ABSTRACT
As the span of concrete-filled steel tube (CFST) arch bridges increases, the hydration heat temperature effect of concrete inside steel tube becomes more severe, which increases the safety risk during the construction process. Therefore, a numerical simulation of the mechanical response of a long-span CFST arch bridge under the influence of hydration heat was carried out. First, based on the hydration heat conduction theory, a finite element model of the transient temperature field of a CFST arch rib was established. The temperature distribution of the CFST arch rib and its variation with time were revealed, and an approximate formula for the distribution of the hydration heat temperature along the radial direction of the CFST was provided. Subsequently, the variation law of the thermal stress of a CFST during hydration heat release was investigated. Finally, based on the principle of temperature equivalence, a finite element model of the overall CFST arch rib was established to examine the effect of hydration heat on the deformation of the arch rib. The results reveal that the hydration heat temperature field of the CFST arch rib exhibits nonlinear and axisymmetric characteristics. The maximum temperature of the section and the maximum temperature difference can reach 73.5 °C and 33.2 °C, respectively. Because of the influence of the hydration heat, there is a significant stress gradient in the cross section of the arch rib. A maximum radial stress of 2.08 MPa is attained, indicating a risk of concrete cracking. Additionally, the displacement along the transverse and vertical directions of the chord tube exhibits an initial increase, followed by a decrease over time. The maximum transverse displacement of the chord tube reaches 70.6 mm, while the vertical displacement reaches 117.8 mm.
ABSTRACT
Bacillopeptidase F (Bpr) is a fibrinolytic serine protease produced by Bacillus subtilis. Its precursor is composed of a signal peptide, an N-terminal propeptide, a catalytic domain, and a long C-terminal extension (CTE). Several active forms of Bpr have been previously reported, but little is known about the maturation of this enzyme. Here, a gene encoding a Bpr (BprL) was cloned from B. subtilis LZW and expressed in B. subtilis WB700, and three fibrinolytic mature forms with apparent molecular masses of 45, 75, and 85 kDa were identified in the culture supernatant. After treatment with urea, the 75-kDa mature form had the same molecular mass as the 85-kDa mature form, from which we infer that they adopt different conformations. Mutational analysis revealed that while the 85-kDa mature form is generated via heterocatalytic processing of a BprL proform by an unidentified protease of B. subtilis, the production of the 75- and 45-kDa mature forms involves both hetero- and autocatalytic events. From in vitro analysis of BprL and its sequential C-terminal truncation variants, it appears that partial removal of the CTE is required for the initiation of autoprocessing of the N-terminal propeptide, which is composed of a core domain (N*) and a 15-residue linker peptide, thereby yielding the 45-kDa mature form. These data suggest that the differential processing of BprL, either heterocatalytically or autocatalytically, leads to the formation of multiple mature forms with different molecular masses or conformations.
Subject(s)
Bacillus subtilis/enzymology , Serine Endopeptidases/chemistry , Serine Endopeptidases/metabolism , Soil Microbiology , Amino Acid Sequence , Bacillus subtilis/drug effects , Bacillus subtilis/genetics , Bacillus subtilis/physiology , Biocatalysis , Catalytic Domain , Cloning, Molecular , Escherichia coli/genetics , Fibrinolysis , Molecular Sequence Data , Mutation , Peptides/metabolism , Protein Folding , Protein Modification, Translational , Serine Endopeptidases/genetics , Urea/pharmacologyABSTRACT
The incorporation of the structural elements of thermostable enzymes into their less stable counterparts is generally used to improve enzyme thermostability. However, the process of engineering enzymes with both high thermostability and high activity remains an important challenge. Here, we report that the thermostability and activity of a thermophilic subtilase were simultaneously improved by incorporating structural elements of a psychrophilic subtilase. There were 64 variable regions/residues (VRs) in the alignment of the thermophilic WF146 protease, mesophilic sphericase, and psychrophilic S41. The WF146 protease was subjected to systematic mutagenesis, in which each of its VRs was replaced with those from S41 and sphericase. After successive rounds of combination and screening, we constructed the variant PBL5X with eight amino acid residues from S41. The half-life of PBL5X at 85°C (57.1 min) was approximately 9-fold longer than that of the wild-type (WT) WF146 protease (6.3 min). The substitutions also led to an increase in the apparent thermal denaturation midpoint temperature (Tm) of the enzyme by 5.5°C, as determined by differential scanning calorimetry. Compared to the WT, PBL5X exhibited high caseinolytic activity (25 to 95°C) and high values of Km and kcat (25 to 80°C). Our study may provide a rational basis for developing highly stable and active enzymes, which are highly desired in industrial applications.
Subject(s)
Serine Endopeptidases/chemistry , Serine Endopeptidases/metabolism , Amino Acid Sequence , Crystallography, X-Ray , Enzyme Stability/genetics , Hot Temperature , Kinetics , Models, Molecular , Molecular Sequence Data , Mutagenesis, Site-Directed , Sequence Alignment , Serine Endopeptidases/geneticsABSTRACT
There is often great uncertainty in water quality modeling for urban drainage systems because water quality variation in systems is complex and affected by many factors. The stormwater management model (SWMM) was applied to a small-scale urban catchment with a simple and well-maintained stormwater drainage system without illicit connections. This was done to assess uncertainty in build-up and wash-off modeling of pollutants within the generalized likelihood uncertainty estimation (GLUE) methodology, based on a well-calibrated water quantity model. The results indicated great uncertainty of water quality modeling within the GLUE methodology. Comparison of uncertainties in various pollutant build-up and wash-off models that were available in SWMM indicated that those uncertainties varied slightly. This may be a consequence of the specific characteristics of rainfall events and experimental sites used in the study. The uncertainty analysis of water quality parameters in SWMM is conducive to effectively evaluating model reliability, and provides an experience base for similar research and applications.
Subject(s)
Environmental Monitoring/methods , Models, Theoretical , Water Pollution, Chemical/analysis , Water Quality , China , Rain , UncertaintyABSTRACT
Pollutant characteristics of surface runoff, sanitary sewage and sewer deposits from a combined sewer system in Shanghai were studied. It was confirmed that the content of Zn, NH4(+) -N, P, respectively in surface runoff, sanitary sewage and sewer deposits was specific and relatively stable based on contrasting pollutant characteristics of different sources. Three non-dimensional parameters-Zn/P, NH4(+) -N/Zn, P/K were proposed as characteristic index for surface runoff, sanitary sewage and sewer deposits. In order to find sources of pollutants in CSO, the application of chemical mass balance method was discussed: the average contributions to CSO were 42.8% for surface runoff, 12.2% for sanitary sewage, 23.8% for sewer deposits, and the results basically reflected the composition of CSO pollution. The research method and results can provide guidance for pollutant source apportionment of combined sewer overflows.
Subject(s)
Sewage/analysis , Water Pollution/analysis , ChinaABSTRACT
Curcumin is a major active component of Curcuma aromatica salisb, which has been shown to inhibit proliferation of a wide variety of tumor cells. In this study, the molecular mechanisms of curcumin inducing apoptosis in human hepatoma SMMC-7721 cells were examined. We find that curcumin inhibits the growth of SMMC-7721 cells significantly in a concentration-depenent manner, with typical apoptotic morphological changes of cellular nuclei. Annexin-V/PI double staining detected by flow cytometry and expression of the relative apoptotic proteins (Bax, Bcl-2 and caspase-3) revealed a strong apoptosis-inducing competent of curcumin in SMMC-7721 cells. Curcumin increased the expression of bax protein while decreasing that of bc1-2 protein significantly. The results suggest that curcumin induction of apoptosis involves modulation of bax/bcl-2 in SMMC-7721 cells and provide a molecular basis for the development of naturally compounds as novel anticancer agents for human hepatomas.