characteristics were assessed. Stresses delivered were generally slightly lower than typical beta-titanium and nickel-titanium archwires. The polymer wire has good instantaneous mechanical properties; tensile stress decayed about 2% over 2 h depending on the initial stress relaxation for up to 120 h. High formability allowed shape bending similar to that associated with stainless steel wires. The friction coefficients were lower than the metallic conventional archwires improving the slipping with the brackets. This new polymer could be a good candidate for aesthetic orthodontic archwires. - Highlights: • A new thermoplastic polymer for orthodontic applications was obtained. • This polymer could be a good candidate for aesthetic orthodontic archwires. • The polymer has good mechanical properties as orthodontic wire coating. • The friction coefficients were lower than the metallic archwires improving the slipping with the brackets. • High formability allowed shape bending similar to that associated with stainless steel wires

  13. Fibrotic encapsulation of orthodontic appliance in palate.

    Science.gov (United States)

    Muthu, Jananni; Muthanandam, Sivaramakrishnan; Umapathy, Gubernath; Kannan, Anitha Logaranjani

    2017-01-01

    Iatrogenic trauma though not serious is very common in dental practice. Orthodontic treatment can inflict such injuries as they are prolonged over a long period of time. Ill-fabricated orthodontic appliances, such as wires and brackets, or the patients' habits such as application of constant pressure over the appliance can traumatize the adjacent oral soft tissues. In rare cases, these appliances can get embedded into the mucosa and gingival tissues. This case report describes one such case of iatrogenic trauma to the palatal mucosa due to entrapment of a tongue spike appliance and its surgical management.

  14. Three-dimensional deformation of orthodontic brackets

    Science.gov (United States)

    Melenka, Garrett W; Nobes, David S; Major, Paul W

    2013-01-01

    Braces are used by orthodontists to correct the misalignment of teeth in the mouth. Archwire rotation is a particular procedure used to correct tooth inclination. Wire rotation can result in deformation to the orthodontic brackets, and an orthodontic torque simulator has been designed to examine this wire–bracket interaction. An optical technique has been employed to measure the deformation due to size and geometric constraints of the orthodontic brackets. Images of orthodontic brackets are collected using a stereo microscope and two charge-coupled device cameras, and deformation of orthodontic brackets is measured using a three-dimensional digital image correlation technique. The three-dimensional deformation of orthodontic brackets will be evaluated. The repeatability of the three-dimensional digital image correlation measurement method was evaluated by performing 30 archwire rotation tests using the same bracket and archwire. Finally, five Damon 3MX and five In-Ovation R self-ligating brackets will be compared using this technique to demonstrate the effect of archwire rotation on bracket design. PMID:23762201

  15. Trends in contemporary orthodontic research publications: Evaluation of three major orthodontic journals

    Directory of Open Access Journals (Sweden)

    M P Kumar Prasanna

    2017-01-01

    Full Text Available Background: In this study, we aimed to evaluate the inclination of orthodontic research published in original articles in three of the most popular and recognized orthodontic journals with high impact factor; American Journal of Orthodontics and Dentofacial Orthopedics (AJODO, the Angle Orthodontist (AO, and European Journal of Orthodontics (EJO published in a 5-year duration time frame (2010–2014. Materials and Methods: Online search with supplementary hand searching was undertaken for original research articles in these three orthodontic journals from 2010 to 2014. Classification of data was completed autonomously by manual and direct appraisal of the manuscript of each journal. Results and Conclusion: Three thousand one hundred and forty articles published in AJODO, AO, and EJO in the particular time period were selected, out of which 1783 original research study articles (56.78% were appraised for classification in their various respective knowledge domains. AO (82.73% and EJO (76.99% were relatively more committed toward publication of research work in comparison to AJODO (34.55%. Research articles based on perception and questionnaires (12%, temporary anchorage devices (11%, cone-beam computed tomography (8%, invisible orthodontics (lingual orthodontics, clear aligners, esthetic brackets, and wires (5%, and airway examination (5% were the five most recurrently selected subjects of interest (41% in these three journals during the observation period. The total number of research articles published in the three journals reduced statistically significantly in 2014 when compared to 2010.

  16. TERRA and hnRNPA1 orchestrate an RPA-to-POT1 switch on telomeric single-stranded DNA.

    Science.gov (United States)

    Flynn, Rachel Litman; Centore, Richard C; O'Sullivan, Roderick J; Rai, Rekha; Tse, Alice; Songyang, Zhou; Chang, Sandy; Karlseder, Jan; Zou, Lee

    2011-03-24

    Maintenance of telomeres requires both DNA replication and telomere 'capping' by shelterin. These two processes use two single-stranded DNA (ssDNA)-binding proteins, replication protein A (RPA) and protection of telomeres 1 (POT1). Although RPA and POT1 each have a critical role at telomeres, how they function in concert is not clear. POT1 ablation leads to activation of the ataxia telangiectasia and Rad3-related (ATR) checkpoint kinase at telomeres, suggesting that POT1 antagonizes RPA binding to telomeric ssDNA. Unexpectedly, we found that purified POT1 and its functional partner TPP1 are unable to prevent RPA binding to telomeric ssDNA efficiently. In cell extracts, we identified a novel activity that specifically displaces RPA, but not POT1, from telomeric ssDNA. Using purified protein, here we show that the heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) recapitulates the RPA displacing activity. The RPA displacing activity is inhibited by the telomeric repeat-containing RNA (TERRA) in early S phase, but is then unleashed in late S phase when TERRA levels decline at telomeres. Interestingly, TERRA also promotes POT1 binding to telomeric ssDNA by removing hnRNPA1, suggesting that the re-accumulation of TERRA after S phase helps to complete the RPA-to-POT1 switch on telomeric ssDNA. Together, our data suggest that hnRNPA1, TERRA and POT1 act in concert to displace RPA from telomeric ssDNA after DNA replication, and promote telomere capping to preserve genomic integrity.

  17. Replication protein A (RPA) hampers the processive action of APOBEC3G cytosine deaminase on single-stranded DNA.

    Science.gov (United States)

    Lada, Artem G; Waisertreiger, Irina S-R; Grabow, Corinn E; Prakash, Aishwarya; Borgstahl, Gloria E O; Rogozin, Igor B; Pavlov, Youri I

    2011-01-01

    Editing deaminases have a pivotal role in cellular physiology. A notable member of this superfamily, APOBEC3G (A3G), restricts retroviruses, and Activation Induced Deaminase (AID) generates antibody diversity by localized deamination of cytosines in DNA. Unconstrained deaminase activity can cause genome-wide mutagenesis and cancer. The mechanisms that protect the genomic DNA from the undesired action of deaminases are unknown. Using the in vitro deamination assays and expression of A3G in yeast, we show that replication protein A (RPA), the eukaryotic single-stranded DNA (ssDNA) binding protein, severely inhibits the deamination activity and processivity of A3G. We found that mutations induced by A3G in the yeast genomic reporter are changes of a single nucleotide. This is unexpected because of the known property of A3G to catalyze multiple deaminations upon one substrate encounter event in vitro. The addition of recombinant RPA to the oligonucleotide deamination assay severely inhibited A3G activity. Additionally, we reveal the inverse correlation between RPA concentration and the number of deaminations induced by A3G in vitro on long ssDNA regions. This resembles the "hit and run" single base substitution events observed in yeast. Our data suggest that RPA is a plausible antimutator factor limiting the activity and processivity of editing deaminases in the model yeast system. Because of the similar antagonism of yeast RPA and human RPA with A3G in vitro, we propose that RPA plays a role in the protection of the human genome cell from A3G and other deaminases when they are inadvertently diverged from their natural targets. We propose a model where RPA serves as one of the guardians of the genome that protects ssDNA from the destructive processive activity of deaminases by non-specific steric hindrance.

  18. Thermodynamic characterization of binding Oxytricha nova single strand telomere DNA with the alpha protein N-terminal domain.

    Science.gov (United States)

    Buczek, Pawel; Horvath, Martin P

    2006-06-23

    The Oxytricha nova telemere binding protein alpha subunit binds single strand DNA and participates in a nucleoprotein complex that protects the very ends of chromosomes. To understand how the N-terminal, DNA binding domain of alpha interacts with DNA we measured the stoichiometry, enthalpy (DeltaH), entropy (DeltaS), and dissociation constant (K(D-DNA)) for binding telomere DNA fragments at different temperatures and salt concentrations using native gel electrophoresis and isothermal titration calorimetry (ITC). About 85% of the total free energy of binding corresponded with non-electrostatic interactions for all DNAs. Telomere DNA fragments d(T(2)G(4)), d(T(4)G(4)), d(G(3)T(4)G(4)), and d(G(4)T(4)G(4)) each formed monovalent protein complexes. In the case of d(T(4)G(4)T(4)G(4)), which has two tandemly repeated d(TTTTTGGGG) telomere motifs, two binding sites were observed. The high-affinity "A site" has a dissociation constant, K(D-DNA(A)) = 13(+/-4) nM, while the low-affinity "B site" is characterized by K(D-DNA(B)) = 5600(+/-600) nM at 25 degrees C. Nucleotide substitution variants verified that the A site corresponds principally with the 3'-terminal portion of d(T(4)G(4)T(4)G(4)). The relative contributions of entropy (DeltaS) and enthalpy (DeltaH) for binding reactions were DNA length-dependent as was heat capacity (DeltaCp). These trends with respect to DNA length likely reflect structural transitions in the DNA molecule that are coupled with DNA-protein association. Results presented here are important for understanding early intermediates and subsequent stages in the assembly of the full telomere nucleoprotein complex and how binding events can prepare the telomere DNA for extension by telomerase, a critical event in telomere biology.

  19. All-atom molecular dynamics simulations of spin labelled double and single-strand DNA for EPR studies.

    Science.gov (United States)

    Prior, C; Danilāne, L; Oganesyan, V S

    2018-05-16

    We report the first application of fully atomistic molecular dynamics (MD) simulations to the prediction of electron paramagnetic resonance (EPR) spectra of spin labelled DNA. Models for two structurally different DNA spin probes with either the rigid or flexible position of the nitroxide group in the base pair, employed in experimental studies previously, have been developed. By the application of the combined MD-EPR simulation methodology we aimed at the following. Firstly, to provide a test bed against a sensitive spectroscopic technique for the recently developed improved version of the parmbsc1 force field for MD modelling of DNA. The predicted EPR spectra show good agreement with the experimental ones available from the literature, thus confirming the accuracy of the currently employed DNA force fields. Secondly, to provide a quantitative interpretation of the motional contributions into the dynamics of spin probes in both duplex and single-strand DNA fragments and to analyse their perturbing effects on the local DNA structure. Finally, a combination of MD and EPR allowed us to test the validity of the application of the Model-Free (M-F) approach coupled with the partial averaging of magnetic tensors to the simulation of EPR spectra of DNA systems by comparing the resultant EPR spectra with those simulated directly from MD trajectories. The advantage of the M-F based EPR simulation approach over the direct propagation techniques is that it requires motional and order parameters that can be calculated from shorter MD trajectories. The reported MD-EPR methodology is transferable to the prediction and interpretation of EPR spectra of higher order DNA structures with novel types of spin labels.

  20. Evidence of pervasive biologically functional secondary structures within the genomes of eukaryotic single-stranded DNA viruses.

    Science.gov (United States)

    Muhire, Brejnev Muhizi; Golden, Michael; Murrell, Ben; Lefeuvre, Pierre; Lett, Jean-Michel; Gray, Alistair; Poon, Art Y F; Ngandu, Nobubelo Kwanele; Semegni, Yves; Tanov, Emil Pavlov; Monjane, Adérito Luis; Harkins, Gordon William; Varsani, Arvind; Shepherd, Dionne Natalie; Martin, Darren Patrick

    2014-02-01

    Single-stranded DNA (ssDNA) viruses have genomes that are potentially capable of forming complex secondary structures through Watson-Crick base pairing between their constituent nucleotides. A few of the structural elements formed by such base pairings are, in fact, known to have important functions during the replication of many ssDNA viruses. Unknown, however, are (i) whether numerous additional ssDNA virus genomic structural elements predicted to exist by computational DNA folding methods actually exist and (ii) whether those structures that do exist have any biological relevance. We therefore computationally inferred lists of the most evolutionarily conserved structures within a diverse selection of animal- and plant-infecting ssDNA viruses drawn from the families Circoviridae, Anelloviridae, Parvoviridae, Nanoviridae, and Geminiviridae and analyzed these for evidence of natural selection favoring the maintenance of these structures. While we find evidence that is consistent with purifying selection being stronger at nucleotide sites that are predicted to be base paired than at sites predicted to be unpaired, we also find strong associations between sites that are predicted to pair with one another and site pairs that are apparently coevolving in a complementary fashion. Collectively, these results indicate that natural selection actively preserves much of the pervasive secondary structure that is evident within eukaryote-infecting ssDNA virus genomes and, therefore, that much of this structure is biologically functional. Lastly, we provide examples of various highly conserved but completely uncharacterized structural elements that likely have important functions within some of the ssDNA virus genomes analyzed here.

  1. DNA translocation by human uracil DNA glycosylase: the case of single-stranded DNA and clustered uracils.

    Science.gov (United States)

    Schonhoft, Joseph D; Stivers, James T

    2013-04-16

    Human uracil DNA glycosylase (hUNG) plays a central role in DNA repair and programmed mutagenesis of Ig genes, requiring it to act on sparsely or densely spaced uracil bases located in a variety of contexts, including U/A and U/G base pairs, and potentially uracils within single-stranded DNA (ssDNA). An interesting question is whether the facilitated search mode of hUNG, which includes both DNA sliding and hopping, changes in these different contexts. Here we find that hUNG uses an enhanced local search mode when it acts on uracils in ssDNA, and also, in a context where uracils are densely clustered in duplex DNA. In the context of ssDNA, hUNG performs an enhanced local search by sliding with a mean sliding length larger than that of double-stranded DNA (dsDNA). In the context of duplex DNA, insertion of high-affinity abasic product sites between two uracil lesions serves to significantly extend the apparent sliding length on dsDNA from 4 to 20 bp and, in some cases, leads to directionally biased 3' → 5' sliding. The presence of intervening abasic product sites mimics the situation where hUNG acts iteratively on densely spaced uracils. The findings suggest that intervening product sites serve to increase the amount of time the enzyme remains associated with DNA as compared to nonspecific DNA, which in turn increases the likelihood of sliding as opposed to falling off the DNA. These findings illustrate how the search mechanism of hUNG is not predetermined but, instead, depends on the context in which the uracils are located.

  2. Replication protein A (RPA hampers the processive action of APOBEC3G cytosine deaminase on single-stranded DNA.

    Directory of Open Access Journals (Sweden)

    Artem G Lada

    Full Text Available Editing deaminases have a pivotal role in cellular physiology. A notable member of this superfamily, APOBEC3G (A3G, restricts retroviruses, and Activation Induced Deaminase (AID generates antibody diversity by localized deamination of cytosines in DNA. Unconstrained deaminase activity can cause genome-wide mutagenesis and cancer. The mechanisms that protect the genomic DNA from the undesired action of deaminases are unknown. Using the in vitro deamination assays and expression of A3G in yeast, we show that replication protein A (RPA, the eukaryotic single-stranded DNA (ssDNA binding protein, severely inhibits the deamination activity and processivity of A3G.We found that mutations induced by A3G in the yeast genomic reporter are changes of a single nucleotide. This is unexpected because of the known property of A3G to catalyze multiple deaminations upon one substrate encounter event in vitro. The addition of recombinant RPA to the oligonucleotide deamination assay severely inhibited A3G activity. Additionally, we reveal the inverse correlation between RPA concentration and the number of deaminations induced by A3G in vitro on long ssDNA regions. This resembles the "hit and run" single base substitution events observed in yeast.Our data suggest that RPA is a plausible antimutator factor limiting the activity and processivity of editing deaminases in the model yeast system. Because of the similar antagonism of yeast RPA and human RPA with A3G in vitro, we propose that RPA plays a role in the protection of the human genome cell from A3G and other deaminases when they are inadvertently diverged from their natural targets. We propose a model where RPA serves as one of the guardians of the genome that protects ssDNA from the destructive processive activity of deaminases by non-specific steric hindrance.

  3. Analysis of the substrate recognition state of TDP-43 to single-stranded DNA using fluorescence correlation spectroscopy

    Directory of Open Access Journals (Sweden)

    Akira Kitamura

    2018-07-01

    Full Text Available Normal function and abnormal aggregation of transactivation response (TAR DNA/RNA-binding protein 43 kDa (TDP-43 are directly associated with the lethal genetic diseases: cystic fibrosis, amyotrophic lateral sclerosis (ALS, and frontotemporal lobar degeneration (FTLD. The binding of TDP-43 to single-stranded DNA (ssDNA or RNA is involved in transcriptional repression, regulation of RNA splicing, and RNA stabilization. Equilibrium dissociation constants (Kd of TDP-43 and ssDNA or RNA have been determined using various methods; however, methods that can measure Kd with high sensitivity in a short time using a small amount of TDP-43 in solution would be advantageous. Here, in order to determine the Kd of TDP-43 and fluorescence-labeled ssDNA as well as the binding stoichiometry, we use fluorescence correlation spectroscopy (FCS, which detects the slowed diffusion of molecular interactions in solution with single-molecule sensitivity, in addition to electrophoretic mobility shift assay (EMSA. Using tandem affinity chromatography of TDP-43 dually tagged with glutathione-S-transferase and poly-histidine tags, highly purified protein was obtained. FCS successfully detected specific interaction between purified TDP-43 and TG ssDNA repeats, with a Kd in the nanomolar range. The Kd of the TDP-43 mutant was not different from the wild type, although mutant oligomers, which did not bind ssDNA, were observed. Analysis of the fluorescence brightness per dimerized TDP-43/ssDNA complex was used to evaluate their binding stoichiometry. The results suggest that an assay combining FCS and EMSA can precisely analyze ssDNA recognition mechanisms, and that FCS may be applied for the rapid and quantitative determination of the interaction strength between TDP-43 and ssDNA or RNA. These methods will aid in the elucidation of the substrate recognition mechanism of ALS- and FTLD-associated variants of TDP-43.

  4. Viral recombination blurs taxonomic lines: examination of single-stranded DNA viruses in a wastewater treatment plant

    Directory of Open Access Journals (Sweden)

    Victoria M. Pearson

    2016-10-01

    Full Text Available Understanding the structure and dynamics of microbial communities, especially those of economic concern, is of paramount importance to maintaining healthy and efficient microbial communities at agricultural sites and large industrial cultures, including bioprocessors. Wastewater treatment plants are large bioprocessors which receive water from multiple sources, becoming reservoirs for the collection of many viral families that infect a broad range of hosts. To examine this complex collection of viruses, full-length genomes of circular ssDNA viruses were isolated from a wastewater treatment facility using a combination of sucrose-gradient size selection and rolling-circle amplification and sequenced on an Illumina MiSeq. Single-stranded DNA viruses are among the least understood groups of microbial pathogens due to genomic biases and culturing difficulties, particularly compared to the larger, more often studied dsDNA viruses. However, the group contains several notable well-studied examples, including agricultural pathogens which infect both livestock and crops (Circoviridae and Geminiviridae, and model organisms for genetics and evolution studies (Microviridae. Examination of the collected viral DNA provided evidence for 83 unique genotypic groupings, which were genetically dissimilar to known viral types and exhibited broad diversity within the community. Furthermore, although these genomes express similarities to known viral families, such as Circoviridae, Geminiviridae, and Microviridae, many are so divergent that they may represent new taxonomic groups. This study demonstrated the efficacy of the protocol for separating bacteria and large viruses from the sought after ssDNA viruses and the ability to use this protocol to obtain an in-depth analysis of the diversity within this group.

  5. UPregulated single-stranded DNA-binding protein 1 induces cell chemoresistance to cisplatin in lung cancer cell lines.

    Science.gov (United States)

    Zhao, Xiang; He, Rong; Liu, Yu; Wu, Yongkai; Kang, Leitao

    2017-07-01

    Cisplatin and its analogues are widely used as anti-tumor drugs in lung cancer but many cisplatin-resistant lung cancer cases have been identified in recent years. Single-stranded DNA-binding protein 1 (SSDBP1) can effectively induce H69 cell resistance to cisplatin in our previous identification; thus, it is necessary to explore the mechanism underlying the effects of SSDBP1-induced resistance to cisplatin. First, SSDBP1-overexpressed or silent cell line was constructed and used to analyze the effects of SSDBP1 on chemoresistance of lung cancer cells to cisplatin. SSDBP1 expression was assayed by real-time PCR and Western blot. Next, the effects of SSDBP1 on cisplatin sensitivity, proliferation, and apoptosis of lung cancer cell lines were assayed by MTT and flow cytometry, respectively; ABC transporters, apoptosis-related genes, and cell cycle-related genes by real-time PCR, and DNA wound repair by comet assay. Low expression of SSDBP1 was observed in H69 cells, while increased expression in cisplatin-resistant H69 cells. Upregulated expression of SSDBP1 in H69AR cells was identified to promote proliferation and cisplatin resistance and inhibit apoptosis, while downregulation of SSDBP1 to inhibit cisplatin resistance and proliferation and promoted apoptosis. Moreover, SSDBP1 promoted the expression of P2gp, MRP1, Cyclin D1, and CDK4 and inhibited the expression of caspase 3 and caspase 9. Furthermore, SSDBP1 promoted the DNA wound repair. These results indicated that SSDBP1 may induce cell chemoresistance of cisplatin through promoting DNA repair, resistance-related gene expression, cell proliferation, and inhibiting apoptosis.

  6. Sequence-based separation of single-stranded DNA using nucleotides in capillary electrophoresis: focus on phosphate.

    Science.gov (United States)

    Zhang, Xueru; McGown, Linda B

    2013-06-01

    DNA analysis has widespread applicability in biology, medicine, biotechnology, and forensics. DNA separation by length is readily achieved using sieving gels in electrophoresis. Separation by sequence is less simple, generally requiring adequate differences in native or induced conformation or differences in thermal or chemical stability of the strands that are hybridized prior to measurement. We previously demonstrated separation of four single-stranded DNA 76-mers that differ by only a few A-G substitutions based solely on sequence using guanosine-5'-monophosphate (GMP) in the running buffer. We attributed separation to the unique self-assembly of GMP to form higher order structures. Here, we examine an expanded set of 76-mers designed to probe the mechanism of the separation and effects of experimental conditions. We were surprised to find that other ribonucleotides achieved the similar separation to GMP, and that some separation was achieved using sodium phosphate instead of GMP. Potassium phosphate achieved almost as good separations as the ribonucleotides. This suggests that the separation medium provides a physicochemical environment for the DNA that effects strand migration in a sequence-selective manner. Further investigation is needed to determine whether the mechanism involves specific interactions between the phosphates and the DNA strands or is a result of other properties of the separation medium. Phosphate generally has been avoided in DNA separations by capillary gel electrophoresis because its high ionic strength exacerbates Joule heating. Our results suggest that phosphate compounds should be examined for separation of DNA based on sequence. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Nucleotide fluctuation of radiation-resistant Halobacterium sp. NRC-1 single-stranded DNA-binding protein (RPA) genes

    Science.gov (United States)

    Holden, Todd; Tremberger, G., Jr.; Cheung, E.; Subramaniam, R.; Gadura, N.; Schneider, P.; Sullivan, R.; Flamholz, A.; Lieberman, D.; Cheung, T. D.

    2009-08-01

    The Single-Stranded DNA-Binding Protein (RPA) Genes in gamma ray radiation-resistant halophilic archaeon Halobacterium sp. NRC-1 were analyzed in terms of their nucleotide fluctuations. In an ATCG sequence, each base was assigned a number equal to its atomic number. The resulting numerical sequence was the basis of the statistical analysis in this study. Fractal analysis using the Higuchi method gave fractal dimensions of 2.04 and 2.06 for the gene sequences VNG2160 and VNG2162, respectively. The 16S rRNA sequence has a fractal dimension of 1.99. The di-nucleotide Shannon entropy values were found to be negatively correlated with the observed fractal dimensions (R2~ 0.992, N=3). Inclusion of Deinococcus radiodurans Rad-A in the regression analysis decreases the R2 slightly to 0.98 (N=4). A third VNG2163 RPA gene of unknown function but with upregulation activity under irradiation was found to have a fractal dimension of 2.05 and a Shannon entropy of 3.77 bits. The above results are similar to those found in bacterial Deinococcus radiodurans and suggest that their high radiation resistance property would have favored selection of CG di-nucleotide pairs. The two transcription factors TbpD (VNG7114) and TfbA (VNG 2184) were also studied. Using VNG7114, VNG2184, and VNG2163; the regression analysis of fractal dimension versus Shannon entropy shows that R2 ~ 0.997 for N =3. The VNG2163 unknown function may be related to the pathways with transcriptions closely regulated to sequences VNG7114 and VNG2184.

  8. Ultra-fast repair of single-strand breaks in DNA of. gamma. -irradiated Chinese hamster cells

    Energy Technology Data Exchange (ETDEWEB)

    Leontjeva, G A; Mantzighin, Yu A; Gaziev, A I [AN SSSR, Pushchino-na-Oke. Inst. Biologicheskoj Fiziki

    1976-12-01

    Studies of the effect of thermal treatment of Chinese hamster cells on sedimentation of DNA in the alkaline sucrose gradient showed that heating the cells to 68/sup 0/C for 15 min caused the same degradation as ..gamma..-irradiation with 5 to 7 krad at 37/sup 0/C. The inhibition of cellular repair enzymes by heating was therefore unacceptable. The process of ultra-fast repair is essentially determined by the DNA-ligase reaction, which is activated in the presence of Mg ions, and inhibited in mammalian cells in the presence of EDTA and pyrophosphate. Sedimentation profiles were therefore measured for the DNA of Chinese hamster cells ..gamma..-irradiated (5 krad) at 0/sup 0/C or 22/sup 0/C in the presence of Mg/sup + +/, or EDTA and pyrophosphate, and the results demonstrated ultra-fast repair only at 20 to 37/sup 0/C, in contrast to bacteria. A study was made of the temperature dependence of the activity of the DNA ligases isolated from E.coli and rabbit bone marrow. The NAD-dependent bacterial DNA ligase was active at temperatures from 0 to 40/sup 0/C, whereas ATP-dependent DNA ligase of mammals only showed activity in the range 15 to 40/sup 0/C. The differing temperature dependences of ultra-fast repair in bacterial and mammalian cells are in agreement with the temperature dependences of the activities of isolated enzymes, and the results suggest that the process of ultra-fast repair of single-strand breaks of DNA takes place in both bacterial and mammalian cells.

  9. Dental Extrusion with Orthodontic Miniscrew Anchorage: A Case Report Describing a Modified Method

    OpenAIRE

    Horliana, Ricardo Fidos; Horliana, Anna Carolina Ratto Tempestini; Wuo, Alexandre do Vale; Perez, Fl?vio Eduardo Guillin; Abr?o, Jorge

    2015-01-01

    In recent years, the skeletal anchorage through miniscrews has expanded the treatment options in orthodontics (Yamaguchi et al., 2012). We hereby present a modified method for tooth extrusion for cases where crown-lengthening surgery is contraindicated for aesthetic reasons. This modified method uses three orthodontic appliances: a mini-implant, an orthodontic wire, and a bracket. The aim of this case report was to increase the length of the clinical crown of a fractured tooth (tooth 23) by m...

  10. Construction of a microfluidic chip, using dried-down reagents, for LATE-PCR amplification and detection of single-stranded DNA.

    Science.gov (United States)

    Jia, Yanwei; Mak, Pui-In; Massey, Conner; Martins, Rui P; Wangh, Lawrence J

    2013-12-07

    LATE-PCR is an advanced form of non-symmetric PCR that efficiently generates single-stranded DNA which can readily be characterized at the end of amplification by hybridization to low-temperature fluorescent probes. We demonstrate here for the first time that monoplex and duplex LATE-PCR amplification and probe target hybridization can be carried out in double layered PDMS microfluidics chips containing dried reagents. Addition of a set of reagents during dry down overcomes the common problem of single-stranded oligonucleotide binding to PDMS. These proof-of-principle results open the way to construction of inexpensive point-of-care devices that take full advantage of the analytical power of assays built using LATE-PCR and low-temperature probes.

  11. Hot topic: Bovine milk samples yielding negative or nonspecific results in bacterial culturing--the possible role of PCR-single strand conformation polymorphism in mastitis diagnosis.

    Science.gov (United States)

    Schwaiger, K; Wimmer, M; Huber-Schlenstedt, R; Fehlings, K; Hölzel, C S; Bauer, J

    2012-01-01

    A large proportion of mastitis milk samples yield negative or nonspecific results (i.e., no mastitis pathogen can be identified) in bacterial culturing. Therefore, the culture-independent PCR-single strand conformation polymorphism method was applied to the investigation of bovine mastitis milk samples. In addition to the known mastitis pathogens, the method was suitable for the detection of fastidious bacteria such as Mycoplasma spp., which are often missed by conventional culturing methods. The detection of Helcococcus ovis in 4 samples might indicate an involvement of this species in pathogenesis of bovine mastitis. In conclusion, PCR-single-strand conformation polymorphism is a promising tool for gaining new insights into the bacteriological etiology of mastitis. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  12. Genetic effects and reparation of single-stranded DNA breaks in Arabidopsis thaliana populations growing in the vicinity of the Chernobyl Nuclear Power Station

    International Nuclear Information System (INIS)

    Abramov, V.I.; Sergeeva, S.A.; Ptitsyna, S.N.; Semov, A.B.; Shevchenko, V.A.

    1992-01-01

    The genetic effects and efficiency of repair of single-stranded DNA breaks in natural populations of Arabidopsis growing within a thirty-kilometer zone of the Chernobyl Nuclear Power Station were studied. A direct relationship was found between the level of radioactive contamination and the frequency of embryonal lethal mutations in the Arabidopsis populations studied. A decrease in the efficiency of reparation of single-stranded DNA breaks was found in Arabidopsis plants growing in the contaminated sites. The level of efficiency of DNA reparation was dependent on the duration for which the Arabidopsis population had been growing in the contaminated sites and on the degree of radioactive contamination of the sites. 9 refs., 4 tabs

  13. DNA polymerase I-mediated repair of 365 nm-induced single-strand breaks in the DNA of Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Ley, R D; Sedita, B A; Boye, E [Argonne National Lab., Ill. (USA)

    1978-03-01

    Irradiation of closed circular phage lambda DNA in vivo at 365 nm results in the induction of single-strand breaks and alkali-labile lesions at rates of 1.1 x 10/sup -14/ and 0.2 x 10/sup -14//dalton/J/m/sup 2/, respectively. The sum of the induction rates is similar to the rate of induction of single-strand breaks plus alkali-labile lesions (1 x 10/sup -14//dalton/J/m/sup 2/) observed in the E. coli genome. Postirradiation incubation of wild-type cells in buffer results in rapid repair of the breaks (up to 80% repaired in 10 min). No repair was observed in a DNA polymerase I-deficient mutant of E.coli.

  14. Impaction of Maxillary Central Incisors: Surgical and Orthodontic Treatment--Case Report.

    Science.gov (United States)

    Knop, Luegya Amorim Henriques; Shintcovsk, Ricardo Lima; Gandini, Luiz Gonzaga; Parsekian, Lidia; Pinto, Ary Santos

    2015-01-01

    The aim of this paper is to report two clinical cases, which was performed with surgical exposure and orthodontic traction of a maxillary central incisors. Light forces were used during the orthodontic treatment applied on rigid wires as anchorage. It was noted that teeth presented adequate clinical crown height and gingival contours.

  15. Rolling replication of UV-irradiated duplex DNA in the phi X174 replicative-form----single-strand replication system in vitro

    International Nuclear Information System (INIS)

    Shavitt, O.; Livneh, Z.

    1989-01-01

    Cloning of the phi X174 viral origin of replication into phage M13mp8 produced an M13-phi X174 chimera, the DNA of which directed efficient replicative-form----single-strand rolling replication in vitro. This replication assay was performed with purified phi X174-encoded gene A protein, Escherichia coli rep helicase, single-stranded DNA-binding protein, and DNA polymerase III holoenzyme. The nicking of replicative-form I (RFI) DNA by gene A protein was essentially unaffected by the presence of UV lesions in the DNA. However, unwinding of UV-irradiated DNA by the rep helicase was inhibited twofold as compared with unwinding of the unirradiated substrate. UV irradiation of the substrate DNA caused a strong inhibition in its ability to direct DNA synthesis. However, even DNA preparations that contained as many as 10 photodimers per molecule still supported the synthesis of progeny full-length single-stranded DNA. The appearance of full-length radiolabeled products implied at least two full rounds of replication, since the first round released the unlabeled plus viral strand of the duplex DNA. Pretreatment of the UV-irradiated DNA substrate with purified pyrimidine dimer endonuclease from Micrococcus luteus, which converted photodimer-containing supercoiled RFI DNA into relaxed, nicked RFII DNA and thus prevented its replication, reduced DNA synthesis by 70%. Analysis of radiolabeled replication products by agarose gel electrophoresis followed by autoradiography revealed that this decrease was due to a reduction in the synthesis of progeny full-length single-stranded DNA. This implies that 70 to 80% of the full-length DNA products produced in this system were synthesized on molecules that carried photodimers

  16. Genotyping of human parvovirus B19 in clinical samples from Brazil and Paraguay using heteroduplex mobility assay, single-stranded conformation polymorphism and nucleotide sequencing

    Directory of Open Access Journals (Sweden)

    Marcos César Lima de Mendonça

    2011-06-01

    Full Text Available Heteroduplex mobility assay, single-stranded conformation polymorphism and nucleotide sequencing were utilised to genotype human parvovirus B19 samples from Brazil and Paraguay. Ninety-seven serum samples were collected from individuals presenting with abortion or erythema infectiosum, arthropathies, severe anaemia and transient aplastic crisis; two additional skin samples were collected by biopsy. After the procedure, all clinical samples were classified as genotype 1.

  17. SALP, a new single-stranded DNA library preparation method especially useful for the high-throughput characterization of chromatin openness states.

    Science.gov (United States)

    Wu, Jian; Dai, Wei; Wu, Lin; Wang, Jinke

    2018-02-13

    Next-generation sequencing (NGS) is fundamental to the current biological and biomedical research. Construction of sequencing library is a key step of NGS. Therefore, various library construction methods have been explored. However, the current methods are still limited by some shortcomings. This study developed a new NGS library construction method, Single strand Adaptor Library Preparation (SALP), by using a novel single strand adaptor (SSA). SSA is a double-stranded oligonucleotide with a 3' overhang of 3 random nucleotides, which can be efficiently ligated to the 3' end of single strand DNA by T4 DNA ligase. SALP can be started with any denatured DNA fragments such as those sheared by Tn5 tagmentation, enzyme digestion and sonication. When started with Tn5-tagmented chromatin, SALP can overcome a key limitation of ATAC-seq and become a high-throughput NGS library construction method, SALP-seq, which can be used to comparatively characterize the chromatin openness state of multiple cells unbiasly. In this way, this study successfully characterized the comparative chromatin openness states of four different cell lines, including GM12878, HepG2, HeLa and 293T, with SALP-seq. Similarly, this study also successfully characterized the chromatin openness states of HepG2 cells with SALP-seq by using 10 5 to 500 cells. This study developed a new NGS library construction method, SALP, by using a novel kind of single strand adaptor (SSA), which should has wide applications in the future due to its unique performance.

  18. Coupled aggregation of mitochondrial single-strand DNA-binding protein tagged with Eos fluorescent protein visualizes synchronized activity of mitochondrial nucleoids

    Czech Academy of Sciences Publication Activity Database

    Olejár, Tomáš; Pajuelo-Reguera, David; Alán, Lukáš; Dlasková, Andrea; Ježek, Petr

    2015-01-01

    Roč. 12, č. 4 (2015), s. 5185-5190 ISSN 1791-2997 R&D Projects: GA ČR(CZ) GAP302/10/0346; GA MŠk(CZ) EE2.3.30.0025 Institutional support: RVO:67985823 Keywords : mitochondrial nucleoid * single-stranded DNA-binding protein * photoconvertible fluorescent protein Eos Subject RIV: EA - Cell Biology Impact factor: 1.559, year: 2015

  19. The survival and repair of DNA single-strand breaks in gamma-irradiated Escherichia coli adapted to methyl methane sulfonate

    International Nuclear Information System (INIS)

    Zhestyanikov, V.D.; Savel'eva, G.E.

    1992-01-01

    The survival and repair of single-strand breaks of DNA in gamma-irradiated E.coli adapted to methyl methane sulfonate (MMS) (20 mkg/ml during 3 hours) have been investigated. It is shown that the survival of adapted bacteria of radioresistant strains B/r, H/r30, AB1157 and W3110 pol + increases with DMF (dose modification factor) ranging within 1.4-1.8 and in radiosensitive strains B s-1 , AB1157 recA13 and AB1157 lexA3 with DMF ranging within 1.3-1.4, and does not change in strains with mutation in poLA gene P3478 poLA1 and 016 res-3. The increase in radioresistance during the adaptation to MMS correlates with the acceleration of repair of gamma-ray-induced single-strand breaks in the radioresistant strains B/r and W3110 pol + and with the appearance of the ability to repair some part of DNA single-strand breaks in the mutant B s-1

  20. Reduction of spontaneous somatic mutation frequency by a low-dose X irradiation of Drosophila larvae and possible involvement of DNA single-strand damage repair.

    Science.gov (United States)

    Koana, Takao; Takahashi, Takashi; Tsujimura, Hidenobu

    2012-03-01

    The third instar larvae of Drosophila were irradiated with X rays, and the somatic mutation frequency in their wings was measured after their eclosion. In the flies with normal DNA repair and apoptosis functions, 0.2 Gy irradiation at 0.05 Gy/min reduced the frequency of the so-called small spot (mutant cell clone with reduced reproductive activity) compared with that in the sham-irradiated flies. When apoptosis was suppressed using the baculovirus p35 gene, the small spot frequency increased four times in the sham-irradiated control group, but the reduction by the 0.2-Gy irradiation was still evident. In a non-homologous end joining-deficient mutant, the small spot frequency was also reduced by 0.2 Gy radiation. In a mutant deficient in single-strand break repair, no reduction in the small spot frequency by 0.2 Gy radiation was observed, and the small spot frequency increased with the radiation dose. Large spot (mutant cell clone with normal reproductive activity) frequency was not affected by suppression of apoptosis and increased monotonically with radiation dose in wild-type larvae and in mutants for single- or double-strand break repair. It is hypothesized that some of the small spots resulted from single-strand damage and, in wild-type larvae, 0.2 Gy radiation activated the normal single-strand break repair gene, which reduced the background somatic mutation frequency.

  1. The validity of sedimentation data from high molecular weight DNA and the effects of additives on radiation-induced single-strand breakage

    International Nuclear Information System (INIS)

    Dugle, D.L.

    1979-10-01

    The optimization of many of the factors governing reproducible sedimentation behaviour of high molecular weight single-strand DNA in a particular alkaline sucrose density gradient system is described. A range of angular momenta is defined for which a constant strand breakage efficiency is required, despite a rotor speed effect which increases the measured molecular weights at decreasing rotor speeds for larger DNA molecules. The possibility is discussed that the bimodal control DNA profiles obtained after sedimentation at 11 500 rev/min (12 400 g) or less represent structural subunits of the chromatid. The random induction of single-strand DNA breaks by ionizing radiation is demonstrated by the computer-derived fits to the experimental profiles. The enhancement of single-strand break (SSB) yields in hypoxic cells by oxygen, para-nitroacetophenone (PNAP), or any of the three nitrofuran derivatives used was well correlated with increased cell killing. Furthermore, reductions in SSB yields for known hydroxyl radical (OH.) scavengers correlates with the reactivities of these compounds toward OH.. This supports the contention that some type of OH.-induced initial lesion, which may ultimately be expressed as an unrepaired or misrepaired double-strand break, constitutes a lethal event. (author)

  2. Formation of double-strand breaks in DNA of γ-irradiated bacteria depending on the function of fast repair processes of DNA single-strand breaks

    International Nuclear Information System (INIS)

    Petrov, S.I.; Gaziev, A.I.

    1980-01-01

    The formation of double-strand breaks in DNA of γ-irradiated ( 60 Co)Ex coli bacteria depending on the function of fast repair processes of DNA single-strand breaks, is investigated. The profiles of sedimentation of DNA Ex coli cells, irradiated at 0-2 deg C in the salt medium and in EDTA-borate buffer, are presented. It is shown that when irradiating cells in EDTA-borate buffer, the output of single- and double strand breaks in DNA is much higher than in the case of their irradiation in the minimum salt medium. The dependence of output of single-strand and double-strand breaks depending on the radiatier doze of E coli cells in the salt medium and EDTA-borate buffer, is studied. The supposition is made on the presence of a regulative interaction between the accumulation of DNA single-breaks and their repair with the formation of double-strand breaks. The functionating of fast and superfast repair processes considerably affects the formation of double-strand breaks in DNA of a bacterium cell. A considerable amount of double-breaks registered immediately after irradiation forms due to a close position of single-strand breaks on the opposite DNA strands

  3. Precision multiloop (PM Design with space closing circles for lingual orthodontics

    Directory of Open Access Journals (Sweden)

    Mugdha P Mankar

    2016-01-01

    Full Text Available The proficiency of ancient orthodontics has been benefitted colossally and is being continually promoted over the present, by use of multiple loop wires designed for correction of dentoalveolar malocclusions. The presented discussion provides an insight into a simple, frictionless biomechanical concept of anterior space closure in lingual orthodontics by means of precision multiloop design with incorporated space closing circles. A multiple loop wire design has been demonstrated where the entire interbracket distance is used as loop area.

  4. Precision multiloop (PM Design) with space closing circles for lingual orthodontics

    OpenAIRE

    Mugdha P Mankar; Achint Chachada; Harish Atram; Avanti Kulkarni

    2016-01-01

    The proficiency of ancient orthodontics has been benefitted colossally and is being continually promoted over the present, by use of multiple loop wires designed for correction of dentoalveolar malocclusions. The presented discussion provides an insight into a simple, frictionless biomechanical concept of anterior space closure in lingual orthodontics by means of precision multiloop design with incorporated space closing circles. A multiple loop wire design has been demonstrated where the ent...

  5. Influência da secção transversa de fios ortodônticos na fricção superficial de braquetes autoligados Influence of the cross-section of orthodontic wires on the surface friction of self-ligating brackets

    Directory of Open Access Journals (Sweden)

    Roberta Buzzoni

    2011-08-01

    Full Text Available OBJETIVO: o objetivo do presente trabalho foi determinar a força de atrito estático entre braquetes de aço inoxidável autoligados com sistema de fechamento resiliente e fios ortodônticos redondos e retangulares do mesmo material. MÉTODOS: empregaram-se 30 braquetes referentes aos caninos superiores divididos em 6 grupos formados por braquetes autoligados Smartclip, In-Ovation R e convencionais Gemini amarrados com ligaduras elásticas. A hipótese testada neste trabalho foi quanto à possibilidade dos braquetes autoligados ativos serem suscetíveis à elevação da força de atrito com o aumento e alteração da secção transversal dos fios ortodônticos. Os ensaios foram realizados com tração de 30s em fios de aço inoxidável 0,020" e 0,019"X0,025" na máquina de ensaios Emic DL 10000, com uma célula de carga de 20 newtons. Cada conjunto braquete/fio foi responsável pela geração de quatro corpos de prova, totalizando 120 leituras. As comparações entre as médias foram realizadas através da Análise de Variância (one way ANOVA com correções pelo coeficiente de Bonferroni. RESULTADOS E CONCLUSÃO: os braquetes autoligados apresentaram maior força de atrito do que os braquetes convencionais amarrados com ligaduras elásticas. O grupo Smartclip foi o mais efetivo no controle do atrito (pOBJECTIVES: The purpose of this study was to assess the surface friction produced by self-ligating stainless steel brackets equipped with a resilient closure system and compare the friction generated during traction of round and rectangular orthodontic wires made from the same material. METHODS: Thirty maxillary canine brackets were divided into six groups comprising SmartClip and In-Ovation R self-ligating brackets, and conventional Gemini brackets tied with elastomeric ligatures. This investigation tested the hypothesis that self-ligating brackets are susceptible to increases in friction that are commensurate with increases and changes in the

  6. Clinicians' Choices in Selecting Orthodontic Archwires

    Directory of Open Access Journals (Sweden)

    Silvia-Izabella Pop

    2013-08-01

    Full Text Available Objective: The aim of this study was to assess the choices made by clinicians in selecting archwires during the initial, intermediate and final stages of orthodontic treatment with fixed appliances. Methods: We carried out a questionnaire-based study at the Orthodontics and Pedodontics Clinic Târgu Mureș, between March 2012 and September 2012. The questionnaires consisted of two parts: the first included questions related to the dimension, alloy used in fabrication, section (round or rectangular and manufacturer of the archwires used by the orthodontists in their orthodontic practice, the second part was concerned with their personal opinion about the physical properties and disadvantages of the archwires. Results: From a total number of 90 distributed questionnaires, 62 were returned. The majority of clinicians are using stainless steel (SS and nickel-titanium alloy (NiTi wires in their fixed orthodontic treatments, very few are using beta-titanium (Beta Ti, copper nickel-titanium (Co- NiTi and esthetic archwires. The preferred dimension seem to be 0.022 inches in the appliance system. Regarding the wire dimensions, 0.014, 0.016 inch wires are mostly used from the round section group and 0.016 × 0.022 inch, 0.017 × 0.025 inch from the rectangular ones. Conclusions: There is a general lack of agreement between the clinicians surveyed regarding the properties of an ideal archwire and the disadvantages of the used wires. The most frequently used alloys seemed to be the SS and NiTi

  7. [The prehistory of orthodontics].

    Science.gov (United States)

    Philippe, Julien

    2015-06-01

    Orthodontics came into being in 1728. Previously, practitioners were at a loss when confronted with crooked teeth. A Latin philosopher had an ingenious flash of orthodontic inspiration. Other authors were content to either extract the malposed teeth or to modify their shape. However, interest in an approach to preventive orthodontics had now begun. © EDP Sciences, SFODF, 2015.

  8. Pelepasan ion nikel dan kromium kawat Australia dan stainless steel dalam saliva buatan (The release of nickel and chromium ions from Australian wire and stainless steel in artificial saliva)

    OpenAIRE

    Nolista Indah Rasyid; Pinandi Sri Pudyani; JCP Heryumani

    2014-01-01

    Background: Fixed orthodontic treatment needs several types of wire to produce biomechanical force to move teeth. The use orthodontic wire within the mouth interacts with saliva, causing the release of nickel and chromium ions. Purpose: The study was aimed to examine the effect of immersion time in artificial saliva between special type of Australian wire and stainless steel on the release of nickel and chromium ions. Methods: Thirty special type Australian wires and 30 stainless steel wires ...

  9. Fixed orthodontic appliances in the management of severe dental trauma in mixed dentition: a case report.

    Science.gov (United States)

    Ebrahim, Fouad-Hassan; Kulkarni, Gajanan

    2013-01-01

    We describe a case of complex trauma to the early mixed dentition in which tooth avulsion, intrusion, extrusion and lateral luxation were managed effectively using a fixed, non-rigid orthodontic splint after treatment with a traditional wire-composite splint had failed. The use of orthodontic brackets and flexible wires provided several advantages, such as the ability to splint severely malpositioned teeth; easy assessment without removing the splint; slow, gentle repositioning of traumatized teeth; and gradual reestablishment of the arch form allowing for ease of future prosthodontic rehabilitation. Therefore, orthodontic appliances should be considered as a viable option for managing complex dental trauma.

  10. Dental extrusion with orthodontic miniscrew anchorage: a case report describing a modified method.

    Science.gov (United States)

    Horliana, Ricardo Fidos; Horliana, Anna Carolina Ratto Tempestini; Wuo, Alexandre do Vale; Perez, Flávio Eduardo Guillin; Abrão, Jorge

    2015-01-01

    In recent years, the skeletal anchorage through miniscrews has expanded the treatment options in orthodontics (Yamaguchi et al., 2012). We hereby present a modified method for tooth extrusion for cases where crown-lengthening surgery is contraindicated for aesthetic reasons. This modified method uses three orthodontic appliances: a mini-implant, an orthodontic wire, and a bracket. The aim of this case report was to increase the length of the clinical crown of a fractured tooth (tooth 23) by means of an orthodontic extrusion with the modified method of Roth and Diedrich.

  11. A novel biomechanical model assessing continuous orthodontic archwire activation

    Science.gov (United States)

    Canales, Christopher; Larson, Matthew; Grauer, Dan; Sheats, Rose; Stevens, Clarke; Ko, Ching-Chang

    2013-01-01

    Objective The biomechanics of a continuous archwire inserted into multiple orthodontic brackets is poorly understood. The purpose of this research was to apply the birth-death technique to simulate insertion of an orthodontic wire and consequent transfer of forces to the dentition in an anatomically accurate model. Methods A digital model containing the maxillary dentition, periodontal ligament (PDL), and surrounding bone was constructed from human computerized tomography data. Virtual brackets were placed on four teeth (central and lateral incisors, canine and first premolar), and a steel archwire (0.019″ × 0.025″) with a 0.5 mm step bend to intrude the lateral incisor was virtually inserted into the bracket slots. Forces applied to the dentition and surrounding structures were simulated utilizing the birth-death technique. Results The goal of simulating a complete bracket-wire system on accurate anatomy including multiple teeth was achieved. Orthodontic force delivered by the wire-bracket interaction was: central incisor 19.1 N, lateral incisor 21.9 N, and canine 19.9 N. Loading the model with equivalent point forces showed a different stress distribution in the PDL. Conclusions The birth-death technique proved to be a useful biomechanical simulation method for placement of a continuous archwire in orthodontic brackets. The ability to view the stress distribution throughout proper anatomy and appliances advances understanding of orthodontic biomechanics. PMID:23374936

  12. Histone H3.3 promotes IgV gene diversification by?enhancing formation of AID?accessible single?stranded DNA

    OpenAIRE

    Romanello, Marina; Schiavone, Davide; Frey, Alexander; Sale, Julian E

    2016-01-01

    Abstract Immunoglobulin diversification is driven by activation?induced deaminase (AID), which converts cytidine to uracil within the Ig variable (IgV) regions. Central to the recruitment of AID to the IgV genes are factors that regulate the generation of single?stranded DNA (ssDNA), the enzymatic substrate of AID. Here, we report that chicken DT40 cells lacking variant histone H3.3 exhibit reduced IgV sequence diversification. We show that this results from impairment of the ability of AID t...

  13. Intramolecular binding mode of the C-terminus of Escherichia coli single-stranded DNA binding protein determined by nuclear magnetic resonance spectroscopy

    OpenAIRE

    Shishmarev, Dmitry; Wang, Yao; Mason, Claire E.; Su, Xun-Cheng; Oakley, Aaron J.; Graham, Bim; Huber, Thomas; Dixon, Nicholas E.; Otting, Gottfried

    2013-01-01

    Single-stranded DNA (ssDNA) binding protein (SSB) is an essential protein to protect ssDNA and recruit specific ssDNA-processing proteins. Escherichia coli SSB forms a tetramer at neutral pH, comprising a structurally well-defined ssDNA binding domain (OB-domain) and a disordered C-terminal domain (C-domain) of ∼64 amino acid residues. The C-terminal eight-residue segment of SSB (C-peptide) has been shown to interact with the OB-domain, but crystal structures failed to reveal any electron den...

  14. Data for increase of Lymantria dispar male survival after topical application of single-stranded RING domain fragment of IAP-3 gene of its nuclear polyhedrosis virus

    Science.gov (United States)

    Oberemok, Volodymyr V.; Laikova, Kateryna V.; Zaitsev, Aleksei S.; Gushchin, Vladimir A.; Skorokhod, Oleksii A.

    2016-01-01

    This data article is related to the research article entitled “The RING for gypsy moth control: topical application of fragment of its nuclear polyhedrosis virus anti-apoptosis gene as insecticide” [1]. This article reports on significantly higher survival of gypsy moth Lymantria dispar male individuals in response to topical application of single-stranded DNA, based on RING (really interesting new gene) domain fragment of LdMNPV (L. dispar multicapsid nuclear polyhedrosis virus) IAP-3 (inhibitor of apoptosis) gene and acted as DNA insecticide. PMID:27054151

  15. Complementarily addressed modification and cleavage of a single-stranded fragment of DNA with the aid of alkylating derivatives of oligonucleotides

    International Nuclear Information System (INIS)

    Brosalina, E.B.; Vlasov, V.V.; Kutyavin, I.V.; Mamaev, S.V.; Pletnev, A.G.; Podyminogin, M.A.

    1986-01-01

    The chemical modification of a 303-nucleotide single-stranded fragment of DNA by alkylating oligonucleotide derivatives bearing 4-[N-methyl-N-(2-chloroethyl)amino]benzyl groups in the 5'-terminal phosphate of the 3'-terminal ribose residue has been investigated. It has been shown that under the conditions of the formation of a complex with the DNA fragment both types of derivatives specifically alkylate nucleotides of the DNA fragments that are located directly adjacent to the sections complementary to the oligonucleotides bearing the reactive groups. Alkylation takes place with a high efficiency, and the DNA fragment can be cleaved specifically at the position of the alkylated nucleotides

  16. Role of DNA repair in repair of cytogenetic damages. Contribution of repair of single-strand DNA breaks to cytogenetic damages repair

    International Nuclear Information System (INIS)

    Rozanova, O.M.; Zaichkina, S.I.; Aptikaev, G.F.; Ganassi, E.Eh.

    1989-01-01

    The comparison was made between the results of the effect of poly(ADP-ribosylation) ingibitors (e.g. nicotinamide and 3-aminobenzamide) and a chromatin proteinase ingibitor, phenylmethylsulfonylfluoride, on the cytogenetic damages repair, by a micronuclear test, and DNA repair in Chinese hamster fibroblasts. The values of the repair half-periods (5-7 min for the cytogenetic damages and 5 min for the rapidly repaired DNA damages) and a similar modyfying effect with regard to radiation cytogenetic damages and kynetics of DNA damages repair were found to be close. This confirms the contribution of repair of DNA single-strand breaks in the initiation of structural damages to chromosomes

  17. [Evaluation of three dimensional orthodontic force produced by magnet of fix appliance].

    Science.gov (United States)

    Dai, Xin; Hou, Zhi-ming; Yao, Ge; Wen, Jing-long

    2008-12-01

    To analyze the feature and magnitude of three dimensional orthodontic force produced by the magnet of fix appliance. Forces detected by universal fatigue test system included the attractive and repulsive,the inclined and rotated orthodontic forces of two magnets in different air gaps, and the integrated inclined and rotated orthodontic forces of two magnets and NiTi wire. The attractive and repulsive forces of two magnets were 4.68 to 0.45 N and 3.00 to 0.40 N respectively in the air gaps of 0 to 5 mm. The inclined orthodontic forces were 1.54 to 1.67 N, 0.63 to 0.69 N, 0.47 to 0.54 N when the magnets were vertically inclined 10 degrees to 40 degrees in the air gaps of 0, 1, 2mm. The rotated orthodontic forces were 0.97 to 1.32 N, 0.53 to 0.59 N, 0.39 to 0.48 N when the magnets were horizontally rotated 10 degrees to 40 degrees in the air gaps of 0, 1, 2mm. The integrated orthodontic force of two magnets and 0.014-inch NiTi wire was 0.32 to 0.5 N when the magnets was vertically inclined 10 degrees to 40 degrees in the air gap of 4 mm. The integrated orthodontic force of two magnets and 0.012-inch NiTi wire was 0.32 to 0.39 N when the magnets were horizontally rotated 10 degrees to 40 degrees in the air gap of 3 mm. Magnets made into orthodontic brackets to some extent could replace the mechanical orthodontic force produced by orthodontic wires and elastics.

  18. Effect of vitamin E on cytotoxicity, DNA single strand breaks, chromosomal aberrations, and mutation in Chinese hamster V-79 cells exposed to ultraviolet-B light

    International Nuclear Information System (INIS)

    Sugiyama, M.; Tsuzuki, K.; Matsumoto, K.; Ogura, R.

    1992-01-01

    The effect of pretreatment with vitamin E on cytotoxicity, DNA single strand breaks, and chromosomal aberrations as well as on mutation induced by ultraviolet-B light (UV-B) was investigated in Chinese hamster V-79 cells. Cellular pretreatment with non-toxic levels of 25 μM α-tocopherol succinate (vitamin E) for 24h prior to exposure resulted in a 10-fold increase in cellular levels of α-tocopherol. Using a colony-forming assay, this pretreatment decreased the cytotoxicity of UV-B light. However, alkaline elution assays demonstrated that pretreatment with vitamin E did not affect the number of DNA single strand breaks caused by UV-B light. UV-B exposure produced a dose-dependent induction of chromosomal aberrations and mutations at the HGPRT locus, and neither of these actions of UV-B was influenced by pretreatment with the vitamin. These results suggest that vitamin E protects cells from UV-B-induced cytotoxicity, possibly through its ability to scavenge free radicals. The genotoxicity induced by UV-B light may not correlate directly with the cytotoxic action of this wavelength region in sunlight. (author)

  19. Formation of DNA single-strand breaks by near-ultraviolet and gamma-rays in normal and Bloom's syndrome skin fibroblasts

    International Nuclear Information System (INIS)

    Hirschi, M.; Netrawali, M.S.; Remsen, J.F.; Cerutti, P.A.

    1981-01-01

    The formation of single-strand breaks by near-ultraviolet light at 313 nm and by aerobic gamma-rays was compared for skin fibroblast monolayer cultures from 4 normal donors (NF) and 8 patients with Bloom's syndrome (BS) by the alkaline elution method. In 6 of 8 BS strains, the number of breaks induced by near-ultraviolet light, 2.25 kJ/sq m, at 0 degrees was comparable to NF, while elevated breakage was observed in BS strains HG 369 and HG 916. Breakage frequencies were increased substantially in 6 of 8 BS strains relative to NF when the near-ultraviolet light exposure was at 37 degrees. BS strain GM 2520 represents an exception since normal breakage frequencies were induced both at 0 degrees and 37 degrees. Aerobic gamma-rays (75 R) induced comparable numbers of single-strand breaks in BS and NF strains at 0 degrees. The breakage frequencies were reduced an average of 17% in NF when the same dose was given at 30 degrees followed by 6 min incubation. Under the same conditions, the breakage frequencies were on the average reduced by 42% relative to 0 degrees in the BS strains, indicating that they possess normal or possibly slightly increased capacities for the rejoining of gamma-ray-induced breaks

  20. Histone H3.3 promotes IgV gene diversification by enhancing formation of AID-accessible single-stranded DNA.

    Science.gov (United States)

    Romanello, Marina; Schiavone, Davide; Frey, Alexander; Sale, Julian E

    2016-07-01

    Immunoglobulin diversification is driven by activation-induced deaminase (AID), which converts cytidine to uracil within the Ig variable (IgV) regions. Central to the recruitment of AID to the IgV genes are factors that regulate the generation of single-stranded DNA (ssDNA), the enzymatic substrate of AID Here, we report that chicken DT40 cells lacking variant histone H3.3 exhibit reduced IgV sequence diversification. We show that this results from impairment of the ability of AID to access the IgV genes due to reduced formation of ssDNA during IgV transcription. Loss of H3.3 also diminishes IgV R-loop formation. However, reducing IgV R-loops by RNase HI overexpression in wild-type cells does not affect IgV diversification, showing that these structures are not necessary intermediates for AID access. Importantly, the reduction in the formation of AID-accessible ssDNA in cells lacking H3.3 is independent of any effect on the level of transcription or the kinetics of RNAPII elongation, suggesting the presence of H3.3 in the nucleosomes of the IgV genes increases the chances of the IgV DNA becoming single-stranded, thereby creating an effective AID substrate. © 2016 MRC Laboratory of Molecular Biology. Published under the terms of the CC BY 4.0 license.

  1. Dynamics of water around the complex structures formed between the KH domains of far upstream element binding protein and single-stranded DNA molecules

    Energy Technology Data Exchange (ETDEWEB)

    Chakraborty, Kaushik; Bandyopadhyay, Sanjoy, E-mail: sanjoy@chem.iitkgp.ernet.in [Molecular Modeling Laboratory, Department of Chemistry, Indian Institute of Technology, Kharagpur 721302 (India)

    2015-07-28

    Single-stranded DNA (ss-DNA) binding proteins specifically bind to the single-stranded regions of the DNA and protect it from premature annealing, thereby stabilizing the DNA structure. We have carried out atomistic molecular dynamics simulations of the aqueous solutions of two DNA binding K homology (KH) domains (KH3 and KH4) of the far upstream element binding protein complexed with two short ss-DNA segments. Attempts have been made to explore the influence of the formation of such complex structures on the microscopic dynamics and hydrogen bond properties of the interfacial water molecules. It is found that the water molecules involved in bridging the ss-DNA segments and the protein domains form a highly constrained thin layer with extremely retarded mobility. These water molecules play important roles in freezing the conformational oscillations of the ss-DNA oligomers and thereby forming rigid complex structures. Further, it is demonstrated that the effect of complexation on the slow long-time relaxations of hydrogen bonds at the interface is correlated with hindered motions of the surrounding water molecules. Importantly, it is observed that the highly restricted motions of the water molecules bridging the protein and the DNA components in the complexed forms originate from more frequent hydrogen bond reformations.

  2. Replication of UV-irradiated single-stranded DNA by DNA polymerase III holoenzyme of Escherichia coli: evidence for bypass of pyrimidine photodimers

    International Nuclear Information System (INIS)

    Livneh, Z.

    1986-01-01

    Replication of UV-irradiated circular single-stranded phage M13 DNA by Escherichia coli RNA polymerase (EC 2.7.7.6) and DNA polymerase III holoenzyme (EC 2.7.7.7) in the presence of single-stranded DNA binding protein yielded full-length as well as partially replicated products. A similar result was obtained with phage G4 DNA primed with E. coli DNA primase, and phage phi X174 DNA primed with a synthetic oligonucleotide. The fraction of full-length DNA was several orders of magnitude higher than predicted if pyrimidine photodimers were to constitute absolute blocks to DNA replication. Recent models have suggested that pyrimidine photodimers are absolute blocks to DNA replication and that SOS-induced proteins are required to allow their bypass. Our results demonstrate that, under in vitro replication conditions, E. coli DNA polymerase III holoenzyme can insert nucleotides opposite pyrimidine dimers to a significant extent, even in the absence of SOS-induced proteins

  3. On-site detection of Phytophthora spp.—single-stranded target DNA as the limiting factor to improve on-chip hybridization

    International Nuclear Information System (INIS)

    Schwenkbier, Lydia; Pollok, Sibyll; Popp, Jürgen; Weber, Karina; König, Stephan; Wagner, Stefan; Werres, Sabine; Weber, Jörg; Hentschel, Martin

    2014-01-01

    We report on a lab-on-a-chip approach for on-site detection of Phytophthora species that allows visual signal readout. The results demonstrate the significance of single-stranded DNA (ssDNA) generation in terms of improving the intensity of the hybridization signal and to improve the reliability of the method. Conventional PCR with subsequent heat denaturation, sodium hydroxide-based denaturation, lambda exonuclease digestion and two asymmetric PCR methods were investigated for the species P. fragariae, P. kernoviae, and P. ramorum. The positioning of the capture probe within the amplified yeast GTP-binding protein (YPT1) target DNA was also of interest because it significantly influences the intensity of the signal. Statistical tests were used to validate the impact of the ssDNA generation methods and the capture-target probe position. The single-stranded target DNA generated by Linear-After-The-Exponential PCR (LATE-PCR) was found to produce signal intensities comparable to post-PCR exonuclease treatment. The LATE-PCR is the best method for the on-site detection of Phytophthora because the enzymatic digestion after PCR is more laborious and time-consuming. (author)

  4. Ca2+ improves organization of single-stranded DNA bases in human Rad51 filament, explaining stimulatory effect on gene recombination.

    KAUST Repository

    Fornander, Louise H

    2012-02-22

    Human RAD51 protein (HsRad51) catalyses the DNA strand exchange reaction for homologous recombination. To clarify the molecular mechanism of the reaction in vitro being more effective in the presence of Ca(2+) than of Mg(2+), we have investigated the effect of these ions on the structure of HsRad51 filament complexes with single- and double-stranded DNA, the reaction intermediates. Flow linear dichroism spectroscopy shows that the two ionic conditions induce significantly different structures in the HsRad51/single-stranded DNA complex, while the HsRad51/double-stranded DNA complex does not demonstrate this ionic dependence. In the HsRad51/single-stranded DNA filament, the primary intermediate of the strand exchange reaction, ATP/Ca(2+) induces an ordered conformation of DNA, with preferentially perpendicular orientation of nucleobases relative to the filament axis, while the presence of ATP/Mg(2+), ADP/Mg(2+) or ADP/Ca(2+) does not. A high strand exchange activity is observed for the filament formed with ATP/Ca(2+), whereas the other filaments exhibit lower activity. Molecular modelling suggests that the structural variation is caused by the divalent cation interfering with the L2 loop close to the DNA-binding site. It is proposed that the larger Ca(2+) stabilizes the loop conformation and thereby the protein-DNA interaction. A tight binding of DNA, with bases perpendicularly oriented, could facilitate strand exchange.

  5. Chromosomal aberrations and deoxyribonucleic acid single-strand breaks in adipose-derived stem cells during long-term expansion in vitro.

    Science.gov (United States)

    Froelich, Katrin; Mickler, Johannes; Steusloff, Gudrun; Technau, Antje; Ramos Tirado, Mario; Scherzed, Agmal; Hackenberg, Stephan; Radeloff, Andreas; Hagen, Rudolf; Kleinsasser, Norbert

    2013-07-01

    Adipose-derived stem cells (ASCs) are a promising mesenchymal cell source for tissue engineering approaches. To obtain an adequate cell amount, in vitro expansion of the cells may be required in some cases. To monitor potential contraindications for therapeutic applications in humans, DNA strand breaks and chromosomal aberrations in ASCs during in vitro expansion were examined. After isolation of ASC from human lipoaspirates of seven patients, in vitro expansion over 10 passages was performed. Cells from passages 1, 2, 3, 5 and 10 were used for the alkaline single-cell microgel electrophoresis (comet) assay to detect DNA single-strand breaks and alkali labile as well as incomplete excision repair sites. Chromosomal changes were examined by means of the chromosomal aberration test. During in vitro expansion, ASC showed no DNA single-strand breaks in the comet assay. With the chromosomal aberration test, however, a significant increase in chromosomal aberrations were detected. The study showed that although no DNA fragmentation could be determined, the safety of ASC cannot be ensured with respect to chromosome stability during in vitro expansion. Thus, reliable analyses for detecting ASC populations, which accumulate chromosomal aberrations or even undergo malignant transformation during extensive in vitro expansion, must be implemented as part of the safety evaluation of these cells for stem cell-based therapy. Copyright © 2013 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  6. A single-strand specific lesion drives MMS-induced hyper-mutability at a double-strand break in yeast.

    Science.gov (United States)

    Yang, Yong; Gordenin, Dmitry A; Resnick, Michael A

    2010-08-05

    Localized hyper-mutability (LHM) can be important in evolution, immunity, and genetic diseases. We previously reported that single-strand DNA (ssDNA) can be an important source of damage-induced LHM in yeast. Here, we establish that the generation of LHM by methyl methanesulfonate (MMS) during repair of a chromosomal double-strand break (DSB) can result in over 0.2 mutations/kb, which is approximately 20,000-fold higher than the MMS-induced mutation density without a DSB. The MMS-induced mutations associated with DSB repair were primarily due to substitutions via translesion DNA synthesis at damaged cytosines, even though there are nearly 10 times more MMS-induced lesions at other bases. Based on this mutation bias, the promutagenic lesion dominating LHM is likely 3-methylcytosine, which is single-strand specific. Thus, the dramatic increase in mutagenesis at a DSB is concluded to result primarily from the generation of non-repairable lesions in ssDNA associated with DSB repair along with efficient induction of highly mutagenic ssDNA-specific lesions. These findings with MMS-induced LHM have broad biological implications for unrepaired damage generated in ssDNA and possibly ssRNA. Published by Elsevier B.V.

  7. Intracellular generation of single-strand template increases the knock-in efficiency by combining CRISPR/Cas9 with AAV.

    Science.gov (United States)

    Xiao, Qing; Min, Taishan; Ma, Shuangping; Hu, Lingna; Chen, Hongyan; Lu, Daru

    2018-04-18

    Targeted integration of transgenes facilitates functional genomic research and holds prospect for gene therapy. The established microhomology-mediated end-joining (MMEJ)-based strategy leads to the precise gene knock-in with easily constructed donor, yet the limited efficiency remains to be further improved. Here, we show that single-strand DNA (ssDNA) donor contributes to efficient increase of knock-in efficiency and establishes a method to achieve the intracellular linearization of long ssDNA donor. We identified that the CRISPR/Cas9 system is responsible for breaking double-strand DNA (dsDNA) of palindromic structure in inverted terminal repeats (ITRs) region of recombinant adeno-associated virus (AAV), leading to the inhibition of viral second-strand DNA synthesis. Combing Cas9 plasmids targeting genome and ITR with AAV donor delivery, the precise knock-in of gene cassette was achieved, with 13-14% of the donor insertion events being mediated by MMEJ in HEK 293T cells. This study describes a novel method to integrate large single-strand transgene cassettes into the genomes, increasing knock-in efficiency by 13.6-19.5-fold relative to conventional AAV-mediated method. It also provides a comprehensive solution to the challenges of complicated production and difficult delivery with large exogenous fragments.

  8. Wire Chamber

    CERN Multimedia

    Magnetoscriptive readout wire chamber. Multi-wire detectors contain layers of positively and negatively charged wires enclosed in a chamber full of gas. A charged particle passing through the chamber knocks negatively charged electrons out of atoms in the gas, leaving behind positive ions. The electrons are pulled towards the positively charged wires. They collide with other atoms on the way, producing an avalanche of electrons and ions. The movement of these electrons and ions induces an electric pulse in the wires which is collected by fast electronics. The size of the pulse is proportional to the energy loss of the original particle.

  9. Wire chamber

    CERN Multimedia

    1967-01-01

    Magnetoscriptive readout wire chamber.Multi-wire detectors contain layers of positively and negatively charged wires enclosed in a chamber full of gas. A charged particle passing through the chamber knocks negatively charged electrons out of atoms in the gas, leaving behind positive ions. The electrons are pulled towards the positively charged wires. They collide with other atoms on the way, producing an avalanche of electrons and ions. The movement of these electrons and ions induces an electric pulse in the wires which is collected by fast electronics. The size of the pulse is proportional to the energy loss of the original particle.

  10. wire chamber

    CERN Multimedia

    Proportional multi-wire chamber. Multi-wire detectors contain layers of positively and negatively charged wires enclosed in a chamber full of gas. A charged particle passing through the chamber knocks negatively charged electrons out of atoms in the gas, leaving behind positive ions. The electrons are pulled towards the positively charged wires. They collide with other atoms on the way, producing an avalanche of electrons and ions. The movement of these electrons and ions induces an electric pulse in the wires which is collected by fast electronics. The size of the pulse is proportional to the energy loss of the original particle. Proportional wire chambers allow a much quicker reading than the optical or magnetoscriptive readout wire chambers.

  11. Scaleup of powder metallurgy processed Nb-Al multifilamentary wire

    International Nuclear Information System (INIS)

    Thieme, C.; Foner, S.; Otubo, J.; Pourrahimi, S.; Schwartz, B.; Zhang, H.

    1983-01-01

    Power metallurgy processed Nb-Al superconducting wires were fabricated from billets up to 45 mm o.d. with nominal areal reduction ratios, R, up to 2 X 10 5 , Nb powder sizes from 40 to 300 μm from various sources, Al powder sizes from 9 to 75 μm, Al concentrations from 3 to 25 wt % Al and with a wide range of heat treatments. All the compacts used tap density powder in a Cu tube and swaging and/or rod rolling and subsequent wire drawing. Both single strand and bundled wires were made. Overall critical current densities, J /SUB c/, of 2 X 10 4 A/cm 2 at 14 T and 10 4 A/cm 2 at 16 T were achieved for 6 to 8 wt % Al in Nb

  12. Allergy and orthodontics

    Science.gov (United States)

    Chakravarthi, Sunitha; Padmanabhan, Sridevi; Chitharanjan, Arun B.

    2012-01-01

    The aim of this paper is to review the current literature on allergy in orthodontics and to identify the predisposing factors and the implications of the allergic reaction in the management of patients during orthodontic treatment. A computerized literature search was conducted in PubMed for articles published on allergy in relation to orthodontics. The MeSH term used was allergy and orthodontics. Allergic response to alloys in orthodontics, particularly nickel, has been extensively studied and several case reports of nickel-induced contact dermatitis have been documented. Current evidence suggests that the most common allergic reaction reported in orthodontics is related to nickel in orthodontic appliances and allergic response is more common in women due to a previous sensitizing exposure from nickel in jewellery. Studies have implicated allergy in the etiology of hypo-dontia. It has also been considered as a high-risk factor for development of extensive root resorption during the course of orthodontic treatment. This review discusses the relationship and implications of allergy in orthodontics. PMID:24987632

  13. Absolute determination of single-stranded and self-complementary adeno-associated viral vector genome titers by droplet digital PCR.

    Science.gov (United States)

    Lock, Martin; Alvira, Mauricio R; Chen, Shu-Jen; Wilson, James M

    2014-04-01

    Accurate titration of adeno-associated viral (AAV) vector genome copies is critical for ensuring correct and reproducible dosing in both preclinical and clinical settings. Quantitative PCR (qPCR) is the current method of choice for titrating AAV genomes because of the simplicity, accuracy, and robustness of the assay. However, issues with qPCR-based determination of self-complementary AAV vector genome titers, due to primer-probe exclusion through genome self-annealing or through packaging of prematurely terminated defective interfering (DI) genomes, have been reported. Alternative qPCR, gel-based, or Southern blotting titering methods have been designed to overcome these issues but may represent a backward step from standard qPCR methods in terms of simplicity, robustness, and precision. Droplet digital PCR (ddPCR) is a new PCR technique that directly quantifies DNA copies with an unparalleled degree of precision and without the need for a standard curve or for a high degree of amplification efficiency; all properties that lend themselves to the accurate quantification of both single-stranded and self-complementary AAV genomes. Here we compare a ddPCR-based AAV genome titer assay with a standard and an optimized qPCR assay for the titration of both single-stranded and self-complementary AAV genomes. We demonstrate absolute quantification of single-stranded AAV vector genomes by ddPCR with up to 4-fold increases in titer over a standard qPCR titration but with equivalent readout to an optimized qPCR assay. In the case of self-complementary vectors, ddPCR titers were on average 5-, 1.9-, and 2.3-fold higher than those determined by standard qPCR, optimized qPCR, and agarose gel assays, respectively. Droplet digital PCR-based genome titering was superior to qPCR in terms of both intra- and interassay precision and is more resistant to PCR inhibitors, a desirable feature for in-process monitoring of early-stage vector production and for vector genome biodistribution

  14. Comparação de parâmetros periodontais após utilização de contenção ortodôntica com fio trançado e contenção modificada Comparison of periodontal parameters after the use of orthodontic multi-stranded wire retainers and modified retainers

    Directory of Open Access Journals (Sweden)

    Marlice Azoia Lukiantchuki

    2011-08-01

    Full Text Available OBJETIVO: o presente estudo teve como objetivo comparar dois tipos de contenções ortodônticas fixas, em relação a parâmetros periodontais estabelecidos. A contenção com fio trançado é a mais comumente utilizada e a contenção modificada apresenta dobras que têm a finalidade de permitir o livre acesso do fio dental às áreas interproximais. MÉTODOS: para esse estudo cruzado, foram selecionados 12 voluntários que utilizaram, por 6 meses, A Contenção com fio trançado e B Contenção modificada - sendo essas fixadas em todos os dentes do segmento anterior. Após esse período experimental, foram feitas as seguintes avaliações: Índice de Placa Dentária, Índice Gengival, Índice de Cálculo Dentário e Índice de Cálculo ao longo do fio de contenção. Os voluntários também responderam a um questionário com relação à utilização, conforto e higienização das contenções. RESULTADOS: foi observado que o índice de placa e o índice gengival foram maiores nas faces linguais (pOBJECTIVE: The objective of the present study was to compare two types of fixed orthodontic retainers (a multi-stranded wire retainer and a modified retainer in relation to established periodontal parameters. The multi-stranded wire retainer is commonly used, and the modified retainer has bends to enable free access of dental floss to interproximal areas. METHODS: For this crossover study, 12 volunteers were selected and used the following retainers for six months: (A a multi-stranded wire retainer and (B a modified retainer. Both retainers were fixed to all anterior lower teeth.After this experimental period, the following evaluations were made: Dental Plaque Index, Gingival Index, Dental Calculus Index and Retainer Wire Calculus Index. The volunteers also responded to a questionnaire about the use, comfort and hygiene of the retainers. RESULTS: It was observed that the plaque index and the gingival index were higher on the lingual surface (p<0

  15. wire chamber

    CERN Multimedia

    1985-01-01

    Multi-wire detectors contain layers of positively and negatively charged wires enclosed in a chamber full of gas. A charged particle passing through the chamber knocks negatively charged electrons out of atoms in the gas, leaving behind positive ions. The electrons are pulled towards the positively charged wires. They collide with other atoms on the way, producing an avalanche of electrons and ions. The movement of these electrons and ions induces an electric pulse in the wires which is collected by fast electronics. The size of the pulse is proportional to the energy loss of the original particle.

  16. Wire chamber

    CERN Multimedia

    Multi-wire detectors contain layers of positively and negatively charged wires enclosed in a chamber full of gas. A charged particle passing through the chamber knocks negatively charged electrons out of atoms in the gas, leaving behind positive ions. The electrons are pulled towards the positively charged wires. They collide with other atoms on the way, producing an avalanche of electrons and ions. The movement of these electrons and ions induces an electric pulse in the wires which is collected by fast electronics. The size of the pulse is proportional to the energy loss of the original particle.

  17. wire chamber

    CERN Multimedia

    Multi-wire detectors contain layers of positively and negatively charged wires enclosed in a chamber full of gas. A charged particle passing through the chamber knocks negatively charged electrons out of atoms in the gas, leaving behind positive ions. The electrons are pulled towards the positively charged wires. They collide with other atoms on the way, producing an avalanche of electrons and ions. The movement of these electrons and ions induces an electric pulse in the wires which is collected by fast electronics. The size of the pulse is proportional to the energy loss of the original particle.

  18. wire chamber

    CERN Multimedia

    Was used in ISR (Intersecting Storage Ring) split field magnet experiment. Multi-wire detectors contain layers of positively and negatively charged wires enclosed in a chamber full of gas. A charged particle passing through the chamber knocks negatively charged electrons out of atoms in the gas, leaving behind positive ions. The electrons are pulled towards the positively charged wires. They collide with other atoms on the way, producing an avalanche of electrons and ions. The movement of these electrons and ions induces an electric pulse in the wires which is collected by fast electronics. The size of the pulse is proportional to the energy loss of the original particle.

  19. [Preprosthetic orthodontic treatments

    NARCIS (Netherlands)

    Prahl-Andersen, B.; Prahl, C.; Baat, C. de; Creugers, N.H.J.

    2014-01-01

    The objective of a preprosthetic orthodontic treatment is to position the teeth in such a way that a treatment with (fixed) dental prostheses is made possible or simplified or to affect the result of this treatment positively. Conceivable preprosthetic orthodontic treatments are: correcting primary

  20. Synthesis of a wild-type and three mutant Cucurbita maxima trypsin inhibitor-encoding genes by a single-strand approach.

    Science.gov (United States)

    Botes, D P; Qobose, M D; Corfield, V A

    1991-09-15

    A single-strand approach to gene assembly, based on a modification of an in vitro complementary oligodeoxyribonucleotide template-directed ligation of the desired sequence to a linearized vector [Chen et al., Nucleic Acids Res. 18 (1990) 871-878], is described. The gene coding for the wild-type Cucurbita maxima trypsin inhibitor of 29 amino acid residues [Bode et al., FEBS Lett. 242 (1989) 285-292], as well as three mutant forms of the gene, in which two of the three disulfide bonds have been replaced singly or as a pair, have been synthesized in a single synthesis run with minimal manual intervention. Subsequent to ligation to pUC9 and in vivo gapped duplex repair by Escherichia coli, their sequences have been verified.

  1. Calibration of denaturing agarose gels for molecular weight estimation of DNA: size determination of the single-stranded genomes of parvoviruses

    Energy Technology Data Exchange (ETDEWEB)

    Snyder, C.E. (Oak Ridge National Lab., TN); Schmoyer, R.L.; Bates, R.C.; Mitra, S.

    1982-01-01

    Vertical slab gel electrophoresis of DNA with CH/sub 3/HgOH-containing agarose produces sharp bands whose mobilities are suitable for size estimation of single-stranded DNA containing 600 to 20,000 bases. The relationship of electrophoretic mobility to size of DNA over this range is a smooth, S-shaped function, and an empirical model was developed to express the relationship. The model involves terms in squared and reciprocal mobilities, and produced excellent fit of known standard markers to measured mobilities. It was used to estimate the sizes of six parvovirus DNAs: Kilham rat virus (KRV), H-1, LuIII, and minute virus of mice (MVM) DNAs had molecular weights of 1.66 to 1.70 x 10/sup 6/, while the molecular weight of bovine parvovirus (BPV) DNA was 1.84 x 10/sup 6/ and that of adenoassociated virus (AAV) DNA was 1.52 x 10/sup 6/.

  2. UV light-induced DNA synthesis arrest in HeLa cells is associated with changes in phosphorylation of human single-stranded DNA-binding protein

    International Nuclear Information System (INIS)

    Carty, M.P.; Zernik-Kobak, M.; McGrath, S.; Dixon, K.

    1994-01-01

    We show that DNA replication activity in extracts of human HeLa cells decreases following UV irradiation. Alterations in replication activity in vitro parallel the UV-induced block in cell cycle progression of these cells in culture. UV irradiation also induces specific changes in the pattern of phosphorylation of the 34 kDa subunit of a DNA replication protein, human single-stranded DNA-binding protein (hSSB). The appearance of a hyperphosphorylated form of hSSB correlates with reduced in vitro DNA replication activity in extracts of UV-irradiated cells. Replication activity can be restored to these extracts in vitro by addition of purified hSSB. These results suggest that UV-induced DNA synthesis arrest may be mediated in part through phosphorylation-related alterations in the activity of hSSB, an essential component of the DNA replication apparatus. (Author)

  3. Effects of DNA double-strand and single-strand breaks on intrachromosomal recombination events in cell-cycle-arrested yeast cells

    International Nuclear Information System (INIS)

    Galli, A.; Schiestl, R.H.

    1998-01-01

    Intrachromosomal recombination between repeated elements can result in deletion (DEL recombination) events. We investigated the inducibility of such intrachromosomal recombination events at different stages of the cell cycle and the nature of the primary DNA lesions capable of initiating these events. Two genetic systems were constructed in Saccharomyces cerevisiae that select for DEL recombination events between duplicated alleles of CDC28 and TUB2. We determined effects of double-strand breaks (DSBs) and single-strand breaks (SSBs) between the duplicated alleles on DEL recombination when induced in dividing cells or cells arrested in G1 or G2. Site-specific DSBs and SSBs were produced by overexpression of the I-Sce I endonuclease and the gene II protein (gIIp), respectively. I-Sce I-induced DSBs caused an increase in DEL recombination frequencies in both dividing and cell-cycle-arrested cells, indicating that G1- and G2-arrested cells are capable of completing DSB repair. In contrast, gIIp-induced SSBs caused an increase in DEL recombination frequency only in dividing cells. To further examine these phenomena we used both γ-irradiation, inducing DSBs as its most relevant lesion, and UV, inducing other forms of DNA damage. UV irradiation did not increase DEL recombination frequencies in G1 or G2, whereas γ-rays increased DEL recombination frequencies in both phases. Both forms of radiation, however, induced DEL recombination in dividing cells. The results suggest that DSBsbut not SSBs induce DEL recombination, probably via the single-strand annealing pathway. Further, DSBs in dividing cells may result from the replication of a UV or SSB-damaged template. Alternatively, UV induced events may occur by replication slippage after DNA polymerase pausing in front of the damage. (author)

  4. Localization of specific sequences and DNA single-strand breaks in individual UV-A-irradiated human lymphocytes by COMET FISH

    Science.gov (United States)

    Bock, Claudia; Rapp, Alexander; Dittmar, Heike; Monajembashi, Shamci; Greulich, Karl-Otto

    1999-01-01

    The COMET assay, a single cell electrophoresis technique which allows to separate electrophoretically fractionated DNA according to size has been combined with fluorescence in situ hybridization (FISH) which allows to localize specific genes or gene regions. This combination (COMET FISH) allows the detection of DNA single strand breaks in specific regions of the genome of human lymphocytes at the single cell level. Various types of DNA probes, e.g. centromere-, (alpha) - satellite-, telomere-, whole chromosome-, single copy- and region specific DNA probes have been used to investigate whether the UV-A induced DNA single strand breaks are distributed randomly all over the human genome or induced at specific sites ('hot spots'). In the investigated human peripheral blood lymphocytes all but one centromere reveal low sensitivity for UV-A irradiation (500 kJ/m2), while telomeres are randomly distributed over COMET heads and tails. The human chromosome 1 is fractionated by irradiation, but remains in the COMET head, indicating an only moderate degree of fractionation. Among three tested single copy probes, c- myc, p53 and p58, the p53 gene located on chromosome 17p13.1 and the p58 gene (1p36) appear to be located in UV-A stable regions of the human genome in 95% of 65 investigated lymphocytes. In contrast, the c-myc proto-oncogene (8q24) is found in the COMET tail in 90% of the 27 investigated lymphocytes and thus appears to be more sensitive to UV-A irradiation.

  5. Single-strand conformation polymorphism (SSCP)-based mutation scanning approaches to fingerprint sequence variation in ribosomal DNA of ascaridoid nematodes.

    Science.gov (United States)

    Zhu, X Q; Gasser, R B

    1998-06-01

    In this study, we assessed single-strand conformation polymorphism (SSCP)-based approaches for their capacity to fingerprint sequence variation in ribosomal DNA (rDNA) of ascaridoid nematodes of veterinary and/or human health significance. The second internal transcribed spacer region (ITS-2) of rDNA was utilised as the target region because it is known to provide species-specific markers for this group of parasites. ITS-2 was amplified by PCR from genomic DNA derived from individual parasites and subjected to analysis. Direct SSCP analysis of amplicons from seven taxa (Toxocara vitulorum, Toxocara cati, Toxocara canis, Toxascaris leonina, Baylisascaris procyonis, Ascaris suum and Parascaris equorum) showed that the single-strand (ss) ITS-2 patterns produced allowed their unequivocal identification to species. While no variation in SSCP patterns was detected in the ITS-2 within four species for which multiple samples were available, the method allowed the direct display of four distinct sequence types of ITS-2 among individual worms of T. cati. Comparison of SSCP/sequencing with the methods of dideoxy fingerprinting (ddF) and restriction endonuclease fingerprinting (REF) revealed that also ddF allowed the definition of the four sequence types, whereas REF displayed three of four. The findings indicate the usefulness of the SSCP-based approaches for the identification of ascaridoid nematodes to species, the direct display of sequence variation in rDNA and the detection of population variation. The ability to fingerprint microheterogeneity in ITS-2 rDNA using such approaches also has implications for studying fundamental aspects relating to mutational change in rDNA.

  6. Effect of material variation on the biomechanical behaviour of orthodontic fixed appliances: a finite element analysis.

    Science.gov (United States)

    Papageorgiou, Spyridon N; Keilig, Ludger; Hasan, Istabrak; Jäger, Andreas; Bourauel, Christoph

    2016-06-01

    Biomechanical analysis of orthodontic tooth movement is complex, as many different tissues and appliance components are involved. The aim of this finite element study was to assess the relative effect of material alteration of the various components of the orthodontic appliance on the biomechanical behaviour of tooth movement. A three-dimensional finite element solid model was constructed. The model consisted of a canine, a first, and a second premolar, including the surrounding tooth-supporting structures and fixed appliances. The materials of the orthodontic appliances were alternated between: (1) composite resin or resin-modified glass ionomer cement for the adhesive, (2) steel, titanium, ceramic, or plastic for the bracket, and (3) β-titanium or steel for the wire. After vertical activation of the first premolar by 0.5mm in occlusal direction, stress and strain calculations were performed at the periodontal ligament and the orthodontic appliance. The finite element analysis indicated that strains developed at the periodontal ligament were mainly influenced by the orthodontic wire (up to +63 per cent), followed by the bracket (up to +44 per cent) and the adhesive (up to +4 per cent). As far as developed stresses at the orthodontic appliance are concerned, wire material had the greatest influence (up to +155 per cent), followed by bracket material (up to +148 per cent) and adhesive material (up to +8 per cent). The results of this in silico study need to be validated by in vivo studies before they can be extrapolated to clinical practice. According to the results of this finite element study, all components of the orthodontic fixed appliance, including wire, bracket, and adhesive, seem to influence, to some extent, the biomechanics of tooth movement. © The Author 2015. Published by Oxford University Press on behalf of the European Orthodontic Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  7. Implants for orthodontic anchorage

    Science.gov (United States)

    Zheng, Xiaowen; Sun, Yannan; Zhang, Yimei; Cai, Ting; Sun, Feng; Lin, Jiuxiang

    2018-01-01

    Abstract Implantanchorage continues to receive much attention as an important orthodontic anchorage. Since the development of orthodontic implants, the scope of applications has continued to increase. Although multiple reviews detailing implants have been published, no comprehensive evaluations have been performed. Thus, the purpose of this study was to comprehensively evaluate the effects of implants based on data published in review articles. An electronic search of the Cochrane Library, Medline, Embase, Ebsco and Sicencedirect for reviews with “orthodontic” and “systematic review or meta analysis” in the title, abstract, keywords, or full text was performed. A subsequent manual search was then performed to identify reviews concerning orthodontic implants. A manual search of the orthodontic journals American Journal of Orthodontics and Dentofacial Orthopedics (AJODO), European Journal of Orthodontics (EJO), and Angle Othodontist was also performed. Such systematic reviews that evaluated the efficacy and safety of orthodontic implants were used to indicate success rates and molar movements. A total of 23 reviews were included in the analysis. The quality of each review was assessed using a measurement tool for Assessment of Multiple Systematic Reviews (AMSTAR), and the review chosen to summarize outcomes had a quality score of >6. Most reviews were less than moderate quality. Success rates of implants ranged in a broad scope, and movement of the maxillary first molar was superior with implants compared with traditional anchorage. PMID:29595673

  8. FACTORS ASSOCIATED WITH ODONTOGENIC BACTERAEMIA IN ORTHODONTIC PATIENTS.

    Science.gov (United States)

    Umeh, O D; Sanu, O O; Utomi, I L; Nwaokorie, F O

    2016-01-01

    Various researches have investigated factors associated with the prevalence and intensity of bacteraemia following oral procedures including orthodontic procedures. The aim of this study was to determine the effect of age, gender, plaque and gingival indices on the occurrence of odontogenic bacteraemia following orthodontic treatment procedures. Orthodontic Clinic, Lagos University Teaching Hospital (LUTH), Lagos , Nigeria. Using the consecutive, convenience sampling method, a total of 100 subjects who met the inclusion criteria were recruited for the study and peripheral blood was collected before and again within 2 minutes of completion of orthodontic procedures for microbiologic analysis using the BACTEC automated blood culture system and the lysis filtration methods of blood culturing. The subjects were randomly placed in one of four orthodontic procedures investigated: alginate impression making (Group I), separator placement (Group II), band cementation (Group III) and arch wire change (Group IV). Plaque and gingival indices were assessed using the plaque component of the Simplified Oral Hygiene Index (OHI-S) (Greene & Vermillion) and Modified gingival index (Lobene) respectively before blood collection. Spearman Point bi-serial correlations and logistic regression statistics were used for statistical evaluations at p orthodontic treatment procedures of 16% and 28% were observed respectively using the BACTEC and lysis filtration methods. A statistically significant increase in the prevalence of bateraemia was observed following separator placement (p=0.016). An increase in age, plaque index scores and modified gingival index scores of the subjects were found to be associated with an increase in the prevalence of bacteraemia following orthodontic treatment procedures, with plaque index score showing the strongest correlation. Separator placement was found to induce significantly highest level of bacteraemia. Meticulous oral hygiene practice and the use of 0

  9. Numerical Investigation of Corrugated Wire Mesh Laminate

    Directory of Open Access Journals (Sweden)

    Jeongho Choi

    2013-01-01

    Full Text Available The aim of this work is to develop a numerical model of Corrugated Wire Mesh Laminate (CWML capturing all its complexities such as nonlinear material properties, nonlinear geometry and large deformation behaviour, and frictional behaviour. Development of such a model will facilitate numerical simulation of the mechanical behaviour of the wire mesh structure under various types of loading as well as the variation of the CWML configuration parameters to tailor its mechanical properties to suit the intended application. Starting with a single strand truss model consisting of four waves with a bilinear stress-strain model to represent the plastic behaviour of stainless steel, the finite element model is gradually built up to study single-layer structures with 18 strands of corrugated wire meshes consistency and double- and quadruple-layered laminates with alternating crossply orientations. The compressive behaviour of the CWML model is simulated using contact elements to model friction and is compared to the load-deflection behaviour determined experimentally in uniaxial compression tests. The numerical model of the CWML is then employed to conduct the aim of establishing the upper and lower bounds of stiffness and load capacity achievable by such structures.

  10. Preparation, mechanical, and in vitro properties of glass fiber-reinforced polycarbonate composites for orthodontic application.

    Science.gov (United States)

    Tanimoto, Yasuhiro; Inami, Toshihiro; Yamaguchi, Masaru; Nishiyama, Norihiro; Kasai, Kazutaka

    2015-05-01

    Generally, orthodontic treatment uses metallic wires made from stainless steel, cobalt-chromium-nickel alloy, β-titanium alloy, and nickel-titanium (Ni-Ti) alloy. However, these wires are not esthetically pleasing and may induce allergic or toxic reactions. To correct these issues, in the present study we developed glass-fiber-reinforced plastic (GFRP) orthodontic wires made from polycarbonate and E-glass fiber by using pultrusion. After fabricating these GFRP round wires with a diameter of 0.45 mm (0.018 inch), we examined their mechanical and in vitro properties. To investigate how the glass-fiber diameter affected their physical properties, we prepared GFRP wires of varying diameters (7 and 13 µm). Both the GFRP with 13-µm fibers (GFRP-13) and GFRP with 7 µm fibers (GFRP-7) were more transparent than the metallic orthodontic wires. Flexural strengths of GFRP-13 and GFRP-7 were 690.3 ± 99.2 and 938.1 ± 95.0 MPa, respectively; flexural moduli of GFRP-13 and GFRP-7 were 25.4 ± 4.9 and 34.7 ± 7.7 GPa, respectively. These flexural properties of the GFRP wires were nearly equivalent to those of available Ni-Ti wires. GFRP-7 had better flexural properties than GFRP-13, indicating that the flexural properties of GFRP increase with decreasing fiber diameter. Using thermocycling, we found no significant change in the flexural properties of the GFRPs after 600 or 1,200 cycles. Using a cytotoxicity detection kit, we found that the glass fiber and polycarbonate components comprising the GFRP were not cytotoxic within the limitations of this study. We expect this metal-free GFRP wire composed of polycarbonate and glass fiber to be useful as an esthetically pleasing alternative to current metallic orthodontic wire. © 2014 Wiley Periodicals, Inc.

  11. 21 CFR 872.5410 - Orthodontic appliance and accessories.

    Science.gov (United States)

    2010-04-01

    ... (CONTINUED) MEDICAL DEVICES DENTAL DEVICES Therapeutic Devices § 872.5410 Orthodontic appliance and... device includes the preformed orthodontic band, orthodontic band material, orthodontic elastic band...

  12. Orthodontic management of impacted central incisor: A clinical challenge

    Directory of Open Access Journals (Sweden)

    Amit Kumar Khera

    2017-01-01

    Full Text Available Multiple treatment options are available for patients who have impacted incisor. This paper shows a case in which orthodontic as well as surgical considerations in 10-year-old female child were presented in the management of impacted central incisor. The orthodontic treatment plan included three steps – creation of space, exposure of crown, and forced eruption. A unique and innovative technique for orthodontic traction (0.017 × 0.025 TMA wire with palatal extension was employed to move the maxillary incisor into arch, with minimum injury to neighboring soft tissue. After the successful management of impacted teeth, it is very important to periodically review the periodontal condition and stability.

  13. An orthodontic oral appliance.

    Science.gov (United States)

    Marklund, Marie; Legrell, Per Erik

    2010-11-01

    This pilot study was performed to test the hypothesis that an orthodontic oral appliance (OA) that is designed to work against the backwardly directed forces on the upper incisors may counteract the reduction in overjet from these devices. Thirty patients with normal bites, good oral health, and milder sleep apnea were randomized to treatment with either OAs or orthodontic OAs. Bite changes were evaluated on plaster casts and radiographs and by questionnaires after a mean of 2.4 years in 19 frequent users. Four of nine patients in the orthodontic OA group increased their overjet by > or =0.4 mm, while none of the 10 patients in the OA group experienced that effect. Only the orthodontic OA increases the overjet; this design may therefore be beneficial to patients at risk of negative effects on their bite during OA treatment.

  14. Lingual Orthodontics simplified : Incognito -customization perfected

    Directory of Open Access Journals (Sweden)

    Haroutioun Dedeyan

    2013-01-01

    Full Text Available Esthetic orthodontics is the need of the hour, fuelled by the increasing number of adult patients seeking orthodontic treatment. Alternatives to labial appliance include clear aligners and lingual appliance. Conventional lingual treatment is laborious in terms of laboratory setup and manual dexterity of the operator, coupled with less than optimum treatment results. Customization of the appliance to meet varied requirements of each patient due to highly variable lingual morphology within and amongst patients is the key to successful treatment in lingual orthodontics. Efforts at bracket base customization using intra-oral jigs and laboratory setups are fraught with unavoidable errors affecting treatment outcome. With the advent of computer-aided design and computer-aided manufacturing (CAD/CAM technology the Eldorado of true customization is now a reality. This paper introduces the Incognito Appliance System based on custom-made brackets and custom-made series of pre-bent wires using state of art CAD/CAM manufacturing procedures.

  15. Single Strand Annealing Plays a Major Role in RecA-Independent Recombination between Repeated Sequences in the Radioresistant Deinococcus radiodurans Bacterium.

    Directory of Open Access Journals (Sweden)

    Solenne Ithurbide

    2015-10-01

    Full Text Available The bacterium Deinococcus radiodurans is one of the most radioresistant organisms known. It is able to reconstruct a functional genome from hundreds of radiation-induced chromosomal fragments. Our work aims to highlight the genes involved in recombination between 438 bp direct repeats separated by intervening sequences of various lengths ranging from 1,479 bp to 10,500 bp to restore a functional tetA gene in the presence or absence of radiation-induced DNA double strand breaks. The frequency of spontaneous deletion events between the chromosomal direct repeats were the same in recA+ and in ΔrecA, ΔrecF, and ΔrecO bacteria, whereas recombination between chromosomal and plasmid DNA was shown to be strictly dependent on the RecA and RecF proteins. The presence of mutations in one of the repeated sequence reduced, in a MutS-dependent manner, the frequency of the deletion events. The distance between the repeats did not influence the frequencies of deletion events in recA+ as well in ΔrecA bacteria. The absence of the UvrD protein stimulated the recombination between the direct repeats whereas the absence of the DdrB protein, previously shown to be involved in DNA double strand break repair through a single strand annealing (SSA pathway, strongly reduces the frequency of RecA- (and RecO- independent deletions events. The absence of the DdrB protein also increased the lethal sectoring of cells devoid of RecA or RecO protein. γ-irradiation of recA+ cells increased about 10-fold the frequencies of the deletion events, but at a lesser extend in cells devoid of the DdrB protein. Altogether, our results suggest a major role of single strand annealing in DNA repeat deletion events in bacteria devoid of the RecA protein, and also in recA+ bacteria exposed to ionizing radiation.

  16. Laser diagnostics in orthodontics

    Science.gov (United States)

    Ryzhkova, Anastasia V.; Lebedeva, Nina G.; Sedykh, Alexey V.; Ulyanov, Sergey S.; Lepilin, Alexander V.; Kharish, Natalia A.

    2003-10-01

    The results of statistical analysis of Doppler spectra of intensity fluctuations of light, scattered from mucose membrane of oral cavity of healthy volunteers and patients, abused by the orthodontic diseases, are presented. Analysis of Doppler spectra, obtained from tooth pulp of patients, is carried out. New approach to monitoring of blood microcirculation in orthodontics is suggested. Influence of own noise of measuring system on formation of the speckle-interferometric signal is studied.

  17. Analysis of optimum wire rope configuration for equal unidirectional torsional stiffness for flexible steering shaft

    Directory of Open Access Journals (Sweden)

    Hussain Najaf

    2016-01-01

    Full Text Available The design and modeling of Low Stiffness Resilience Shaft (LSRS for the Semi-Active Steering (SAS system using wire ropes is discussed in this paper, along with the static structural torsion test simulation of the wire ropes in order to determine the best possible configuration which serves the purpose of an LSRS. The importance of this study arises due to the unidirectional torsional properties of a wire rope. For an effective operational LSRS, the wire ropes need to have similar angular deflection in both the clockwise and anti-clockwise direction. LSRS, an integral component of the SAS is a flexible shaft that can replace the conventional rigid shaft of the steering system and allows active control to be performed. 3D solid models of the simple strand and the 4 strand wire ropes used in finite element analysis were generated in CAD software SolidWorksTM. The single strand and the different configuration of wire ropes required to function the LSRS effectively were then analyzed using Finite element simulation in ANSYSTM. A single wire rope could not be used because its construction has inconsistency in the torsional stiffness in clockwise and anti-clockwise direction. The single-strand right-direction lay wire rope is found to have 16.05% angular deflection percentage difference in the clockwise and anticlockwise directions which indicates that using a single strand wire rope for the LSRS will cause the vehicle to have a variable response in the clockwise and anti clockwise direction upon turning the steering wheel. Due to this inconsistency, two variations namely Variation 1 and Variation 2 with arrangement of 4 strand wire rope were devised so that the angular deflection percentage difference would be negligible. Simulation results indicated that Variation 1 of the two variations with an angular deflection percentage difference of 0.34% in the clockwise and anti-clockwise direction respectively is best suited for the use in LSRS as it has

  18. Understanding the shape-memory alloys used in orthodontics.

    Science.gov (United States)

    Fernandes, Daniel J; Peres, Rafael V; Mendes, Alvaro M; Elias, Carlos N

    2011-01-01

    Nickel-titanium (NiTi) shape-memory alloys (SMAs) have been used in the manufacture of orthodontic wires due to their shape memory properties, super-elasticity, high ductility, and resistance to corrosion. SMAs have greater strength and lower modulus of elasticity when compared with stainless steel alloys. The pseudoelastic behavior of NiTi wires means that on unloading they return to their original shape by delivering light continuous forces over a wider range of deformation which is claimed to allow dental displacements. The aim of this paper is to discuss the physical, metallurgical, and mechanical properties of NiTi used in Orthodontics in order to analyze the shape memory properties, super-elasticity, and thermomechanical characteristics of SMA.

  19. Assessment of perceived orthodontic appliance attractiveness.

    Science.gov (United States)

    Ziuchkovski, James P; Fields, Henry W; Johnston, William M; Lindsey, Delwin T

    2008-04-01

    To date, no studies have been published in the literature evaluating the esthetics of orthodontic appliances. The purpose of this study was to assess appliance variables, including appliance type, brand, wire, and ligature tie, for their relative attractiveness. Demographic factors were also evaluated. Appliances were placed in an adult, and digital images were captured, standardized, and incorporated into a computer-based survey. Subjects (n = 200) rated each image for attractiveness on a visual analog scale. The attractiveness ratings were analyzed by using factorial analysis of variance with repeated measures. Post-hoc testing was done with the Tukey-Kramer test (overall a appliances), ceramic, and all self-ligating/stainless steel appliances. No differences between brands in each appliance type were found. Ratings of some ceramic appliances were significantly affected by archwire and ligature tie, depending on several demographic interactions. Orthodontic appliance attractiveness varies by the following hierarchy of appliance types: alternative > ceramic > all stainless steel/self-ligating. Wire and tie selection can affect the appearance of ceramic appliances but have no impact on stainless steel or self-ligating appliances. Certain demographic factors have variable effects on attractiveness ratings.

  20. Cells deficient in PARP-1 show an accelerated accumulation of DNA single strand breaks, but not AP sites, over the PARP-1-proficient cells exposed to MMS

    Energy Technology Data Exchange (ETDEWEB)

    Pachkowski, Brian F. [Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, NC (United States); Tano, Keizo [Research Reactor Institute, Kyoto University, Kumatori (Japan); Afonin, Valeriy [Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, NC (United States); Elder, Rhoderick H. [School of Environment and Life Sciences, University of Salford, Greater Manchester (United Kingdom); Takeda, Shunichi [Department of Radiation Genetics, Kyoto University Graduate School of Medicine, Sakyo-ku, Kyoto (Japan); Watanabe, Masami [Research Reactor Institute, Kyoto University, Kumatori (Japan); Swenberg, James A. [Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, NC (United States); Nakamura, Jun, E-mail: ynakamur@email.unc.edu [Department of Environmental Sciences and Engineering, University of North Carolina, Chapel Hill, NC (United States)

    2009-12-01

    Poly(ADP-ribose) polymerase-1 (PARP-1) is a base excision repair (BER) protein that binds to DNA single strand breaks (SSBs) and subsequently synthesizes and transfers poly(ADP-ribose) polymers to various nuclear proteins. Numerous biochemical studies have implicated PARP-1 as a modulator of BER; however, the role of PARP-1 in BER in living cells remains unclear partly due to lack of accurate quantitation of BER intermediates existing in cells. Since DT40 cells, chicken B lymphocytes, naturally lack PARP-2, DT40 cells allow for the investigation of the PARP-1 null phenotype without confounding by PARP-2. To test the hypothesis that PARP-1 is necessary for efficient BER during methylmethane sulfonate (MMS) exposure in vertebrate cells, intact DT40 cells and their isogenic PARP-1 null counterparts were challenged with different exposure scenarios for phenotypic characterization. With chronic exposure, PARP-1 null cells exhibited sensitivity to MMS but with an acute exposure did not accumulate base lesions or AP sites to a greater extent than wild-type cells. However, an increase in SSB content in PARP-1 null cell DNA, as indicated by glyoxal gel electrophoresis under neutral conditions, suggested the presence of BER intermediates. These data suggest that during exposure, PARP-1 impacts the stage of BER after excision of the deoxyribosephosphate moiety from the 5' end of DNA strand breaks by polymerase {beta}.

  1. Direct Binding to Replication Protein A (RPA)-coated Single-stranded DNA Allows Recruitment of the ATR Activator TopBP1 to Sites of DNA Damage*

    Science.gov (United States)

    Acevedo, Julyana; Yan, Shan; Michael, W. Matthew

    2016-01-01

    A critical event for the ability of cells to tolerate DNA damage and replication stress is activation of the ATR kinase. ATR activation is dependent on the BRCT (BRCA1 C terminus) repeat-containing protein TopBP1. Previous work has shown that recruitment of TopBP1 to sites of DNA damage and stalled replication forks is necessary for downstream events in ATR activation; however, the mechanism for this recruitment was not known. Here, we use protein binding assays and functional studies in Xenopus egg extracts to show that TopBP1 makes a direct interaction, via its BRCT2 domain, with RPA-coated single-stranded DNA. We identify a point mutant that abrogates this interaction and show that this mutant fails to accumulate at sites of DNA damage and that the mutant cannot activate ATR. These data thus supply a mechanism for how the critical ATR activator, TopBP1, senses DNA damage and stalled replication forks to initiate assembly of checkpoint signaling complexes. PMID:27129245

  2. Yeast Srs2 Helicase Promotes Redistribution of Single-Stranded DNA-Bound RPA and Rad52 in Homologous Recombination Regulation

    Directory of Open Access Journals (Sweden)

    Luisina De Tullio

    2017-10-01

    Full Text Available Srs2 is a super-family 1 helicase that promotes genome stability by dismantling toxic DNA recombination intermediates. However, the mechanisms by which Srs2 remodels or resolves recombination intermediates remain poorly understood. Here, single-molecule imaging is used to visualize Srs2 in real time as it acts on single-stranded DNA (ssDNA bound by protein factors that function in recombination. We demonstrate that Srs2 is highly processive and translocates rapidly (∼170 nt per second in the 3′→5′ direction along ssDNA saturated with replication protein A (RPA. We show that RPA is evicted from DNA during the passage of Srs2. Remarkably, Srs2 also readily removes the recombination mediator Rad52 from RPA-ssDNA and, in doing so, promotes rapid redistribution of both Rad52 and RPA. These findings have important mechanistic implications for understanding how Srs2 and related nucleic acid motor proteins resolve potentially pathogenic nucleoprotein intermediates.

  3. Yeast Srs2 Helicase Promotes Redistribution of Single-Stranded DNA-Bound RPA and Rad52 in Homologous Recombination Regulation.

    Science.gov (United States)

    De Tullio, Luisina; Kaniecki, Kyle; Kwon, Youngho; Crickard, J Brooks; Sung, Patrick; Greene, Eric C

    2017-10-17

    Srs2 is a super-family 1 helicase that promotes genome stability by dismantling toxic DNA recombination intermediates. However, the mechanisms by which Srs2 remodels or resolves recombination intermediates remain poorly understood. Here, single-molecule imaging is used to visualize Srs2 in real time as it acts on single-stranded DNA (ssDNA) bound by protein factors that function in recombination. We demonstrate that Srs2 is highly processive and translocates rapidly (∼170 nt per second) in the 3'→5' direction along ssDNA saturated with replication protein A (RPA). We show that RPA is evicted from DNA during the passage of Srs2. Remarkably, Srs2 also readily removes the recombination mediator Rad52 from RPA-ssDNA and, in doing so, promotes rapid redistribution of both Rad52 and RPA. These findings have important mechanistic implications for understanding how Srs2 and related nucleic acid motor proteins resolve potentially pathogenic nucleoprotein intermediates. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

  4. Replication stress-induced chromosome breakage is correlated with replication fork progression and is preceded by single-stranded DNA formation.

    Science.gov (United States)

    Feng, Wenyi; Di Rienzi, Sara C; Raghuraman, M K; Brewer, Bonita J

    2011-10-01

    Chromosome breakage as a result of replication stress has been hypothesized to be the direct consequence of defective replication fork progression, or "collapsed" replication forks. However, direct and genome-wide evidence that collapsed replication forks give rise to chromosome breakage is still lacking. Previously we showed that a yeast replication checkpoint mutant mec1-1, after transient exposure to replication impediment imposed by hydroxyurea (HU), failed to complete DNA replication, accumulated single-stranded DNA (ssDNA) at the replication forks, and fragmented its chromosomes. In this study, by following replication fork progression genome-wide via ssDNA detection and by direct mapping of chromosome breakage after HU exposure, we have tested the hypothesis that the chromosome breakage in mec1 cells occurs at collapsed replication forks. We demonstrate that sites of chromosome breakage indeed correlate with replication fork locations. Moreover, ssDNA can be detected prior to chromosome breakage, suggesting that ssDNA accumulation is the common precursor to double strand breaks at collapsed replication forks.

  5. A biomarker model of sublethal genotoxicity (DNA single-strand breaks and adducts) using the sentinel organism Aporrectodea longa in spiked soil

    International Nuclear Information System (INIS)

    Martin, Francis L.; Piearce, Trevor G.; Hewer, Alan; Phillips, David H.; Semple, Kirk T.

    2005-01-01

    There is a need to develop risk biomarkers during the remediation of contaminated land. We employed the earthworm, Aporrectodea longa (Ude), to determine whether genotoxicity measures could be applied to this organism's intestinal tissues. Earthworms were added, for 24 h or 7 days, to soil samples spiked with benzo[a]pyrene (B[a]P) and/or lindane. After exposure, intestinal tissues (crop/gizzard or intestine) were removed prior to the measurement in disaggregated cells of DNA single-strand breaks (SSBs) by the alkaline comet assay. Damage was quantified by comet tail length (CTL, μm). B[a]P 24-h exposure induced dose-related increases (P 32 P-postlabelling, showed a two-adduct-spot pattern. This preliminary investigation suggests that earthworm tissues may be incorporated into genotoxicity assays to facilitate hazard identification within terrestrial ecosystems. - Sublethal genotoxicity in the sentinel organism A. longa can be used to monitor the effects of contaminants in soil

  6. Thermodynamics of complex structures formed between single-stranded DNA oligomers and the KH domains of the far upstream element binding protein

    Energy Technology Data Exchange (ETDEWEB)

    Chakraborty, Kaushik; Sinha, Sudipta Kumar; Bandyopadhyay, Sanjoy, E-mail: sanjoy@chem.iitkgp.ernet.in [Molecular Modeling Laboratory, Department of Chemistry, Indian Institute of Technology, Kharagpur 721302 (India)

    2016-05-28

    The noncovalent interaction between protein and DNA is responsible for regulating the genetic activities in living organisms. The most critical issue in this problem is to understand the underlying driving force for the formation and stability of the complex. To address this issue, we have performed atomistic molecular dynamics simulations of two DNA binding K homology (KH) domains (KH3 and KH4) of the far upstream element binding protein (FBP) complexed with two single-stranded DNA (ss-DNA) oligomers in aqueous media. Attempts have been made to calculate the individual components of the net entropy change for the complexation process by adopting suitable statistical mechanical approaches. Our calculations reveal that translational, rotational, and configurational entropy changes of the protein and the DNA components have unfavourable contributions for this protein-DNA association process and such entropy lost is compensated by the entropy gained due to the release of hydration layer water molecules. The free energy change corresponding to the association process has also been calculated using the Free Energy Perturbation (FEP) method. The free energy gain associated with the KH4–DNA complex formation has been found to be noticeably higher than that involving the formation of the KH3–DNA complex.

  7. Yield of radiation-induced DNA single-strand breaks in Escherichia coli and superinfecting phage lambda at different dose rates. Repair of strand breaks in different buffers

    International Nuclear Information System (INIS)

    Boye, E.; Johansen, I.; Brustad, T.

    1976-01-01

    Cells of E. coli K-12 strain AB 1886 were irradiated in oxygenated phosphate buffered saline at 2 0 C with electrons from a 4-MeV linear accelerator. The yield of DNA single-strand breaks was determined as a function of the dose rate between 2.5 and 21,000 krad/min. For dose rates over 100 krad/min the yield was found to be constant. Below 10 krad/min the yield of breaks decreases drastically. This is explained by rejoining of breaks during irradiation. Twenty percent of the breaks induced by acute exposure are repaired within 3 min at 2 0 C. Superinfecting phage lambda DNA is repaired at the same rate as chromosomal DNA. In contrast to the results obtained with phosphate-buffered saline, an increase in the number of breaks after irradiation is observed when the bacteria are suspended in tris buffer. It is suggested that buffers of low ionic strength facilitate the leakage through the membrane of a small-molecular-weight component(s) necessary for DNA strand rejoining

  8. Lingering single-strand breaks trigger Rad51-independent homology-directed repair of collapsed replication forks in the polynucleotide kinase/phosphatase mutant of fission yeast.

    Directory of Open Access Journals (Sweden)

    Arancha Sanchez

    2017-09-01

    Full Text Available The DNA repair enzyme polynucleotide kinase/phosphatase (PNKP protects genome integrity by restoring ligatable 5'-phosphate and 3'-hydroxyl termini at single-strand breaks (SSBs. In humans, PNKP mutations underlie the neurological disease known as MCSZ, but these individuals are not predisposed for cancer, implying effective alternative repair pathways in dividing cells. Homology-directed repair (HDR of collapsed replication forks was proposed to repair SSBs in PNKP-deficient cells, but the critical HDR protein Rad51 is not required in PNKP-null (pnk1Δ cells of Schizosaccharomyces pombe. Here, we report that pnk1Δ cells have enhanced requirements for Rad3 (ATR/Mec1 and Chk1 checkpoint kinases, and the multi-BRCT domain protein Brc1 that binds phospho-histone H2A (γH2A at damaged replication forks. The viability of pnk1Δ cells depends on Mre11 and Ctp1 (CtIP/Sae2 double-strand break (DSB resection proteins, Rad52 DNA strand annealing protein, Mus81-Eme1 Holliday junction resolvase, and Rqh1 (BLM/WRN/Sgs1 DNA helicase. Coupled with increased sister chromatid recombination and Rad52 repair foci in pnk1Δ cells, these findings indicate that lingering SSBs in pnk1Δ cells trigger Rad51-independent homology-directed repair of collapsed replication forks. From these data, we propose models for HDR-mediated tolerance of persistent SSBs with 3' phosphate in pnk1Δ cells.

  9. Hematopoietic Lineage Transcriptome Stability and Representation in PAXgeneTM Collected Peripheral Blood Utilising SPIA Single-Stranded cDNA Probes for Microarray

    Directory of Open Access Journals (Sweden)

    Laura Kennedy

    2008-01-01

    Full Text Available Peripheral blood as a surrogate tissue for transcriptome profiling holds great promise for the discovery of diagnostic and prognostic disease biomarkers, particularly when target tissues of disease are not readily available. To maximize the reliability of gene expression data generated from clinical blood samples, both the sample collection and the microarray probe generation methods should be optimized to provide stabilized, reproducible and representative gene expression profiles faithfully representing the transcriptional profiles of the constituent blood cell types present in the circulation. Given the increasing innovation in this field in recent years, we investigated a combination of methodological advances in both RNA stabilisation and microarray probe generation with the goal of achieving robust, reliable and representative transcriptional profiles from whole blood. To assess the whole blood profiles, the transcriptomes of purified blood cell types were measured and compared with the global transcriptomes measured in whole blood. The results demonstrate that a combination of PAXgeneTM RNA stabilising technology and single-stranded cDNA probe generation afforded by the NuGEN Ovation RNA amplification system V2TM enables an approach that yields faithful representation of specific hematopoietic cell lineage transcriptomes in whole blood without the necessity for prior sample fractionation, cell enrichment or globin reduction. Storage stability assessments of the PAXgeneTM blood samples also advocate a short, fixed room temperature storage time for all PAXgeneTM blood samples collected for the purposes of global transcriptional profiling in clinical studies.

  10. Hematopoietic Lineage Transcriptome Stability and Representation in PAXgene Collected Peripheral Blood Utilising SPIA Single-Stranded cDNA Probes for Microarray.

    Science.gov (United States)

    Kennedy, Laura; Vass, J Keith; Haggart, D Ross; Moore, Steve; Burczynski, Michael E; Crowther, Dan; Miele, Gino

    2008-08-25

    Peripheral blood as a surrogate tissue for transcriptome profiling holds great promise for the discovery of diagnostic and prognostic disease biomarkers, particularly when target tissues of disease are not readily available. To maximize the reliability of gene expression data generated from clinical blood samples, both the sample collection and the microarray probe generation methods should be optimized to provide stabilized, reproducible and representative gene expression profiles faithfully representing the transcriptional profiles of the constituent blood cell types present in the circulation. Given the increasing innovation in this field in recent years, we investigated a combination of methodological advances in both RNA stabilisation and microarray probe generation with the goal of achieving robust, reliable and representative transcriptional profiles from whole blood. To assess the whole blood profiles, the transcriptomes of purified blood cell types were measured and compared with the global transcriptomes measured in whole blood. The results demonstrate that a combination of PAXgene() RNA stabilising technology and single-stranded cDNA probe generation afforded by the NuGEN Ovation RNA amplification system V2() enables an approach that yields faithful representation of specific hematopoietic cell lineage transcriptomes in whole blood without the necessity for prior sample fractionation, cell enrichment or globin reduction. Storage stability assessments of the PAXgene() blood samples also advocate a short, fixed room temperature storage time for all PAXgene() blood samples collected for the purposes of global transcriptional profiling in clinical studies.

  11. Hematopoietic Lineage Transcriptome Stability and Representation in PAXgene™ Collected Peripheral Blood Utilising SPIA Single-Stranded cDNA Probes for Microarray

    Science.gov (United States)

    Kennedy, Laura; Vass, J. Keith; Haggart, D. Ross; Moore, Steve; Burczynski, Michael E.; Crowther, Dan; Miele, Gino

    2008-01-01

    Peripheral blood as a surrogate tissue for transcriptome profiling holds great promise for the discovery of diagnostic and prognostic disease biomarkers, particularly when target tissues of disease are not readily available. To maximize the reliability of gene expression data generated from clinical blood samples, both the sample collection and the microarray probe generation methods should be optimized to provide stabilized, reproducible and representative gene expression profiles faithfully representing the transcriptional profiles of the constituent blood cell types present in the circulation. Given the increasing innovation in this field in recent years, we investigated a combination of methodological advances in both RNA stabilisation and microarray probe generation with the goal of achieving robust, reliable and representative transcriptional profiles from whole blood. To assess the whole blood profiles, the transcriptomes of purified blood cell types were measured and compared with the global transcriptomes measured in whole blood. The results demonstrate that a combination of PAXgene™ RNA stabilising technology and single-stranded cDNA probe generation afforded by the NuGEN Ovation RNA amplification system V2™ enables an approach that yields faithful representation of specific hematopoietic cell lineage transcriptomes in whole blood without the necessity for prior sample fractionation, cell enrichment or globin reduction. Storage stability assessments of the PAXgene™ blood samples also advocate a short, fixed room temperature storage time for all PAXgene™ blood samples collected for the purposes of global transcriptional profiling in clinical studies. PMID:19578521

  12. A fluorescence-based polymerase chain reaction-linked single-strand conformation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp.

    Science.gov (United States)

    Alasaad, Samer; Soriguer, Ramón C; Abu-Madi, Marawan; El Behairy, Ahmed; Baños, Pablo Díez; Píriz, Ana; Fickel, Joerns; Zhu, Xing-Quan

    2011-06-01

    The present study aimed to establish a fluorescence-based polymerase chain reaction-linked single-strand conformation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp. Based on the sequences of the second internal transcribed spacer (ITS-2) of the nuclear ribosomal DNA, we designed a set of genus-specific primers for the amplification of Fasciola ITS-2, with an estimated size of 140 bp. These primers were labelled by fluorescence dyes, and the PCR products were analyzed by capillary electrophoresis under non-denaturing conditions (F-PCR-SSCP). Capillary electrophoresis analysis of the fluorescence-labelled DNA fragments displayed three different peak profiles that allowed the accurate identification of Fasciola species: one single peak specific for either Fasciola hepatica or Fasciola gigantica and a doublet peak corresponding to the "intermediate" Fasciola. Validation of our novel method was performed using Fasciola specimens from different host animals from China, Spain, Nigeria, and Egypt. This F-PCR-SSCP assay provides a rapid, simple, and robust tool for the identification and differentiation between Fasciola spp.

  13. Analysis of Coinfections with A/H1N1 Strain Variants among Pigs in Poland by Multitemperature Single-Strand Conformational Polymorphism

    Directory of Open Access Journals (Sweden)

    Krzysztof Lepek

    2015-01-01

    Full Text Available Monitoring and control of infections are key parts of surveillance systems and epidemiological risk prevention. In the case of influenza A viruses (IAVs, which show high variability, a wide range of hosts, and a potential of reassortment between different strains, it is essential to study not only people, but also animals living in the immediate surroundings. If understated, the animals might become a source of newly formed infectious strains with a pandemic potential. Special attention should be focused on pigs, because of the receptors specific for virus strains originating from different species, localized in their respiratory tract. Pigs are prone to mixed infections and may constitute a reservoir of potentially dangerous IAV strains resulting from genetic reassortment. It has been reported that a quadruple reassortant, A(H1N1pdm09, can be easily transmitted from humans to pigs and serve as a donor of genetic segments for new strains capable of infecting humans. Therefore, it is highly desirable to develop a simple, cost-effective, and rapid method for evaluation of IAV genetic variability. We describe a method based on multitemperature single-strand conformational polymorphism (MSSCP, using a fragment of the hemagglutinin (HA gene, for detection of coinfections and differentiation of genetic variants of the virus, difficult to identify by conventional diagnostic.

  14. Combining Single Strand Oligodeoxynucleotides and CRISPR/Cas9 to Correct Gene Mutations in β-Thalassemia-induced Pluripotent Stem Cells.

    Science.gov (United States)

    Niu, Xiaohua; He, Wenyin; Song, Bing; Ou, Zhanhui; Fan, Di; Chen, Yuchang; Fan, Yong; Sun, Xiaofang

    2016-08-05

    β-Thalassemia (β-Thal) is one of the most common genetic diseases in the world. The generation of patient-specific β-Thal-induced pluripotent stem cells (iPSCs), correction of the disease-causing mutations in those cells, and then differentiation into hematopoietic stem cells offers a new therapeutic strategy for this disease. Here, we designed a CRISPR/Cas9 to specifically target the Homo sapiens hemoglobin β (HBB) gene CD41/42(-CTTT) mutation. We demonstrated that the combination of single strand oligodeoxynucleotides with CRISPR/Cas9 was capable of correcting the HBB gene CD41/42 mutation in β-Thal iPSCs. After applying a correction-specific PCR assay to purify the corrected clones followed by sequencing to confirm mutation correction, we verified that the purified clones retained full pluripotency and exhibited normal karyotyping. Additionally, whole-exome sequencing showed that the mutation load to the exomes was minimal after CRISPR/Cas9 targeting. Furthermore, the corrected iPSCs were selected for erythroblast differentiation and restored the expression of HBB protein compared with the parental iPSCs. This method provides an efficient and safe strategy to correct the HBB gene mutation in β-Thal iPSCs. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  15. Cells deficient in PARP-1 show an accelerated accumulation of DNA single strand breaks, but not AP sites, over the PARP-1-proficient cells exposed to MMS.

    Science.gov (United States)

    Pachkowski, Brian F; Tano, Keizo; Afonin, Valeriy; Elder, Rhoderick H; Takeda, Shunichi; Watanabe, Masami; Swenberg, James A; Nakamura, Jun

    2009-12-01

    Poly(ADP-ribose) polymerase-1 (PARP-1) is a base excision repair (BER) protein that binds to DNA single strand breaks (SSBs) and subsequently synthesizes and transfers poly(ADP-ribose) polymers to various nuclear proteins. Numerous biochemical studies have implicated PARP-1 as a modulator of BER; however, the role of PARP-1 in BER in living cells remains unclear partly due to lack of accurate quantitation of BER intermediates existing in cells. Since DT40 cells, chicken B lymphocytes, naturally lack PARP-2, DT40 cells allow for the investigation of the PARP-1 null phenotype without confounding by PARP-2. To test the hypothesis that PARP-1 is necessary for efficient BER during methylmethane sulfonate (MMS) exposure in vertebrate cells, intact DT40 cells and their isogenic PARP-1 null counterparts were challenged with different exposure scenarios for phenotypic characterization. With chronic exposure, PARP-1 null cells exhibited sensitivity to MMS but with an acute exposure did not accumulate base lesions or AP sites to a greater extent than wild-type cells. However, an increase in SSB content in PARP-1 null cell DNA, as indicated by glyoxal gel electrophoresis under neutral conditions, suggested the presence of BER intermediates. These data suggest that during exposure, PARP-1 impacts the stage of BER after excision of the deoxyribosephosphate moiety from the 5' end of DNA strand breaks by polymerase beta.

  16. Cas3 is a single-stranded DNA nuclease and ATP-dependent helicase in the CRISPR/Cas immune system.

    Science.gov (United States)

    Sinkunas, Tomas; Gasiunas, Giedrius; Fremaux, Christophe; Barrangou, Rodolphe; Horvath, Philippe; Siksnys, Virginijus

    2011-04-06

    Clustered regularly interspaced short palindromic repeat (CRISPR) is a recently discovered adaptive prokaryotic immune system that provides acquired immunity against foreign nucleic acids by utilizing small guide crRNAs (CRISPR RNAs) to interfere with invading viruses and plasmids. In Escherichia coli, Cas3 is essential for crRNA-guided interference with virus proliferation. Cas3 contains N-terminal HD phosphohydrolase and C-terminal Superfamily 2 (SF2) helicase domains. Here, we provide the first report of the cloning, expression, purification and in vitro functional analysis of the Cas3 protein of the Streptococcus thermophilus CRISPR4 (Ecoli subtype) system. Cas3 possesses a single-stranded DNA (ssDNA)-stimulated ATPase activity, which is coupled to unwinding of DNA/DNA and RNA/DNA duplexes. Cas3 also shows ATP-independent nuclease activity located in the HD domain with a preference for ssDNA substrates. To dissect the contribution of individual domains, Cas3 separation-of-function mutants (ATPase(+)/nuclease(-) and ATPase(-)/nuclease(+)) were obtained by site-directed mutagenesis. We propose that the Cas3 ATPase/helicase domain acts as a motor protein, which assists delivery of the nuclease activity to Cascade-crRNA complex targeting foreign DNA.

  17. Chemical shift changes provide evidence for overlapping single-stranded DNA and XPA binding sites on the 70 kDa subunit of human replication protein A

    Energy Technology Data Exchange (ETDEWEB)

    Daughdrill, Gary W.; Buchko, Garry W.; Botuyan, Maria V.; Arrowsmith, Cheryl H.; Wold, Marc S.; Kennedy, Michael A.; Lowry, David F.

    2003-07-15

    Replication protein A (RPA) is a heterotrimeric single-stranded DNA (ssDNA) binding protein that can form a complex with the xeroderma pigmentosum group A protein (XPA). This complex can preferentially recognize UV damaged DNA over undamaged DNA and has been implicated in the stabilization of open complex formation during nucleotide excision repair. In this report, NMR spectroscopy was used to investigate the interaction between a fragment of the 70 kDa subunit of human RPA, residues 1-326 (hRPA701-326), and a fragment of the human XPA protein, residues 98-219 (XPA-MBD). Intensity changes were observed for amide resonances in the 1H-15N correlation spectrum of uniformly 15N-labeled hRPA701-326 after the addition of unlabeled XPA-MBD. The intensity changes observed were restricted to an ssDNA binding domain that is between residues 183 and 296 of the hRPA701-326 fragment. The hRPA701-326 residues with the largest resonance intensity reductions were mapped onto the structure of the ssDNA binding domain to identify the binding surface with XPA-MBD. The XPA-MBD binding surface showed significant overlap with an ssDNA binding surface that was previously identified using NMR spectroscopy and X-ray crystallography.

  18. The influence of inhibitors on dimer removal and repair of single-strand breaks in normal and bromodeoxyuridine substituted DNA of HeLa cells

    International Nuclear Information System (INIS)

    Cornelis, J.J.

    1978-01-01

    The elimination of cyclobutane pyrimidine dimers from the nuclear DNA of ultraviolet irradiated HeLa cells has been examined by means of chromatography and immunoautoradiography. The extent and duration of the process was similar when dimers were assayed by both methods, proving that the antisera recognized pyrimidine dimers. The rate of dimer excision did not differ through the cell cycle with the exception of mitosis during which no dimers were removed. Dimer excision is a relatively fast process which is terminated within a few hours, but it leaves many dimers in the DNA. Excision is depressed by inhibitors of semiconservative DNA synthesis that affect the DNA precursor pool or DNA polymerases. Cells whose DNA is partly substituted with bromodeoxyuridine instead of thymidine, repair single-strand breaks and remove dimers at the same rate but to different extents. On the other hand, inhibitors limit repair of breaks and removal of dimers to the same degree suggesting that the repair of the two types of lesion is coordinated. (Auth.)

  19. Single-strand breaks in the DNA of the uvrA and uvrB strains of Escherichia coli K-12 after ultraviolet irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Youngs, D A; Smith, K C [Stanford Univ., Calif. (USA). Dept. of Radiology

    1976-12-01

    DNA single-strand breaks were produced in uvrA and uvrB strains of E.coli K-12 after UV (254 nm) irradiation. These breaks appeared to be produced both directly by photochemical events, and by a temperature-dependent process. Cyclobutane-type pyrimidine dimers are probably not the photoproducts that lead to the temperature-dependent breaks, since photoreactivation had no detectable effect on the final yield of breaks. The DNA strand breaks appeared to be repairable by a process that requires DNA polymerase I and polynucleotide ligase, but not the recA, recB, recF, lexA101 or uvrD gene products. It is hypothesized that these temperature-dependent breaks occur either as a result of breakdown of a thermolabile photoproduct, or as the initial endonucleolytic event of a uvrA, uvrB-independent excision repair process that acts on a UV photoproduct other than the cyclobutane-type pyrimidine dimer.

  20. Molecular mechanisms of induced mutagenesis. Replication in vivo of bacteriophage phiX174 single-stranded, ultraviolet light-irradiated DNA in intact and irradiated host cells

    Energy Technology Data Exchange (ETDEWEB)

    Caillet-Fauquet, P; Defais, M; Radman, M [Brussels Univ. (Belgium)

    1977-11-25

    Genetic analysis has revealed that radiation and many chemical mutagens induce in bacteria an error-prone DNA repair process which is responsible for their mutagenic effect. The biochemical mechanism of this inducible error-prone repair has been studied by analysis of the first round of DNA synthesis on ultraviolet light-irradiated phiX174 DNA in both intact and ultraviolet light-irradiated host cells. Intracellular phiX174 DNA was extracted, subjected to isopycnic CsCl density-gradient analysis, hydroxylapatite chromatography and digestion by single-strand-specific endonuclease S/sub 1/. Ultraviolet light-induced photolesions in viral DNA cause a permanent blockage of DNA synthesis in intact Escherichia coli cells. However, when host cells were irradiated and incubated to induce fully the error-prone repair system, a significant fraction of irradiated phiX174 DNA molecules can be fully replicated. Thus, inducible error-prone repair in E.coli is manifested by an increased capacity for DNA synthesis on damaged phiX174 DNA. Chloramphenicol (100 ..mu.. g/ml), which is an inhibitor of the inducible error-prone DNA repair, is also an inhibitor of this particular inducible DNA synthesis.

  1. In Vitro Selection of Single-Stranded DNA Molecular Recognition Elements against S. aureus Alpha Toxin and Sensitive Detection in Human Serum

    Directory of Open Access Journals (Sweden)

    Ka L. Hong

    2015-01-01

    Full Text Available Alpha toxin is one of the major virulence factors secreted by Staphylococcus aureus, a bacterium that is responsible for a wide variety of infections in both community and hospital settings. Due to the prevalence of S. aureus related infections and the emergence of methicillin-resistant S. aureus, rapid and accurate diagnosis of S. aureus infections is crucial in benefiting patient health outcomes. In this study, a rigorous Systematic Evolution of Ligands by Exponential Enrichment (SELEX variant previously developed by our laboratory was utilized to select a single-stranded DNA molecular recognition element (MRE targeting alpha toxin with high affinity and specificity. At the end of the 12-round selection, the selected MRE had an equilibrium dissociation constant (Kd of 93.7 ± 7.0 nM. Additionally, a modified sandwich enzyme-linked immunosorbent assay (ELISA was developed by using the selected ssDNA MRE as the toxin-capturing element and a sensitive detection of 200 nM alpha toxin in undiluted human serum samples was achieved.

  2. Fluoride influences nickel-titanium orthodontic wires′ surface texture and friction resistance

    Directory of Open Access Journals (Sweden)

    Mona Aly Abbassy

    2016-01-01

    Full Text Available Objectives: The aim of this study was to investigate the effects exerted by the acidulated fluoride gel on stainless steel and nickel-titanium (Ni-Ti orthodontic wires. Materials and Methods: Sixty stainless steel and Ni-Ti orthodontic archwires were distributed into forty archwires used for in vitro study and twenty for in situ study. Fluoride was applied for 1 h in the in vitro experiment while it was applied for 5 min in the in situ experiment. The friction resistance of all wires with ceramic brackets before/after topical fluoride application was measured using a universal testing machine at 1 min intervals of moving wire. Moreover, surface properties of the tested wires before/after fluoride application and before/after friction test were examined by a scanning electron microscope (SEM. Dunnett′s t-test was used to compare frictional resistance of as-received stainless steel wires and Ni-Ti wires to the wires treated by fluoride in vitro and in situ (P < 0.05. Two-way ANOVA was used to compare the effect of fluoride application and type of wire on friction resistance in vitro and in situ (P < 0.05. Results: Ni-Ti wires recorded significantly high friction resistance after fluoride application when compared to stainless steel wires in vitro, P < 0.05. Fluoride application did not significantly affect the friction resistance of the tested wires in situ, P < 0.05. SEM observation revealed deterioration of the surface texture of the Ni-Ti wires after fluoride application in vitro and in situ. Conclusions: The in vitro fluoride application caused an increase in friction resistance of Ni-Ti wires when compared to stainless steel wires. In vitro and in situ fluoride application caused deterioration in surface properties of Ni-Ti wires.

  3. [Evaluation of orthodontic friction using a tribometer with alternating movement].

    Science.gov (United States)

    Pernier, C M; Jablonska-Mazanek, E D; Ponsonnet, L; Grosgogeat, B

    2005-12-01

    It is essential for orthodontists to control the complex phenomenon of friction. The in vitro techniques, usually dynamometers or tensile testing machines, used to measure the frictional resistance between arch wires and brackets are linear and unidirectional and can be criticised because tooth movements, such as tipping and uprighting, as well everyday oral activities, primarily chewing, are not uni-dimensional but more closely resemble the small amplitude oscillatory phenomena known as fretting. We therefore decided to develop a fretting machine not with linear but with alternating movements better suited to evaluate the frictional behaviour of orthodontic bracket-wire combinations. Once we had completed construction of this device, we proceeded to measure the frictional resistance between one stainless steel bracket (MicroArch GAC) and five wires currently used in orthodontics (Two nickel-titanium shape memory alloys: Neo Sentalloy and Neo Sentalloy with Ionguard GAC--Three titanium-molybdenum alloys: TMA and Low Friction TMA Ormco and Resolve GAC). We were able to set up a classification of the wires according to their coefficient of friction, demonstrating the inefficacy of ion implantation and quantifying the increase in the coefficient of friction which occurs when Resolve wires are placed in the oral environment for approximately one year.

  4. Digital workflows in contemporary orthodontics

    Directory of Open Access Journals (Sweden)

    Lars R Christensen

    2017-01-01

    Full Text Available Digital workflows are now increasingly possible in orthodontic practice. Workflows designed to improve the customization of orthodontic appliances are now available through laboratories and orthodontic manufacturing facilities in many parts of the world. These now have the potential to improve certain aspects of patient care.

  5. Copper, Aluminum and Nickel: A New Monocrystalline Orthodontic Alloy

    Science.gov (United States)

    Wierenga, Mark

    Introduction: This study was designed to evaluate, via tensile and bend testing, the mechanical properties of a newly-developed monocrystalline orthodontic archwire comprised of a blend of copper, aluminum, and nickel (CuAlNi). Methods: The sample was comprised of three shape memory alloys; CuAlNi, copper nickel titanium (CuNiTi), and nickel titanium (NiTi); from various orthodontic manufacturers in both 0.018" round and 0.019" x 0.025" rectangular dimensions. Additional data was gathered for similarly sized stainless steel and beta-titanium archwires as a point of reference for drawing conclusions about the relative properties of the archwires. Measurements of loading and unloading forces were recorded in both tension and deflection testing. Repeated-measure ANOVA (alpha= 0.05) was used to compare loading and unloading forces across wires and one-way ANOVA (alpha= 0.05) was used to compare elastic moduli and hysteresis. To identify significant differences, Tukey post-hoc comparisons were performed. Results: The modulus of elasticity, deflection forces, and hysteresis profiles of CuAlNi were significantly different than the other superelastic wires tested. In all tests, CuAlNi had a statistically significant lower modulus of elasticity compared to the CuNiTi and NiTi wires (P orthodontic metallurgy.

  6. Pacemaker wires

    International Nuclear Information System (INIS)

    Fransson, S.G.

    1993-01-01

    Evaluation of pacemaker wires were performed by comparing Advanced Multiple Beam Equalization Radiography (AMBER) with conventional chest radiography. The scanning equalization technique of the AMBER unit makes it superior to conventional technique in the depiction of different structures in the mediastinum or in the pleural sinuses. So far motion artifacts have not been considered clinically important. The longer exposure time, however, may impair the assessment of pacemaker wires. The motion artifact described may not only make adequate evaluation impossible but may even give a false impression of a lead fracture. The difference between the two systems was significant. (orig.)

  7. Two modes of interaction of the single-stranded DNA-binding protein of bacteriophage T7 with the DNA polymerase-thioredoxin complex

    KAUST Repository

    Ghosh, Sharmistha; Hamdan, Samir; Richardson, Charles C.

    2010-01-01

    The DNA polymerase encoded by bacteriophage T7 has low processivity. Escherichia coli thioredoxin binds to a segment of 76 residues in the thumb subdomain of the polymerase and increases the processivity. The binding of thioredoxin leads to the formation of two basic loops, loops A and B, located within the thioredoxin-binding domain (TBD). Both loops interact with the acidic C terminus of the T7 helicase. A relatively weak electrostatic mode involves the C-terminal tail of the helicase and the TBD, whereas a high affinity interaction that does not involve the C-terminal tail occurs when the polymerase is in a polymerization mode. T7 gene 2.5 single-stranded DNA-binding protein (gp2.5) also has an acidic C-terminal tail. gp2.5 also has two modes of interaction with the polymerase, but both involve the C-terminal tail of gp2.5. An electrostatic interaction requires the basic residues in loops A and B, and gp2.5 binds to both loops with similar affinity as measured by surface plasmon resonance. When the polymerase is in a polymerization mode, the C terminus of gene 2.5 protein interacts with the polymerase in regions outside the TBD.gp2.5 increases the processivity of the polymerase-helicase complex during leading strand synthesis. When loop B of the TBD is altered, abortive DNA products are observed during leading strand synthesis. Loop B appears to play an important role in communication with the helicase and gp2.5, whereas loop A plays a stabilizing role in these interactions. © 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

  8. Effect of vanillin on methylene blue plus light-induced single-strand breaks in plasmid pBR322 DNA.

    Science.gov (United States)

    Kumar, S S; Ghosh, A; Devasagayam, T P; Chauhan, P S

    2000-09-20

    The ability of vanillin (4-hydroxy-3-methoxybenzaldehyde), a naturally occurring food flavouring agent, in inhibiting photosensitization-induced single-strand breaks (ssbs) in plasmid pBR322 DNA has been examined in an in vitro system, independent of DNA repair/replication processes. Photosensitization of DNA with methylene blue, visible light and oxygen, induced ssbs resulting in the production of open circular form (OC form) in a concentration-dependent manner. The yield of OC form induced by photosensitization was increased several-fold by deuteration of the buffer and was found to be inhibited by sodium azide, a scavenger of singlet oxygen (1O(2)). Vanillin, per se, did not induce but inhibited photosensitization-induced ssbs in plasmid DNA, at millimolar concentrations. The inhibitory effect of vanillin was both concentration- and time-dependent. On a molar basis, vanillin was, however, less effective than trolox, a water-soluble analogue of alpha-tocopherol. Photosensitization by methylene blue system generates singlet oxygen, as one of the major components of ROS. Therefore, interaction of singlet oxygen with vanillin was investigated. The rate constant of vanillin with 1O(2) was estimated to be 5.93x10(7)M(-1)s(-1) and that of sodium azide as 2. 7x10(8)M(-1)s(-1). The present investigations show that vanillin can protect against photosensitization-induced ssbs in the plasmid pBR322 DNA, and this effect may partly be due to its ability to scavenge 1O(2).

  9. Gauging the Nanotoxicity of h2D-C2N toward Single-Stranded DNA: An in Silico Molecular Simulation Approach.

    Science.gov (United States)

    Mukhopadhyay, Titas Kumar; Bhattacharyya, Kalishankar; Datta, Ayan

    2018-04-12

    Recent toxicological assessments of graphene, graphene oxides, and some other two-dimensional (2D) materials have shown them to be substantially toxic at the nanoscale, where they inhibit and eventually disrupt biological processes. These shortfalls of graphene and analogs have resulted in a quest for novel biocompatible 2D materials with minimum cytotoxicity. In this article, we demonstrate C 2 N (h2D-C 2 N), a newly synthesized 2D porous graphene analog, to be non-nanotoxic toward genetic materials from an "in-silico" point of view through sequence-dependent binding of different polynucleotide single-stranded DNA (ssDNA) onto it. The calculated binding energy of nucleobases and the free energy of binding of polynucleotides follow the common trait, cytosine > guanine > adenine > thymine, and are well within the limits of physisorption. Ab-initio simulations completely exclude the possibility of any chemical reaction, demonstrating purely noncovalent binding of nucleobases with C 2 N through a crucial interplay between hydrogen bonding and π-stacking interactions with the surface. Further, we show that the extent of distortion inflicted upon ssDNA by C 2 N is negligible. Analysis of the density of states of the nucleobase-C 2 N hybrids confirms minimum electronic perturbation of the bases after adsorption. Most importantly, we demonstrate the potency of C 2 N in nucleic acid transportation via reversible binding of ssDNA. The plausible use of C 2 N as a template for DNA repair is illustrated through an example of C 2 N-assisted complementary ssDNA winding.

  10. Variabilidad genética de Aedes aegypti en algunas áreas del Perú usando Single Stranded Conformational Polymorphism (SSCP

    Directory of Open Access Journals (Sweden)

    Nélida Leiva G

    2004-07-01

    Full Text Available Aedes aegypti es el vector responsable de la transmisión del virus del dengue, su distribución geográfica se ha ampliado rápidamente debido principalmente a la intervención de los seres humanos. Objetivo: Analizar la variabilidad genética de este mosquito mediante la comparación del Segundo Espaciador Transcrito Interno (ITS 2 perteneciente al ADN ribosomal (rADN. Materiales y Métodos: Se analizaron muestras de ocho localidades (Jaén, Tingo María, Iquitos, Lambayeque, el distrito de El Rimac, Sullana y Zarumilla y uno de la provincia de Huaquillas (Ecuador. El análisis de la variabilidad se determinó usando la técnica conocida como SSCP (Single Stranded Conformation Polymorphism. Resultados: El estudio muestra que existe variabilidad genética entre las poblaciones analizadas, principalmente entre las muestras localizadas en la costa del Perú (Zarumilla, El Rímac, Sullana y Huaquillas y las muestras del nororiente (Tingo María, Iquitos, Jaén y Lambayeque Conclusión: Se determinaron dos variantes genéticas entre las poblaciones de Aedes aegypti: Costeña y Nororiental, que probablemente provienen de dos ancestros diferentes y cuyo ancestro común sufrió de aislamiento por distancia. Se observó que no existe relación entre las distancias genéticas y las distancias geográficas indicando que la migración de estas poblaciones es el resultado de la intervención de los seres humanos que diseminan al vector y no por la migración activa del mosquito. Se plantea el papel de la Cordillera de los Andes en la migración y separación de las poblaciones de Aedes.

  11. Single-stranded DNA aptamer targeting and neutralization of anti-D alloantibody: a potential therapeutic strategy for haemolytic diseases caused by Rhesus alloantibody.

    Science.gov (United States)

    Zhang, Yinze; Wu, Fan; Wang, Manni; Zhuang, Naibao; Zhou, Huayou; Xu, Hua

    2018-02-01

    Rhesus (Rh) D antigen is the most important antigen in the Rh blood group system because of its strong immunogenicity. When RhD-negative individuals are exposed to RhD-positive blood, they may produce anti-D alloantibody, potentially resulting in delayed haemolytic transfusion reactions and Rh haemolytic disease of the foetus and newborn, which are difficult to treat. Inhibition of the binding of anti-D antibody with RhD antigens on the surface of red blood cells may effectively prevent immune haemolytic diseases. In this study, single-stranded (ss) DNA aptamers, specifically binding to anti-D antibodies, were selected via systematic evolution of ligands by exponential enrichment (SELEX) technology. After 14 rounds of selection, the purified ssDNA was sequenced using a Personal Genome Machine system. Haemagglutination inhibition assays were performed to screen aptamers for biological activity in terms of blocking antigen-antibody reactions: the affinity and specificity of the aptamers were also determined. In addition to high specificity, the aptamers which were selected showed high affinity for anti-D antibodies with dissociation constant (K d ) values ranging from 51.46±14.90 to 543.30±92.59 nM. By the combined use of specific ssDNA aptamer 7 and auxiliary ssDNA aptamer 2, anti-D could be effectively neutralised at low concentrations of the aptamers. Our results demonstrate that ssDNA aptamers may be a novel, promising strategy for the treatment of delayed haemolytic transfusion reactions and Rh haemolytic disease of the foetus and newborn.

  12. Two modes of interaction of the single-stranded DNA-binding protein of bacteriophage T7 with the DNA polymerase-thioredoxin complex

    KAUST Repository

    Ghosh, Sharmistha

    2010-04-06

    The DNA polymerase encoded by bacteriophage T7 has low processivity. Escherichia coli thioredoxin binds to a segment of 76 residues in the thumb subdomain of the polymerase and increases the processivity. The binding of thioredoxin leads to the formation of two basic loops, loops A and B, located within the thioredoxin-binding domain (TBD). Both loops interact with the acidic C terminus of the T7 helicase. A relatively weak electrostatic mode involves the C-terminal tail of the helicase and the TBD, whereas a high affinity interaction that does not involve the C-terminal tail occurs when the polymerase is in a polymerization mode. T7 gene 2.5 single-stranded DNA-binding protein (gp2.5) also has an acidic C-terminal tail. gp2.5 also has two modes of interaction with the polymerase, but both involve the C-terminal tail of gp2.5. An electrostatic interaction requires the basic residues in loops A and B, and gp2.5 binds to both loops with similar affinity as measured by surface plasmon resonance. When the polymerase is in a polymerization mode, the C terminus of gene 2.5 protein interacts with the polymerase in regions outside the TBD.gp2.5 increases the processivity of the polymerase-helicase complex during leading strand synthesis. When loop B of the TBD is altered, abortive DNA products are observed during leading strand synthesis. Loop B appears to play an important role in communication with the helicase and gp2.5, whereas loop A plays a stabilizing role in these interactions. © 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

  13. Novel Positive-Sense, Single-Stranded RNA (+ssRNA) Virus with Di-Cistronic Genome from Intestinal Content of Freshwater Carp (Cyprinus carpio)

    Science.gov (United States)

    Pankovics, Péter; Simmonds, Peter

    2011-01-01

    A novel positive-sense, single-stranded RNA (+ssRNA) virus (Halastavi árva RNA virus, HalV; JN000306) with di-cistronic genome organization was serendipitously identified in intestinal contents of freshwater carps (Cyprinus carpio) fished by line-fishing from fishpond “Lőrinte halastó” located in Veszprém County, Hungary. The complete nucleotide (nt) sequence of the genomic RNA is 9565 nt in length and contains two long - non-in-frame - open reading frames (ORFs), which are separated by an intergenic region. The ORF1 (replicase) is preceded by an untranslated sequence of 827 nt, while an untranslated region of 139 nt follows the ORF2 (capsid proteins). The deduced amino acid (aa) sequences of the ORFs showed only low (less than 32%) and partial similarity to the non-structural (2C-like helicase, 3C-like cystein protease and 3D-like RNA dependent RNA polymerase) and structural proteins (VP2/VP4/VP3) of virus families in Picornavirales especially to members of the viruses with dicistronic genome. Halastavi árva RNA virus is present in intestinal contents of omnivorous freshwater carps but the origin and the host species of this virus remains unknown. The unique viral sequence and the actual position indicate that Halastavi árva RNA virus seems to be the first member of a new di-cistronic ssRNA virus. Further studies are required to investigate the specific host species (and spectrum), ecology and role of Halastavi árva RNA virus in the nature. PMID:22195010

  14. Detection of rifampin resistance patterns in Mycobacterium tuberculosis strains isolated in Iran by polymerase chain reaction-single-strand conformation polymorphism and direct sequencing methods

    Directory of Open Access Journals (Sweden)

    Bahram Nasr Isfahani

    2006-09-01

    Full Text Available Mutations in the rpoB locus confer conformational changes leading to defective binding of rifampin (RIF to rpoB and consequently resistance in Mycobacterium tuberculosis. Polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP was established as a rapid screening test for the detection of mutations in the rpoB gene, and direct sequencing has been unambiguously applied to characterize mutations. A total of 37 of Iranian isolates of M. tuberculosis, 16 sensitive and 21 resistant to RIF, were used in this study. A 193-bp region of the rpoB gene was amplified and PCR-SSCP patterns were determined by electrophoresis in 10% acrylamide gel and silver staining. Also, 21 samples of 193-bp rpoB amplicons with different PCR-SSCP patterns from RIFr and 10 from RIFs were sequenced. Seven distinguishable PCR-SSCP patterns were recognized in the 21 Iranian RIFr strains, while 15 out of 16 RIFs isolates demonstrated PCR-SSCP banding patterns similar to that of sensitive standard strain H37Rv. However one of the sensitive isolates demonstrated a different pattern. There were seen six different mutations in the amplified region of rpoB gene: codon 516(GAC/GTC, 523(GGG/GGT, 526(CAC/TAC, 531(TCG/TTG, 511(CTG/TTG, and 512(AGC/TCG. This study demonstrated the high specificity (93.8% and sensitivity (95.2% of PCR-SSCP method for detection of mutation in rpoB gene; 85.7% of RIFr strains showed a single mutation and 14.3% had no mutations. Three strains showed mutations caused polymorphism. Our data support the common notion that rifampin resistance genotypes are generally present mutations in codons 531 and 526, most frequently found in M. tuberculosis populations regardless of geographic origin.

  15. Removable orthodontic appliance with nickel-titanium spring to reposition the upper incisors in an autistic patient.

    Science.gov (United States)

    Saito, Kan; Jang, Insan; Kubota, Kazumi; Hoshino, Tomonori; Hotokezaka, Hitoshi; Yoshida, Noriaki; Fujiwara, Taku

    2013-01-01

    A newly designed removable appliance with a shape-memory wire was used for the orthodontic treatment of the anterior teeth in an 11-year-old child who had autism and intellectual disability. The device was designed to reduce the lateral incisor crossbite and the central incisors' labial rotation. The child was treated for 1 year with this removable appliance. Tooth movement was analyzed using cephalograms and surface data were derived from study models. This device proved to be very durable. The lateral incisor crossbite was corrected, and the inclination of the upper central incisors and the interincisal angle were improved. This appliance exerts light and continuous orthodontic force, without requiring any adjustments of the spring wire. The appliance also facilitated orthodontic treatment in a child with intellectual disability in whom treatment with a standard orthodontic device would be unsuitable. ©2012 Special Care Dentistry Association and Wiley Periodicals, Inc.

  16. Effect of pre-strain on microstructure of Ni-Ti orthodontic archwires

    Energy Technology Data Exchange (ETDEWEB)

    Jafari, J. [Department of Materials Science and Engineering, Engineering Faculty, Ferdowsi University of Mashhad, Azadi Square, P.O. Box 91775-1111, Mashhad (Iran, Islamic Republic of); Zebarjad, S.M. [Department of Materials Science and Engineering, Engineering Faculty, Ferdowsi University of Mashhad, Azadi Square, P.O. Box 91775-1111, Mashhad (Iran, Islamic Republic of)], E-mail: Zebarjad@um.ac.ir; Sajjadi, S.A. [Department of Materials Science and Engineering, Engineering Faculty, Ferdowsi University of Mashhad, Azadi Square, P.O. Box 91775-1111, Mashhad (Iran, Islamic Republic of)

    2008-01-25

    One of the most important applications of shape memory alloy is in medicine, especially orthodontic archwires. In this category Ni-Ti orthodontic archwires is one of the oldest used materials. Biocompatibility, corrosion resistance, super elasticity, etc. are its outstanding properties. In spite of the importance of dependency of pre-strain on microstructure of Ni-Ti there are limited sources concentrated on the subject. For this reason the main purpose of the current study is determination of the effect of pre-strain on microstructure of Ni-Ti orthodontic archwires. In this regard, three-point bending was performed on the orthodontic archwire specimens to apply different amount of strain. The microstructures were compared with the un-strained wire using optical and scanning electron microscopes. The results showed that the stable phase depends strongly on the value of pre-strain. Increasing pre-strain causes to decrease martensite laths and leads the microstructure toward austenite phase.

  17. Effect of pre-strain on microstructure of Ni-Ti orthodontic archwires

    International Nuclear Information System (INIS)

    Jafari, J.; Zebarjad, S.M.; Sajjadi, S.A.

    2008-01-01

    One of the most important applications of shape memory alloy is in medicine, especially orthodontic archwires. In this category Ni-Ti orthodontic archwires is one of the oldest used materials. Biocompatibility, corrosion resistance, super elasticity, etc. are its outstanding properties. In spite of the importance of dependency of pre-strain on microstructure of Ni-Ti there are limited sources concentrated on the subject. For this reason the main purpose of the current study is determination of the effect of pre-strain on microstructure of Ni-Ti orthodontic archwires. In this regard, three-point bending was performed on the orthodontic archwire specimens to apply different amount of strain. The microstructures were compared with the un-strained wire using optical and scanning electron microscopes. The results showed that the stable phase depends strongly on the value of pre-strain. Increasing pre-strain causes to decrease martensite laths and leads the microstructure toward austenite phase

  18. Sliding mechanics of coated composite wires and the development of an engineering model for binding.

    Science.gov (United States)

    Zufall, S W; Kusy, R P

    2000-02-01

    A tribological (friction and wear) study, which was designed to simulate clinical sliding mechanics, was conducted as part of an effort to determine the suitability of poly(chloro-p-xylylene) coatings for composite orthodontic archwires. Prototype composite wires, having stiffnesses similar to those of current initial and intermediate alignment wires, were tested against stainless steel and ceramic brackets in the passive and active configurations (with and without angulation). Kinetic coefficient of friction values, which were determined to quantify sliding resistances as functions of the normal forces of ligation, had a mean that was 72% greater than uncoated wire couples at 0.43. To improve analysis of the active configuration, a mathematical model was developed that related bracket angulation, bracket width, interbracket distance, wire geometry, and wire elastic modulus to sliding resistance. From this model, kinetic coefficients of binding were determined to quantify sliding resistances as functions of the normal forces of binding. The mean binding coefficient was the same as that of uncoated wire couples at 0.42. Although penetrations through the coating were observed on many specimens, the glass-fiber reinforcement within the composite wires was undamaged for all conditions tested. This finding implies that the risk of glass fiber release during clinical use would be eliminated by the coating. In addition, the frictional and binding coefficients were still within the limits outlined by conventional orthodontic wire-bracket couples. Consequently, the coatings were regarded as an improvement to the clinical acceptability of composite orthodontic archwires.

  19. Periodontally accelerated osteogenic orthodontics

    Directory of Open Access Journals (Sweden)

    Shailesh Mohanlal Bhandari

    2016-01-01

    Full Text Available This case report demonstrates an orthodontic method that offers short treatment time and the ability to simultaneously reshape and increase the buccolingual thickness of the supporting alveolar bone. A 23-year-old female with Angle′s Class I moderately to severely crowded malocclusion with periodontal pocket with 31 and reduced thickness of the buccal cortical plate of lower anteriors, requested shortened orthodontic treatment time. This surgery technique included buccal full-thickness flaps, selective partial decortication of the cortical plates, concomitant bone grafting/augmentation, and primary flap closure. Following the surgery, orthodontic adjustments were made approximately every 2 weeks. From bracketing to debracketing, the case was completed in approximately 7 months and 3 weeks. Posttreatment evaluation of patient revealed good results. Preexisting thin labial bony cortical plate with lower anteriors was covered. This finding suggests credence to the incorporation of the bone augmentation procedure into the corticotomy surgery because this made it possible to complete the orthodontic treatment with a more intact periodontium. The rapid expansive tooth movements with no significant apical root resorption may be attributed to the osteoclastic or catabolic phase of the regional acceleratory phenomenon.

  20. Cephalometry in orthodontic practice

    International Nuclear Information System (INIS)

    Theunissen, J.J.W.; Terlingen, P.J.A.M.

    1980-01-01

    Cephalometry is applied in orthodontic practice to: - obtain information on the growth of the skull - assist diagnosis of the relationship between the jaws and abnormalities in dentition - evaluate the results of treatment. The course of the diagnostic procedure with the aid of the Ricketts analysis and evaluation of the treatment results using five superimpositions is demonstrated by means of two examples. (Auth.)

  1. Management of gingival recession associated with orthodontic treatment: a case report.

    Science.gov (United States)

    Rana, Tarun Kumar; Phogat, Megha; Sharma, Tarun; Prasad, Narayana; Singh, Shailendra

    2014-07-01

    Many patients undergo orthodontic treatment for aesthetic improvement. It is well established that the patients who undergo orthodontic treatment have a high susceptibility to present plaque accumulation on their teeth because of the presence of brackets, wires and/or other orthodontic elements on the teeth surfaces with which the oral hygiene procedures might be more difficult. The orthodontic treatment is a double-action procedure regarding the periodontal tissues which may be very meaningful in increasing the periodontal health status and may be a harmful procedure which can be followed by several types of periodontal complications. There is a strong correlation between the severity and extent of gingival recessions and the orthodontic treatment suggesting that orthodontic tooth movement may lead to gingival recession. The principal objective in the treatment of gingival recession is to cover the exposed root surfaces to improve aesthetics and to reduce hypersensitivity. Different soft tissue grafting procedures have been proposed in the treatment of gingival recessions. Subepithelial connective tissue graft is a reliable method for treatment of gingival recession. The purpose of this case report was to illustrate the relationship between orthodontic therapy and gingival recession and to describe the management of this case.

  2. Orthodontic-periodontal interactions: Orthodontic extrusion in interdisciplinary regenerative treatments.

    Science.gov (United States)

    Paolone, Maria Giacinta; Kaitsas, Roberto

    2018-06-01

    Orthodontics is a periodontal treatment. "Guided orthodontic regeneration" (GOR) procedures use orthodontic movements in perio-restorative patients. The GOR technique includes a guided orthodontic "soft tissue" regeneration (GOTR) and a guided orthodontic "bone" regeneration (GOBR) with a plastic soft tissue approach and a regenerating reality. The increased amount of soft tissue gained with orthodontic movement can be used for subsequent periodontal regenerative techniques. The increased amount of bone can as well improve primary implant stability and, eventually, simplify a GTR technique to regenerate soft tissues, to restore tooth with external resorption in aesthetic zone or to extract a tooth to create new hard-soft tissue for adjacent teeth. Copyright © 2018. Published by Elsevier Masson SAS.

  3. Aesthetic orthodontic archwires: Progress in their knowledge

    OpenAIRE

    Rongo, Roberto

    2014-01-01

    Introduction: In orthodontics, is quickly increasing the demand for treatments with a very low aesthetic impact in the social life. More and more adult patients want satisfy their necessity to have a beautiful smile, with “invisible” appliances. Numerous are the opportunities to perform an aesthetic orthodontic treatment such as lingual orthodontics, clear aligners or clear labial orthodontics. Aesthetic orthodontic archwires are a component of clear labial orthodontics together with aestheti...

  4. [Expression and purification of a novel thermophilic bacterial single-stranded DNA-binding protein and enhancement the synthesis of DNA and cDNA].

    Science.gov (United States)

    Jia, Xiao-Wei; Zhang, Guo-Hui; Shi, Hai-Yan

    2012-12-01

    Express a novel species of single-stranded DNA-binding protein (SSB) derived from Thermococcus kodakarensis KOD1, abbreviated kod-ssb. And evaluate the effect of kod-ssb on PCR-based DNA amplification and reverse transcription. We express kod-ssb with the Transrtta (DE3), and kod-ssb was purified by affinity chromatography on a Ni2+ Sepharose column, detected by SDS-PAGE. To evaluate the effect of kod-ssb on PCR-based DNA amplification, the human beta globin gene was used as template to amplify a 5-kb, 9-kb and 13-kb. And to detect the effect of kod-ssb on reverse transcription, we used RNA from flu cell culture supernatant extraction as templates to implement qRT-PCR reaction. The plasmid pET11a-kod was transformed into Transetta (DE3) and the recombinant strain Transetta (pET11 a-kod) was obtained. The kod-ssb was highly expressed when the recombinant strain Transetta(pET11a-kod) was induced by IPTG. The specific protein was detected by SDS-PAGE. To confirm that kod-ssb can enhance target DNA synthesis and reduce PCR by-products, 5-, 9-, and 13-kb human beta globin gene fragments were used as templates for PCR. When PCR reactions did not include SSB proteins, the specific PCR product was contaminated with non-specific products. When kod -ssb was added, kod-ssb significantly enhanced amplification of the 5-, 9-and 13-kb target product and minimised the non-specific PCR products. To confirm that kod-ssb can enhance target cDNA synthesis, RNA from flu cell culture supernatant extraction was used as templates for qRT-PCR reaction. The results was that when kod-ssb was added, kod-ssb significantly enhanced the synthesis of cDNA, average Ct value is 19.42, and the average Ct value without kod-ssb is 22.15. kod-ssb may in future be used to enhance DNA and cDNA amplification.

  5. Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans-Expression and characterization.

    Directory of Open Access Journals (Sweden)

    Marcin Olszewski

    Full Text Available DNA polymerases are present in all organisms and are important enzymes that synthesise DNA molecules. They are used in various fields of science, predominantly as essential components for in vitro DNA syntheses, known as PCR. Modern diagnostics, molecular biology and genetic engineering need DNA polymerases which demonstrate improved performance. This study was aimed at obtaining a new NeqSSB-TaqS fusion DNA polymerase from the Taq DNA Stoffel domain and a single-stranded DNA binding-like protein of Nanoarchaeum equitans in order to significantly improve the properties of DNA polymerase. The DNA coding sequence of Taq Stoffel DNA polymerase and the nonspecific DNA-binding protein of Nanoarchaeum equitans (NeqSSB-like protein were fused. A novel recombinant gene was obtained which was cloned into the pET-30 Ek/LIC vector and introduced into E. coli for expression. The recombinant enzyme was purified and its enzymatic properties including DNA polymerase activity, PCR amplification rate, thermostability, processivity and resistance to inhibitors, were tested. The yield of the target protein reached approximately 18 mg/l after 24 h of the IPTG induction. The specific activity of the polymerase was 2200 U/mg. The recombinant NeqSSB-TaqS exhibited a much higher extension rate (1000 bp template in 20 s, processivity (19 nt, thermostability (half-life 35 min at 95°C and higher tolerance to PCR inhibitors (0.3-1.25% of whole blood, 0.84-13.5 μg of lactoferrin and 4.7-150 ng of heparin than Taq Stoffel DNA polymerase. Furthermore, our studies show that NeqSSB-TaqS DNA polymerase has a high level of flexibility in relation to Mg2+ ions (from 1 to 5 mM and KCl or (NH42SO4 salts (more than 60 mM and 40 mM, respectively. Using NeqSSB-TaqS DNA polymerase instead of the Taq DNA polymerase could be a better choice in many PCR applications.

  6. Single strand conformation polymorphism based SNP and Indel markers for genetic mapping and synteny analysis of common bean (Phaseolus vulgaris L.

    Directory of Open Access Journals (Sweden)

    Gómez Marcela

    2009-12-01

    Full Text Available Abstract Background Expressed sequence tags (ESTs are an important source of gene-based markers such as those based on insertion-deletions (Indels or single-nucleotide polymorphisms (SNPs. Several gel based methods have been reported for the detection of sequence variants, however they have not been widely exploited in common bean, an important legume crop of the developing world. The objectives of this project were to develop and map EST based markers using analysis of single strand conformation polymorphisms (SSCPs, to create a transcript map for common bean and to compare synteny of the common bean map with sequenced chromosomes of other legumes. Results A set of 418 EST based amplicons were evaluated for parental polymorphisms using the SSCP technique and 26% of these presented a clear conformational or size polymorphism between Andean and Mesoamerican genotypes. The amplicon based markers were then used for genetic mapping with segregation analysis performed in the DOR364 × G19833 recombinant inbred line (RIL population. A total of 118 new marker loci were placed into an integrated molecular map for common bean consisting of 288 markers. Of these, 218 were used for synteny analysis and 186 presented homology with segments of the soybean genome with an e-value lower than 7 × 10-12. The synteny analysis with soybean showed a mosaic pattern of syntenic blocks with most segments of any one common bean linkage group associated with two soybean chromosomes. The analysis with Medicago truncatula and Lotus japonicus presented fewer syntenic regions consistent with the more distant phylogenetic relationship between the galegoid and phaseoloid legumes. Conclusion The SSCP technique is a useful and inexpensive alternative to other SNP or Indel detection techniques for saturating the common bean genetic map with functional markers that may be useful in marker assisted selection. In addition, the genetic markers based on ESTs allowed the construction

  7. Single strand conformation polymorphism based SNP and Indel markers for genetic mapping and synteny analysis of common bean (Phaseolus vulgaris L.).

    Science.gov (United States)

    Galeano, Carlos H; Fernández, Andrea C; Gómez, Marcela; Blair, Matthew W

    2009-12-23

    Expressed sequence tags (ESTs) are an important source of gene-based markers such as those based on insertion-deletions (Indels) or single-nucleotide polymorphisms (SNPs). Several gel based methods have been reported for the detection of sequence variants, however they have not been widely exploited in common bean, an important legume crop of the developing world. The objectives of this project were to develop and map EST based markers using analysis of single strand conformation polymorphisms (SSCPs), to create a transcript map for common bean and to compare synteny of the common bean map with sequenced chromosomes of other legumes. A set of 418 EST based amplicons were evaluated for parental polymorphisms using the SSCP technique and 26% of these presented a clear conformational or size polymorphism between Andean and Mesoamerican genotypes. The amplicon based markers were then used for genetic mapping with segregation analysis performed in the DOR364 x G19833 recombinant inbred line (RIL) population. A total of 118 new marker loci were placed into an integrated molecular map for common bean consisting of 288 markers. Of these, 218 were used for synteny analysis and 186 presented homology with segments of the soybean genome with an e-value lower than 7 x 10-12. The synteny analysis with soybean showed a mosaic pattern of syntenic blocks with most segments of any one common bean linkage group associated with two soybean chromosomes. The analysis with Medicago truncatula and Lotus japonicus presented fewer syntenic regions consistent with the more distant phylogenetic relationship between the galegoid and phaseoloid legumes. The SSCP technique is a useful and inexpensive alternative to other SNP or Indel detection techniques for saturating the common bean genetic map with functional markers that may be useful in marker assisted selection. In addition, the genetic markers based on ESTs allowed the construction of a transcript map and given their high conservation

  8. Defibrotide, a polydisperse mixture of single-stranded phosphodiester oligonucleotides with lifesaving activity in severe hepatic veno-occlusive disease: clinical outcomes and potential mechanisms of action.

    Science.gov (United States)

    Kornblum, Noah; Ayyanar, Kanyalakshmi; Benimetskaya, Luba; Richardson, Paul; Iacobelli, Massimo; Stein, C A

    2006-01-01

    Veno-occlusive disease of the liver (VOD) remains a troubling and potentially fatal complication of high-dose chemotherapy and hematopoietic stem cell transplantation conditioning regimens. No effective therapy has been available for these patients to date, and the best supportive care measures remain woefully inadequate. Defibrotide (DF) (Gentium, S.p.A., Como, Italy), a polydisperse mixture of all the single-stranded phosphodiester oligodeoxyribonucleotides that can be obtained from the controlled depolymerization of porcine intestinal mucosal genomic DNA, seems to offer a safe and effective treatment for some patients suffering from severe VOD, a condition for which no accepted standard therapy currently exists. Early clinical studies evaluating the efficacy of DF for the treatment of severe VOD in patients undergoing hematopoietic stem cell transplantation have been very encouraging. Approximately 45% of the patients treated in multiple initial phase II clinical trials achieved a complete response at day +100, demonstrating normalization of serum bilirubin and resolution of the clinical syndrome. However, although multi-institutional, these represented single arm studies. A large, FDA-approved, pivotal, prospective, multi-institutional, global phase III trial of DF vs. historical controls (best available therapy) commenced in the first quarter of 2006 and should provide further validation of DF's efficacy. The drug seems to have few significant side effects, and almost all test subjects who have received this treatment have tolerated it well. Although the mechanism of action remains unclear, the drug exerts minimal systemic anticoagulant effects yet appears to induce numerous antithrombotic and profibrinolytic effects both in vitro and in vivo. It may function as an adenosine receptor agonist and causes increased concentrations of endogenous prostaglandins, which modulate thrombomodulin, platelets, and fibrinolysis. It also appears to block lipopolysaccharide

  9. Contribution of single-strand breaks and alkali-labile bonds to the loss of infectivity of γ-irradiated phiX174 RF-DNA in E. coli cells mutant in various repair functions

    International Nuclear Information System (INIS)

    McKee, R.H.

    1975-01-01

    Twenty-one radiation sensitive mutants have been examined for their capacity to support gamma-irradiated phiX174 RF-DNA. The survival of phiX174 RF-DNA was reduced in essentially all of the sensitive mutants. The irradiated phiX174 RF-DNA was then separated into populations containing either single-strand breaks or alkali-labile bonds to examine the capacity of the mutants to repair each of the classes of lesions. It was found that all E. coli strains are unable to repair 22 percent of the single-strand breaks and all sensitive mutants are unable to repair an additional 10 percent of the breaks. All the repair functions examined are involved in single-strand break repair and none are more or less necessary than any of the others. PhiX174 RF-DNA is also inactivated by alkali-labile bonds. In the normal strains the inactivation efficiency is 0.16 lethal events per lesion with a threshold dose of 15 to 20 krads. The mutants are divided into two classes by their sensitivity to alkali-labile bonds. Both classes of mutants are also inactivated by alkali-labile bonds with efficiencies of about 0.17 and 0.29 lethal events per lesion, respectively. It is proposed that the differences seen in survival curves of phiX174 measured in the sensitive mutants is due to this difference. Although in normal cells the efficiency of inactivation of phiX174 by single-strand breaks is 50 percent greater than by alkali-labile bonds, alkali-labile bonds are produced at approximately twice the rate of single-strand breaks so alkali-labile bonds account for about 61 percent of the overall inactivation. In the mutants of least sensitivity alkali-labile bonds account for about 54 percent of the inactivating events and in the most sensitive about 67 percent

  10. Experimental testing and modelling of a resistive type superconducting fault current limiter using MgB2 wire

    International Nuclear Information System (INIS)

    Smith, A C; Pei, X; Oliver, A; Husband, M; Rindfleisch, M

    2012-01-01

    A prototype resistive superconducting fault current limiter (SFCL) was developed using single-strand round magnesium diboride (MgB 2 ) wire. The MgB 2 wire was wound with an interleaved arrangement to minimize coil inductance and provide adequate inter-turn voltage withstand capability. The temperature profile from 30 to 40 K and frequency profile from 10 to 100 Hz at 25 K were tested and reported. The quench properties of the prototype coil were tested using a high current test circuit. The fault current was limited by the prototype coil within the first quarter-cycle. The prototype coil demonstrated reliable and repeatable current limiting properties and was able to withstand a potential peak current of 372 A for one second without any degradation of performance. A three-strand SFCL coil was investigated and demonstrated scaled-up current capacity. An analytical model to predict the behaviour of the prototype single-strand SFCL coil was developed using an adiabatic boundary condition on the outer surface of the wire. The predicted fault current using the analytical model showed very good correlation with the experimental test results. The analytical model and a finite element thermal model were used to predict the temperature rise of the wire during a fault. (paper)

  11. Biodegradation of orthodontic metallic brackets and associated implications for friction.

    Science.gov (United States)

    Regis, Saulo; Soares, Paulo; Camargo, Elisa S; Guariza Filho, Odilon; Tanaka, Orlando; Maruo, Hiroshi

    2011-10-01

    This study aimed to assess the effect of clinical exposure on the surface morphology, dimensions, and frictional behavior of metallic orthodontic brackets. Ninety-five brackets, of 3 commercial brands, were retrieved from patients who had finished orthodontic treatment. As-received brackets, matched by type and brand, were used for comparisons. Surface morphology and precipitated material were analyzed by optical and scanning electron microscopy and x-ray microanalysis. Bracket dimensions were measured with a measuring microscope. Resistance to sliding on a stainless steel wire was assessed. Retrieved brackets showed surface alterations from corrosion, wear, and plastic deformation, especially in the external slot edges. Film deposition over the alloy surface was observed to a variable extent. The main elements in the film were carbon, oxygen, calcium, and phosphorus. The as-received brackets showed differences (P brackets' slots. The frictional behavior differed among brands. Retrieved brackets of 2 brands showed 10% to 20% increases in resistance to sliding. Metallic brackets undergo significant degradation during orthodontic treatment, possibly with increased friction. At present, it is difficult to predict the impact of these changes on the clinical performance of orthodontic components. Copyright © 2011 American Association of Orthodontists. Published by Mosby, Inc. All rights reserved.

  12. Corrosion of weldments in orthodontic appliances

    International Nuclear Information System (INIS)

    Vazquez, S.M.; Riesgo, O.; Duffo, G.S.

    1997-01-01

    The study of corrosion-related problems of dental materials has undergone a considerable development in recent years in order to avoid the use of materials with insufficient corrosion resistance in patients mouth. The subject of the present work was to study a particular type of corrosion: galvanic corrosion. One of the most common case of galvanic couples in patients mouth are the orthodontic appliances. The materials studied in the present work were stainless steel strips and silver-copper wires, isolated and welded between them. The electrochemical tests were performed in a NaCl 0.1M and Lactic Acid 0.1 M solution (pH2.3), and after test, the specimens were observed using the optical and scanning electron microscope. The results show that when stainless steel is coupled with a silver solder, the last is the anode of the galvanic couple. As a consequence of this, the silver solder undergone a severe attack. Stainless steel orthodontic appliances with silver solder are feasibly destroyed due to a severe attack on the filler metal disjoining the welded parts. (author) 9 refs

  13. Optimal management of orthodontic pain.

    Science.gov (United States)

    Topolski, Francielle; Moro, Alexandre; Correr, Gisele Maria; Schimim, Sasha Cristina

    2018-01-01

    Pain is an undesirable side effect of orthodontic tooth movement, which causes many patients to give up orthodontic treatment or avoid it altogether. The aim of this study was to investigate, through an analysis of the scientific literature, the best method for managing orthodontic pain. The methodological aspects involved careful definition of keywords and diligent search in databases of scientific articles published in the English language, without any restriction of publication date. We recovered 1281 articles. After the filtering and classification of these articles, 56 randomized clinical trials were selected. Of these, 19 evaluated the effects of different types of drugs for the control of orthodontic pain, 16 evaluated the effects of low-level laser therapy on orthodontic pain, and 21 evaluated other methods of pain control. Drugs reported as effective in orthodontic pain control included ibuprofen, paracetamol, naproxen sodium, aspirin, etoricoxib, meloxicam, piroxicam, and tenoxicam. Most studies report favorable outcomes in terms of alleviation of orthodontic pain with the use of low-level laser therapy. Nevertheless, we noticed that there is no consensus, both for the drug and for laser therapy, on the doses and clinical protocols most appropriate for orthodontic pain management. Alternative methods for orthodontic pain control can also broaden the clinician's range of options in the search for better patient care.

  14. Craniofacial orthodontics and postgraduate orthodontic training in Nigeria.

    Science.gov (United States)

    Isiekwe, G I; Oguchi, C O; daCosta, O O; Utomi, I L

    2016-01-01

    Craniofacial orthodontics has been shown to be a critical component of the care of patients with craniofacial anomalies such as cleft lip and palate. Thus, the purpose of this study was to assess the perceptions and clinical experience in cleft and craniofacial care, of orthodontic residents in Nigeria. Questionnaires were sent out to orthodontic residents in the six Postgraduate Orthodontic Training Centers in the country at that time. The questionnaires were self-administered and covered areas in beliefs in cleft care and the clinical experience and challenges faced by the residents in the provision of craniofacial orthodontic care at their various institutions. Thirty-three respondents returned completed questionnaires, with a response rate of 97%. All the respondents believed that residents should be involved in cleft and craniofacial care. Postnatal counseling was the clinical procedure in which the residents reported the highest level of clinical experience (47.4%). The least clinical experience was recorded in pre-bone graft orthodontics (7.4%) and orthodontic preparation for orthognathic surgery (5.5%). Some of the challenges highlighted by the residents were low patients turn out for orthodontic care and the absence of multidisciplinary treatment for craniofacial patients in their centers. Orthodontic residents in Nigeria believe that they should be involved in the management of patients with craniofacial anomalies and cleft lip and palate. However, majority of the residents have limited clinical experience in the management of these patients. A lot more needs to be done, to expose orthodontic residents in training, to all aspects of the orthodontic and multidisciplinary team care required for the cleft/craniofacial patient.

  15. Improvement of orthodontic friction by coating archwire with carbon nitride film

    International Nuclear Information System (INIS)

    Wei Songbo; Shao Tianmin; Ding Peng

    2011-01-01

    In order to reduce frictional resistance between archwire and bracket during orthodontic tooth movement, carbon nitride (CNx) thin films were deposited on the surface of archwires with ion beam assisted deposition (IBAD). The energy-dispersive X-ray spectrometer (EDS) analysis showed that the CNx film was successfully deposited on the surface of the orthodontic wires. X-ray photoelectron spectroscopy (XPS) analysis suggested that the deposited CNx film was sp 2 carbon dominated structures, and diversiform bonds (N-C, N≡C, et al.) coexisted in the film. The friction tests indicated that the CNx film significantly reduced the wire-bracket friction both in ambient air and in artificial saliva. The sp 2 C rich structure of the CNx film as well as its protection function for the archwire was responsible for the low friction of the wire-bracket sliding system.

  16. Improvement of orthodontic friction by coating archwire with carbon nitride film

    Energy Technology Data Exchange (ETDEWEB)

    Wei Songbo [State Key Laboratory of Tribology, Tsinghua University, Beijing 100084 (China); Shao Tianmin, E-mail: shaotm@mail.tsinghua.edu.cn [State Key Laboratory of Tribology, Tsinghua University, Beijing 100084 (China); Ding Peng [Department of Orthodontics, Peking University School and Hospital of Stomatology, Beijing 100081 (China)

    2011-10-01

    In order to reduce frictional resistance between archwire and bracket during orthodontic tooth movement, carbon nitride (CNx) thin films were deposited on the surface of archwires with ion beam assisted deposition (IBAD). The energy-dispersive X-ray spectrometer (EDS) analysis showed that the CNx film was successfully deposited on the surface of the orthodontic wires. X-ray photoelectron spectroscopy (XPS) analysis suggested that the deposited CNx film was sp{sup 2} carbon dominated structures, and diversiform bonds (N-C, N{identical_to}C, et al.) coexisted in the film. The friction tests indicated that the CNx film significantly reduced the wire-bracket friction both in ambient air and in artificial saliva. The sp{sup 2}C rich structure of the CNx film as well as its protection function for the archwire was responsible for the low friction of the wire-bracket sliding system.

  17. The influence of different types of fixed orthodontic appliance on the growth and adherence of microorganisms (in vitro study).

    Science.gov (United States)

    Saloom, Hayder F; Mohammed-Salih, Harraa S; Rasheed, Shaymaa F

    2013-02-01

    Orthodontic appliances serve as different impact zones and modify microbial adherence and colonization, acting as foreign reserves and possible sources of infection. This study was conducted to investigate the effect of different types of fixed orthodontic appliances on the growth and adherence of microorganisms in oral flora which are Streptococcus mutans (S. mutans) and Candida albicans. Sixty-four of four different fixed orthodontic appliance-samples were used, divided into four groups of sixteen. Type I: Sapphire brackets- Coated wires, type II: Sapphire brackets- Stainless steel wires, type III: Stainless steel brackets- Coated wires and type IV: Stainless steel brackets- Stainless steel wires. Oral strains of S. mutans and Candida albicans were studied in the present study using biochemical test then microbial suspensions were prepared to do the tests of each microorganism including the antimicrobial effects of different appliance-samples on the growth of microorganisms and their adhesion tests. The results showed significant differences between the different appliances in terms of inhibition zone formation (PAppliance with high esthetic appearance, sapphire brackets and coated arch wire, showed the least adherence of S. mutans and Candida albicans in comparison to other appliances with less esthetic and more metal components. Key words:Orthodontic appliance, Adherence, Streptococcus mutans, Candida albicans.

  18. Laser Applications in Orthodontics

    Science.gov (United States)

    Heidari, Somayeh; Torkan, Sepideh

    2013-01-01

    A laser is a collimated single wavelength of light which delivers a concentrated source of energy. Soon after different types of lasers were invented, investigators began to examine the effects of different wavelengths of laser energy on oral tissues, routine dental procedures and experimental applications. Orthodontists, along with other specialist in different fields of dentistry, can now benefit from several different advantages that lasers provide during the treatment process, from the beginning of the treatment, when separators are placed, to the time of resin residues removal from the tooth surface at the end of orthodontic treatment. This article outlines some of the most common usages of laser beam in orthodontics and also provides a comparison between laser and other conventional method that were the standard of care prior to the advent of laser in this field. PMID:25606324

  19. Optimal management of orthodontic pain

    Directory of Open Access Journals (Sweden)

    Topolski F

    2018-03-01

    Full Text Available Francielle Topolski,1 Alexandre Moro,1,2 Gisele Maria Correr,3 Sasha Cristina Schimim1 1Department of Orthodontics, Positivo University, Curitiba, Paraná, Brazil; 2Department of Orthodontics, Federal University of Paraná, Curitiba, Paraná, Brazil; 3Department of Restorative Dentistry, Positivo University, Curitiba, Paraná, Brazil Abstract: Pain is an undesirable side effect of orthodontic tooth movement, which causes many patients to give up orthodontic treatment or avoid it altogether. The aim of this study was to investigate, through an analysis of the scientific literature, the best method for managing orthodontic pain. The methodological aspects involved careful definition of keywords and diligent search in databases of scientific articles published in the English language, without any restriction of publication date. We recovered 1281 articles. After the filtering and classification of these articles, 56 randomized clinical trials were selected. Of these, 19 evaluated the effects of different types of drugs for the control of orthodontic pain, 16 evaluated the effects of low-level laser therapy on orthodontic pain, and 21 evaluated other methods of pain control. Drugs reported as effective in orthodontic pain control included ibuprofen, paracetamol, naproxen sodium, aspirin, etoricoxib, meloxicam, piroxicam, and tenoxicam. Most studies report favorable outcomes in terms of alleviation of orthodontic pain with the use of low-level laser therapy. Nevertheless, we noticed that there is no consensus, both for the drug and for laser therapy, on the doses and clinical protocols most appropriate for orthodontic pain management. Alternative methods for orthodontic pain control can also broaden the clinician’s range of options in the search for better patient care. Keywords: tooth movement, pain control, drug therapy, laser therapy

  20. Oral health: orthodontic treatment.

    Science.gov (United States)

    Martonffy, Andrea Ildiko

    2015-01-01

    Improper tooth alignment due to crowding, malocclusion, and missing teeth can cause difficulties with eating and speech, and premature wear. It is estimated that more than 20% of children would benefit from orthodontic treatment to correct these conditions, many of which will persist into adulthood if not corrected. Orthodontic care is gaining popularity among adults for similar concerns, as well as for correction of cosmetic issues. The psychological effects of malocclusion should not be ignored. The American Association of Orthodontists recommends that all children undergo evaluation at the first recognition of an orthodontic condition and no later than age 7 years. Some children will need early treatment to help eliminate developing conditions and improve the foundations of the bite, which can ease later treatment in adolescence. For others, treatment in adolescence without early treatment is recommended. Standard cemented braces or clear, removable aligners may be used, depending on the patient's corrective needs. Average treatment time is approximately 2 years; this may be shortened by the use of accelerative techniques. Routine preventive dental care should be continued during the treatment period. Written permission from the American Academy of Family Physicians is required for reproduction of this material in whole or in part in any form or medium.

  1. A comparative study of metal artifacts from common metal orthodontic brackets in magnetic resonance imaging

    International Nuclear Information System (INIS)

    Kajan, Zahra Dalili; Alizadeh, Ahmad; Hemmaty, Yasmin Babael; Roushan, Zahra Atrkar; Khademi, Jalil

    2015-01-01

    This study was performed to compare the metal artifacts from common metal orthodontic brackets in magnetic resonance imaging. A dry mandible with 12 intact premolars was prepared, and was scanned ten times with various types of brackets: American, 3M, Dentaurum, and Masel orthodontic brackets were used, together with either stainless steel (SS) or nickel titanium (NiTi) wires. Subsequently, three different sequences of coronal and axial images were obtained: spin-echo T1-weighted images, fast spin-echo T2-weighted images, and fluid-attenuated inversion recovery images. In each sequence, the two sequential axial and coronal images with the largest signal-void area were selected. The largest diameters of the signal voids in the direction of the X-, Y-, and Z-axes were then measured twice. Finally, the mean linear values associated with different orthodontic brackets were analyzed using one-way analysis of variation, and the results were compared using the independent t-test to assess whether the use of SS or NiTi wires had a significant effect on the images. Statistically significant differences were only observed along the Z-axis among the four different brands of orthodontic brackets with SS wires. A statistically significant difference was observed along all axes among the brackets with NiTi wires. A statistically significant difference was found only along the Z-axis between nickel-free and nickel-containing brackets. With respect to all axes, the 3M bracket was associated with smaller signal-void areas. Overall, the 3M and Dentaurum brackets with NiTi wires induced smaller artifacts along all axes than those with SS wires

  2. A comparative study of metal artifacts from common metal orthodontic brackets in magnetic resonance imaging

    Energy Technology Data Exchange (ETDEWEB)

    Kajan, Zahra Dalili; Alizadeh, Ahmad; Hemmaty, Yasmin Babael; Roushan, Zahra Atrkar; Khademi, Jalil [Guilan University of Medical Sciences, Rasht (Iran, Islamic Republic of)

    2015-09-15

    This study was performed to compare the metal artifacts from common metal orthodontic brackets in magnetic resonance imaging. A dry mandible with 12 intact premolars was prepared, and was scanned ten times with various types of brackets: American, 3M, Dentaurum, and Masel orthodontic brackets were used, together with either stainless steel (SS) or nickel titanium (NiTi) wires. Subsequently, three different sequences of coronal and axial images were obtained: spin-echo T1-weighted images, fast spin-echo T2-weighted images, and fluid-attenuated inversion recovery images. In each sequence, the two sequential axial and coronal images with the largest signal-void area were selected. The largest diameters of the signal voids in the direction of the X-, Y-, and Z-axes were then measured twice. Finally, the mean linear values associated with different orthodontic brackets were analyzed using one-way analysis of variation, and the results were compared using the independent t-test to assess whether the use of SS or NiTi wires had a significant effect on the images. Statistically significant differences were only observed along the Z-axis among the four different brands of orthodontic brackets with SS wires. A statistically significant difference was observed along all axes among the brackets with NiTi wires. A statistically significant difference was found only along the Z-axis between nickel-free and nickel-containing brackets. With respect to all axes, the 3M bracket was associated with smaller signal-void areas. Overall, the 3M and Dentaurum brackets with NiTi wires induced smaller artifacts along all axes than those with SS wires.

  3. A comparative study of metal artifacts from common metal orthodontic brackets in magnetic resonance imaging.

    Science.gov (United States)

    Dalili Kajan, Zahra; Khademi, Jalil; Alizadeh, Ahmad; Babaei Hemmaty, Yasamin; Atrkar Roushan, Zahra

    2015-09-01

    This study was performed to compare the metal artifacts from common metal orthodontic brackets in magnetic resonance imaging. A dry mandible with 12 intact premolars was prepared, and was scanned ten times with various types of brackets: American, 3M, Dentaurum, and Masel orthodontic brackets were used, together with either stainless steel (SS) or nickel titanium (NiTi) wires. Subsequently, three different sequences of coronal and axial images were obtained: spin-echo T1 -weighted images, fast spin-echo T2 -weighted images, and fluid-attenuated inversion recovery images. In each sequence, the two sequential axial and coronal images with the largest signal-void area were selected. The largest diameters of the signal voids in the direction of the X-, Y-, and Z-axes were then measured twice. Finally, the mean linear values associated with different orthodontic brackets were analyzed using one-way analysis of variation, and the results were compared using the independent t-test to assess whether the use of SS or NiTi wires had a significant effect on the images. Statistically significant differences were only observed along the Z-axis among the four different brands of orthodontic brackets with SS wires. A statistically significant difference was observed along all axes among the brackets with NiTi wires. A statistically significant difference was found only along the Z-axis between nickel-free and nickel-containing brackets. With respect to all axes, the 3M bracket was associated with smaller signal-void areas. Overall, the 3M and Dentaurum brackets with NiTi wires induced smaller artifacts along all axes than those with SS wires.

  4. Effects of different brazing and welding methods on the fracture load of various orthodontic joining configurations.

    Science.gov (United States)

    Bock, Jens J; Bailly, Jacqueline; Fuhrmann, Robert A

    2009-06-01

    The aim of this study was to compare the fracture load of different joints made by conventional brazing, tungston inert gas (TIG) and laser welding. Six standardized joining configurations of spring hard quality orthodontic wire were investigated: end-to-end, round, cross, 3 mm length, 9 mm length and 6.5 mm to orthodontic band. The joints were made by five different methods: brazing with universal silver solder, two TIG and two laser welding devices. The fracture loads were measured with a universal testing machine (Zwick 005). Data were analysed with the Mann-Whitney-Wilcoxon and Kruskal-Wallis tests. The significance level was set at Pwelding (Pwelding (826 N). No differences between the various TIG or laser welding devices were demonstrated, although it was not possible to join an orthodontic wire to an orthodontic band using TIG welding. For orthodontic purposes laser and TIG welding are solder free alternatives. TIG welding and laser welding showed similar results. The laser technique is an expensive, but sophisticated and simple method.

  5. Application of irradiated wire

    International Nuclear Information System (INIS)

    Uda, I.; Kozima, K.; Suzuki, S.; Tada, S.; Torisu, S.; Veno, K.

    1984-01-01

    Rubber insulated wires are still useful for internal wiring in motor vehicles and electrical equipment because of flexibility and toughness. Irradiated cross-linked rubber materials have been successfully introduced for use with fusible link wire and helically coiled cord

  6. Noncontextual Wirings

    Science.gov (United States)

    Amaral, Barbara; Cabello, Adán; Cunha, Marcelo Terra; Aolita, Leandro

    2018-03-01

    Contextuality is a fundamental feature of quantum theory necessary for certain models of quantum computation and communication. Serious steps have therefore been taken towards a formal framework for contextuality as an operational resource. However, the main ingredient of a resource theory—a concrete, explicit form of free operations of contextuality—was still missing. Here we provide such a component by introducing noncontextual wirings: a class of contextuality-free operations with a clear operational interpretation and a friendly parametrization. We characterize them completely for general black-box measurement devices with arbitrarily many inputs and outputs. As applications, we show that the relative entropy of contextuality is a contextuality monotone and that maximally contextual boxes that serve as contextuality bits exist for a broad class of scenarios. Our results complete a unified resource-theoretic framework for contextuality and Bell nonlocality.

  7. Reduction in static friction by deposition of a homogeneous diamond-like carbon (DLC) coating on orthodontic brackets.

    Science.gov (United States)

    Akaike, Shun; Hayakawa, Tohru; Kobayashi, Daishiro; Aono, Yuko; Hirata, Atsushi; Hiratsuka, Masanori; Nakamura, Yoshiki

    2015-01-01

    In orthodontics, a reduction in static friction between the brackets and wire is important to enable easy tooth movement. The aim of this study was to examine the effects of a homogeneous diamond-like carbon (DLC) coating on the whole surfaces of slots in stainless steel orthodontic brackets on reducing the static friction between the brackets and the wire. The DLC coating was characterized using Raman spectroscopy, surface roughness and contact angle measurements, and SEM observations. Rectangular stainless steel and titanium-molybdenum alloy wires with two different sizes were employed, and the static friction between the brackets and wire was measured under dry and wet conditions. The DLC coating had a thickness of approximately 1.0 μm and an amorphous structure was identified. The results indicated that the DLC coating always led to a reduction in static friction.

  8. Periodontic and orthodontic treatment in adults.

    Science.gov (United States)

    Ong, Marianne M A; Wang, Hom-Lay

    2002-10-01

    The purpose of this article is to provide an update of the interrelationship between periodontics and orthodontics in adults. Specific areas reviewed are the reaction of periodontal tissue to orthodontic forces, the influence of tooth movement on the periodontium, the effect of circumferential supracrestal fiberotomy in preventing orthodontic relapse, the effect of orthodontic treatment on the periodontium, microbiology associated with orthodontic bands, and mucogingival and esthetic considerations. In addition, the relationship between orthodontics and implants (eg, using dental implants for orthodontic anchorage) is discussed.

  9. The differences of orthodontic tooth movement on menstrual and ovulation cycle

    Directory of Open Access Journals (Sweden)

    Sonya Grecila Susilo

    2014-12-01

    Full Text Available Background: Estrogens are sex hormon that play an important role in bone metabolism, including in bone remodeling during orthodontic treatment. Women has a monthly cycle which is affected by fluctuations of estrogen that is menstruation and ovulation. Purpose: The study was aimed to determine the differences of orthodontic tooth movement during menstrual an ovulation cycle. Methods: Five women were given ± 100 g orthodontic force using fixed orthodontic appliance with straight wire technique at the time of menstruation and ovulation with an interval of 1.5 months. Orthodontic tooth movement and levels of estrogen were measured during menstruation and ovulation. Results: Statistical results showed a significant differences between estrogen levels and orthodontic tooth movement (p < 0.05. When estrogen levels decline as in menstruation, tooth movement as results of orthodontic force would increase, whereas when estrogen levels increase as the time of ovulation, tooth movement would decrease. Conclusion: The estrogen level in menstruation and ovulation cycle may affect the tooth movement on ortodontic treatment.Latar belakang: Estrogen merupakan hormon seks yang mempunyai peran penting dalam metabolisme tulang termasuk dalam remodeling tulang selama perawatan ortodonti. Wanita memiliki siklus bulanan yang dipengaruhi oleh fluktuasi estrogen yaitu menstruasi dan ovulasi. Tujuan: Penelitian ini bertujuan untuk mengetahui perbedaan pergerakan gigi akibat gaya ortodonti pada siklus menstruasi dan ovulasi. Metode: Lima orang wanita diberikan gaya ortodonti ±100 gr menggunakan alat orthodontik cekat dengan teknik straight wire pada saat menstruasi dan ovulasi dengan selang waktu 1,5 bulan. Selain itu subyek juga diukur kadar estrogennya saat menstruasi dan ovulasi. Hasil: Hasil statistik menunjukan kadar estrogen berbanding terbalik dengan pergerakan gigi ortodonti (p < 0,05. Saat kadar estrogen menurun seperti pada menstruasi, maka pergerakan gigi

  10. Lingual orthodontic education: An insight

    Directory of Open Access Journals (Sweden)

    Surya Kanta Das

    2016-01-01

    Full Text Available Despite increasing demand for lingual orthodontics, the technique is not very popular among the orthodontists in general. Lingual orthodontics differs from the conventional labial technique in all aspects. Lack of comprehensive training in this field is a major obstacle in popularizing this science of invisible orthodontics. At present, short-term courses and part-time degree programs are the means to learn this technique and the demand for more comprehensive lingual orthodontic education is on a rise among orthodontists. Lingual orthodontics as a super specialty discipline with full-time residency program can be a step forward. This will groom orthodontists to acquire the finest skills to finish lingual cases but also help to the science to grow with dedicated research work.

  11. Clear aligners in orthodontic treatment.

    Science.gov (United States)

    Weir, T

    2017-03-01

    Since the introduction of the Tooth Positioner (TP Orthodontics) in 1944, removable appliances analogous to clear aligners have been employed for mild to moderate orthodontic tooth movements. Clear aligner therapy has been a part of orthodontic practice for decades, but has, particularly since the introduction of Invisalign appliances (Align Technology) in 1998, become an increasingly common addition to the orthodontic armamentarium. An internet search reveals at least 27 different clear aligner products currently on offer for orthodontic treatment. The present paper will highlight the increasing popularity of clear aligner appliances, as well as the clinical scope and the limitations of aligner therapy in general. Further, the paper will outline the differences between the various types of clear aligner products currently available. © 2017 Australian Dental Association.

  12. The application of strand invasion phenomenon, directed by peptide nucleic acid (PNA) and single-stranded DNA binding protein (SSB) for the recognition of specific sequences of human endogenous retroviral HERV-W family.

    Science.gov (United States)

    Machnik, Grzegorz; Bułdak, Łukasz; Ruczyński, Jarosław; Gąsior, Tomasz; Huzarska, Małgorzata; Belowski, Dariusz; Alenowicz, Magdalena; Mucha, Piotr; Rekowski, Piotr; Okopień, Bogusław

    2017-05-01

    The HERV-W family of human endogenous retroviruses represents a group of numerous sequences that show close similarity in genetic composition. It has been documented that some members of HERV-W-derived expression products are supposed to play significant role in humans' pathology, such as multiple sclerosis or schizophrenia. Other members of the family are necessary to orchestrate physiological processes (eg, ERVWE1 coding syncytin-1 that is engaged in syncytiotrophoblast formation). Therefore, an assay that would allow the recognition of particular form of HERV-W members is highly desirable. A peptide nucleic acid (PNA)-mediated technique for the discrimination between multiple sclerosis-associated retrovirus and ERVWE1 sequence has been developed. The assay uses a PNA probe that, being fully complementary to the ERVWE1 but not to multiple sclerosis-associated retrovirus (MSRV) template, shows high selective potential. Single-stranded DNA binding protein facilitates the PNA-mediated, sequence-specific formation of strand invasion complex and, consequently, local DNA unwinding. The target DNA may be then excluded from further analysis in any downstream process such as single-stranded DNA-specific exonuclease action. Finally, the reaction conditions have been optimized, and several PNA probes that are targeted toward distinct loci along whole HERV-W env sequences have been evaluated. We believe that PNA/single-stranded DNA binding protein-based application has the potential to selectively discriminate particular HERV-W molecules as they are at least suspected to play pathogenic role in a broad range of medical conditions, from psycho-neurologic disorders (multiple sclerosis and schizophrenia) and cancers (breast cancer) to that of an auto-immunologic background (psoriasis and lupus erythematosus). Copyright © 2016 John Wiley & Sons, Ltd.

  13. Application of Single Strand Conformational Polymorphism (PCR-SSCP) in Identification of Some Beta-Globin Gene Mutations in A Group of Egyptian Beta-Thalassemia Patients and Carriers

    International Nuclear Information System (INIS)

    Somaya, E.T.; Soliman, M.D

    2010-01-01

    The present study investigated whether the single-strand conformational polymorphism (SSCP) method could be employed to identify (rather than simply detect) four of the most common beta-globin gene mutations in the Egyptian population: IVS-I-110, IVS-I-6, the IVS-I-1, and Codon 39. Using DNA from 90 beta-thalassemia patients and carriers, by PCR the appropriate 238-bp region of the human beta-globin gene was amplified, the reaction products (Single-stranded DNA) were analyzed by none denaturing polyacrylamide gel electrophoresis, and the bands visualized by silver staining. Single-stranded DNA (ssDNA) fragments showed reproducible pattern of bands that were characteristic of the mutations present. With the use of control samples containing six of the 10 possible combinations of the four beta-globin gene mutations under study, we were able to predict the mutations present in 23 out of 90 (26.4%) of the patients studied. These predictions were confirmed independently by the amplification refractory mutation system (ARMS) method. It is concluded that this non-radioactive PCR-SSCP method can be used to reliably identify mutations in beta-thalassemia patients, provided that suitable controls are available. However, usefulness of this method for determining the genotype of beta-thalassaemic individuals is obviously limited by the great number of controls required. Moreover, the ability to detect mutations by SSCP is in general lower compared to other methods, ARMS, DGGE or DHPLC, which are reported to detect 49.5% to 73% of the mutations present. The SSCP method is nevertheless much easier to employ than other methods and is especially successful for beta-thalassemia carriers. This method would thus be particularly useful for an initial screening of target groups (prenatal diagnosis)

  14. Protective effects of pulmonary epithelial lining fluid on oxidative stress and DNA single-strand breaks caused by ultrafine carbon black, ferrous sulphate and organic extract of diesel exhaust particles

    Energy Technology Data Exchange (ETDEWEB)

    Chuang, Hsiao-Chi [School of Respiratory Therapy, College of Medicine, Taipei Medical University, Taipei, Taiwan (China); Division of Pulmonary Medicine, Department of Internal Medicine, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan (China); Cheng, Yi-Ling; Lei, Yu-Chen [Institute of Occupational Medicine and Industrial Hygiene, College of Public Health, National Taiwan University, Taipei, Taiwan (China); Chang, Hui-Hsien [Institute of Environmental Health, College of Public Health, National Taiwan University, Taipei, Taiwan (China); Cheng, Tsun-Jen, E-mail: tcheng@ntu.edu.tw [Institute of Occupational Medicine and Industrial Hygiene, College of Public Health, National Taiwan University, Taipei, Taiwan (China); Department of Public Health, College of Public Health, National Taiwan University, Taipei, Taiwan (China)

    2013-02-01

    Pulmonary epithelial lining fluid (ELF) is the first substance to make contact with inhaled particulate matter (PM) and interacts chemically with PM components. The objective of this study was to determine the role of ELF in oxidative stress, DNA damage and the production of proinflammatory cytokines following physicochemical exposure to PM. Ultrafine carbon black (ufCB, 15 nm; a model carbonaceous core), ferrous sulphate (FeSO{sub 4}; a model transition metal) and a diesel exhaust particle (DEP) extract (a model organic compound) were used to examine the acellular oxidative potential of synthetic ELF and non-ELF systems. We compared the effects of exposure to ufCB, FeSO{sub 4} and DEP extract on human alveolar epithelial Type II (A549) cells to determine the levels of oxidative stress, DNA single-strand breaks and interleukin-8 (IL-8) production in ELF and non-ELF systems. The effects of ufCB and FeSO{sub 4} on the acellular oxidative potential, cellular oxidative stress and DNA single-strand breakage were mitigated significantly by the addition of ELF, whereas there was no decrease following treatment with the DEP extract. There was no significant effect on IL-8 production following exposure to samples that were suspended in ELF/non-ELF systems. The results of the present study indicate that ELF plays an important role in the initial defence against PM in the pulmonary environment. Experimental components, such as ufCB and FeSO{sub 4}, induced the production of oxidative stress and led to DNA single-strand breaks, which were moderately prevented by the addition of ELF. These findings suggest that ELF plays a protective role against PM-driven oxidative stress and DNA damage. -- Highlights: ► To determine the role of ELF in ROS, DNA damage and IL-8 after exposure to PM. ► ufCB, FeSO{sub 4} and DEP extract were used to examine the protective effects of ELF. ► PM-driven oxidative stress and DNA single-strand breakage were mitigated by ELF. ► The findings

  15. In vitro biocompatibility of nickel-titanium esthetic orthodontic archwires.

    Science.gov (United States)

    Rongo, Roberto; Valletta, Rosa; Bucci, Rosaria; Rivieccio, Virginia; Galeotti, Angela; Michelotti, Ambrosina; D'Antò, Vincenzo

    2016-09-01

    To investigate the cytotoxicity of nickel-titanium (NiTi) esthetic orthodontic archwires with different surface coatings. Three fully coated, tooth-colored NiTi wires (BioCosmetic, Titanol Cosmetic, EverWhite), two ion-implanted wires (TMA Purple, Sentalloy High Aesthetic), five uncoated NiTi wires (BioStarter, BioTorque, Titanol Superelastic, Memory Wire Superelastic, and Sentalloy), one β-titanium wire (TMA), and one stainless steel wire (Stainless Steel) were considered for this study. The wire samples were placed at 37°C in airtight test tubes containing Dulbecco's Modified Eagle's Medium (0.1 mg/mL) for 1, 7, 14, and 30 days. The cell viability of human gingival fibroblasts (HGFs) cultured with this medium was assessed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Data were analyzed by a two-way analysis of variance (α  =  .05). The highest cytotoxic effect was reached on day 30 for all samples. The archwires exhibited a cytotoxicity on HGFs ranging from "none" to "slight," with the exception of the BioTorque, which resulted in moderate cytotoxicity on day 30. Significant differences were found between esthetic archwires and their uncoated pairs only for BioCosmetic (P  =  .001) and EverWhite (P < .001). Under the experimental conditions, all of the NiTi esthetic archwires resulted in slight cytotoxicity, as did the respective uncoated wires. For this reason their clinical use may be considered to have similar risks to the uncoated archwires.

  16. An Ingested Orthodontic Wire Fragment: A Case Report.

    Science.gov (United States)

    Puryer, James; McNamara, Catherine; Sandy, Jonathan; Ireland, Tony

    2016-08-01

    Accidental ingestion or inhalation of foreign bodies has been widely documented, including incidents which occur whilst undertaking dental treatment. Most ingested objects pass through the gastrointestinal tract (GIT) spontaneously, but approximately 10%-20% need to be removed endoscopically and 1% require surgery. This case reports a complication arising from the accidental loss of an archwire fragment during maxillary archwire placement. It describes the immediate and subsequent management, including the use of radiographs to track the passage of the fragment through the gastro-intestinal tract. This case stresses the vigilance that dentists must take to prevent inhalation or ingestion of foreign bodies and the consequences of time-delays when management decisions are needed.

  17. Mechanical evaluation of space closure loops in Orthodontics

    OpenAIRE

    Rodrigues, Eduardo Uggeri; Maruo, Hiroshi; Guariza Filho, Odilon; Tanaka, Orlando; Camargo, Elisa Souza

    2011-01-01

    This study evaluated the mechanical performance of teardrop-shaped loops and teardrop-shaped loops with helix used in orthodontic space closure. Sixty retraction loops made with 0.019" x 0.025" stainless steel (SS) and beta-titanium (BT) wires were used. They were attached to a testing machine to measure the magnitudes of the sagittal force and the load-deflection ratio necessary for 1 mm, 2 mm and 3 mm activation. The results demonstrated that the BT alloy presented significantly smaller mea...

  18. Are claims made in orthodontic journal advertisements evidence-supported?

    NARCIS (Netherlands)

    Livas, Christos; Kouskoura, Thaleia; Ren, Yijin; Katsaros, Christos; Pandis, Nikolaos

    Objective: To examine the supporting evidence of advertisements published in six leading orthodontic journals. Materials and Methods: The 2012-2013 printed issues of American Journal of Orthodontics and Dentofacial Orthopedics, Australian Orthodontic Journal, Journal of Orthodontics, European

  19. Current advances in orthodontic pain

    Science.gov (United States)

    Long, Hu; Wang, Yan; Jian, Fan; Liao, Li-Na; Yang, Xin; Lai, Wen-Li

    2016-01-01

    Orthodontic pain is an inflammatory pain that is initiated by orthodontic force-induced vascular occlusion followed by a cascade of inflammatory responses, including vascular changes, the recruitment of inflammatory and immune cells, and the release of neurogenic and pro-inflammatory mediators. Ultimately, endogenous analgesic mechanisms check the inflammatory response and the sensation of pain subsides. The orthodontic pain signal, once received by periodontal sensory endings, reaches the sensory cortex for pain perception through three-order neurons: the trigeminal neuron at the trigeminal ganglia, the trigeminal nucleus caudalis at the medulla oblongata and the ventroposterior nucleus at the thalamus. Many brain areas participate in the emotion, cognition and memory of orthodontic pain, including the insular cortex, amygdala, hippocampus, locus coeruleus and hypothalamus. A built-in analgesic neural pathway—periaqueductal grey and dorsal raphe—has an important role in alleviating orthodontic pain. Currently, several treatment modalities have been applied for the relief of orthodontic pain, including pharmacological, mechanical and behavioural approaches and low-level laser therapy. The effectiveness of nonsteroidal anti-inflammatory drugs for pain relief has been validated, but its effects on tooth movement are controversial. However, more studies are needed to verify the effectiveness of other modalities. Furthermore, gene therapy is a novel, viable and promising modality for alleviating orthodontic pain in the future. PMID:27341389

  20. Effects of chlorhexidine (gel) application on bacterial levels and orthodontic brackets during orthodontic treatment.

    Science.gov (United States)

    Al-Bazi, Samar M; Abbassy, Mona A; Bakry, Ahmed S; Merdad, Leena A; Hassan, Ali H

    2016-01-01

    The objectives of this study were to evaluate the effects of applying 0.50% chlorhexidine (CHX) gel using the dental drug delivery system (3DS) on salivary Streptococcus mutans (S. mutans) and on the surface topography of metal and ceramic orthodontic brackets. The study involved 20 orthodontic patients with high levels of salivary S. mutans. The patients were treated with professional mechanical tooth cleaning followed by application of 0.50% CHX using individual trays (3DS). Salivary S. mutans levels were repeatedly measured 1, 2, 4, and 8 weeks post-treatment. In vitro study utilized forty ceramic and metallic brackets that were immersed in 0.50% CHX gel for 10 min, whereas another untreated forty brackets served as controls. The frictional resistances of stainless steel wires to the brackets before and after CHX treatment were recorded using a universal testing machine. Scanning electron microscopy was used to compare changes in the surface topography of brackets. Statistical analyses were used to determine the effect of CHX on bacterial count and to evaluate the effect of CHX on frictional resistance. According to the results of this study, S. mutans levels were reduced significantly (P brackets before or after application of CHX. (J Oral Sci 58, 35-42, 2016).

  1. A novel technique using DNA denaturation to detect multiply induced single-strand breaks in a hydrated plasmid DNA molecule by X-ray and 4He2+ ion irradiation

    International Nuclear Information System (INIS)

    Yokoya, A.; Shikazono, N.; Fujii, K.; Noguchi, M.; Urushibara, A.

    2011-01-01

    To detect multiple single-strand breaks (SSBs) produced in plasmid DNA molecules by direct energy deposition from radiation tracks, we have developed a novel technique using DNA denaturation by which irradiated DNA is analysed as single-strand DNA (SS-DNA). The multiple SSBs that arise in both strands of DNA, but do not induce a double-strand break, are quantified as loss of SS-DNA using agarose gel electrophoresis. We have applied this method to X-ray and 4 He 2+ ion-irradiated samples of fully hydrated pUC18 plasmid DNA. The fractions of both SS-DNA and closed circular DNA (CC-DNA) exponentially decrease with the increasing dose of X rays and 4 He 2+ ions. The efficiency of the loss of SS-DNA was half that of CC-DNA for both types of irradiation, indicating that one of two strands in DNA is not broken when one SSB is produced in CC-DNA by irradiation. Contrary to our initial expectation, these results indicate that SSBs are not multiply induced even by high linear energy transfer radiation distributed in both strands. (authors)

  2. The early UL31 gene of equine herpesvirus 1 encodes a single-stranded DNA-binding protein that has a nuclear localization signal sequence at the C-terminus

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Seongman; Chul Ahn, Byung; O' Callaghan, Dennis J. [Department of Microbiology and Immunology, Center for Molecular and Tumor Virology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932 (United States); Kim, Seong Kee, E-mail: skim1@lsuhsc.edu [Department of Microbiology and Immunology, Center for Molecular and Tumor Virology, Louisiana State University Health Sciences Center, Shreveport, LA 71130-3932 (United States)

    2012-10-25

    The amino acid sequence of the UL31 protein (UL31P) of equine herpesvirus 1 (EHV-1) has homology to that of the ICP8 of herpes simplex virus type 1 (HSV-1). Here we show that the UL31 gene is synergistically trans-activated by the IEP and the UL5P (EICP27). Detection of the UL31 RNA transcript and the UL31P in EHV-1-infected cells at 6 h post-infection (hpi) as well as metabolic inhibition assays indicated that UL31 is an early gene. The UL31P preferentially bound to single-stranded DNA over double-stranded DNA in gel shift assays. Subcellular localization of the green fluorescent protein (GFP)-UL31 fusion proteins revealed that the C-terminal 32 amino acid residues of the UL31P are responsible for the nuclear localization. These findings may contribute to defining the role of the UL31P single-stranded DNA-binding protein in EHV-1 DNA replication.

  3. The early UL31 gene of equine herpesvirus 1 encodes a single-stranded DNA-binding protein that has a nuclear localization signal sequence at the C-terminus

    International Nuclear Information System (INIS)

    Kim, Seongman; Chul Ahn, Byung; O’Callaghan, Dennis J.; Kim, Seong Kee

    2012-01-01

    The amino acid sequence of the UL31 protein (UL31P) of equine herpesvirus 1 (EHV-1) has homology to that of the ICP8 of herpes simplex virus type 1 (HSV-1). Here we show that the UL31 gene is synergistically trans-activated by the IEP and the UL5P (EICP27). Detection of the UL31 RNA transcript and the UL31P in EHV-1-infected cells at 6 h post-infection (hpi) as well as metabolic inhibition assays indicated that UL31 is an early gene. The UL31P preferentially bound to single-stranded DNA over double-stranded DNA in gel shift assays. Subcellular localization of the green fluorescent protein (GFP)–UL31 fusion proteins revealed that the C-terminal 32 amino acid residues of the UL31P are responsible for the nuclear localization. These findings may contribute to defining the role of the UL31P single-stranded DNA-binding protein in EHV-1 DNA replication.

  4. Induced magnetic moment in stainless steel components of orthodontic appliances in 1.5 T MRI scanners

    Science.gov (United States)

    Rollins, Nancy K.; Liang, Hui; Park, Yong Jong

    2015-01-01

    Purpose: Most orthodontic appliances are made of stainless steel materials and induce severe magnetic susceptibility artifacts in brain MRI. In an effort for correcting these artifacts, it is important to know the value of induced magnetic moments in all parts of orthodontic appliances. In this study, the induced magnetic moment of stainless steel orthodontic brackets, molar bands, and arch-wires from several vendors is measured. Methods: Individual stainless steel brackets, molar bands, and short segments of arch-wire were positioned in the center of spherical flask filled with water through a thin plastic rod. The induced magnetic moment at 1.5 T was determined by fitting the B0 map to the z-component of the magnetic dipole field using a computer routine. Results: The induced magnetic moment at 1.5 T was dominated by the longitudinal component mz, with a small contribution from the transverse components. The mz was insensitive to the orientation of the metal parts. The orthodontic brackets collectively dominated the magnetic dipole moment in orthodontic appliances. In brackets from six vendors, the total induced mz from 20 brackets for nonmolar teeth ranged from 0.108 to 0.158 (median 0.122) A ⋅ m2. The mz in eight molar bands with bracket attachment from two vendors ranged from 0.0004 to 0.0166 (median 0.0035) A ⋅ m2. Several full length arch wires had induced magnetic moment in the range of 0.006–0.025 (median 0.015) A ⋅ m2. Conclusions: Orthodontic brackets collectively contributed most to the total magnetic moment. Different types of brackets, molar bands, and arch wires all exhibit substantial variability in the induced magnetic moment. PMID:26429261

  5. Wire breakage in SLC wire profile monitors

    International Nuclear Information System (INIS)

    Field, C.; McCormick, D.; Raimondi, P.; Ross, M.

    1998-05-01

    Wire scanning beam profile monitors are used at the Stanford Linear Collider (SLC) for emittance preservation control and beam optics optimization. Twenty such scanners have proven most useful for this purpose and have performed a total of 1.5 million scans in the 4 to 6 years since their installation. Most of the essential scanners are equipped with 20 to 40 microm tungsten wires. SLC bunch intensities and sizes often exceed 2 x 10 7 particles/microm 2 (3C/m 2 ). The authors believe that this has caused a number of tungsten wire failures that appear at the ends of the wire, near the wire support points, after a few hundred scans are accumulated. Carbon fibers, also widely used at SLAC, have been substituted in several scanners and have performed well. In this paper, the authors present theories for the wire failure mechanism and techniques learned in reducing the failures

  6. Efficacy of protection by latex gloves during orthodontic therapy.

    Science.gov (United States)

    Doll, G M; Zentner, A; Balan, R; Sergl, H G

    2000-01-01

    The wearing of gloves during orthodontic or dental treatment is generally indicated for reasons of hygiene and protection against infection. This study was aimed at determining the extent and localization of perforations caused by the various orthodontic treatment techniques and interrupting the infection barrier. The impermeability was tested by means of a water retention test according to European standard EN 455, Part 1, performed on 1600 Centramed (Centramed, Koblenz), Tekmedic and SafeEx non-sterile disposable latex gloves (both by Safe Med, Switzerland) and Safe Gan latex gloves with an additional acrylate coating (also by Safe Med). The perforation rate in unused gloves was between 0.5% and 7.5%, rising on average to 11% with increasing use. 36% of the total number of lesions resulted from handling removable appliances, and 57% from handling fixed appliances, especially when replacing arch wires and elastics. Most lesions were in the thumb, index finger and palm region. Only 18% of the defects were noticed by the dentists themselves. The gloves worn by beginners in their first year of postgraduate orthodontic training had about twice as many defects as those worn by qualified orthodontists. When patients with an increased risk of infection are to be treated, additional hand disinfection measures should be taken and 2 pairs of gloves worn in view of the relatively unreliable protection offered by commercially available latex gloves.

  7. Quick response codes in Orthodontics

    Directory of Open Access Journals (Sweden)

    Moidin Shakil

    2015-01-01

    Full Text Available Quick response (QR code codes are two-dimensional barcodes, which encodes for a large amount of information. QR codes in Orthodontics are an innovative approach in which patient details, radiographic interpretation, and treatment plan can be encoded. Implementing QR code in Orthodontics will save time, reduces paperwork, and minimizes manual efforts in storage and retrieval of patient information during subsequent stages of treatment.

  8. Northcroft lecture: how has the spectrum of orthodontics changed over the past decades?

    Science.gov (United States)

    Melsen, Birte

    2011-06-01

    Three aspects have had a significant impact on orthodontics during the last few decades: the appliances being used, the anchorage being used and finally the distribution of patients being treated. Firstly, the marketing of appliances is increasingly leading the orthodontist to outsource important aspects of treatment such as wirebending and bracket positioning. Brackets and wires are being presented as the solution to all problems and metaphysical terms such as 'intelligent design,' 'working brackets' and 'intelligent wires' are dominating advertising and reducing the impact of evidence-based treatment approaches. Secondly, the introduction of skeletal anchorage has potentially widened the spectrum of orthodontics, allowing for treatments that could not be done with conventional appliances. Biomechanical knowledge is, however, mandatory if we agree that the system should not be abused. Thirdly, the orthodontic population comprises an increasing number of adult patients, many of whom are characterized by a degenerated dentition. The treatment of these patients requires a thorough knowledge not only of biomechanics but also of the reaction of the periodontal tissues to various types of loading. They can be treated only with custom-made appliances adapting the force systems and magnitude to the patient-specific treatment goal. In summary, the orthodontic world is being split between 'appliance-driven fast-food orthodontics' where the results to a large extent are dependent on both growth and function and 'orthodontist-driven' 'slow-food' treatments attempting to push the limits of the possible in relation to complicated problems and reversal of degeneration in adult patients. The latter treatments are performed with individualized appliances adapting the force system to the patient. This paper will attempt to summarize the bearing of these factors on present orthodontics.

  9. Antibacterial Activity of Orthodontic Cement Containing Quaternary Ammonium Polyethylenimine Nanoparticles Adjacent to Orthodontic Brackets

    OpenAIRE

    Eldad Sharon; Revital Sharabi; Adi Eden; Asher Zabrovsky; Gilad Ben-Gal; Esi Sharon; Yoav Pietrokovski; Yael Houri-Haddad; Nurit Beyth

    2018-01-01

    Enamel demineralization is a common problem found in patients using orthodontic devices, such as orthodontic braces. It was found that Streptoccocus mutans growth increases adjacent to orthodontic devices, which may result in caries development. Incorporated antibacterial quaternary ammonium polyethylenimine (QPEI) nanoparticles were previously shown to be highly efficacious against various bacteria. Combining antibacterial materials in orthodontic cement may be advantageous to prevent bacter...

  10. Torque differences due to the material variation of the orthodontic appliance: a finite element study.

    Science.gov (United States)

    Papageorgiou, Spyridon N; Keilig, Ludger; Vandevska-Radunovic, Vaska; Eliades, Theodore; Bourauel, Christoph

    2017-12-01

    Torque of the maxillary incisors is crucial to occlusal relationship and esthetics and can be influenced by many factors. The aim of this study was to assess the relative influence of the material of the orthodontic appliance (adhesive, bracket, ligature, and wire) on tooth displacements and developed stresses/strains after torque application. A three-dimensional upper right central incisor with its periodontal ligament (PDL) and alveolus was modeled. A 0.018-in. slot discovery® (Dentaurum, Ispringen, Germany) bracket with a rectangular 0.018 x 0.025-in. wire was generated. The orthodontic appliance varied in the material of its components: adhesive (composite resin or resin-modified glass ionomer cement), bracket (titanium, steel, or ceramic), wire (beta-titanium or steel), and ligature (elastomeric or steel). A total of 24 models were generated, and a palatal root torque of 5° was applied. Afterwards, crown and apex displacement, strains in the PDL, and stresses in the bracket were calculated and analyzed. The labial crown displacement and the palatal root displacement of the tooth were mainly influenced by the material of the wire (up to 150% variation), followed by the material of the bracket (up to 19% variation). The magnitude of strains developed in the PDL was primarily influenced by the material of the wire (up to 127% variation), followed by the material of the bracket (up to 30% variation) and the ligature (up to 13% variation). Finally, stresses developed at the bracket were mainly influenced by the material of the wire (up to 118% variation) and the bracket (up to 59% variation). The material properties of the orthodontic appliance and all its components should be considered during torque application. However, these in silico results need to be validated in vivo before they can be clinically extrapolated.

  11. Intracranial Migration of K-wire in a Bilateral Cleft Lip and Palate Patient

    Directory of Open Access Journals (Sweden)

    G Vimala

    2014-01-01

    A 16-year-old female patient with bilateral cleft lip and palate reported for orthodontic treatment. Upon routine investigations, presence of a K-wire that was fixed when patient was around 6 months old, and now had migrated intracranially was made. As K-wire is notorious for its migration to distant places and, that any further impending migration, may in future present a serious problem, the wire was removed after necessary precautions. This case report should be an eye opener and post a caution to general, medical, surgical practitioners and orthodontists.

  12. An evaluation of two types of nickel-titanium wires in terms of micromorphology and nickel ions' release following oral environment exposure.

    Science.gov (United States)

    Ghazal, Abdul Razzak A; Hajeer, Mohammad Y; Al-Sabbagh, Rabab; Alghoraibi, Ibrahim; Aldiry, Ahmad

    2015-01-01

    This study aimed to compare superelastic and heat-activated nickel-titanium orthodontic wires' surface morphology and potential release of nickel ions following exposure to oral environment conditions. Twenty-four 20-mm-length distal cuts of superelastic (NiTi Force I®) and 24 20-mm-length distal cuts of heat-activated (Therma-Ti Lite®) nickel-titanium wires (American Orthodontics, Sheboygan, WI, USA) were divided into two equal groups: 12 wire segments left unused and 12 segments passively exposed to oral environment for 1 month. Scanning electron microscopy and atomic force microscopy were used to analyze surface morphology of the wires which were then immersed in artificial saliva for 1 month to determine potential nickel ions' release by means of atomic absorption spectrophotometer. Heat-activated nickel-titanium (NiTi) wires were rougher than superelastic wires, and both types of wires released almost the same amount of Ni ions. After clinical exposure, more surface roughness was recorded for superelastic NiTi wires and heat-activated NiTi wires. However, retrieved superelastic NiTi wires released less Ni ions in artificial saliva after clinical exposure, and the same result was recorded regarding heat-activated wires. Both types of NiTi wires were obviously affected by oral environment conditions; their surface roughness significantly increased while the amount of the released Ni ions significantly declined.

  13. Base Information Transport Infrastructure Wired (BITI Wired)

    Science.gov (United States)

    2016-03-01

    2016 Major Automated Information System Annual Report Base Information Transport Infrastructure Wired (BITI Wired) Defense Acquisition Management...Combat Information Transport System program was restructured into two pre-Major Automated Information System (pre-MAIS) components: Information...Major Automated Information System MAIS OE - MAIS Original Estimate MAR – MAIS Annual Report MDA - Milestone Decision Authority MDD - Materiel

  14. A Clinical Assessment of Corticotomy Facilitated Orthodontics in the Retraction of Maxillary Anterior Segment

    Directory of Open Access Journals (Sweden)

    Chagam Manjunatha Reddy

    2014-01-01

    Full Text Available Corticotomy facilitated orthodontics has been practising for longer periods, but a novel approach of a periapical incision to make horizontal cuts 5 mm above the apex of the maxillary anteriors along with vertical cuts requires a detailed investigation. The purpose of this study was to assess the outcome of skeletal cases with spacing in the anteriors using a novel method of modified surgical approach utilizing the corticotomy facilitated orthodontics. Patients for this study were randomly selected from a very intensive screening in the Department of Orthodontics, Saveetha Dental College and Hospital, Chennai, out of which seven patients were selected for this procedure. The patients were subjected for corticotomy procedure under local anesthesia after the leveling and aligning stage (0.019" × 0.025" stainless steel wire, and the retraction was continued still the space was closed. AutoCAD and palatal rugae assessment programs were used to assess the orthodontic tooth movement. The results of the study demonstrated that when compared to conventional orthodontics alone, the corticotomy facilitated approach produced faster tooth movements in all the seven patients reducing the overall treatment time by 60%. The use of this modified technique of labial, palatal and apical cuts could be beneficial for adult patients with difficult surgical access from the lingual side.

  15. DNA damage in oral mucosa cells of patients with fixed orthodontic appliances.

    Directory of Open Access Journals (Sweden)

    Farzin Heravi

    2013-12-01

    Full Text Available The release of toxic metal ions from orthodontic alloys has induced concerns regarding the biocompatibility of fixed appliances. This study investigated the genotoxic effect of metal appliances in a sample of patients undergoing fixed orthodontic treatment.The study included twenty-five healthy individuals requiring orthodontic therapy in both jaws. The patients were treated by stainless steel orthodontic brackets and nickel-titanium or stainless steel arch wires. The oral mucosa cells were gathered just before the appliance placement and 9 months later. The cells were centrifuged, fixed and dropped onto slides. After staining, the micronucleus (MN assay was used to determine genome alteration. The data were analyzed by paired sample t-test.The mean micronuclei frequency in the buccal mucosa was 10.6 ± 5.7 per 1000 cells before the appliance placement and 9.2 ± 6.37 per 1000 cells 9 months later. No significant difference was found in the MN count before and 9 months after therapy (p=0.336.Under the conditions used in this study, application of fixed orthodontic appliances did not expose healthy individuals to increased risk of DNA damage in oral mucosa cells.

  16. Wire bonding in microelectronics

    CERN Document Server

    Harman, George G

    2010-01-01

    Wire Bonding in Microelectronics, Third Edition, has been thoroughly revised to help you meet the challenges of today's small-scale and fine-pitch microelectronics. This authoritative guide covers every aspect of designing, manufacturing, and evaluating wire bonds engineered with cutting-edge techniques. In addition to gaining a full grasp of bonding technology, you'll learn how to create reliable bonds at exceedingly high yields, test wire bonds, solve common bonding problems, implement molecular cleaning methods, and much more. Coverage includes: Ultrasonic bonding systems and technologies, including high-frequency systems Bonding wire metallurgy and characteristics, including copper wire Wire bond testing Gold-aluminum intermetallic compounds and other interface reactions Gold and nickel-based bond pad plating materials and problems Cleaning to improve bondability and reliability Mechanical problems in wire bonding High-yield, fine-pitch, specialized-looping, soft-substrate, and extreme-temperature wire bo...

  17. French Board of Orthodontics.

    Science.gov (United States)

    De Sousa Beltrao, Paulo Augusto

    2010-09-01

    A. February 2001 (15.11 y). Molar Class I, skeletal Class I, normodivergent, anterior deficiency: -17 mm, bimaxillary protrusion. Preparation of arches (alignment, levelling, canine retraction). Correction of arches (space closure). Occlusal finishing (artistic bends). B. POSTTREATMENT RECORDS: February 2003 (17.11 y). Maxillary Hawley plate for nighttime use and bonded lingual wire from 33 to 43. C. POSTRETENTION RECORDS: December 2003 (18.9 y). Copyright © 2010 CEO. Published by Elsevier Masson SAS. All rights reserved.

  18. Gingival Crevicular Fluid Turnover Markers in Premenopausal vs Postmenopausal Women receiving Orthodontic Treatment.

    Science.gov (United States)

    Bitra, Anusha; Rani, B Jhansi; Agarkar, Sanket S; Parihar, Anuj S; Vynath, Gopinath P; Grover, Shekhar

    2017-10-01

    Orthodontic treatment is one of the commonly used dental treatments. Orthodontic forces act on the bone by modulating the biomolecules, chiefly the osteoprotegerin (OPG), osteopontin (OPN), receptor activator of nuclear factor kappa-B (RANK), and RANK ligand (RANKL) (OPG ligand). Hormonal changes are known to cause marked alteration in the levels of these biomolecules. Hence, we planned this study to evaluate the response of bone biomarkers in the gingival crevicular fluid (GCF) in postmenopausal women undergoing fixed orthodontic therapy. This study included assessment of 50 subjects who underwent orthodontic treatment from June 2012 to July 2016. All the patients were divided into two study groups with 25 patients in each group: premenopausal group and postmenopausal group. Similar orthodontic wires were used for controlling the forces applied in subjects of both the study groups and their GCF levels of RANKL, and OPN was assessed at baseline and 24 hours after the activation of orthodontic forces. All the results were compiled, assessed, and analyzed by Statistical Package for the Social Sciences software version 16.0. Chi-square test, Student's t-test, and Mann-Whitney U test were used for the assessment of the level of significance. The mean values of RANKL and OPN in the premenopausal and postmenopausal groups were found to be 241.52 and 317.15 pg/μL respectively. The mean values of RANKL at baseline in the premenopausal and postmenopausal groups were found to be 7.15 and 3.84 pg/μL respectively. Nonsignificant results were obtained while comparing mean OPN and RANKL level alteration in between the two study groups. The mean alterations in the GCF levels of bone biomarkers are similar for both premenopausal and postmeno-pausal women. For women with either premenopausal or postmenopausal status, orthodontic treatment appears to be equally safer.

  19. Reparation in unicellular green algae during chronic exposure to the action of mutagenic factors. II. Restoration of single-stranded DNA breaks following exposure of Chlamydomonas reinchardii to gamma-irradiation

    International Nuclear Information System (INIS)

    Sergeeva, S.A.; Ptitsina, S.N.; Shevchenko, V.A.

    1986-01-01

    The restoration of single-stranded breaks in the DNA in different strains of unicellular green algae (chlamydomonads) during chronic exposure to the action of mutagenic factors following γ-irradiation was investigated. It was shown that the restoration of DNA breaks was most effective in the case of strain M γ/sup mt + /, which is resistant to radiation. Strains, that were sensitive to UV irradiation showed a similar order of DNA break restoration as the wild-type strain. Strain UVS-1 showed a higher level of restoration than the wild-type strain. The data indicated that chlamydomonads have different pathways of reparation, which lead to the restoration of breaks induced by γ-irradiation and UV-rays

  20. C-terminal phenylalanine of bacteriophage T7 single-stranded DNA-binding protein is essential for strand displacement synthesis by T7 DNA polymerase at a nick in DNA.

    Science.gov (United States)

    Ghosh, Sharmistha; Marintcheva, Boriana; Takahashi, Masateru; Richardson, Charles C

    2009-10-30

    Single-stranded DNA-binding protein (gp2.5), encoded by gene 2.5 of bacteriophage T7, plays an essential role in DNA replication. Not only does it remove impediments of secondary structure in the DNA, it also modulates the activities of the other replication proteins. The acidic C-terminal tail of gp2.5, bearing a C-terminal phenylalanine, physically and functionally interacts with the helicase and DNA polymerase. Deletion of the phenylalanine or substitution with a nonaromatic amino acid gives rise to a dominant lethal phenotype, and the altered gp2.5 has reduced affinity for T7 DNA polymerase. Suppressors of the dominant lethal phenotype have led to the identification of mutations in gene 5 that encodes the T7 DNA polymerase. The altered residues in the polymerase are solvent-exposed and lie in regions that are adjacent to the bound DNA. gp2.5 lacking the C-terminal phenylalanine has a lower affinity for gp5-thioredoxin relative to the wild-type gp2.5, and this affinity is partially restored by the suppressor mutations in DNA polymerase. gp2.5 enables T7 DNA polymerase to catalyze strand displacement DNA synthesis at a nick in DNA. The resulting 5'-single-stranded DNA tail provides a loading site for T7 DNA helicase. gp2.5 lacking the C-terminal phenylalanine does not support this event with wild-type DNA polymerase but does to a limited extent with T7 DNA polymerase harboring the suppressor mutations.

  1. C-terminal Phenylalanine of Bacteriophage T7 Single-stranded DNA-binding Protein Is Essential for Strand Displacement Synthesis by T7 DNA Polymerase at a Nick in DNA*

    Science.gov (United States)

    Ghosh, Sharmistha; Marintcheva, Boriana; Takahashi, Masateru; Richardson, Charles C.

    2009-01-01

    Single-stranded DNA-binding protein (gp2.5), encoded by gene 2.5 of bacteriophage T7, plays an essential role in DNA replication. Not only does it remove impediments of secondary structure in the DNA, it also modulates the activities of the other replication proteins. The acidic C-terminal tail of gp2.5, bearing a C-terminal phenylalanine, physically and functionally interacts with the helicase and DNA polymerase. Deletion of the phenylalanine or substitution with a nonaromatic amino acid gives rise to a dominant lethal phenotype, and the altered gp2.5 has reduced affinity for T7 DNA polymerase. Suppressors of the dominant lethal phenotype have led to the identification of mutations in gene 5 that encodes the T7 DNA polymerase. The altered residues in the polymerase are solvent-exposed and lie in regions that are adjacent to the bound DNA. gp2.5 lacking the C-terminal phenylalanine has a lower affinity for gp5-thioredoxin relative to the wild-type gp2.5, and this affinity is partially restored by the suppressor mutations in DNA polymerase. gp2.5 enables T7 DNA polymerase to catalyze strand displacement DNA synthesis at a nick in DNA. The resulting 5′-single-stranded DNA tail provides a loading site for T7 DNA helicase. gp2.5 lacking the C-terminal phenylalanine does not support this event with wild-type DNA polymerase but does to a limited extent with T7 DNA polymerase harboring the suppressor mutations. PMID:19726688

  2. Water Desalination with Wires

    NARCIS (Netherlands)

    Porada, S.; Sales, B.B.; Hamelers, H.V.M.; Biesheuvel, P.M.

    2012-01-01

    We show the significant potential of water desalination using a novel capacitive wire-based technology in which anode/cathode wire pairs are constructed from coating a thin porous carbon electrode layer on top of electrically conducting rods (or wires). By alternately dipping an array of electrode

  3. Effect of wire size on maxillary arch force/couple systems for a simulated high canine malocclusion.

    Science.gov (United States)

    Major, Paul W; Toogood, Roger W; Badawi, Hisham M; Carey, Jason P; Seru, Surbhi

    2014-12-01

    To better understand the effects of copper nickel titanium (CuNiTi) archwire size on bracket-archwire mechanics through the analysis of force/couple distributions along the maxillary arch. The hypothesis is that wire size is linearly related to the forces and moments produced along the arch. An Orthodontic Simulator was utilized to study a simplified high canine malocclusion. Force/couple distributions produced by passive and elastic ligation using two wire sizes (Damon 0.014 and 0.018 inch) measured with a sample size of 144. The distribution and variation in force/couple loading around the arch is a complicated function of wire size. The use of a thicker wire increases the force/couple magnitudes regardless of ligation method. Owing to the non-linear material behaviour of CuNiTi, this increase is less than would occur based on linear theory as would apply for stainless steel wires. The results demonstrate that an increase in wire size does not result in a proportional increase of applied force/moment. This discrepancy is explained in terms of the non-linear properties of CuNiTi wires. This non-proportional force response in relation to increased wire size warrants careful consideration when selecting wires in a clinical setting. © 2014 British Orthodontic Society.

  4. Corticotomy-assisted orthodontic treatment

    Science.gov (United States)

    Hassan, Ali H.; Al-Saeed, Samar H.; Al-Maghlouth, Basma A.; Bahammam, Maha A.; Linjawi, Amal I.; El-Bialy, Tarek H.

    2015-01-01

    Objectives: To systematically review the literature to assess the quality of evidence related to corticotomy-assisted orthodontic treatment (CAOT) as adjunctive treatment in orthodontics. Methods: The study was conducted in the Faculty of Dentistry, King Abdulaziz University, Jeddah, Saudi Arabia between 2013 and 2014. Various electronic databases were searched and abstracts were retrieved. Defined inclusion criteria were then applied to the obtained original articles for further evaluation by 2 examiners independently. The criteria of selection included human, or animal studies, which assessed some aspects of CAOT and/or the biological principles behind it. Case reports and series were excluded. The quality of the studies was evaluated by the methodological score for clinical trials developed. Results: Fourteen articles were retrieved initially, but only 12 articles were finally selected for the study. The CAOT was found to accelerate tooth movement by 2-2.5 folds when compared with conventional orthodontic tooth movement. The CAOT was found safe on periodontal health and exhibits no or little risk of root resorption. A localized turnover of alveolar spongiosa and the absence of a hyalinized zone was the acceptable biological explanation of CAOT. There is no evidence to support that CAOT enhances the movement of ankylosed teeth, closing old extraction sites, post-orthodontic stability, or transverse expansion. Conclusions: Corticotomy-assisted orthodontic treatment should be considered with caution. Long term randomized clinical trials are still needed. PMID:26108582

  5. [Root resorption and orthodontic treatment].

    Science.gov (United States)

    Sebbar, M; Bourzgui, F

    2011-09-01

    The aim of our study was to investigate the prevalence of root resorption during and at the end of orthodontic treatment and to assess its relationship with age, sex and treatment with or without extractions. Our study included 82 patients (51 women and 31 men) aged between 6 and 38 years, who received orthodontic treatment. Evaluation of root resorption was performed on panoramics at the beginning and at the end of orthodontic treatment. All the teeth were observed. The degree of root resorption was increased respectively by the standards in four ordinal levels (4). Data analysis was performed by Epi Info 6.0. Root resorption was present in all the teeth and maxillary incisors are the most affected. The correlation between age and root resorption was significant (p = 0.008). Women were more affected by resorption (P = 0.002). Patients treated with extraction showed more root resorption (p = 0.12). Our results suggest that orthodontic treatment is involved in the development of root resorption. The most often teeth resorbed are maxillary incisors. Age, sex and orthodontic extractions can be considered as risk factors for root resorption.

  6. PS wire chamber

    CERN Multimedia

    1970-01-01

    A wire chamber used at CERN's Proton Synchrotron accelerator in the 1970s. Multi-wire detectors contain layers of positively and negatively charged wires enclosed in a chamber full of gas. A charged particle passing through the chamber knocks negatively charged electrons out of atoms in the gas, leaving behind positive ions. The electrons are pulled towards the positively charged wires. They collide with other atoms on the way, producing an avalanche of electrons and ions. The movement of these electrons and ions induces an electric pulse in the wires which is collected by fast electronics. The size of the pulse is proportional to the energy loss of the original particle.

  7. Orthodontic parotitis: a rare complication from an orthodontic appliance.

    Science.gov (United States)

    McCarthy, Eileen; Cobb, Alistair R M

    2012-12-01

    A case is presented of a 14-year-old female undergoing orthodontic fixed appliance treatment who presented with right facial swelling in the parotid region. An initial diagnosis of acute infective parotitis was made by her primary care clinician. However, after clinical examination and ultrasonographic imaging, a diagnosis of salivary stasis secondary to inflammatory occlusion of Stensen's ductal orifice was made. The ductal orifice had been traumatized by the adjacent orthodontic appliance. This has not been described before in the literature. The differential diagnosis of parotid enlargement in children is discussed.

  8. Effect of wire shape on wire array discharge

    Energy Technology Data Exchange (ETDEWEB)

    Shimomura, N.; Tanaka, Y.; Yushita, Y.; Nagata, M. [University of Tokushima, Department of Electrical and Electronic Engineering, Tokushima (Japan); Teramoto, Y.; Katsuki, S.; Akiyama, H. [Kumamoto University, Department of Electrical and Computer Engineering, Kumamoto (Japan)

    2001-09-01

    Although considerable investigations have been reported on z-pinches to achieve nuclear fusion, little attention has been given from the point of view of how a wire array consisting of many parallel wires explodes. Instability existing in the wire array discharge has been shown. In this paper, the effect of wire shape in the wire array on unstable behavior of the wire array discharge is represented by numerical analysis. The claws on the wire formed in installation of wire may cause uniform current distribution on wire array. The effect of error of wire diameter in production is computed by Monte Carlo Method. (author)

  9. Effect of wire shape on wire array discharge

    International Nuclear Information System (INIS)

    Shimomura, N.; Tanaka, Y.; Yushita, Y.; Nagata, M.; Teramoto, Y.; Katsuki, S.; Akiyama, H.

    2001-01-01

    Although considerable investigations have been reported on z-pinches to achieve nuclear fusion, little attention has been given from the point of view of how a wire array consisting of many parallel wires explodes. Instability existing in the wire array discharge has been shown. In this paper, the effect of wire shape in the wire array on unstable behavior of the wire array discharge is represented by numerical analysis. The claws on the wire formed in installation of wire may cause uniform current distribution on wire array. The effect of error of wire diameter in production is computed by Monte Carlo Method. (author)

  10. Tensile test and interface retention forces between wires and composites in lingual fixed retainers.

    Science.gov (United States)

    Paolone, Maria Giacinta; Kaitsas, Roberto; Obach, Patricia; Kaitsas, Vasilios; Benedicenti, Stefano; Sorrenti, Eugenio; Barberi, Fabrizio

    2015-06-01

    In daily orthodontic clinical practice retention is very important, and lingual retainers are part of this challenge. The failure of lingual retainers may be due to many factors. The aim of this study was to assess the retention forces and mechanical behavior of different types of wires matched with different kinds of composites in lingual retainers. A tensile test was performed on cylindrical composite test specimens bonded to orthodontic wires. The specimens were constructed using four different wires: a straight wire (Remanium .016×.022″ Dentaurum), two round twisted wires (Penta One .0215″ Masel, Gold Penta Twisted .0215″ Gold N'braces) and a rectangular braided wire (D-Rect .016×.022″ Ormco); and three composites: two micro-hybrids (Micro-Hybrid Enamel Plus HFO Micerium, and Micro-Hybrid SDR U Dentsply) and a micro-nano-filled composite (Micro-Nano-Filled Transbond LR 3M). The test was performed at a speed of 10mm/min on an Inström device. The wire was fixed with a clamp. The results showed that the bonding between wires and composites in lingual fixed retainers seemed to be lowest for rectangular smooth wires and increased in round twisted and rectangular twisted wires where the bonding was so strong that the maximum tension/bond strength was greater than the ultimate tensile strength of the wire. The highest values were in rectangular twisted wires. Concerning the composites, hybrid composites had the lowest interface bonding values and broke very quickly, while the nano- and micro-composites tolerated stronger forces and displayed higher bonding values. The best results were observed with the golden twisted wire and reached 21.46 MPa with the Transbond composite. With the rectangular braided wire the retention forces were so high that the Enamel Plus composite fractured when the load exceeded 154.6 N/MPa. When the same wire was combined with the Transbond LR either the wire or the composite broke when the force exceeded 240 N. The results of this

  11. The World of Orthodontic apps.

    Directory of Open Access Journals (Sweden)

    Gaurav Gupta

    2017-01-01

    Full Text Available The usage of the portable electronic devices such as the smartphones and handheld tablets has increased over the years, and this is true in the health-care industry also. This is because of the development of various patient management softwares. The use of apps to manage, educate, and inform patient is not uncommon among orthodontists nowadays. The aim of this article was to review the various apps available on the Google Play Store and iOS Apple Store for orthodontists and patients. Four smartphones using orthodontically relevant keywords such as orthodontics, orthodontists, and braces were searched and reviewed in detail. Out of the 354 orthodontically relevant apps available in both Android and Apple operating systems, the apps could be categorized as orthodontist-related apps or patient-related apps. Under these categories they could be further classified as practice managements apps, patient education apps, model analysis apps, tooth material calculators, patient reminder apps, etc.

  12. Metal Hypersensitivity in Orthodontic Patients

    Directory of Open Access Journals (Sweden)

    Sandhya Maheshwari Sanjeev K

    2015-06-01

    Full Text Available Orthodontic treatment of individuals with metal hypersensitivity is a matter of concern for the orthodontist. Orthodontic appliances contain metals like Nickel, Cobalt and Chromium etc. Metals may cause allergic reactions and are known as allergens. Reaction to these metals is due to biodegradation of metals in the oral cavity. This may lead to the formation of corrosion products and their exposure to the patient. Nickel is the most common metal to cause hypersensitivity reaction. Chromium ranks second among the metals, known to trigger allergic reactions. The adverse biological reactions to these metals may include hypersensitivity, dermatitis and asthma. In addition, a significant carcinogenic and mutagenic potential has been demonstrated. The orthodontist must be familiar with the best possible alternative treatment modalities to provide the safest, most effective care possible in these cases. The present article focuses on the issue of metal hypersensitivity and its management in orthodontic

  13. Myths and realities in orthodontics.

    Science.gov (United States)

    Fleming, P S; Springate, S D; Chate, R A C

    2015-02-16

    Comprehensive orthodontic treatment typically comprises an initial phase of alignment over a period of four to six months, followed by vertical, transverse and antero-posterior corrections, space closure, finishing and detailing to enhance dental and facial aesthetics and function. Each course of treatment involves a series of decisions and alternatives relating to objectives, appliance design and treatment mechanics. In recent years there has been increasing interest in short-term approaches to treatment with more limited objectives and the avoidance of phases traditionally considered integral to successful treatment. In this review the veracity of accepted truths in orthodontics are discussed; specifically, the importance of initial molar relationship, final incisor relationship, the merits of orthodontic extractions, anticipated treatment times, the value of modern fixed appliance systems, the importance of torque expression and the relative merits of bonded retainers and inter-proximal reduction are considered.

  14. Effect of load deflection on corrosion behavior of NiTi wire.

    Science.gov (United States)

    Liu, I H; Lee, T M; Chang, C Y; Liu, C K

    2007-06-01

    For dental orthodontic applications, NiTi wires are used under bending conditions in the oral environment for a long period. The purpose of this study was to investigate the effect of bending stress on the corrosion of NiTi wires using potentiodynamic and potentiostatic tests in artificial saliva. The results indicated that bending stress induces a higher corrosion rate of NiTi wires in passive regions. It is suggested that the passive oxide film of specimens would be damaged under bending conditions. Auger electron spectroscopic analysis showed a lower thickness of passive films on stressed NiTi wires compared with unstressed specimens in the passive region. By scanning electron microscopy, localized corrosion was observed on stressed Sentalloy specimens after a potentiodynamic test at pH 2. In conclusion, this study indicated that bending stress changed the corrosion properties and surface characteristics of NiTi wires in a simulated intra-oral environment.

  15. Orthodontics is temporomandibular disorder-neutral.

    Science.gov (United States)

    Manfredini, Daniele; Stellini, Edoardo; Gracco, Antonio; Lombardo, Luca; Nardini, Luca Guarda; Siciliani, Giuseppe

    2016-07-01

    To assess if subjects with a clinical diagnosis of temporomandibular disorders (TMDs) have a similar prevalence of orthodontic history as a population of TMD-free individuals and to assess if those subjects who have a history of ideal orthodontics have fewer symptoms than those with a history of nonideal orthodontics. Two groups of age- and sex-matched individuals belonging to either a study ("TMD") or a control group were recruited. Subjects who underwent orthodontic treatment were classified as having a history of ideal or nonideal orthodontics based on the current presence of normal values in five reference occlusal features. The correlation with a history of orthodontic treatment was not clinically significant for any of the TMD diagnoses (ie, muscle pain, joint pain, disc displacement, arthrosis), with Phi (Φ) coefficient values within the -0.120 to 0.058 range. Within the subset of patients with a history of orthodontics, the correlation of ideal or nonideal orthodontic treatment with TMD diagnoses was, in general, not clinically relevant or was weakly relevant. Findings confirmed the substantial absence of clinically significant effects of orthodontics as far as TMD is concerned. The very low correlation values of a negative or positive history of ideal or nonideal orthodontics with the different TMD diagnoses suggest that orthodontic treatment could not have a true role for TMD.

  16. Periodontal status of Pakistani orthodontic patients

    Directory of Open Access Journals (Sweden)

    Yousuf MOOSA

    2015-01-01

    Full Text Available The objective of this study was to evaluate and compare the periodontal status of orthodontic patients and non-orthodontic patients, aged 15–28 years, of both genders. The cross-sectional study included 100 orthodontic and 100 non-orthodontic patients evaluated using a Community Periodontal Index for Treatment Need (CPITN probe on the index teeth. A questionnaire was distributed to the participants to assess and evaluate the use of oral hygiene aids. The data were analyzed using SPSS version 17, and various comparisons were performed using the chi-square test. The study revealed that there was a statistically significant association in CPITN scores between the orthodontic and non-orthodontic patients (p < 0.01. The study showed that patients undergoing orthodontic treatment have increased plaque accumulation and probing depth resulting in periodontal tissue destruction. Proper oral hygiene practices and interdental aids should be employed to control plaque.

  17. An update on periodontic-orthodontic interrelationships.

    Science.gov (United States)

    Dannan, Aous

    2010-01-01

    Talking about periodontic-orthodontic interrelationships is related primarily to the 1960s, where a generalized increase in salivary bacterial counts, especially Lactobacillus, had been shown after orthodontic band placement. The purpose of this article is to provide the dental practitioner with basic understanding of the interrelationship between periodontics and orthodontics by means of representing classical studies, and, to give an update on this topic by demonstrating the most recent opinions concerning periodontic-orthodontic interrelationships. Specific areas reviewed are the ability of orthodontic treatment to afford some degree of protection against periodontal breakdown, short-term and long-term effects of orthodontic treatment on the periodontium, and some mucogingival considerations. Topics considering orthodontic treatment in periodontally compromised patients were not included in this review. While past studies have shown that orthodontic treatment can positively affect the periodontal health, recent reviews indicate an absence of reliable evidence for the positive effects of orthodontic therapy on patients' periodontal status. Periodontic-orthodontic interrelationships are still controversial issues. However, a standard language between the periodontist and the orthodontist must always be established to eliminate the existing communications barrier, and to improve the outcomes of the whole treatment.

  18. An update on periodontic-orthodontic interrelationships

    Directory of Open Access Journals (Sweden)

    Dannan Aous

    2010-01-01

    Full Text Available Talking about periodontic-orthodontic interrelationships is related primarily to the 1960s, where a generalized increase in salivary bacterial counts, especially Lactobacillus, had been shown after orthodontic band placement. The purpose of this article is to provide the dental practitioner with basic understanding of the interrelationship between periodontics and orthodontics by means of representing classical studies, and, to give an update on this topic by demonstrating the most recent opinions concerning periodontic-orthodontic interrelationships. Specific areas reviewed are the ability of orthodontic treatment to afford some degree of protection against periodontal breakdown, short-term and long-term effects of orthodontic treatment on the periodontium, and some mucogingival considerations. Topics considering orthodontic treatment in periodontally compromised patients were not included in this review. While past studies have shown that orthodontic treatment can positively affect the periodontal health, recent reviews indicate an absence of reliable evidence for the positive effects of orthodontic therapy on patients′ periodontal status. Periodontic-orthodontic interrelationships are still controversial issues. However, a standard language between the periodontist and the orthodontist must always be established to eliminate the existing communications barrier, and to improve the outcomes of the whole treatment.

  19. Patients' and parents' expectations of orthodontic treatment.

    Science.gov (United States)

    Hiemstra, Renske; Bos, Annemieke; Hoogstraten, Johan

    2009-12-01

    To investigate the expectations of children and their primary care-givers towards orthodontic treatment and to compare the results with those of a UK sample. A questionnaire survey of children and their primary care-givers attending for their first consultation. The Department of Orthodontics at the Academic Centre for Dentistry Amsterdam (ACTA), the Netherlands. A total of 168 subjects (84 patients and 84 parents) completed the questionnaire. The children were aged 10 to 14 years. The responses of the children and parents and differences between boys and girls were examined using parametric statistical methods. The data from the Dutch sample were compared with a similar UK sample. Patients and parents shared similar expectations of orthodontic treatment, with the exception of expectations of having a brace fitted at the first appointment, orthodontic treatment involving headgear, any problems with orthodontic treatment, duration of orthodontic treatment and concerning reactions from the public. Among the child participants, boys and girls only differed in their expectations of orthodontic treatment involving jaw surgery. Differences between Dutch and English participants were found regarding the first visit, type of orthodontic treatment, reactions from the public, and pain and problems with orthodontic treatment. Since the expectations of patients and their parents differ on several aspects, effective communication between the orthodontist, patient and parent is considered to be essential. Our hypothesis that Dutch patients' and parents' expectations of orthodontic treatment differ from the expectations of English patients and parents was supported.

  20. Evaluation of scalp hair nickel and chromium level changes in patients with fixed orthodontic appliance: a one-year follow-up study.

    Science.gov (United States)

    Jamshidi, Saman; Rahmati Kamel, Manouchehr; Mirzaie, Maysam; Sarrafan, Ahmadreza; Khafri, Soraya; Parsian, Hadi

    2018-01-01

    The release of metal ions from orthodontic appliances is part of the dissolution and biomechanical processes of alloys. Nickel (Ni) and chromium (Cr) are the elements commonly used in the manufacture of various components of fixed orthodontic appliances, including bands, brackets and wires. This study was aimed to measure the Ni and Cr ions levels in the scalp hair of patients treated with fixed orthodontic appliances in comparison of the control group. The patient group consisted of 24 patients treated with fixed orthodontic appliances for one year, while the control group included 28 healthy individuals without orthodontic appliances. Analysis of the Cr and Ni was performed using atomic absorption spectrophotometer by graphite furnace method. The data were analyzed via student and paired samples t-test and ANOVA repeated measurement test. After one year, the levels of Ni and Cr in two groups showed significant differences (0.086 ± 0.007 and 0.258 ± 0.009 µg/g for control group and 0.149 ± 0.010 and 0.339 ± 0.013 µg/g for patient group, respectively for Ni and Cr, p orthodontic appliances and considering the cytotoxic and allergic effects of these ions, changing the ingredients in fixed orthodontic appliances is suggested for the future.

  1. Towards plant wires

    OpenAIRE

    Adamatzky, Andrew

    2014-01-01

    In experimental laboratory studies we evaluate a possibility of making electrical wires from living plants. In scoping experiments we use lettuce seedlings as a prototype model of a plant wire. We approximate an electrical potential transfer function by applying direct current voltage to the lettuce seedlings and recording output voltage. We analyse oscillation frequencies of the output potential and assess noise immunity of the plant wires. Our findings will be used in future designs of self...

  2. Correlation between cell survival and DNA single-strand break repair proficiency in the Chinese hamster ovary cell lines AA8 and EM9 irradiated with 365-nm ultraviolet-A radiation

    Energy Technology Data Exchange (ETDEWEB)

    Churchill, M.E.; Peak, J.G.; Peak, M.J. (Argonne National Lab., IL (USA))

    1991-02-01

    Cell survival parameters and the induction and repair of DNA single-strand breaks were measured in two Chinese hamster ovary cell lines after irradiation with monochromatic UVA radiation of wavelength 365 nm. The radiosensitive mutant cell line EM9 is known to repair ionizing-radiation-induced single-strand breaks (SSB) more slowly than the parent line AA8. EM9 was determined to be 1.7-fold more sensitive to killing by 365-nm radiation than AA8 at the 10% survival level, and EM9 had a smaller shoulder region on the survival curve ({alpha} = 1.76) than AA8 ({alpha} = 0.62). No significant differences were found between the cell lines in the initial yields of SSB induced either by {gamma}-radiation (as determined by alkaline sucrose gradient sedimentation) or by 365-nm UVA (as determined by alkaline elution). For measurement of initial SSB, cells were irradiated at 0.5{sup o}C to minimize DNA repair processes. Rejoining of 365-nm induced SSB was measured by irradiating cells at 0.5{sup o}C, allowing them to repair at 37{sup o}C in full culture medium, and then quantitating the remaining SSB by alkaline elution. The repair of these breaks followed biphasic kinetics in both cell lines. EM9 repaired the breaks more slowly (T{sub 1/2} values of 1.3 and 61.3 min) than did AA8 (T{sub 1/2} values of 0.9 and 53.3 min), and EM9 also left more breaks unrepaired 90 min after irradiation (24% vs 8% for AA8). Thus, the sensitivity of EM9 to 365-nm radiation correlated with its deficiency in repairing DNA lesions revealed as SSB in alkaline elution. These results suggest that DNA may be a critical target in 365-nm induced cellular lethality and that the ability of AA8 and EM9 cells to repair DNA strand breaks may be related to their ability to survive 365-nm radiation. (author).

  3. A quantitative AFM analysis of nano-scale surface roughness in various orthodontic brackets.

    Science.gov (United States)

    Lee, Gi-Ja; Park, Ki-Ho; Park, Young-Guk; Park, Hun-Kuk

    2010-10-01

    In orthodontics, the surface roughnesses of orthodontic archwire and brackets affect the effectiveness of arch-guided tooth movement, corrosion behavior, and the aesthetics of orthodontic components. Atomic force microscopy (AFM) measurements were used to provide quantitative information on the surface roughness of the orthodontic material. In this study, the changes in surface roughness of various orthodontic bracket slots before and after sliding movement of archwire in vitro and in vivo were observed through the utilization of AFM. Firstly, we characterized the surface of four types of brackets slots as follows: conventional stainless steel (Succes), conventional ceramic (Perfect), self-ligating stainless steel (Damon) and self-ligating ceramic (Clippy-C) brackets. Succes) and Damon brackets showed relatively smooth surfaces, while Perfect had the roughest surface among the four types of brackets used. Secondly, after in vitro sliding test with beta titanium wire in two conventional brackets (Succes and Perfect), there were significant increases in only stainless steel bracket, Succes. Thirdly, after clinical orthodontic treatment for a maximum of 2 years, the self-ligating stainless steel bracket, Damon, showed a significant increase in surface roughness. But self-ligating ceramic brackets, Clippy-C, represented less significant changes in roughness parameters than self-ligating stainless steel ones. Based on the results of the AFM measurements, it is suggested that the self-ligating ceramic bracket has great possibility to exhibit less friction and better biocompatibility than the other tested brackets. This implies that these bracket slots will aid in the effectiveness of arch-guided tooth movement.

  4. Photovoltaic Wire, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — This Small Business Innovation Research Phase I project will investigate a new architecture for photovoltaic devices based on nanotechnology: photovoltaic wire. The...

  5. Charpak hemispherical wire chamber

    CERN Multimedia

    1970-01-01

    pieces. Mesures are of the largest one. Multi-wire detectors contain layers of positively and negatively charged wires enclosed in a chamber full of gas. A charged particle passing through the chamber knocks negatively charged electrons out of atoms in the gas, leaving behind positive ions. The electrons are pulled towards the positively charged wires. They collide with other atoms on the way, producing an avalanche of electrons and ions. The movement of these electrons and ions induces an electric pulse in the wires which is collected by fast electronics. The size of the pulse is proportional to the energy loss of the original particle.

  6. Adhesives for fixed orthodontic brackets.

    Science.gov (United States)

    Mandall, N A; Millett, D T; Mattick, C R; Hickman, J; Macfarlane, T V; Worthington, H V

    2003-01-01

    Bonding of orthodontic brackets to teeth is important to enable effective and efficient treatment with fixed appliances. The problem is bracket failure during treatment which increases operator chairside time and lengthens treatment time. A prolonged treatment is likely to increase the oral health risks of orthodontic treatment with fixed appliances one of which is irreversible enamel decalcification. To evaluate the effectiveness of different orthodontic adhesives for bonding. Electronic databases: the Cochrane Oral Health Group's Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE and EMBASE. Date of most recent searches: August 2002 (CENTRAL) (The Cochrane Library Issue 2, 2002). Trials were selected if they met the following criteria: randomised controlled trials (RCTs) and controlled clinical trials (CCTs) comparing two different adhesive groups. Participants were patients with fixed orthodontic appliances. The interventions were adhesives that bonded stainless steel brackets to all teeth except the molars. The primary outcome was debond or bracket failure. Data were recorded on decalcification as a secondary outcome, if present. Information regarding methods, participants, interventions, outcome measures and results were extracted in duplicate by pairs of reviewers (Nicky Mandall (NM) and Rye Mattick (CRM); Declan Millett (DTM) and Joy Hickman (JH2)). Since the data were not presented in a form that was amenable to meta-analysis, the results of the review are presented in narrative form only. Three trials satisfied the inclusion criteria. A chemical cured composite was compared with a light cure composite (one trial), a conventional glass ionomer cement (one trial) and a polyacid-modified resin composite (compomer) (one trial). The quality of the trial reports was generally poor. It is difficult to draw any conclusions from this review, however, suggestions are made for methods of improving future research involving

  7. Myofunctional orthodontic for growth child

    Directory of Open Access Journals (Sweden)

    Harun Achmad

    2016-06-01

    Full Text Available Normal growth and development is a concern for every parent! If your child’s teeth are not straight, facial growth and development could also be affected. Current orthodontic treatment can damage teeth, restrict jaw growth and usually is not a stable result. Most importantly, braces and extractions do not address the real causes of crooked teeth and quite often once they are removed, the teeth crowd up again. Studies have shown that teeth will have a tendency to change their positions after treatment hence the potential need for lifetime retainers with traditional orthodontics. Our orthodontic treatment uses myofunctional techniques to address the poor oral habits (known as myofunctional habits that are the real, underlying causes of crooked teeth and uses light, intermittent forces to align the teeth. Myofunctional orthodontic techniques have been practiced by Orthodontists and Dentists around the world for over 25 years. This is done through the use of a series of removable dental appliances that are worn for just 1-2 hours each day and overnight while sleeping. In traditional adult orthodontic treatment, certain factors can contribute to less stable results and likely long term or permanent retainers being required if poor oral habits have not been corrected. Orofacial Myofunctional therapy is a profound process used to address the improper function of the tongue and facial muscles used at rest. Treatment may help address the symptoms of a a wide variety of health issues, ranging from opening airways, to treating headaches, Temporomandibular Joint Disorders (TMD, and poor digestion just to name a few. Myofunctional therapy is the “neuromuscular re-education or re-patterning of the oral and facial muscles”. The therapy includes facial and tongue exercises and techniques that modify the poor habits that affect proper tongue position, improved breathing, chewing, and swallowing.

  8. 1998 wire development workshop proceedings

    International Nuclear Information System (INIS)

    1998-04-01

    This report consists of vugraphs of the presentations at the conference. The conference was divided into the following sessions: (1) First Generation Wire Development: Status and Issues; (2) First Generation Wire in Pre-Commercial Prototypes; (3) Second Generation Wire Development: Private Sector Progress and Issues; (4) Second Generation Wire Development: Federal Laboratories; and (5) Fundamental Research Issues for HTS Wire Development

  9. 1998 wire development workshop proceedings

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1998-04-01

    This report consists of vugraphs of the presentations at the conference. The conference was divided into the following sessions: (1) First Generation Wire Development: Status and Issues; (2) First Generation Wire in Pre-Commercial Prototypes; (3) Second Generation Wire Development: Private Sector Progress and Issues; (4) Second Generation Wire Development: Federal Laboratories; and (5) Fundamental Research Issues for HTS Wire Development.

  10. Orthodontics Align Crooked Teeth and Boost Self-Esteem

    Science.gov (United States)

    ... desktop! more... Orthodontics Align Crooked Teeth and Boost Self- esteem Article Chapters Orthodontics Align Crooked Teeth and Boost Self- esteem print full article print this chapter email this ...

  11. Wire Array Photovoltaics

    Science.gov (United States)

    Turner-Evans, Dan

    Over the past five years, the cost of solar panels has dropped drastically and, in concert, the number of installed modules has risen exponentially. However, solar electricity is still more than twice as expensive as electricity from a natural gas plant. Fortunately, wire array solar cells have emerged as a promising technology for further lowering the cost of solar. Si wire array solar cells are formed with a unique, low cost growth method and use 100 times less material than conventional Si cells. The wires can be embedded in a transparent, flexible polymer to create a free-standing array that can be rolled up for easy installation in a variety of form factors. Furthermore, by incorporating multijunctions into the wire morphology, higher efficiencies can be achieved while taking advantage of the unique defect relaxation pathways afforded by the 3D wire geometry. The work in this thesis shepherded Si wires from undoped arrays to flexible, functional large area devices and laid the groundwork for multijunction wire array cells. Fabrication techniques were developed to turn intrinsic Si wires into full p-n junctions and the wires were passivated with a-Si:H and a-SiNx:H. Single wire devices yielded open circuit voltages of 600 mV and efficiencies of 9%. The arrays were then embedded in a polymer and contacted with a transparent, flexible, Ni nanoparticle and Ag nanowire top contact. The contact connected >99% of the wires in parallel and yielded flexible, substrate free solar cells featuring hundreds of thousands of wires. Building on the success of the Si wire arrays, GaP was epitaxially grown on the material to create heterostructures for photoelectrochemistry. These cells were limited by low absorption in the GaP due to its indirect bandgap, and poor current collection due to a diffusion length of only 80 nm. However, GaAsP on SiGe offers a superior combination of materials, and wire architectures based on these semiconductors were investigated for multijunction

  12. Emprego racional da Biomecânica em Ortodontia: "arcos inteligentes" Biomechanics in Orthodontics: "smart archwires"

    Directory of Open Access Journals (Sweden)

    Marcio Rodrigues de Almeida

    2006-02-01

    Full Text Available A versatilidade biomecânica propiciada pelos "arcos inteligentes" ("smart archwires" permite estratégias para cada tipo de má oclusão, diferenciada por nuances terapêuticas, que por sua vez geram um melhor controle do caso clínico. Os arcos multifuncionais (Arco de intrusão de nitinol CIA® e arco de retração Mushroom Loop® foram idealizados no intuito de simplificar o atendimento clínico do ortodontista que, municiado de recursos diagnósticos apropriados e sólidos, os utiliza como uma ótima alternativa entre as disponíveis para a resolução dos problemas e expectativas dos pacientes. Com o presente artigo procurou-se demonstrar que os arcos de intrusão de nitinol (CIA® e o arco de retração Mushroom Loop® são ferramentas poderosas no tratamento de pacientes com extrações dentárias que requerem um controle de ancoragem mais apropriado, bem como uma retração mais eficaz.The versatility of Biomechanics with the use of "smart wires" allows different strategies in order to achieve a better control in orthodontic cases. Multifunctional orthodontic wires (nickel-titanium intrusion arch-CIA® and M-loop® archwires were developed in order to provide advantages in clinical management for the orthodontists. Since the force system, along with the side effects, of theses wires is well understood it becomes easier to use noncompliance mechanics. The present paper showed the clinical applications of the "smart wires" (nickel-titanium intrusion arch and M-loop archwires capable of simultaneously performing different orthodontic movements in extraction cases which minimize the need for high anchorage control.

  13. The Rev1 interacting region (RIR) motif in the scaffold protein XRCC1 mediates a low-affinity interaction with polynucleotide kinase/phosphatase (PNKP) during DNA single-strand break repair.

    Science.gov (United States)

    Breslin, Claire; Mani, Rajam S; Fanta, Mesfin; Hoch, Nicolas; Weinfeld, Michael; Caldecott, Keith W

    2017-09-29

    The scaffold protein X-ray repair cross-complementing 1 (XRCC1) interacts with multiple enzymes involved in DNA base excision repair and single-strand break repair (SSBR) and is important for genetic integrity and normal neurological function. One of the most important interactions of XRCC1 is that with polynucleotide kinase/phosphatase (PNKP), a dual-function DNA kinase/phosphatase that processes damaged DNA termini and that, if mutated, results in ataxia with oculomotor apraxia 4 (AOA4) and microcephaly with early-onset seizures and developmental delay (MCSZ). XRCC1 and PNKP interact via a high-affinity phosphorylation-dependent interaction site in XRCC1 and a forkhead-associated domain in PNKP. Here, we identified using biochemical and biophysical approaches a second PNKP interaction site in XRCC1 that binds PNKP with lower affinity and independently of XRCC1 phosphorylation. However, this interaction nevertheless stimulated PNKP activity and promoted SSBR and cell survival. The low-affinity interaction site required the highly conserved Rev1-interacting region (RIR) motif in XRCC1 and included three critical and evolutionarily invariant phenylalanine residues. We propose a bipartite interaction model in which the previously identified high-affinity interaction acts as a molecular tether, holding XRCC1 and PNKP together and thereby promoting the low-affinity interaction identified here, which then stimulates PNKP directly. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. Seamless Genetic Conversion of SMN2 to SMN1 via CRISPR/Cpf1 and Single-Stranded Oligodeoxynucleotides in Spinal Muscular Atrophy Patient-Specific Induced Pluripotent Stem Cells.

    Science.gov (United States)

    Zhou, Miaojin; Hu, Zhiqing; Qiu, Liyan; Zhou, Tao; Feng, Mai; Hu, Qian; Zeng, Baitao; Li, Zhuo; Sun, Qianru; Wu, Yong; Liu, Xionghao; Wu, Lingqian; Liang, Desheng

    2018-05-09

    Spinal muscular atrophy (SMA) is a kind of neuromuscular disease characterized by progressive motor neuron loss in the spinal cord. It is caused by mutations in the survival motor neuron 1 (SMN1) gene. SMN1 has a paralogous gene, survival motor neuron 2 (SMN2), in humans that is present in almost all SMA patients. The generation and genetic correction of SMA patient-specific induced pluripotent stem cells (iPSCs) is a viable, autologous therapeutic strategy for the disease. Here, c-Myc-free and non-integrating iPSCs were generated from the urine cells of an SMA patient using an episomal iPSC reprogramming vector, and a unique crRNA was designed that does not have similar sequences (≤3 mismatches) anywhere in the human reference genome. In s