Towards a microfluidics platform for the continuous manufacture of organic and inorganic nanoparticles.

In the last decade, microfluidics has opened new avenues for the synthesis of nanomaterials. However, the adoption of this production technique has been limited to a few high-value, low-production-volume organic nanoparticles. While there are several technical factors that can be attributed to this slow adoption, an important aspect to consider is the lack of a unified platform capable of producing a wide range of nanomaterials. In this work, we highlight a micro-mixing platform that can manufacture both organic and in-organic nanoparticles over a wide size range (nm-µm). We show that the platform can predictably and reproducibly create size and shape-controlled formulations with high homogeneity through input flow parameters. We further explore parallelization of this platform and discuss key technical constraints for high-volume production. We believe that the platform presented in this work can accelerate the adoption of nanomaterials relevant to a range of industries that encompass pharmaceutics, diagnostics, and cosmeceuticals.

The integrin-adhesome is required to maintain muscle structure, mitochondrial ATP production, and movement forces in Caenorhabditis elegans.

The integrin-adhesome network, which contains >150 proteins, is mechano-transducing and located at discreet positions along the cell-cell and cell-extracellular matrix interface. A small subset of the integrin-adhesome is known to maintain normal muscle morphology. However, the importance of the entire adhesome for muscle structure and function is unknown. We used RNA interference to knock down 113 putative Caenorhabditis elegans homologs constituting most of the mammalian adhesome and 48 proteins known to localize to attachment sites in C. elegans muscle. In both cases, we found >90% of components were required for normal muscle mitochondrial structure and/or proteostasis vs. empty vector controls. Approximately half of these, mainly proteins that physically interact with each other, were also required for normal sarcomere and/or adhesome structure. Next we confirmed that the dystrophy observed in adhesome mutants associates with impaired maximal mitochondrial ATP production (P < 0.01), as well as reduced probability distribution of muscle movement forces compared with wild-type animals. Our results show that the integrin-adhesome network as a whole is required for maintaining both muscle structure and function and extend the current understanding of the full complexities of the functional adhesome in vivo.

Efficacy of a novel multi-enzyme feed additive on growth performance, nutrient digestibility, and gut microbiome of weanling pigs fed corn-wheat or wheat-barley-based diet.

One-hundred-and-ninety-two weanling pigs (6.7 kg body weight) were used to evaluate the impact of a carbohydrases-protease enzyme complex (CPEC) on growth performance, nutrient digestibility, and gut microbiome. Pigs were assigned to one of the four dietary treatments for 42 d according to a 2 × 2 factorial arrangement of diet type (low fiber [LF] or high fiber [HF]) and CPEC supplementation (0 or 170 mg/kg diet). The LF diet was prepared as corn-wheat-based diet while the HF diet was wheat-barley-based and contained wheat middlings and canola meal. Each dietary treatment consisted of 12 replicate pens (six replicates per gender) and four pigs per replicate pen. Over the 42-d period, there was no interaction between diet type and CPEC supplementation on growth performance indices of pigs. Dietary addition of CPEC improved (P < 0.05) the body weight of pigs at days 28 and 42 and the gain-to-feed ratio of pigs from days 0 to 14. During the entire experimental period, dietary CPEC supplementation improved (P < 0.05) the average daily gain and gain-to-feed ratio of pigs. There were interactions between diet type and CPEC supplementation on apparent total tract digestibility (ATTD) of dry matter (DM; P < 0.01), gross energy (GE; P < 0.01), and neutral detergent fiber (NDF; P < 0.05) at d 42. Dietary CPEC addition improved (P < 0.05) ATTD of DM, GE, and NDF in the HF diets. At day 43, dietary CPEC addition resulted in improved (P < 0.05) apparent ileal digestibility (AID) of NDF and interactions (P < 0.05) between diet type and CPEC supplementation on AID of DM and crude fiber. Alpha diversity indices including phylogenetic diversity and observed amplicon sequence variants of fecal microbiome increased (P < 0.05) by the addition of CPEC to the HF diets on day 42. An interaction (P < 0.05) between diet type and CPEC addition on Bray-Curtis dissimilarity index and Unweighted UniFrac distances was observed on day 42. In conclusion, CPEC improved weanling pig performance and feed efficiency, especially in wheat-barley diets, while dietary fiber composition had a more significant impact on fecal microbial communities than CPEC administration. The results of this study underscores carbohydrase's potential to boost pig performance without major microbiome changes.

There is a pressing need to enhance livestock production efficiency to meet the growing global demand for meat. Carbohydrases and proteases are enzymes typically added to swine diets to improve nutrient utilization, leading to better growth rates and feed efficiency. This ultimately contributes to sustainable and economically viable pig farming. However, more research is required to better understand how carbohydrases and proteases interact with different diet types to optimize dietary formulations, and how this may influence gut microbiome composition. In this study, 192 weaner pigs (~7 kg) were assigned to a low-fiber diet or a high-fiber diet. Each diet type was with or without a carbohydrases and protease multi-enzyme supplementation. The results showed that adding a multi-enzyme combination to the pigs' diet significantly improved the pig's performance, regardless of diet type. Improvement in nutrient digestibility was more pronounced in pigs fed the high-fiber diet and that dietary fiber had a greater influence on the composition of fecal microbes. In essence, the study demonstrates that the multi-enzyme can boost pig growth and feed efficiency in diets with varying fiber complexity without causing significant changes in their gut microbiome.

Personalizing driver safety interfaces via driver cognitive factors inference.

Recent advances in AI and intelligent vehicle technology hold the promise of revolutionizing mobility and transportation through advanced driver assistance systems (ADAS). Certain cognitive factors, such as impulsivity and inhibitory control have been shown to relate to risky driving behavior and on-road risk-taking. However, existing systems fail to leverage such factors in assistive driving technologies adequately. Varying the levels of these cognitive factors could influence the effectiveness and acceptance of ADAS interfaces. We demonstrate an approach for personalizing driver interaction via driver safety interfaces that are are triggered based on the inference of the driver's latent cognitive states from their driving behavior. To accomplish this, we adopt a data-driven approach and train a recurrent neural network to infer impulsivity and inhibitory control from recent driving behavior. The network is trained on a population of human drivers to infer impulsivity and inhibitory control from recent driving behavior. Using data collected from a high-fidelity vehicle motion simulator experiment, we demonstrate the ability to deduce these factors from driver behavior. We then use these inferred factors to determine instantly whether or not to engage a driver safety interface. This approach was evaluated using leave-one-out cross validation using actual human data. Our evaluations reveal that our personalized driver safety interface that captures the cognitive profile of the driver is more effective in influencing driver behavior in yellow light zones by reducing their inclination to run through them.

Effect of limestone solubility on mineral digestibility and bone ash in nursery pigs fed diets containing graded level of inorganic phosphorus or increasing dose of a novel consensus bacterial 6-phytase variant.

The effect of a novel consensus bacterial 6-phytase variant (PhyG) on total tract digestibility (ATTD) of minerals and bone ash was evaluated in pigs fed diets containing medium- and high-solubility limestone (MSL and HSL, 69.6 and 91.7% solubility, respectively, at 5 min, pH 3.0) in a randomized complete block design. For each limestone, eight diets were formulated: an inorganic phosphate-free negative control (NC) based on wheat, corn, soybean-meal, canola-meal and rice-barn [0.18% standardized total tract digestible (STTD) P and 0.59% Ca]; the NC supplemented with 250, 500, 1,000, or 2,000 FTU/kg of PhyG, and; the NC with added monocalcium phosphate (MCP) and limestone to produce three positive controls (0.33, 0.27, and 0.21% STTD P, and 0.75, 0.70, and 0.64% Ca, respectively; PC1, PC2, PC3). In total, 128 pigs (12.8 ± 1.33 kg, 8 pigs/treatment, housed individually) were adapted for 16 d followed by 4 d of fecal collection. Femurs were collected from euthanized pigs on day 21. Data were analyzed by one-way ANOVA with means separation by Tukey's test, and by factorial analysis (2 x 4: 2 levels of limestone solubility, 4 STTD P levels, and 2 × 5: 2 levels of limestone solubility, 5 PhyG dose levels). Phytase dose-response was analyzed by curve fitting. A consistent negative effect of HSL on ATTD P and Ca was observed in control diets (P < 0.001). Across phytase-supplemented diets, HSL reduced (P < 0.05) ATTD Ca and P (% and g/kg) compared with MSL. Across limestones, increasing phytase dose level increased (P < 0.05) ATTD P exponentially. Limestone solubility had no effect on bone ash, but PhyG linearly increased (P < 0.05) bone ash; 500 FTU/kg or higher maintained bone ash (g/femur) equivalent to PC1. In conclusion, ATTD P and Ca were reduced by a high compared with a medium soluble limestone, but the novel phytase improved ATTD P and Ca independent of limestone solubility.

Microbial phytase is added to commercial pig diets to increase phosphorus (P) availability and reduce P excretion. It is known that an excess of calcium (Ca), mostly sourced from limestone, can affect phytase efficacy. However, less is known about the impact of limestone quality. This study investigated the effect of a medium- compared to a high-soluble limestone (MSL and HSL, respectively), in combination with increasing dose levels of a novel phytase (PhyG), on mineral digestibility and bone mineralization in young pigs. Without phytase, total tract digestibility of P was lower with HSL than MSL, indicating a negative effect of more soluble limestone on mineral digestibility. Increasing the phytase dose increased digestibility of P with either limestone, and reduced the negative effect of HSL at high dose. Bone mineralization was unaffected by limestone but markedly increased by phytase. At 1,000 FTU/kg, PhyG released an estimated 1.89 or 2.32 g/kg of digestible P from monocalcium phosphate in diets containing MSL and HSL, respectively based on bone ash content. The results demonstrate the efficacy of PhyG in young pig diets whilst indicating that limestone solubility can affect phytase efficacy.

Ileal and total tract digestibility of energy and nutrients in pig diets supplemented with a novel consensus bacterial 6-phytase variant.

An experiment was conducted to test the hypothesis that increasing levels of a novel phytase increases the apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of nutrients in diets fed to young pigs. A negative control (NC) diet based on corn, soybean meal, and canola meal that contained approximately 0.83% phytate (i.e., 0.23% phytate-bound P) was formulated to be deficient in Ca, P, and standardized ileal digestible amino acids (AA). Five additional diets were formulated by adding 250, 500, 1,000, 2,000, or 4,000 phytase units/kg of the novel phytase to the NC diets. Eighteen ileal-cannulated pigs (17.81 ± 1.71 kg) were allotted to a 6 × 3 incomplete Latin square design with six diets and three 11-day periods. There were three pigs per diet in each period; therefore, there were nine replicate pigs per diet. The initial 5 d of each period was considered an adaptation period to the diet. For each period, fecal samples were collected via anal stimulation on days 6, 7, 8, and 9, whereas ileal digesta were collected on days 10 and 11 using standard procedures. Results indicated that the AID of crude protein, indispensable AA, and dispensable AA was increased (quadratic, P < 0.05) as the concentration of microbial phytase increased in the diets. Dietary inclusion of the novel phytase at 1,000 or 2,000 FTU/kg increased the AID of total AA from 73.7% to 79.8%. Increasing levels of microbial phytase increased (quadratic, P < 0.05) the AID of dry matter and minerals (i.e., Ca, P, K, Mg, Cu) in the diets. Likewise, a linear increase (P < 0.05) in the AID of ash and Na was observed as the inclusion level of phytase increased in the diets. Increasing levels of microbial phytase increased (linear, P < 0.01) the AID of gross energy (GE) and starch in the diets. A quadratic (P < 0.05) increase in the ATTD of ash, Ca, P, K, and Cu in experimental diets was observed as the concentration of microbial phytase increased in the diets. The ATTD of Mg and GE also increased (linear; P < 0.05) as concentration of dietary phytase increased. In conclusion, the novel microbial phytase used in this experiment was effective in increasing the AID of dry matter, GE, starch, minerals, and AA, as well as the ATTD of gross energy and minerals in diets formulated to be deficient in Ca, P, and AA.

The effect of microbial phytase on amino acid (AA) digestibility has been inconsistent, but in many experiments, relatively low levels of phytase were used and it is not known if greater concentrations of phytase are needed to increase AA digestibility. A novel consensus bacterial 6-phytase variant has been recently developed, but it is not known if this phytase results in increased digestibility of AA and other nutrients. Therefore, an experiment was conducted to test the hypothesis that dietary inclusion of increasing levels of the novel phytase (i.e., 0, 250, 500, 1,000, 2,000, and 4,000 phytase units/kg) increases ileal digestibility of AA and total tract digestibility of energy and minerals in diets for growing pigs. In this experiment, it was demonstrated that increasing levels of phytase increased the apparent ileal digestibility of starch, gross energy, minerals, crude protein, and AA, as well as the apparent total tract digestibility of gross energy and minerals. The impact of phytase on AA digestibility is possibly dependent on diet composition, phytate and phytase sources and concentrations, and pig maturity; however, further research is needed to confirm this.

Influence of a novel consensus bacterial 6-phytase variant on mineral digestibility and bone ash in young growing pigs fed diets with different concentrations of phytate-bound phosphorus.

A 20-d experiment was conducted to test the hypothesis that phytase increases nutrient digestibility, bone ash, and growth performance of pigs fed diets containing 0.23%, 0.29%, or 0.35% phytate-bound P. Within each level of phytate, five diets were formulated to contain 0, 500, 1,000, 2,000, or 4,000 phytase units (FTU)/kg of a novel phytase (PhyG). Three reference diets were formulated by adding a commercial Buttiauxella phytase (PhyB) at 1,000 FTU/kg to diets containing 0.23%, 0.29%, or 0.35% phytate-bound P. A randomized complete block design with 144 individually housed pigs (12.70 ± 4.01 kg), 18 diets, and 8 replicate pigs per diet was used. Pigs were adapted to diets for 15 d followed by 4 d of fecal collection. Femurs were collected on the last day of the experiment. Results indicated that diets containing 0.35% phytate-bound P had reduced (P < 0.01) digestibility of Ca, P, Mg, and K compared with diets containing less phytate-bound P. Due to increased concentration of total P in diets with high phytate, apparent total tract digestible P and bone ash were increased by PhyG to a greater extent in diets with 0.29% or 0.35% phytate-bound P than in diets with 0.23% phytate-bound P (interaction, P < 0.05). At 1,000 FTU/kg, PhyG increased P digestibility and bone P more (P < 0.05) than PhyB. The PhyG increased (P < 0.01) pig growth performance, and pigs fed diets containing 0.35% or 0.29% phytate-bound P performed better (P < 0.01) than pigs fed the 0.23% phytate-bound P diets. In conclusion, the novel phytase (i.e., PhyG) is effective in increasing bone ash, mineral digestibility, and growth performance of pigs regardless of dietary phytate level.

Corrigendum to "Functionality of a next generation biosynthetic bacterial 6-phytase in enhancing phosphorus availability to weaned piglets fed a corn-soybean meal-based diet without added inorganic phosphate" [Animal Nutrition 6/1 (2020) 24-30].

[This corrects the article DOI: 10.1016/j.aninu.2019.11.003.].

Active Intent Disambiguation for Shared Control Robots.

Assistive shared-control robots have the potential to transform the lives of millions of people afflicted with severe motor impairments. The usefulness of shared-control robots typically relies on the underlying autonomy's ability to infer the user's needs and intentions, and the ability to do so unambiguously is often a limiting factor for providing appropriate assistance confidently and accurately. The contributions of this paper are four-fold. First, we introduce the idea of intent disambiguation via control mode selection, and present a mathematical formalism for the same. Second, we develop a control mode selection algorithm which selects the control mode in which the user-initiated motion helps the autonomy to maximally disambiguate user intent. Third, we present a pilot study with eight subjects to evaluate the efficacy of the disambiguation algorithm. Our results suggest that the disambiguation system (a) helps to significantly reduce task effort, as measured by number of button presses, and (b) is of greater utility for more limited control interfaces and more complex tasks. We also observe that (c) subjects demonstrated a wide range of disambiguation request behaviors, with the common thread of concentrating requests early in the execution. As our last contribution, we introduce a novel field-theoretic approach to intent inference inspired by dynamic field theory that works in tandem with the disambiguation scheme.

A Microfluidic Perfusion Platform for In Vitro Analysis of Drug Pharmacokinetic-Pharmacodynamic (PK-PD) Relationships.

Static in vitro cell culture studies cannot capture the dynamic concentration profiles of drugs, nutrients, and other factors that cells experience in physiological systems. This limits the confidence in the translational relevance of in vitro experiments and increases the reliance on empirical testing of exposure-response relationships and dose optimization in animal models during preclinical drug development, introducing additional challenges owing to species-specific differences in drug pharmacokinetics (PK) and pharmacodynamics (PD). Here, we describe the development of a microfluidic cell culture device that enables perfusion of cells under 2D or 3D culture conditions with temporally programmable concentration profiles. Proof-of-concept studies using doxorubicin and gemcitabine demonstrated the ability of the microfluidic PK-PD device to examine dose- and time-dependent effects of doxorubicin as well as schedule-dependent effects of doxorubicin and gemcitabine combination therapy on cell viability using both step-wise drug concentration profiles and species-specific (i.e., mouse, human) drug PK profiles. The results demonstrate the importance of including physiologically relevant dynamic drug exposure profiles during in vitro drug testing to more accurately mimic in vivo drug effects, thereby improving translatability across nonclinical studies and reducing the reliance on animal models during drug development.

Functionality of a next generation biosynthetic bacterial 6-phytase in enhancing phosphorus availability to weaned piglets fed a corn-soybean meal-based diet without added inorganic phosphate.

The utility of a next generation biosynthetic bacterial 6-phytase (PhyG) in restoring bone ash, bone phosphorus (P) content and performance in piglets depleted in P was evaluated. A total of 9 treatments were tested as follows. Treatment 1, a negative control (NC) diet; treatments 2, 3, 4, NC supplemented with 250, 500 or 1,000 FTU/kg of PhyG; treatments 5, 6, NC supplemented with 500 or 1,000 FTU/kg of a commercial Buttiauxella sp phytase (PhyB); treatments 7, 8, 9, NC supplemented with monocalcium phosphate (MCP) to provide 0.7, 1.4 and 1.8 g/kg digestible P, equating to a digestible P content of 1.8, 2.5 and 2.9 g/kg. The latter constituting the positive control (PC) diet with adequate P and calcium (Ca). The NC was formulated without inorganic P (1.1 g digestible P/kg) and reduced in Ca (5.0 g/kg). Additional limestone was added to treatments 7 to 9 to maintain Ca-to-P ratio between 1.2 and 1.3. A total of 162 crossed Pietrain × (Large White × Landrace) 21-d-old piglets (50% males and 50% females) were fed adaptation diets until 42 d old and then assigned to pens with 2 pigs/pen and 9 pens/treatment in a completely randomized block design. Piglets were fed mash diets based on corn and soybean meal ad libitum for 28 d. At the end of the study, one piglet perpen was euthanized and the right feet collected for determination of bone strength, bone ash and mineral content. Compared with the PC, the NC group had reduced average daily gain (ADG) and increased feed conversion ratio (FCR) during all growth phases and overall, and at d 28 (70 d old) NC pigs had bones with reduced ash, Ca and P content (P < 0.05). The PhyG at 250 FTU/kg improved bone ash vs. NC. Increasing PhyG dose linearly or quadratically improved bone ash, ADG and FCR (P < 0.05). At ≥ 500 FTU/kg, both PhyG and PhyB maintained ADG and FCR equivalent to PC. Linear regression analysis was done to compare the measured response parameters to increasing digestible P from MCP. Based on this analysis it was shown that PhyG and PhyB at 1,000 FTU/kg could replace 1.83 and 1.66 g/kg digestible P from MCP in the diet, respectively, on average across metacarpi bone ash, ADG or FCR. These findings suggest that the biosynthetic phytase is highly effective in the tested dietary setting.

Effect of chicken egg anti-F4 antibodies on performance and diarrhea incidences in enterotoxigenic Escherichia coli K88+-challenged piglets.

The aim was to evaluate the effects of dietary supplementation of spay-dried whole egg containing anti-F4 antibodies (SDWE) against recombinantly produced F4 antigens in enterotoxigenic Escherichia coli K88+ (ETEC)-challenged piglets. Twenty-seven 21-d-old and individually housed piglets were randomly allotted to 3 treatments consisting of a wheat-soybean meal basal diet containing either 0 (control egg powder; CEP), 0.1% (SDWE1) or 0.4% (SDWE2) SDWE. After a 7-d adaptation period, blood samples were collected from all pigs, and pigs were weighed and orally challenged with an ETEC inoculum. Blood was sampled at 24 and 48 h post-challenge, and diarrhea incidences and scores were recorded. On d 14, all pigs were weighed and then euthanized to obtain intestinal tissue samples for histomorphology measurement. During the pre-challenge period, pigs fed the SDWE showed a linear improvement (P < 0.05) in average daily gain (ADG) and gain to feed ratio (G:F), but there were no differences among treatments in growth performance during the post-challenge period. Diarrhea incidences and scores, fecal shedding of ETEC, plasma urea nitrogen content and intestinal histomorphology were similar among treatments. The results show that 0.4% SDWE supported greater piglet performance before challenge although such benefits were not evident during the post-challenge period at either 0.1% or 0.4% supplementation.

Combined effects of chitosan and microencapsulated Enterococcus faecalis CG1.0007 probiotic supplementation on performance and diarrhea incidences in enterotoxigenic Escherichia coli K88+ challenged piglets.

The aim of this study was to investigate the combined effects of chitosan oligosaccharide (COS) and a microencapsulated Enterococcus faecalis CG1.0007 probiotic (PRO) on growth performance and diarrhea incidences in enterotoxigenic Escherichia coli (ETEC) K88+ challenged piglets in a 14-d study. Thirty piglets, 7.19 ± 0.52 kg initial BW weaned at 21 ± 1 d, were allotted to 5 treatment groups (n = 6) consisting of a corn-soybean meal diet with no additive (negative control, NC), NC + 0.25% chlortetracycline (positive control, PC), NC + 400 mg/kg COS (COS), NC + 100 mg/kg PRO (PRO) and NC + a combination of COS and PRO (CPRO). Pigs were individually housed in cages, acclimated to treatments for a 7-d period and had ad libitum access to feed and water throughout the study. On d 8, pigs were weighed, blood samples were collected, and then orally challenged with 6 mL (1 × 1011 cfu/mL) of freshly grown ETEC inoculum. During post-challenge period, blood was sampled at 24 and 48 h to determine plasma urea nitrogen (PUN), and diarrhea incidences and fecal consistency scores were recorded from d 9 to 12. On d 14, all pigs were weighed and then euthanized to obtain intestinal tissue samples for histomorphometric measurements. Growth performance responses were similar among treatments during the pre- and post-challenge periods. There were no significant differences in PUN content, incidences of diarrhea, and fecal consistency scores among treatments. The intestinal histomorphology results did not differ significantly among treatments except for PC with increased (P = 0.0001) villus:crypt ratio compared with the NC. Under the conditions of the present study, it can be concluded that supplementation of piglet diets with 400 mg/kg COS, 100 mg/kg microencapsulated PRO or their combination did not significantly improve piglet growth performance both during the pre- and post-ETEC K88+ oral inoculation. Also, there were no significant reduction of incidences and severity of diarrhea after challenge compared with the control group.

Microfluidics-based 3D cell culture models: Utility in novel drug discovery and delivery research.

The implementation of microfluidic devices within life sciences has furthered the possibilities of both academic and industrial applications such as rapid genome sequencing, predictive drug studies, and single cell manipulation. In contrast to the preferred two-dimensional cell-based screening, three-dimensional (3D) systems have more in vivo relevance as well as ability to perform as a predictive tool for the success or failure of a drug screening campaign. 3D cell culture has shown an adaptive response to the recent advancements in microfluidic technologies which has allowed better control over spheroid sizes and subsequent drug screening studies. In this review, we highlight the most significant developments in the field of microfluidic 3D culture over the past half-decade with a special focus on their benefits and challenges down the lane. With the newer technologies emerging, implementation of microfluidic 3D culture systems into the drug discovery pipeline is right around the bend.

Locomotion of C. elegans: a piecewise-harmonic curvature representation of nematode behavior.

Caenorhabditis elegans, a free-living soil nematode, displays a rich variety of body shapes and trajectories during its undulatory locomotion in complex environments. Here we show that the individual body postures and entire trails of C. elegans have a simple analytical description in curvature representation. Our model is based on the assumption that the curvature wave is generated in the head segment of the worm body and propagates backwards. We have found that a simple harmonic function for the curvature can capture multiple worm shapes during the undulatory movement. The worm body trajectories can be well represented in terms of piecewise sinusoidal curvature with abrupt changes in amplitude, wavevector, and phase.