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At least 739 records · Page 41

Two disulfide-reducing pathways are required for the maturation of plastid c -type cytochromes in Chlamydomonas reinhardtii

Abstract In plastids, conversion of light energy into ATP relies on cytochrome f, a key electron carrier with a heme covalently attached to a CXXCH motif. Covalent heme attachment requires reduction of the disulfide-bonded CXXCH by CCS5 and CCS4. CCS5 receives electrons from the oxidoreductase CCDA, while CCS4 is a protein of unknown function. In Chlamydomonas reinhardtii, loss of CCS4 or CCS5 yields a partial cytochrome f assembly defect. Here, we report that the ccs4ccs5 double mutant displays a synthetic photosynthetic defect characterized by a complete loss of holocytochrome f assembly. This defect is chemically corrected by reducing agents, confirming the placement of CCS4 and CCS5 in a reducing pathway. CCS4-like proteins occur in the green lineage, and we show that HCF153, a distant ortholog from Arabidopsis thaliana, can substitute for Chlamydomonas CCS4. Dominant suppressor mutations mapping to the CCS4 gene were identified in photosynthetic revertants of the ccs4ccs5 mutants. The suppressor mutations yield changes in the stroma-facing domain of CCS4 that restore holocytochrome f assembly above the residual levels detected in ccs5. Because the CCDA protein accumulation is decreased specifically in the ccs4 mutant, we hypothesize the suppressor mutations enhance the supply of reducing power through CCDA in the absence of CCS5. We discuss the operation of a CCS5-dependent and a CCS5-independent pathway controlling the redox status of the heme-binding cysteines of apocytochrome f.

Genetics & Heredity↗

Deducing the N- and O-glycosylation profile of the spike protein of novel coronavirus SARS-CoV-2

The current emergence of the novel coronavirus pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) demands the development of new therapeutic strategies to prevent rapid progress of mortalities. The coronavirus spike (S) protein, which facilitates viral attachment, entry and membrane fusion is heavily glycosylated and plays a critical role in the elicitation of the host immune response. The spike protein is comprised of two protein subunits (S1 and S2), which together possess 22 potential N-glycosylation sites. Herein, we report the glycosylation mapping on spike protein subunits S1 and S2 expressed on human cells through high-resolution mass spectrometry. We have characterized the quantitative N-glycosylation profile on spike protein and interestingly, observed unexpected O-glycosylation modifications on the receptor-binding domain of spike protein subunit S1. Even though O-glycosylation has been predicted on the spike protein of SARS-CoV-2, this is the first report of experimental data for both the site of O-glycosylation and identity of the O-glycans attached on the subunit S1. Our data on the N- and O-glycosylation are strengthened by extensive manual interpretation of each glycopeptide spectra in addition to using bioinformatics tools to confirm the complexity of glycosylation in the spike protein. Here, the elucidation of the glycan repertoire on the spike protein provides insights into the viral binding studies and more importantly, propels research toward the development of a suitable vaccine candidate.

59 BASIC BIOLOGICAL SCIENCES↗

Comprehensive characterization of N- and O- glycosylation of SARS-CoV-2 human receptor angiotensin converting enzyme 2

The emergence of the coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has created the need for development of new therapeutic strategies. Understanding the mode of viral attachment, entry and replication has become a key aspect of such interventions. The coronavirus surface features a trimeric spike (S) protein that is essential for viral attachment, entry and membrane fusion. The S protein of SARS-CoV-2 binds to human angiotensin converting enzyme 2 (hACE2) for entry. Herein, we describe glycomic and glycoproteomic analysis of hACE2 expressed in HEK293 cells. We observed high glycan occupancy (73.2 to 100%) at all seven possible N-glycosylation sites and surprisingly detected one novel O-glycosylation site. To deduce the detailed structure of glycan epitopes on hACE2 that may be involved in viral binding, we have characterized the terminal sialic acid linkages, the presence of bisecting GlcNAc and the pattern of N-glycan fucosylation. We have conducted extensive manual interpretation of each glycopeptide and glycan spectrum, in addition to using bioinformatics tools to validate the hACE2 glycosylation. Our elucidation of the site-specific glycosylation and its terminal orientations on the hACE2 receptor, along with the modeling of hACE2 glycosylation sites can aid in understanding the intriguing virus-receptor interactions and assist in the development of novel therapeutics to prevent viral entry. Here, the relevance of studying the role of ACE2 is further increased due to some recent reports about the varying ACE2 dependent complications with regard to age, sex, race and pre-existing conditions of COVID-19 patients.

59 BASIC BIOLOGICAL SCIENCES↗

Mechanisms and models of the turbulent boundary layers at transcritical conditions

Computational models for high-pressure transcritical turbulence in wall-modeled large-eddy simulation typically rely on wall-function models to overcome the need for resolving the boundary layer structures. However, the mechanisms and models of turbulent boundary layer at transcritical conditions remain poorly understood since the near-wall flow and heat transfer are significantly affected by the enhanced fluctuations and steep gradients in thermodynamic properties. Here, to address this issue, we study the mechanisms of transcritical turbulent boundary layers and wall-attached models at transcritical conditions. It is shown that the real-fluid variable-property effects are associated with the coupling between the near-wall cycle and the wall-normal coherent motions in the outer layer, resulting in the amplification of turbulent energy in the outer layer and noticeable energy transfer between the log-layer and the outer layer; hence, turbulence in the log-layer is modulated by the outer layer. Based on this underlying physical principle, we propose the characteristic velocity and length scales for the attached eddy at transcritical conditions, and extend the attached eddy model to transcritical turbulent boundary layers by introducing a mixed scaling that incorporates both inner and outer scalings. We show that the new characteristic scales and extended attached-eddy model perform well in characterizing the structures in transcritical turbulent boundary layers.

Li, Fangbo↗

Umbilical Fatigue Analysis for a Wave Energy Converter

Unique umbilical designs for wave energy converters (WECs), including the ability to handle significantly larger motions and loads over long deployments, are often required when conventional marine umbilical designs for offshore oil and gas and offshore wind may not meet the design and cost needs of wave energy technologies. This study details a fatigue analysis of a dynamic power umbilical attached to a two-body floating point absorber WEC system, using the sea states provided for the PacWave testing facilities. The 6 degrees of freedom motion time history for the WEC was simulated, and the motions of the attachment point for the umbilical on the WEC and respective sea states were used to analyze the dynamic motions and fatigue of the connected power umbilical to predict the fatigue life. The results show that the fatigue damage observed is more significant in shallow water, and extensive fatigue damage may occur because of the larger curvature response of the umbilical. The umbilical configurations departing at 90 deg off incoming waves were found to have the highest fatigue life attributed to less extension or compression of the umbilical. However, additional bend stiffener/limiter features may need to be incorporated into the buoyancy section and touchdown regions to minimize curvature-induced fatigue.

50 EE - Wind and Water Power Program - Water (EE-4↗

Capturing Ultrafast Electron Driven Chemical Reactions in Molecules

The goals of this project are to develop experimental tools to investigate the dynamics of dissociative electron attachment (DEA) reactions in real time. In a DEA reaction, the attachment of a low energy electron to a molecule results in bond breaking and the fragmentation of the molecule. A new instrument has been designed and is under construction to perform time-resolved photoelectron and photo-fragment spectroscopy following electron transfer from an iodide anion to a molecule within a mass-selected molecular dimer. Iodide binds only weakly through the dipole interaction with many polar molecules, and such molecules forming a dimer with iodide undergo only small changes from the equilibrium geometry in isolation. When the electron is transferred from the iodide, the iodine atom is no longer bound, so the electron transfer step can be considered analogous to electron attachment and brings the key advantage of ultrafast timing precision. The experiments proposed here will establish the experimental tools to interrogate the dynamics of dissociative electron attachment on the fundamental timescales of the nuclear motion, by simultaneously probing the valence electronic structure and the fragmentation pathways.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Pose Estimation for the Inspecta Project

In this addendum report we take the lessons learned from our initial generic object pose estimation research described in (Smartt, et al., 2025) and apply it to metal cup seals, which are attached to items after an international nuclear safeguards inspection (i.e., metal cup seals may be attached to containers of spent nuclear fuel after an inspector has measured and verified the material). We have also refined our software pipeline and training techniques since the final Inspecta report and will describe that process here. The objective of this task is to allow a robot to acquire images of the engraved seal characters to confirm that the correct seal is attached to the correct container. However, the seal may be attached in any orientation and therefore the robot manipulator arm with a camera needs to align with the normal face of the seal.

98 NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL P↗

Design of a space shuttle structural dynamics model

A 1/8 scale structural dynamics model of a parallel burn space shuttle has been designed. Basic objectives were to represent the significant low frequency structural dynamic characteristics while keeping the fabrication costs low. The model was derived from the proposed Grumman Design 619 space shuttle. The design includes an orbiter, two solid rocket motors (SRM) and an external tank (ET). The ET consists of a monocoque LO2 tank an interbank skirt with three frames to accept SRM attachment members, an LH2 tank with 10 frames of which 3 provide for orbiter attachment members, and an aft skirt with on frame to provide for aft SRM attachment members. The frames designed for the SRM attachments are fitted with transverse struts to take symmetric loads.

Source record↗

The Space Shuttle orbiter payload retention systems

Payloads are secured in the orbiter payload bay by the payload retention system or are equipped with their own unique retention systems. The orbiter payload retention mechanisms provide structural attachments for each payload by using four or five attachment points to secure the payload within the orbiter payload bay during all phases of the orbiter mission. The payload retention system (PRS) is an electromechanical system that provides standarized payload carrier attachment fittings to accommodate up to five payloads for each orbiter flight. The mechanisms are able to function under either l-g or zero-g conditions. Payload berthing or deberthing on orbit is accomplished by utilizing the remote manipulator system (RMS). The retention mechanisms provide the capability for either vertical or horizontal payload installation or removal. The payload support points are selected to minimize point torsional, bending, and radial loads imparted to the payloads. In addition to the remotely controlled latching system, the passive system used for nondeployable payloads performs the same function as the RMS except it provides fixed attachments to the orbiter.

Hardee, J. H.↗

Energy Efficient High-Pressure Turbine Leakage Technology Report

The leakage test program was one of such supporting technology programs structured to provide guidance to the Energy Efficient Engine High Pressure Turbine Component Design Effort. Leakage reduction techniques were identified and evaluated. Test models were used to simulate component leak paths and to evaluate leakage reduction techniques. These models simulated the blade/disk attachment, the vane inner platform attachment, and the vane outer platform attachment combined with the blade outer airseal. Disk blade attachment testing indicated that leakage in this area could be reduced to very low levels by paying careful attention to the tolerances along the contact surface between the blade vibration damper and the blade platform contact surface. The aim of feather seal testing was to achieve a goal for an effective leakage gap of one mil (.001 inch) per inch of feather seal length. Results indicated that effective gaps even below the goal level were achievable by (1) maintaining close tolerances between feather seals and their slots to minimize end gaps and limit seal rotation, (2) avoiding feather seal overlap, and (3) minimizing feather seal intersections. W seals were shown to be effective leakage control devices. Wire rope, in its present state of development, was shown not to be an effective sealing concept for application to the component design.

Gardner, W. B.↗

Ladder supported ring bar circuit

An improved slow wave circuit especially useful in backward wave oscillators includes a slow wave circuit in a waveguide. The slow wave circuit is comprised of rings disposed between and attached to respective stubs. The stubs are attached to opposing sidewalls of the waveguide. To the end that opposed, interacting magnetic fields will be established to provide a very high coupling impedance for the slow wave structure, axially orientated bars are connected between rings in alternate spaces and adjacent to the attachment points of stubs. Similarly, axial bars are connected between rings in the spaces which do not include bars and at points adjacent to the attachment of bars.

Kosmahl, H. G.↗

Nonlinear aerodynamic effects on bodies in supersonic flow

The supersonic flow about generic bodies was analyzed to identify the elments of the nonlinear flow and to determine the influence of geometry and flow conditions on the magnitude of these nonlinearities. The nonlinear effects were attributed to separated-flow nonlinearities and attached-flow nonlinearities. The nonlinear attached-flow contribution was further broken down into large-disturbance effects and entropy effects. Conical, attached-flow bundaries were developed to illustrate the flow regimes where the nonlinear effects are significant, and the use of these boundaries for angle of attack and three-dimensional geometries was indicated. Normal-force and pressure comparisons showed that the large-disturbance and separated-flow effects were the dominant nonlinear effects at low supersonic Mach numbers and that the entropy effects were dominant for high supersonic Mach number flow. The magnitude of all the nonlinear effects increased with increasing angle of attack. A full-potential method, NCOREL, which includes an approximate entropy correction, was shown to provide accurate attached-flow pressure estimates from Mach 1.6 through 4.6.

Pittman, J. L.↗

Selectable towline spin chute system

An emergency spin recovery parachute is presented that is housed within a centrally mounted housing on the aft end of an aircraft and connected to a ring fitting within the housing. Two selectively latching shackles connected to separate towlines are openly disposed adjacent the ring fitting. The towlines extend in opposite directions from the housing along the aircraft wing to attachment points adjacent the wing-tips where the other end of each towline is secured. Upon pilot command, one of the open shackles latches to the ring fitting to attach the towline connected thereto, and a second command signal deploys the parachute. Suitable break-away straps secure the towlines to the aircraft surface until the parachute is deployed and the resulting force on the towline attached to the parachute overcomes the straps and permits the towline to extend to the point of attachment to exert sufficient drag on the spinning aircraft to permit the pilot to regain control of the aircraft. To employ the parachute as a drag chute to reduce landing speeds, both shackles and their respective towlines are latched to the ring fitting.

Vairo, Daniel M.↗

End effector with astronaut foot restraint

The combination of a foot restraint platform designed primarily for use by an astronaut being rigidly and permanently attached to an end effector which is suitable for attachment to the manipulator arm of a remote manipulating system is described. The foot restraint platform is attached by a brace to the end effector at a location away from the grappling interface of the end effector. The platform comprises a support plate provided with a pair of stirrups for receiving the toe portion of an astronaut's boots when standing on the platform and a pair of heel retainers in the form of raised members which are fixed to the surface of the platform and located to provide abutment surfaces for abutting engagement with the heels of the astronaut's boots when his toes are in the stirrups. The heel retainers preclude a backward sliding movement of the feet on the platform and instead require a lifting of the heels in order to extract the feet. The brace for attaching the foot restraint platform to the end effector may include a pivot or swivel joint to permit various orientations of the platform with respect to the end effector.

Monford, Leo G., Jr.↗

Quick-connect fasteners for assembling devices in space

A quick-connect fastener of a relatively-simple straightforward design is arranged with a tubular body adapted to be engaged against an attachment fitting in coincidental alignment with an opening in that fitting. A tubular collet having flexible finger projecting from its forward end is arranged in the fastener body to be shifted forwardly by an elongated expander member coaxially arranged within the tubular collet for advancing the collet fingers into the opening in the attachment fitting. Biasing means are arranged between the elongated expander member and a rotatable actuator which is threadedly mounted within the tubular collet so as to be rotated for urging the expander member into engagement with the collet fingers. A first coupling member is arranged on the rotatable actuator to be accessible from outside of the fastener so that a second coupling member on the distal end of a flexible shaft can be introduced into the fastener body and coupled to the first coupling member to enable a typical actuating tool coupled to the shaft outside of the fastener body to be operated for advancing the outwardly-enlarged ends of the collet fingers into the opening in the attachment fitting and thereafter rotating the actuator member to expand the fingers to that attachment fitting. Upon expansion of the collet fingers, the biasing means impose a biasing force on the expander to releasably retain the fingers in their latching positions.

Wesselski, Clarence J.↗

Assured crew return vehicle post landing configuration design and test

The 1991-1992 senior Mechanical and Aerospace Engineering Design class continued work on the post landing configurations for the Assured Crew Return Vehicle (ACRV) and the Emergency Egress Couch (EEC). The ACRV will be permanently docked to Space Station Freedom fulfilling NASA's commitment of Assured Crew Return Capability in the event of an accident or illness aboard Space Station Freedom. The EEC provides medical support and a transportation surface for an incapacitated crew member. The objective of the projects was to give the ACRV Project Office data to feed into their feasibility studies. Four design teams were given the task of developing models with dynamically and geometrically scaled characteristics. Groups one and two combined efforts to design a one-fifth scale model for the Apollo Command Module derivative, an on-board flotation system, and a lift attachment point system. This model was designed to test the feasibility of a rigid flotation and stabilization system and to determine the dynamics associated with lifting the vehicle during retrieval. However, due to priorities, it was not built. Group three designed a one-fifth scale model of the Johnson Space Center (JSC) benchmark configuration, the Station Crew Return Alternative Module (SCRAM) with a lift attachment point system. This model helped to determine the flotation and lifting characteristics of the SCRAM configuration. Group four designed a full scale EEC with changeable geometric and geometric and dynamic characteristics. This model provided data on the geometric characteristics of the EEC and on the placement of the CG and moment of inertia. It also gave the helicopter rescue personnel direct input to the feasibility study. Section 1 describes in detail the design of a one-fifth scale model of the Apollo Command Module Derivative (ACMD) ACRV. The objective of the ACMD Configuration Model Team was to use geometric and dynamic constraints to design a one-fifth scale working model of the Apollo Command Module Derivative (ACMD) configuration with a Lift Attachment Point (LAP) System. This model was required to incorporate a rigidly mounted flotation system and the egress system designed the previous academic year. The LAP system was to be used to determine the dynamic effects of locating the lifting points at different locations on the vehicle. The team was then to build and test the model; however, due to priorities, this did not occur. To better simulate the ACMD after a water landing, the nose cone section was removed and the deck area exposed. The areas researched during the design process were construction, center of gravity and moment of inertia, and lift attachment points.

Source record↗

Developing a control system for ARES 2

A great deal of analysis and testing is conducted at the NASA Langley Research Center to support the development of safe and reliable helicopter rotor systems. This work is performed by the Rotorcraft Aeroelasticity Group located in the Transonic Dynamics Tunnel (TDT) facility. Over the past two decades a wide variety of tests have been successfully conducted in the TDT and their results have contributed significantly to the understanding of aeromechanical phenomena in rotor systems. This has led to improved tools for analysis and design, and ultimately to the development, of improved rotor systems. The TDT facility is ideally suited for these tests due to its unique ability to use a heavy gas as a working medium. This allows the model to be scaled such that the results obtained may be readily extrapolated to full scale. Until recently, the rotor system to be tested has been mounted on a fixed balance which is attached to the longeron which is attached to the stand through a single pitching degree of freedom. The testbed used is known as the Aeroelastic Rotor Experimental System (ARES 1). In order to extend the experimental capabilities to investigate the full rotor/body dynamic coupling present in a rotorcraft, a very ambitious project has been undertaken to design and construct a six degree of freedom system that can be controlled so as to emulate the inertial characteristics of a prescribed model fuselage. The electronic and mechanical hardware for this system has already been designed and constructed. This system is known ar ARES II. The rotor and its drive system are mounted on the balance which is attached to the longeron via six hydraulic actuators. This six degree of freedom parallel linkage is referred to in the literature as a Stuart Platform. By properly adjusting the length of the hydraulic actuators it is possible to position and orient the balance relative to the longeron. The longeron is attached to the stand via a pitch degree of freedom to allow testing over various forward flight regimes. One major task remaining to complete this testbed is the design and synthesis of a control system. To do this properly requires an understanding of the kinematics and dynamics of the system and robust control design. A brief description of the development of a control design is given.

Fitzsimons, Philip M.↗

Active plate and wing research using EDAP elements

The deflection characteristics of structures using directionally attached piezoelectric (DAP) and enhanced DAP (EDAP) elements are investigated analytically and experimentally. It is shown that directional attachment can be achieved by partial attachment, transverse shear lag, and differential stiffness bonding. The effective stiffnesses of DAP elements can be accurately modeled by an active element area estimation of partial attachment with a shear lag modification. Methods of integrating DAP and EDAP elements in missile wings and ways of alleviating such difficulties as control power degradation and adverse coupling are discussed.

Barrett, Ron↗