Coupling of gene regulation and carrier modification manipulates bacterial biofilms as robust living catalysts.

The biofilm of an engineered strain is limited by slow growth and low yield, resulting in an unsatisfactory ability to resist external stress and promote catalytic efficiency. Here, biofilms used as robust living catalysts were manipulated through dual functionalized gene regulation and carrier modification strategies. The results showed that gene overexpression regulates the autoinducer-2 activity, extracellular polymeric substance content and colony behavior of Escherichia coli, and the biofilm yield of csgD overexpressed strains increased by 79.35 % compared to that of the wild type strains (p < 0.05). In addition, the hydrophilicity of polyurethane fibres modified with potassium dichromate increased significantly, and biofilm adhesion increased by 105.80 %. Finally, the isoquercitrin yield in the catalytic reaction of the biofilm reinforced by the csgD overexpression strain and the modified carrier was 247.85 % higher than that of the untreated group. Overall, this study has developed engineered strains biofilm with special functions, providing possibilities for catalytic applications.

The Transcription Factor CsgD Contributes to Engineered Escherichia coli Resistance by Regulating Biofilm Formation and Stress Responses.

The high cell density, immobilization and stability of biofilms are ideal characteristics for bacteria in resisting antibiotic therapy. CsgD is a transcription activating factor that regulates the synthesis of curly fimbriae and cellulose in Escherichia coli, thereby enhancing bacterial adhesion and promoting biofilm formation. To investigate the role of CsgD in biofilm formation and stress resistance in bacteria, the csgD deletion mutant ΔcsgD was successfully constructed from the engineered strain E. coli BL21(DE3) using the CRISPR/Cas9 gene-editing system. The results demonstrated that the biofilm of ΔcsgD decreased by 70.07% (p < 0.05). Additionally, the mobility and adhesion of ΔcsgD were inhibited due to the decrease in curly fimbriae and extracellular polymeric substances. Furthermore, ΔcsgD exhibited a significantly decreased resistance to acid, alkali and osmotic stress conditions (p < 0.05). RNA-Seq results revealed 491 differentially expressed genes between the parent strain and ΔcsgD, with enrichment primarily observed in metabolism-related processes as well as cell membrane structure and catalytic activity categories. Moreover, CsgD influenced the expression of biofilm and stress response genes pgaA, motB, fimA, fimC, iraP, ompA, osmC, sufE and elaB, indicating that the CsgD participated in the resistance of E. coli by regulating the expression of biofilm and stress response. In brief, the transcription factor CsgD plays a key role in the stress resistance of E. coli, and is a potential target for treating and controlling biofilm.

Ability of near infrared spectroscopy to measure oxygenation in isolated upper extremity muscle compartments.

PURPOSE: Near infrared spectroscopy (NIRS), a noninvasive means for monitoring muscle oxygenation, may be useful in the diagnosis of acute compartment syndrome, a condition characterized by poor tissue perfusion. This study used the decrease in muscle oxygenation caused by exercise to investigate the ability of anatomic placement of NIRS sensor pads over compartments of the forearm to isolate perfusion values of a specific compartment. METHODS: We recruited 63 uninjured volunteers from a private clinic-based setting and placed NIRS sensor pads over the dorsal, volar, and mobile wad compartments of 1 forearm. A total of 49 participants also had the contralateral forearm monitored, which served as an internal control. Participants performed a series of 3 exercises designed to sequentially activate the muscles of each compartment. A washout period separated each exercise to allow perfusion to return to baseline. We compared NIRS values of each compartment recorded during muscle contraction with baseline values. RESULTS: Mean NIRS values decreased significantly from baseline during muscle contraction for all compartments, whereas mean NIRS values of muscle compartments that remained at rest showed little or no change. We observed no changes in NIRS values of the contralateral arm, which remained at rest during the entire data collection period. CONCLUSIONS: Although lack of an existing method for quantifying muscle perfusion precludes validation of this technique against a reference standard, this study suggests that NIRS can provide oxygenation values that are both sensitive and specific to muscle compartments of the forearm. Future studies should investigate NIRS among patients with upper extremity injuries. TYPE OF STUDY/LEVEL OF EVIDENCE: Diagnostic III.