Double inhibition of XIAP and Bcl-2 axis is beneficial for retrieving sensitivity of renal cell cancer to apoptosis

Renal cell carcinoma (RCC) is known to be resistant to chemo- and radiotherapy due to a high apoptotic threshold. Smac and XIAP (X-linked inhibitor of apoptosis protein) proteins were detected in all RCC cell lines and tissue samples examined. We modulated the function of XIAP, either through its constitutional downregulation with an shRNA vector or by applying a Smac-mimicking peptide. Among RCC cell lines, Caki1 expresses the highest levels of XIAP. We transfected Caki1 with XIAP-targeting shRNA vector and generated stable clones. XIAP was knocked down by RNA interference in clone no. 14 by 81.6% and in clone no. 19 by 85.3%. Compared to the parental and mock-transfected cells, neither clone was more sensitive to conventional chemotherapeutic agents, but both clones were more susceptible to Fas stimulation (P<0.0001) and to pharmacological Bcl-2 inhibition (P<0.0001), as well as to a combination of the two (P<0.0001). Mature Smac binds to XIAP via the N-terminal residues, disrupting its interaction with caspases and promoting their activity. We determined that exposure of Caki1 cells to Smac-N7 peptide (AVPIAQK) resulted in a slight but significant decrease in viability (P=0.0031) and potentiated cisplatin's effect (P=0.0027). In contrast with point targeting of XIAP by shRNA, Smac-N7 peptide is active against several IAP (inhibitor of apoptosis protein) family members, which can explain its role in sensitising cells to cisplatin. Our results suggest that multiple targeting of both Bcl-2 and XIAP or, alternatively, of several IAP family members by the Smac-N7 peptide is a potent way to overcome resistance of RCC to apoptosis-triggering treatment modalities, and might be a new tool for molecular targeted therapy

Differential regulation of diacylglycerol kinase isoform in human failing hearts

Evidence from several studies indicates the importance of Gαq protein-coupled receptor (GPCR) signaling pathway, which includes diacylglycerol (DAG), and protein kinase C, in the development of heart failure. DAG kinase (DGK) acts as an endogenous regulator of GPCR signaling pathway by catalyzing and regulating DAG. Expressions of DGK isoforms α, ε, and ζ in rodent hearts have been detected; however, the expression and alteration of DGK isoforms in a failing human heart has not yet been examined. In this study, we detected mRNA expressions of DGK isoforms γ, η, ε, and ζ in failing human heart samples obtained from patients undergoing cardiovascular surgery with cardiopulmonary bypass. Furthermore, we investigated modulation of DGK isoform expression in these hearts. We found that expressions of DGKη and DGKζ were increased and decreased, respectively, whereas those of DGKγ and DGKε remained unchanged. This is the first report that describes the differential regulation of DGK isoforms in normal and failing human hearts

The use of a non-standard high calcium fly ash in concrete and its response to accelerated curing [Utilización de una ceniza volante de alto contenido en cal en el hormigón y su comportamiento frente a curados acelerados]

An experimental work was carried out to investigate the use of a non-standard high calcium fly ash in concrete. The response of the same fly ash to the accelerated curing was also explored. With three different cementitious material contents, a total of 48 concretes were produced. The water/cement ratios were varied from 0.40 to 0.87. Compressive strengths of the moist cured cube specimens cast from the concrete mixtures made with 0%, 15%, 30% and 45% replacement of normal Portland cement with fly ash were measured at 28 days and 3 months. Accelerated compressive strengths were also measured using warm-water method and boiling-water method in accordance with the relevant ASTM and Turkish Standards. Despite the fact that the fly ash used was a non-standard, the laboratory test results showed that it could be utilized in concrete production at a replacement level between 15% and 30% by weight basis because fly ash concrete developed comparable or higher compressive strength than that of corresponding normal Portland cement concrete. The laboratory test results also indicated that the accelerated curing could be used to predict the compressive strength of fly ash concrete with 85% correlation coefficient. The amount of fly ash was found to be immaterial in the strength prediction. The relation between warm-water method and boiling-water method was of linear form with 93% correlation coefficient

The use of a non-standard high calcium fly ash in concrete and its response to accelerated curing

WOS: 000178265000002An experimental work was carried out to investigate the use of a non-standard high calcium fly ash in concrete. The response of the same fly ash to the accelerated curing was also explored. With three different cementitious material contents, a total of 48 concretes were produced. The water/cement ratios were varied from 0.40 to 0.87. Compressive strengths of the moist cured cube specimens cast from the concrete mixtures made with 0%, 15%, 30% and 45% replacement of normal Portland cement with fly ash were measured at 28 days and 3 months. Accelerated compressive strengths were also measured using warm water method and boiling-water method in accordance with the relevant ASTM and Turkish Standards. Despite the fact that the fly ash used was a non-standard, the laboratory test results showed that it could be utilized in concrete production at a replacement level between 15% and 30% by weight basis because fly, ash concrete developed comparable or higher compressive strength than that of corresponding normal Portland cement concrete. The laboratory test results also indicated that the accelerated curing could be used to predict the compressive strength of fly ash concrete with 85% correlation coefficient. The amount of fly ash was found to be immaterial in the strength prediction. The relation between warm-water method and boiling-water method was of linear form with 93% correlation coefficient

Influence of dry and wet curing conditions on compressive strength of silica fume concrete

This paper reports a part of an ongoing laboratory investigation in which the compressive strength of silica fume concrete is studied under dry and wet curing conditions. In the study, a total of 48 concretes, including control Portland cement concrete and silica fume concrete, were produced with four different water-cement ratios (0.3, 0.4, 0.5, 0.6), three different cement dosages (350, 400, 450 kg/m3) and three partial silica fume replacement ratios (10%, 15%, 20%). A hyperplastisizer was used in concrete at various quantities to provide and keep a constant workability. Three cubic samples produced from fresh concrete were demoulded after a day; then, they were cured at 20±2 °C with 65% relative humidity (RH), and three other cubic samples were cured at 20±2 °C with 100% RH until the samples were used for compressive strength measurement at 28 days. The comparison was made on the basis of compressive strength between silica fume concrete and control Portland cement concrete. Silica fume concretes were also compared among themselves. The comparisons showed that compressive strength of silica fume concrete cured at 65% RH was influenced more than that of Portland cement concrete. It was found that the compressive strength of silica fume concrete cured at 65% RH was, at average, 13% lower than that of silica fume concrete cured at 100% RH. The increase in the water-cementitious material ratios makes the concrete more sensitive to dry curing conditions. The influence of dry curing conditions on silica fume concrete was marked as the replacement ratio of silica fume increased. © 2005 Elsevier Ltd. All rights reserved

Influence of dry and wet curing conditions on compressive strength of silica fume concrete

This paper reports a part of an ongoing laboratory investigation in which the compressive strength of silica fume concrete is studied under dry and wet curing conditions. In the study, a total of 48 concretes, including control Portland cement concrete and silica fume concrete, were produced with four different water-cement ratios (0.3, 0.4, 0.5, 0.6), three different cement dosages (350, 400, 450kg/m(3)) and three partial silica fume replacement ratios (10%, 15%, 20%). A hyperplastisizer was used in concrete at various quantities to provide and keep a constant workability. Three cubic samples produced from fresh concrete were demoulded after a day; then, they were cured at 20 +/- 2 degrees C with 65% relative humidity (RH), and three other cubic samples were cured at 20 +/- 2 degrees C with 100% RH until the samples were used for compressive strength measurement at 28 days. The comparison was made on the basis of compressive strength between silica fume concrete and control Portland cement concrete. Silica fume concretes were also compared among themselves. The comparisons showed that compressive strength of silica fume concrete cured at 65% RH was influenced more than that of Portland cement concrete. It was found that the compressive strength of silica fume concrete cured at 65% RH was, at average, 13% lower than that of silica fume concrete cured at 100% RH. The increase in the water-cementitious material ratios makes the concrete more sensitive to dry curing conditions. The influence of dry curing conditions on silica fume concrete was marked as the replacement ratio of silica fume increased. (c) 2005 Elsevier Ltd. All rights reserved