Effect of Cell Age and Membrane Rigidity on Red Blood Cell Shape in Capillary Flow

Blood flow in the microcirculatory system is crucially affected by intrinsic red blood cell (RBC) properties, such as their deformability. In the smallest vessels of this network, RBCs adapt their shapes to the flow conditions. Although it is known that the age of RBCs modifies their physical properties, such as increased cytosol viscosity and altered viscoelastic membrane properties, the evolution of their shape-adapting abilities during senescence remains unclear. In this study, we investigated the effect of RBC properties on the microcapillary in vitro flow behavior and their characteristic shapes in microfluidic channels. For this, we fractioned RBCs from healthy donors according to their age. Moreover, the membranes of fresh RBCs were chemically rigidified using diamide to study the effect of isolated graded-membrane rigidity. Our results show that a fraction of stable, asymmetric, off-centered slipper-like cells at high velocities decreases with increasing age or diamide concentration. However, while old cells form an enhanced number of stable symmetric croissants at the channel centerline, this shape class is suppressed for purely rigidified cells with diamide. Our study provides further knowledge about the distinct effects of age-related changes of intrinsic cell properties on the single-cell flow behavior of RBCs in confined flows due to inter-cellular age-related cell heterogeneity

The Cornell Net Carbohydrate and Protein System:Updates to the model and evaluation of version 6.5

AbstractNew laboratory and animal sampling methods and data have been generated over the last 10 yr that had the potential to improve the predictions for energy, protein, and AA supply and requirements in the Cornell Net Carbohydrate and Protein System (CNCPS). The objectives of this study were to describe updates to the CNCPS and evaluate model performance against both literature and on-farm data. The changes to the feed library were significant and are reported in a separate manuscript. Degradation rates of protein and carbohydrate fractions were adjusted according to new fractionation schemes, and corresponding changes to equations used to calculate rumen outflows and postrumen digestion were presented. In response to the feed-library changes and an increased supply of essential AA because of updated contents of AA, a combined efficiency of use was adopted in place of separate calculations for maintenance and lactation to better represent the biology of the cow. Four different data sets were developed to evaluate Lys and Met requirements, rumen N balance, and milk yield predictions. In total 99 peer-reviewed studies with 389 treatments and 15 regional farms with 50 different diets were included. The broken-line model with plateau was used to identify the concentration of Lys and Met that maximizes milk protein yield and content. Results suggested concentrations of 7.00 and 2.60% of metabolizable protein (MP) for Lys and Met, respectively, for maximal protein yield and 6.77 and 2.85% of MP for Lys and Met, respectively, for maximal protein content. Updated AA concentrations were numerically higher for Lys and 11 to 18% higher for Met compared with CNCPS v6.0, and this is attributed to the increased content of Met and Lys in feeds that were previously incorrectly analyzed and described. The prediction of postruminal flows of N and milk yield were evaluated using the correlation coefficient from the BLUP (R2BLUP) procedure or model predictions (R2MDP) and the concordance correlation coefficient. The accuracy and precision of rumen-degradable N and undegradable N and bacterial N flows were improved with reduced bias. The CNCPS v6.5 predicted accurate and precise milk yield according to the first-limiting nutrient (MP or metabolizable energy) with a R2BLUP=0.97, R2MDP=0.78, and concordance correlation coefficient=0.83. Furthermore, MP-allowable milk was predicted with greater precision than metabolizable energy–allowable milk (R2MDP=0.82 and 0.76, respectively, for MP and metabolizable energy). Results suggest a significant improvement of the model, especially under conditions of MP limitation

Integration of Pharmacogenetics and Pharmacogenomics in Drug Development: Implications for Regulatory and Medical Decision Making in Pediatric Diseases

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Long-term follow-up of patients with type 2 and non-ambulant type 3 spinal muscular atrophy (SMA) treated with olesoxime in the OLEOS trial

In a previous Phase 2 study, olesoxime had a favorable safety profile. Although the primary endpoint was not met, analyses suggested that olesoxime might help in the maintenance of motor function in patients with Types 2/3 SMA. This open-label extension study (OLEOS) further characterizes the safety, tolerability and efficacy of olesoxime over longer therapy durations. In OLEOS, no new safety risks were identified. Compared to matched natural history data, patients treated with olesoxime demonstrated small, non-significant changes in motor function over 52 weeks. Motor function scores were stable for 52 weeks but declined over the remainder of the study. The greatest decline in motor function was seen in patients ≤15 years old, and those with Type 2 SMA had faster motor function decline versus those with Type 3 SMA. Previous treatment with olesoxime in the Phase 2 study was not protective of motor function in OLEOS. Respiratory outcomes were stable in patients with Type 3 SMA >15 years old but declined in patients with Type 2 SMA and in patients with Type 3 SMA ≤15 years old. Overall, with no stabilization of functional measures observed over 130 weeks, OLEOS did not support significant benefit of olesoxime in patients with SMA

Pattern selection as a nonlinear eigenvalue problem

A unique pattern selection in the absolutely unstable regime of driven, nonlinear, open-flow systems is reviewed. It has recently been found in numerical simulations of propagating vortex structures occuring in Taylor-Couette and Rayleigh-Benard systems subject to an externally imposed through-flow. Unlike the stationary patterns in systems without through-flow the spatiotemporal structures of propagating vortices are independent of parameter history, initial conditions, and system length. They do, however, depend on the boundary conditions in addition to the driving rate and the through-flow rate. Our analysis of the Ginzburg-Landau amplitude equation elucidates how the pattern selection can be described by a nonlinear eigenvalue problem with the frequency being the eigenvalue. Approaching the border between absolute and convective instability the eigenvalue problem becomes effectively linear and the selection mechanism approaches that of linear front propagation. PACS: 47.54.+r,47.20.Ky,47.32.-y,47.20.FtComment: 18 pages in Postsript format including 5 figures, to appear in: Lecture Notes in Physics, "Nonlinear Physics of Complex Sytems -- Current Status and Future Trends", Eds. J. Parisi, S. C. Mueller, and W. Zimmermann (Springer, Berlin, 1996

Effect of Cell Age and Membrane Rigidity on Red Blood Cell Shape in Capillary Flow

peer reviewedBlood flow in the microcirculatory system is crucially affected by intrinsic red blood cell (RBC) properties, such as their deformability. In the smallest vessels of this network, RBCs adapt their shapes to the flow conditions. Although it is known that the age of RBCs modifies their physical properties, such as increased cytosol viscosity and altered viscoelastic membrane properties, the evolution of their shape-adapting abilities during senescence remains unclear. In this study, we investigated the effect of RBC properties on the microcapillary in vitro flow behavior and their characteristic shapes in microfluidic channels. For this, we fractioned RBCs from healthy donors according to their age. Moreover, the membranes of fresh RBCs were chemically rigidified using diamide to study the effect of isolated graded-membrane rigidity. Our results show that a fraction of stable, asymmetric, off-centered slipper-like cells at high velocities decreases with increasing age or diamide concentration. However, while old cells form an enhanced number of stable symmetric croissants at the channel centerline, this shape class is suppressed for purely rigidified cells with diamide. Our study provides further knowledge about the distinct effects of age-related changes of intrinsic cell properties on the single-cell flow behavior of RBCs in confined flows due to inter-cellular age-related cell heterogeneity

Pattern selection in the absolutely unstable regime as a nonlinear eigenvalue problem: Taylor vortices in axial flow

A unique pattern selection in the absolutely unstable regime of a driven, nonlinear, open-flow system is analyzed: The spatiotemporal structures of rotationally symmetric vortices that propagate downstream in the annulus of the rotating Taylor-Couette system due to an externally imposed axial through-flow are investigated for two different axial boundary conditions at the in- and outlet. Unlike the stationary patterns in systems without through-flow the spatiotemporal structures of propagating vortices are independent of parameter history, initial conditions, and system's length. They do, however, depend on the axial boundary conditions, the driving rate of the inner cylinder and the through-flow rate. Our analysis of the amplitude equation shows that the pattern selection can be described by a nonlinear eigenvalue problem with the frequency being the eigenvalue. Approaching the border between absolute and convective instability the eigenvalue problem becomes effectively linear and the selection mechanism approaches that one of linear front propagation. PACS:47.54.+r,47.20.Ky,47.32.-y,47.20.FtComment: 15 pages (LateX-file), 8 figures (Postscript

A novel missense variant in ATP11CATP11C is associated with reduced red blood cell phosphatidylserine flippase activity and mild hereditary hemolytic anemia

Adenosine Triphosphatase (ATPase) Phospholipid Transporting 11C gene (ATP11C) encodes the major phosphatidylserine (PS) flippase in human red blood cells (RBCs). Flippases actively transport phospholipids (e.g., PS) from the outer to the inner leaflet to establish and maintain phospholipid asymmetry of the lipid bilayer of cell membranes. This asymmetry is crucial for survival since externalized PS triggers phagocytosis by splenic macrophages. Here we report on pathophysiological consequences of decreased flippase activity, prompted by a patient with hemolytic anemia and hemizygosity for a novel c.2365C > T p.(Leu789Phe) missense variant in ATP11C. ATP11C protein expression was strongly reduced by 58% in patient‐derived RBC ghosts. Furthermore, functional characterization showed only 26% PS flippase activity. These results were confirmed by recombinant mutant ATP11C protein expression in HEK293T cells, which was decreased to 27% compared to wild type, whereas PS‐stimulated ATPase activity was decreased by 57%. Patient RBCs showed a mild increase in PS surface exposure when compared to control RBCs, which further increased in the most dense RBCs after RBC storage stress. The increase in PS was not due to higher global membrane content of PS or other phospholipids. In contrast, membrane lipid lateral distribution showed increased abundance of cholesterol‐enriched domains in RBC low curvature areas. Finally, more dense RBCs and subtle changes in RBC morphology under flow hint toward alterations in flow behavior of ATP11C‐deficient RBCs. Altogether, ATP11C deficiency is the likely cause of hemolytic anemia in our patient, thereby underlining the physiological role and relevance of this flippase in human RBCs

'OH MY GOD! BUY IT!' A multimodal discourse analysis of the discursive strategies used by Chinese ecommerce live-streamer Austin Li

Ecommerce livestreaming, also known as live commerce or social commerce, has taken off over the past two years in East Asia and is showing the tendency of going global. Intrigued by the phenomenal success of ecommerce livestream, we concentrate on analyzing the most prominent and illustrative ex- ample of Chinese ecommerce live-streamer Austin Li. Through this individual case study, we aim to investigate discursive strategies employed in ecommerce livestreaming and reveal resources specific to this new media genre. Guided by multimodal discourse analysis, our research first accommodates the socio-eco- nomic context of Li’s success to warrant social situatedness in interpreting data. After that we move into analyzing his discourse employed in livestreaming. Re- search findings suggest that in attention economy, Li strategically utilizes his male gender as a resource in trying on lipsticks for female customers. His dis- course in multiple modes serves to build consumer trust and propagate products. An in-depth analysis of his discursive strategies indicates that, ecommerce livestreaming as a new form of advertising not only shares commonalities with traditional advertisement discourse but also embodies affordances that are spe- cific to livestreaming platforms. To be more specific, livestreaming is featured with delimitation of time, real-time interactivity, and video-aided communica- tion. These affordances enable Li to adopt more interactive and personalized per- suasive discourse than conventional advertisement