SNAP29 mediates the assembly of histidine-induced CTP synthase filaments in proximity to the cytokeratin network.

Under metabolic stress, cellular components can assemble into distinct membraneless organelles for adaptation. One such example is cytidine 5'-triphosphate synthase (CTPS, for which there are CTPS1 and CTPS2 forms in mammals), which forms filamentous structures under glutamine deprivation. We have previously demonstrated that histidine (His)-mediated methylation regulates the formation of CTPS filaments to suppress enzymatic activity and preserve the CTPS protein under glutamine deprivation, which promotes cancer cell growth after stress alleviation. However, it remains unclear where and how these enigmatic structures are assembled. Using CTPS-APEX2-mediated in vivo proximity labeling, we found that synaptosome-associated protein 29 (SNAP29) regulates the spatiotemporal filament assembly of CTPS along the cytokeratin network in a keratin 8 (KRT8)-dependent manner. Knockdown of SNAP29 interfered with assembly and relaxed the filament-induced suppression of CTPS enzymatic activity. Furthermore, APEX2 proximity labeling of keratin 18 (KRT18) revealed a spatiotemporal association of SNAP29 with cytokeratin in response to stress. Super-resolution imaging suggests that during CTPS filament formation, SNAP29 interacts with CTPS along the cytokeratin network. This study links the cytokeratin network to the regulation of metabolism by compartmentalization of metabolic enzymes during nutrient deprivation.

Electrically switchable cholesteric gratings based on slit electrodes.

An electrically switchable diffraction grating (ESDG) based on cholesteric liquid crystal (CLC) filled into the cell with slit electrodes is demonstrated in this study. On one hand, with low voltage, the ESDG has high second order diffraction efficiency because of the alternating planar and fingerprint textures. With high voltage, on the other hand, the ESDG has high first order diffraction efficiency because of the alternating planar and homeotropic textures. The first and second order diffraction efficiencies of ESDG are electrically swapped. The maximum diffraction efficiency of the ESDG is approximately 32% at each grating mode.

Electron stream effect in 0.35 Tesla magnetic resonance image guided radiotherapy for breast cancer.

Purpose: This research aimed to analyze electron stream effect (ESE) during magnetic resonance image guided radiotherapy (MRgRT) for breast cancer patients on a MR-Linac (0.35 Tesla, 6MV), with a focus on the prevention of redundant radiation exposure. Materials and methods: RANDO phantom was used with and without the breast attachment in order to represent the patients after breast conserving surgery (BCS) and those received modified radical mastectomy (MRM). The prescription dose is 40.05 Gy in fifteen fractions for whole breast irradiation (WBI) or 20 Gy single shot for partial breast irradiation (PBI). Thirteen different portals of intensity-modulated radiation therapy were created. And then we evaluated dose distribution in five areas (on the skin of the tip of the nose, the chin, the neck, the abdomen and the thyroid.) outside of the irradiated field with and without 0.35 Tesla. In addition, we added a piece of bolus with the thickness of 1cm on the skin in order to compare the ESE difference with and without a bolus. Lastly, we loaded two patients' images for PBI comparison. Results: We found that 0.35 Tesla caused redundant doses to the skin of the chin and the neck as high as 9.79% and 5.59% of the prescription dose in the BCS RANDO model, respectively. For RANDO phantom without the breast accessory (simulating MRM), the maximal dose increase were 8.71% and 4.67% of the prescription dose to the skin of the chin and the neck, respectively. Furthermore, the bolus we added efficiently decrease the unnecessary dose caused by ESE up to 59.8%. Conclusion: We report the first physical investigation on successful avoidance of superfluous doses on a 0.35T MR-Linac for breast cancer patients. Future studies of MRgRT on the individual body shape and its association with ESE influence is warranted.

A novel engineered vascular construct of stem cell-laden 3D-printed PGSA scaffold enhances tissue revascularization.

Development of transplantable engineered tissue has been hampered by lacking vascular network within the engineered tissue. Three-dimensional (3D) printing has emerged as a new technology with great potential in fabrication and customization of geometric microstructure. In this study, utilizing digital light processing system, we manufactured a recently designed novel 3D architecture scaffold with poly(glycerol sebacate) acrylate (PGSA). Vascular construct was subsequently generated by seeding stem cells within this scaffold. PGSA provided inductive substrate in terms of supporting three-germ layer differentiation of embryonic stem cells (ESCs) and also promoting ESCs-derived vascular progenitor cells (VPCs) differentiation into endothelial cells (ECs). Furthermore, the differentiation efficiency of VPCs into ECs on PGSA was much higher than that on collagen IV or fibronectin. The results from seeding VPCs in the rotating hexagonal PGSA scaffold suggest that this architectural framework is highly efficient for cell engraftment in 3D structures. After long-term suspension culture of the VPCs in scaffold under directed EC differentiation condition, VPC-differentiated ECs were populated in the scaffold and expressed EC markers. Transplantation of the vascular construct in mice resulted in formation of new vascular network and integration of the microvasculature within the scaffold into the existing vasculature of host tissue. Importantly, in a mouse model of wound healing, ECs from the transplanted vascular construct directly contributed to revascularization and enhanced blood perfusion at the injured site. Collectively, this transplantable vascular construct provides an innovative alternative therapeutic strategy for vascular tissue engineering.

Microfluidic techniques for enhancing biofuel and biorefinery industry based on microalgae.

This review presents a critical assessment of emerging microfluidic technologies for the application on biological productions of biofuels and other chemicals from microalgae. Comparisons of cell culture designs for the screening of microalgae strains and growth conditions are provided with three categories: mechanical traps, droplets, or microchambers. Emerging technologies for the in situ characterization of microalgae features and metabolites are also presented and evaluated. Biomass and secondary metabolite productivities obtained at microscale are compared with the values obtained at bulk scale to assess the feasibility of optimizing large-scale operations using microfluidic platforms. The recent studies in microsystems for microalgae pretreatment, fractionation and extraction of metabolites are also reviewed. Finally, comments toward future developments (high-pressure/-temperature process; solvent-resistant devices; omics analysis, including genome/epigenome, proteome, and metabolome; biofilm reactors) of microfluidic techniques for microalgae applications are provided.