Ultrasound-mediated spatial and temporal control of engineered cells in vivo.

Remote regulation of cells in deep tissue remains a significant challenge. Low-intensity pulsed ultrasound offers promise for in vivo therapies due to its non-invasive nature and precise control. This study uses pulsed ultrasound to control calcium influx in mammalian cells and engineers a therapeutic cellular device responsive to acoustic stimulation in deep tissue without overexpressing calcium channels or gas vesicles. Pulsed ultrasound parameters are established to induce calcium influx in HEK293 cells. Additionally, cells are engineered to express a designed calcium-responsive transcription factor controlling the expression of a selected therapeutic gene, constituting a therapeutic cellular device. The engineered sonogenetic system's functionality is demonstrated in vivo in mice, where an implanted anti-inflammatory cytokine-producing cellular device effectively alleviates acute colitis, as shown by improved colonic morphology and histopathology. This approach provides a powerful tool for precise, localized control of engineered cells in deep tissue, showcasing its potential for targeted therapeutic delivery.

Protein Gas Vesicles of Bacillus megaterium as Enhancers of Ultrasound-Induced Transcriptional Regulation.

Gas vesicles (GVs) are large cylindrical gas-filled protein assemblies found in diverse aquatic bacteria that enable their adaptation of buoyancy. GVs have already been used as ultrasound contrasting agents. Here, we investigate GVs derived from Bacillus megaterium, aiming to minimize the number of accessory Gvps within the GV gene cluster and demonstrate the use of GVs as enhancers of acoustic radiation force administered by ultrasound. Three (GvpR, GvpT, and GvpU) out of 11 genes in the cluster were found to be dispensable for functional GV formation, and their omission resulted in narrower GVs. Two essential proteins GvpJ and GvpN were absent from recently determined GV structures, but GvpJ was nevertheless found to be tightly bound to the cylindrical part of GVs in this study. Additionally, the N-terminus of GvpN was observed to play an important role in the formation of mature GVs. The binding of engineered GvpC fromAnabaena flos-aquae to HEK293 cells via integrins enhanced the acoustic force delivered by ultrasound and resulted in an increased Ca2+ influx into cells. Coupling with a synthetic Ca2+-dependent signaling pathway GVs efficiently enhanced cell stimulation by ultrasound, which expands the potentials of noninvasive sonogenetics cell stimulation.

The Role and Meaning of the Digital Transformation As a Disruptive Innovation on Small and Medium Manufacturing Enterprises.

The research reported in this paper explores the impact of digital transformation as a disruptive innovation on manufacturing SMEs. The research is based on a qualitative Delphi study encompassing 49 experts from eleven EU countries. The paper aims to demonstrate how disruptive innovations affect organizational changes and determine critical factors in organizations that impact the initiating and promoting R&D of disruptive innovation. We discovered that disruptive innovations impact product/process development methods, new production concepts, new materials for products, and new organization plans. Additionally, we identified organizational changes related to the development and use of disruptive innovations in the future. We also indicate how disruptive innovations influence social and technological changes in the organizational environment. The analysis also disclosed three main groups of disruptive innovations and their impact on future smart factory development, namely the following: technological changes, the emergence of innovative products, business models and solutions and organizational culture as one of the crucial key success factors. The analysis also examined the enablers of the successful development/introduction of disruptive innovations, wherein internal and external factors were determined. Additionally, we presented obstacles and the approaches necessary to mitigate them. We can conclude from the findings that in the timeframe of 5-10 years, only the SME that uses/develops disruptive innovations will survive in the market. However, the companies do not always have a clear idea of the meaning of disruptive innovations. Therefore, it is important to set clear goals regarding the achievement of disruptive innovations in companies. It is also necessary to creatively apply presented instruments enabling improvement of organizational changes and apply some additional concepts, which we have suggested.

Engineering and Rewiring of a Calcium-Dependent Signaling Pathway.

An important feature of synthetic biological circuits is their response to physicochemical signals, which enables the external control of cellular processes. Calcium-dependent regulation is an attractive approach for achieving such control, as diverse stimuli induce calcium influx by activating membrane channel receptors. Most calcium-dependent gene circuits use the endogenous nuclear factor of activated T-cells (NFAT) signaling pathway. Here, we employed engineered NFAT transcription factors to induce the potent and robust activation of exogenous gene expression in HEK293T cells. Furthermore, we designed a calcium-dependent transcription factor that does not interfere with NFAT-regulated promoters and potently activates transcription in several mammalian cell types. Additionally, we demonstrate that coupling the circuit to a calcium-selective ion channel resulted in capsaicin- and temperature-controlled gene expression. This engineered calcium-dependent signaling pathway enables tightly controlled regulation of gene expression through different stimuli in mammalian cells and is versatile, adaptable, and useful for a wide range of therapeutic and diagnostic applications.

Psychosocial Risk Factors for Low Back Pain and Absenteeism among Slovenian Professional Drivers.

OBJECTIVE: The aim of this study was to determine the most common psychosocial risk factors for absenteeism and the extent to which low back pain occurs among Slovenian professional drivers as result of various psychosocial risk factors. METHOD: The study involved 275 professional drivers, mostly men (mean age 41.6 years). Statistical data analysis was conducted using SPSS package version 21, MS Excel version 2007 and Pajek, version 3. The main method for data processing was regression analysis. RESULTS: The results of the quantitative survey showed that lower back pain is mostly caused by lifting and carrying heavy loads, inadequate working conditions, poor physical fitness, regular nights out, shift work, and stress. Dissatisfaction with work, shift work and unsuitable working conditions significantly affect the incidence of low back pain. Absenteeism is influenced by factors such as dissatisfaction at work, disrespectful attitude of managers, unsuitable working conditions, personal dissatisfaction, lack of understanding of the partner, and enjoying nightlife on a regular basis. CONCLUSIONS: The study clarifies the unexplained holistic psychosocial risk factors and treatment effects on health in the population of professional drivers. Such factors can lead to absenteeism. The study also provides initial demonstration research in the Slovenian practice. Furthermore, it provides solutions in a holistic approach to solve the problem of risk factors management.

Lower back pain and absenteeism among professional public transport drivers.

Drivers in public transport are subjected to lower back pain. The reason for the pain is associated with the characteristics of the physical position imposed on the worker while performing the job. Lower back pain is the main cause of absenteeism among drivers. The present study includes 145 public transport drivers employed as professional drivers for an average of 14.14 years. Analysis of the data obtained in the study includes the basic descriptive statistics, χ(2) test and multiple regression analysis. Analysis of the incidence of lower back pain showed that the majority of our sample population suffered from pain in the lower back. We found that there are no statistically significant differences between the groups formed by the length of service as a professional driver and incidence of lower back pain; we were also interested in whether or not the risk factors of lower back pain affects the absenteeism of city bus drivers. Analysis of the data has shown that the risk factors of pain in the lower part of the spine do affect the absenteeism of city bus drivers.

Injuries to primary school pupils and secondary school students during physical education classes and in their leisure time.

The study aimed to establish the frequency of injuries in primary and secondary schools during leisure time and physical education classes in school as well as in group and individual sports. The sample included 2842 pupils from nine primary schools and 1235 students from five secondary schools in Slovenia. The data were processed with the SPSS statistical software package and the frequencies and Crosstabs were calculated. The results showed that substantially more pupils and students were injured in their leisure time than during physical education classes. Girls were more frequently injured in group and individual sports practiced during physical education classes and in individual sports practiced in their leisure time, whereas boys suffered more injuries in group sports practiced in their leisure time. As regards group sports, pupils and students were most frequently injured while playing football in their leisure time whereas, during physical education classes, they suffered most injuries in volleyball, followed closely by basketball and football; as regards individual sports, pupils and students were most frequently injured while cycling and rollerblading in their leisure time, whereas during physical education classes they suffered most injuries in athletics.