Electroporation of SUMO-His-Cre protein triggers a specific recombinase-mediated cassette exchange in HEK 293T cells.

Cre recombinase is a widely-used genetic manipulation of genomic DNA. However, the conventional transfection of the DNA vectors expressing the Cre recombinase or viral transduction method yields low transfection efficiencies or insertion mutagenesis. The present paper evaluated whether the direct protein delivery of Cre recombinase through electroporation can induce the Cre-mediated recombination in the HEK 293T cells. Here, the small ubiquitin-related modifier (SUMO) -tagged His-Cre fusion protein was expressed in a soluble pattern in the Eschrichria coli (E.coli) cells, purified using affinity chromatography, and finally electroporated into the HEK 293T cells. These cells were previously transfected with three different Cre reporter vectors. The electroporation of the HEK 293T cells revealed either the activation of EGFP expression, or a decrease in RFP expression, and a concomitant increase in EGFP expression, indicating a desired recombinase-mediated cassette exchange (RMCE) event (conversion of RFP to EGFP), and a biological activity of the purified SUMO-His-Cre protein. The fusion protein is expected to serve in the Easi-CRISPR-LoxP-mediated genome editing to generate transgenic animal models.

Fermentation for future food systems: Precision fermentation can complement the scope and applications of traditional fermentation.

Modern biotechnology holds great potential for expanding the scope of fermentation to create novel foods and improve the sustainability of food production.

Early Mobilization of Mild-Moderate Intracerebral Hemorrhage Patients in a Stroke Center: A Randomized Controlled Trial.

Background. Few studies have addressed early out-of-bed mobilization specifically in acute intracerebral hemorrhage (ICH) patients. Patient benefit in such cases is unclear, with early intervention timing and duration identical to those in standard care. Objective. We investigated the efficacy of an early mobilization (EM) protocol, administered within 24 to 72 hours of stroke onset, for early functional independence in mild-moderate ICH patients. Methods. Sixty patients admitted to a stroke center within 24 hours of ICH were randomly assigned to early mobilization (EM) or standard early rehabilitation (SER). The EM group underwent an early out-of-bed mobilization protocol, while the SER group underwent a standard protocol focusing on in-bed training in the stroke center. Intervention in both groups lasted 30 minutes per session, once a day, 5 days a week. Motor subscales of the Functional Independence Measure (FIM-motor; primary outcome), Postural Assessment Scale for Stroke Patients, and Functional Ambulation Category (FAC) were evaluated (assessor-blinded) at baseline, and at 2 weeks, 4 weeks, and 3 months after stroke. Length of stay in the stroke center was also recorded. Results. The EM group showed significant improvement in FIM-motor score at all evaluated time points (P = .004) and in FAC outcomes at 2 weeks (P = .033) and 4 weeks (P = .011) after stroke. Length of stay in the stroke center was significantly shorter for the EM group (P = .004). Conclusion. Early out-of-bed mobilization via rehabilitation in a stroke center, within 24 to 72 hours of ICH, may improve early functional independence compared with standard early rehabilitation. Clinical Trial Registration: NCT03292211.

Development of an Objective Portable Measurement Device for Spinal Joint Accessory Motion Testing.

Joint accessory motion testing (JAMT) is a standard procedure used by manual therapists to assess and treat musculoskeletal disorders. Joint accessory motion (JAM) is movement that occurs between joint surfaces, and can be induced by applying force. The motion amount, end feel, symptoms, and resistance perceived by therapists during test procedures are recorded as evidence for the diagnosis, prognosis, treatment decision making, and intervention outcome. However, previous studies have shown that accessory motion tests have insufficient reliability. Recently, many instruments have been developed to increase test reliability, but these instruments quantify the test results with a single probe and utilize the external environment as a reference. Therefore, the measured displacement amount may be affected by other spinal segments. This study proposes an objective portable measurement device with two indenter probes for spinal JAMT, wherein the JAM was quantified by displacement and force measurements between two bones. The instrument was verified with a homemade spinal simulator and computer simulation. The results showed that the force-displacement curves measured by the JAMT device (JAMTD) and those simulated by the computer model exhibited similar characteristics. Moreover, a two-probe measurement could distinguish the differences in stiffness better than a one-probe measurement.

Fall Risk Program for Oncology Inpatients: Addition of the "Traffic Light" Fall Risk Assessment Tool.

BACKGROUND: The incidence of falls on inpatient oncology units indicated the need for quality improvement. This project aimed to reduce falls by implementing a fall reduction plan including the "Traffic Light" Fall Risk Assessment Tool (TL-FRAT). LOCAL PROBLEM: We retrospectively reviewed the oncology unit fall data from January 2013 to September 2014 and found that the average fall incidence was high. METHODS: The project used a program evaluation design, and the process was guided by Kotter's 8-step change model. INTERVENTIONS: We implemented the TL-FRAT to classify oncology inpatients at a high risk of falling in advance. RESULTS: The average fall incidence and falls with injury during the project were reduced. CONCLUSIONS: Adding the TL-FRAT to the fall protocol on the units effectively reduced the incidence of falls related to impaired mobility. The TL-FRAT can improve nurses' sensitivity to falls related to impaired mobility and, subsequently, guide corresponding fall prevention strategies.

Reliability of lower extremity muscle strength measurements with handheld dynamometry in stroke patients during the acute phase: a pilot reliability study.

[Purpose] No literature has described a suitable method for measuring muscle strength in a supine position during acute phase after stroke. This study investigated the feasibility and reliability of using a commercial handheld dynamometer to measure the muscle strengths of the hip flexor, knee extensor, and dorsiflexor in the supine position with a modified method for patients at a stroke intensive care center within 7 days of stroke onset. [Subjects and Methods] Fifteen persons with acute stroke participated in this cross-sectional study. For each patient, the muscle strengths of the hip flexors, knee extensors, and dorsiflexors were measured twice by two testers on the same day. Each patient was re-tested at the same time of day one day later. Inter-rater and test-retest reliability were then determined by the intraclass correlation coefficients (ICCs). [Results] For the three muscle groups, the inter-rater reliability ICCs were all 0.99 and the test-retest reliability ICCs were greater than 0.85. The investigated method thus has good inter-rater reliability and high agreement between the test-retest measurements, with acceptable measurement errors. [Conclusion] The modified method using a handheld dynamometer to test the muscle strength of acute stroke patients is a feasible and reliable method for clinical use.

Curcumin-mediated lifespan extension in Caenorhabditis elegans.

Curcumin is the active ingredient in the herbal medicine and dietary spice, turmeric (Curcuma longa). It has a wide range of biological activities, including anti-inflammatory, antioxidant, chemopreventive, and chemotherapeutic activities. We examined the effects of curcumin on the lifespan and aging in Caenorhabditis elegans, and found that it responded to curcumin with an increased lifespan and reduced intracellular reactive oxygen species and lipofuscin during aging. We analyzed factors that might influence lifespan extension by curcumin. We showed that lifespan extension by curcumin in C. elegans is attributed to its antioxidative properties but not its antimicrobial properties. Moreover, we showed that lifespan extension had effects on body size and the pharyngeal pumping rate but not on reproduction. Finally, lifespan tests with selected stress- and lifespan-relevant mutant strains revealed that the lifespan-extending phenotype was absent from the osr-1, sek-1, mek-1, skn-1, unc-43, sir-2.1, and age-1 mutants, whereas curcumin treatment prolonged the lifespan of mev-1 and daf-16 mutants. Our study has unraveled a diversity of modes of action and signaling pathways to longevity and aging with curcumin exposure in vivo.

Biomolecular pathway databases.

From the database point of view, biomolecular pathways are sets of proteins and other biomacromolecules that represent spatio-temporally organized cascades of interactions with the involvement of low-molecular compounds and are responsible for achieving specific phenotypic biological outcomes. A pathway is usually associated with certain subcellular compartments. In this chapter, we analyze the major public biomolecular pathway databases. Special attention is paid to database scope, completeness, issues of annotation reliability, and pathway classification. In addition, systems for information retrieval, tools for mapping user-defined gene sets onto the information in pathway databases, and their typical research applications are reviewed. Whereas today, pathway databases contain almost exclusively qualitative information, the desired trend is toward quantitative description of interactions and reactions in pathways, which will gradually enable predictive modeling and transform the pathway databases into analytical workbenches.

Databases of protein-protein interactions and complexes.

In the current understanding, translation of genomic sequences into proteins is the most important path for realization of genome information. In exercising their intended function, proteins work together through various forms of direct (physical) or indirect interaction mechanisms. For a variety of basic functions, many proteins form a large complex representing a molecular machine or a macromolecular super-structural building block. After several high-throughput techniques for detection of protein-protein interactions had matured, protein interaction data became available in a large scale and curated databases for protein-protein interactions (PPIs) are a new necessity for efficient research. Here, their scope, annotation quality, and retrieval tools are reviewed. In addition, attention is paid to portals that provide unified access to a variety of such databases with added annotation value.

Gastric Cancer (Biomarkers) Knowledgebase (GCBKB): A Curated and Fully Integrated Knowledgebase of Putative Biomarkers Related to Gastric Cancer.

The Gastric Cancer (Biomarkers) Knowledgebase (GCBKB) (http://biomarkers.bii.a-star.edu.sg/background/gastricCancerBiomarkersKb.php) is a curated and fully integrated knowledgebase that provides data relating to putative biomarkers that may be used in the diagnosis and prognosis of gastric cancer. It is freely available to all users. The data contained in the knowledgebase was derived from a large literature source and the putative biomarkers therein have been annotated with data from the public domain. The knowledgebase is maintained by a curation team who update the data from a defined source. As well as mining data from the literature, the knowledgebase will also be populated with unpublished experimental data from investigators working in the gastric cancer biomarker discovery field. Users can perform searches to identify potential markers defined by experiment type, tissue type and disease state. Search results may be saved, manipulated and retrieved at a later date. As far as the authors are aware this is the first open access database dedicated to the discovery and investigation of gastric cancer biomarkers.