bluuebunny/crossref_metadata_2025 huggingface dataset:
Soil-water characteristic curve and consolidation behavior for a compacted silt
Authors: Thu Trinh Minh , Rahardjo Harianto , Leong Eng-Choon
Published: June 2007

Measurement of the soil-water characteristic curve (SWCC) in the laboratory is commonly conducted under zero confining pressure. However, in the field, the soil is under a confining stress. Therefore, it is important to study the effects of the confining stress on SWCC. In addition, the consolidation curve is normally generated under saturated conditions. However, the soil above the water table is usually unsaturated. Hence, it is also necessary to investigate the effects of matric suction on the characteristics of the consolidation curves. This paper presents the SWCCs under different net confining stresses and the isotropic consolidation curves under different matric suctions that describe the volume change characteristics of unsaturated soils with respect to stress state variables, net normal stress, and matric suction. A series of SWCCs was determined for statically compacted silt specimens in a triaxial cell apparatus under different net confining stresses. Isotropic consolidation tests under different matric suctions were also carried out. The results of the SWCC tests show that the air-entry value increased with increasing net confining stress. The yield points (i.e., yield suction, s0) obtained from the SWCC tests also increased with increasing net confining stress. The results of isotropic consolidation tests indicate the strong influence of matric suction on compressibility and stiffness of the compacted silt specimens.Key words: soil-water characteristic curve, isotropic consolidation, pore-water pressure, volume change, NTU mini suction probe, matric suction.
Three-dimensional stability evaluation of a preexisting landslide with multiple sliding directions by the strength-reduction technique
Authors: Deng J H , Tham L G , Lee C F , Yang Z Y
Published: June 2007

Landslide 1 is located on the right bank of the downstream side of the dam for Hongjiadu Hydropower Station in Guizhou Province, China. In view of its close proximity to important workings, the stability of the slide is of great concern. Ground investigation showed that the slide is potentially unstable and has two sliding directions. Reinforcement mainly by piles was proposed as the stabilization measure, but two key questions were raised during the design of the piles. (1) How could the stability of the slope be evaluated, as there are two major sliding directions? (2) How could the shear strength parameters of the slip band be determined, as it contains a significant proportion of coarse particles? In this paper, we demonstrate how these issues can be addressed by the strength-reduction technique. First, the strength parameters of the slip band are back-analyzed by assuming a factor of safety of unity. Second, the major sliding directions are determined by carrying out finite difference analyses. Third, two proposed stabilization schemes are evaluated and compared to demonstrate the versatility of the technique. The present study has demonstrated that the strength-reduction technique is a powerful tool for analyzing such problems.Key words: landslide, reinforcement, piles, stability evaluation, strength-reduction technique.
Measuring the onset of mine tailings erosion
Authors: Haneef-Mian M , Yanful Ernest K , Martinuzzi Robert
Published: June 2007

The present study gives details of a methodology for estimating the critical shear stress for erosion of mine tailings and other naturally occurring cohesive sediments. Erosion of a cohesive sediments bed occurs when the critical shear stress is exceeded to break the interparticle bond. Experiments were conducted in a 30 cm diameter laboratory column and calibrated using laser Doppler anemometry. The results showed that the erosion pattern of mine tailings particles was similar to those of fine-grained cohesive sediments. A power-law relation of the form E = α[(τ – τcr)/τcr]nis suggested for mine tailings, where E is the erosion rate, α is a coefficient, τ is the shear stress, τcris the critical shear stress, and n is an exponent. The computed values of α, n, and τcrin the power-law equation were found to be comparable to values derived from experiments in a rotating circular flume. The derived expression for rate of erosion may be incorporated in resuspension and transport models for fine mine tailings of a similar nature.Key words: mine tailings, laser Doppler velocimetry, wall shear stresses, critical shear stress for erosion, erosion – shear stress relationship.
Field experimental cells to evaluate the hydrogeological behaviour of oxygen barriers made of silty materials
Authors: Bussière Bruno , Aubertin Michel , Mbonimpa Mamert , Molson John W , Chapuis Robert P
Published: June 2007

An in situ study was initiated to evaluate the efficiency of silts as moisture-retaining material in covers with capillary barrier effects (CCBEs) that aim at controlling the production of acid mine drainage (AMD). This investigation included the construction of four experimental cells covered by different layered cover configurations. Each CCBE was instrumented to monitor its hydrogeological behaviour. The degree of saturation (Sr) in the moisture-retaining (silty) materials was usually maintained above 85%, while Sr in the two sand layers placed above and below the silts was generally lower than 30%–50% (typical hydrogeological behaviour of efficient CCBEs). Suction measurements in the different layers of the four CCBEs were consistent with the observed Sr measurements during the four years of monitoring. The suctions measured in the moisture-retaining materials were lower than their respective air-entry values (AEV), while suctions largely exceeded the AEV of the sand in the capillary break layers. The oxygen flux calculations confirm that in each of the four CCBEs the flux through the CCBEs remained low for the entire duration of the experiment, hence limiting the generation of AMD. This study also confirmed the ability of silty materials to act as moisture retaining in a CCBE. Key words: cover with capillary barrier effects, acid mine drainage, experimental cells, in situ monitoring, unsaturated flow, diffusion, oxygen flux.
Submarine slope failure due to gas hydrate dissociation: a preliminary quantification
Authors: Nixon M F , Grozic J LH
Published: June 2007

Gas hydrates are icelike compounds composed of water and methane gas in very compact form. There is substantial evidence from case histories that links gas hydrate dissociation to submarine slope failures and other geohazards. Theoretical analyses have also shown that upon dissociation gas hydrates will cause an increase in fluid pressure and a reduction in effective stress and thus result in loss of the soil strength. This paper presents a preliminary quantification of the effects of gas hydrate dissociation through development of a pore-pressure model that was incorporated into one- and two-dimensional slope stability analyses. The ensuing numerical study investigated submarine slope stability through parametric studies and application to two important case histories and found that dissociation of even small amounts of hydrate can have a significant destabilizing effect. Yet whether gas hydrate dissociation can alone cause large-scale slope failures has still to be demonstrated as there are often many destabilizing processes; however, this research highlights the importance of assessing the effects of gas hydrate dissociation on the behaviour of submarine slopes.Key words: gas hydrates, slope stability, marine, offshore, methane gas, instability.
The earth pressure behind full-height frame integral abutments supporting granular fill
Authors: Xu Ming , Clayton Chris RI , Bloodworth Alan G
Published: June 2007

Compared with conventional bridges, integral bridges have no bearings or joints between the deck and abutments and thus can significantly reduce maintenance requirements and costs over the bridge's lifetime. However, there is uncertainty about the ultimate magnitude of the lateral earth pressure behind such abutments, as they are forced to move with the deck length change caused, for example, by daily and annual variations in the effective bridge temperature. This research investigated the earth pressure that would be expected to occur behind full-height frame integral abutments backfilled by granular materials. Radial strain-controlled cyclic stress path testing has been conducted on coarse sand specimens and a glass ballotini specimen. The results suggest that for integral abutments retaining uniform coarse sand, the lateral earth pressure will experience systematic increases for almost all cyclic strain levels, eventually reaching states of stress close to both active and passive. The mechanism of the buildup of lateral stress is explored, and it appears to be associated with nonspherical granular particle shape. The implications for frame integral abutment design are discussed.Key words: integral abutments, granular, particle shape, earth pressure, stiffness.
The horizontal-moment capacity of embedded foundations in undrained soil
Authors: Yun Gijae , Bransby M Fraser
Published: June 2007

The bearing capacity of embedded foundations subject to combined horizontal and moment loading in undrained soils is of interest particularly to the offshore geotechnical engineer. These loadings can be experienced by monopod foundations and offshore manifolds and often take the form of a horizontal load applied at a lever arm height above the foundation base. Previous design methods using the failure envelope approach have suggested that peak moment capacity of a foundation is mobilized with positive horizontal loads, but that peak horizontal load is mobilized with no additional moment loading. This paper reports numerical work specifically investigating the effects of the embedment ratio on the horizontal-moment foundation capacity under no vertical load in both uniform strength and "normally consolidated" undrained soil. It is shown that the embedment ratio significantly affects the eccentricity of the M–H failure envelope so that the shape varies when normalized by the pure moment (M0) and pure horizontal load (H0) capacity. The reasons for the eccentricity of the failure envelopes are understood by investigating soil deformation mechanisms calculated during FE analysis and by accompanying upper bound plasticity analysis. This mechanistic understanding is used to suggest load reference point translations that ease curve-fitting of the complex failure envelopes for use in design.Key words: bearing capacity, foundations, numerical modelling and analysis, plasticity.
Evaluation of the transitional inelastic behaviour of unsaturated clay–sand mixtures
Authors: Blatz James , Anderson David E.S , Siemens Greg
Published: June 2007

This paper examines and compares the mechanical behaviour of two different unsaturated clay mixtures comprised of bentonite clay (Saskatchewan or Wyoming) and quartz sand. The two mixtures have been proposed as compacted barrier materials for reducing groundwater flow in the vicinity of waste disposal repositories. Triaxial specimens were compacted to consistent properties, and then specified suction conditions were applied to the specimens using the vapour equilibrium technique. Following equilibrium at the specified initial suction, specimens were subjected to isotropic and shear loading in a conventional triaxial cell to measure the mechanical response under selected stress paths. The results are interpreted in terms of the yield, strength, and stiffness behaviour at the various suction levels. Results suggest that the clay component of the mixture dominates the behaviour at suctions less than approximately 30 MPa, and the sand component dominates the behaviour above approximately 30 MPa. The transition from clay- to sand-dominated behaviour is attributed to volume strain during application of the initial suction bringing the sand particles into contact. The discussion highlights how the results can be used to modify constitutive models to incorporate the transitional behaviour in numerical modeling.Key words: inelastic, yielding, unsaturated, stress–strain, triaxial testing.
Modelling the influence of stress–strain history on the initial shear stiffness of an unsaturated compacted silt
Authors: Vassallo Roberto , Mancuso Claudio , Vinale Filippo
Published: June 2007

This is the follow-up paper to Vassallo et al. (2007), which discussed the experimentally observed small-strain behaviour of an unsaturated compacted silt. The influence that suction and, more in general, mean net stress – suction history has on the initial shear stiffness was analysed and ascribed to the accumulation of irreversible volumetric strains. In this study, a model able to predict the observed behaviour is proposed, based on classical unsaturated soil volumetric hardening elastoplastic formulations. Starting from the interpretation of the results relative to simple stress paths, such as preliminary equalization and loading–unloading compression, the results of "complex" stress paths, such as those including drying–wetting cycles, have subsequently been modelled by introducing some additional parameters.Key words: unsaturated, compacted, small strain, stiffness, volumetric behaviour, stress history.
Post-closure investigation of engineered test covers on acid-generating waste rock at Whistle Mine, Ontario
Authors: Adu-Wusu Celestina , Yanful Ernest K
Published: June 2007

Three engineered test covers were installed at the Whistle Mine test site, near Capreol, Ontario, and monitored over 4 years. Each cover was a two-layer system consisting of a 0.90 m noncompacted gravelly sand overlying a barrier layer and a 6.10 m waste rock platform. The barriers consisted of a 92% sand – 8% bentonite mixture, a sandy silt, and a geosynthetic clay liner (GCL). A control plot (waste rock without cover) was also monitored. Water percolation through the covers was 61% for the sandy silt cover, 24% for the sand–bentonite mixture, 12% for the GCL, and 58% for the control. Post-monitoring excavation and examination of the test plots indicated that defects in the placement of the barrier layers, coupled with freeze–thaw effects, likely contributed to the unexpected high water percolation values observed. The results suggest that lapses in construction of soil barriers, such as lack of proper installation and quality control, can compromise cover effectiveness.Key words: mine waste, remediation, soil covers, post closure, investigations, water balance.
A procedure for the design of protective filters
Authors: Babu G L. Sivakumar , Srivastava Amit
Published: June 2007

Current procedures for the design of filters to protect the soil from erosion and piping are solely based on particle-size distribution (PSD) curves of the filter medium and base soil. In the present work, an analytical solution is presented in a single derivation for the design of the filter, which takes into account factors like pore size, permeability, and factor of safety against soil boiling conditions. The solution obtained from the proposed analytical procedure provides insight into the role of seepage velocity, relative density of the filter material, and the ratio of filter density to base soil density on the performance of the filter.Key words: filter, erosion, particle-size distribution, permeability, seepage velocity, relative density.
Bcl10 mediates LPS-induced activation of NF-κB and IL-8 in human intestinal epithelial cells
Authors: Bhattacharyya Sumit , Borthakur Alip , Pant Nitika , Dudeja Pradeep K. , Tobacman Joanne K.
Published: June 2007

Lipopolysaccharide (LPS) is recognized as an inducer of the inflammatory response associated with gram-negative sepsis and systemic inflammatory response syndrome. LPS induction proceeds through Toll-like receptor (TLR) in immune cells and intestinal epithelial cells (IEC). This report presents the first identification of Bcl10 (B-cell CLL/lymphoma 10) as a mediator of the LPS-induced activation of IL-8 in human IEC. Bcl10 is a caspase-recruitment domain-containing protein, associated with constitutive activation of NF-κB in MALT (mucosa-associated lymphoid tissue) lymphomas. The normal human IEC line NCM460, normal primary human colonocytes, and ex vivo human colonic tissue were exposed to 10 ng/ml of LPS for 2–6 h. Effects on Bcl10, phospho-IκBα, NF-κB, and IL-8 were determined by Western blot, ELISA, immunohistochemistry, and confocal microscopy. Effects of Bcl10 silencing by small-interfering RNA (siRNA), TLR4 blocking antibody, TLR4 silencing by siRNA, and an IL-1 receptor-associated kinase (IRAK)-1/4 inhibitor on LPS-induced activation were examined. Following Bcl10 silencing, LPS-induced increases in NF-κB, IκBα, and IL-8 were significantly reduced ( P < 0.001). Increasing concentrations of LPS were associated with higher concentrations of Bcl10 protein when quantified by ELISA, and the association between LPS exposure and increased Bcl10 was also demonstrated by Western blot, immunohistochemistry, and confocal microscopy. Exposure to TLR4 antibody, TLR4 siRNA, or an IRAK-1/4 inhibitor eliminated the LPS-induced increases in Bcl10, NF-κB, and IL-8. Identification of Bcl10 as a mediator of LPS-induced activation of NF-κB and IL-8 in normal human IEC provides new insight into mechanisms of epithelial inflammation and new opportunities for therapeutic intervention.
Three distinct phases of VF during global ischemia in the isolated blood-perfused pig heart
Authors: Huizar Jose F. , Warren Mark D. , Shvedko Alexander G. , Kalifa Jérôme , Moreno Javier , Mironov Sergey , Jalife José , Zaitsev Alexey V.
Published: June 2007

Changes in ventricular fibrillation (VF) organization occurring after the onset of global ischemia are relevant to defibrillation and survival but remain poorly understood. We hypothesized that ischemia-specific dynamic instability of the action potential (AP) causes a loss of spatiotemporal periodicity of propagation and broadening of the electrocardiogram (ECG) frequency spectrum during VF in the ischemic myocardium. We recorded voltage-sensitive fluorescence of di-4-ANEPPS (anterior left ventricle, 35 × 35 mm, 64 × 64 pixels) and the volume-conducted ECG in six blood-perfused hearts during 10 min of VF and global ischemia. We used coefficient of variation (CV) to estimate variability of AP amplitude, AP duration, and diastolic interval (CV-APA, CV-APD, and CV-DI, respectively). We computed excitation median frequency (Median_F), spectral width of the AP and ECG (SpW-AP and SpW-ECG, respectively), wavebreak incidence (WBI), and recurrence of propagation direction (RPD). We found three distinct phases of local VF dynamics: “relatively periodic” (≤1 min, high Median_F, moderate AP variability, high WBI, low RPD), “highly periodic” (1–2 min, reduced Median_F, low AP variability, low WBI, high RPD), and “aperiodic” (3–10 min, low Median_F, high AP variability, high WBI, low RPD). In one experiment, spontaneous conversion from the aperiodic to the highly periodic phase occurred after 5 min of ischemia. The SpW-ECG was correlated with SpW-AP, CV-APD, and CV-APA. We conclude that 1) at least three distinct phases of VF dynamics are present in our model, and 2) the newly described aperiodic phase is related to ischemia-specific dynamic instability of the AP shape, which underlies broadening of the ECG spectrum during VF evolution.
Parvalbumin isoforms differentially accelerate cardiac myocyte relaxation kinetics in an animal model of diastolic dysfunction
Authors: Rodenbaugh David W. , Wang Wang , Davis Jennifer , Edwards Terri , Potter James D. , Metzger Joseph M.
Published: June 2007

The cytosolic Ca2+/Mg2+-binding protein α-parvalbumin (α-Parv) has been shown to accelerate cardiac relaxation; however, beyond an optimal concentration range, α-Parv can also diminish contractility. Mathematical modeling suggests that increasing Parv's Mg2+ affinity may lower the effective concentration of Parv ([Parv]) to speed relaxation and, thus, limit Parv-mediated depressed contraction. Naturally occurring α/β-Parv isoforms show divergence in amino acid primary structure (57% homology) and cation-binding affinities, with β-Parv having an estimated 16% greater Mg2+ affinity and ∼200% greater Ca2+ affinity than α-Parv. We tested the hypothesis that, at the same or lower estimated [Parv], mechanical relaxation rate would be more significantly accelerated by β-Parv than by α-Parv. Dahl salt-sensitive (DS) rats were used as an experimental model of diastolic dysfunction. Relaxation properties were significantly slowed in adult cardiac myocytes isolated from DS rats compared with controls: time from peak contraction to 50% relaxation was 57 ± 2 vs. 49 ± 2 (SE) ms ( P < 0.05), validating this model system. DS cardiac myocytes were subsequently transduced with α- or β-Parv adenoviral vectors. Upon Parv gene transfer, β-Parv caused significantly faster relaxation than α-Parv ( P < 0.05), even though estimated [β-Parv] was ∼10% of [α-Parv]. This comparative analysis showing distinct functional outcomes raises the prospect of utilizing naturally occurring Parv variants to address disease-associated slowed cardiac relaxation.
Dynamic changes in nitric oxide and mitochondrial oxidative stress with site-dependent differential tissue response during anoxic preconditioning in rat heart
Authors: Cuong Dang Van , Warda Mohamad , Kim Nari , Park Won Sun , Ko Jae Hong , Kim Euiyong , Han Jin
Published: June 2007

In this study, dynamic changes in nitric oxide (NO) and mitochondrial superoxide (O2•−) were examined during anoxic preconditioning (AP) in rat heart model. AP and anoxia-reoxygenation (A/R) were performed on isolated hearts and single cardiomyocytes. The cellular insult in the form of infarct size and DNA damage were localized and correlated with NO synthases (endothelial and inducible) expression levels. The results showed that endocardium was the most affected region in AP groups, whereas the larger area of infarct was confined to mid- and epicardium in the A/R group. Interestingly, a high-level expression of immunofluorescent NO synthases was restricted to viable areas in the AP. In contrast to the gradual increase in O2•− level that occurred in the AP group, a sudden massive increase in its level was demonstrated at the onset of reoxygenation in the A/R group. The observed increase in NO production during reoxygenation in the AP group was attenuated by inducible NO synthase inhibitor. The study revealed, on a real-time basis, the role played by preconditioning for modulating NO and O2•− levels on behalf of cell survival. The results afford a better understanding of cardiac-adapting mechanism during AP and the role of inducible NO synthase in this important phenomenon.
Involvement of cell surface ATP synthase in flow-induced ATP release by vascular endothelial cells
Authors: Yamamoto Kimiko , Shimizu Nobutaka , Obi Syotaro , Kumagaya Shinichiro , Taketani Yutaka , Kamiya Akira , Ando Joji
Published: June 2007

Endothelial cells (ECs) release ATP in response to shear stress, a mechanical force generated by blood flow, and the ATP released modulates EC functions through activation of purinoceptors. The molecular mechanism of the shear stress-induced ATP release, however, has not been fully elucidated. In this study, we have demonstrated that cell surface ATP synthase is involved in shear stress-induced ATP release. Immunofluorescence staining of human pulmonary arterial ECs (HPAECs) showed that cell surface ATP synthase is distributed in lipid rafts and co-localized with caveolin-1, a marker protein of caveolae. Immunoprecipitation indicated that cell surface ATP synthase and caveolin-1 are physically associated. Measurement of the extracellular metabolism of [3H]ADP confirmed that cell surface ATP synthase is active in ATP generation. When exposed to shear stress, HPAECs released ATP in a dose-dependent manner, and the ATP release was markedly suppressed by the membrane-impermeable ATP synthase inhibitors angiostatin and piceatannol and by an anti-ATP synthase antibody. Depletion of plasma membrane cholesterol with methyl-β-cyclodextrin (MβCD) disrupted lipid rafts and abolished co-localization of ATP synthase with caveolin-1, which resulted in a marked reduction in shear stress-induced ATP release. Pretreatment of the cells with cholesterol prevented these effects of MβCD. Downregulation of caveolin-1 expression by transfection of caveolin-1 siRNA also markedly suppressed ATP-releasing responses to shear stress. Neither MβCD, MβCD plus cholesterol, nor caveolin-1 siRNA had any effect on the amount of cell surface ATP synthase. These results suggest that the localization and targeting of ATP synthase to caveolae/lipid rafts is critical for shear stress-induced ATP release by HPAECs.
Energetic cost of synthesizing proteins in Antarctic limpet,Nacella concinna(Strebel, 1908), is not temperature dependent
Authors: Bowgen Andrew D. , Fraser Keiron P. P. , Peck Lloyd S. , Clarke Andrew
Published: June 2007

The energetic cost of protein synthesis is thought to account for a significant proportion of total metabolism. However, attempts to estimate the energetic cost of synthesizing proteins have resulted in surprisingly variable results, particularly for the small number of polar organisms studied, where cost estimates vary by two orders of magnitude. Much of this variability is probably the result of differing methodologies and experimental designs. Here we have used two different, carefully validated methods to measure the costs of protein synthesis in Antarctic limpets. One method, which utilized a specific protein synthesis inhibitor, was used to measure the cost of protein synthesis at two temperatures to test the hypothesis that the cost of protein synthesis varies with temperature. The cost of protein synthesis measured using the inhibitor cycloheximide was 13.95 ± 0.77 μmol O2/mg protein, while correlation of absolute protein synthesis with oxygen consumption suggested the cost of protein synthesis was 19.58 μmol O2/mg protein. Water temperature did not alter the cost of protein synthesis in Nacella concinna (Student's t-test, P = 0.849, t = 0.19, df = 12). In a meta-analysis of literature values for the cost of protein synthesis there was no significant effect of temperature, but there was a significant relationship between the concentration of cycloheximide used to inhibit protein synthesis and the measured cost.
Studies of local and intermediate range structure in crystalline and amorphous materials at high pressure using high-energy X-rays
Authors: Ehm Lars , Antao Sytle M. , Chen Jiuhua , Locke Darren R. , Marc Michel F. , David Martin C. , Yu Tony , Parise John B. , Antao Sytle M. , Lee Peter L. , Chupas Peter J. , Shastri Sarvjit D. , Guo Quanzhong
Published: June 2007

The method of high-energy total elastic X-ray scattering to determine the atomic structure of nanocrystalline, highly disordered, and amorphous materials is presented. The current state of the technique, its potential, and limitations are discussed with two successful studies on the pressure induced phase transition in mackinawite (FeS) and the high-pressure behavior of liquid gallium.
Mechanics of microelectronics structures as revealed by X-ray diffraction
Authors: Murray Conal E. , Yan H. , Noyan I. C.
Published: June 2007

The presence of strain distributions within semiconductor features influences many aspects of their behavior. For example, microelectronic technology that incorporates strained silicon improves device performance by increasing carrier mobility in the Si channels. Because current semiconductor fabrication contains multiple levels of metallic and dielectric structures, an understanding of the mechanical response of the constituent elements is critical to the prediction of the overall device performance. In addition, the interaction of strain fields between adjacent structures becomes greater as feature sizes decrease and the corresponding feature density increases. The use of synchrotron-based X-ray methods allows one to determine the interaction between strained features and their environment at a submicron resolution. Real-space mapping of strain distributions in pseudomorphically strained, raised SiGe structures revealed that elastic relaxation extends approximately 20 times the feature thickness from their edges. X-ray topographic methods were also applied to map the substrate deformation induced by overlying SiGe features. A formulation based on the classical Ewald-von Laue theory of dynamical diffraction was derived to match the measured diffraction profiles.
Fundamental parameters analysis of RoHS elements in plastics
Authors: Elam W. T. , Shen Robert B. , Scruggs Bruce , Nicolosi Joseph A.
Published: June 2007

European Community Directive 2002∕95∕EC restricts the use of certain hazardous substances in electrical and electronic equipment. In particular, restrictions are placed on lead, mercury, cadmium, hexavalent chromium, and bromine (in polybrominated biphenyls or polybrominated diphenyl ethers). XRF is a convenient method for detecting the presence and measuring the amounts of these elements. Reliably quantifying all of these elements in plastics typically requires a large number of standards that are not yet readily available. Because of the light element matrix, using a “standardless” fundamental parameters method requires some reliance on the primary beam scatter, complicating the analysis algorithm and increasing the uncertainty. We have tested a simplified fundamental parameters method that determines the matrix via difference, requiring only one standard. The method was tested on a series of reference materials containing all of the regulated elements in a variety of plastic resins. One multi-element reference standard was used. It was necessary to include all of the additives in the specimens to achieve good quantitative accuracy. In addition, the scattered primary intensity was used in one set of tests to compensate for variations in specimen thickness. This thickness compensation was necessary to get acceptable results for Cd. Results were very promising, with average relative errors and relative standard deviations of about 10%.
DNA Methylation and Complete Transcriptional Silencing of Cancer Genes Persist after Depletion of EZH2
Authors: McGarvey Kelly M. , Greene Eriko , Fahrner Jill A. , Jenuwein Thomas , Baylin Stephen B.
Published: June 2007

Recent work suggests a link between the polycomb group protein EZH2 and mediation of gene silencing in association with maintenance of DNA methylation. However, we show that whereas basally expressed target cancer genes with minimal DNA methylation have increased transcription during EZH2 knockdown, densely DNA hypermethylated and silenced genes retain their methylation and remain transcriptionally silent. These results suggest that EZH2 can modulate transcription of basally expressed genes but not silent genes that are densely DNA methylated. [Cancer Res 2007;67(11):5097–102]
Endothelial Akt Signaling Is Rate-Limiting for Rapamycin Inhibition of Mouse Mammary Tumor Progression
Authors: Phung Thuy L. , Eyiah-Mensah Godfred , O'Donnell Rebekah K. , Bieniek Radoslaw , Shechter Sharon , Walsh Kenneth , Kuperwasser Charlotte , Benjamin Laura E.
Published: June 2007

Chronic activation of Akt signaling in the endothelium recapitulates the salient features of a tumor vasculature and can be inhibited by rapamycin, an inhibitor of mammalian target of rapamycin. This led to the hypothesis that the antitumor efficacy of rapamycin may be partially dependent on its ability to inhibit endothelial Akt signaling, making rapamycin an antiangiogenic agent and endothelial Akt pathway inhibitor. Dose-response studies with rapamycin showed that primary human endothelial cells and fibroblasts had a bimodal Akt response with effective reductions in phosphorylated Akt (pAkt) achieved at 10 ng/mL. In contrast, rapamycin increased pAkt levels in tumor cell lines. When tumor-bearing mice were treated with rapamycin doses comparable to those used clinically in transplant patients, we observed strong inhibition of mammary tumor growth. To test whether Akt activation in the endothelium was rate-limiting for this antitumor response, we engineered mouse mammary tumor virus–polyoma virus middle T antigen mice with endothelial cell–specific expression of constitutively activated Akt. We observed that the antitumor efficacy of rapamycin was reduced in the presence of elevated endothelial Akt activation. Just as we observed in MCF7 cells in vitro, rapamycin doses that were antiangiogenic resulted in increased pAkt levels in total mouse mammary tumor virus–polyoma virus middle T antigen tumor lysates, suggesting that tumor cells had an opposite Akt response following mammalian target of rapamycin inhibition compared with tumor endothelial cells. Together, these data support the hypothesis that endothelial Akt signaling in the tumor vasculature is an important target of the novel anticancer drug rapamycin. [Cancer Res 2007;67(11):5070–5]
Short Telomere Length and Breast Cancer Risk: A Study in Sister Sets
Authors: Shen Jing , Terry Mary Beth , Gurvich Irina , Liao Yuyan , Senie Ruby T. , Santella Regina M.
Published: June 2007

Telomeres consist of a tandem repeats of the sequence TTAGGG at the ends of chromosomes and play a key role in the maintenance of chromosomal stability. Previous studies indicated that short telomeres are associated with increased risk for human bladder, head and neck, lung, and renal cell cancer. We investigated the association between white blood cell telomere length and breast cancer risk among 268 family sets (287 breast cancer cases and 350 sister controls). Telomere length was assessed by quantitative PCR. The mean telomere length was shorter in cases (mean, 0.70; range, 0.03–1.95) than in unaffected control sisters (mean, 0.74; range, 0.03–2.29), but no significant difference was observed (P = 0.11). When subjects were categorized according to the median telomere length in controls (0.70), affected sisters had shorter telomeres compared with unaffected sisters after adjusting for age at blood donation and smoking status [odds ratio (OR), 1.3; 95% confidence interval (95% CI), 0.9–1.8], but the association was not statistically significant. The association by quartile of telomere length (Q4 shortest versus Q1 longest) also supported an increase in risk from shorter telomere length, although the association was not statistically significant (OR, 1.6; 95% CI, 0.9–2.7). This association was more pronounced among premenopausal women (OR, 2.1; 95% CI, 0.8–5.5) than postmenopausal women (OR, 1.3; 95% CI, 0.5–3.6 for Q4 versus Q1). If these associations are replicated in larger studies, they provide modest epidemiologic evidence that shortened telomere length may be associated with breast cancer risk. [Cancer Res 2007;67(11):5538–44]
Melanocyte Expression of Survivin Promotes Development and Metastasis of UV-Induced Melanoma in HGF-Transgenic Mice
Authors: Thomas Joshua , Liu Tong , Cotter Murray A. , Florell Scott R. , Robinette Kyle , Hanks Adrianne N. , Grossman Douglas
Published: June 2007

We previously found the apoptosis inhibitor Survivin to be expressed in melanocytic nevi and melanoma but not in normal melanocytes. To investigate the role of Survivin in melanoma development and progression, we examined the consequences of forced Survivin expression in melanocytes in vivo. Transgenic (Tg) mouse lines (Dct-Survivin) were generated with melanocyte-specific expression of Survivin, and melanocytes grown from Dct-Survivin mice expressed Survivin. Dct-Survivin melanocytes exhibited decreased susceptibility to UV-induced apoptosis but no difference in proliferative capacity compared with melanocytes derived from non-Tg littermates. Induction of nevi in Dct-Survivin and non-Tg mice by topical application of 7,12-dimethylbenz(a)anthracene did not reveal significant differences in lesion onset (median, 10 weeks) or density (4 lesions per mouse after 15 weeks). Dct-Survivin mice were bred with melanoma-prone MH19/HGF-B6 Tg mice, and all progeny expressing either individual, neither, or both (Survivin/HGF) transgenes were UV-treated as neonates and then monitored for 43 weeks. Melanocytes in neonatal Survivin+/HGF+ mouse skin were less susceptible to UV-induced apoptosis than those from Survivin−/HGF+ mice. Onset of melanocytic tumors was earlier (median, 18 versus 24 weeks; P = 0.01, log-rank test), and overall tumor density was greater (7.7 versus 5.2 tumors per mouse; P = 0.04) in Survivin+/HGF+ compared with Survivin−/HGF+ mice. Strikingly, melanomas arising in Survivin+/HGF+ mice showed a greater tendency for lymph node (35% versus 0%; P = 0.04) and lung (53% versus 22%) metastasis and lower rates of spontaneous apoptosis than those in Survivin−/HGF+ mice. These studies show a role for Survivin in promoting both early and late events of UV-induced melanoma development in vivo. [Cancer Res 2007;67(11):5172–8]
Alternative BCR/ABL Splice Variants in Philadelphia Chromosome–Positive Leukemias Result in Novel Tumor-Specific Fusion Proteins that May Represent Potential Targets for Immunotherapy Approaches
Authors: Volpe Gisella , Cignetti Alessandro , Panuzzo Cristina , Kuka Mirela , Vitaggio Katiuscia , Brancaccio Mara , Perrone Giuseppe , Rinaldi Monica , Prato Giuseppina , Fava Milena , Geuna Massimo , Pautasso Marisa , Casnici Claudia , Signori Emanuela , Tonon Giancarlo , Tarone Guido , Marelli Ornella , Fazio Vito M. , Saglio Giuseppe
Published: June 2007

Imatinib currently represents the standard treatment in the early chronic phase of chronic myelogenous leukemia (CML), thanks to the high percentage of cytogenetic complete remission achieved, but it is yet unclear to what extent it can eradicate leukemia. Therefore, different vaccination strategies have been suggested, mainly based on the exploitment of the junctional peptides spanning the fusion region of the Bcr/Abl proteins. To identify new potential immunologic targets, 63 Philadelphia chromosome–positive patients and 6 BCR/ABL–positive cell lines were tested in nested reverse transcriptase PCR to detect the presence of BCR/ABL transcripts arising from the alternative splicing of the main BCR/ABL transcripts. We could detect BCR/ABL transcripts with junctions between BCR exon 1, 13, or 14 and ABL exon 4 in ∼80% of patients and 84% of cell lines, beside the main fusion transcripts. Translation products of these transcripts were characterized at their COOH terminus by a large amino acid portion derived from the out of frame (OOF) reading of ABL gene. These proteins were detected in BCR/ABL–positive cell lines by immunoprecipitation and immunohistochemistry. Finally, we determined whether OOF-specific CD8+ T cells could be found in the peripheral blood of CML patients and whether they could acquire effector function following in vitro sensitization with OOF-derived peptides predicted to bind to human leucocyte antigen (HLA)-A2 and HLA-A3 molecules. We detected the presence of OOF-specific CD8+ T cells in four of four patients studied, and in one case, these T cells exhibited specific cytotoxic activity against both peptide-pulsed targets and autologous primary CML cells. [Cancer Res 2007;67(11):5300–7]
Protein Kinase Cα Determines HER2 Fate in Breast Carcinoma Cells with HER2 Protein Overexpression without Gene Amplification
Authors: Magnifico Alessandra , Albano Luisa , Campaner Stefano , Campiglio Manuela , Pilotti Silvana , Ménard Sylvie , Tagliabue Elda
Published: June 2007

In some HER2-positive breast tumors, cell surface overexpression of HER2 is not associated with gene amplification but may instead rest in altered gene transcription, half-life, or recycling of the oncoprotein. Here, we show that HER2 overexpression in HER2 2+ carcinomas is associated with neither an increase in gene transcription nor a deregulation in the ubiquitin-dependent pathways, but instead seems to be regulated by protein kinase Cα (PKCα) activity. The stimulation of PKCα up-regulated HER2 expression, whereas PKCα inhibition by pharmacologic treatments and PKCα-specific small interfering RNA led to a dramatic down-regulation of HER2 levels only in breast cancer cells HER2 2+. Consistent with the in vitro data, our biochemical analysis of HER2 2+ human primary breast specimens revealed significantly higher levels of phosphorylated PKCα compared with HER2-negative tumors. Inhibition of HER2 activation by the tyrosine kinase inhibitor lapatinib led to decreased levels of PKCα phosphorylation, clearly indicating a cross-talk between PKCα and HER2 molecules. These data suggest that HER2 overexpression in HER2 2+ carcinomas is due to an accumulation of the recycled oncoprotein to the cell surface induced by activated PKCα. [Cancer Res 2007;67(11):5308–17]
Elevated Interleukin-6 and G-CSF in Human Pancreatic Cancer Cell Conditioned Medium Suppress Dendritic Cell Differentiation and Activation
Authors: Bharadwaj Uddalak , Li Min , Zhang Rongxin , Chen Changyi , Yao Qizhi
Published: June 2007

Although dendritic cell (DC) function is impaired in pancreatic cancer patients, the underlying mechanisms are unknown. This study analyzed the soluble factors released by pancreatic cancer cells responsible for inhibiting DC differentiation and activation. Medium conditioned by a highly metastatic human pancreatic cancer cell line BxPC-3 [BxPC-3 conditioned medium (BxCM)] was mainly used for the study. Both CD34+ hematopoietic progenitor cell–derived and CD14+ monocyte-derived immature DCs and mature DCs (mDCs) were inhibited by BxCM. Allostimulation of CD4+ and CD8+ T cells by BxCM-treated mDCs was inefficient and resulted in production of lower levels of Th1 and Th2 cytokines. Antigen-specific T-cell activation capability was also reduced in BxCM-treated mDCs. Addition of exogenous interleukin-6 (IL-6) and granulocyte colony-stimulating factor (G-CSF), which were present in high amounts in BxCM, mimicked the inhibitory effect of BxCM on DC differentiation and maturation. IL-6 was able to suppress DC differentiation and G-CSF mainly acted on the suppressing allostimulatory capacity of DCs. In addition, pancreatic cancer patient sera were able to inhibit DC differentiation of CD14+ monocytes obtained from healthy donors. Depleting IL-6 or G-CSF from BxCM could reverse the DC-inhibitory properties of BxCM. Furthermore, BxCM, IL-6, or G-CSF led to the activation of signal transducer and activator of transcription 3 (STAT3) in CD14+ monocytes to different degrees. Blocking BxCM-induced STAT3 activation also reversed the inhibitory effect of BxCM on DC differentiation. Therefore, IL-6 and G-CSF in BxCM represent two main factors responsible for suppression of DC differentiation, maturation, and antigen presentation, and this suppression of DC functions may be due to the aberrant activation of STAT3 by BxCM. [Cancer Res 2007;67(11):5479–88]
Histone Deacetylase Inhibitors Sensitize Prostate Cancer Cells to Agents that Produce DNA Double-Strand Breaks by Targeting Ku70 Acetylation
Authors: Chen Chang-Shi , Wang Yu-Chieh , Yang Hsiao-Ching , Huang Po-Hsien , Kulp Samuel K. , Yang Chih-Cheng , Lu Yen-Shen , Matsuyama Shigemi , Chen Ching-Yu , Chen Ching-Shih
Published: June 2007

This study reports a histone deacetylation-independent mechanism whereby histone deacetylase (HDAC) inhibitors sensitize prostate cancer cells to DNA-damaging agents by targeting Ku70 acetylation. Ku70 represents a crucial component of the nonhomologous end joining repair machinery for DNA double-strand breaks (DSB). Our data indicate that pretreatment of prostate cancer cells with HDAC inhibitors (trichostatin A, suberoylanilide hydroxamic acid, MS-275, and OSU-HDAC42) led to increased Ku70 acetylation accompanied by reduced DNA-binding affinity without disrupting the Ku70/Ku80 heterodimer formation. As evidenced by increased Ser139-phosphorylated histone H2AX (γH2AX), impaired Ku70 function diminished cellular capability to repair DNA DSBs induced by bleomycin, doxorubicin, and etoposide, thereby enhancing their cell-killing effect. This sensitizing effect was most prominent when cells were treated with HDAC inhibitors and DNA-damaging agents sequentially. Mimicking acetylation was done by replacing K282, K317, K331, K338, K539, or K542 with glutamine via site-directed mutagenesis, which combined with computer docking analysis was used to analyze the role of these lysine residues in the interactions of Ku70 with DNA broken ends. Mutagenesis of K282, K338, K539, or K542 suppressed the activity of Ku70 to bind DNA, whereas mutagenesis of K317 or K331 with glutamine had no significant effect. Moreover, overexpression of K282Q or K338Q rendered DU-145 cells more susceptible to the effect of DNA-damaging agents on γH2AX formation and cell killing. Overall, the ability of HDAC inhibitors to regulate cellular ability to repair DNA damage by targeting Ku70 acetylation underlies the viability of their combination with DNA-damaging agents as a therapeutic strategy for prostate cancer. [Cancer Res 2007;67(11):5318–27]
T-Lymphokine–Activated Killer Cell–Originated Protein Kinase Functions as a Positive Regulator of c-Jun-NH2-Kinase 1 Signaling and H-Ras–Induced Cell Transformation
Authors: Oh Sang-Muk , Zhu Feng , Cho Yong-Yeon , Lee Ki Won , Kang Bong Seok , Kim Hong-Gyum , Zykova Tatyana , Bode Ann M. , Dong Zigang
Published: June 2007

T-lymphokine–activated killer cell–originated protein kinase (TOPK) is overexpressed in highly proliferating tumors such as leukemias and myelomas, and seems to play a key role in tumorigenesis or metastasis. However, the precise role and regulatory mechanism explaining the effects of TOPK on tumor cells still remain elusive. Here, we reported that TOPK regulates UVB-induced c-Jun-NH2-kinase 1 (JNK1) activity, and is essential for H-Ras–induced activator protein-1 activity and cell transformation. We showed that TOPK associated with and phosphorylated JNK1 following UVB irradiation in vitro or in vivo. Moreover, UVB-induced JNK1 activity was greatly augmented in mouse epidermal JB6 Cl41 cells that stably expressed TOPK cDNA. On the other hand, JNK1 activity was markedly attenuated by stable expression of small interfering RNA against TOPK in malignant melanoma RPMI 7951 cells. Interestingly, TOPK interacted with JNK-interacting protein 1 and caused an elevation of JNK-interacting protein 1 scaffolding activity, thereby enhancing JNK1 activity. Furthermore, JNK1 was required for TOPK-mediated activator protein-1 transcriptional activity and transformed foci induced by UVB or H-Ras. Taken together, these findings showed that TOPK positively modulated UVB-induced JNK1 activity and played a pivotal role in JNK1-mediated cell transformation induced by H-Ras. These studies might also provide a novel molecular mechanism for the role of TOPK in UVB-mediated skin carcinogenesis. [Cancer Res 2007;67(11):5186–94]