The basement membrane possesses a rich 3-dimensional nanoscale topography that provides a physical stimulus, which may modulate cell-substratum adhesion. We have investigated the strength of ...cell-substratum adhesion on nanoscale topographic features of a similar scale to that of the native basement membrane. SV40 human corneal epithelial cells were challenged by well-defined fluid shear, and cell detachment was monitored. We created silicon substrata with uniform grooves and ridges having pitch dimensions of 400-4000 nm using X-ray lithography. F-actin labeling of cells that had been incubated for 24 hours revealed that the percentage of aligned and elongated cells on the patterned surfaces was the same regardless of pitch dimension. In contrast, at the highest fluid shear, a biphasic trend in cell adhesion was observed with cells being most adherent to the smaller features. The 400 nm pitch had the highest percentage of adherent cells at the end of the adhesion assay. The effect of substratum topography was lost for the largest features evaluated, the 4000 nm pitch. Qualitative and quantitative analyses of the cells during and after flow indicated that the aligned and elongated cells on the 400 nm pitch were more tightly adhered compared to aligned cells on the larger patterns. Selected experiments with primary cultured human corneal epithelial cells produced similar results to the SV40 human corneal epithelial cells. These findings have relevance to interpretation of cell-biomaterial interactions in tissue engineering and prosthetic design.
Let it grow: The conjugated polymer poly(3,4‐ethylenedioxythiophene) (PEDOT) was synthesized with heparin as the counterion to form a cell culture substrate. The surface of PEDOT:heparin in the ...neutral state associated biologically active growth factors (see picture). Electrochemical in situ oxidation of PEDOT during live cell culture decreased the bioavailability of the growth factor and created an exact onset of neural stem cell differentiation.
This study examined the effect of increased BW on mean retention time (MRT) of both particulate and solute marker, gastrointestinal tract (GIT) development, and fiber digestion in the whole tract of ...growing Saanen goats using the slaughter technique. A total of 58 Saanen goats with initial BW of 15.7 ± 0.9 kg were allocated into 9 treatments with a 3 × 3 factorial arrangement consisting of 3 sexes (female, castrated males, and intact males) and 3 slaughter weights (initial, intermediate, and final; target BW of 16, 23, and 30 kg at slaughter, respectively). They were fed twice daily (0700 and 1600 h) with the identical diets for ad libitum intake. Mean retention time of particulate matter was estimated by in situ determination of indigestible NDF (iNDF), and the MRT of solute marker was determined by Cr-EDTA. Treatment effects were evaluated in a split-plot design, with sex as the main plot and slaughter weight as the subplot. Orthogonal polynomial contrasts were used to determine linear and quadratic effects of slaughter weight, whereas the effect of sex was compared using the Tukey test. The effects of sex and sex × slaughter weight were not significant for most of variables evaluated. The results showed that DMI (% BW) linearly decreased as slaughter weight increased ( < 0.01). Generally wet weight of the total GIT tissues (% BW) decreased and digesta pool sizes (g) linearly increased with increasing slaughter weight ( ≤ 0.05). The ratio of iNDF:NDF for both ingested diet and reticulorumen digesta linearly increased as slaughter weight increased ( ≤ 0.05). The MRT of particles did not change with increasing slaughter weight ( = 0.94). Mean retention time of particulate matter linearly increased in the omasum but linearly decreased in the abomasum with increasing slaughter weight ( < 0.01). Mean retention time of solute marker in the forestomachs linearly increased with increasing slaughter weight ( < 0.01). The results revealed a decreased selectivity with increasing BW, as supported by a greater ratio of iNDF:NDF for ingested diet. Increasing BW led to neither a longer particle MRT in the reticulorumen nor a digestive advantage. The results also indicated that, on average, 91% of fiber digestion occurred in the forestomachs of the goats.
Insulin binds the insulin receptor (IR) and regulates anabolic processes in target tissues. Impaired IR signalling is associated with multiple diseases, including diabetes, cancer and ...neurodegenerative disorders. IRs have been reported to form nanoclusters at the cell membrane in several cell types, even in the absence of insulin binding. Here we exploit the nanoscale spatial organization of the IR to achieve controlled multivalent receptor activation. To control insulin nanoscale spatial organization and valency, we developed rod-like insulin-DNA origami nanostructures carrying different numbers of insulin molecules with defined spacings. Increasing the insulin valency per nanostructure markedly extended the residence time of insulin-DNA origami nanostructures at the receptors. Both insulin valency and spacing affected the levels of IR activation in adipocytes. Moreover, the multivalent insulin design associated with the highest levels of IR activation also induced insulin-mediated transcriptional responses more effectively than the corresponding monovalent insulin nanostructures. In an in vivo zebrafish model of diabetes, treatment with multivalent-but not monovalent-insulin nanostructures elicited a reduction in glucose levels. Our results show that the control of insulin multivalency and spatial organization with nanoscale precision modulates the IR responses, independent of the insulin concentration. Therefore, we propose insulin nanoscale organization as a design parameter in developing new insulin therapies.
Signaling factors including retinoic acid (RA) and thyroid hormone (T3) promote neuronal, oligodendrocyte, and astrocyte differentiation of cortical neural stem cells (NSCs). However, the functional ...specificity of transcriptional repressor checkpoints controlling these differentiation programs remains unclear. Here, we show by genome-wide analysis that histone deacetylase (HDAC)2 and HDAC3 show overlapping and distinct promoter occupancy at neuronal and oligodendrocyte-related genes in NSCs. The absence of HDAC3, but not HDAC2, initiated a neuronal differentiation pathway in NSCs. The ablation of the corepressor NCOR or HDAC2, in conjunction with T3 treatment, resulted in increased expression of oligodendrocyte genes, revealing a direct HDAC2-mediated repression of Sox8 and Sox10 expression. Interestingly, Sox10 was required also for maintaining the more differentiated state by repression of stem cell programming factors such as Sox2 and Sox9. Distinct and nonredundant actions of NCORs and HDACs are thus critical for control of lineage progression and differentiation programs in neural progenitors.
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•ChIP-seq reveals distinct and overlapping occupancy of HDAC2 and HDAC3 in NSCs•Absence of NCOR promotes oligodendrocyte differentiation of NSCs•HDAC2 controls Sox10 expression in OL differentiation via a SOX2-occupied enhancer•Sox10 is required for maintaining the differentiated state in late OL precursors
ChIP-seq in neural stem cells revealed that HDAC2 and HDAC3 show overlapping and distinct promoter occupancy at neuronal and oligodendrocyte-related genes. HDAC3 knockdown initiated neuronal differentiation. Hermanson, Castelo-Branco, and colleagues show that ablation of Ncor or, in the presence of T3, Hdac2 resulted in increased expression of oligodendrocyte genes, unveiling an HDAC2-mediated repression of Sox10 expression. Sox10 was required for maintaining a more differentiated state by repression of stem cell factors including Sox2 and Sox9.
In this work, it is reported a comparison between solid microspheres produced from glass having conventional industrial sand (IS) and glasses made with rice husk ashes (RHA) as a source of silica. ...The same composition was used to fabricate both glasses. The focused was the production of glass microspheres for road horizontal marking, which would attend the type classification called Premix. The microspheres were obtained by the horizontal flame method. The results showed that characteristics such as production yield, morphology, granulometry and, mainly, retroreflectivity measurements are not altered when rice husk ash is present in the glass composition, showing that this can be a sustainable alternative material for this important application.
The aim of this study was to investigate the effects of sex on the net requirements of growth for Ca (NCa), P (NP), Na (NNa), K (NK), and Mg (NMg) in Saanen goats from 5 to 45 kg BW, with or without ...consideration of the degree of maturity. A database containing 209 individual records for Saanen goats (69 castrated males, 71 intact males, and 69 females) was generated from 6 comparative slaughter studies. Total amounts of Ca, P, Na, K, and Mg in the body were fitted to logarithmized allometric equations using empty BW (EBW) or degree of maturity (EBW/mature EBW) as regressors. The equations were fitted using a mixed model, where sex was considered a fixed effect and study was considered a random effect. Net requirements were estimated by the first derivative of the logarithmized allometric equations. Then, a Monte Carlo simulation was used to assess the uncertainty of calculated net requirement values. Without considering the degree of maturity, sex did not affect NCa, NP, and NNa ( > 0.10). Conversely, considering the degree of maturity, NCa and NP of intact males were 5% and 2%, respectively, greater than those of castrated males and females ( < 0.01), and NNa of males (castrated and intact) was 6% greater than that of females ( < 0.01). Regardless of approach used, NCa and NP remained constant, whereas NNa decreased by 32% as BW ranged from 5 to 45 kg. Without considering the degree of maturity, NMg of castrated and intact males were 8% and 18% greater than that of female goats ( = 0.054), respectively. Hereof NMg of castrated and intact males increased by 8% and 15%, respectively, whereas that of females decreased by 8% as BW ranged from 5 to 45 kg. Considering the degree of maturity, NMg of castrated and intact males were 7% and 17% greater than that of female goats ( = 0.054), respectively. In this regard, NMg of castrated and intact males increased 8% and 16%, respectively, whereas that of females decreased by 7% from 5 to 45 kg BW. Both approaches showed that, regardless of sex ( > 0.10), NK decreased by 26% (i.e., without considering the maturity degree) or 27% (i.e., considering the degree of maturity) from 5 to 45 kg BW. Therefore, the consideration of maturity stage highlights differences across sexes in the net macromineral requirements for growth in goats. Elucidation of sex effects on macromineral requirements for growth may be useful for improving the accuracy of recommendations for mineral requirements for dairy goats.
Skeletal muscle plasticity and remodeling are critical for adapting tissue function to use, disuse, and regeneration. The aim of this study was to identify genes and molecular pathways that regulate ...the transition from atrophy to compensatory hypertrophy or recovery from injury. Here, we have used a mouse model of hindlimb unloading and reloading, which causes skeletal muscle atrophy, and compensatory regeneration and hypertrophy, respectively.
We analyzed mouse skeletal muscle at the transition from hindlimb unloading to reloading for changes in transcriptome and extracellular fluid proteome. We then used qRT-PCR, immunohistochemistry, and bulk and single-cell RNA sequencing data to determine Mustn1 gene and protein expression, including changes in gene expression in mouse and human skeletal muscle with different challenges such as exercise and muscle injury. We generated Mustn1-deficient genetic mouse models and characterized them in vivo and ex vivo with regard to muscle function and whole-body metabolism. We isolated smooth muscle cells and functionally characterized them, and performed transcriptomics and proteomics analysis of skeletal muscle and aorta of Mustn1-deficient mice.
We show that Mustn1 (Musculoskeletal embryonic nuclear protein 1, also known as Mustang) is highly expressed in skeletal muscle during the early stages of hindlimb reloading. Mustn1 expression is transiently elevated in mouse and human skeletal muscle in response to intense exercise, resistance exercise, or injury. We find that Mustn1 expression is highest in smooth muscle-rich tissues, followed by skeletal muscle fibers. Muscle from heterozygous Mustn1-deficient mice exhibit differences in gene expression related to extracellular matrix and cell adhesion, compared to wild-type littermates. Mustn1-deficient mice have normal muscle and aorta function and whole-body glucose metabolism. We show that Mustn1 is secreted from smooth muscle cells, and that it is present in arterioles of the muscle microvasculature and in muscle extracellular fluid, particularly during the hindlimb reloading phase. Proteomics analysis of muscle from Mustn1-deficient mice confirms differences in extracellular matrix composition, and female mice display higher collagen content after chemically induced muscle injury compared to wild-type littermates.
We show that, in addition to its previously reported intracellular localization, Mustn1 is a microprotein secreted from smooth muscle cells into the muscle extracellular space. We explore its role in muscle ECM deposition and remodeling in homeostasis and upon muscle injury. The role of Mustn1 in fibrosis and immune infiltration upon muscle injury and dystrophies remains to be investigated, as does its potential for therapeutic interventions.
•Mustn1 expression increases during muscle remodeling.•Mustn1 is expressed in arterioles of the muscle microvasculature.•Mustn1 is secreted by smooth muscle cells.•Loss of Mustn1 changes muscle extracellular matrix composition.
In a deregulated electricity market, market participants define trading strategies and models to assist the decision-making process. Companies whose operation is heavily dependent on this market are ...increasingly adopting electricity price forecasting models to identify sales and purchase contracts with the best price. This paper intends to contribute to the improvement of the decision-making process for purchasing electricity in the Iberian Electricity Market. The purpose is to develop a forecasting model for electricity spot prices based on prices established on the derivatives markets. The model uses Artificial Neural Networks trained with the Extreme Learning Machine algorithm to determine the monthly average spot prices for the next six months and provides a tool for making trading decisions considering the risk of exposure to spot market volatility. The forecasting model was applied in two scenarios: pre-pandemic and pandemic. The results prove that its application can contribute to improving decision-making for trading electricity in the short/medium term. Experimental results considering both scenarios show that the proposed model can provide month-ahead forecasts with an RMSE up to 6.38 €/MWh.
Current feeding systems for goats estimate the energy and protein requirements for pregnancy using data from sheep. The objective of this study was to predict the NE and net protein requirements for ...pregnancy in goats carrying single and twin fetuses and to compare these requirements with those of sheep. Data were compiled from 2 studies with dairy goats and 3 studies with sheep. These studies measured the energy content (EC) and protein content (PC) of the gravid uterus and of the mammary gland using the comparative slaughter technique. The current study was performed as a meta-analysis using an exponential model, comparing species (sheep versus goats) and litter size (single versus twin) from 50 to 140 d of pregnancy. Total EC and total PC in the gravid uterus were similar in goats and sheep carrying a single fetus. Energy and protein contents of the gravid uterus of sheep carrying twins were, on average, 29% greater than that of goats with twins from 80 to 140 d of pregnancy. During pregnancy, EC and PC of the mammary gland in goats carrying singles and twins were, on average, greater than those of sheep by 9 and 24%, respectively, for EC and by 25% for PC for both litter sizes. In conclusion, the gravid uterus and the mammary gland of goats and sheep require different amounts of energy and protein. Sheep carrying twins have the greatest daily NE and net protein requirements for pregnancy followed by goats carrying twins and both species carrying a single fetus. Therefore, it is inappropriate to adopt data from sheep to predict the net pregnancy requirements of goats, and the results found in this study could be relevant to the nutritional management of dairy goats.
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