Engineering of Cell Derived-Nanovesicle as an Alternative to Exosome Therapy.

BACKGROUND: Exosomes, nano-sized vesicles ranging between 30 and 150 nm secreted by human cells, play a pivotal role in long-range intercellular communication and have attracted significant attention in the field of regenerative medicine. Nevertheless, their limited productivity and cost-effectiveness pose challenges for clinical applications. These issues have recently been addressed by cell-derived nanovesicles (CDNs), which are physically synthesized exosome-mimetic nanovesicles from parent cells, as a promising alternative to exosomes. CDNs exhibit structural, physical, and biological properties similar to exosomes, containing intracellular protein and genetic components encapsulated by the cell plasma membrane. These characteristics allow CDNs to be used as regenerative medicine and therapeutics on their own, or as a drug delivery system. METHODS: The paper reviews diverse methods for CDN synthesis, current analysis techniques, and presents engineering strategies to improve lesion targeting efficiency and/or therapeutic efficacy. RESULTS: CDNs, with their properties similar to those of exosomes, offer a cost-effective and highly productive alternative due to their non-living biomaterial nature, nano-size, and readiness for use, allowing them to overcome several limitations of conventional cell therapy methods. CONCLUSION: Ongoing research and enhancement of CDNs engineering, along with comprehensive safety assessments and stability analysis, exhibit vast potential to advance regenerative medicine by enabling the development of efficient therapeutic interventions.

Three-Dimensionally Printed Expandable Structural Electronics Via Multi-Material Printing Room-Temperature-Vulcanizing (RTV) Silicone/Silver Flake Composite and RTV.

Three-dimensional (3D) printing has various applications in many fields, such as soft electronics, robotic systems, biomedical implants, and the recycling of thermoplastic composite materials. Three-dimensional printing, which was only previously available for prototyping, is currently evolving into a technology that can be utilized by integrating various materials into customized structures in a single step. Owing to the aforementioned advantages, multi-functional 3D objects or multi-material-designed 3D patterns can be fabricated. In this study, we designed and fabricated 3D-printed expandable structural electronics in a substrateless auxetic pattern that can be adapted to multi-dimensional deformation. The printability and electrical conductivity of a stretchable conductor (Ag-RTV composite) were optimized by incorporating a lubricant. The Ag-RTV and RTV were printed in the form of conducting voxels and frame voxels through multi-nozzle printing and were arranged in a negative Poisson's ratio pattern with a missing rib structure, to realize an expandable passive component. In addition, the expandable structural electronics were embedded in a soft actuator via one-step printing, confirming the possibility of fabricating stable interconnections in expanding deformation via a missing rib pattern.

Recent advances in wearable iontronic sensors for healthcare applications.

Iontronic sensors have garnered significant attention as wearable sensors due to their exceptional mechanical performance and the ability to maintain electrical performance under various mechanical stimuli. Iontronic sensors can respond to stimuli like mechanical stimuli, humidity, and temperature, which has led to exploration of their potential as versatile sensors. Here, a comprehensive review of the recent researches and developments on several types of iontronic sensors (e.g., pressure, strain, humidity, temperature, and multi-modal sensors), in terms of their sensing principles, constituent materials, and their healthcare-related applications is provided. The strategies for improving the sensing performance and environmental stability of iontronic sensors through various innovative ionic materials and structural designs are reviewed. This review also provides the healthcare applications of iontronic sensors that have gained increased feasibility and broader applicability due to the improved sensing performance. Lastly, outlook section discusses the current challenges and the future direction in terms of the applicability of the iontronic sensors to the healthcare.

Pontin-deficiency causes senescence in fibroblast cells and epidermal keratinocytes but induces apoptosis in cancer cells.

Pontin, a member of the AAA+ ATPase family, plays important roles in a variety of cellular processes, including transcription regulation, DNA damage response, telomerase activity, and cellular transformation. In the previous studies, Pontin deletion in mice was lethal to embryos. Here, we demonstrate that the depletion of Pontin induced cellular senescence in mouse and human fibroblasts as well as in mouse epidermal keratinocytes. Fibroblast cells with Pontin depletion exhibited a defect in cell proliferation without showing apoptosis. Instead, they exhibited senescence-associated phenotypes including increased senescence-associated-ß-galactosidase activity, elevated levels of p16INK4, and senescence-associated secretory phenotypes. Furthermore, conditional deletion of the Pontin gene in epidermal keratinocytes led to abnormal epidermal stratification, which was accompanied by the induction of senescence in Pontin-lacking cells. We found that Pontin depletion induced a spontaneous DNA damage response, which may be a cause of senescence. Contrary to the behavior of normal cells, Pontin depletion in several cancer cells caused apoptotic cell death without exhibiting senescence phenotypes.

Application of Numerical Weather Prediction Data to Estimate Infection Risk of Bacterial Grain Rot of Rice in Korea.

This study was conducted to evaluate usefulness of numerical weather prediction data generated by the Unified Model (UM) for plant disease forecast. Using the UM06- and UM18-predicted weather data, which were released at 0600 and 1800 Universal Time Coordinated (UTC), respectively, by the Korea Meteorological Administration (KMA), disease forecast on bacterial grain rot (BGR) of rice was examined as compared with the model output based on the automated weather stations (AWS)-observed weather data. We analyzed performance of BGRcast based on the UM-predicted and the AWS-observed daily minimum temperature and average relative humidity in 2014 and 2015 from 29 locations representing major rice growing areas in Korea using regression analysis and two-way contingency table analysis. Temporal changes in weather conduciveness at two locations in 2014 were also analyzed with regard to daily weather conduciveness (C i ) and the 20-day and 7-day moving averages of C i for the inoculum build-up phase (C inc ) prior to the panicle emergence of rice plants and the infection phase (C inf ) during the heading stage of rice plants, respectively. Based on C inc and C inf , we were able to obtain the same disease warnings at all locations regardless of the sources of weather data. In conclusion, the numerical weather prediction data from KMA could be reliable to apply as input data for plant disease forecast models. Weather prediction data would facilitate applications of weather-driven disease models for better disease management. Crop growers would have better options for disease control including both protective and curative measures when weather prediction data are used for disease warning.

MBCAST: A Forecast Model for Marssonina Blotch of Apple in Korea.

A disease forecast model for Marssonina blotch of apple was developed based on field observations on airborne spore catches, weather conditions, and disease incidence in 2013 and 2015. The model consisted of the airborne spore model (ASM) and the daily infection rate model (IRM). It was found that more than 80% of airborne spore catches for the experiment period was made during the spore liberation period (SLP), which is the period of days of a rain event plus the following 2 days. Of 13 rain-related weather variables, number of rainy days with rainfall ≥ 0.5 mm per day (L day ), maximum hourly rainfall (P max ) and average daily maximum wind speed (W avg ) during a rain event were most appropriate in describing variations in air-borne spore catches during SLP (S i ) in 2013. The ASM, S i = 30.280+5.860×L day ×P max -2.123×L day ×P max ×W avg was statistically significant and capable of predicting the amount of airborne spore catches during SLP in 2015. Assuming that airborne conidia liberated during SLP cause leaf infections resulting in symptom appearance after 21 days of incubation period, there was highly significant correlation between the estimated amount of airborne spore catches (S i ) and the daily infection rate (R i ). The IRM, R̂ i = 0.039+0.041×S i , was statistically significant but was not able to predict the daily infection rate in 2015. No weather variables showed statistical significance in explaining variations of the daily infection rate in 2013.

Studies on Lytic, Tailed Bacillus cereus-specific Phage for Use in a Ferromagnetoelastic Biosensor as a Novel Recognition Element.

This study investigated the feasibility of the lytic, tailed Bacillus cereus-specific phage for use in a ferromagnetoelastic (FME) biosensor as a novel recognition element. The phage was immobilized at various concentrations through either direct adsorption or a combination of 11-mercapto-1-undecanoic acid (11-MUA) and [N-(3-dimethylaminopropyl)-N'-carbodiimide hydrochloride and N-hydroxysuccinimide (EDC/NHS)]. The effects of time and temperature on its lytic properties were investigated through the exposure of B. cereus (4 and 8 logCFU/ml) to the phage (8 logPFU/ml) for various incubation periods at 22°C and at various temperatures for 30 and 60 min. As the phage concentration increased, both immobilization methods also significantly increased the phage density (p < 0.05). SEM images confirmed that the phage density on the FME platform corresponded to the increased phage concentration. As the combination of 11-MUA and EDC/NHS enhanced the phage density and orientation by up to 4.3-fold, it was selected for use. When various incubation was conducted, no significant differences were observed in the survival rate of B. cereus within 30 min, which was in contrast to the significant decreases observed at 45 and 60 min (p < 0.05). In addition, temperature exerted no significant effects on the survival rate across the entire temperature range. This study demonstrated the feasibility of the lytic, tailed B. cereus-specific phage as a novel recognition element for use in an FME biosensor. Thus, the phage could be placed on the surface of foods for at least 30 min without any significant loss of B. cereus, as a result of the inherent lytic activity of the B. cereus-specific phage as a novel recognition element.

Parafilimonas terrae gen. nov., sp. nov., isolated from greenhouse soil.

A Gram-stain-negative, short rod-shaped, non-flagellated, yellow bacterium, designated strain 5GHs7-2(T), was isolated from a greenhouse soil sample in South Korea. 16S rRNA gene sequence analysis of strain 5GHs7-2(T) indicated that the isolate belonged to the family Chitinophagaceae, and exhibited the highest sequence similarities with members of the genera Terrimonas (89.2-92.6 %), Sediminibacterium (90.8-91.4 %) and Chitinophaga (89.2-91.7 %), Filimonas lacunae YT21(T) (91.7 %), members of the genus Segetibacter (90.2-91.6 %), Parasegetibacter luojiensis RHYL-37(T) (90.9 %) and Flavihumibacter petaseus T41(T) (91.2 %). Flexirubin-type pigments were present. The major cellular fatty acids of the novel strain were iso-C15 : 0, iso-C17 : 0 3-OH and iso-C15 : 1 G. The polar lipid profile consisted of a large amount of phosphatidylethanolamine, and moderate and small amounts of several unknown aminolipids and lipids. The only respiratory quinone of strain 5GHs7-2(T) was MK-7, and the DNA G+C content was 47.6 mol%. On the basis of the evidence presented, it is concluded that strain 5GHs7-2(T) represents a novel species of a new genus in the family Chitinophagaceae, for which the name Parafilimonas terrae gen. nov., sp. nov. is proposed. The type strain of the type species is 5GHs7-2(T) ( = KACC 17343(T) = DSM 28286(T)).

ßTrCP-mediated ubiquitylation regulates protein stability of Mis18ß in a cell cycle-dependent manner.

Ubiquitin E3 ligases including SCF complex are key regulators of cell cycle. Here, we show that Mis18ß, a component of Mis18 complex governing CENP-A localization, is a new substrate of ßTrCP-containing SCF complex. ßTrCP interacted with Mis18ß exclusively during interphase but not during mitosis and mediated proteasomal degradation of Mis18ß leading to the inactivation of Mis18 complex during interphase. In addition, uncontrolled stabilization of Mis18ß caused cell death. Together, we propose that ßTrCP-mediated regulation of Mis18ß stability is a mechanism to restrict centromere function of Mis18 complex from late mitosis to early G1 phase.

Anti-biofilm activities of quercetin and tannic acid against Staphylococcus aureus.

Staphylococcus aureus is a leading cause of nosocomial infections because of its resistance to diverse antibiotics. The formation of a biofilm is one of the mechanisms of drug resistance in S. aureus. The anti-biofilm abilities of 498 plant extracts against S. aureus were examined. Seventy-two plant extracts belonging to 59 genera and 38 families were found to significantly inhibit the formation of biofilms of S. aureus without affecting the growth of planktonic cells. The most active extract, from Alnus japonica, inhibited the formation of biofilms by three S. aureus strains by >70% at 20 µg ml(-1). Transcriptional analyses showed that extract of A. japonica repressed the intercellular adhesion genes icaA and icaD most markedly. Quercetin and tannic acid are major anti-biofilm compounds in the extract of A. japonica. Additionally, the extract of A. japonica and its component compound quercetin, reduced hemolysis by S. aureus. This phenomenon was not observed in the treatment with tannic acid. This study suggests that various plant extracts, such as quercetin and tannic acid, could be used to inhibit the formation of recalcitrant biofilms of S. aureus.

Flavone reduces the production of virulence factors, staphyloxanthin and α-hemolysin, in Staphylococcus aureus.

Staphylococcus aureus is a leading cause of nosocomial infections due to its resistance to diverse antibiotics. This bacterium produces a large number of extracellular virulence factors that are closely associated with specific diseases. In this study, diverse plant flavonoids were investigated to identify a novel anti-virulence compound against two S. aureus strains. Flavone, a backbone compound of flavonoids, at subinhibitory concentration (50 µg/mL), markedly reduced the production of staphyloxanthin and α-hemolysin. This staphyloxanthin reduction rendered the S. aureus cells 100 times more vulnerable to hydrogen peroxide in the presence of flavone. In addition, flavone significantly decreased the hemolysis of human red blood by S. aureus, and the transcriptional level of α-hemolysin gene hla and a global regulator gene sae in S. aureus cells. This finding supported the usefulness of flavone as a potential antivirulence agent against antibiotic-resistant S. aureus.

Acceleration of protease effect on Staphylococcus aureus biofilm dispersal.

Bacterial biofilms are associated with the persistent infections because of their high tolerance to antimicrobial agents. Hence, controlling pathogenic biofilm formation is important in bacteria-related diseases. Staphylococcus aureus is a versatile human pathogen that readily forms biofilms on human tissues and diverse medical devices. As S. aureus can be naturally found in multi-species communities, the supernatants of 28 bacteria were screened to identify new biofilm inhibitory components against S. aureus. The culture supernatant (1%, v/v) of Pseudomonas aeruginosa PAO1 inhibited S. aureus biofilm formation more than 90% without affecting its planktonic cell growth. The P. aeruginosa supernatant contained a high protease activity, which both inhibited S. aureus biofilm formation and detached pre-existing biofilms. An examination of 13 protease-deficient P. aeruginosa mutants identified that LasB elastase is a major antibiofilm protease in P. aeruginosa against S. aureus. Transcriptional analyses showed that P. aeruginosa supernatant induced the expression of endogenous protease genes (aur, clp, scpA, splA, and sspA) and other regulatory genes (agrA, hla, and saeS). Additionally, exogenous proteinase K clearly enhanced the protease activity of S. aureus. Hence, S. aureus accelerated the expression of its own protease genes in the presence of exogenous protease, leading to the rapid dispersal of its biofilm.

Roles of Mis18α in epigenetic regulation of centromeric chromatin and CENP-A loading.

The Mis18 complex has been identified as a critical factor for the centromeric localization of a histone H3 variant, centromeric protein A (CENP-A), which is responsible for the specification of centromere identity in the chromosome. However, the functional role of Mis18 complex is largely unknown. Here, we generated Mis18α conditional knockout mice and found that Mis18α deficiency resulted in lethality at early embryonic stage with severe defects in chromosome segregation caused by mislocalization of CENP-A. Further, we demonstrate Mis18α's crucial role for epigenetic regulation of centromeric chromatin by reinforcing centromeric localization of DNMT3A/3B. Mis18α interacts with DNMT3A/3B, and this interaction is critical for maintaining DNA methylation and hence regulating epigenetic states of centromeric chromatin. Mis18α deficiency led to reduced DNA methylation, altered histone modifications, and uncontrolled noncoding transcripts in centromere region by decreased DNMT3A/3B enrichment. Together, our findings uncover the functional mechanism of Mis18α and its pivotal role in mammalian cell cycle.

Extracellular protease in Actinomycetes culture supernatants inhibits and detaches Staphylococcus aureus biofilm formation.

Bacterial biofilms are associated with chronic infections due to their resistance to antimicrobial agents. Staphylococcus aureus is a versatile human pathogen and can form biofilms on human tissues and diverse medical devices. To identify novel biofilm inhibitors of S. aureus, the supernatants from a library of 458 Actinomycetes strains were screened. The culture supernatants (1% v/v) of more than 10 Actinomycetes strains inhibited S. aureus biofilm formation by more than 80% without affecting the growth. The culture supernatants of these biofilm-reducing Actinomycetes strains contained a protease (equivalent to 0.1 µg proteinase K ml(-1)), which both inhibited S. aureus biofilm formation and detached pre-existing S. aureus biofilms. This study suggests that protease treatment could be a feasible tool to reduce and eradicate S. aureus biofilms.

Low concentrations of honey reduce biofilm formation, quorum sensing, and virulence in Escherichia coli O157:H7.

Bacterial biofilms are associated with persistent infections due to their high resistance to antimicrobial agents. Hence, controlling pathogenic biofilm formation is important in bacteria-related diseases. Honey, at a low concentration of 0.5% (v/v), significantly reduced biofilm formation in enterohemorrhagic Escherichia coli O157:H7 without inhibiting the growth of planktonic cells. Conversely, this concentration did not inhibit commensal E. coli K-12 biofilm formation. Transcriptome analyses showed that honey significantly repressed curli genes (csgBAC), quorum sensing genes (AI-2 importer and indole biosynthesis), and virulence genes (LEE genes). Glucose and fructose in the honeys were found to be key components in reducing biofilm formation by E. coli O157:H7 through the suppression of curli production and AI-2 import. Furthermore, honey, glucose and fructose decreased the colonization of E. coli O157:H7 cells on human HT-29 epithelial cells. These results suggest that low concentrations of honey, such as in honeyed water, can be a practical means for reducing the colonization and virulence of pathogenic E. coli O157:H7.

SUMOylation of RORalpha potentiates transcriptional activation function.

SUMOylation regulates a variety of cellular processes, including control of transcriptional activities of nuclear receptors. Here, we present SUMOylation of orphan nuclear receptor, RORalpha by both SUMO-1 and SUMO-2. SUMOylation of RORalpha occurred on the 240th lysine residue at the hinge region of human protein. PIAS family members, PIASxalpha, PIAS3, and PIASy, increased SUMOylation of RORalpha, whereas SENP2 specifically removed SUMO from RORalpha. SUMOylation-defective mutant form of RORalpha exhibited decreased transcriptional activity on RORalpha-responsive promoters indicating that SUMOylation may positively regulate transcriptional function of RORalpha.