Search NASA⌕ Search

SEARCH · Search NASA

Results for “study protocol”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 55 records · Page 3

Calibration and data collection protocols for reliable lattice parameter values in electron pair distribution function (ePDF) studies

We explore and describe different protocols for calibrating electron pair distribution function (ePDF) measurements for quantitative studies on nano-materials. We find the most accurate approach to determine the camera-length is to use a standard calibration sample of Au nanoparticles from National Institute of Standards and Technology. Different protocols for data collection are also explored, as are possible operational errors, to find the best approaches for accurate data collection for quantitative ePDF studies.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Proof-of-concept studies of novel protocols for producing highly pure 48 V from a 48 Cr/ 48 V generator

Here, the quest to improve the quality of nuclear data, such as half-lives, transition yields, and reaction cross-sections, is a shared endeavor among various areas of nuclear science. 48 V is a vanadium isotope for which experimental data on neutron reaction cross-sections is needed. However, traditional isotope production techniques cannot produce 48 V with high enough isotopic purity for some of these measurements. “Isotope harvesting” at the Facility for Rare Isotope Beams (FRIB) is a new isotope production technique that could potentially yield 48 V with the necessary purity for such studies. In this case, 48Cr would be collected and allowed to generate 48 V that can be separated from undecayed 48 Cr to yield highly pure 48 V. Thus, any protocol for producing pure 48 V via isotope harvesting would involve utilizing a separation technique that can effectively separate 48 Cr and 48 V. In this study, the radiotracers 51 Cr and 48 V were used to develop possible radiochemical separation methodologies, which can be translated to obtain high purity 48 V via this novel isotope production method. The developed protocols utilize either ion exchange or extraction chromatographic resins. Separations of 51 Cr and 48 V with AG 1-X8 anion exchange resin respectively resulted in recoveries of 95.6(26)% and 96.2(12)% with radionuclidic purities of 92(2)% and 99(1)%. An even more effective Cr and V separation was obtained with an extraction chromatographic resin (TRU resin) and 10 M HNO 3 loading solution. Here, 51 Cr and 48 V respectively had recoveries of 94.1(28)% and 96.2(13)% with high radionuclidic purities (100(2)% and 100(1)%) in small volumes (8.81(8) mL and 5.39(16) mL). This study suggests that, to maximize the yield and isotopic purity of 48 V, the best production protocol would involve utilizing two separations with TRU resin and 10 M HNO 3 to isolate 48 Cr and purify the generated 48 V.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗

SU-E-I-57: Evaluation and Optimization of Effective-Dose Using Different Beam-Hardening Filters in Clinical Pediatric Shunt CT Protocol

Purpose: Study image optimization and radiation dose reduction in pediatric shunt CT scanning protocol through the use of different beam-hardening filters Methods: A 64-slice CT scanner at OU Childrens Hospital has been used to evaluate CT image contrast-to-noise ratio (CNR) and measure effective-doses based on the concept of CT dose index (CTDIvol) using the pediatric head shunt scanning protocol. The routine axial pediatric head shunt scanning protocol that has been optimized for the intrinsic x-ray tube filter has been used to evaluate CNR by acquiring images using the ACR approved CT-phantom and radiation dose CTphantom, which was used to measure CTDIvol. These results were set as reference points to study and evaluate the effects of adding different filtering materials (i.e. Tungsten, Tantalum, Titanium, Nickel and Copper filters) to the existing filter on image quality and radiation dose. To ensure optimal image quality, the scanner routine air calibration was run for each added filter. The image CNR was evaluated for different kVps and wide range of mAs values using above mentioned beam-hardening filters. These scanning protocols were run under axial as well as under helical techniques. The CTDIvol and the effective-dose were measured and calculated for all scanning protocols and added filtration, including the intrinsic x-ray tube filter. Results: Beam-hardening filter shapes energy spectrum, which reduces the dose by 27%. No noticeable changes in image low contrast detectability Conclusion: Effective-dose is very much dependent on the CTDIVol, which is further very much dependent on beam-hardening filters. Substantial reduction in effective-dose is realized using beam-hardening filters as compare to the intrinsic filter. This phantom study showed that significant radiation dose reduction could be achieved in CT pediatric shunt scanning protocols without compromising in diagnostic value of image quality.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

CRCNS US-France Research Proposal: Collaborative Research: Encoding reward expectation in Drosophilia

The fruit fly Drosophila melanogaster has been a valuable model for investigating the genetic and neural bases that underlie learning and memory. Early and most current studies use basic behavior conditioning protocols to study learning in controlled laboratory settings. More recently, the ability to transgenically manipulate many of the brain neurons in the fruit fly with exquisite specificity, and the recent knowledge of the synaptic ‘connectome’ of the fruit fly brain, makes these animals almost unique as a comprehensive model for studies of learning, memory and motivated behavior. In fact, the connectome has revealed many types of new connections that had until now been overlooked. Within this context, the thesis of this proposal is that studies of learning and memory will be greatly enhanced by using more sophisticated means for evaluating memory representations, such as have been developed in vertebrates, and combining those studies with information from the connectome guided by computational modelling. We propose to push beyond the boundaries of existing conditioning protocols for fruit flies to investigate more complex memory representations. In particular, we will investigate the function of reinforcement pathways in relation to the absence of expected reinforcement. More specifically, we propose a series of experiments designed to investigate the memory representations in fruit flies when an expected consequence of a Conditioned Stimulus (CS) fails to occur. Although studies have evaluated how this failure can establish extinction memory for the CS, our studies will go beyond studying extinction. Specifically, we predict that in Drosophila when a CS is associated with a failed expectation of an appetitive food reinforcement it will acquire aversive value, and vice versa for a failed expectation of an aversive reinforcer. We combine these studies with manipulations of reinforcement pathways in the CNS inspired from the connectome, iteratively knitted in with established computational models. Intellectual Merit: The concept of reinforcement expectation and incentive contrast have been influential in the development of studies of associative learning in mammals. These questions are particularly challenging to answer in vertebrates because they require exquisite cellular, temporal, and genetic specificity of experimental manipulations. The recent development of work with identified neurons and their connectomes makes the larval and adult fly brains ripe as models for pushing our understanding of neural bases for these higher- order conditioning phenomena. Broader Impacts: Public health: These analyses and the conceptual framework of prediction error processing underlying them have a profound impact on our understanding of reinforcement-related behavior in humans, including monetary rewards and the mnemonic consequences of traumatic experiences, and for pathologies of the dopamine reinforcement system. Educational: This project will provide interdisciplinary training for postdoctoral researchers, Ph.D. and undergraduate students. The PIs will act as co-supervisors or mentors of students working in the different labs via face-to-face and internet-based technologies. We will also work with ASU’s award-winning Ask- A-Biologist program. This is an online science program designed to enrich the learning experiences of students of all ages and to provide classroom material for use by K-12 teachers. We will develop an extension of a game developed under a prior NSF award, and the new game will include modules to teach K-12 students about how insects learn. We will also integrate into the AAB site a program developed by a collaborator (B Gerber) at the Leibniz Institut für Neurobiologie, Magdeburg, and now in use in schools in Germany, to teach K-12 students how to train animals using the fruit fly larval learning paradigm. Underrepresented groups: All PIs will work with their university offices of Academic Diversity and Equal Opportunity for reaching underrepresented students.

59 BASIC BIOLOGICAL SCIENCES↗

Anion exchange membrane test protocol validation

This study presents the validation of protocols for measuring ion exchange capacity (IEC) and alkaline stability of anion exchange membranes (AEMs) for low-temperature water electrolysis. While protocols are often tested within individual laboratories, their results across multiple laboratories with varying equipment, environmental conditions, and personnel qualification remain unverified. The validation involved Los Alamos National Laboratory (LANL), National Renewable Energy Laboratory (NREL), and University of Oregon (UO) using the same commercially available AEM to assess reproducibility and reliability of the protocols under diverse conditions. For the IEC protocol, results across laboratories were consistent within ±10% of the NMR-determined reference value. The alkaline stability protocol could pose greater challenges due to factors such as variations in sample collection timing, preservation methods, and analytical techniques, but consistent test results for percentage IEC loss were demonstrated across institutions. These results highlight the reliability and applicability of the protocols, emphasizing the importance of validation to ensure consistency in diverse research environments.

08 HYDROGEN↗

Commercial Building Energy Code Field Study: Data Collection Methodology and Protocol

In support of the U.S. Department of Energy’s Commercial Buildings Energy Code Field Study, this data collection methodology and protocol provides guidance on all aspects of undertaking a compliance study, from development of a sampling plan to recruitment to code requirements and compliance checks for each energy code measure specified to be collected. The protocol also includes a data collection form that captures all key information needed for analysis of commercial energy code compliance. This methodology was developed by the Institute for Market Transformation in coordination with Pacific Northwest National Laboratory (PNNL) and the U.S. Department of Energy Building Energy Codes Program with the objective of assisting states, jurisdictions, utilities and others as they seek to measure and demonstrate compliance rates with energy codes in commercial buildings, as well as to target areas for improvement through increased energy code compliance and broader energy-efficiency programs. It is also intended to facilitate a consistent and replicable approach to research studies of this type and establish a transparent data set representing baseline construction practices across the U.S.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Commercial Building Energy Code Field Study: Data Collection Methodology and Protocol

In support of the U.S. Department of Energy’s Commercial Buildings Energy Code Field Study, this data collection methodology and protocol provides guidance on all aspects of undertaking a compliance study, from development of a sampling plan to recruitment to code requirements and compliance checks for each energy code measure specified to be collected. The protocol also includes a data collection form that captures all key information needed for analysis of commercial energy code compliance. This methodology was developed by the Institute for Market Transformation in coordination with Pacific Northwest National Laboratory (PNNL) and the U.S. Department of Energy Building Energy Codes Program with the objective of assisting states, jurisdictions, utilities and others as they seek to measure and demonstrate compliance rates with energy codes in commercial buildings, as well as to target areas for improvement through increased energy code compliance and broader energy-efficiency programs. It is also intended to facilitate a consistent and replicable approach to research studies of this type and establish a transparent data set representing baseline construction practices across the U.S.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Tuning the spin dynamics and magnetic phase transitions of the Cantor alloy via composition and sample processing protocols: A muon spin relaxation study

CrMnFeCoNi, also called the Cantor alloy, is a well-known high-entropy alloy whose magnetic properties have recently become a focus of attention. Here, we present a detailed muon spin relaxation study of the influence of chemical composition and sample processing protocols on the magnetic phase transitions and spin dynamics of several different Cantor alloy samples. Specific samples studied include a pristine equiatomic sample, samples with deficient and excess Mn content, and equiatomic samples magnetized in a field of 9 T or plastically deformed in pressures up to 0.5 GPa. The results confirm the sensitive dependence of the transition temperature on composition and demonstrate that post-synthesis pressure treatments cause the transition to become significantly less homogeneous throughout the sample volume. In addition, we observe critical spin dynamics in the vicinity of the transition in all samples, reminiscent of canonical spin glasses and magnetic materials with ideal continuous phase transitions. Application of an external magnetic field suppresses the critical dynamics in the Mn-deficient sample, while the equiatomic and Mn-rich samples show more robust critical dynamics. The spin-flip thermal activation energy in the paramagnetic phase increases with Mn content, ranging from 3.1⁢(3) × 10 -21 J for 0% Mn to 1.2⁢(2) × 10 -20 J for 30% Mn content. These results shed light on critical magnetic behavior in environments of extreme chemical disorder and demonstrate the tunability of spin dynamics in the Cantor alloy via chemical composition and sample processing.

36 MATERIALS SCIENCE↗

Predicting nepheline precipitation in waste glasses using ternary submixture model and machine learning

Nepheline precipitation in nuclear waste glasses during vitrification can be detrimental due to its negative effect on chemical durability. Developing models to accurately predict nepheline precipitation from compositions is important to increase waste loading since existing models can be overly conservative. In this study, an expanded dataset containing 955 glasses was compiled from literature data, where 355 glasses are for high-level waste (HLW). Previously developed submixture models were refitted using the new dataset, where a misclassification rate of 7.8% was achieved. Nine machine learning (ML) algorithms (e.g., k-nearest neighbor, Gaussian process regression, artificial neural network, support vector machine, decision tree, etc.) were applied to evaluate their ability of predicting nepheline precipitation from compositions. Model accuracy, precision, recall/sensitivity, and F1 score were systemically compared between different ML algorithms and modeling protocols. Good model prediction with an accuracy ~0.9 (misclassification rate of ~10%) was observed with different algorithms under certain protocol. This study evaluated various ML models to predict nepheline precipitations in waste glasses, highlighting the importance of data preparation, modeling protocol, and their effect on model stability and reproducibility. The results provide insights into applying ML to predict glass properties and suggest areas for future research on modeling nepheline precipitations.

Lu, Xiaonan↗

High-Resolution Tandem Mass Spectrometry-Based Analysis of Model Lignin–Iron Complexes: Novel Pipeline and Complex Structures

Understanding the chemical nature of soil organic carbon (SOC) with great potential to bind iron (Fe) minerals is critical for predicting the stability of SOC. Organic ligands of Fe are among the top candidates for SOCs able to strongly sorb on Fe minerals, but most of them are still molecularly uncharacterized. To shed insights into the chemical nature of organic ligands in soil and their fate, this study developed a protocol for identifying organic ligands using ultrahigh-performance liquid chromatography-high-resolution tandem mass spectrometry (UHPLC-HRMS/MS) and metabolomic tools. The protocol was used for investigating the Fe complexes formed by model compounds of lignin-derived organic ligands, namely, caffeic acid (CA), p-coumaric acid (CMA), vanillin (VNL), and cinnamic acid (CNA). Isotopologue analysis of 54/56 Fe was used to screen out the potential UHPLC-HRMS (m/z) features for complexes formed between organic ligands and Fe, with multiple features captured for CA, CMA, VNL, and CNA when 35/37 Cl isotopologue analysis was used as supplementary evidence for the complexes with Cl. MS/MS spectra, fragment analysis, and structure prediction with SIRIUS were used to annotate the structures of mono/bidentate mono/biligand complexes. The analysis determined the structures of monodentate and bidentate complexes of FeL x Cl y (L: organic ligand, x = 1–4, y = 0–3) formed by model compounds. The protocol developed in this study can be used to identify unknown organic ligands occurring in complex environmental samples and shed light on the molecular-level processes governing the stability of the SOC.

54 ENVIRONMENTAL SCIENCES↗

Effect of Radiotherapy Techniques (IMRT vs. 3D-CRT) on Outcome in Patients With Intermediate-Risk Rhabdomyosarcoma Enrolled in COG D9803-A Report From the Children's Oncology Group

Purpose: To compare the dosimetric parameters of intensity-modulated radiotherapy (IMRT) and three-dimensional conformal radiotherapy (3D-CRT) in patients with intermediate-risk rhabdomyosarcoma and to analyze their effect on locoregional control and failure-free survival (FFS). Methods and Materials: The study population consisted of 375 patients enrolled in the Children's Oncology Group protocol D9803 study, receiving IMRT or 3D-CRT. Dosimetric data were collected from 179 patients with an available composite plan. The chi-square test or Fisher's exact test was used to compare the patient characteristics and radiotherapy parameters between the two groups. The interval-to-event outcomes were estimated using the Kaplan-Meier method and compared using log-rank tests. Cox proportional hazards regression analysis was used to examine the effect of the treatment technique on FFS after adjusting for primary site and risk group. Results: The median follow-up time was 5.7 and 4.2 years for patients receiving 3D-CRT and IMRT, respectively. No differences in the 5-year failure of locoregional control (18% vs. 15%) or FFS (72% vs. 76%) rates were noted between the two groups. Multivariate analysis revealed no association between the two techniques and FFS. Patients with primary tumors in parameningeal sites were more likely to receive IMRT than 3D-CRT. IMRT became more common during the later years of the study. Patients receiving IMRT were more likely to receive >50 Gy, photon energy of {<=}6 MV, and >5 radiation fields than those who received 3D-CRT. The coverage of the IMRT planning target volume by the prescription dose was improved compared with the coverage using 3D-CRT with similar target dose heterogeneity. Conclusions: IMRT improved the target dose coverage compared with 3D-CRT, although an improvement in locoregional control or FFS could not be demonstrated in this population. Future studies comparing the integral dose to nontarget tissue and late radiation toxicity between the two groups are warranted.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Oak Ridge National Laboratory Pilot Demonstration of an Attestation and Anomaly Detection Framework using Distributed Ledger Technology for Power Grid Infrastructure

This report summarizes the design and pilot demonstration of a framework called Grid Guard that was created to provide increased data and device trustworthiness to electric grid devices by leveraging distributed ledger technology (DLT), specifically blockchain. Grid Guard contains a combination of core cryptographic methods such as the secure hash algorithm (SHA), and asymmetric cryptography, private permissioned blockchain, baselining configuration data, consensus algorithm (Raft) and the Hyperledger Fabric (HLF) framework. The system implements a low energy, fast, and robust enhancement to system trustworthiness within and across electric grid systems such as substations, control centers and metering infrastructures. Blockchain is a distributed database structured that provides a practically unalterable (immutable) timeline of stored transactions. By relying on hashing and the Raft consensus algorithm, if an entity tries to illegitimately alter a record at one instance of the database the other ledger nodes are not altered. They work to cross-reference each other and easily locate any incorrectly added data and remove it. The bulk raw data is stored in an off-chain storage (outside of the blockchain ledger) and a hash of this baseline data is stored in the Blockchain ledger via hashing windows of time-series and configuration data, after aggregation and filtering. The bulk off-chain data repository is then considered to be trust-anchored using the hashes stored in the blockchain. To secure the electric grid testbed devices and data, device configuration baselines were compared to those baselines that had been previously stored in the ledger. Statistical baselines for device configurations, network communication patterns, and high-speed sensor data are calculated and then stored off-chain and hashes stored in the ledger. Measurements such as three-phase voltage and current, frequency, breaker status, protection scheme settings, network configuration settings (and other device configuration artifacts) and network traffic features (packet interarrival times) are compared every minute or other selected time windows. During phase 1 of the Grid Guard DLT project different DLT technologies were studies, and an assessment was performed on DLT technology vulnerabilities, uses, and key characteristics. DLT consensus protocols were studies (e.g., RAFT, named after Reliable, Replicated, Redundant, And Fault-Tolerant). Also, cryptography, public, private and permissioned or permissionless systems were assessed. Grid Guard implements a permissioned private DLT. Consensus algorithm selection and choice of DLT implementation depended heavily on the use-case. For this use-case, parameters were selected to measure performance and existing tools for assessment. Benchmarking was performed theoretically and practically. During phase 2 hashed transactions/blocks were inserted into the ledger every second. During phase 2 of the Grid Guard DLT project, a prototype framework was developed and demonstrated for attestation of critical substation devices and data using precision timing systems that use PTP and IRIG-B protocols) on a testbed of operational devices that emulated a distribution substation, control center, and power metering infrastructure using real Operational Technology (OT). The testbed includes OT devices such as protective relays, human machine interfaces (HMI), and power meters. To determine when to collect and compare system and network baselines, an initial examination of an anomaly detection capability to identify malicious manipulation of data streams was conducted. The resulting anomaly detection was demonstrated in a set of experiments and leveraged to trigger device artifact attestation checks. Attestation checks occur against device configuration baselines when compared with the immutable blockchain-stored baselines, which provided a cryptographically supported means by which to store baselines. The electrical substation-grid testbed was created to test the Grid Guard framework. The testbed emulates the operations of a portion of a power grid and SCADA systems as closely as possible. The testbed integrates real protocols, mainly IEC 61850 standard protocols, such as the Sampled Value (SV) and the GOOSE protocols. The testbed also supports DNP3 and other layer 2 and layer 3 protocols such as Telnet, SSH, SFTP/FTP and other proprietary protocols needed to connect to industrial control system equipment. The testbed emulates real power conditions using the OpalRT hardware-in-the-loop (HIL) device which can create fault situations that cannot be easily tested on real systems. The electrical substation-grid testbed was created using real measurement, communication, and protection devices that electrical utilities commonly use.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Concurrent Chemoradiotherapy Followed by Consolidation Chemotherapy With Bi-Weekly Docetaxel and Carboplatin for Stage III Unresectable, Non-Small-Cell Lung Cancer: Clinical Application of a Protocol Used in a Previous Phase II Study

Purpose: To assess the clinical applicability of a protocol evaluated in a previously reported phase II study of concurrent chemoradiotherapy followed by consolidation chemotherapy with bi-weekly docetaxel and carboplatin in patients with stage III, unresectable, non-small-cell lung cancer (NSCLC). Methods and Materials: Between January 2000 and March 2006, 116 previously untreated patients with histologically proven, stage III NSCLC were treated with concurrent chemoradiotherapy. Radiation therapy was administered in 2-Gy daily fractions to a total dose of 60 Gy in combination with docetaxel, 30 mg/m{sup 2}, and carboplatin at an area under the curve value of 3 every 2 weeks during and after radiation therapy. Results: The median survival time for the entire group was 25.5 months. The actuarial 2-year and 5-year overall survival rates were 53% and 31%, respectively. The 3-year cause-specific survival rate was 60% in patients with stage IIIA disease, whereas it was 35% in patients with stage IIIB disease (p = 0.007). The actuarial 2-year and 5-year local control rates were 62% and 55%, respectively. Acute hematologic toxicities of Grade {>=}3 severity were observed in 20.7% of patients, while radiation pneumonitis and esophagitis of Grade {>=}3 severity were observed in 2.6% and 1.7% of patients, respectively. Conclusions: The feasibility of the protocol used in the previous phase II study was reconfirmed in this series, and excellent treatment results were achieved.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Commercial Building Energy Code Field Study (Final Report)

What is the dollar value of increasing compliance with the energy code? Ultimately, this is the question that policy makers, funders, and program implementers care about. With this in mind, Pacific Northwest National Laboratory (PNNL) developed a new methodology capable of determining how much energy cost savings could potentially be gained through better compliance with the code (Rosenberg et al. 2016). On March 2, 2016 DOE issued Funding Opportunity Announcement (FOA) Number DE-FOA-0001532, requesting proposals to develop and test a methodology for identifying the commercial energy code requirements that can provide the maximum cost-effective energy and cost savings through increased compliance. Awarded to the Institute for Market Transformation, this report summarizes the development and testing of a commercial code compliance methodology, including a summary of results, statistical analysis of sample, and best practices for others implementing a commercial energy code compliance field study.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Correlating the Morphological Changes to Electrochemical Performance During Carbon Corrosion in Polymer Electrolyte Fuel Cells

A mechanistic understanding of carbon corrosion in polymer electrolyte fuel cells (PEFCs) is required to design durable catalyst layers. Uncontrolled startup and shutdown of PEFCs cause electrochemical oxidation of carbon, which leads to several degradation phenomena, such as loss in electrochemical surface area (ECSA), pore structure collapse or increase in mass transport resistance. In this study, the chronology of morphological changes in the cathode catalyst layer due to carbon corrosion was identified and correlated with electrochemical performance degradation. PEFCs were subjected to the Department of Energy carbon corrosion accelerated stress test (AST) protocol. The study revealed two phases: in the initial phase (~500 AST cycles), amorphous carbon in contact with Pt nanoparticles oxidized fast. Rapid carbon loss and catalyst layer thinning occurred, but pore structure did not change significantly. Pt nanoparticles detached from the support and ECSA decreased drastically. In the second phase (~1500 AST cycles), carbon corrosion slowed down, but severe pore structure collapse was observed. Porosity and pore connectivity within the cathode catalyst layer decreased considerably. Electrochemical diagnostics corroborated this finding by showing significantly higher O2 mass transport resistance. Lastly, no significant change was observed in the concentration of oxides on the carbon surface after AST. But overall water management in the cathode catalyst layer deteriorated as the pore structure collapsed. This study provides an in-depth understanding of morphological changes during PEFC carbon corrosion AST protocol and motivates novel material design strategies to enable durable PEFCs.

catalyst layers↗

Selective Enrichment Coupled with Proteomics to Identify S-Acylated Plasma Membrane Proteins in Arabidopsis

Protein S-acylation, predominately in the form of palmitoylation, is a reversible lipid post-translational modification (PTM) on cysteines that plays important roles in protein localization, trafficking, activity, and complex assembly. The functions and regulatory mechanisms of Sacylation have been extensively studied in mammals, owing to remarkable development of the high-resolution proteomics and the discovery of the S-acylation-related enzymes. However, the advance of S-acylation studies in plants lags farther behind than that in mammals, mainly due to the lack of knowledge about proteins responsible for this process, such as protein acyltransferases (PATs) and their substrates. In this article, a systematic protocol to study global S-acylation in Arabidopsis seedlings is described. The procedures are presented in detail, including preparation of Arabidopsis seedlings, enrichment of plasma membrane (PM) proteins, followed by enrichment of S-acylated proteins/peptides based on the acyl-biotin exchange (ABE) method, and large-scale identification of S-acylated proteins/peptides via mass spectrometry. This approach enables researchers to study S-acylation of PM proteins in plants in a systematic and straightforward way. Basic protocol 1: Preparation of Arabidopsis seedling materials Basic protocol 2: Isolation and enrichment of plasma membrane proteins Support protocol 1: Determination of protein concentration using BCA assay Basic Protocol 3: Enrichment of S-acylated proteins by ABE method Support protocol 2: Protein precipitation by methanol/chloroform method Basic Protocol 4: Trypsin digestion and proteomic analysis Alternate protocol 1: Pre-resin digestion and peptide level enrichment.

S-acylation, palmitoylation, plasma membrane prote↗

SU-E-I-78: Establishing a Protocol for Quick Estimation of Thyroid Internal Contamination with 131I in Normal and Emergency Situations

Purpose: I-131 is one of the most frequent radionuclides used in nuclear medicine departments. The radiation workers, who manipulate the unsealed radio-toxic iodine, should be monitored for internal contamination. In this study a protocol was established for estimating I-131 activity absorbed in the thyroid glands of the nuclear medicine staff in normal working condition and also in accidents. Methods: I-131 with the activity of 10 μCi was injected inside the thyroid gland of a home-made anthropomorphic neck phantom. The phantom is made up of PMMA as soft tissue, and Aluminium as bone. The dose rate at different distances from the surface of the neck phantom was measured using a scintillator detector for duration of two months. Then, calibration factors were obtained, for converting the dose rate at each distance to the iodine activity inside the thyroid. Results: According to the results of this study, the calibration factors for converting the dose rates (nSv/h) at distances of 0cm, 1cm, 6cm, 11cm, and 16cm to the activity (kBq) inside the thyroid were found to be 0.03, 0.04, 0.14, 0.29, and 0.49 . Conclusion: This method can be effectively used for quick estimation of the I-131 concentration inside the thyroid of the staff for daily checks in normal working conditions and also in accidents.

07 ISOTOPE AND RADIATION SOURCES↗