Muscle fiber-type specific expression of UCP3-protein is reported here for the firts time, using immunofluorescence microscopy
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With the alarming rise of antimicrobial resistance, studies on bacteria-surface interactions are both relevant and timely. Scanning electron microscopy and colony forming unit counting are commonly used techniques but require sophisticated sample preparation and long incubation time. Here, we present a direct method based on molecular dynamics simulation of nanostructured surfaces providing in silico predictions, complemented with time-lapse fluorescence imaging to study live interactions of bacteria at the membrane-substrate level. We evaluate its effectiveness in predicting and statistically analyzing the temporal evolution and spatial distribution of prototypical bacteria with costained nucleoids and membranes (E. coli) on surfaces with nanopillars. We observed cell reorientation, clustering, membrane damage, growth inhibition, and in the extreme case of hydrocarbon-coated nanopillars, this was followed by cell disappearance, validating the obtained simulation results. Contrary to commonly used experimental methods, microscopy data are fast processed, in less than 1 h. In particular, the bactericidal effects can be straightforwardly detected and correlated with surface morphology and/or wettability.
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This study provides insights into novel combinations of hydrothermal modifications and mineral enrichment by demonstrating the versatility of this environmentally more benign approach compared to other common chemical starch modifications like crosslinking. Heat-moisture treatment (HMT) (15 % moisture, 100 °C) of native potato starch (NPS) affords granular products that gelatinise at lower temperatures, hold more water as gel, and are more susceptible to enzymatic digestion. Prior mineral enrichment of NPS with sodium, potassium, magnesium and calcium ions yielded significant changes in pasting curves, with monovalent cations increasing peak viscosity, while divalent cations decrease peak viscosity through ionic crosslinking of phosphate groups, allowing further fine tuning of swelling behaviour. Both short and long HMT (4 h and 16 h) triggered partial disruption of crystallinity and an increase in particle size without visible surface damage as evidenced by X-ray diffraction, laser diffraction and scanning electron microscopy. These novel products may find applications where a thickening agent is needed, and high levels of target minerals are desirable like sport nutrition. The viscosity behaviour, available energy and essential minerals may be beneficial to the formulation and nutritional value of energy gels, while adhering to clean-label requirements of today`s food industry.
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Currently the advances in the field of 3D printing are causing a revolution in the (bio-)medical field. With applications ranging from patient-specific anatomical models for surgical preparation to prosthetic limbs and even scaffolds for tissue engineering, the possibilities seem endless. Today, the most widely used method is FDM printing. However, there is still a limited range of biodegradable and biocompatible materials available. Moreover, printed implants like for instance cardiovascular stents require higher resolution than is possible to reach with FDM. High resolution is crucial to avoid e.g. bacterial growth and aid to mechanical strength of the implant. For this reason, it would be interesting to consider stereolithography as alternative to FDM for applications in the (bio-) medical field. Stereolithography uses photopolymerizable resins to make high resolution prints. Because the amount of commercially available resins is limited and hardly biocompatible, here we investigate the possibility of using acrylates and vinylesters in an effort to expand the existing arsenal of biocompatible resins. Mechanical properties are tailorable by varying the crosslink density and by varying the spacer length. To facilitate rapid production of high-resolution prints we use masked SLA (mSLA) as an alternative to conventional SLA. mSLA cures an entire layer at a time and therefore uses less time to complete a print than conventional SLA. Additionally, with mSLA it takes the same time to make 10 prints as it would to make only one. Several formulations were prepared and tested for printability and mechanical strength.
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Light scattering is a fundamental property that can be exploited to create essential devices such as particle analysers. The most common particle size analyser relies on measuring the angle-dependent diffracted light from a sample illuminated by a laser beam. Compared to other non-light-based counterparts, such a laser diffraction scheme offers precision, but it does so at the expense of size, complexity and cost. In this paper, we introduce the concept of a new particle size analyser in a collimated beam configuration using a consumer electronic camera and machine learning. The key novelty is a small form factor angular spatial filter that allows for the collection of light scattered by the particles up to predefined discrete angles. The filter is combined with a light-emitting diode and a complementary metal-oxide-semiconductor image sensor array to acquire angularly resolved scattering images. From these images, a machine learning model predicts the volume median diameter of the particles. To validate the proposed device, glass beads with diameters ranging from 13 to 125 µm were measured in suspension at several concentrations. We were able to correct for multiple scattering effects and predict the particle size with mean absolute percentage errors of 5.09% and 2.5% for the cases without and with concentration as an input parameter, respectively. When only spherical particles were analysed, the former error was significantly reduced (0.72%). Given that it is compact (on the order of ten cm) and built with low-cost consumer electronics, the newly designed particle size analyser has significant potential for use outside a standard laboratory, for example, in online and in-line industrial process monitoring.
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The performance of neural electrodes in physiological fluid, especially in chronic use, is critical for the success of functional electrical stimulation devices. Tips of the Utah electrode arrays (UEAs) were coated with sputtered iridium oxide film (SIROF) and activated iridium oxide film (AIROF) to study the degradation during charge injection consistent with functional electrical stimulation (FES). The arrays were subjected to continuous biphasic, cathodal first, charge balanced (with equal cathodal and anodal pulse widths) current pulses for 7 h (>1 million pulses) at a frequency of 50 Hz. The amplitude and width of the current pulses were varied to determine the damage threshold of the coatings. Degradation was characterized by scanning electron microscopy, inductively coupled plasma mass spectrometry, electrochemical impedance spectroscopy and cyclic voltammetry. The injected charge and charge density per phase were found to play synergistic role in damaging the electrodes. The damage threshold for SIROF coated electrode tips of the UEA was between 60 nC with a charge density of 1.9 mC/cm2 per phase and 80 nC with a charge density of 1.0 mC/cm2 per phase. While for AIROF coated electrode tips, the threshold was between 40 nC with a charge density of 0.9 mC/cm2 per phase and 50 nC with a charge density of 0.5 mC/cm2 per phase. Compared to AIROF, SIROF showed higher damage threshold and therefore is highly recommended to be used as a stimulation material.
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AimsGenetic hypertrophic cardiomyopathy (HCM) is caused by mutations in sarcomere protein-encoding genes (i.e. genotype-positive HCM). In an increasing number of patients, HCM occurs in the absence of a mutation (i.e. genotype-negative HCM). Mitochondrial dysfunction is thought to be a key driver of pathological remodelling in HCM. Reports of mitochondrial respiratory function and specific disease-modifying treatment options in patients with HCM are scarce.Methods and resultsRespirometry was performed on septal myectomy tissue from patients with HCM (n = 59) to evaluate oxidative phosphorylation and fatty acid oxidation. Mitochondrial dysfunction was most notably reflected by impaired NADH-linked respiration. In genotype-negative patients, but not genotype-positive patients, NADH-linked respiration was markedly depressed in patients with an indexed septal thickness ≥10 compared with <10. Mitochondrial dysfunction was not explained by reduced abundance or fragmentation of mitochondria, as evaluated by transmission electron microscopy. Rather, improper organization of mitochondria relative to myofibrils (expressed as a percentage of disorganized mitochondria) was strongly associated with mitochondrial dysfunction. Pre-incubation with the cardiolipin-stabilizing drug elamipretide and raising mitochondrial NAD+ levels both boosted NADH-linked respiration.ConclusionMitochondrial dysfunction is explained by cardiomyocyte architecture disruption and is linked to septal hypertrophy in genotype-negative HCM. Despite severe myocardial remodelling mitochondria were responsive to treatments aimed at restoring respiratory function, eliciting the mitochondria as a drug target to prevent and ameliorate cardiac disease in HCM. Mitochondria-targeting therapy may particularly benefit genotype-negative patients with HCM, given the tight link between mitochondrial impairment and septal thickening in this subpopulation.
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Seven new 1,3,5-cyclohexyltricarboxamide-phenylalanine derivatives were synthesized in order to investigate the effect of the amino acid chirality on the gelating properties of these small molecules in water. Gelation tests have shown that enantiomerically pure homochiral 1,3,5-cyclohexyltricarboxamide-L-phenylalanine is a non-hydrogelator as it crystallizes from water, whereas the heterochiral derivatives with either two L-phenylalanine moieties and one D-phenylalanine (LLD), or vice versa (DDL), are very good hydrogelators. Concentration-dependent gel-to-sol transition-temperature (T(gs)) curves for LLD or DDL gels show a sigmoidal behaviour, which is in contrast to the logarithmic curves generally observed for gels derived from low molecular weight gelators (LMWGs). Such sigmoidal behaviour can be related to interactions between fibre bundles, which give rise to intertwined bundles of fibres. Transmission electron microscopy (TEM) images of LLD and DDL gels show a network of thin, unbranched, fibre bundles with diameters of 20 nm. Right-handed twisted fibre bundles are present in the LLD gel, whereas left-handed structures can be found in the DDL gel. Each bundle of fibres consists of a finite number of primary fibres. Gels consisting of mixtures of gelators, LLD and DDL, and nongelators (LLL or DDD) were investigated by means of T(gs) measurements, CD spectroscopy and TEM. Results show that the incorporation of nongelator molecules into gel fibres occurs; this leads to higher T(gs) values and to changes in the helicity of the fibre bundles. Furthermore, it was found that peripheral functionalization of the homochiral derivatives LLL or DDD by means of a second amino acid or a hydrophilic moiety can overcome the effect of chirality; this process in turn leads to good hydrogelators.
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Om inzicht te krijgen in spierveroudering is genexpressie gemeten in vastus lateralis biopten van jonge en oude mannen en vrouwen. We vonden dat tijdens het ouder worden bij beide geslachten dezelfde categorieën genen in spieren worden aan- en uitgeschakeld (“gereguleerd”); de mate van deze zogenaamde differentiële expressie was echter geslachtsspecifiek. Bij mannen was oxidatieve fosforylering het meest in het oog springende proces, en bij vrouwen was dit celgroei gemedieerd door AKT-signalering. De conclusie is dat dezelfde processen zijn geassocieerd met skeletspierveroudering bij mannen en vrouwen, maar dat de differentiële expressie van die processen geslachtsspecifiek is.
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