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A novel peptide containing antimicrobial sequence and gelatinase cleavage sites was designed for Staphylococcus aureus detection. Since Staphylococcus aureus could secrete gelatinase, the fluorescein labeled peptide GKRWWKWWRRPLGVRGC could be recognized and cleaved. The obtained products were able to be analyzed by capillary electrophoresis with fluorescence detection. To explore the effect of Staphylococcus aureus concentration on enzyme digestion ability of peptide, Staphylococcus aureus with different concentrations were incubated with the peptide. Results indicated that capillary electrophoretic method was efficient for determining Staphylococcus aureus content. Compared with traditional approaches for Staphylococcus aureus detection, capillary electrophoresis possessed higher efficiency, enhanced sensitivity, and low sample consumption. Moreover, the proposed peptide also presented desirable antimicrobial activity. It suggested that the novel antimicrobial peptide used in this research opens a new path of detecting Staphylococcus aureus by capillary electrophoretic method.

Chondroitin sulfate (CS) is a linear polysaccharide, which is widely used in medical, health care and other fields. Compared with the traditional animal tissue extraction method, microbial synthesis of CS has the advantages of controllability and easiness of scaling-up. In order to achieve an efficient synthesis of chondroitin sulfate A (CSA), we constructed a recombinant

Chondroitin sulfate (CS) is widely applied in the medical, clinical, and nutraceutical fields. However, all commercialized CS is extracted from animal tissues, which is associated with many problems, such as heavy reliance on antibiotics, animal excrement waste and long breeding time. Here, we developed a promising new green route for the de novo biosynthesis of non-animal CS from methanol, a one-carbon compound, using the engineered Pichia pastoris cell factory. Firstly, three exogenous genes, namely kfoC and kfoA from Escherichia coli K4 and tuaD from Bacillus subtilis, were introduced into P. pastoris to construct the chondroitin synthesis pathway, which yielded 5.5 mg L−1 chondroitin. After optimizing the expression of committed enzymes by codon optimization, the production of chondroitin improved by approximately 35-fold to 189.8 mg L−1. Thereafter, 6 kinds of Kozak sequences and 10 different endogenous promoters were compared and applied to achieve the expression of active chondroitin-4-O-sulfotransferase (41.3 U L−1). After integrating the chondroitin biosynthesis and sulfonation modules, the engineered strain, Pp006, successfully produced 182.0 mg L−1 CSA [GlcA-GalNAc (4S)] with a sulfation degree of 1.1%. We further improved the sulfation degree to 2.8% by strengthening the 3′-phosphoadenosine-5′-phosphosulfate (PAPS) supply with the overexpression of adenosine-5′-triphosphate sulfurylase and adenosine 5′-phosphosulfate kinase. In the fed-batch cultivation, the final titer of CSA reached 2.1 g L−1 and the sulfation degree increased to 4.0%. Moreover, the results suggest that the P. pastoris cell factory could be used as a green route to produce other sulfated glycosaminoglycans.

Multienzyme detection and monitoring enzyme activity in situ are significant for the disease to diagnose. This study aims to develop a quantum dots (QDs)-based nanoprobe Cyanine5-DDDLEVLFQFPGLVPRGSGGHHHHHH-QDs (Cy5-LEVLVP-QD), which is able to detect two enzymes inside a bent capillary using CE. Cy5-LEVLVP and QDs were allowed to bind with each other through metal affinity interaction and then injected the Cy5-LEVLVP-QD complex into a capillary with different bends, followed by related enzyme that can cleave the Cy5-LEVLVP peptide. The fluorescence of Cy5 was excited by QDs due to Förster resonance energy transfer. By monitoring the peaks produced by the original Cy5-LEVLVP-QD complex and a significant fluorescence change, sensitive analysis of two different enzymes was conducted. Therefore, the novel approach of using capillaries with semicircular bends could prove particularly useful for enzyme investigating in disease.

【Abstract】Aim To construct a prediction model based on functional mitral regurgitation (FMR) in patients with paroxysmal atrial fibrillation (PAF) for predicting atrial fibrillation recurrence after the post-circular pulmonary vein radiofrequency ablation (CPVA). Methods We retrospectively analyzed 289 patients with PAF who underwent CPVA for the first time . The patients were randomly divided into modeling group and verification group at the ratio of 75:25. In the modeling group, the multivariate logistic regression was used to analyze and construct a prediction model for post-CPVA recurrence in PAF patients, which was then validated in the verification group. Results (1) After 3-6 months of follow-up, the patients were divided into sinus rhythm group (252 cases) and recurrence group (24 cases); (2) In the modeling group, the age, LAD(left atrial diameter) and degree of MR (mild, moderate, severe) were higher in recurrence group than that of the sinus rhythm group, and the left atrial appendage emptying velocity (LAAV) was lower in recurrence group (all PP>0.05). Conclusion: The prediction model of atrial fibrillation (AF) recurrence after CPVA in PAF patients has good predictive efficacy, high specificity and accuracy.

The detection of protease activity in the body plays a significant role in the early diagnosis of diseases. However, enzymes inevitably come into contact with various complex biological fluids in the body during the flow, which greatly increases the detection difficulty. Therefore, protease detection in vivo has great challenges. Herein, we report a new assay for detecting protease using capillary electrophoresis inside a capillary with semicircular bends. We first designed a peptide substrate, and then the peptide was self-assembled with quantum dots to form a QDs-peptide substrate. The capillary was bent to semicircular-shaped turns and served as a micro-reactor to allow protease and substrate react in it. Due to the different electrophoretic velocity, the protease and the substrate were mixed inside the bent capillary with sequential injections and the cleavage of the substrate can be detected using capillary electrophoresis combined with Forster resonance energy transfer technology. This novel assay will greatly expand the detection of enzyme activity in vivo.

This paper studies the problem of determining the optimal initial stock level of recoverable components and the aircraft preventive maintenance inspection interval for an aircraft fleet. Demand for spare components arises from both corrective and preventive maintenance tasks. An aircraft that experiences failures while performing a mission is maintained by replacing failed components with working spares. In order to reduce the amount of this unplanned, corrective maintenance that is performed, aircraft are usually inspected periodically and components found to be in poor condition replaced preventively. The supply of spares is critical to maintaining fleet operational requirements but determining the stock required while considering critical fleet operational requirements and budget limitations is a complex task. When spares are unavailable, inoperative aircraft must wait for failed components to be repaired. In such situations, cannibalisation may be performed, where working components are remove...

Cannibalisation refers to a maintenance action where an unserviceable part in an inoperative platform is replaced by a serviceable part of the same type from another platform. It helps a fleet meet operational requirements when spares are in short supply but leads to more maintenance tasks to be carried out. In practice, cannibalisation may be performed in an unrestricted manner, or through the use of cannibalisation birds. A cannibalisation bird is a platform which is selected as the primary source of cannibalisation, while any inoperative platform can be a cannibalisation source under the unrestricted policy. In order to aid fleet managers in making cannibalisation-related decisions, this paper presents a hierarchical coloured Petri net (HCPN) model of a fleet operation and maintenance process which considers mission-oriented operation, multiple level maintenance, multiple cannibalisation policies (no cannibalisation, unrestricted cannibalisation and cannibalisation bird), maintenance scheduling and spare inventory management. The model is applied to an example fleet to compare the effects of different cannibalisation policies on fleet performance using a number of performance measures related to reliability and maintenance and to optimise the number of cannibalisation birds used and the length of time that a platform is taken as a cannibalisation bird for the fleet.