Characterisation of interfacial segregation to Cu-enriched precipitates in two thermally aged reactor pressure vessel steel welds.

To understand the contribution of long term thermal ageing to Reactor Pressure Vessel (RPV) embrittlement two high Cu steel welds with different Ni contents were thermally aged for times up to 100,000 h at 330 °C and 365 °C. Microstructural characterisation using Atom Probe Tomography was performed. Thermal ageing produced a high number density of nano-scale Cu-enriched precipitates. The precipitate-matrix interfaces were enriched in Ni, Mn and Si. The characterisation of these interfaces using a double cluster search approach is the subject of this work. The interface region around thermally-induced precipitates was found to be wider in steels with higher bulk Ni contents and where precipitates had larger core radii. The effect of ageing temperature on interface width was small when comparing precipitates of equal core radius. The narrower interface width in the lower Ni steels is reflected in the composition of the interface, which has a lower Ni content than in the higher Ni material. The reduction in interfacial energy due to the segregation of Ni, Mn and Si has been calculated and shows enhanced reductions in interfacial energy with increasing precipitate size, but no obvious effect of temperature.

A comparison of the structure of solute clusters formed during thermal ageing and irradiation.

Nanometre scale clusters form in Cu-containing reactor pressure vessel (RPV) steels during neutron irradiation. These clusters have a deleterious effect on mechanical properties, which can result in embrittlement and limit the reactor operating life. Thermal ageing of RPV steels can also induce the formation of solute clusters but it is not clear how similar these are to those formed during irradiation. In this work atom probe tomography, combined with detailed structural assessments of the structure of solute clusters, is used to address this issue. A series of thermal ageing heat treatments has been performed on several high- and low-Ni RPV welds to produce 1-4 nm diameter solute clusters. The same materials have also been neutron irradiated. The results show that CuMnNiSi enriched clusters formed during thermal ageing have, on average, higher Cu contents and lower Mn, Ni and Si contents than those found in irradiation-induced clusters. The effect of increasing bulk Ni is to encourage the formation of clusters with significantly higher Ni content, slightly higher Mn and Si contents and significantly lower Cu contents. At very high doses and dose rates MnNiSi enriched clusters can form even in high-Cu welds. Despite differences in the compositions of individual clusters formed during irradiation and during thermal ageing, clusters in both exhibit similar structure. In particular, well developed clusters in both materials have Cu-enriched cores whose peripheries are enriched in Ni, Mn and, in most cases, Si.

A sensitivity analysis of the maximum separation method for the characterisation of solute clusters.

Variants of the maximum separation method have become the de-facto methodologies for the characterisation of nanometre scale clusters in atom probe tomography (APT) data obtained from dilute solid solutions. All variants rely on a number of parameters and it is well known that the precise values for these parameters strongly influence estimates of cluster size and number density. Quantitative analyses require an improved understanding of the inter-relationship between user-defined parameters, experimental parameters such as detection efficiency and the resultant parameterisation of the microstructure. A series of simulations has been performed to generate clusters with a range of compositions (50-100%) and diameters (1.5-2.5 nm) in a dilute solid solution. The data were degraded to simulate the effects of the finite detection efficiencies and positioning uncertainties associated with the ECOPoSAP and LEAP-3000X HR. An extensive analysis of each resultant dataset, using a range of values for the maximum separation parameters was then performed. Optimum values for each material condition were identified and it is shown that it is possible to characterise cluster size, number density and matrix chemistry. However, accurate estimates of cluster compositions are more difficult and absolute measurements must be treated with caution. Furthermore, it is shown that D(MAX) must increase with decreasing detection efficiency and consequently clusters of a specific size will appear slightly larger in atom probes with a lower detection efficiency.

Atom probe tomography of reactor pressure vessel steels: an analysis of data integrity.

In this work, the importance of optimising experimental conditions for the analysis of reactor pressure vessel (RPV) steels using atom probe tomography is explored. The quality of the resultant atom probe data is assessed in terms of detection efficiency, noise levels and mass resolution. It is demonstrated that artefacts can exist even when experimental conditions have been optimised. In particular, it is shown that surface diffusion of some minority species, including P and Si, to major poles prior to field evaporation can be an issue. The effects were most noticeable during laser pulsing. The impact of surface migration on the characterisation of dislocations and grain boundaries is assessed. The importance of selecting appropriate regions of the reconstructed data for subsequent re-analysis is emphasised.

Statistical analysis of atom probe data: detecting the early stages of solute clustering and/or co-segregation.

Statistical analysis of atom probe data has improved dramatically in the last decade and it is now possible to determine the size, the number density and the composition of individual clusters or precipitates such as those formed in reactor pressure vessel (RPV) steels during irradiation. However, the characterisation of the onset of clustering or co-segregation is more difficult and has traditionally focused on the use of composition frequency distributions (for detecting clustering) and contingency tables (for detecting co-segregation). In this work, the authors investigate the possibility of directly examining the neighbourhood of each individual solute atom as a means of identifying the onset of solute clustering and/or co-segregation. The methodology involves comparing the mean observed composition around a particular type of solute with that expected from the overall composition of the material. The methodology has been applied to atom probe data obtained from several irradiated RPV steels. The results show that the new approach is more sensitive to fine scale clustering and co-segregation than that achievable using composition frequency distribution and contingency table analyses.

Characterisation of the early stages of solute clustering in 1Ni-1.3Mn welds containing Cu.

Microstructural characterisation of neutron irradiated low alloy steels is important for developing mechanistic understanding of irradiation embrittlement. This work is focused on the early stages of irradiation-induced clustering in a low Cu (0.03wt%), high Ni ( approximately 1wt%) weld. The weld was irradiated at a very high dose rate and then examined by atom probe (energy-compensated position-sensitive atom probe (ECOPoSAP) and local electrode atom probe (LEAP)) with supporting microstructural information obtained by small angle neutron scattering (SANS) and positron annihilation (PALA). It was demonstrated that extreme care must be taken optimising parameters used to characterise the extent of clustering. This is particularly important during the early stages of irradiation-damage when the clusters are poorly defined and significant compositional variations are present in what is traditionally described as matrix. Analysis of the irradiated materials showed increasing clustering of Cu, Mn, Ni and Si with dose. In the low Cu steel the results showed that initially the irradiation damage results in clustering of Mn, Ni and Si, but at very high doses, at very high dose rates, redistribution of Si is significantly more advanced than that for Mn and Ni.

Olfactory performance during childhood. II. Developing a discrimination task for children.

OBJECTIVES: (1) To create a match-to-sample odorant discrimination task (MODT) for children and adolescents; (2) to assess whether nonolfactory factors affect olfactory performance more on an identification task than on the MODT; (3) to evaluate subjects with olfactory dysfunction; and (4) to create age-appropriate sets of odorants for use in the MODT format to test children of different ages. STUDY DESIGN: We tested 75 normal children, aged 2 to 18 years, and 17 other subjects, aged 7 to 53 years, with known or suspected olfactory dysfunction, with the MODT. We compared the age trends in variability of scores on the MODT with those on an odorant identification task, using a weighted linear regression analysis. RESULTS: The MODT was useful in children aged 5 years and older, but not generally in the 2- to 4-year-old children. There was an appreciable age trend in the variability of the scores on the identification task but not on the MODT. Mean MODT scores for subjects with suspected or known olfactory dysfunction were far below average. Finally, we created four sets of odorants that will likely be sensitive to age-specific changes in olfactory performance. CONCLUSIONS: The MODT appears to be a suitable test instrument to assess olfaction in children aged 5 and older and is less likely to be influenced by nonolfactory factors than an identification task. According to our preliminary results, it is likely that the MODT will allow us to detect olfactory deficits in children of many ages.

Constitutive over-expression of transforming growth factor-alpha in rat liver epithelial cells leads to increased cell cycling without transformation.

Over-expression of transforming growth factor-alpha (TGF-alpha) is consistently seen in spontaneous transformants of rat liver derived epithelial cells (RLE phi 13) and has been implicated in the transformation of other cultured cells. We have constitutively over-expressed TGF-alpha in RLE phi 13 cells, which are known to express epidermal growth factor receptors, to determine if TGF-alpha over-expression plays a role in transformation or differentiation, or both, of these cells. Early passage RLE phi 13 cells were infected with a replication-defective murine retrovirus that expresses both the full length coding sequence for human TGF-alpha and the neomycin-resistance gene. Integration of the transcriptionally active provirus and expression of TGF-alpha mRNA were confirmed. Neither morphologic transformation nor molecular evidence for differentiation was noted in TGF-alpha-producing clones. However, these clones did exhibit an accelerated growth rate, increased expression of several cell cycle related genes including mitotic cyclic B1, proliferating cell nuclear antigen, c-myc, and p53 as well as increased expression of the preneoplastic marker enzyme, glutathione-S-transferase. This suggests that over-expression of TGF-alpha results in increased cell cycling, and that subsequent events must be necessary for cellular transformation or differentiation or both.

Brain microvascular smooth muscle expresses class II antigens.

Mouse (BALB/c) splenic lymphocytes co-cultured in vitro with syngeneic brain-derived microvascular smooth muscle (SM) proliferate and become activated. After subsequent transfer of the activated lymphocytes to a syngeneic host, a vasculitis develops in the host. Investigation of the possible antigen-presenting properties of the cultured SM has resulted in the demonstration of class II (Ia) antigens on the SM. Fluorescence-activated cell sorter analysis has shown that an average of 31% of unstimulated SM cells in culture were positive when stained with an anti-IE of the appropriate haplotype (H2d), and an average of 20% were positive with an anti-IA of the H2d haplotype. Controls consisting of irrelevant antibodies of the same isotype, as well as an anti-IA of the H2s haplotype, were negative. In contrast, BALB/c-derived brain microvascular endothelial cells showed considerably less class II antigen expression (7% for both IA and IE).

The physiology of B cells as studied with tumor models.

B-cell tumors have been extraordinary sources of information about antibodies, their genes, and the cells that express them. An important principle that has emerged from the study of lymphoid tumors is that the long-held view that malignant lymphoid cells are "frozen" at a fixed point in differentiation is not generally valid. Presentation of immunoregulatory signals to transformed B cells can profoundly influence their proliferation, morphology, differentiation, gene expression, and immunoglobulin synthesis. In addition to their responsiveness to immunoregulatory signals, some tumors of B lineage elaborate immunoregulatory signals. Until quite recently B-cell tumors were used primarily as monoclonal sources of molecules of immunological interest. While they continue to be important sources of receptors, growth and differentiation factors, differentiation antigens, and immunoregulatory factors, they are being used with increasing frequency to define the molecular events that occur in B cells subsequent to receipt of an immunoregulatory signal. While the use of tumor cells as models of normal cells is often viewed with some skepticism, it is difficult to find examples wherein tumors have been misleading. Quite to the contrary, B-cell tumors have regularly provided powerful tools for dissecting the molecular events that underlie B-cell development, function, and regulation.

Structure and position of a complex chromosomal aberration in bacteriophage P2.

The P2 phage mutation vir56, like the previously studied vir22, is the result of an unequal replacement of a chromosome segment with non-homologous DNA. The end positions of the replacements are essentially the same in the two mutants, whereas the lengths of the replacements are quite different. A third chromosomal aberration, del3, has similar structure and position. These results strengthen the suggestion that the left ends of these three aberrations coincide with the point of exchange in integrative recombination.

Kinetics of viral deoxyribonucleic acid, protein, and infectious particle production and alterations in host macromolecular syntheses in equine abortion (herpes) virus-infected cells.

Infection of exponential-phase suspension cultures of mouse fibroblast cells (L-M) with equine abortion virus (EAV) resulted in inhibition of cell growth and marked alterations in host metabolic processes. The synthesis of deoxyribonucleic acid (DNA) and ribonucleic acid was inhibited within 4 hr after infection and was suppressed by more than 90% by the time of maximal virus replication (14 to 18 hr). The overall rate of protein synthesis, however, was similar in uninfected and virus-producing cells as determined by measurements of net protein and isotope incorporation. The time course of viral DNA and protein synthesis and assembly into mature virus was determined with the inhibitors 5-fluorodeoxyuridine (FUdR) and cycloheximide, respectively. Thus, viral DNA synthesis was essentially completed at 14 hr, and viral protein and infectious virus synthesis was completed at 18 hr. Although the number of plaque-forming units (PFU) produced by FUdR-treated cells (10(3) to 10(4) PFU/ml) was at least 3 logs less than that produced by untreated cells, the yield of physical particles (as determined by electron microscopy) was approximately the same at 30 hr after infection. Besides being relatively non-infective, the particles produced in FUdR-treated cells appeared morphologically incomplete as they contained little or no nucleoid material.

Fine structure of quiescent and germinating aeciospores of Cronartium fusiforme.

Aeciospores of the long-cycle heteroecious rust fungus, Cronartium fusiforme, were found to have an extremely thick cell wall with striking spicules protruding from it. The wall was readily degraded by commercial chitinase, but spicules were unaffected. Quiescent spores contained two nuclei with distinct nuclear membranes possessing many pores. Numerous membrane-bounded lipid bodies were found both in wild-type orange and in white mutant aeciospores. An abundance of irregularly ovoid mitochondria was present in quiescent spores. After glutaraldehydeosmium fixation, the surface of the mitochondria appeared to be covered with ribosomes or microtubules in a paracrystalline array, whereas after permanganate fixation only smooth outer mitochondrial membranes were noted. The latter fixative revealed abundant vesicular endoplasmic reticulum in the spore. Spores incubated at 20 C on agar produced one to five distinct germ tubes within 65 to 180 min. These thin-walled tubes exhibited varying degrees of branching, and reached a total hyphal length of 300 to 500 mu prior to rupturing. Emergence of germ tubes took place through a pore in the spore wall and appeared to be mainly a physical flowing of cytoplasm from the spore into the germ tube without division of nuclei or other cell organelles. On completion of germination, the protoplasm of the germ tube contained both nuclei and nearly all of the other spore contents. Mitochondria had smooth outer membranes, were greatly elongated, and possessed distinct longitudinal cristae. A limited amount of rough endoplasmic reticulum was arranged parallel to the germ tube wall. Other organelles seen in germ tubes were lipid bodies, concentric membrane figures, and numerous ribosomes. Lipid bodies appeared smaller and fewer in number than in quiescent spores.

Ultrastructure of basidiospores and mycelium of Lenzites saepiaria.

Hyde, James M. (University of Mississippi Medical School, Jackson), and Charles H. Walkinshaw. Ultrastructure of basidiospores and mycelium of Lenzites saepiaria. J. Bacteriol. 92:1218-1227. 1966.-Ungerminated and germinated basidiospores and 2-day-old mycelial cultures of Lenzites saepiaria were similar in their fine structure. Fixation with glutaraldehyde, followed by osmium tetroxide, was far superior to permanganate. Cell organelles were seen in cytoplasm of spores and hyphae, and clamp connections were abundant in hyphal elements. Numerous lomasomes, vesicular bodies, and complex concentric membranes occurred in the cytoplasm and were often associated with the cell membrane or the dolipore membrane (parenthesome) of the septum. Endoplasmic reticulum was not found, but numerous ribosomes were seen; polyribosome groupings were frequently noted. The nucleus was bounded by a double membrane which contained few pores. Germinating spores exhibited one or more large osmiophilic bodies in association with a vacuole and membranous elements. Other than possessing a thin wall, the emerging germ tube was similar in structure to the parent spore.

Physicochemical properties of fowlpox virus deoxyribonucleic acid and its anomalous infectious behavior.

Randall, Charles C. (University of Mississippi, Jackson), Lanelle G. Gafford, Richard L. Soehner, and James M. Hyde. Physicochemical properties of fowlpox virus deoxyribonucleic acid and its anomalous infectious behavior. J. Bacteriol. 91:95-100. 1966.-Deoxyribonucleic acid (DNA) was extracted from fowlpox virus-infected tissue, purified inclusions, and purified virus by five variations of detergent and phenol methods. Phenol methods gave a poor yield, whereas detergent techniques extracted up to 78% of the DNA. The buoyant density was 1.695 g/ml, and the melting temperature in 7.2 m NaClO(4) was 39 C, both approximately equivalent to a guanine plus cytosine content of 35 moles per cent. Further proof of the double-stranded nature of the DNA was shown by the characteristic behavior toward deoxyribonuclease, formaldehyde, and heat. Infectious DNA was obtained by the various methods described, but this manifestation of biological activity was capricious and for unknown reasons was often not evident. The infectivity could not be related quantitatively to the amount of DNA employed. Furthermore, the infectious nature of fowlpox virus DNA was demonstrable only when the route of infection was the chorioallantoic membrane. In contrast, whole virus infected both membrane and chick skin with equal efficiency.