The ability to generate patient‐specific cells through induced pluripotent stem cell (iPSC) technology has encouraged development of three‐dimensional extracellular matrix (ECM) scaffolds as ...bioactive substrates for cell differentiation with the long‐range goal of bioengineering organs for transplantation. Perfusion decellularization uses the vasculature to remove resident cells, leaving an intact ECM template wherein new cells grow; however, a rigorous evaluative framework assessing ECM structural and biochemical quality is lacking. To address this, we developed histologic scoring systems to quantify fundamental characteristics of decellularized rodent kidneys: ECM structure (tubules, vessels, glomeruli) and cell removal. We also assessed growth factor retention—indicating matrix biofunctionality. These scoring systems evaluated three strategies developed to decellularize kidneys (1% Triton X‐100, 1% Triton X‐100/0.1% sodium dodecyl sulfate (SDS) and 0.02% Trypsin‐0.05% EGTA/1% Triton X‐100). Triton and Triton/SDS preserved renal microarchitecture and retained matrix‐bound basic fibroblast growth factor and vascular endothelial growth factor. Trypsin caused structural deterioration and growth factor loss. Triton/SDS‐decellularized scaffolds maintained 3 h of leak‐free blood flow in a rodent transplantation model and supported repopulation with human iPSC‐derived endothelial cells and tubular epithelial cells ex vivo. Taken together, we identify an optimal Triton/SDS‐based decellularization strategy that produces a biomatrix that may ultimately serve as a rodent model for kidney bioengineering.
The authors validate an optimal detergent‐based protocol for decellularization of rodent whole‐kidney scaffolds, showing that decellularized scaffolds retain an intact vasculature that can be transplanted or re‐endothelialized, wand that the scaffold supports proliferation and tubule formation by human renal cortical tubular epithelial cells.
Automatic processes are usually thought to occur independently of any cognitive resources. This traditional view has been recently challenged by showing that temporal attention to a target stimulus ...is a prerequisite for "automatic" response priming. The event-related potential (ERP) study reported here extends this research by pursuing a somewhat different approach. In two experiments, it was investigated whether masked semantic priming effects can be modulated by temporal attention to the prime using a cueing procedure. We hypothesized that masked priming is amplified when attention is directed to the stimulus stream in the time window of masked prime presentation, even in the absence of any prime awareness. ERPs were recorded while subjects performed a primed lexical decision task. Target words were preceded by semantically related or unrelated masked prime words, which were not consciously identified. A cue stimulus prompted subjects to direct their attention to the stimulus stream either shortly before the masked prime (short cue interval) or a long time interval before. Priming affected the amplitude of the N400 ERP component, an electrophysiological index of semantic processing. Unrelated prime-target pairs elicited a larger N400 than related pairs (N400 priming effect). Most importantly, this masked N400 priming effect was strongest when the cue interval and the stimulus onset asynchronies were short. The present results show that temporal attention to the prime is a prerequisite for obtaining masked N400 priming effects. They also demonstrate that unconscious automatic processes are susceptible to attentional modulation.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Economic Modeling and Inference takes econometrics to a new level by demonstrating how to combine modern economic theory with the latest statistical inference methods to get the most out of economic ...data. This graduate-level textbook draws applications from both microeconomics and macroeconomics, paying special attention to financial and labor economics, with an emphasis throughout on what observations can tell us about stochastic dynamic models of rational optimizing behavior and equilibrium. Bent Jesper Christensen and Nicholas Kiefer show how parameters often thought estimable in applications are not identified even in simple dynamic programming models, and they investigate the roles of extensions, including measurement error, imperfect control, and random utility shocks for inference. When all implications of optimization and equilibrium are imposed in the empirical procedures, the resulting estimation problems are often nonstandard, with the estimators exhibiting nonregular asymptotic behavior such as short-ranked covariance, superconsistency, and non-Gaussianity. Christensen and Kiefer explore these properties in detail, covering areas including job search models of the labor market, asset pricing, option pricing, marketing, and retirement planning. Ideal for researchers and practitioners as well as students, Economic Modeling and Inference uses real-world data to illustrate how to derive the best results using a combination of theory and cutting-edge econometric techniques. * Covers identification and estimation of dynamic programming models * Treats sources of error--measurement error, random utility, and imperfect control * Features financial applications including asset pricing, option pricing, and optimal hedging * Describes labor applications including job search, equilibrium search, and retirement * Illustrates the wide applicability of the approach using micro, macro, and marketing examples
Rhenium catalysts have shown promise to promote carbon neutrality by reducing a prominent greenhouse gas, CO2, to CO and other starting materials. Much research has focused on identifying ...intermediates in the photocatalysis mechanism as well as time scales of relevant ultrafast processes. Recent studies have implemented multidimensional spectroscopies to characterize the catalyst’s ultrafast dynamics as it undergoes the many steps of its photocycle. Two-dimensional infrared (2D-IR) spectroscopy is a powerful method to obtain molecular structure information while extracting time scales of dynamical processes with ultrafast resolution. Many observables result from 2D-IR experiments including vibrational lifetimes, intramolecular redistribution time scales, and, unique to 2D-IR, spectral diffusion, which is highly sensitive to solute–solvent interactions and motional dynamics. Spectral diffusion, a measure of how long a vibrational mode takes to sample its frequency space due to multiple solvent configurations, has various contributing factors. Properties of the solvent, the solute’s structural flexibility, and electronic properties, as well as interactions between the solvent and solute, complicate identifying the origin of the spectral diffusion. With carefully chosen experiments, however, the source of the spectral diffusion can be unveiled. Within the context of a considerable body of previous work, here we discuss the spectral diffusion of several rhenium catalysts at multiple stages in the catalysis. These studies were performed in multiple polar liquids to aid in discovering the contributions of the solvent. We also performed electronic ground state 2D-IR and electronic excited state transient-2D-IR experiments to observe how spectral diffusion changes upon electronic excitation. Our results indicate that with the original Lehn catalyst in THF, relative to the ground state, the spectral diffusion slows by a factor of 3 in the equilibrated triplet metal-to-ligand charge transfer state. We attribute this slowdown to a decrease in dielectric friction as well as an increase in molecular flexibility. It is possible to partially simulate the charge transfer by altering the electron density moderately by adding electron donating or withdrawing substituents symmetrically to the bipyridine ligand. We find that unlike the significant electronic structure change induced by MLCT, such small substituent effects do not influence the spectral diffusion. A solvent study in THF, DMSO, and CH3CN found there to be an explicit solvent dependence that we can correlate to the solvent donicity, which is a measure of its nucleophilicity. Future studies focused on the solvent effects on spectral diffusion in the crucial photoinitiated state can illuminate the role the solvent plays in the catalysis.
In homogeneous photocatalytic reduction of CO
, it is widely assumed that the primary electron transfer from the sacrificial donor to the catalyst is diffusion controlled, thus little attention has ...been paid to optimizing this step. We present spectroscopic evidence that the precursor complex is preformed, driven by preferential solvation, and two-dimensional infrared spectroscopy reveals triethanolamine (donor)/tetrahydrofuran (solvent) exchange in the photocatalyst's solvation shell, reaching greatest magnitude at the known optimal concentration (∼20% v/v TEOA in THF) for catalytically reducing CO
to CO. Transient infrared absorption shows the appearance of the singly reduced catalyst on an ultrafast (<70 ps) time scale, consistent with non-diffusion controlled electron transfer within the preformed precursor complex. Identification of preferential catalyst-cosolvent interactions suggests a revised paradigm for the primary electron transfer, while illuminating the pivotal importance of solvent exchange in determining the overall efficiency of the photocycle.
The CRESST-II experiment uses cryogenic detectors to search for nuclear recoil events induced by the elastic scattering of dark matter particles in CaWO
4
crystals. Given the low energy threshold of ...our detectors in combination with light target nuclei, low mass dark matter particles can be probed with high sensitivity. In this letter we present the results from data of a single detector module corresponding to 52 kg live days. A blind analysis is carried out. With an energy threshold for nuclear recoils of 307 eV we substantially enhance the sensitivity for light dark matter. Thereby, we extend the reach of direct dark matter experiments to the sub- GeV/
c
2
region and demonstrate that the energy threshold is the key parameter in the search for low mass dark matter particles.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
An extensive observational data set, consisting of more than 106 SF6 vertical profiles from MIPAS measurements distributed over the whole globe has been condensed into monthly zonal means of mean age ...of air for the period September 2002 to January 2010, binned at 10° latitude and 1–2 km altitude. The data were analysed with respect to their temporal variation by fitting a regression model consisting of a constant and a linear increase term, 2 proxies for the QBO variation, sinusoidal terms for the seasonal and semi-annual variation and overtones for the correction of the shapes to the observed data set. The impact of subsidence of mesospheric SF6-depleted air and in-mixing into non-polar latitudes on mid-latitudinal absolute age of air and its linear increase was assessed and found to be small. The linear increase of mean age of stratospheric air was found to be positive and partly larger than the trend derived by Engel et al. (2009) for most of the Northern mid-latitudes, the middle stratosphere in the tropics, and parts of the Southern mid-latitudes, as well as for the Southern polar upper stratosphere. Multi-year decrease of age of air was found for the lowermost and the upper stratospheric tropics, for parts of Southern mid-latitudes, and for the Northern polar regions. Analysis of the amplitudes and phases of the seasonal variation shed light on the coupling of stratospheric regions to each other. In particular, the Northern mid-latitude stratosphere is well coupled to the tropics, while the Northern lowermost mid-latitudinal stratosphere is decoupled, confirming the separation of the shallow branch of the Brewer-Dobson circulation from the deep branch. We suggest an overall increased tropical upwelling, together with weakening of mixing barriers, especially in the Northern Hemisphere, as a hypothetical model to explain the observed pattern of linear multi-year increase/decrease, and amplitudes and phase shifts of the seasonal variation.
Inherent and acquired therapeutic resistance in breast cancer remains a major clinical challenge. In human breast cancer samples, overexpression of the oncogenic transcription factor FoxM1 has been ...suggested to be a marker of poor prognosis. In this study, we report that FoxM1 overexpression confers resistance to the human epidermal growth factor receptor 2 monoclonal antibody Herceptin and microtubule-stabilizing drug paclitaxel, both as single agents and in combination. FoxM1 altered microtubule dynamics to protect tumor cells from paclitaxel-induced apoptosis. Mechanistic investigations revealed that the tubulin-destabilizing protein Stathmin, whose expression also confers resistance to paclitaxel, is a direct transcriptional target of FoxM1. Significantly, attenuating FoxM1 expression by small interfering RNA or an alternate reading frame (ARF)-derived peptide inhibitor increased therapeutic sensitivity. Our findings indicate that targeting FoxM1 could relieve therapeutic resistance in breast cancer.
Ultrafast, polarization-selective time-resolved X-ray absorption near-edge structure (XANES) was used to characterize the photochemistry of vitamin B12, cyanocobalamin (CNCbl), in solution. ...Cobalamins are important biological cofactors involved in methyl transfer, radical rearrangement, and light-activated gene regulation, while also holding promise as light-activated agents for spatiotemporal controlled delivery of therapeutics. We introduce polarized femtosecond XANES, combined with UV–visible spectroscopy, to reveal sequential structural evolution of CNCbl in the excited electronic state. Femtosecond polarized XANES provides the crucial structural dynamics link between computed potential energy surfaces and optical transient absorption spectroscopy. Polarization selectivity can be used to uniquely identify electronic contributions and structural changes, even in isotropic samples when well-defined electronic transitions are excited. Our XANES measurements reveal that the structural changes upon photoexcitation occur mainly in the axial direction, where elongation of the axial Co–CN bond and Co–NIm bond on a 110 fs time scale is followed by corrin ring relaxation on a 260 fs time scale. These observations expose features of the potential energy surfaces controlling cobalamin reactivity and deactivation.
During kidney development, canonical Wnt signaling activates differentiation, while the transcription factor Six2 maintains the progenitor pool. These opposing signals help to regulate nephron ...formation and ensure the full complement of nephrons are formed. Since these two factors control differing fates in kidney mesenchyme, we hypothesized that overexpression of Wnt9b in Six2-expressing cells would disrupt kidney formation and may alter cell differentiation decisions in other tissues. We created a transgenic mouse that conditionally expressed the canonical Wnt ligand in the developing kidney, Wnt9b. The transgene is activated by cre recombinase and expresses GFP. We first tested its biological activity using Hoxb7-cre and found that transgenic Wnt9b was capable of inducing differentiation genes and of rescuing kidney development in Wnt9b(-/-) homozygous deficient mice. In contrast, expression of Wnt9b in cells using Six2-cre caused gastrointestinal distress and severe renal failure in adult mice. Transgenic kidneys had numerous cystic tubules and elevated creatinine values (0.652 ± 0.044) compared to wild-type mice (0.119 ± 0.002). These animals also exhibited a malformed pyloric sphincter, duodenogastric reflux, and a transformation of the distal stomach into proximal fate. The gene expression changes observed for the Wnt9b:EGFP transgene were compared to a stabilized β-catenin allele to determine that Wnt9b is activating the canonical Wnt pathway in the tissues analyzed. These results demonstrate that expression of Wnt9b in Six2-positive cells disrupts cell fate decisions in the kidney and the gastrointestinal tract.
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Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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