Distinct Roles of FANCO/RAD51C Protein in DNA Damage Signaling and Repair Implications for Fanconi Anemia and Breast Cancer Susceptibility

RAD51C, a RAD51 paralog, has been implicated in homologous recombination (HR), and germ line mutations in RAD51C are known to cause Fanconi anemia (FA)-like disorder and breast and ovarian cancers. The role of RAD51C in the FA pathway of DNA interstrand cross-link (ICL) repair and as a tumor suppressor is obscure. Here, we report that RAD51C deficiency leads to ICL sensitivity, chromatid-type errors, and G(2)/M accumulation, which are hallmarks of the FA phenotype. We find that RAD51C is dispensable for ICL unhooking and FANCD2 monoubiquitination but is essential for HR, confirming the downstream role of RAD51C in ICL repair. Furthermore, we demonstrate that RAD51C plays a vital role in the HR-mediated repair of DNA lesions associated with replication. Finally, we show that RAD51C participates in ICL and double strand break-induced DNA damage signaling and controls intra-S-phase checkpoint through CHK2 activation. Our analyses with pathological mutants of RAD51C that were identified in FA and breast and ovarian cancers reveal that RAD51C regulates HR and DNA damage signaling distinctly. Together, these results unravel the critical role of RAD51C in the FA pathway of ICL repair and as a tumor suppressor.

IN-VITRO CYTOTOXICITY AND CELL CYCLE ANALYSIS OF TWO NOVEL BIS-1, 2, 4-TRIAZOLE DERIVATIVES: 1,4-BIS[5-(5-MERCAPTO-1,3,4-OXADIAZOL-2-yl-METHYL)-THIO-4-(p-TOLYL)-1, 2,4-TRIAZOL-3-yl]-BUTANE (MNP-14) AND 1,4-BIS[5-(CARBETHOXY-METHYL)-THIO-4-(p-ETHOXY PHENYL)-1,2,4-TRIAZOL-3-yl]-BUTANE (MNP-16)

In the present study, we have tested the cytotoxic and DNA damage activity of two novel bis-1,2,4 triazole derivatives, namely 1,4-bis[5-(5-mercapto-1,3,4-oxadiazol-2-yl-methyl)-thio4-(p-tolyl)-1,2 ,4-triazol-3-yl]-butane (MNP-14) and 1,4-bis[5-(carbethoxy-methyl)-thio-4-(p-ethoxy phenyl) -1,2,4-triazol-3-yl]-butane (MNP-16). The effect of these molecules on cellular apoptosis was also determined. The in-vitro cytotoxicity was evaluated by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay as well as Trypan blue dye exclusion methods against human acute lymphoblastic leukemia (MOLT4) and lung cancer cells (A549). Our results showed that MNP-16 induced significant cytotoxicity (IC50 of 3-5 mu M) compared with MNP-14. The cytotoxicity induced by MNP-16 was time and concentration dependent. The cell cycle analysis by flow cytometry (fluorescence-activated cell sorting [FACS]) revealed that though there was a significant increase in the apoptotic population (sub-G1 phase) with an increased concentration of MNP-14 and 16, there was no cell cycle arrest. Further, the comet assay results indicated considerable DNA

Micromechanisms of damage nucleation during contact deformation of columnar multilayer nitride coatings

In order to resolve some missing micromechanistic details regarding contact deformation in nitride multilayer coatings we report here observations from cross-sectional transmission electron microscopy and focused ion beam studies of the Vickers indentations on TiN/TiAlN multilayer films of various total thicknesses as well as bilayer periods. The study of damage induced by contact deformation in a nitride multilayer coating is complemented by stress calculated using an analytical model. Kinked boundaries of sliding columns give rise to cracks which propagate at an angle to the indentation axis under a combination of compressive and shear stresses. It is seen that multilayers provide more distributed columnar sliding, thereby reducing the stress intensity factor for shear cracking, while interfacial dislocations provide a stress relief mechanism by enabling lateral movement of material. (C) 2012 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Ultrasonic Guided Wave Characterization and Damage Detection in Foam-core Sandwich Panel using PWAS and LDV

Lamb wave type guided wave propagation in foam core sandwich structures and detectability of damages using spectral analysis method are reported in this paper. An experimental study supported by theoretical evaluation of the guided wave characteristics is presented here that shows the applicability of Lamb wave type guided ultrasonic wave for detection of damage in foam core sandwich structures. Sandwich beam specimens were fabricated with 10 mm thick foam core and 0.3 mm thick aluminum face sheets. Thin piezoelectric patch actuators and sensors are used to excite and sense guided wave. Group velocity dispersion curves and frequency response of sensed signal are obtained experimentally. The nature of damping present in the sandwich panel is monitored by measuring the sensor signal amplitude at various different distances measured from the center of the linear phased array. Delaminations of increasing width are created and detected experimentally by pitch-catch interrogation with guided waves and wavelet transform of the sensed signal. Signal amplitudes are analyzed for various different sizes of damages to differentiate the damage size/severity. A sandwich panel is also fabricated with a planer dimension of 600 mm x 400 mm. Release film delamination is introduced during fabrication. Non-contact Laser Doppler Vibrometer (LDV) is used to scan the panel while exciting with a surface bonded piezoelectric actuator. Presence of damage is confirmed by the reflected wave fringe pattern obtained from the LDV scan. With this approach it is possible to locate and monitor the damages by tracking the wave packets scattered from the damages.

Guided Wave based Damage Detection in a Composite T-joint using 3D Scanning Laser Doppler Vibrometer

Composite T-joints are commonly used in modern composite airframe, pressure vessels and piping structures, mainly to increase the bending strength of the joint and prevents buckling of plates and shells, and in multi-cell thin-walled structures. Here we report a detailed study on the propagation of guided ultrasonic wave modes in a composite T-joint and their interactions with delamination in the co-cured co-bonded flange. A well designed guiding path is employed wherein the waves undergo a two step mode conversion process, one is due to the web and joint filler on the back face of the flange and the other is due to the delamination edges close to underneath the accessible surface of the flange. A 3D Laser Doppler Vibrometer is used to obtain the three components of surface displacements/velocities of the accessible face of the flange of the T-joint. The waves are launched by a piezo ceramic wafer bonded on to the back surface of the flange. What is novel in the proposed method is that the location of any change in material/geometric properties can be traced by computing a frequency domain power flow along a scan line. The scan line can be chosen over a grid either during scan or during post-processing of the scan data off-line. The proposed technique eliminates the necessity of baseline data and disassembly of structure for structural interrogation.

Evaluation of damage in reinforced concrete bridge beams using acoustic emission technique

Acoustic emission (AE) testing is a well-known method for damage identification of various concrete structures including bridges. This article presents a method to assess damage in reinforced concrete (RC) bridge beams subjected to incremental cyclic loading. The specifications in the standard NDIS-2421 were used to classify the damage in RC bridge beams. Earlier researchers classified the damage occurring in bridge beams by using crack mouth opening displacement (CMOD) and AE released and proposed a standard (NDIS-2421: the Japanese Society for NonDestructive Inspection). In general, multiple cracks take place in RC beams under bending; therefore, utilisation of CMOD for crack detection may not be appropriate. In the present study, the damage in RC beams is classified by using the AE released, deflection, strains in steel and concrete, because the measurement of the strains in steel and concrete is easy and the codes of practice are specified for different limit states (IS-456:2000). The observations made in the present experimental study have some important practical applications in assessing the state of damage of concrete structural members.

Damage-tolerant design optimization of laminated composite structures using dispersion of ply angles by genetic algorithm

This article aims to obtain damage-tolerant designs with minimum weight for a laminated composite structure using genetic algorithm. Damage tolerance due to impacts in a laminated composite structure is enhanced by dispersing the plies such that too many adjacent plies do not have the same angle. Weight of the structure is minimized and the Tsai-Wu failure criterion is considered for the safe design. Design variables considered are the number of plies and ply orientation. The influence of dispersed ply angles on the weight of the structure for a given loading conditions is studied by varying the angles in the range of 0 degrees-45 degrees, 0 degrees-60 degrees and 0 degrees-90 degrees at intervals of 5 degrees and by using specific ply angles tailored to loading conditions. A comparison study is carried out between the conventional stacking sequence and the stacking sequence with dispersed ply angles for damage-tolerant weight minimization and some useful designs are obtained. Unconventional stacking sequence is more damage tolerant than the conventional stacking sequence is demonstrated by performing a finite element analysis under both tensile as well as compressive loading conditions. Moreover, a new mathematical function called the dispersion function is proposed to measure the dispersion of ply angles in a laminate. The approach for dispersing ply angles to achieve damage tolerance is especially suited for composite material design space which has multiple local minima.

Guided wave based detection of damage in honeycomb core sandwich structures

We report on the Lamb wave type guided wave propagation in honeycomb core sandwich structures. An experimental study supported by theoretical evaluation of the guided wave characteristics is presented that proves the potential of Lamb wave type guided wave for detection of damage in sandwich structures. A sandwich panel is fabricated with planar dimension of 600 mm x 600 mm, having a core thickness of 7 mm, cell size of 5 mm and 0.1 mm thick aluminum face sheets. Thin piezoelectric patch actuators and sensors are used to excite and sense a frequency band limited guided wave with a central frequency. A linear phased array of piezoelectric patch actuators is used to achieve higher signal strength and directivity. Group velocity dispersion curves and corresponding frequency response of sensed signal are obtained experimentally. Linearity between the excitation signal amplitude and the corresponding sensed signal amplitude is found for certain range of parameters. The nature of damping present in the sandwich panel is monitored by measuring the sensor signal amplitude at various different distances measured from the center of the linear phased array. Indentation and low velocity impact induced damages of increasing diameter covering several honeycomb cells are created. Crushing of honeycomb core with rupture of face sheet is observed while introducing the damage. The damages are then detected experimentally by pitch-catch interrogation with guided waves and wavelet transform of the sensed signal. Signal amplitudes are analyzed for various different sizes of damages to differentiate the damage size/severity. Monotonic changes in the sensor signal amplitude due to increase in the damage size has been established successfully. With this approach it is possible to locate and monitor the damages with the help of phased array and by tracking the wave packets scattered from the damages. (C) 2012 Elsevier Ltd. All rights reserved.

Damage limit states of reinforced concrete beams subjected to incremental cyclic loading using relaxation ratio analysis of AE parameters

This paper presents an experimental study on damage assessment of reinforced concrete (RC) beams subjected to incremental cyclic loading. During testing acoustic emissions (AEs) were recorded. The analysis of the AE released was carried out by using parameters relaxation ratio, load ratio and calm ratio. Digital image correlation (DIC) technique and tracking with available MATLAB program were used to measure the displacement and surface strains in concrete. Earlier researchers classified the damage in RC beams using Kaiser effect, crack mouth opening displacement and proposed a standard. In general (or in practical situations), multiple cracks occur in reinforced concrete beams. In the present study damage assessment in RC beams was studied according to different limit states specified by the code of practice IS-456:2000 and AE technique. Based on the two ratios namely load ratio and calm ratio and when the deflection reached approximately 85% of the maximum allowable deflection it was observed that the RC beams were heavily damaged. The combination of AE and DIC techniques has the potential to provide the state of damage in RC structures.

Fatigue damage assessment in cracked RC beams

In this work, an attempt has been made to assess the fatigue life of reinforced concrete beams, by proposing a crack propagation law which accounts for parameters such as fracture toughness, crack length, loading ratio and structural size. A numerical procedure is developed to compute fatigue life of RC beams. The predicted results are compared with the available experimental data in the literature and seen to agree reasonably well. Further, in order to assess the remaining life of an RC member, the moment carrying capacity is determined as a function of crack extension, based on the crack tip opening displacement and residual strength of the member is computed at an event of unstable fracture.

Damage identification of beam-like structures with contiguous and distributed damage

Structural health monitoring of existing infrastructure is currently an active field of research, where elaborate experimental programs and advanced analytical methods are used in identifying the current state of health of critical structures. Change of static deflection as the indicator of damage is the simplest tool in a structural health monitoring scenario of bridges that is least exploited in damage identification strategies. In this paper, some simple and elegant equations based on loss of symmetry due to damage are derived and presented for identification of damage in a bridge girder modeled as a simply supported beam using changes in static deflections and dynamic parameters. A single contiguous and distributed damage, typical of reinforced or prestressed concrete structures, is assumed for the structure. The methodology is extended for a base-line-free as well as base-line-inclusive measurement. Measurement strategy involves application of loads only at two symmetric points one at a time and deflection measurements at those symmetric points as well as at the midspan of the beam. A laboratory-based experiment is used to validate the approach. Copyright (c) 2012 John Wiley & Sons, Ltd.

Combination of single walled carbon nanotubes/graphene oxide with paclitaxel: a reactive oxygen species mediated synergism for treatment of lung cancer

Heterogeneity in tumors has led to the development of combination therapies that enable enhanced cell death. Previously explored combination therapies mostly involved the use of bioactive molecules. In this work, we explored a non-conventional strategy of using carbon nanostructures (CNs) single walled carbon nanotube (SWNT) and graphene oxide (GO)] for potentiating the efficacy of a bioactive molecule paclitaxel (Tx)] for the treatment of lung cancer. The results demonstrated enhanced cell death following combination treatment of SWNT/GO and Tx indicating a synergistic effect. In addition, synergism was abrogated in the presence of an anti-oxidant, N-acetyl cysteine (NAC), and was therefore shown to be reactive oxygen species (ROS) dependent. It was further demonstrated using bromodeoxyuridine (BrdU) incorporation assay that treatment with CNs was associated with enhanced mitogen associated protein kinase (MAPK) activation that was ROS mediated. Hence, these results for the first time demonstrated the potential of SWNT/GO as co-therapeutic agents with Tx for the treatment of lung cancer.

Influence of crack parameters on wavelet energy correlated damage index

This paper illustrates a Wavelet Coefficient based approach using experiments to understand the sensitivity of ultrasonic signals due to parametric variation of a crack configuration in a metal plate. A PZT patch sensor/actuator system integrated to a metal plate with through-thickness crack is used. The proposed approach uses piezoelectric patches, which can be used to both actuate and sense the ultrasonic signals. While this approach leads to more flexibility and reduced cost for larger scalability of the sensor/actuator network, the complexity of the signals increases as compared to what is encountered in conventional ultrasonic NDE problems using selective wave modes. A Damage Index (DI) has been introduced, which is function of wavelet coefficient. Experiments have been carried out for various crack sizes, crack orientations and band-limited tone-burst signal through FIR filter. For a 1 cm long crack interrogated with 20 kHz tone-burst signal, the Damage Index (DI) for the horizontal crack orientation increases by about 70% with respect to that for 135 degrees oriented crack and it increases by about 33% with respect to the vertically oriented crack. The detailed results reported in this paper is a step forward to developing computational schemes for parametric identification of damage using sensor/actuator network and ultrasonic wave.

Guided Ultrasonic Wave Propagation through Inaccessible Damage in a Folded Plate using Sensor-Actuator Network

Rapid diagnostics and virtual imaging of damages in complex structures like folded plate can help reduce the inspection time for guided wave based NDE and integrated SHM. Folded plate or box structure is one of the major structural components for increasing the structural strength. Damage in the folded plate, mostly in the form of surface breaking cracks in the inaccessible zone is a usual problem in aerospace structures. One side of the folded plate is attached (either riveted or bonded) to adjacent structure which is not accessible for immediate inspection. The sensor-actuator network in the form of a circular array is placed on the accessible side of the folded plate. In the present work, a circular array is employed for scanning the entire folded plate type structure for damage diagnosis and wave field visualization of entire structural panel. The method employs guided wave with relatively low frequency bandwidth of 100-300 kHz. Change in the response signal with respect to a baseline signal is used to construct a quantitative relationship with damage size parameters. Detecting damage in the folded plate by using this technique has significant potential for off-line and on-line SHM technologies. By employing this technique, surface breaking cracks on inaccessible face of the folded plate are detected without disassembly of structure in a realistic environment.

A Guided Wave approach for the detection of damage in a structure having elements with periodic damage

A wave propagation based approach for the detection of damage in components of structures having periodic damage has been proposed. Periodic damage pattern may arise in a structure due to periodicity in geometry and in loading. The method exploits the Block-Floquet band formation mechanism, a feature specific to structures with periodicity, to identify propagation bands (pass bands) and attenuation bands (stop bands) at different frequency ranges. The presence of damage modifies the wave propagation behaviour forming these bands. With proper positioning of sensors a damage force indicator (DFI) method can be used to locate the defect at an accuracy level of sensor to sensor distance. A wide range of transducer frequency may be used to obtain further information about the shape and size of the damage. The methodology is demonstrated using a few 1-D structures with different kinds of periodicity and damage. For this purpose, dynamic stiffness matrix is formed for the periodic elements to obtain the dispersion relationship using frequency domain spectral element and spectral super element method. The sensitivity of the damage force indicator for different types of periodic damages is also analysed.