Computational design of CRISPR guide RNAs to enable strain-specific control of microbial consortia.

Microbes naturally coexist in complex, multistrain communities. However, extracting individual microbes from and specifically manipulating the composition of these consortia remain challenging. The sequence-specific nature of CRISPR guide RNAs can be leveraged to accurately differentiate microorganisms and facilitate the creation of tools that can achieve these tasks. We developed a computational program, ssCRISPR, which designs strain-specific CRISPR guide RNA sequences with user-specified target strains, protected strains, and guide RNA properties. We experimentally verify the accuracy of the strain specificity predictions in both Escherichia coli and Pseudomonas spp. and show that up to three nucleotide mismatches are often required to ensure perfect specificity. To demonstrate the functionality of ssCRISPR, we apply computationally designed CRISPR-Cas9 guide RNAs to two applications: the purification of specific microbes through one- and two-plasmid transformation workflows and the targeted removal of specific microbes using DNA-loaded liposomes. For strain purification, we utilize gRNAs designed to target and kill all microbes in a consortium except the specific microbe to be isolated. For strain elimination, we utilize gRNAs designed to target only the unwanted microbe while protecting all other strains in the community. ssCRISPR will be of use in diverse microbiota engineering applications.

Implementation and delivery of electronic health records training programs for nurses working in inpatient settings: a scoping review.

OBJECTIVES: Well-designed electronic health records (EHRs) training programs for clinical practice are known to be valuable. Training programs should be role-specific and there is a need to identify key implementation factors of EHR training programs for nurses. This scoping review (1) characterizes the EHR training programs used and (2) identifies their implementation facilitators and barriers. MATERIALS AND METHODS: We searched MEDLINE, CINAHL, PsycINFO, and Web of Science on September 3, 2023, for peer-reviewed articles that described EHR training program implementation or delivery to nurses in inpatient settings without any date restrictions. We mapped implementation factors to the Consolidated Framework for Implementation Research. Additional themes were inductively identified by reviewing these findings. RESULTS: This review included 30 articles. Healthcare systems' approaches to implementing and delivering EHR training programs were highly varied. For implementation factors, we observed themes in innovation (eg, ability to practice EHR skills after training is over, personalizing training, training pace), inner setting (eg, availability of computers, clear documentation requirements and expectations), individual (eg, computer literacy, learning preferences), and implementation process (eg, trainers and support staff hold nursing backgrounds, establishing process for dissemination of EHR updates). No themes in the outer setting were observed. DISCUSSION: We found that multilevel factors can influence the implementation and delivery of EHR training programs for inpatient nurses. Several areas for future research were identified, such as evaluating nurse preceptorship models and developing training programs for ongoing EHR training (eg, in response to new EHR workflows or features). CONCLUSIONS: This scoping review highlighted numerous factors pertaining to training interventions, healthcare systems, and implementation approaches. Meanwhile, it is unclear how external factors outside of a healthcare system influence EHR training programs. Additional studies are needed that focus on EHR retraining programs, comparing outcomes of different training models, and how to effectively disseminate updates with the EHR to nurses.

Hemostatic patch with ultra-strengthened mechanical properties for efficient adhesion to wet surfaces.

Controlling traumatic bleeding from damaged internal organs while effectively sealing the wound is critical for saving the lives of patients. Existing bioadhesives suffer from blood incompatibility, insufficient adhesion to wet surfaces, weak mechanical properties, and complex application procedures. Here, we engineered a ready-to-use hemostatic bioadhesive with ultra-strengthened mechanical properties and fatigue resistance, robust adhesion to wet tissues within a few seconds of gentle pressing, deformability to accommodate physiological function and action, and the ability to stop bleeding efficiently. The engineered hydrogel, which demonstrated high elasticity (>900%) and toughness (>4600 kJ/m3), was formed by fine-tuning a series of molecular interactions and crosslinking mechanisms involving N-hydroxysuccinimide (NHS) conjugated alginate (Alg-NHS), poly (ethylene glycol) diacrylate (PEGDA), tannic acid (TA), and Fe3+ ions. Dual adhesive moieties including mussel-inspired pyrogallol/catechol and NHS synergistically enhanced wet tissue adhesion (>400 kPa in a wound closure test). In conjunction with physical sealing, the high affinity of TA/Fe3+ for blood could further augment hemostasis. The engineered bioadhesive demonstrated excellent in vitro and in vivo biocompatibility as well as improved hemostatic efficacy as compared to commercial Surgicel®. Overall, the hydrogel design strategy described herein holds great promise for overcoming existing obstacles impeding clinical translation of engineered hemostatic bioadhesives.

Engineering E. coli strains using antibiotic-resistance-gene-free plasmids.

We created a generalizable pipeline for antibiotic-resistance-gene-free plasmid (ARGFP)-based cloning using a dual auxotrophic- and essential-gene-based selection strategy. We use auxotrophic selection to construct plasmids in engineered E. coli DH10B cloning strains and both auxotrophic- and essential-gene-based selection to (1) select for recombinant strains and (2) maintain a plasmid in E. coli Nissle 1917, a common chassis for engineered probiotic applications, and E. coli MG1655, the laboratory "wild-type" E. coli strain. We show that our approach has comparable efficiency to that of antibiotic-resistance-gene-based cloning. We also show that the double-knockout Nissle and MG1655 strains are simple to transform with plasmids of interest. Notably, we show that the engineered Nissle strains are amenable to long-term plasmid maintenance in repeated culturing as well as in the mouse gut, demonstrating the potential for broad applications while minimizing the risk of antibiotic resistance spread via horizontal gene transfer.