Characterization of the Secretome of a Specific Cell Expressing Mutant Methionyl-tRNA Synthetase in Co-Culture Using Click Chemistry

Co-culture system, in which two or more distinct cell types are cultured together, is advantageous in that it can mimic the environment of the in vivo niche of the cells. In this study, we presented a strategy to analyze the secretome of a specific cell type under the co-culture condition in serum-supplemented media. For the cell-specific secretome analysis, we expressed the mouse mutant methionyl-tRNA synthetase for the incorporation of the non-canonical amino acid, azidonorleucine into the newly synthesized proteins in cells of which the secretome is targeted. The azidonorleucine-tagged secretome could be enriched, based on click chemistry, and distinguished from any other contaminating proteins, either from the cell culture media or the other cells co-cultured with the cells of interest. In order to have more reliable true-positive identifications of cell-specific secretory bodies, we established criteria to exclude any identified human peptide matched to bovine proteins. As a result, we identified a maximum of 719 secreted proteins in the secretome analysis under this co-culture condition. Last, we applied this platform to profile the secretome of mesenchymal stem cells and predicted its therapeutic potential on osteoarthritis based on secretome analysis.     

Functionalization of Tailored Porous Carbon Monolith for Decontamination of Radioactive Substances

As the control over radioactive species becomes critical for the contemporary human life, the development of functional materials for decontamination of radioactive substances has also become important. In this work, a three-dimensional (3D) porous carbon monolith functionalized with Prussian blue particles was prepared through removal of colloidal silica particles from exfoliated graphene/silica composite precursors. The colloidal silica particles with a narrow size distribution were used to act a role of hard template and provide a sufficient surface area that could accommodate potentially hazardous radioactive substances by adsorption. The unique surface and pore structure of the functionalized porous carbon monolith was examined using electron microscopy and energy-dispersive X-ray analysis (EDS). The effective incorporation of PB nanoparticles was confirmed using diverse instrumentations such as X-ray diffraction (XRD), Fourier-transform infrared (FT-IR), and X-ray photoelectron spectroscopy (XPS). A nitrogen adsorption/desorption study showed that surface area and pore volume increased significantly compared with the starting precursor. Adsorption tests were performed with ¹³³Cs ions to examine adsorption isotherms using both Langmuir and Freundlich isotherms. In addition, adsorption kinetics were also investigated and parameters were calculated. The functionalized porous carbon monolith showed a relatively higher adsorption capacity than that of pristine porous carbon monolith and the bulk PB to most radioactive ions such as ¹³³Cs, ⁸⁵Rb, ¹³⁸Ba, ⁸⁸Sr, ¹⁴⁰Ce, and ²⁰⁵Tl. This material can be used for decontamination in expanded application fields.     

Integrity and cell-monolayer permeability of chitosan nanoparticles in simulated gastrointestinal fluids

The objective of this study was to assess the integrity and intestinal permeability of chitosan nanoparticles (CNPs) in simulated gastrointestinal fluids (SGIF). The cell-associated and transported chitosan was quantified and visualized after incubation of the CNPs with Caco-2 cell monolayer with/without SGIF treatment. In order to establish the role of proteins in the SGIF, CNPs were incubated with 4 proteins having different isoelectric points (pI). CNPs incubated with the fluids did not attach to the cell monolayer in contrast to the intact CNPs. Negatively-charged protein formed the complex with CNPs leading to particle size increase, but protected CNPs from disintegration. In contrary, positively-charged protein interacted with cross-linker causing disintegration of CNPs. CNPs incubated with the fluids did not attach to the cell monolayer in contrast to the intact CNPs. The results suggested that the surface charges of CNPs and proteins play a critical role in structural changes of CNPs in biological environment.     

Fabrication of Al matrix composite reinforced with submicrometer-sized Al<Subscript>2</Subscript>O<Subscript>3</Subscript> particles formed by combustion reaction between HEMM Al and V<Subscript>2</Subscript>O<Subscript>5</Subscript> composite particles during sintering

To fabricate an Al-V matrix composite reinforced with submicron-sized Al₂O₃ and Al_xV_y (Al₃V, Al₁₀V) phases, high energy mechanical milling (HEMM) and sintering were employed. By increasing the milling time, the size of mechanically milled powder was significantly reduced. In this study, the average powder size of 59 μm for Al, and 178 μm for V₂O₅ decreased with the formation of a new product, Al-Al₂O₃-Al_xV_y, with a size range from 1.3 μm to 2.6 μm formed by the in-situ combustion reaction during sintering of HEM milled Al and V₂O₅ composite powders. The in-situ reaction between Al and V₂O₅ during the HEMM and sintering transformed the Al₂O₃ and Al_xV_y (Al₃V, Al₁₀V) phases. Most of the reduced V reacted with excess the Al to form Al_xV_y (Al₃V, Al₁₀V) with very little V dissolved into Al matrix. By increasing the milling time and weight percentage of V₂O₅, the hardness of the Al-Al₂O₃-Al_xV_y composite sintered at 1173 K increased. The composite fabricated with the HEMM Al-20wt.%V₂O₅ composite powder and sintering at 1173 K for 2 h had the highest hardness.     

Urban crash-related child pedestrian injury incidence and characteristics associated with injury severity

Objective

Describe age-based urban pedestrian versus auto crash characteristics and identify crash characteristics associated with injury severity.

Materials and methods

Secondary analysis of the 2004–2010 National Highway and Traffic Safety Administration database for Illinois. All persons in Chicago crashes with age data who were listed as pedestrians (n = 7175 child age ≤19 yo, n = 16,398 adult age ≥20 yo) were included. Incidence and crash characteristics were analyzed by age groups and year. Main outcome measures were incidence, crash setting, and injury severity. Multivariate logistic regression analysis was performed to estimate injury severity by crash characteristics.

Results

Overall incidence was higher for child (146.6 per 100,000) versus adult (117.3 per 100,000) pedestrians but case fatality rate was lower (0.7% for children, 1.7% for adults). Child but not adult pedestrian injury incidence declined over time (trend test p < 0.0001 for <5 yo, 5–9 yo, and 10–14 yo; p < 0.05 for 15–19 yo, p = 0.96 for ≥20 yo). Most crashes for both children and adults took place during optimal driving conditions. Injuries were more frequent during warmer months for younger age groups compared to older (χ²p < 0.001). Midblock crashes increased as age decreased (p < 0.0001 for trend). Most crashes occurred at sites with sub-optimal traffic controls but varied by age (p < 0.0001 for trend). Crashes were more likely to be during daylight on dry roads in clear weather conditions for younger age groups compared to older (χ²p < 0.001). Daylight was associated with less severe injury (child OR 0.93, 95% CI 0.87–0.98; adult OR 0.90, 95% CI 0.87–0.93).

Conclusion

The incidence of urban pedestrian crashes declined over time for child subgroups but not for adults. The setting of pedestrian crashes in Chicago today varies by age but is similar to that seen in other urban locales previously. Injuries for all age groups tend to be less severe during daylight conditions. Age-based prevention efforts may prove beneficial.     

Study on the flexoelectric characteristics in the sensing element of a duplex frustum pyramid

Journal of Mechanical Science and Technology - Recently, many researchers have studied to overcome some disadvantages of conventional sensing mechanisms. And the flexoelectric effect draw great...