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At least 451 records · Page 25

Automated instant labeling chemistry workflow for real-time monitoring of monoclonal antibody N -glycosylation

With the transition toward continuous bioprocessing, process analytical technology (PAT) is becoming necessary for rapid and reliable in-process monitoring during biotherapeutics manufacturing. Bioprocess 4.0 is looking to build end-to-end bioprocesses that include PAT-enabled real-time process control. This is especially important for drug product quality attributes that can change during bioprocessing, such as protein N-glycosylation, a critical quality attribute for most monoclonal antibody (mAb) therapeutics. Glycosylation of mAbs is known to influence their efficacy as therapeutics and is regulated for a majority of mAb products on the market today. Currently, there is no method to truly measure N-glycosylation using on-line PAT, hence making it impractical to design upstream process control strategies. We recently described the N-GLYcanyzer workflow: an integrated PAT unit that measures mAb N-glycosylation within 3 hours of automated sampling from a bioreactor. Here, we integrated Agilent's Instant Procainamide (InstantPC) based chemistry workflow into the N-GLYcanyzer PAT unit to allow for nearly 10× faster near real-time analysis of mAb glycoforms. Furthermore, our methodology is explained in detail to allow for replication of the PAT workflow as well as present a case study demonstrating the use of this PAT to autonomously monitor a mammalian cell perfusion process at the bench scale to gain increased knowledge of mAb glycosylation dynamics during continuous biologics manufacturing using Chinese hamster ovary (CHO) cells.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Towards the Flexibility of HVDC-Interconnected Systems: A Novel Emergency Freqeuncy Response Model

In this paper, we propose a novel multi-time scale emergency frequency response model by unlocking the flexibility of High Voltage Direct Current (HVDC) systems. Unlike assigning power ramping rates for FACTS to regulate frequency in traditional methods, this paper designs a step-change electromagnetic power frequency response (EPFR) scheme, by leveraging the temporal over/under DC voltage capability of HVDC. Wherein the Kullback-Leibler Divergence is adopted to convexify the modified swing equation after integrating the step-change power. Further, to avoid the complicated differential equations, we equivalently reformulate the duration limits of DC voltage deviation into the HVDC decreasing power ramping rates, which participate in the system primary frequency response. Finally, the new steady-state operation level of HVDC is involved with the secondary frequency response. As a result, an improved three-level algorithm is developed to solve the model, wherein the instant step-change, primary, and secondary frequency response are coordinated together. After applying the proposed frequency response scheme on the test system, the EPFR is validated to effectively provide the most instantaneous supports when faced with bulk power loss due to extreme contingencies, and the resilience is ensured within acceptable expenditures.

Jiang, Sufan↗

Enabling Grid-Forming Control with Fault Ride-Through in Unbalanced Distribution Networks

Distribution networks are often unbalanced, causing oscillatory responses in inverter control designed for balanced conditions. Here, to address this problem, this paper proposes a novel time-domain transformation appropriate for inverter control and enables the decomposition of three-phase unbalanced signals into constant positive and negative components. Relations useful for calculating unbalanced active and reactive power are derived from first principle, providing insight into vector products of unbalanced three-phase signals. Furthermore, a grid-forming control effective under unbalanced conditions is developed, which delivers superior performance while meeting UNIFI1 specifications for grid-forming control under unbalanced conditions. specifications applicable to category 4 inverter-based resource, like setting and regulating frequency/voltage, providing voltage support, sharing active power, injecting negative sequence current, and riding through faults. A current limiter is proposed for safe fault ride-through and integrates with the grid-forming control featuring frequency/voltage droop controllers and current and voltage control loops. The transformation of interconnected inverters is formulated and stability of the proposed control analyzed to support robust parameter selections. The effectiveness of the proposed transformation and grid-forming control is demonstrated through analytical results and real-time simulation of a IEEE 123 distribution network on the Real-Time Digital Simulator. Comparison with existing methods shows that the proposed strategy satisfies the UNIFI specifications with a much better performance.

24 - POWER TRANSMISSION AND DISTRIBUTION↗

Integrating Transactive Energy into Reliability Evaluation for a Self-healing Distribution System with Microgrid

Non-utility owned distributed energy resources (DERs) are mostly untapped currently, but they can provide many grid services such as voltage regulation and service restoration, if properly controlled, and can improve the distribution systems reliability when coordinated with utility-owned assets such as self-healing control and microgrids. This paper integrates transactive energy control into the distribution system reliability evaluation to quantitatively assess the impact of non-utility owned DERs on reliability improvement. Here, a transactive reactive power control strategy is designed to incentivize the DERs to provide reactive power support for improving voltage profiles thus enabling additional customer load restoration during an outage. Also, an operational sequence to coordinate the non-utility owned DERs with the utility owned self-healing control and utility owned microgrids is designed and integrated into the service restoration process with the operational constraints guaranteed by checking the three-phase unbalanced power flow for post-fault network reconfiguration. The reliability indices are then calculated through a Monte Carlo simulation. The transactive reactive power control strategy is tested on a four-feeder distribution system operated by Duke Energy in the U.S. Results demonstrate that the non-utility owned DERs with the transactive control improve the reliability of both the system and critical loads by more than 30%.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Involute Working Group – FSI Analysis of Fuel Plates Using Finite Volume and Finite Element Methods

The three involute plate research reactors RHF, HFIR, and FRM II have expressed an interest in using computational software to carry their steady-state safety analysis. Since these tools represent a significant departure from the methods used currently (one-dimensional), the acceptability of the new approach by regulators requires thorough verification and validation of these tools. Therefore, Argonne National Laboratory and the three involute-plate reactors formed an informal group called the Involute Working Group aiming at qualifying computational tools to perform steady-state safety analysis. The present report focuses on a comparison of finite volume and finite element methods to model solids in fluid-structure interaction problems with the goal to estimate the coolant flow-induced fuel plate deflections obtained with the two methods. The finite volume method will be obsoleted in STARCCM+ by the end of 2021, nevertheless, this evaluation is important because the method was used by ANL researchers to model the response of the fuel plates, despite its drawbacks, which are discussed in the report. It was essential to check how those estimates compare to the results obtained with the finite element method that is considered superior for structural analysis. Various geometries, i.e., flat, cylindrical and circle-involute fuel plates, as well as coolant flow speed, were considered. The comparison shows that, independently of the plate geometry, the finite volume method significantly underestimates the deflection as compared to finite element method for coarser meshes. When the discretization is developed as a result of a mesh sensitivity study using finite element method, the result obtained using finite volume method can be a few times smaller than the corresponding finite element method solution. A code-to-code comparison , between STAR-CCM+ and LS-DYNA was included in the analysis. Within the LS-DYNA models, two types of finite element formulations were used: solid and shell finite elements. Mesh sensitivity study showed that both approaches converge to a similar value that was obtained with STAR-CCM+ finite element solver. The evaluation of the computational solvers was extended by adding two benchmark cases from the STAR-CCM+ Verification Suite and presented in the Appendix A. The selected cases are: (1) bending of a cantilever beam under external load, and (2) cylindrical shell deformation analysis, known in the literature as ‘Scordelis-Lo roof’. The problems were solved with finite volume, and finite element methods, and the results confirmed the previously discussed findings. The analysis shows that the finite element solver is superior to the finite volume solver in terms of representation of model geometry and estimating the structural behavior of fuel plates. Depending on the ratio of the load to the flexibility of the plate, the finite volume solver can greatly under- or overestimate the structural response if a very carefully selected mesh is not used.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

Line-oriented flight training: Northwest Airlines

An exemption from certain FAA regulations which stereotype simulator flight training was obtained and pilots with current line experience were used to prepare and develop scenarios for a program in which each crew member would be trained to recognize and properly use all available resouces. The development of the scenarios for training to proficiency and pilot reaction to the training sessions are discussed.

Nunn, H. T.↗

INTESPACE's new thermal-vacuum test facility: SIMMER

The development of an European satellite market over the last 10 years, the industrialization of space applications, and the new requirements from satellite prime contractors have led INTESPACE to increase the test center's environmental testing capacities through the addition of a new thermal-vacuum test facility of impressive dimensions referred to as the SIMMER. The SIMMER is a simulator specifically created for the purpose of conducting acceptance tests of satellites and of large structures of the double launching ARIANE IV or half ARIANE V classes. The chamber is 8.3 meters long with a diameter of 10 meters. The conceptual design of a chamber in the horizontal plane and at floor level is in a view to simplify test preparation and to permit final electrical checks of the spacecraft in its actual test configuration prior to the closing of the chamber. The characteristics of the SIMMER complies with the requirements being currently defined in terms of thermal-vacuum tests: (1) thermal regulation (temperatures cycling between 100 K and 360 K); (2) clean vacuum (10(exp -6) mbar); (3) 600 measurement channels; and (4) 100 000 cleanliness class. The SIMMER is located in INTESPACE's space vehicle test complex in which a large variety of environmental test facilities are made available for having a whole test program completed under one and a same roof.

Duprat, Raymond↗

Experimental Modification of Rat Pituitary Prolactin Cell Function During and After Spaceflight

Experimental modification of rat pituitary prolactin cell function during and after spaceflight. This study was done to evaluate the effects of microgravity on prolactin (PRL) cells of the male rat pituitary gland. We used the identical passive closed-vial cell culture system that was described for the culture of growth hormone cells (W C. Hymer, R. E. Grindeland, T. Salada, P. Nye, E. Grossman, and R Lane). After an 8-day spaceflight, all flight media (containing released PRL), as well as extracts (containing intracellular PRL), contained significantly lower amounts of immunoreactive PRL than their corresponding ground control samples. On the other hand, these same samples, when assessed for their biological activities by two different in vitro lymphocyte assays, yielded disparate results that may reflect posttranslational modifications to the hormone molecule. Other data showed that: (1) the apparent molecular weights of released PRL molecules were not altered by microgravity; but (2) the region from which the PRL cells came (dorsal or ventral) made a significant difference in the amount and activity of PRL released from the flight cells. Because there is much current interest in the role that PRL may play in the regulation of the immune system and because changes in both cellular and humoral immunity accompany spaceflight, this study could help define future microgravity research in this area.

Hymer, W. C.↗

Explosive Fracturing of an F-16 Canopy for Through-Canopy Crew Egress

Through-canopy crew egress, such as in the Harrier (AV-8B) aircraft, expands escape envelopes by reducing seat ejection delays in waiting for canopy jettison. Adverse aircraft attitude and reduced forward flight speed can further increase the times for canopy jettison. However, the advent of heavy, high-strength polycarbonate canopies for bird-strike resistance has not only increased jettison times, but has made seat penetration impossible. The goal of the effort described in this paper was to demonstrate a method of explosively fracturing the F-16 polycarbonate canopy to allow through-canopy crew ejection. The objectives of this effort were to: 1. Mount the explosive materials on the exterior of the canopy within the mold line, 2. Minimize visual obstructions, 3. Minimize internal debris on explosive activation, 4. Operate within less than 10 ms, 5. Maintain the shape of the canopy after functioning to prevent major pieces from entering the cockpit, and 6. Minimize the resistance of the canopy to seat penetration. All goals and objectives were met in a full-scale test demonstration. In addition to expanding crew escape envelopes, this canopy fracture approach offers the potential for reducing system complexity, weight and cost, while increasing overall reliability, compared to current canopy jettison approaches. To comply with International Traffic in Arms Regulations (ITAR) and permit public disclosure, this document addresses only the principles of explosive fracturing of the F-16 canopy materials and the end result. ITAR regulations restrict information on improving the performance of weapon systems. Therefore, details on the explosive loads and final assembly of this canopy fracture approach, necessary to assure functional performance, are not included.

Bement, Laurence J.↗

Physiological Maturation of Regenerating Hair Cells

The bullfrog saccule, a sensor of gravity and substrate-borne vibration, is a model system for hair cell transduction. Saccular hair cells also increase in number throughout adult life and rapidly recover after hair cell damage, making this organ an ideal system for studying hair cell development, repair, and regeneration. We have used of hair cell and supporting cell immunocytochemical markers to identify damaged hair cells and hair cell precursors in organotypic cultures of the bullfrog saccule. We then used an innovative combination of confocal, electron, and time-lapse microscopy to study the fate of damaged hair cells and the origin of new hair cells after gentamicin ototoxicity in normal and mitotically blocked saccular cultures. These studies have shown that gentamicin ototoxicity produces both lethal and sublethal hair cell damage. They have also shown that hair cell recovery in this organ takes place by both the repair of sublethally damaged hair cells and by the replacement of lost hair cells by mitotic regeneration. In parallel studies, we have used biophysical and molecular biological techniques to study the differentiation and innervation of developing, repairing, and regenerating hair cells. More specifically, we have used RT-PCR to obtain the bullfrog homologues of L-type voltage- gated calcium (L-VGCC) and large-conductance Ca(2+)-activated potassium (BK) channel genes. We have then obtained probes for these genes and, using in situ hybridization, begun to examine their expression in the bullfrog saccule and amphibian papilla. We have also used fluorescent-labeled channel toxins and channel toxin derivatives to determine the time of appearance of L-type voltage-gated calcium (L-VGCC) and Ca(2+)-activated potassium (BK) channels and to study dynamic changes in the number, distribution, and co-localization of these proteins in developing, repairing, and regenerating hair cells. Using time-lapse microscopy, we are also studying the dynamic relationship between ion channel clustering and synaptic formation in hair cells and afferent neurons. In future studies, we will determine when hair cell precursors acquire electrical tuning, and, using whole-cell patch-clamp techniques, identify and characterize their L-VGCC and BK currents. We will also use biophysical techniques to determine the number of L-VGCC and BK channels and the size and gating kinetics of their underlying L-VGCC and BK conductances, correlating these variables with the amplitude and frequency of membrane oscillations produced by intracellular current steps. We expect these studies to determine how hair cells regulate ion channel expression to achieve specific physiological responses.

Baird, Richard A.↗

Inlet Radiated Noise Predictions for the NASA Source Diagnostic Test Fan Using Physics-Based Simulations

Commercial supersonic aircraft are expected to produce considerably more noise during landing and take-off operations than existing subsonic aircraft. Establishing noise regulation is challenging because no examples of this aircraft type are currently flying. Existing aircraft noise prediction tools can be used, but the noise source models for these empirical tools are typically based on data from subsonic aircraft components. The present work documents a procedure for using physics-based computational tools to generate the propagating sound field radiating out the nacelle inlet. Although no analytical source modeling is demonstrated, these tools utilize measured data to compare the complex pressure for various propagating modes. The discrepancies found in applying the propagation tools to the subsonic inlet will be ascribed to calculations of supersonic inlets being used to update current empirical noise prediction tools.

Noise Predictions↗

Dose-Response Data Collection Preparation for the NASA Quesst Mission With the X-59

Supersonic flight has the potential to cut travel times in half; however, community annoyance to sonic booms led to the ban on overland commercial supersonic flight by the Federal Aviation Administration in 1973. As part of its Quesst mission, NASA is developing the X-59 aircraft to demonstrate quiet supersonic flight and collect dose-response data to inform regulators in their efforts to establish noise standards to replace the current supersonic speed limit. This talk presents an overview of the NASA Quesst mission and poses some challenges in collecting and analyzing X-59 dose-response data. For the upcoming community flight campaign, the noise dose is the loudness of a supersonic overflight of the X-59 and the perceptual response from participants is captured via surveys after the flyover events. Establishing a relationship between noise dose and perceptual response will be a key product of these efforts.

X-59↗

A rich and bountiful harvest: Key discoveries in plant cell biology

Abstract The field of plant cell biology has a rich history of discovery, going back to Robert Hooke’s discovery of cells themselves. The development of microscopes and preparation techniques has allowed for the visualization of subcellular structures, and the use of protein biochemistry, genetics, and molecular biology has enabled the identification of proteins and mechanisms that regulate key cellular processes. In this review, seven senior plant cell biologists reflect on the development of this research field in the past decades, including the foundational contributions that their teams have made to our rich, current insights into cell biology. Topics covered include signaling and cell morphogenesis, membrane trafficking, cytokinesis, cytoskeletal regulation, and cell wall biology. In addition, these scientists illustrate the pathways to discovery in this exciting research field.

59 BASIC BIOLOGICAL SCIENCES↗

The Landscape of RNA-Protein Interactions in Plants: Approaches and Current Status

RNAs transmit information from DNA to encode proteins that perform all cellular processes and regulate gene expression in multiple ways. From the time of synthesis to degradation, RNA molecules are associated with proteins called RNA-binding proteins (RBPs). The RBPs play diverse roles in many aspects of gene expression including pre-mRNA processing and post-transcriptional and translational regulation. In the last decade, the application of modern techniques to identify RNA–protein interactions with individual proteins, RNAs, and the whole transcriptome has led to the discovery of a hidden landscape of these interactions in plants. Global approaches such as RNA interactome capture (RIC) to identify proteins that bind protein-coding transcripts have led to the identification of close to 2000 putative RBPs in plants. Interestingly, many of these were found to be metabolic enzymes with no known canonical RNA-binding domains. Here, we review the methods used to analyze RNA–protein interactions in plants thus far and highlight the understanding of plant RNA–protein interactions these techniques have provided us. We also review some recent protein-centric, RNA-centric, and global approaches developed with non-plant systems and discuss their potential application to plants. We also provide an overview of results from classical studies of RNA–protein interaction in plants and discuss the significance of the increasingly evident ubiquity of RNA–protein interactions for the study of gene regulation and RNA biology in plants.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Can the ionosphere regulate magnetospheric convection.

A simple model is outlined that relates the dayside magnetopause displacement to the currents feeding the polar cap ionosphere, from which the ionospheric electric field and the flux return rate may be estimated as a function of magnetopause displacement. Then, flux conservation arguments make possible an estimate of the time scale on which convection increases.

Coroniti, F. V.↗

Crowding-Induced Spatial Organization of Gene Expression in Cell-Sized Vesicles

Cell-free protein synthesis is an important tool for studying gene expression and harnessing it for applications. In cells, gene expression is regulated in part by the spatial organization of transcription and translation. Unfortunately, current cell-free approaches are unable to control the organization of molecular components needed for gene expression, which limits the ability to probe and utilize its effects. In this work, we show, using complementary computational and experimental approaches, that macromolecular crowding can be used to control the spatial organization and translational efficiency of gene expression in cell-sized vesicles. Computer simulations and imaging experiments reveal that, as crowding is increased, DNA plasmids become localized at the inner surface of vesicles. Ribosomes, in contrast, remain uniformly distributed, demonstrating that crowding can be used to differentially organize components of gene expression. We further carried out cell-free protein synthesis reactions in cell-sized vesicles and quantified mRNA and protein abundance. At sufficiently high levels of crowding, we observed localization of mRNA near vesicle surfaces, a decrease in translational efficiency and protein abundance, and anomalous scaling of protein abundance as a function of vesicle size. These results are consistent with high levels of crowding causing altered spatial organization and slower diffusion. Our work demonstrates a straightforward way to control the organization of gene expression in cell-sized vesicles and provides insight into the spatial regulation of gene expression in cells.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Economic evaluation of variable renewable energy participation in U.S. ancillary services markets

Variable renewable energy (VRE) is not yet meaningfully participating in U.S. ancillary services (AS) markets. VRE participation in AS markets could provide a new source of revenue for VRE resource owners to offset declining energy and capacity values and a new tool for power system operators to address emerging system constraints. This paper uses a price-taker dispatch model and historical prices to estimate the economic value of standalone and hybrid (battery-paired) VRE participation in AS markets, from the resource owner and electricity system perspectives, in each of the seven U.S. independent system operator and regional transmission organization (ISO/RTO) markets where ancillary service prices are set. Across ISO/RTO markets, average (2015–2019) simulated incremental revenues from power regulation market participation were 0.0–2.9 USD/MWh (+0–15% of revenue without participation) for standalone VRE owners and 1–33 USD/MWh (+1–69%) for hybrid VRE owners. However, ISO/RTO reserve markets are relatively thin and have the potential to become saturated by energy storage projects that are currently in ISO/RTO interconnection queues. In most markets, standalone and hybrid VRE could provide power regulation reserves during periods with high power regulation prices, suggesting that VRE participation in AS markets could have high system value. The analysis highlights the relevance of separate upward and downward power regulation products and indicates that ISOs/RTOs might consider initially focusing on enabling hybrid VRE provision of AS.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗