•We show how to construct a 2-spanner among polygonal obstacles.•We show how to construct a 6-spanner among polygonal obstacles of degree at most 15.•We show how to construct a 6-spanner among ...polygonal obstacles of degree at most 10.•We show how to construct a 6-spanner among polygonal obstacles of degree at most 7.
Let V be a finite set of vertices in the plane and S be a finite set of polygonal obstacles, where the vertices of S are in V. We show how to construct a plane 2-spanner of the visibility graph of V with respect to S. As this graph can have unbounded degree, we modify it in three easy-to-follow steps, in order to bound the degree to 7 at the cost of slightly increasing the spanning ratio to 6.
Three-dimensional food printing (3DFP) of multicomponent inks fortified with alternative proteins can help drive consumers towards easier and greater acceptance of alternative proteins by familiar ...nature of the final printed products with respects to taste, texture and appearance. Protein fortification is a well-established way of improving the nutritional and functional properties of foods and helping prevent malnutrition. This work focuses on the optimization of multicomponent food inks containing alternative proteins through response surface methodology (RSM) for 3D printing. RSM reduces the number of experiments required to robustly investigate the interrelationships between the desired response and composition of inks, thereby saving precious resources and time. A plant protein, three insect proteins, and an algae protein along with a traditional animal protein were chosen to fortify carrot powder. Usage of carrot as a vegetable food base to which different proteins and xanthan gum were combined served the purpose of adding the flavour and nutrition as well as providing desired rheological properties for printing. The six protein inks were optimised with the addition of hydrocolloid xanthan gum which worked as an excellent shear thinning and hydrating agent. Printability and syneresis were the targeted responses for optimization by RSM with the second order multiple linear regression model. The rheology, microstructure and textural profile of optimised protein inks were also studied. This RSM-based approach for multi-component food ink optimization is envisioned to be widely adopted in 3D food printing.
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•Three-dimensional food printing (3DFP) of food fortified with alternative proteins.•Insect, plant, algal and animal-based food powders utilised for 3DFP covering varied dietary habits.•Response surface methodology used for systematic optimization of multi-component protein inks.•Approach suitable for food ink optimization using responses including printability and syneresis.•Cricket, BSFL and sericin used as insect protein powders.
Benzodiazepines (BZD) and Z-hypnotics are frequently analyzed in forensic laboratories, and in 2012, the designer benzodiazepines (DBZD) emerged on the illegal drug scene. DBZD represent a particular ...challenge demanding new analytical methods. In this work, parallel artificial liquid membrane extraction (PALME) is used for sample preparation of DBZD, BZD, and Z-hypnotics in whole blood prior to UHPLC-MS/MS analysis. PALME of BZD, DBZD, and Z-hypnotics was performed from whole blood samples, and the analytes were extracted across a supported liquid membrane (SLM) and into an acceptor solution of dimethyl sulfoxide and 200 mM formic acid (75:25,
v
/
v
). The method was validated according to EMA guidelines. The method was linear throughout the calibration range (
R
2
> 0.99). Intra- and inter-day accuracy and precision, as well as matrix effects, were within the guideline limit of ± 15%. LOD and LLOQ ranged from 0.10 to 5.0 ng mL
−1
and 3.2 to 160 ng mL
−1
, respectively. Extraction recoveries were reproducible and above 52%. The method was specific, and the analytes were stable in the PALME extracts for 4 and 10 days at 10 and − 20 °C. No carry-over was observed within the calibration range. PALME and UHPLC-MS/MS for the determination of DBZD, BZD, and Z-hypnotics in whole blood are a green and low-cost alternative that provides high sample throughput (96-well format), extensive sample clean-up, good sensitivity, and high reproducibility. The presented method is also the first method incorporating analysis of DBZD, BZD, and Z-hypnotics in whole blood in one efficient analysis.
Graphical abstract
Abstract
Antigen-specific T cells in blood can be detected through the use of soluble MHC-peptide ligands that engage αβTCR. The fluorescent tetramer assay, developed in part by John Altman et al. ...(1) of the Vaccine Research Center, has become a standard tool for immunologists. Evan Newell et al. ( 2) adapted this peptide-MHC tetramer technology to mass cytometry for the purpose of screening of up to 109 different peptide-MHC tetramers in a single human blood sample, as well as analyzing another 23 markers of T-cell phenotype and function using a recombinant form of streptavidin conjugated to metal tags (Maxpar® kits, Fluidigm CA). We will describe the workflow for enumeration and identification of CMV-specific CD8+ T-cells with Neutravidin-metal reagent complexed with HLA-A*0201 CMV pp65 biotinylated monomer (MBL International, MA) simultaneously with metal-labeled surface markers, cisplatin dead-cell identifier, and the use of metal barcoding of several samples into one. There are 198 different biotinylated monomers, commercially available from MBL International, which can be combined with up to 35 isotope-tagged Neutravidin reagents to design a highly multiparametric assay. 1. Altman, J. D. et al. Phenotypic analysis of antigen-specific T lymphocytes. Science 274: 94-96 (1996) 2. Newell, E. W. et al. Combinatorial tetramer staining and mass cytometry analysis facilitate t-cell epitope mapping and characterization. Nat. Biotechnol. 31, 623-629 (2013).
αβ and γδ T cells arise from a common thymocyte progenitor during development in the thymus. Emerging evidence suggests that the pre-T cell receptor (pre-TCR) and γδ T cell receptor (γδTCR) play ...instructional roles in specifying the αβ and γδ T-lineage fates, respectively. Nevertheless, the signaling pathways differentially engaged to specify fate and promote the development of these lineages remain poorly understood. Here, we show that differential activation of the extracellular signal-related kinase (ERK)-early growth response gene (Egr)-inhibitor of DNA binding 3 (Id3) pathway plays a defining role in this process. In particular, Id3 expression served to regulate adoption of the γδ fate. Moreover, Id3 was both necessary and sufficient to enable γδ-lineage cells to differentiate independently of Notch signaling and become competent IFNγ-producing effectors. Taken together, these findings identify Id3 as a central player that controls both adoption of the γδ fate and its maturation in the thymus.
Developing thymocytes bifurcate from a bipotent precursor into αβ- or ...-lineage T cells. Considering this common origin and the fact that the T-cell receptor (TCR) α-, ...-, and ...-chains ...simultaneously rearrange at the double negative (DN) stage of development, the possibility exists that a given DN cell can express and transmit signals through both the pre-TCR and ...-TCR. Here, we tested this scenario by defining the differentiation outcomes and criteria for lineage choice when both TCR-β and ...-TCR are simultaneously expressed in Rag2... DN cells via retroviral transduction. Our results showed that Rag2... DN cells expressing both TCRs developed along the ...-lineage, down-regulated CD24 expression, and up-regulated CD73 expression, showed a ...-biased gene-expression profile, and produced IFN-... in response to stimulation. However, in the absence of Inhibitor of DNA-binding 3 expression and strong ...-TCR ligand, ...-expressing cells showed a lower propensity to differentiate along the ...-lineage. Importantly, differentiation along the ...-lineage was restored by pre-TCR coexpression, which induced greater down-regulation of CD24, higher levels of CD73, Nr4a2, and Rgs1, and recovery of functional competence to produce IFN-... These results confirm a requirement for a strong ...-TCR ligand engagement to promote maturation along the ... T-cell lineage, whereas additional signals from the pre-TCR can serve to enforce a ...-lineage choice in the case of weaker ...-TCR signals. Taken together, these findings further cement the view that the cumulative signal strength sensed by developing DN cells serves to dictate its lineage choice. (ProQuest: ... denotes formulae/symbols omitted.)
‘Circular economy’ and ‘zero waste economy’ are not just 21st century terms but are increasingly becoming part of mainstream narrative to lead an ethical and environmentally conscious life which ...benefits not just humans but the entire ecosystem. Innovative approaches are being targeted to achieve this mission, one being revalorization of food waste to value added products. Food wastage occurs from the initial steps of agricultural produce to industrial manufacturing and processing, to retail and household consumption. The greatest culprit in generating waste is the common households. 3D food printing (3DFP) is one of the emerging technologies that can be effectively utilized to target food wastes from different sources. Consumer acceptance of food waste as edible food still has many barriers. 3DFP can help rescue certain waste types to generate customized and personalized beautiful designs familiar in taste and form for easier acceptability. We have demonstrated the 3D printability of jackfruit seeds that are often underutilized and discarded as waste. Subjecting the waste to simple and effective cooking techniques helped to prepare food ink suitable for 3D printing. Further studies are planned to improve and characterize the inks further to achieve better prints.
‘3D food printing technology is a rapidly advancing technology offering customization in nutrition, shape, taste and texture. Popular foods can be created in exciting and visually appealing forms ...through 3DFP. In this paper, we demonstrate the 3D printability of rice flour mixed with chai poh powder to recreate familiar Asian dish, chwee kueh. 3DFP of yam paste and gingko nut was also conducted for orh nee dish recreation. Extrusion printing was the chosen technique to 3D print different shapes. Two commercially available food printers were used to test these two food ink formulations. Further experimentation is required to improve and characterize the food inks with additional ingredients like sugar, oil to achieve better printability as well as taste and texture’.
A common bipotent thymocyte precursor gives rise to both lineages of T cells, ab and Isup3I. However, the cell intrinsic and extrinsic factors that influence ab- versus Isup3I-lineage bifurcation ...remain controversial. Isup3I T cells play a unique and vital role in host defense, from maintaining integrity at epithelial and mucosal barriers to their newly defined role as an important innate source of interleukin-17. Although a T cell receptor (TCR)-independent fate choice may take place, emerging data supports a model in which the differential signaling capacity of ab and Isup3ITCRs play an instructional role in specifying lineage fate, with strength of signal measured by the amount of ERK/MAPK pathway activation. Here we discuss how the interplay between intrinsic TCR signals and cell extrinsic signals provided by Notch and TCR ligands help to assign and support a final lineage fate decision.
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![Enforcement of [gamma][delt...](http://api.summon.serialssolutions.com/2.0.0/image/issn/XR4PS5YY4K/0027-8424/small "Enforcement of [gamma][delta]-lineage commitment by the pre-T-cell receptor in precursors with weak [gamma][delta]-TCR signals")
