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.     

Intratracheal Transplantation of Mesenchymal Stem Cells Attenuates Hyperoxia-Induced Microbial Dysbiosis in the Lungs,Brain, and Gut in Newborn Rats

We attempted to determine whether intratracheal (IT) transplantation of mesenchymal stem cells (MSCs) could simultaneously attenuate hyperoxia-induced lung injuries and microbial dysbiosis of the lungs, brain, and gut in newborn rats. Newborn rats were exposed to hyperoxia (90% oxygen) for 14 days. Human umbilical cord blood-derived MSCs (5 × 10⁵) were transplanted via the IT route on postnatal day (P) five. At P14, the lungs were harvested for histological, biochemical, and microbiome analyses. Bacterial 16S ribosomal RNA genes from the lungs, brain, and large intestine were amplified, pyrosequenced, and analyzed. IT transplantation of MSCs simultaneously attenuated hyperoxia-induced lung inflammation and the ensuing injuries, as well as the dysbiosis of the lungs, brain, and gut. In correlation analyses, lung interleukin-6 (IL-6) levels were significantly positively correlated with the abundance of Proteobacteria in the lungs, brain, and gut, and it was significantly inversely correlated with the abundance of Firmicutes in the gut and lungs and that of Bacteroidetes in the lungs. In conclusion, microbial dysbiosis in the lungs, brain, and gut does not cause but is caused by hyperoxic lung inflammation and ensuing injuries, and IT transplantation of MSCs attenuates dysbiosis in the lungs, brain, and gut, primarily by their anti-oxidative and anti-inflammatory effects.     

Non-Cytokine Protein Profile of the Mesenchymal Stem Cell Secretome That Regulates the Androgen Production Pathway

Polycystic ovary syndrome (PCOS) is the most common endocrine and metabolic disorder in reproductive-aged women, and it typically involves elevated androgen levels. Recently, it has been reported that human bone marrow mesenchymal stem cells (hBM-MSCs) can regulate androgen synthesis pathways. However, the details of the mechanism are still unclear. hBM-MSC-derived secreted factors (the secretome) are promising sources of cell-based therapy as they consist of various types of proteins. It is thus important to know which proteins interact with disease-implicated biomolecules. This work aimed to investigate which secretome components contain the key factor that inhibits testosterone synthesis. In this study, we fractionated hBM-MSC-conditioned media into three fractions based on their molecular weights and found that, of the three fractions, one had the ability to inhibit the androgen-producing genes efficiently. We also analyzed the components of this fraction and established a protein profile of the hBM-MSC secretome, which was shown to inhibit androgen synthesis. Our study describes a set of protein components present in the hBM-MSC secretome that can be used therapeutically to treat PCOS by regulating androgen production for the first time.     

Oxidative Stress and Chemoradiation-Induced Oral Mucositis: A Scoping Review of In Vitro,In Vivo and Clinical Studies

Chemoradiation-induced mucositis is a debilitating condition of the gastrointestinal tract eventuating from antineoplastic treatment. It is believed to occur primarily due to oxidative stress mechanisms, which generate Reactive Oxygen Species (ROS). The aim of this scoping review was to assess the role of oxidative stress in the development of Oral Mucositis (OM). Studies from the literature, published in MEDLINE and SCOPUS, that evaluated the oxidative stress pathways or antioxidant interventions for OM, were retrieved to elucidate the current understanding of their relationship. Studies failing inclusion criteria were excluded, and those suitable underwent data extraction, using a predefined data extraction table. Eighty-nine articles fulfilled criteria, and these were sub-stratified into models of study (in vitro, in vivo, or clinical) for evaluation. Thirty-five clinical studies evaluated antioxidant interventions on OM’s severity, duration, and pain, amongst other attributes. A number of clinical studies sought to elucidate the protective or therapeutic effects of compounds that had been pre-determined to have antioxidant properties, without directly assessing oxidative stress parameters (these were deemed “indirect evidence”). Forty-seven in vivo studies assessed the capacity of various compounds to prevent OM. Findings were mostly consistent, reporting reduced OM severity associated with a reduction in ROS, malondialdehyde (MDA), myeloperoxidase (MPO), but higher glutathione (GSH) and superoxide dismutase (SOD) activity or expression. Twenty-one in vitro studies assessed potential OM therapeutic interventions. The majority demonstrated successful a reduction in ROS, and in select studies, secondary molecules were assessed to identify the mechanism. In summary, this review highlighted numerous oxidative stress pathways involved in OM pathogenesis, which may inform the development of novel therapeutic targets.     

CDDO-Me Attenuates Clasmatodendrosis in CA1 Astrocyte by Inhibiting HSP25-AKT Mediated DRP1-S637 Phosphorylation in Chronic Epilepsy Rats

Clasmatodendrosis is one of the irreversible astroglial degeneration, which is involved in seizure duration and its progression in the epileptic hippocampus. Although sustained heat shock protein 25 (HSP25) induction leads to this autophagic astroglial death, dysregulation of mitochondrial dynamics (aberrant mitochondrial elongation) is also involved in the pathogenesis in clasmatodendrosis. However, the underlying molecular mechanisms of accumulation of elongated mitochondria in clasmatodendritic astrocytes are elusive. In the present study, we found that clasmatodendritic astrocytes showed up-regulations of HSP25 expression, AKT serine (S) 473 and dynamin-related protein 1 (DRP1) S637 phosphorylations in the hippocampus of chronic epilepsy rats. 2-Cyano-3,12-dioxo-oleana-1,9(11)-dien-28-oic acid methyl ester (CDDO-Me; bardoxolone methyl or RTA 402) abrogated abnormal mitochondrial elongation by reducing HSP25 upregulation, AKT S473- and DRP1 S637 phosphorylations. Furthermore, HSP25 siRNA and 3-chloroacetyl-indole (3CAI, an AKT inhibitor) abolished AKT-DRP1-mediated mitochondrial elongation and attenuated clasmatodendrosis in CA1 astrocytes. These findings indicate that HSP25-AKT-mediated DRP1 S637 hyper-phosphorylation may lead to aberrant mitochondrial elongation, which may result in autophagic astroglial degeneration. Therefore, our findings suggest that the dysregulation of HSP25-AKT-DRP1-mediated mitochondrial dynamics may play an important role in clasmatodendrosis, which would have implications for the development of novel therapies against various neurological diseases related to astroglial degeneration.     

User privacy and modern mobile services: are they on the same path?

Perhaps, the most important parameter for any mobile application or service is the way it is delivered and experienced by the end-users, who usually, in due course, decide to keep it on their software portfolio or not. Most would agree that security and privacy have both a crucial role to play toward this goal. In this context, the current paper revolves around a key question: Do modern mobile applications respect the privacy of the end-user? The focus is on the iPhone platform security and especially on user’s data privacy. By the implementation of a DNS poisoning malware and two real attack scenarios on the popular Siri and Tethering services, we demonstrate that the privacy of the end-user is at stake.     

A fault analysis of DC electric arc furnaces with SVC harmonic filters in a mini-mill plant

This paper proposes the most feasible solution to overcome the failure of the 2nd harmonic filter in the static VAR compensator (SVC) which operates with the DC electric arc furnace (EAF) at Gwangyang Steel Mill in Korea. In order to investigate the causes of this failure, various measurements were carried out on the DC EAF and the main transformer at the PCC (point of common connection). It was concluded that the two causes for the failure are; the inrush current in the main transformer, and the parallel resonance between the system impedance and the harmonic filter. Three solutions to suppress the transformer inrush current and another three solutions to avoid the parallel resonance are suggested. The feasibility of these solutions was verified through the computer simulation with PSCAD/EMTDC. The most feasible solution to avoid further failures of the 2nd harmonic filter was selected, based on the estimated result for the six optional solutions in the point of performance and cost.     

Effect of excess PbO on microstructure and orientation of PZT(60/40) films

Lead zirconate titanate [PZT(60/40)] films were deposited by RF-magnetron sputtering using single oxide targets with various levels of excess PbO. The excess PbO in the film played an important role in the pyrochlore-to-perovskite transformation, nucleation and growth processes, orientation control, and crack formation. When 5% or 20% excess PbO was added to the target, pyrochlore phases were created and the films were severely cracked. However, the films had a perovskite structure without any pyrochlore phases when 10% or 15% excess PbO was added to the targets. More interestingly, the crystallographic orientation was strongly dependant on the excess PbO content. A film with a (111) preferred orientation was produced when 10% excess PbO was added to the target. On the other hand, a film with a (100) preferred orientation was deposited by the target with 15% excess PbO. The dielectric, ferroelectric and piezoelectric properties of these films with different orientations and microstructures were examined and correlated with the film structure.     

Supersonic Nozzle Flow Simulations for Particle Coating Applications: Effects of Shockwaves,Nozzle Geometry,Ambient Pressure,and Substrate Location upon Flow Characteristics

Characteristics of supersonic flow are examined with specific regard to nano-particle thin-film coating. Effects of shockwaves, nozzle geometry, chamber pressure, and substrate location were studied computationally. Shockwaves are minimized to reduce fluctuations in flow properties at the discontinuities across diamond shock structures. Nozzle geometry was adjusted to ensure optimal expansion (i.e., P _exit = P _ambient), where shock formation was significantly reduced and flow kinetic energy maximized. When the ambient pressure was reduced from 1 to 0.01316 bar, the nozzle’s diverging angle must be increased to yield the optimum condition of minimized adversed effects. Beyond some critical distance, substrate location did not seem to be a sensitive parameter on flow characteristics when P _amb = 0.01316 bar; however, overly close proximity to the nozzle exit caused flow disturbances inside the nozzle, thereby adversely affecting coating gas flow.