Physicochemical properties of Alaska pollock surimi as affected by salinity and freeze-thaw cycles

The effects of the residual salt of surimi on biochemical and physical properties as affected by various freeze and thaw cycles were examined. Fresh Alaska pollock (Theragra chalcogramma) surimi was mixed with 4.0% sugar, and 5.0% sorbitol, along with eight combinations of salt (0.4, 0.6, 0.8, and 1.0 % NaCl) and sodium polyphosphate (0.25 and 0.5%). Surimi was then vacuum packed and stored at -18°C until used. Freeze-thaw (FT) cycles (0, 3, 6, and 9) were used to mimic long term frozen storage. At the time of gel preparation, each treatment was adjusted to maintain 2% salt and 78% moisture. The pH decreased as the residual salt increased during frozen storage. Salt extractable protein (SEP) and Ca²⁺-ATPase activity decreased as FT cycles extended. Regardless of residual salt and phosphate during frozen storage, whiteness value (L*-3b*) decreased as FT cycles extended. Water retention ability (WRA) and texture significantly decreased at higher salt content (0.8 and 1.0 %) after 9 FT cycles

A high-throughput 3D bioprinted cancer cell migration and invasion model with versatile and broad biological applicability

Understanding the underlying mechanisms of migration and metastasis is a key focus of cancer research. There is an urgent need to develop in vitro 3D tumor models that can mimic physiological cell-cell and cell-extracellular matrix interactions, with high reproducibility and that are suitable for high throughput (HTP) drug screening. Here, we developed a HTP 3D bioprinted migration model using a bespoke drop-on-demand bioprinting platform. This HTP platform coupled with tunable hydrogel systems enables (i) the rapid encapsulation of cancer cells within in vivo tumor mimicking matrices, (ii) in situ and real-time measurement of cell movement, (iii) detailed molecular analysis for the study of mechanisms underlying cell migration and invasion, and (iv) the identification of novel therapeutic options. This work demonstrates that this HTP 3D bioprinted cell migration platform has broad applications across quantitative cell and cancer biology as well as drug screening

A reversible phospho-switch mediated by ULK1 regulates the activity of autophagy protease ATG4B

Upon induction of autophagy, the ubiquitin-like protein LC3 is conjugated to phosphatidylethanolamine (PE) on the inner and outer membrane of autophagosomes to allow cargo selection and autophagosome formation. LC3 undergoes two processing steps, the proteolytic cleavage of pro-LC3 and the de-lipidation of LC3-PE from autophagosomes, both executed by the same cysteine protease ATG4. How ATG4 activity is regulated to co-ordinate these events is currently unknown. Here we find that ULK1, a protein kinase activated at the autophagosome formation site, phosphorylates human ATG4B on serine 316. Phosphorylation at this residue results in inhibition of its catalytic activity in vitro and in vivo. On the other hand, phosphatase PP2A-PP2R3B can remove this inhibitory phosphorylation. We propose that the opposing activities of ULK1-mediated phosphorylation and PP2A-mediated dephosphorylation provide a phospho-switch that regulates the cellular activity of ATG4B to control LC3 processing

Effects of a simulation-based workshop on nursing students' competence in arterial puncture

Objective: To evaluate whether a short simulation-based workshop in radial artery puncture would improve nursing students’ competence to a level in which they could practise the procedure on a live patient without compromising his safety. Methods: Quasi-experimental one-group pretest-posttest study with 111 third-year nursing students. A 1.5-hour simulation-based workshop was implemented. This included a video-lecture, live demonstrations, selfdirected simulated practice in dyads and individual intermittent feedback. Participants’ skills, knowledge and self-efficacy in arterial puncture were measured before and after attending the workshop. Results: After the intervention, a total of 61.1% of the participants showed the level of competence required to safely practice radial artery puncture on a live patient under supervision. Conclusion: Effective simulation-based training in arterial puncture for nursing students does not necessarily need to be resource-intensive. Well-planned, evidence-based training sessions using low-tech simulators could help educators to achieve good educational outcomes and promote patient safety

Histone deacetylase controls adult stem cell aging by balancing the expression of polycomb genes and jumonji domain containing 3

Aging is linked to loss of the self-renewal capacity of adult stem cells. Here, we observed that human multipotent stem cells (MSCs) underwent cellular senescence in vitro. Decreased expression of histone deacetylases (HDACs), followed by downregulation of polycomb group genes (PcGs), such as BMI1, EZH2 and SUZ12, and by upregulation of jumonji domain containing 3 (JMJD3), was observed in senescent MSCs. Similarly, HDAC inhibitors induced cellular senescence through downregulation of PcGs and upregulation of JMJD3. Regulation of PcGs was associated with HDAC inhibitor-induced hypophosphorylation of RB, which causes RB to bind to and decrease the transcriptional activity of E2F. JMJD3 expression regulation was dependant on histone acetylation status at its promoter regions. A histone acetyltransferase (HAT) inhibitor prevented replicative senescence of MSCs. These results suggest that HDAC activity might be important for MSC self-renewal by balancing PcGs and JMJD3 expression, which govern cellular senescence by p16INK4A regulation

Lysyl oxidase drives tumour progression by trapping EGF receptors at the cell surface

Lysyl oxidase (LOX) remodels the tumour microenvironment by cross-linking the extracellular matrix. LOX overexpression is associated with poor cancer outcomes. Here, we find that LOX regulates the epidermal growth factor receptor (EGFR) to drive tumour progression. We show that LOX regulates EGFR by suppressing TGFβ1 signalling through the secreted protease HTRA1. This increases the expression of Matrilin2 (MATN2), an EGF-like domain-containing protein that traps EGFR at the cell surface to facilitate its activation by EGF. We describe a pharmacological inhibitor of LOX, CCT365623, which disrupts EGFR cell surface retention and delays the growth of primary and metastatic tumour cells in vivo. Thus, we show that LOX regulates EGFR cell surface retention to drive tumour progression, and we validate the therapeutic potential of inhibiting this pathway with the small molecule inhibitor CCT365623

Determination of veterinary pharmaceutical runoffs from a swine manure pile using LC-MS/MS

The mass usage of veterinary pharmaceuticals in farms has contributed to environmental pollution in vicinity waters, soils, and sediments from farms and composting facilities. In the present study, we investigated the usage of four antibiotics (viz., lincomycin, sulfamethazine, sulfamethoxazole, and trimethoprim) to understand their contamination routes from livestock manure piles. Residual levels of these antibiotics in a nearby reservoir were set as a positive control (Site 1), and a swine manure pile in a farm (Site 2) and a soil sample around the manure pile (Site 3) were selected for this study. Artificial rainwater was flowed into the manure sample (Site 2), the soil sample around the manure pile (Site 3), and a soil sample around the vicinity river (Site 4). A stream sample (Site 5) around the manure pile and river water near the manure pile (Site 6) were also collected. For qualitative and quantitative analyses, analytical validation was performed, and all the four antibiotics were detected at Site 1 in the concentration range of 0.03-1.6 mu g/L. Lincomycin was the antibiotic with the highest detection level. At Site 2, the detection level of all antibiotics remained at 0.3-17.3 mu g/L, and their residual amounts were continuously detected in subsequent samples with approximately 30-fold decrease. The migration of antibiotics was confirmed to be independent of pH value. Therefore, this study indicates that farm manure pile should be thoroughly managed for antibiotic contamination in vicinity areas with periodical monitoring, especially waterways

Mesenchymal Stem Cells Transfer Mitochondria to the Cells with Virtually No Mitochondrial Function but Not with Pathogenic mtDNA Mutations

It has been reported that human mesenchymal stem cells (MSCs) can transfer mitochondria to the cells with severely compromised mitochondrial function. We tested whether the reported intercellular mitochondrial transfer could be replicated in different types of cells or under different experimental conditions, and tried to elucidate possible mechanism. Using biochemical selection methods, we found exponentially growing cells in restrictive media (uridine− and bromodeoxyuridine [BrdU]+) during the coculture of MSCs (uridine-independent and BrdU-sensitive) and 143B-derived cells with severe mitochondrial dysfunction induced by either long-term ethidium bromide treatment or short-term rhodamine 6G (R6G) treatment (uridine-dependent but BrdU-resistant). The exponentially growing cells had nuclear DNA fingerprint patterns identical to 143B, and a sequence of mitochondrial DNA (mtDNA) identical to the MSCs. Since R6G causes rapid and irreversible damage to mitochondria without the removal of mtDNA, the mitochondrial function appears to be restored through a direct transfer of mitochondria rather than mtDNA alone. Conditioned media, which were prepared by treating mtDNA-less 143B ρ0 cells under uridine-free condition, induced increased chemotaxis in MSC, which was also supported by transcriptome analysis. Cytochalasin B, an inhibitor of chemotaxis and cytoskeletal assembly, blocked mitochondrial transfer phenomenon in the above condition. However, we could not find any evidence of mitochondrial transfer to the cells harboring human pathogenic mtDNA mutations (A3243G mutation or 4,977 bp deletion). Thus, the mitochondrial transfer is limited to the condition of a near total absence of mitochondrial function. Elucidation of the mechanism of mitochondrial transfer will help us create a potential cell therapy-based mitochondrial restoration or mitochondrial gene therapy for human diseases caused by mitochondrial dysfunction

Observation of associated near-side and away-side long-range correlations in √sNN=5.02 TeV proton-lead collisions with the ATLAS detector

Two-particle correlations in relative azimuthal angle (Δϕ) and pseudorapidity (Δη) are measured in √sNN=5.02  TeV p+Pb collisions using the ATLAS detector at the LHC. The measurements are performed using approximately 1  μb-1 of data as a function of transverse momentum (pT) and the transverse energy (ΣETPb) summed over 3.1<η<4.9 in the direction of the Pb beam. The correlation function, constructed from charged particles, exhibits a long-range (2<|Δη|<5) “near-side” (Δϕ∼0) correlation that grows rapidly with increasing ΣETPb. A long-range “away-side” (Δϕ∼π) correlation, obtained by subtracting the expected contributions from recoiling dijets and other sources estimated using events with small ΣETPb, is found to match the near-side correlation in magnitude, shape (in Δη and Δϕ) and ΣETPb dependence. The resultant Δϕ correlation is approximately symmetric about π/2, and is consistent with a dominant cos⁡2Δϕ modulation for all ΣETPb ranges and particle pT