Segregational Stability Research Articles

Effects of three different nucleoid-associated proteins encoded on IncP-7 plasmid pCAR1 on host Pseudomonas putida KT2440.

Nucleoid-associated proteins (NAPs), which fold bacterial DNA and influence gene transcription, are considered to be global transcriptional regulators of genes on both plasmids and the host chromosome. Incompatibility P-7 group plasmid pCAR1 carries genes encoding three NAPs: H-NS family protein Pmr, NdpA-like protein Pnd, and HU-like protein Phu. In this study, the effects of single or double disruption of pmr, pnd, and phu were assessed in host Pseudomonas putida KT2440. When pmr and pnd or pmr and phu were simultaneously disrupted, both the segregational stability and the structural stability of pCAR1 were markedly decreased, suggesting that Pmr, Pnd, and Phu act as plasmid-stabilizing factors in addition to their established roles in replication and partition systems. The transfer frequency of pCAR1 was significantly decreased in these double mutants. The segregational and structural instability of pCAR1 in the double mutants was recovered by complementation of pmr, whereas no recovery of transfer deficiency was observed. Comprehensive phenotype comparisons showed that the host metabolism of carbon compounds, which was reduced by pCAR1 carriage, was restored by disruption of the NAP gene(s). Transcriptome analyses of mutants indicated that transcription of genes for energy production, conversion, inorganic ion transport, and metabolism were commonly affected; however, how their products altered the phenotypes of mutants was not clear. The findings of this study indicated that Pmr, Pnd, and Phu act synergistically to affect pCAR1 replication, maintenance, and transfer, as well as to alter the host metabolic phenotype.

Characterization of a minimal pKW2124 replicon from Weissella cibaria KLC140 and its application for the construction of the Weissella expression vector pKUCm1.

A 2.1-kb plasmid was previously isolated from Weissella cibaria KLC140 in kimchi and cloned into pUC19 along with the slpA and gfp genes, resulting in an 8.6-kb pKWCSLGFP construct for use as a novel surface display vector. To reduce the size of the vector, the minimal replicon of pKW2124 was determined. The pKW2124 plasmid contains a putative origin of replication (ori), a potential ribosomal binding site (RBS), and the repA gene encoding a plasmid replication protein. To conduct the minimal replicon experiment, four different PCR products (MR1, ori+RBS+repA; MR2, RBS+repA; MR2’, repA; MR3, fragment of repA) were obtained and cloned into pUC19 (pKUCm1, pKUCm2, pKUCm2’, and pKUCm3, respectively) containing the chloramphenicol acetyltransferase (CAT) gene. These constructed vectors were electroporated into W. confusa ATCC 10881 with different transformation efficiencies of 1.5 × 105 CFU/μg, 1.3 × 101 CFU/μg, and no transformation, respectively, suggesting that the putative ori, RBS, and repA gene are essential for optimum plasmid replication. Subsequent segregational plasmid stability testing of pKUCm1 and pKUCm2 showed that the vector pKUCm1 is highly stable up to 100 generations but pKUCm2 was completely lost after 60 generations, suggesting that the putative ori may be important for plasmid stability in the host strain. In addition, a host range test of pKUCm1 revealed that it has a broad host range spectrum including Weissella, Lactococcus, Leuconostoc, and even Lactobacillus. To verify the application of pKUCm1, the β-galactosidase gene and its promoter region from W. cibaria KSD1 were cloned in the vector, resulting in pKUGal. Expression of the β-galactosidase gene was confirmed using blue-white screening after IPTG induction. The small and stable pKUGal vector will be useful for gene transfer, expression, and manipulation in the Weissella genome and in other lactic acid bacteria.

ASAR15, A cis-acting locus that controls chromosome-wide replication timing and stability of human chromosome 15.

DNA replication initiates at multiple sites along each mammalian chromosome at different times during each S phase, following a temporal replication program. We have used a Cre/loxP-based strategy to identify cis-acting elements that control this replication-timing program on individual human chromosomes. In this report, we show that rearrangements at a complex locus at chromosome 15q24.3 result in delayed replication and structural instability of human chromosome 15. Characterization of this locus identified long, RNA transcripts that are retained in the nucleus and form a “cloud” on one homolog of chromosome 15. We also found that this locus displays asynchronous replication that is coordinated with other random monoallelic genes on chromosome 15. We have named this locus ASynchronous replication and Autosomal RNA on chromosome 15, or ASAR15. Previously, we found that disruption of the ASAR6 lincRNA gene results in delayed replication, delayed mitotic condensation and structural instability of human chromosome 6. Previous studies in the mouse found that deletion of the Xist gene, from the X chromosome in adult somatic cells, results in a delayed replication and instability phenotype that is indistinguishable from the phenotype caused by disruption of either ASAR6 or ASAR15. In addition, delayed replication and chromosome instability were detected following structural rearrangement of many different human or mouse chromosomes. These observations suggest that all mammalian chromosomes contain similar cis-acting loci. Thus, under this scenario, all mammalian chromosomes contain four distinct types of essential cis-acting elements: origins, telomeres, centromeres and “inactivation/stability centers”, all functioning to promote proper replication, segregation and structural stability of each chromosome.

Effects of Carbon Pre-Germanidation Implantation on the Thermal Stability of NiGe and Dopant Segregation on Both n- and p-Type Ge Substrate

In this work, the effects of carbon pre-geramanidation implantation on the thermal stability of NiGe and dopant segregation on both n-type and p-type Ge substrate were investigated systematically. As-prepared NiGe films with carbon pre-germanidation implantation to different doses were characterized by means of sheet resistance measurement, X-ray diffraction (XRD), scanning electron microscopy (SEM), cross-sectional transmission electron microscope (X-TEM) and secondary ion mass spectroscopy (SIMS). The presence of carbon is proved to improve the thermal stability of NiGe formed on both n- and p-type Ge significantly, as well as to lead to dopant segregation (DS) of P and B at the NiGe/Ge interface. The homogeneous distribution of C within NiGe films and stuffing of C atoms at the NiGe/Ge interface is responsible for the enhanced thermal stability of NiGe and DS of P and B during germanidation process.

Impact of galactosylceramides on the nanomechanical properties of lipid bilayer models: an AFM-force spectroscopy study.

Galactosylceramides (GalCer) are glycosphingolipids bound to a monosaccharide group, responsible for inducing extensive hydrogen bonds that yield their alignment and accumulation in the outer leaflet of the biological membrane together with cholesterol (Chol) in rafts. In this work, the influence of GalCer on the nanomechanical properties of supported lipid bilayers (SLBs) based on DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) and DLPC (1,2-didodecanoyl-sn-glycero-3-phosphocoline) as model systems was assessed. Phosphatidylcholine (PC):GalCer SLBs were characterized by means of differential scanning calorimetry (DSC) and atomic force microscopy (AFM), in both imaging and force spectroscopy (AFM-FS) modes. Comparing both PC systems, we determined that the behaviour of SLB mixtures is governed by the PC phase-like state at the working temperature. While a phase segregated system is observed for DLPC:GalCer SLBs, GalCer are found to be dissolved in DPPC SLBs for GalCer contents up to 20 mol%. In both systems, the incorporation of GalCer intensifies the nanomechanical properties of SLBs. Interestingly, segregated domains of exceptionally high mechanical stability are formed in DLPC:GalCer SLBs. Finally, the role of 20 mol% Chol in GalCer organization and function in the membranes was assessed. Both PC model systems displayed phase segregation and remarkable nanomechanical stability when GalCer and Chol coexist in SLBs.

Resveratrol production in bioreactor: Assessment of cell physiological states and plasmid segregational stability

Resveratrol is a plant secondary metabolite commonly found in peanuts and grapevines with significant health benefits. Recombinant organisms can produce large amounts of resveratrol and, in this work, Escherichia coli BW27784 was used to produce resveratrol in bioreactors while monitoring cell physiology and plasmid stability through flow cytometry and real-time qPCR, respectively. Initially, the influence of culture conditions and precursor addition was evaluated in screening assays and the data gathered was used to perform the bioreactor assays, allowing the production of 160μg/mL of resveratrol. Cellular physiology and plasmid instability affected the final resveratrol production, with lower viability and plasmid copy numbers associated with lower yields. In sum, this study describes new tools to monitor the bioprocess, evaluating the effect of culture conditions, and its correlation with cell physiology and plasmid segregational stability, in order to define a viable and scalable bioprocess to fulfill the need for larger quantities of resveratrol.

The stability region of the Streptomyces lividans plasmid pIJ101 encodes a DNA-binding protein recognizing a highly conserved short palindromic sequence motif.

Conjugation is a driving force in the evolution and shaping of bacterial genomes. In antibiotic producing streptomycetes even small plasmids replicating via the rolling-circle mechanism are conjugative. Although they encode only genes involved in replication and transfer, the molecular function of most plasmid encoded proteins is unknown. In this work we show that the conjugative plasmid pIJ101 encodes an overlooked protein, SpdA2. We show that SpdA2 is a DNA binding protein which specifically recognizes a palindromic DNA sequence (sps). sps is localized within the spdA2 coding region and highly conserved in many Streptomyces plasmids. Elimination of the palindrome or deletion of spdA2 in plasmid pIJ303 did not interfere with conjugative plasmid transfer or pock formation, but affected segregational stability.

Stability of vacancy-hydrogen clusters in nickel from first-principles calculations

The interactions of hydrogen (H) atoms with vacancies are investigated by means of ab initio calculations. The lowest segregation energies are −0.27 and −0.41eV at single vacancies and divacancies, respectively. These values are in excellent agreement with those corresponding to the two characteristic peaks of the thermal desorption spectra. The microscopic interpretation of the experimental data is therefore clarified. An energetic model is built from the ab initio data and used to study the influence of H bulk concentration and temperature on the concentration of vacancy-H clusters. Analytical expressions, validated by Monte Carlo simulations, are given. The mean vacancy occupation and the H-induced vacancy enrichment are calculated at two temperatures representative of H embrittlement experiments and stress corrosion cracking at high temperatures. The stability domain of VH6 clusters is found to significantly overlap with the experimental conditions for embrittlement. Therefore, vacancy clustering at high concentrations can be qualitatively discussed based on VH6–VH6 interactions that are found weakly repulsive. Consequences on H damage in Ni are discussed. The effect of metal vibrations on segregation and local hydride stability is qualitatively evaluated by off-lattice Monte Carlo simulations using a semi-empirical Ni–H potential. They are shown to shift local hydride stability towards higher H concentrations.

Differential recombination dynamics within the MHC of macaque species.

A panel of 15 carefully selected microsatellites (short tandem repeats, STRs) has allowed us to study segregation and haplotype stability in various macaque species. The STRs span the major histocompatibility complex (MHC) region and map in more detail from the centromeric part of the Mhc-A to the DR region. Two large panels of Indian rhesus and Indonesian/Indochinese cynomolgus macaques have been subjected to pedigree analysis, allowing the definition of 161 and 36 different haplotypes and the physical mapping of 10 and 5 recombination sites, respectively. Although most recombination sites within the studied section of the Indian rhesus monkey MHC are situated between the Mhc-A and Mhc-B regions, the resulting recombination rate for this genomic segment is low and similar to that in humans. In contrast, in Indonesian/Indochinese macaques, two recombination sites, which appear to be absent in rhesus macaques, map between the class III and II regions. As a result, the mean recombination frequency of the core MHC, Mhc-A to class II, is higher in Indonesian/Indochinese cynomolgus than in Indian rhesus macaques, but as such is comparable to that in humans. The present communication demonstrates that the dynamics of recombination ‘hot/cold spots’ in the MHC, as well as their frequencies, may differ substantially between highly related macaque species.Electronic supplementary materialThe online version of this article (doi:10.1007/s00251-014-0783-4) contains supplementary material, which is available to authorized users.

A bacteriocin gene cluster able to enhance plasmid maintenance in Lactococcus lactis.

BackgroundLactococcus lactis is widely used as a dairy starter and has been extensively studied. Based on the acquired knowledge on its physiology and metabolism, new applications have been envisaged and there is an increasing interest of using L. lactis as a cell factory. Plasmids constitute the main toolbox for L. lactis genetic engineering and most rely on antibiotic resistant markers for plasmid selection and maintenance. In this work, we have assessed the ability of the bacteriocin Lactococcin 972 (Lcn972) gene cluster to behave as a food-grade post-segregational killing system to stabilize recombinant plasmids in L. lactis in the absence of antibiotics. Lcn972 is a non-lantibiotic bacteriocin encoded by the 11-kbp plasmid pBL1 with a potent antimicrobial activity against Lactococcus.ResultsAttempts to clone the full lcn972 operon with its own promoter (P972), the structural gene lcn972 and the immunity genes orf2-orf3 in the unstable plasmid pIL252 failed and only plasmids with a mutated promoter were recovered. Alternatively, cloning under other constitutive promoters was approached and achieved, but bacteriocin production levels were lower than those provided by pBL1. Segregational stability studies revealed that the recombinant plasmids that yielded high bacteriocin titers were maintained for at least 200 generations without antibiotic selection. In the case of expression vectors such as pTRL1, the Lcn972 gene cluster also contributed to plasmid maintenance without compromising the production of the fluorescent mCherry protein. Furthermore, unstable Lcn972 recombinant plasmids became integrated into the chromosome through the activity of insertion sequences, supporting the notion that Lcn972 does apply a strong selective pressure against susceptible cells. Despite of it, the Lcn972 gene cluster was not enough to avoid the use of antibiotics to select plasmid-bearing cells right after transformation.ConclusionsInserting the Lcn972 cluster into segregational unstable plasmids prevents their lost by segregation and probable could be applied as an alternative to the use of antibiotics to support safer and more sustainable biotechnological applications of genetically engineered L. lactis.

The 68-kDa Telomeric Repeat Binding Factor 1 (TRF1)-associated Protein (TAP68) Interacts with and Recruits TRF1 to the Spindle Pole during Mitosis

The telomere capping protein TRF1 is a component of the multiprotein complex "shelterin," which organizes the telomere into a high order structure. Besides telomere maintenance, telomere-associated proteins also have nontelomeric functions. For example, tankyrase 1 and TRF1 are required for the maintenance of faithful mitotic progression. However, the functional relevance of their centrosomal localization has not been established. Here, we report the identification of a TRF1-binding protein, TAP68, that interacts with TRF1 in mitotic cells. TAP68 contains two coiled-coil domains and a structural maintenance of chromosome motifs and co-localizes with TRF1 to telomeres during interphase. Immediately after nuclear envelope breakdown, TAP68 translocates toward the spindle poles followed by TRF1. Dissociation of TAP68 from the telomere is concurrent with the Nek2A-dependent phosphorylation at Thr-221. Biochemical characterization demonstrated that the first coiled-coil domain of TAP68 binds and recruits TRF1 to the centrosome. Inhibition of TAP68 expression by siRNA blocked the localization of TRF1 and tankyrase 1 to the centrosome. Furthermore, siRNA-mediated depletion of TAP68 perturbed faithful chromosome segregation and genomic stability. These findings suggest that TAP68 functions in mediating TRF1-tankyrase 1 localization to the centrosome and in mitotic regulation.

Antibiotic-free segregational plasmid stabilization in Escherichia coli owing to the knockout of triosephosphate isomerase (tpiA).

BackgroundSegregational stability of plasmids is of major concern for recombinant bacterial production strains. One of the best strategies to counteract plasmid loss is the use of auxotrophic mutants which are complemented with the lacking gene along with the product-relevant ones. However, these knockout mutants often show unwanted growth in complex standard media or no growth at all under uncomplemented conditions. This led to the choice of a gene for knockout that only connects two essential arms of an essential metabolic pathway – the glycolysis.ResultsTriosephosphate isomerase was chosen because its knockout will have a tremendous effect on growth on glucose as well as on glycerol. On glycerol the effect is almost absolute whereas on glucose growth is still possible, but with considerably lower rate than usual. This feature is essential because it may render cloning easier. This enzymatic activity was successfully tested as an alternative to antibiotic-based plasmid selection. Expression of a model recombinant β-glucanase in continuous cultivation was possible with stable maintenance of the plasmid. In addition, the complementation of tpiA knockout strains by the corresponding plasmids and their growth characteristics were tested on a series of complex and synthetic media. The accumulation of methylglyoxal during the growth of tpiA-deficient strains was shown to be a possible cause for the growth disadvantage of these strains in comparison to the parent strain for the Keio Collection strain or the complemented knock-out strain.ConclusionThrough the use of this new auxotrophic complementation system, antibiotic-free cloning and selection of recombinant plasmid were possible. Continuous cultivation and recombinant protein expression with high segregational stability over an extended time period was also demonstrated.

Figure-Ground Processing during Fixational Saccades in V1: Indication for Higher-Order Stability

In a typical visual scene we continuously perceive a "figure" that is segregated from the surrounding "background" despite ongoing microsaccades and small saccades that are performed when attempting fixation (fixational saccades [FSs]). Previously reported neuronal correlates of figure-ground (FG) segregation in the primary visual cortex (V1) showed enhanced activity in the "figure" along with suppressed activity in the noisy "background." However, it is unknown how this FG modulation in V1 is affected by FSs. To investigate this question, we trained two monkeys to detect a contour embedded in a noisy background while simultaneously imaging V1 using voltage-sensitive dyes. During stimulus presentation, the monkeys typically performed 1-3 FSs, which displaced the contour over the retina. Using eye position and a 2D analytical model to map the stimulus onto V1, we were able to compute FG modulation before and after each FS. On the spatial cortical scale, we found that, after each FS, FG modulation follows the stimulus retinal displacement and "hops" within the V1 retinotopic map, suggesting visual instability. On the temporal scale, FG modulation is initiated in the new retinotopic position before it disappeared from the old retinotopic position. Moreover, the FG modulation developed faster after an FS, compared with after stimulus onset, which may contribute to visual stability of FG segregation, along the timeline of stimulus presentation. Therefore, despite spatial discontinuity of FG modulation in V1, the higher-order stability of FG modulation along time may enable our stable and continuous perception.

Ethnic residential segregation stability in England, 1991–2001

The residential arrangements of ethnic groups became the subject of political interest when they were identified as a feature of urban areas that experienced unrest in 2001. Residential segregation was framed as both problematic for community relations and a cause of economic inequalities (Cantle, 2001; Denham, 2001). This article presents evidence that ethnic residential segregation in England was not increasing between 1991 and 2001 and that there was a trend for local authority areas to become more similar at a relatively low level of segregation. At neighbourhood level there is little evidence to regard ethnic residential segregation as a problem despite the policy priority it came to have.

Abstract A28: Analyses of FOXM1 isoforms in ovarian cancer

Abstract Introduction & Objectives: Epithelial ovarian cancer (OC) is the most lethal gynaecological malignancy and the fifth leading cause of cancer death in women in the Western world. Forkhead box M1 (FOXM1) transcription factor was already been described to be overexpressed in various malignancies, playing a key role in cell-cycle progression, chromosomal segregation and genomic stability, and oxidative stress. Moreover, it can also promote drug resistance to herceptin, paclitaxel and cisplatin in breast cancer cells. A recent report from our group has shown, for the first time, that FOXM1 is also overexpressed in OC, being associated with tumor grade. This result seem to indicate an interesting role of this molecule in the progression and resistance to therapy in OC. Our aim is to characterize FOXM1 isoforms in OC through the use of human tissue samples. Material & Methods: Fresh ovarian primary tumors, ascites and peritoneal metastases from patients suffering advanced serous OC were collected from surgery room and processed for microarray analysis to detect FOXM1 expression levels. FOXM1 isoforms were analysed at mRNA expression level by using RTqPCR in a set of new frozen tissue samples. Results: We found that FOXM1 overexpression is increased in OC peritoneal metastases. Now, we are characterizing the isoforms of FOXM1 in primary, ascites and metastatic tumors. Nevertheless, more samples need to be analysed in order to have conclusive results. Conclusions: The present study highlights the role of FOXM1 isoforms in OC dissemination and pursues the idea to elucidate the biologic role of FOXM1 in OC tumorigenesis. Citation Format: Tatiana Altadill, Marina Rigau, Blanca Majem, Lucia Lanau, Josep Castellví, Ángel García, Assumpció Pérez-Benavente, José Luis Sánchez, Silvia Cabrera, Jordi Xercavins, Josep Maria del Campo, Antonio Gil-Moreno, Marta Llauradó, Jaume Reventós. Analyses of FOXM1 isoforms in ovarian cancer. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr A28.

Toxin–antitoxin-stabilized reporter plasmids for biophotonic imaging of Group A streptococcus

Bioluminescence is a rapid and cost-efficient optical imaging technology that allows the detection of bacteria in real-time during disease development. Here, we report a novel strategy to generate a wide range of bioluminescent group A streptococcus (GAS) strains by using a toxin-antitoxin-stabilized plasmid. The bacterial luciferin-luciferase operon (lux) or the firefly luciferase gene (ffluc) was introduced into GAS via a stabilized plasmid. The FFluc reporter gave significantly stronger bioluminescent signals than the Lux reporter, and was generally more stable. Plasmid-based luciferase reporters could easily be introduced into a variety of GAS strains and the signals correlated linearly with viable cell counts. Co-expression of the streptococcal ω-ε-ζ toxin-antitoxin operon provided segregational stability in the absence of antibiotics for at least 17 passages in vitro and up to 7days in a mouse infection model. In addition, genome-integrated reporter constructs were also generated by site-specific recombination, but were found to be technically more challenging. The quick and efficient generation of various M-type GAS strains expressing plasmid-based luciferase reporters with comparable and quantifiable bioluminescence signals allows for comparative analysis of different GAS strains in vitro and in vivo.

Estimation of grain boundary segregation enthalpy and its role in stable nanocrystalline alloy design

Abstract

Abstract B16: Cells containing mutations in the cohesin component, STAG2, are sensitive to PARP inhibition

Abstract Synthetic lethality (SL) has been proposed as a strategy to specifically target cells harboring cancer mutations while leaving unmutated, non-cancer cells unaffected, thus minimizing drug side effects. One genetic vulnerability common to many types of cancer is chromosome instability (or CIN). CIN can lead to increased genetic mutation and aneuploidy in cancer cells thereby distinguishing them from their normal neighbors and making it a possible candidate phenotype to target by SL. Here, we focus on the multifunctional cohesin complex that is critical for accurate chromosome segregation and genome stability because it tethers sister chromatids together after DNA replication and maintains cohesion until their separation in anaphase. Although several subunits of cohesin have been shown to be mutated in tumors, recent data identifies a cohesin component, STAG2 (Stromal Antigen 2 or SA-2) as a highly mutated gene in several types of cancer including Ewing's sarcoma, melanoma and glioblastomas (GBMs). As a previous study in our lab had shown that knockdown of any one of the core cohesion subunits SMC1A, SMC3 or RAD21 can render cells sensitive to the poly ADP-ribose polymerase (PARP) inhibitor olaparib, we sought to determine if cell lines harboring cancer-specific STAG2 mutations showed similar sensitivity. Using paired GBM cell lines containing either a truncated STAG2 or wild-type STAG2 knock-in, we found that STAG2 mutation status was, indeed, associated with significantly decreased proliferation in the presence of PARP inhibition. Interestingly, olaparib treatment led to an accumulation of cells in the G2 phase of the cell cycle and an increase in DNA damage markers. Furthermore, in the presence of increasing concentrations of olaparib, a higher percentage of STAG2-mutated cells had micronuclei and fragmented nuclei compared to their wild-type STAG2 knock-in counterparts. Taken together, these data suggest that the DNA repair enzyme PARP may be a potential therapeutic SL target for tumors harboring mutations in STAG2. Citation Format: Melanie L. Bailey, Nigel J. O'Neil, Derek M. van Pel, Phil Hieter. Cells containing mutations in the cohesin component, STAG2, are sensitive to PARP inhibition. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Synthetic Lethal Approaches to Cancer Vulnerabilities; May 17-20, 2013; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(5 Suppl):Abstract nr B16.

Materials challenges in nuclear energy

Nuclear power currently provides about 13% of electrical power worldwide, and has emerged as a reliable baseload source of electricity. A number of materials challenges must be successfully resolved for nuclear energy to continue to make further improvements in reliability, safety and economics. The operating environment for materials in current and proposed future nuclear energy systems is summarized, along with a description of materials used for the main operating components. Materials challenges associated with power uprates and extensions of the operating lifetimes of reactors are described. The three major materials challenges for the current and next generation of water-cooled fission reactors are centered on two structural materials aging degradation issues (corrosion and stress corrosion cracking of structural materials and neutron-induced embrittlement of reactor pressure vessels), along with improved fuel system reliability and accident tolerance issues. The major corrosion and stress corrosion cracking degradation mechanisms for light-water reactors are reviewed. The materials degradation issues for the Zr alloy-clad UO2 fuel system currently utilized in the majority of commercial nuclear power plants are discussed for normal and off-normal operating conditions. Looking to proposed future (Generation IV) fission and fusion energy systems, there are five key bulk radiation degradation effects (low temperature radiation hardening and embrittlement; radiation-induced and -modified solute segregation and phase stability; irradiation creep; void swelling; and high-temperature helium embrittlement) and a multitude of corrosion and stress corrosion cracking effects (including irradiation-assisted phenomena) that can have a major impact on the performance of structural materials.

Effect of mineral admixtures on fluidity and stability of self-consolidating mortar subjected to prolonged mixing time

During continuous mixing of concrete in a truck mixer for its hauling to a construction site, concrete should remain workable without any signs of instability. Good workability at construction site is essential for high quality concrete since concretes of bad workability are prone to yield low strength and poor durability properties. This paper presents the effects of water to binder ratio, binary and ternary blends use of fly ash (FA) and metakaolin (MK) on fluidity, viscosity, and stability of self-consolidating mortars (SCMs) subjected to prolonged mixing time. The obtained results indicate that, by prolonged mixing time, flocculated cement particles are dispersed and as a result, fluidity is increased and so, the risk of instability increases. In addition, increasing FA content from 0 in the reference mixture to 20% and 50% of total binder mass increased the segregation index by about 48% and 160%, respectively. Furthermore, mixtures containing MK and FA have higher fluid capacity without any signs of instability. The addition of 7.5% MK has significantly improved the overall performance of the mixture, including the segregation stability.