Whole-cell biocatalyst for cadaverine production using stable, constitutive and high expression of lysine decarboxylase in recombinant Escherichia coli W3110.

Microbial production of industrial chemicals is a sustainable approach to reduce the dependence on petroleum-based chemicals such as acids, alcohols, and amines, in which the cadaverine is a natural diamide and serves as one of the key monomers for biopolymer production. In this study, the constitutive promoter J23100 driven lysine decarboxylase (CadA) for cadaverine production was established and compared in different Escherichia coli strains. The best chassis designed as JW, expressed the highest amount of CadA by using J23100 promoter, showing stable and high copy numbers (i.e., PCN > 100) when culture in the antibiotic-free medium. JW attained a CadA activity of 167 g-DAP/g-DCW-h and had the maximum biocatalyst of 45.6 g-DCW/L in fed-batch fermentation. In addition, JW was able to convert 2.5 M L-lysine to 221 g/L cadaverine, with 86 % yield and 55.3 g/L-h productivity. The whole-cell biocatalyst could be reused over four times at an average of 97 % conversion when supplied half of fresh cells in the reaction. This work developed a stable, constitutive expression, long-term preservation, high-level expression of CadA for DAP production, and paved an alternative opportunity of bio-nylon for industry in the future.

A nanopatterned cell-seeded cardiac patch prevents electro-uncoupling and improves the therapeutic efficacy of cardiac repair.

The heart is an extremely sophisticated organ with nanoscale anisotropic structure, contractility and electro-conductivity; however, few studies have addressed the influence of cardiac anisotropy on cell transplantation for myocardial repair. Here, we hypothesized that a graft's anisotropy of myofiber orientation determines the mechano-electrical characteristics and the therapeutic efficacy. We developed aligned- and random-orientated nanofibrous electrospun patches (aEP and rEP, respectively) with or without seeding of cardiomyocytes (CMs) and endothelial cells (ECs) to test this hypothesis. Atomic force microscopy showed a better beating frequency and amplitude of CMs when cultured on aEP than that from cells cultured on rEP. For the in vivo test, a total of 66 rats were divided into six groups: sham, myocardial infarction (MI), MI + aEP, MI + rEP, MI + CM-EC/aEP and MI + CM-EC/rEP (n ≥ 10 for each group). Implantation of aEP or rEP provided mechanical support and thus retarded functional aggravation at 56 days after MI. Importantly, CM-EC/aEP implantation further improved therapeutic outcomes, while cardiac deterioration occurred on the CM-EC/rEP group. Similar results were shown by hemodynamic and infarct size examination. Another independent in vivo study was performed and electrocardiography and optical mapping demonstrated that there were more ectopic activities and defective electro-coupling after CM-EC/rEP implantation, which worsened cardiac functions. Together these results provide comprehensive functional characterizations and demonstrate the therapeutic efficacy of a nanopatterned anisotropic cardiac patch. Importantly, the study confirms the significance of cardiac anisotropy recapitulation in myocardial tissue engineering, which is valuable for the future development of translational nanomedicine.

Pharmacokinetic and pharmacodynamic properties of batifiban coadministered with antithrombin agents in Chinese healthy volunteers.

The combined use of batifiban, a synthetic platelet GPII b/ IIIa receptor antagonist, and antithrombin agents is an attractive option for the treatment of patients with non-ST-segment elevation (NSTE) acute coronary syndrome (ACS) and those scheduled for percutaneous coronary intervention. To observe whether antithrombin agents affect the pharmacokinetic and pharmacodynamic properties of batifiban in combination therapy and optimize clinical administration dosage of batifiban, an open-label and parallel study was conducted. Thirty healthy subjects were randomly divided into three groups, which were sequentially treated with batifiban alone, or oral coadministration of clopidogrel, aspirin and UFH, or batifiban coadministered with these antithrombin agents. Blood samples were collected at pre-specified time points. The evaluation index included the inhibition of platelet aggregation and pharmacokinetic parameters. The pharmacokinetic parameters of batifiban and batifiban coadministered with antithrombin agents showed no significant differences. The mean inhibition rate of platelet aggregation (%) suggested that neither batifiban alone nor antithrombin agents alone could provide such potent inhibition rate (>80%) to obtain the best clinical efficacy, but they had a synergistic effect on platelet inhibition. No serious adverse effects were observed. The results in these healthy subjects suggest that batifiban coadministrated with antithrombin agents could achieve optimum clinical treatment effect for patients with NSTE ACS, and also those scheduled for percutaneous coronary intervention.

Bioactive collagen-grafted poly-L-lactic acid nanofibrous membrane for cartilage tissue engineering.

Biodegradable nanofibrous membrane was prepared from poly-L-lactic acid by electrospinning and used as a scaffold for cartilage tissue engineering. Operating parameters during the electrospinning process were studied in terms of their influences on fiber diameter, membrane porosity and pore size. In order to improve cell attachment and growth, nanofibrous membrane was subject to DC-pulsed oxygen plasma treatment, followed by acrylic acid grafting and collagen coating by covalent binding of collagen to carboxylic moieties of the polyacrylic acid. The membrane was fully characterized for its physical and chemical properties. Primary chondrocyte cells seeded into the membrane proliferated well and maintain high viability within the membrane from Confocal Laser Scanning Microscopy. Cell differentiation was confirmed by secretion of glycoaminoglycan and collagen during the cultivation period. Scanning electron microscope observation of the cell-scaffold construct confirms the tight attachment of cells to nanofibers and in-growth of cells into the interior of the membrane with proper maintenance of morphology and structure of chondrocytes.

[Analysis on relationship between regional economic development and sewage disposal].

Based on the relationship between district GDP and sewage disposal, the water environment protection effect in 3 cities, Suzhou, Nanjing and Xuzhou, with different economic development degrees in Jiangsu Province was dynamically analyzed. The economy in Suzhou was well developed, where the foreign capital proportion was in a high level. Its GDP per capita was 53,800 yuan in 2005 and the sewage disposal grew linearly when its GDP increased in the study time period. Nanjing was less developed than Suzhou, and the state-owned enterprises in large and medium sizes were in a high percentage. Its GDP per capita was 37,100 yuan in 2005, while the sewage disposal reduced linearly when its GDP increased in the study time period. The economy in Xuzhou is under-developed, where coal-based heavy industry was the most important one. The GDP per capita in this city was 13,200 yuan in 2005 and the sewage disposal fluctuated when its GDP increased in the study time period. According to the relationship between economic development and sewage disposal in different cities, we suggested that the water environment protection in Suzhou should focus on the control of both water pollutant total emission and emission concentration, the major work in Nanjing should focus on adjusting the industrial structure and meanwhile controlling the total emission of water pollutants, while in Xuzhou the water pollutant emission concentration should be firstly controlled.