Influence of Alternation of Sulfate Attack and Freeze-Thaw on Microstructure of Concrete

The effects of sulfate attack and freeze-thaw alternation on the concrete microstructure were systemically investigated by advanced test methods such as water absorption method, air void analysis, XRD, and SEM. The experimental results indicated that freeze-thaw damage is the major effective factor in the sulfate attack and freeze-thaw alternation test. In the alternation test, average aperture of capillary pores of specimens was smaller, pores uniformity was better, and water absorption rate was lower than those specimens used in the single freeze-thaw damage test. The average aperture and uniformity of pores could be improved by adding fly ash and slag. Damage was accumulated in many cycles of freeze-thaw and microcracks increased during the test. At the same time, the hydration products of the concrete developed into expansive gypsum, AFt, and TSA without any strength during sulfate attack. The results of the microstructure analysis form XRD and SEM are in accordance with that of AFt, about 3 μm length, around which other hydration products decomposed by C-S-H after sulfate attack resulted in loss of concrete strength

Cloning, over-expression, and characterization of a new carboxypeptidase A gene of Bacillus pumilus ML413 in Bacillus subtilis 168

Carboxypeptidase A (CPAs) are a well-studied group of zinc-containing exopeptidases that facilitate thebreakdown of proteins and peptides during metabolism. Carboxypeptidase A is typically produced in mammalian pancreatic, brain and other tissues. A new gene encoding carboxypeptidase A in the prokaryote Bacillus pumilus was amplified by polymerase chain reaction (PCR), ligated into the shuttle vector pMA5, and cloned in a GRAS bacteria-Bacillus subtilis 168 host. This gene sequence contained a 1621 bp open reading frame that encodes a protein of 540 amino acids. The optimum pH and temperature for enzyme activity were 7.5 and 50°C, respectively. The enzyme was quite stable at neutral pH and maintained about 65% activity following a 24 h incubation at 40°C. The Km of this CPA was 0.1 mM, much higher than in mammalian species. Glycerol, ammonium sulfate, and sodium citrate improved enzyme activity under optimal culture condition. The carboxypeptidase activity in recombinant B. subtilis 168 reached a maximum of 179 U ml-1 in a 5 L fermentator when cultured on improved medium. The over expression of  carboxypeptidase A in Bacillus subtilis has commercial applications.Key words: Bacillus pumilus, Bacillus subtilis 168, over-expression, orthogonal arrays, carboxypeptidase A,metallocarboxypeptidase

Properties and Microstructure of Roller Compacted Concrete With High Volume Low Quality Fly Ash

The properties of roller compacted concrete (RCC) with high dosage low quality fly ash are investigated, including strength, elastic modulus, ultimate tensile strain, drying shrinkage, autogenous deformation and durability, meanwhile the microstructure of the same paste containing low quality fly ash and ground low quality fly ash are studied, too. The properties of RCC containing 60% or more ground fly ash meet the design requirement. The microstructure is also tested by using X-ray diffraction (XRD), scanning electron microscopy (SEM), thermogravimetry-differential thermal analysis (TG-DTA) and Mercury Intrusion Porosimetry (MIP). The results indicate that ground fly ash plays the role of active component besides the physical filling effect at early age, while after 90 days, the surface of the glass beads is erroded and a lot of calcium silicate hydrate and calcium hydroxide staggered as storied dense structure. Low quality fly ash can accelerate the formation of hydration products, resulting in higher degree of cement hydration and denser microstructure, while the hydration heat in total is reduced. At the age of 90 days, fly ash has significant chemical activity and the properties of RCC will be improved at the later stage.DOI: http://dx.doi.org/10.5755/j01.ms.23.3.16318</p

Over-expression of Mycobacterium neoaurum 3-ketosteroid-\u3941-dehydrogenase in Corynebacterium crenatum for efficient bioconversion of 4-androstene-3,17-dione to androst-1,4-diene-3,17-dione

Background: 3-Ketosteroid-\u3941-dehydrogenase (KSDD), a flavoprotein enzyme, catalyzes the bioconversion of 4-androstene-3,17-dione (AD) to androst-1,4-diene-3,17-dione (ADD). To date, there has been no report about characterization of KSDD from Mycobacterium neoaurum strains, which were usually employed to produce AD or ADD by fermentation. Results: In this work, Corynebacterium crenatum was chosen as a new host for heterologous expression of KSDD from M. neoaurum JC-12 after codon optimization of the KSDD gene. SDS-PAGE and western blotting results indicated that the recombinant C. crenatum harboring the optimized ksdd (ksddII) gene showed significantly improved ability to express KSDD. The expression level of KSDD was about 1.6-fold increased C. crenatum after codon optimization. After purification of the protein, we first characterized KSDD from M. neoaurum JC-12, and the results showed that the optimum temperature and pH for KSDD activity were 30\ub0C and pH 7.0, respectively. The Km and Vmax values of purified KSDD were 8.91 \u3bcM and 6.43 mM/min. In this work, C. crenatum as a novel whole-cell catalyst was also employed and validated for bioconversion of AD to ADD. The highest transformation rate of AD to ADD by recombinant C. crenatum was about 83.87% after 10 h reaction time, which was more efficient than M. neoaurum JC-12 (only 3.56% at 10 h). Conclusions: In this work, basing on the codon optimization, overexpression, purification and characterization of KSDD, we constructed a novel system, the recombinant C. crenatum SYPA 5-5 expressing KSDD, to accumulate ADD from AD efficiently. This work provided new insights into strengthening sterol catabolism by overexpressing the key enzyme KSDD, for efficient ADD production

Circulating tumor DNA clearance predicts prognosis across treatment regimen in a large real-world longitudinally monitored advanced non-small cell lung cancer cohort

Background: Although growth advantage of certain clones would ultimately translate into a clinically visible disease progression, radiological imaging does not reflect clonal evolution at molecular level. Circulating tumor DNA (ctDNA), validated as a tool for mutation detection in lung cancer, could reflect dynamic molecular changes. We evaluated the utility of ctDNA as a predictive and a prognostic marker in disease monitoring of advanced non-small cell lung cancer (NSCLC) patients.Methods: This is a multicenter prospective cohort study. We performed capture-based ultra-deep sequencing on longitudinal plasma samples utilizing a panel consisting of 168 NSCLC-related genes on 949 advanced NSCLC patients with driver mutations to monitor treatment responses and disease progression. The correlations between ctDNA and progression-free survival (PFS)/overall survival (OS) were performed on 248 patients undergoing various treatments with the minimum of 2 ctDNA tests.Results: The results of this study revealed that higher ctDNA abundance (P=0.012) and mutation count (P=8.5x10(-4)) at baseline are associated with shorter OS. We also found that patients with ctDNA clearance, not just driver mutation clearance, at any point during the course of treatment were associated with longer PFS (P=2.2x10(-1)6, HR 0.28) and OS (P=4.5x10(-6), HR 0.19) regardless of type of treatment and evaluation schedule.Conclusions: This prospective real-world study shows that ctDNA clearance during treatment may serve as predictive and prognostic marker across a wide spectrum of treatment regimens

Investigation of Cement-Emulsified Asphalt in Plastic Concrete

The mechanical, mesodamage, and the microproperties of cement-emulsified asphalt concrete have been investigated by computed tomography (CT), scanning electron microscopy (SEM), X-ray diffraction (XRD), and thermogravimetric analysis (TG) in this work. Emulsified asphalt delayed the hydration of cement, making the early compressive strength of concrete develop slowly. However, the concrete compressive strength increased rapidly with the demulsification of emulsified asphalt. The damage stages of condense, expansion of volume, rapid crack propagation, and damage by real-time scanning have been observed. The CT mean value of the place near the lower end face suffered a larger decline but a smaller decline to the upper part of the sample. The evolution of concrete suffering damage to failure is a gradual development process, and no sharp expansion of brittle failure. The unhydrated cement, incorporation asphalt, fibrous C–S–H gel, CH, needle-shaped ettringite, and other hydration products were interwoven to constitute emulsified asphalt-cement paste, forming a spatial structure

EFFECTS OF MINERAL ADMIXTURE ON THE CARBONIC ACID LEACHING RESISTANCE OF CEMENT-BASED MATERIALS

In order to reveal the degradation process and deterioration mechanism of cement-based materials, this paper analyzes the effects of carbonic acid leaching on the mechanical strength of mortars, as well as relative mass loss, microstructure, and composition of various cement pastes. The results indicate that cement pastes containing less than 20 % fly ash have higher carbonic acid leaching resistance than cement pastes without fly ash. However, after carbonic acid leaching, the compressive strength of the samples with fly ash is lower than that of the cement pastes without fly ash. The leaching resistance is good for samples cured at an early age before leaching. Carbonic acid leaching proceeds from the paste surface to the interior. The incorporation of an appropriate amount of slag powder helps to increase the density of the paste. Due to the pozzolanic activity of fly ash at late-stage leaching, a mixture of fly ash (≤ 20 %) and slag powder (≤ 20 %) effectively improves carbonic acid leaching resistance. The products of early-stage leaching were mainly CaCO₃ and small amounts of SiO₂ and Fe₂O₃. The C-S-H phase at the paste surface suffered serious damage after long periods of leaching, and the main products of leaching were SiO₂ and Fe₂O₃

Over-expression of Mycobacterium neoaurum 3-ketosteroid-Δ1-dehydrogenase in Corynebacterium crenatum for efficient bioconversion of 4-androstene-3,17-dione to androst-1,4-diene-3,17-dione

Background: 3-Ketosteroid-Δ1-dehydrogenase (KSDD), a flavoprotein enzyme, catalyzes the bioconversion of 4-androstene-3,17-dione (AD) to androst-1,4-diene-3,17-dione (ADD). To date, there has been no report about characterization of KSDD from Mycobacterium neoaurum strains, which were usually employed to produce AD or ADD by fermentation. Results: In this work, Corynebacterium crenatum was chosen as a new host for heterologous expression of KSDD from M. neoaurum JC-12 after codon optimization of the KSDD gene. SDS-PAGE and western blotting results indicated that the recombinant C. crenatum harboring the optimized ksdd (ksddII) gene showed significantly improved ability to express KSDD. The expression level of KSDD was about 1.6-fold increased C. crenatum after codon optimization. After purification of the protein, we first characterized KSDD from M. neoaurum JC-12, and the results showed that the optimum temperature and pH for KSDD activity were 30°C and pH 7.0, respectively. The Km and Vmax values of purified KSDD were 8.91 μM and 6.43 mM/min. In this work, C. crenatum as a novel whole-cell catalyst was also employed and validated for bioconversion of AD to ADD. The highest transformation rate of AD to ADD by recombinant C. crenatum was about 83.87% after 10 h reaction time, which was more efficient than M. neoaurum JC-12 (only 3.56% at 10 h). Conclusions: In this work, basing on the codon optimization, overexpression, purification and characterization of KSDD, we constructed a novel system, the recombinant C. crenatum SYPA 5-5 expressing KSDD, to accumulate ADD from AD efficiently. This work provided new insights into strengthening sterol catabolism by overexpressing the key enzyme KSDD, for efficient ADD production

Influence of Inertia and Low Active Mineral Admixture on Strength and Microstructure of Cement-Based Materials

Cement-based materials were investigated by comparing the strength and microstructure of pastes and mortar containing limestone powder or low quality fly ash. The compressive strength of the mortar at 28 and 90 d was examined whose microstructures were characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), thermogravimetric analysis, and differential thermal analysis (TG-DTA). The results indicated that the strength of mortar decreased with increasing mineral admixtures. The limestone powder mainly acted as inert filler and hardly took part in the chemical reaction. Low quality fly ash may accelerate the formation of hydration products in samples with more chemically bonded water. This further resulted in a higher degree of cement hydration and denser microstructure, while the overall heat of hydration was reduced. At the early stage of hydration, low quality fly ash can be considered as an inert material whereas its reactivity at the later stage became high, especially for ground low quality fly ash