Mechanical Properties of Polyvinyl Alcohol Fiber-Reinforced Cementitious Composites after High-Temperature Exposure.

The mechanical properties of cementitious composites before and after exposure to high temperature are affected by calcium-silicate-hydrate (C-S-H) gels. To evaluate the effects of high temperature, plyvinyl alcohol (PVA) fiber content, and the cooling method on properties of cementitious composites, physical, mechanical, and microscopic tests were performed in this study. The target temperatures were 25, 100, 200, 300, 400, 600, and 800 °C. The PVA fiber contents were 0.0, 0.3, 0.6, 0.9, 1.2, and 1.5 vol%. The high-temperature resistance of PVA fiber-reinforced cementitious composite (PVA-FRCC) specimens was investigated through changes in their appearance, mass loss, compressive strength, splitting tensile strength, flexural strength, and microstructure. The results showed that PVA fibers reduced the probability of explosion spalling in the PVA-FRCC specimens exposed to high temperatures. The mass loss rate of samples exposed to temperatures below 200 °C was small and lower than 5%, whereas a significant mass loss was observed at 200 °C to 800 °C. A small rise in the cubic compressive and splitting tensile strengths of samples was found at 400 °C and 300 °C, respectively. Below 400 °C, the fibers were beneficial to the mechanical strength of the PVA-FRCC specimens. Nevertheless, when the temperature was heated above 400 °C, melted fibers created many pores and channels, which caused a decrease in the strength of the specimens. The method of cooling with water could aggravate the damage to the cementitious composites exposed to temperatures above 200 °C. High temperature could lead to the decomposition of the C-S-H gels of the PVA-FRCC samples, which makes C-S-H gels lose their bonding ability. From the perspective of the microstructure, the structure of PVA-FRCC samples exposed to 600 °C and 800 °C became loose and the number of microcracks increased, which confirmed the reduction in macro-mechanical properties.

The Evaluation of Construction Dust Diffusion and Sedimentation Using Wind Tunnel Experiment.

A large quantity of particulate matter is generated during construction of civil engineering projects, which has a negative effect on the atmosphere and environment. In order to explore the concentration, distribution and diffusion of particulate matters generated from construction dust with different moisture contents, a wind tunnel experiment was conducted, and the effects of wind speed and moisture content on the inhibition rate, drifting distance and suppression percentage of particulate matters were investigated. The results show that the peak concentration decreases with the increase in moisture content, compared with dry dust; the peak concentrations for 1%, 2% and 3% moisture content are reduced by 37.07%, 39.53% and 65.38%, respectively. The average concentrations in the cross-section decrease with the increase in the moisture content, resulting in an increasing tendency of the particle inhibition rate. The forecast drifting distance decreases with the increase in the moisture content; when the suspension percentage is 1%, the forecast drifting distances of dry dust, 1%, 2% and 3% moisture content are 641.58, 116.08, 19.33 and 3.82 km, respectively, for a 5 m/s wind speed. Considering that an increase in wind velocity will not only decrease the inhibition rate but also increase the drifting distance, the dust suppression method by increasing the moisture content in low and medium wind velocities is applicable. When the limit value of the particle suppression rate within a distance of 50 m is larger than 70%, construction activities are prohibited at any wind velocity for dry and 1% moisture content, and at wind velocities larger than 2 m/s and 4 m/s for 2% and 3% moisture content, respectively.

Mechanical Properties of Nano-SiO2 Reinforced Geopolymer Concrete under the Coupling Effect of a Wet-Thermal and Chloride Salt Environment.

In this study, the mechanical behaviors of nano-SiO2 reinforced geopolymer concrete (NS-GPC) under the coupling effect of a wet-thermal and chloride salt environment were investigated through a series of basic experiments, and a simulation on the coupling effect of a wet-thermal and chloride salt environment and SEM test were also included. During the experiments for the coupling effect of the wet-thermal and chloride salt environment, an environment simulation test chamber was utilized to simulate the wet-thermal and chloride salt environment, in which the parameters of relative humidity, temperature, mass fraction of NaCl solution and action time were set as 100%, 45 °C, 5% and 60 d, respectively. The content of nano-SiO2 (NS) particles added in geopolymer concrete (GPC) were 0, 0.5%, 1.0%, 1.5% and 2.0%. The result indicated that the mechanical properties of NS reinforced GPC decreased under the coupling effect of the wet-thermal and chloride salt environment compared to the control group in the natural environment. When the NS content was 1.5%, the cube and splitting tensile strength, elastic modulus and impact toughness of GPC under the coupling environment of wet-thermal and chloride salt were decreased by 9.7%, 9.8%, 19.2% and 44.4%, respectively, relative to that of the GPC under the natural environment. The addition of NS improved the mechanical properties of GPC under the coupling effect of the wet-thermal and chloride salt environment. Compared to the control group without NS, the maximum increment in cube compressive strength, splitting tensile strength and elastic modulus of NS-GPC under the coupling effect of the wet-thermal and chloride salt environment due to the incorporation of NS reached 25.8%, 9.6% and 17.2%, respectively. Specifically, 1.5% content of NS increased the impact toughness, impact numbers of initial crack and the ultimate failure of GPC by 122.3%, 109% and 109.5%, respectively.

Early-Age Mechanical Characteristics and Microstructure of Concrete Containing Mineral Admixtures under the Environment of Low Humidity and Large Temperature Variation.

The application of concrete containing mineral admixtures was attempted in Northwest China in this study, where the environment has the characteristics of low humidity and large temperature variation. The harsh environment was simulated by using an environmental chamber in the laboratory and four types of concrete were prepared, including ordinary concrete and three kinds of mineral admixture concretes with different contents of fly ash and blast-furnace slag. These concretes were cured in the environmental chamber according to the real curing conditions during construction. The compression strength, fracture properties, SEM images, air-void characteristics, and X-ray diffraction features were researched at the early ages of curing before 28 d. The results showed that the addition of fly ash and slag can improve the compression strength and fracture properties of concrete in the environment of low humidity and large temperature variation. The optimal mixing of mineral admixture was 10% fly ash and 20% slag by replacing the cement in concrete, which can improve the compression strength, initial fracture toughness, unstable fracture toughness, and fracture energy by 23.9%, 25.2%, 45.3%, and 22.6%, respectively, compared to ordinary concrete. Through the analysis of the microstructure of concrete, the addition of fly ash and slag can weaken the negative effects of the harsh environment of low humidity and large temperature variation on concrete microstructure and cement hydration.

Protein kinase CK2 inhibition induces cell death via early impact on mitochondrial function.

CK2 (official acronym for casein kinase 2 or II) is a potent suppressor of apoptosis in response to diverse apoptotic stimuli-thus its molecular downregulation or activity inhibition results in potent induction of cell death. CK2 downregulation is known to impact mitochondrial apoptotic circuitry but the underlying mechanism(s) remain unclear. Utilizing prostate cancer cell lines subjected to CK2-specific inhibitors which cause loss of cell viability, we have found that CK2 inhibition in cells causes rapid early decrease in mitochondrial membrane potential (Δψm). Cells treated with the CK2 inhibitors TBB (4,5,6,7-tetrabromobenzotriazole) or TBCA (tetrabromocinnamic acid) demonstrate changes in Δψm which become apparent within 2 h, that is, significantly prior to evidence of activation of other mitochondrial apoptotic signals whose temporal expression ensues subsequent to loss of Δψm. Further, we have demonstrated the presence of CK2 in purified mitochondria and it appears that the effect on Δψm evoked by inhibition of CK2 may involve mitochondrial localized CK2. Results also suggest that alterations in Ca(2+) signaling may be involved in the CK2 mediated regulation of Δψm and mitochondrial permeability. Thus, we propose that a key mechanism of CK2 impact on mitochondrial apoptotic circuitry and cell death involves early loss of Δψm which may be a primary trigger for apoptotic signaling and cell death resulting from CK2 inhibition.

The Notch pathway is important in maintaining the cancer stem cell population in pancreatic cancer.

BACKGROUND: Pancreatic cancer stem cells (CSCs) represent a small subpopulation of pancreatic cancer cells that have the capacity to initiate and propagate tumor formation. However, the mechanisms by which pancreatic CSCs are maintained are not well understood or characterized. METHODS: Expression of Notch receptors, ligands, and Notch signaling target genes was quantitated in the CSC and non-CSC populations from 8 primary human pancreatic xenografts. A gamma secretase inhibitor (GSI) that inhibits the Notch pathway and a shRNA targeting the Notch target gene Hes1 were used to assess the role of the Notch pathway in CSC population maintenance and pancreatic tumor growth. RESULTS: Notch pathway components were found to be upregulated in pancreatic CSCs. Inhibition of the Notch pathway using either a gamma secretase inhibitor or Hes1 shRNA in pancreatic cancer cells reduced the percentage of CSCs and tumorsphere formation. Conversely, activation of the Notch pathway with an exogenous Notch peptide ligand increased the percentage of CSCs as well as tumorsphere formation. In vivo treatment of orthotopic pancreatic tumors in NOD/SCID mice with GSI blocked tumor growth and reduced the CSC population. CONCLUSION: The Notch signaling pathway is important in maintaining the pancreatic CSC population and is a potential therapeutic target in pancreatic cancer.

Protein kinase A modulates transforming growth factor-ß signaling through a direct interaction with Smad4 protein.

Transforming growth factor ß (TGFß) signaling normally functions to regulate embryonic development and cellular homeostasis. It is increasingly recognized that TGFß signaling is regulated by cross-talk with other signaling pathways. We previously reported that TGFß activates protein kinase A (PKA) independent of cAMP through an interaction of an activated Smad3-Smad4 complex and the regulatory subunit of the PKA holoenzyme (PKA-R). Here we define the interaction domains of Smad4 and PKA-R and the functional consequences of this interaction. Using a series of Smad4 and PKA-R truncation mutants, we identified amino acids 290-300 of the Smad4 linker region as critical for the specific interaction of Smad4 and PKA-R. Co-immunoprecipitation assays showed that the B cAMP binding domain of PKA-R was sufficient for interaction with Smad4. Targeting of B domain regions conserved among all PKA-R isoforms and exposed on the molecular surface demonstrated that amino acids 281-285 and 320-329 were required for complex formation with Smad4. Interactions of these specific regions of Smad4 and PKA-R were necessary for TGFß-mediated increases in PKA activity, CREB (cAMP-response element-binding protein) phosphorylation, induction of p21, and growth inhibition. Moreover, this Smad4-PKA interaction was required for TGFß-induced epithelial mesenchymal transition, invasion of pancreatic tumor cells, and regulation of tumor growth in vivo.

Anti-DLL4 has broad spectrum activity in pancreatic cancer dependent on targeting DLL4-Notch signaling in both tumor and vasculature cells.

PURPOSE: We previously showed that targeting Delta-like ligand 4 (DLL4) in colon and breast tumors inhibited tumor growth and reduced tumor initiating cell frequency. In this report, we have extended these studies to pancreatic cancer and probed the mechanism of action in tumor and stromal cells involved in antitumor efficacy. EXPERIMENTAL DESIGN: Patient-derived pancreatic xenograft tumor models were used to evaluate the antitumor effect of anti-DLL4. To investigate the mechanism of action, we compared the activity of targeting DLL4 in tumor cells with an anti-human DLL4 antibody (anti-hDLL4) and in the host stroma/vasculature with an anti-mouse DLL4 antibody (anti-mDLL4). The effect of these antibodies on cancer stem cell frequency was examined by in vivo limiting dilution assays. RESULTS: The combination of anti-hDLL4 and anti-mDLL4 was efficacious in a broad spectrum of pancreatic tumor xenografts and showed additive antitumor activity together with gemcitabine. Treatment with either anti-hDLL4 or anti-mDLL4 delayed pancreatic tumor recurrence following termination of gemcitabine treatment, and the two together produced an additive effect. Anti-hDLL4 had a pronounced effect in reducing the tumorigenicity of pancreatic cancer cells based on serial transplantation and tumorsphere assays. In contrast, disruption of tumor angiogenesis with anti-mDLL4 alone or with anti-VEGF had minimal effects on tumorigenicity. Gene expression analyses indicated that anti-DLL4 treatment regulated genes that participate in Notch signaling, pancreatic differentiation, and epithelial-to-mesenchymal transition. CONCLUSIONS: Our findings suggest a novel therapeutic approach for pancreatic cancer treatment through antagonism of DLL4/Notch signaling.

Hypoxic-ischemic injury decreases anxiety-like behavior in rats when associated with loss of tyrosine-hydroxylase immunoreactive neurons of the substantia nigra

Neonatal Sprague-Dawley rats were randomly divided into normal control, mild hypoxia-ischemia (HI), and severe HI groups (N = 10 in each group at each time) on postnatal day 7 (P7) to study the effect of mild and severe HI on anxiety-like behavior and the expression of tyrosine hydroxylase (TH) in the substantia nigra (SN). The mild and severe HI groups were exposed to hypoxia (8 percent O2/92 percent N2) for 90 and 150 min, respectively. The elevated plus-maze (EPM) test was performed to assess anxiety-like behavior by measuring time spent in the open arms (OAT) and OAT percent, and immunohistochemistry was used to determine the expression of TH in the SN at P14, P21, and P28. OAT and OAT percent in the EPM were significantly increased in both the mild (1.88-, 1.99-, and 2.04-fold, and 1.94-, 1.51-, and 1.46-fold) and severe HI groups (1.69-, 1.68-, and 1.87-fold, and 1.83-, 1.43-, and 1.39-fold, respectively; P < 0.05). The percent of TH-positive cells occupying the SN area was significantly and similarly decreased in both the mild (17.7, 40.2, and 47.2 percent) and severe HI groups (16.3, 32.2, and 43.8 percent, respectively; P < 0.05). The decrease in the number of TH-positive cells in the SN and the level of protein expression were closely associated (Pearson correlation analysis: r = 0.991, P = 0.000 in the mild HI group and r = 0.974, P = 0.000 in the severe HI group) with the impaired anxiety-like behaviors. We conclude that neonatal HI results in decreased anxiety-like behavior during the juvenile period of Sprague-Dawley rats, which is associated with the decreased activity of TH in the SN. The impairment of anxiety and the expression of TH are not likely to be dependent on the severity of HI.

Hypoxic-ischemic injury decreases anxiety-like behavior in rats when associated with loss of tyrosine-hydroxylase immunoreactive neurons of the substantia nigra.

Neonatal Sprague-Dawley rats were randomly divided into normal control, mild hypoxia-ischemia (HI), and severe HI groups (N = 10 in each group at each time) on postnatal day 7 (P7) to study the effect of mild and severe HI on anxiety-like behavior and the expression of tyrosine hydroxylase (TH) in the substantia nigra (SN). The mild and severe HI groups were exposed to hypoxia (8% O2/92% N2) for 90 and 150 min, respectively. The elevated plus-maze (EPM) test was performed to assess anxiety-like behavior by measuring time spent in the open arms (OAT) and OAT%, and immunohistochemistry was used to determine the expression of TH in the SN at P14, P21, and P28. OAT and OAT% in the EPM were significantly increased in both the mild (1.88-, 1.99-, and 2.04-fold, and 1.94-, 1.51-, and 1.46-fold) and severe HI groups (1.69-, 1.68-, and 1.87-fold, and 1.83-, 1.43-, and 1.39-fold, respectively; P < 0.05). The percent of TH-positive cells occupying the SN area was significantly and similarly decreased in both the mild (17.7, 40.2, and 47.2%) and severe HI groups (16.3, 32.2, and 43.8%, respectively; P < 0.05). The decrease in the number of TH-positive cells in the SN and the level of protein expression were closely associated (Pearson correlation analysis: r = 0.991, P = 0.000 in the mild HI group and r = 0.974, P = 0.000 in the severe HI group) with the impaired anxiety-like behaviors. We conclude that neonatal HI results in decreased anxiety-like behavior during the juvenile period of Sprague-Dawley rats, which is associated with the decreased activity of TH in the SN. The impairment of anxiety and the expression of TH are not likely to be dependent on the severity of HI.

c-Met is a marker of pancreatic cancer stem cells and therapeutic target.

BACKGROUND & AIMS: Growth of many different tumor types requires a population of self-renewing cancer stem cells (CSCs). c-Met is a marker of normal mouse pancreatic stem and progenitor cells; we investigated whether it is also a marker of human pancreatic CSCs that might be developed as a therapeutic target. METHODS: We studied growth of primary human pancreatic adenocarcinoma in NOD SCID mice. The self-renewal capability of pancreatic cancer cells that expressed high levels of c-Met (c-Met(high)) was assessed using in vitro sphere assays and compared with those that were c-Met negative or expressed low levels of c-Met. The tumorigenicity of c-Met(high) pancreatic cancer cells was evaluated in NOD SCID mice. RESULTS: c-Met(high) cells readily formed spheres, whereas c-Met-negative cells did not. Use of the c-Met inhibitor XL184 or c-Met knockdown with small hairpin RNAs significantly inhibited tumor sphere formation. c-Met(high) cells had increased tumorigenic potential in mice; those that expressed c-Met and CD44 (0.5%-5% of the pancreatic cancer cells) had the capability for self-renewal and the highest tumorigenic potential of all cell populations studied. In pancreatic tumors established in NOD SCID mice, c-Met inhibitors slowed tumor growth and reduced the population of CSCs when given alone or in combination with gemcitabine. Administration of XL184 for 2 weeks after cardiac injection of cancer cells prevented the development of metastases. CONCLUSIONS: c-Met is a new marker for pancreatic CSCs. It is required for growth and metastasis of pancreatic tumors in mice and is a therapeutic target for pancreatic cancer.

Characterization of a core binding site for ADAMTS-13 in the A2 domain of von Willebrand factor.

ADAMTS-13, a metalloprotease in plasma, specifically cleaves the Tyr-1605-Met-1606 bond in the A2 domain of von Willebrand factor (VWF) to regulate the polymer distribution of VWF in circulation, which is critical for primary hemostasis. A 73-aa peptide (VWF73) was previously identified as the minimal substrate cleavable by ADAMTS-13. In this study, VWF73 was enzymatically and chemically cleaved into shorter peptides, and the inhibition of cleavage of a VWF73-derived substrate by these purified peptides was measured in competition studies using a quantitative assay we recently reported. A 24-aa peptide encompassing Pro-1645-Lys-1668 (P'40-P'63) and situated 40 aa downstream from the cleavage site was the minimal peptide that could bind to and competitively inhibit ADAMTS-13 (K(i) = 12 microM). This peptide and longer peptides encompassing this core sequence also inhibited the cleavage of multimeric VWF by ADAMTS-13. These results suggest the presence of a complementary extended binding site, or exosite, on ADAMTS-13. Mutation of Asp-1653 and Asp-1663 to Ala in this region significantly reduced the rate of cleavage of the substrate peptide, whereas the Glu1655Ala mutation caused an enhanced rate of cleavage. These results suggest that ionic interactions of the Pro-1645-Lys-1668 region with the exosite on ADAMTS-13 play a significant role in mediating substrate recognition.

Brownian dynamics simulations of the recognition of the scorpion toxin P05 with the small-conductance calcium-activated potassium channels.

The recognition of the scorpion toxin P05 and the small-conductance, calcium-activated potassium (SK) channels, rsk1, rsk2, and rsk3, has been studied by means of the Brownian dynamics (BD) method. All of the 25 available structures of P05 in the RCSB Protein Data Bank determined by NMR were considered during the simulation, which indicated that the conformation of P05 affects both the recognition and binding between the two proteins significantly. Comparing the top four high-frequency structures of P05 binding to the SK channels, we found that the rsk2 channel, with high frequencies and lowest electrostatic interaction energies (E (int)(ES)), is the most favorable for P05 binding, while rsk3 is intermediate, and rsk1 is the least favorable. Among the 25 structures of P05, the 13th structure docks into the binding site of the rsk2 channel with the highest probability and most favorable electrostatic interactions. From the P05-rsk2 channel binding model, we identified the residues critical for the recognition of these two proteins through triplet contact analyses. P05 locates around the extracellular mouth of the SK channels and contacts the SK channels using its alpha-helix rather than beta-sheets. The critical triplet contacts for recognition between P05 and the rsk2 channel are Arg6 (P05)-Asp364 (SK), Arg7 (P05)-Asn368 (SK), and Arg13 (P05)-Asp341 (SK). The structure of the P05-rsk2 complex with the most favorable electrostatic interaction energy was further refined by molecular mechanics, showing that six hydrogen bonding interactions exist between P05 and the rsk2 channel: one hydrogen bond is formed between Arg6 (P05) and Asp364(D) (rsk2); Arg7 (P05) forms three hydrogen bonds with Asp341(B) (rsk2)) and Asp364(C) (rsk2); two hydrogen bonds are formed by Arg13 (P05) with Asp341(A) (rsk2) and Asp364(B) (rsk2). The simulation results are in good agreement with the previous molecular biological experiments and can explain the binding phenomena between P05 and SK channels at the level of molecular structure. The consistency between the results of the BD simulations and the experimental data indicated that our 3D model of the P05-rsk2 channel complex is reasonable and can be employed in further biological studies, such as rational design of the novel therapeutic agents blocking the small-conductance, calcium-activated and apamin-sensitive potassium channels, and for mutagenesis studies in both toxins and SK channels. In particular, both the BD simulations and the molecular mechanics refinements indicate that residue Asp364 of the rsk2 channel is critical for its recognition and binding functionality towards P05. This phenomenon has not been appreciated in the previous mutagenesis experiments, indicating that this might be a new clue for further functional study of SK channels.

Gene expression, mutation, and structure-function relationship of scorpion toxin BmP05 active on SK(Ca) channels.

Four peptide inhibitors of small-conductance Ca(2+)-activated, apamin-sensitive K(+) channels (SK(Ca)) have been isolated from the venom of the Chinese scorpion Buthus martensi, named BmP01, BmP02, BmP03, and BmP05, respectively [Romi-Lebrun, R. (1997) Eur. J. Biochem. 245, 457-464]. Among them BmP05 with 31 amino acid residues has been intensively studied due to its most potent toxicity. To investigate the structure-function relationship of BmP05, its wild type and seven mutants (their C-termini unamidated) were successfully expressed in the yeast secretion system and purified with a high yield over 8 mg/L. Their toxicity to mice and electrophysiological activity on the K(+) currents (SK(Ca) and Kv) in rat adrenal chromaffin cells were measured and compared. The results indicated the following: (1) As a selective antagonist against SK(Ca), 1 microM rBmP05 is equivalent to 0.2 microM apamin, and its IC(50) is 0.92 microM. (2) The basic residues Lys and Arg located at positions 6 and 13 in the N-terminal alpha-helix region are essential and synergetic in the interaction of the toxin with SK(Ca). (3) Disruption of the alpha-helix by mutation of Gln at position 9 with Pro results in almost total loss of toxicity. (4) The C-terminal residue His31 plays an auxiliary role in the interaction of the toxin with SK(Ca). (5) The beta-turn connecting two beta-sheets near the C-terminal part is responsible for the specificity of the toxin to the different subtypes of K(+) channels.