Scale-up of manufacturing of printed enzyme electrodes for enzymatic power source applications

Production of printable enzymatic power sources was scaled up from laboratory to roll-to-roll (R2R) pilot production. The anode and cathode enzymes were glucose oxidase (GOx) and laccase, respectively. The best laboratory-scale cells had a maximum power and energy density of 1.4 ± 0.1 µW cm^?2 and 5.5 ± 0.2 µWh cm^?2, respectively. These values are 5 and 28 times higher compared to our previously published values. The R2R-produced cells had a maximum power and energy density of 0.40 ± 0.03 µW cm^?2 and 0.6 ± 0.1 µWh cm^?2, respectively. This is 11 % of the best laboratory manufactured cells. It is suspected that the decrease in electrochemical performance originates from the lower mediator amount and higher drying temperature than that of the laboratory produced cells. However, the trials conducted in this work showed that printed enzymatic active layers can be fabricated and dried with a rotary screen-printing machine in R2R process. Hence, fully printed GOx//laccase power sources could be produced from R2R on a large scale for printed electronics applications.     

Lignin and lignin derivatives as components in biobased hot melt adhesives

Hot melt adhesives (HMAs) are formulated for the first time with different lignins as major components, and the developed HMA formulations were tested for gluing paperboard. The best formulations showed equal or even better bond strength compared to a commercial HMA reference. A maximum bond strength of 16.1 N was achieved with a formulation of oxidized cellulose acetate, organosolv lignin, and triethyl citrate, whereas the bond strength of the commercial HMA reference was 10.5 N. The performance was adjusted via the selection of lignin, the formulation, and chemical modification. Lignin modification was not necessary but provided further possibilities for adjusting the properties for different products (reversible vs irreversible adhesive seams) and also for producing plasticizer-free formulations. Modification with tall oil fatty acids enabled the formulation of fully biobased HMAs without any external plasticizer and provided a bond strength as high as high as 8.9 N. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47983.     

Determination of Oxidized Phosphatidylcholines by Hydrophilic Interaction Liquid Chromatography Coupled to Fourier Transform Mass Spectrometry

A novel liquid chromatography-mass spectrometry (LC-MS) approach for analysis of oxidized phosphatidylcholines by an Orbitrap Fourier Transform mass spectrometer in positive electrospray ionization (ESI) coupled to hydrophilic interaction liquid chromatography (HILIC) was developed. This method depends on three selectivity criteria for separation and identification: retention time, exact mass at a resolution of 100,000 and collision induced dissociation (CID) fragment spectra in a linear ion trap. The process of chromatography development showed the best separation properties with a silica-based Kinetex column. This type of chromatography was able to separate all major lipid classes expected in mammalian samples, yielding increased sensitivity of oxidized phosphatidylcholines over reversed phase chromatography. Identification of molecular species was achieved by exact mass on intact molecular ions and CID tandem mass spectra containing characteristic fragments. Due to a lack of commercially available standards, method development was performed with copper induced oxidation products of palmitoyl-arachidonoyl-phosphatidylcholine, which resulted in a plethora of lipid species oxidized at the arachidonoyl moiety. Validation of the method was done with copper oxidized human low-density lipoprotein (LDL) prepared by ultracentrifugation. In these LDL samples we could identify 46 oxidized molecular phosphatidylcholine species out of 99 possible candidates.     

Impact of SGLT2 Inhibitors on Heart Failure: From Pathophysiology to Clinical Effects

Heart failure (HF) affects up to over 20% of patients with type 2 diabetes (T2DM), even more in the elderly. Although, in T2DM, both hyperglycemia and the proinflammatory status induced by insulin resistance are crucial in cardiac function impairment, SGLT2i cardioprotective mechanisms against HF are several. In particular, these beneficial effects seem attributable to the significant reduction of intracellular sodium levels, well-known to exert a cardioprotective role in the prevention of oxidative stress and consequent cardiomyocyte death. From a molecular perspective, patients’ exposure to gliflozins’ treatment mimics nutrient and oxygen deprivation, with consequent autophagy stimulation. This allows to maintain the cellular homeostasis through different degradative pathways. Thus, since their introduction in the clinical practice, the hypotheses on SGLT2i mechanisms of action have changed: from simple glycosuric drugs, with consequent glucose lowering, erythropoiesis enhancing and ketogenesis stimulating, to intracellular sodium-lowering molecules. This provides their consequent cardioprotective effect, which justifies its significant reduction in CV events, especially in populations at higher risk. Finally, the updated clinical evidence of SGLT2i benefits on HF was summarized. Thus, this review aimed to analyze the cardioprotective mechanisms of sodium glucose transporter 2 inhibitors (SGLT2i) in patients with HF, as well as their clinical impact on cardiovascular events.     

The Urokinase Receptor: A Multifunctional Receptor in Cancer Cell Biology. Therapeutic Implications

Proteolysis is a key event in several biological processes; proteolysis must be tightly controlled because its improper activation leads to dramatic consequences. Deregulation of proteolytic activity characterizes many pathological conditions, including cancer. The plasminogen activation (PA) system plays a key role in cancer; it includes the serine-protease urokinase-type plasminogen activator (uPA). uPA binds to a specific cellular receptor (uPAR), which concentrates proteolytic activity at the cell surface, thus supporting cell migration. However, a large body of evidence clearly showed uPAR involvement in the biology of cancer cell independently of the proteolytic activity of its ligand. In this review we will first describe this multifunctional molecule and then we will discuss how uPAR can sustain most of cancer hallmarks, which represent the biological capabilities acquired during the multistep cancer development. Finally, we will illustrate the main data available in the literature on uPAR as a cancer biomarker and a molecular target in anti-cancer therapy.     

Isoindoline-Based Nitroxides as Bioresistant Spin Labels for Protein Labeling through Cysteines and Alkyne-Bearing Noncanonical Amino Acids

Electron paramagnetic resonance (EPR) spectroscopy in combination with site-directed spin labeling (SDSL) is a powerful tool in protein structural research. Nitroxides are highly suitable spin labeling reagents, but suffer from limited stability, particularly in the cellular environment. Herein we present the synthesis of a maleimide- and an azide-modified tetraethyl-shielded isoindoline-based nitroxide (M- and Az-TEIO) for labeling of cysteines or the noncanonical amino acid para-ethynyl-l -phenylalanine (pENF). We demonstrate the high stability of TEIO site-specifically attached to the protein thioredoxin (TRX) against reduction in prokaryotic and eukaryotic environments, and conduct double electron–electron resonance (DEER) measurements. We further generate a rotamer library for the new residue pENF-Az-TEIO that affords a distance distribution that is in agreement with the measured distribution.     

Tilting the Balance: Therapeutic Prospects of CD83 as a Checkpoint Molecule Controlling Resolution of Inflammation

Chronic inflammatory diseases and transplant rejection represent major challenges for modern health care. Thus, identification of immune checkpoints that contribute to resolution of inflammation is key to developing novel therapeutic agents for those conditions. In recent years, the CD83 (cluster of differentiation 83) protein has emerged as an interesting potential candidate for such a “pro-resolution” therapy. This molecule occurs in a membrane-bound and a soluble isoform (mCD83 and sCD83, respectively), both of which are involved in resolution of inflammation. Originally described as a maturation marker on dendritic cells (DCs), mCD83 is also expressed by activated B and T cells as well as regulatory T cells (Tregs) and controls turnover of MHC II molecules in the thymus, and thereby positive selection of CD4⁺ T cells. Additionally, it serves to confine overshooting (auto-)immune responses. Consequently, animals with a conditional deletion of CD83 in DCs or regulatory T cells suffer from impaired resolution of inflammation. Pro-resolving effects of sCD83 became evident in pre-clinical autoimmune and transplantation models, where application of sCD83 reduced disease symptoms and enhanced allograft survival, respectively. Here, we summarize recent advances regarding CD83-mediated resolution of inflammatory responses, its binding partners as well as induced signaling pathways, and emphasize its therapeutic potential for future clinical trials.     

Tuning the phase of circadian entrainment

The circadian clock coordinates daily physiological, metabolic and behavioural rhythms. These endogenous oscillations are synchronized with external cues (‘zeitgebers’), such as daily light and temperature cycles. When the circadian clock is entrained by a zeitgeber, the phase difference ψ between the phase of a clock-controlled rhythm and the phase of the zeitgeber is of fundamental importance for the fitness of the organism. The phase of entrainment ψ depends on the mismatch between the intrinsic period τ and the zeitgeber period T and on the ratio of the zeitgeber strength to oscillator amplitude. Motivated by the intriguing complexity of empirical data and by our own experiments on temperature entrainment of mouse suprachiasmatic nucleus (SCN) slices, we present a theory on how clock and zeitgeber properties determine the phase of entrainment. The wide applicability of the theory is demonstrated using mathematical models of different complexity as well as by experimental data. Predictions of the theory are confirmed by published data on Neurospora crassa strains for different period mismatches τ − T and varying photoperiods. We apply a novel regression technique to analyse entrainment of SCN slices by temperature cycles. We find that mathematical models can explain not only the stable asymptotic phase of entrainment, but also transient phase dynamics. Our theory provides the potential to explore seasonal variations of circadian rhythms, jet lag and shift work in forthcoming studies.     

Improving the Safety and Efficiency of Materials Transfer at a Construction Site by Using an Elevator

The aim of this study was to compare materials transfer at a construction site, either by carrying manually or by using an elevator. Data were collected at a construction site by using two methods: continuous automatic collection of data on the use of the elevator, and measuring and observing the transportation of some products when different delivery systems were used. Health and safety risks were at their highest when transporting products manually at construction sites. Risks decrease when vertical (an elevator) and horizontal (a wheeled device) transportation tools are used. A similar improvement as in health and safety risks can be seen in efficiency results: The working time needed with the elevator was 41% of the purely manual handling time. When wheeled devices were used, the transportation time was decreased to 16%. The early use of a permanent elevator succeeded well and its capacity was large enough. The effective use of the elevator and wheeled devices require that pathways are in better condition than normally.     

Parameter identifiability of nonlinear systems: the role of initial conditions

Identifiability is a fundamental prerequisite for model identification; it concerns uniqueness of the model parameters determined from the input-output data, under ideal conditions of noise-free observations and error-free model structure. In the late 1980s concepts of differential algebra have been introduced in control and system theory. Recently, differential algebra tools have been applied to study the identifiability of dynamic systems described by polynomial equations. These methods all exploit the characteristic set of the differential ideal generated by the polynomials defining the system. In this paper, it will be shown that the identifiability test procedures based on differential algebra may fail for systems which are started at specific initial conditions and that this problem is strictly related to the accessibility of the system from the given initial conditions. In particular, when the system is not accessible from the given initial conditions, the ideal I having as generators the polynomials defining the dynamic system may not correctly describe the manifold of the solution. In this case a new ideal that includes all differential polynomials vanishing at the solution of the dynamic system started from the initial conditions should be calculated. An identifiability test is proposed which works, under certain technical hypothesis, also for systems with specific initial conditions.