Biosynthesis and spectroscopic characterization of 2-TM fragments encompassing the sequence of a human GPCR, the Y4 receptor

This paper presents a divide‐and‐conquer approach towards obtaining solution structures of G protein‐coupled receptors. The human Y4 receptor was dissected into two to three transmembrane helix fragments, which were individually studied by solution NMR. We systematically compared various biosynthetic routes for the expression of the fragments in Escherichia coli and discuss purification strategies. In particular, we have compared the production of transmembrane (TM) fragments as inclusion bodies by using the ΔTrp leader sequence, with membrane‐directed expression by using Mistic as the fusion partner, and developed methods for enzymatic cleavage. In addition, direct expression of two‐TM fragments into inclusion bodies is a successful route in some cases. With the exception of TM13, we could produce all fragments in isotope‐labeled form in quantities sufficient for NMR studies. Almost complete backbone resonance assignment was obtained for the first two helices, as well as for helices 5 and 7, and a high degree was obtained for TM6, while conformational exchange processes resulted in the disappearance of many signals from TM4. In addition, complete assignments were obtained for all residues of the N‐terminal domain, as well as the extracellular and cytosolic loops (with the exception of an undecapeptide segment in the second extracellular loop, EC2) and for the complete cytosolic C‐terminal tail. In total, backbone resonances of 78 % of all residues were assigned for the Y4 receptor. Predictions of secondary structure based on backbone chemical shifts indicate that most residues from the TM regions adopt helical conformations, with exception of those around polar residues or prolines. However, the domain boundaries differ slightly from those predicted for homology models. We suggest that the obtained chemical shifts might be useful in assigning the full‐length receptor.     

Energy consumption feedback in perspective: integrating Australian data to meta-analyses on in-home displays

Providing homeowners with real-time feedback on their electricity consumption through a dedicated display device has been shown to reduce consumption by approximately 6–10 %. However, recent advances in smart grid technology have enabled larger sample sizes and more representative sample selection and recruitment methods for display trials. By analyzing these factors using data from current studies, this paper argues that a realistic, large-scale conservation effect from feedback is in the range of 3–5 %. Subsequent analysis shows that providing real-time feedback may not be a cost effective strategy for reducing carbon emissions in Australia, but that it may enable additional benefits such as customer retention and peak-load shift.     

Nanoparticle deposition effects on the minimum heat flux point and quench front speed during quenching in water-based alumina nanofluids

The quenching characteristics of metallic rodlets and spheres were investigated in pure water and water-based nanofluids with alumina nanoparticles of 0.1% by volume. The experiments were performed at both saturated and subcooled conditions under atmospheric pressure. The results demonstrate that while the initial quenching behavior in nanofluids is identical to that in pure water, both the minimum heat flux point temperature and quench front speed are significantly enhanced in subsequent quenching repetitions due to nanoparticle deposition. The nanoparticle effects on the quenching process were analyzed with focus on the intermittent liquid–solid contacts in the film boiling regime. It appears that the liquid–solid interaction during such short-lived contacts is more vigorous when a nanoparticle layer with improved surface wettability and roughness is present, which leads to the premature disruption of film boiling and quenching acceleration.     

Functionalized Fe‐Filled Multiwalled Carbon Nanotubes as Multifunctional Scaffolds for Magnetization of Cancer Cells

With the aim to design addressable magnetically‐active carbon nanotubes (CNTs) for cancer treatment, the use of Fe‐filled CNTs (Fe@MWCNTs) as multifunctional scaffolds is reported for exohedrally anchoring a monoclonal antibody (mAb) known to bind a plasma membrane receptor over‐expressed in several cancer cells (EGFR). Comprehensive microscopic (transmission electron microscopy, atomic force microscopy, and scanning electron microscopy) and spectroscopic (Raman, ⁵⁷Fe Mossbauer, energy dispersive spectroscopy, X‐ray photoelectron spectroscopy (XPS), X‐ray diffraction) characterizations reveal the efficient confinement of magnetically‐active Fe phases (α‐Fe and Fe₃C), while compositional evaluations through XPS, thermogravimetric analysis and gel electrophoresis confirm that mAb immobilization onto Fe@MWCNTs occurs. Enzyme‐linked immunosorbent assay (ELISA), confocal microscopy imaging and western blotting confirm the targeting action toward EGFR‐overexpressing cell lines (EGFR+). In vitro magnetic filtration experiments demonstrate that a selective removal of EGFR+ cells from a mixed population of healthy cell lines could be obtained in very short times (≈10 min). Cytotoxicity evaluations by classic cell staining procedures after application of an electromagnetic radiation inducing magnetic fluid hyperthermia (MFH), show a selective suppression of the EGFR+ cell line. Molecular dynamics and docking simulations of the hybrid mAb/Fe@MWCNTs conjugates nicely show how the presence of the CNT framework does not sterically affect the conformational properties of the two antigen binding regions, further supporting the biochemical findings.     

Multiple-project discount rates for cost-benefit analysis in construction projects: a formal risk model for microgeneration renewable energy technologies

The traditional economic approach for appraising the costs and benefits of construction project net present values involves the calculation of net returns for each investment option under different discount rates. An alternative approach consists of multiple-project discount rates based on risk modelling. The example of a portfolio of microgeneration renewable energy technology is presented to demonstrate that risks and future available budget for re-investment can be taken into account when setting discount rates for construction project specifications in the presence of uncertainty. A formal demonstration is carried out through a reversed intertemporal approach of applied general equilibrium. It is demonstrated that risk and the estimated available budget for future re-investment can be included in the simultaneous assessment of the costs and benefits of multiple projects.     

A semiquantitative approach to the evaluation of acute cardiac allograft rejection at endomyocardial biopsy

BACKGROUND: Histopathologic criteria for grading of acute cardiac allograft rejection are focused on the most severe lesion that is recognized among the myocardial fragments provided by each endomyocardial biopsy specimen. Considering the distribution of rejection lesions among all the fragments improved the accuracy in characterizing the severity of rejection in pathologic studies. This study was undertaken to verify the usefulness of a semiquantitative evaluation of endomyocardial biopsy specimens, consisting of the calculation of the proportion of fragments showing rejection in the clinical setting. METHODS: Of the 2386 biopsy specimens obtained during the first posttransplantation year in 168 consecutive cardiac allograft recipients, 290 biopsy specimens constituted by > or = 3 adequate fragments and showing rejection not followed by treatment (n = 159) or being the first biopsy specimen prompting treatment with augmented immunosuppression for that rejection episode (n = 131) were selected. These biopsy specimens (index biopsy specimens) were grouped according to whether rejection was present in < or = 33%, > 33% to < or = 67%, and > 67% of the fragments. The rejection grade (according to the standardized grading system) and the proportion of fragments positive for rejection were correlated with the occurrence of clinical symptoms and signs of rejection at index biopsy and with the results of the next biopsy. RESULTS: Rejections graded > or = 3A were more frequently symptomatic (36% vs 9% for those graded < 3, p < 0.0001), as were those involving increasing proportions of fragments (< or = 33%: 5 of 124, 4%; > 33 to < or = 67%: 13 of 99, 13%; > 67%: 19 of 67, 28% [p < 0.0001]). Spontaneous resolution after untreated biopsies was more frequent in focal (grade 1A and 2) than in diffuse mild (1B) rejections (68% vs 38% [p < 0.04]), whereas progression to grade 3A or greater was less frequent (4% vs 27% [p < 0.01]). Increasing proportions of positive fragments were associated with lower frequencies of spontaneous resolution (p < 0.05) and higher frequencies of worsening (9%, 22%, 43% [p < 0.009]) or progression to grade 3A or greater (2%, 6%, 28% [p < 0.005]). Complete resolution after treatment was less frequent for increasing proportions of positive fragments at index biopsy (80%, 66%, 49% [p < 0.05]). CONCLUSIONS: Diffuse versus focal rejection pattern and the proportion of positive fragments seem to be clinically relevant in terms of occurrence of symptoms, spontaneous evolution, and response to treatment.     

The Effect of Liposomal Curcumin as an Anti-Inflammatory Strategy on Lipopolysaccharide e from Porphyromonas gingivalis Treated Endothelial Committed Neural Crest Derived Stem Cells: Morphological and Molecular Mechanisms

Curcumin, a yellow polyphenol extracted from the turmeric root is used as a diet supplement. It exhibits anti-inflammatory, antioxidant, and antitumor properties by modulating different intracellular mechanisms. Due to their low solubility in water, the curcumin molecules must be encapsulated into liposomes to improve the bioavailability and biomedical potential. For the periodontal tissue and systemic health, it is essential to regulate the local inflammatory response. In this study, the possible beneficial effect of liposomes loaded with curcumin (CurLIP) in neural crest-derived human periodontal ligament stem cells (hPDLSCs) and in endothelial-differentiated hPDLSCs (e-hPDLSCs) induced with an inflammatory stimulus (lipopolysaccharide obtained from Porphyromonas gingivalis, LPS-G) was evaluated. The CurLIP formulation exhibited a significant anti-inflammatory effect by the downregulation of Toll-like receptor-4 (TLR4)/Myeloid differentiation primary response 88 (MyD88)/nuclear factor kappa light chain enhancer of activated B cells (NFkB)/NLR Family Pyrin Domain Containing 3 (NLRP3)/Caspase-1/Interleukin (IL)-1β inflammation cascade and reactive oxygen species (ROS) formation. Moreover, the exposure to LPS-G caused significant alterations in the expression of epigenetic modifiers, such as DNA Methyltransferase 1 (DNMT1) and P300, while the CurLIP treatment showed physiological expression. Overall, our in vitro study provides novel mechanistic insights into the intracellular pathway exert by CurLIP in the regulation of inflammation and epigenetic modifications.     

Experimental methods in chemical engineering: Process simulation

Process simulation software designs equipment, simulates operations, optimizes a plant's configuration (heat exchangers network, for example), estimates operating and capital expenses, and serves as an educational tool. However, mastering the theoretical background minimizes common mistakes such as applying an incorrect thermodynamic method, selecting improper algorithms in the case of tear systems, and setting irrational system specifications. Engineers and researchers will exploit this tool more often in the future as constant advancements in simulation science as well as new models are released continually. Process simulators make it easier to build digital twins and thus will facilitate the implementation of the industry 4.0 guidelines. We highlight the mathematical and technical features of process simulators, as well as the capabilities and the fields of application. A bibliometric map of keywords from articles citing Aspen+, Aspen plus, Hysys, and Pro/II indexed by Web of Science between 2017 and 2020 identified the main research clusters, such as design, optimization, energy or exergy, biomass; H₂ and CO₂ capture, thermodynamics; and separations and techno-economic analysis.     

Zirconium diboride based coatings for thermal protection of re entry vehicles: Effect of MoSi2 addition

The possibility of improving the performances of new generation re-entry vehicles as well as reducing the mission costs is closely connected to the development of new thermal protection systems: innovative heat shield materials, able to withstand higher temperature during the lift re-entry manoeuvre, would permit to improve reusability and easiness of maintenance as well as to increase the manoeuvrability and the payload of vehicles. UHTC materials, and, among them, Zirconium diboride based materials, exhibit outstanding oxidation and thermal shock resistance, high emissivity and very high melting temperature: all these properties make ZrB₂ based materials possible candidates for thermal protection systems of atmospheric re-entry vehicles. The experimental activities described in the present work were focused on ZrB₂ based coatings deposited by plasma spray, containing SiC and MoSi₂ in different ratios: the work aimed at understanding the role of MoSi₂ addition in the oxidation resistance. XRD analysis was carried out on starting powders and plasma sprayed coatings, in order to evaluate the transformations of crystalline phases induced by thermal deposition and to analyse the possible presence of amorphous phase; XPS was also used to study the coatings surface composition and to obtain information about chemical bonds in the materials. Oxidation behaviour was studied carrying out exposure tests up to 1800 °C in static air to compare and model the oxidation kinetics of the different compositions: results show the better behaviour of materials containing MoSi₂. Microstructure of as sprayed and oxidised samples was observed by means of SEM and analysed by EDS to understand the link between dispersion of Mo containing phases and the improved oxidation resistance.