Artificial lightweight aggregates as utilization for future ashes - A case study.

In the future, more electricity in the Netherlands will be produced using coal with co-combustion. Due to this, the generated annual ash volume will increase and the chemical composition will be influenced. One of the options for utilization if present markets are saturated and for use of fly ashes with different compositions, is as raw material for lightweight aggregates. This was selected as one of the best utilizations options regarding potential ash volume to be applied, environmental aspects and status of technology. Because of this, a study has been performed to assess the potential utilization of fly ash for the production of lightweight aggregate. Lightweight aggregate has been produced in a laboratory scale rotary kiln. The raw material consisted of class F fly ash with high free lime content. An addition of 8% clay was necessary to get green pellets with sufficient green strength. The basic properties of the produced lightweight aggregate and its behaviour in concrete have been investigated. The concrete has a good compressive strength and its leaching behaviour meets the most stringent requirements of Dutch environmental regulations. The carbon foot print of concrete will be negatively influenced if only the concrete itself is taken into account, but the reduction of the volume weight has advantages regarding design, transport emissions and isolation properties which may counteract this. In the Dutch situation the operational costs are higher than expected potential selling price for the LWA, which implies that the gate fee for the fly ash is negative.

Recombinant factor VIIa to correct coagulopathy in patients with traumatic brain injury presenting to outlying facilities before transfer to the regional trauma center.

Timely correction of coagulopathy in patients with traumatic brain injury (TBI) improves mortality. Recombinant, activated factor VII (VIIa) has been identified as an effective method to correct coagulopathy in patients with TBI. We performed a retrospective study (January 1, 2008-December 31, 2009) of all patients with TBI and coagulopathy (international normalized ratio (INR) > 1.5) transferred to our Level I trauma center. Twenty-three patients with coagulopathy and TBI were transferred to our trauma center, 100 per cent sustained a fall, and 100 per cent were taking warfarin at the time of injury. Ten patients received VIIa to correct coagulopathy before transfer, whereas 13 did not. The purpose of this study was to compare outcomes in patients who received VIIa with those who did not. When comparing the VIIa group with the no-VIIa group there was no difference in age, gender, Glasgow Coma Scale score, injury severity score, transfer time, or INR at outlying facility. Both groups received one unit of plasma before arrival at our trauma center; patients in the VIIa group received a single 1.2 mg dose of VIIa at the outlying facility. Upon arrival to our trauma center the VIIa group had a lower INR (1.0 vs 3.0, P = 0.02) and lower mortality (0% vs 39%, P = 0.03). In coagulopathic patients with TBI presenting to outlying institutions with limited resources to quickly provide plasma, VIIa efficiently corrects coagulopathy before transfer to definitive care at the regional trauma center. More rapid correction of coagulopathy with VIIa in this patient population may improve mortality.

K-Ras mutant fraction in A/J mouse lung increases as a function of benzo[a]pyrene dose.

K-Ras mutant fraction (MF) was measured to examine the default assumption of low-dose linearity in the benzo[a]pyrene (B[a]P) mutational response. Groups of 10 male A/J mice (7- to 9-weeks old) received a single i.p. injection of 0, 0.05, 0.5, 5, or 50 mg/kg B[a]P and were sacrificed 28 days after treatment. K-Ras codon 12 TGT and GAT MFs in lung DNAs were measured using Allele-specific Competitive Blocker-PCR (ACB-PCR). The K-Ras codon 12 TGT geometric mean MF was 3.88 x 10(-4) in controls, indicating an average of 1 mutation in every approximately 1,288 lung cells. The K-Ras codon 12 TGT geometric mean MFs were as follows: 3.56 x 10(-4); 6.19 x 10(-4); 2.02 x 10(-3), and 3.50 x 10(-3) for the 0.05, 0.5, 5, and 50 mg/kg B[a]P treatment groups, respectively. The 5 and 50 mg/kg dose groups had TGT MFs significantly higher than did controls. Although 10(-5) is considered as the limit of accurate ACB-PCR quantitation, K-Ras codon 12 GAT geometric mean MFs were as follows: 8.38 x 10(-7), 1.47 x 10(-6), 2.19 x 10(-6), 5.71 x 10(-6), and 8.99 x 10(-6) for the 0, 0.05, 0.5, 5, and 50 mg/kg B[a]P treatment groups, respectively. The K-Ras TGT and GAT MFs increased in a B[a]P-dose-dependent manner, with response approximately linear over the 0.05 to 5 mg/kg dose range. K-Ras MF increased with B[a]P adduct burden measured for identical doses in a separate study. Thus, ACB-PCR may be useful in characterizing the shape of a dose-response curve at low doses and establishing relationships between DNA adducts and tumor-associated mutations.

Proteins associated with Culex nigripalpus nucleopolyhedrovirus occluded virions.

Occlusion-derived virions (ODVs) of the nucleopolyhedrovirus of Culex nigripalpus (CuniNPV) were purified by Ludox density gradient ultracentrifugation, and the proteins were separated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Proteins were identified by using Edman sequencing, matrix-assisted laser desorption ionization-time of flight mass spectrometry, nanoelectrospray quadrupole time-of-flight mass spectrometry, or a combination of these methods. Half of the 44 polypeptide sequences identified in this analysis were unique open reading frames (ORFs) encoded by the CuniNPV genome and did not show similarity to any other sequences present in protein databases. Of the 22 polypeptides that showed similarities to other baculovirus-encoded proteins, only 17 sequences have previously been identified as structural proteins. The newly identified CuniNPV structural proteins cun058, cun059, cun087, cun106, and cun109 are homologues of Autographa californica nucleopolyhedrovirus (AcMNPV) ORFs 68, 62, 98, 81, and 2, respectively. The products of four genes, namely, lef-1 (cun045), alkaline exonuclease (cun054), helicase (cun089), and DNA polymerase (cun091), were not detected in the CuniNPV ODV preparations. These four genes are conserved among all annotated baculovirus genomes, and their homologues have been detected in the ODV of AcMNPV.

Biological and molecular studies of a cypovirus from the black fly Simulium ubiquitum (Diptera: Simuliidae).

A cypovirus from the black fly Simulium ubiquitum (SuCPV) was isolated and examined using biological and molecular techniques. SuCPV produces small (typically 0.25mum), polyhedral shaped inclusion bodies (polyhedra), in which the virus particles become multiply embedded. SuCPV is the third cypovirus isolated from Diptera, but the first from Simuliidae that has been characterized using molecular analyses. SuCPV has a genome composed of 10 segments of dsRNA, with an electrophoretic migration pattern that is different from those of recent UsCPV-17 and CrCPV-17 isolates from the mosquitoes Uranotaenia sapphirina and Culex restuans, respectively. The SuCPV electropherotype appears to show significant differences from those of the previously characterized lepidopteran cypoviruses. Sequence analysis of SuCPV segment 10 shows that it is unrelated to either of the two CPV isolates from Diptera or to the CPV species for which Seg-10 has been previously characterized from Lepidoptera. A comparison of the terminal regions of SuCPV genome segments to those of CPV-1, 2, 4, 5 14, 15, 16, 17, 18, and 19 also revealed only low levels of conservation. We therefore, propose that SuCPV is classified within a new Cypovirus species, which we have tentatively identified as Cypovirus-20. We have therefore referred to this virus isolate as S. ubiquitum CPV-20 (SuCPV-20).

Characterizing the residue level folding of the intrinsically unstructured IA3.

Residue level analysis of the folding of simple proteins may hold the key to understanding folding pathways and aid in structure prediction. IA(3), the endogenous inhibitor of yeast aspartic proteinase A (YPrA), is an unstructured protein in solution. Comparison of the 2D (15)N-HSQC spectra of IA(3) in water and in 23% 2,2,2-trifluoroethanol (TFE) shows that the individual residue cross peaks of IA(3) become more dispersed in the presence of TFE, indicating that the protein undergoes an unstructured to structured transition in the presence of TFE. This transition can be monitored by the movements of the cross peaks. Following the individual cross peaks, however, is complicated and does not establish whether a single transition occurs globally in the sequence. In this equilibrium study, we apply singular value decomposition (SVD) to elucidate both the main features of the TFE-driven transition and the residue-level deviations from the average behavior. This analysis has yielded a two-state folding description as well as specifics of NMR frequency shifts of individual residues, indicating that the N-terminus of IA(3) has a higher helical propensity than the C-terminus. Additionally, we discuss possible mechanisms for observed deviations from a two-state folding transition. When combined with a traditional biochemical understanding of interactions between individual residues, this approach leads to a better understanding of protein folding.

Molecular analysis of an occlusion body protein from Culex nigripalpus nucleopolyhedrovirus (CuniNPV).

CUN085 is an occlusion body (OB) protein from the nucleopolyhedrovirus of Culex nigripalpus (CuniNPV). SDS-PAGE analysis indicated that the CuniNPV OB protein is about 3 times the size (approximately 90 kDa) of characterized nucleopolyhedrovirus (NPVs) and granulovirus OB proteins. Rapid amplification of cDNA ends (RACE), RNase protection assay, real-time PCR, and protein sequencing were used to characterize CUN085 from CuniNPV. RACE data revealed that the transcriptional start and termination sites for the CUN085 gene yielded a polypeptide comprised of 822 amino acids indicating that translation initiates within a larger 882 amino acid open reading frame that was originally predicted from the CuniNPV genome sequence. Transcription of CUN085 started at a consensus baculovirus late transcription start site TAAG at nucleotide position 75433 of the CuniNPV genome sequence. RNase protection assays and quantitative real-time PCR show that the CUN085 transcript is first detected in mosquito larvae at approximately 6 h after infection with CuniNPV and its prevalence increased progressively over the subsequent 18 h.

Molecular and biological characterization of a Cypovirus from the mosquito Culex restuans.

A cypovirus from the mosquito Culex restuans (named CrCPV) was isolated and its biology, morphology, and molecular characteristics were investigated. CrCPV is characterized by small (0.1-1.0 microm), irregularly shaped inclusion bodies that are multiply embedded. Laboratory studies demonstrated that divalent cations influenced transmission of CrCPV to Culex quinquefasciatus larvae; magnesium enhanced CrCPV transmission by approximately 30% while calcium inhibited transmission. CrCPV is the second cypovirus from a mosquito that has been confirmed by using molecular analysis. CrCPV has a genome composed of 10 dsRNA segments with an electropherotype similar to the recently discovered UsCPV-17 from the mosquito Uranotaenia sapphirina, but distinct from the lepidopteran cypoviruses BmCPV-1 (Bombyx mori) and TnCPV-15 (Trichoplusia ni). Nucleotide and deduced amino acid sequence analysis of CrCPV segment 10 (polyhedrin) suggests that CrCPV is closely related (83% nucleotide sequence identity and 87% amino acid sequence identity) to the newly characterized UsCPV-17 but is unrelated to the 16 remaining CPV species from lepidopteran hosts. A comparison of the terminal segment regions of CrCPV and UsCPV-17, an additional method for differentiating various Cypovirus species, revealed a high level of conservation. Therefore, we propose that CrCPV is a member of the Cypovirus-17 group and designate this species as CrCPV-17.

Towards an understanding of the oxidation state of cobalt and manganese ions in framework substituted microporous aluminophosphate redox catalysts: An electron paramagnetic resonance and X-ray absorption spectroscopy investigation.

The local structure of manganese and cobalt ions in both the AlPO-5 and AlPO-18 materials was determined using electron paramagnetic resonance (EPR) and X-ray absorption spectroscopy (XAS). This study strongly suggested that both cobalt and manganese ions are present in the framework tetrahedral sites in the as-prepared materials. Upon calcination, in a dry atmosphere, both manganese and cobalt ions are partially oxidised to the 3+ state; the fraction of oxidised cobalt or manganese ions depends upon both the nature of the framework structure and on the ion type with manganese systems showing a higher proportion of oxidised cations.

Morphological and molecular characterization of a Cypovirus (Reoviridae) from the mosquito Uranotaenia sapphirina (Diptera: Culicidae).

A novel cypovirus has been isolated from the mosquito Uranotaenia sapphirina (UsCPV) and shown to cause a chronic infection confined to the cytoplasm of epithelial cells of the gastric ceca and posterior stomach. The production of large numbers of virions and inclusion bodies and their arrangement into paracrystalline arrays gives the gut of infected insects a distinctive blue iridescence. The virions, which were examined by electron microscopy, are icosahedral (55 to 65 nm in diameter) with a central core that is surrounded by a single capsid layer. They are usually packaged individually within cubic inclusion bodies (polyhedra, approximately 100 nm across), although two to eight virus particles were sometimes occluded together. The virus was experimentally transmitted per os to several mosquito species. The transmission rate was enhanced by the presence of magnesium ions but was inhibited by calcium ions. Most of the infected larvae survived to adulthood, and the adults retained the infection. Electrophoretic analysis of the UsCPV genome segments (using 1% agarose gels) generated a migration pattern (electropherotype) that is different from those of the 16 Cypovirus species already recognized. UsCPV genome segment 10 (Seg-10) showed no significant nucleotide sequence similarity to the corresponding segment of the other cypoviruses that have previously been analyzed, and it has different "conserved" termini. A BLAST search of the UsCPV deduced amino acid sequence also showed little similarity to Antheraea mylitta CPV-4 (67 of 290 [23%]) or Choristoneura fumiferana CPV-16 (33 of 111 [29%]). We conclude that UsCPV should be recognized as a member of a new Cypovirus species (Cypovirus 17, strain UsCPV-17).

IA3, an aspartic proteinase inhibitor from Saccharomyces cerevisiae, is intrinsically unstructured in solution.

IA(3) is a highly specific and potent 68-amino acid endogenous inhibitor of yeast proteinase A (YprA), and X-ray crystallographic studies have shown that IA(3) binds to YprA as an alpha-helix [Li, M., Phylip, L. H., Lees, W. E., Winther, J. R., Dunn, B. M., Wlodawer, A., Kay, J., and Gustchina, A. (2000) Nat. Struct. Biol. 7, 113-117]. Surprisingly, only residues 2-32 of IA(3) are seen in the X-ray structure, and the remaining residues are believed to be disordered in the complex. We have used circular dichroism (CD) and nuclear magnetic resonance (NMR) spectroscopy to show that IA(3) is unstructured in the absence of YprA. Specifically, IA(3) produced a CD spectrum characteristic of an unstructured peptide, and the (15)N HSQC NMR spectra of IA(3) were characteristic of a polypeptide lacking intrinsic structure. We characterized the unstructured state of IA(3) by using singular-value decomposition (SVD) to analyze the CD data in the presence of TFE, by fully assigning the unbound IA(3) protein by NMR and comparing the chemical shifts to published random-coil values, and by measuring (1)H-(15)N heteronuclear NOEs, which are all consistent with an unfolded protein. The IA(3) samples used for NMR analyses were active and inhibited YprA with an inhibition constant (K(i)) of 1.7 nM, and the addition of YprA led to a large spectral transition in IA(3). Calorimetric (ITC) data also show that the overall enthalpy of the interaction between IA(3) and YprA is exothermic.