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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.

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At least 991 records · Page 55
All sourcesNASA NTRSDOE OSTI
DOE OSTI2014

Energy-Performance-Based Design-Build Process: Strategies for Procuring High-Performance Buildings on Typical Construction Budgets: Preprint

NREL experienced a significant increase in employees and facilities on our 327-acre main campus in Golden, Colorado over the past five years. To support this growth, researchers developed and demonstrated a new building acquisition method that successfully integrates energy efficiency requirements into the design-build requests for proposals and contracts. We piloted this energy performance based design-build process with our first new construction project in 2008. We have since replicated and evolved the process for large office buildings, a smart grid research laboratory, a supercomputer, a parking structure, and a cafeteria. Each project incorporated aggressive efficiency strategies using contractual energy use requirements in the design-build contracts, all on typical construction budgets. We have found that when energy efficiency is a core project requirement as defined at the beginning of a project, innovative design-build teams can integrate the most cost effective and high performance efficiency strategies on typical construction budgets. When the design-build contract includes measurable energy requirements and is set up to incentivize design-build teams to focus on achieving high performance in actual operations, owners can now expect their facilities to perform. As NREL completed the new construction in 2013, we have documented our best practices in training materials and a how-to guide so that other owners and owner's representatives can replicate our successes and learn from our experiences in attaining market viable, world-class energy performance in the built environment.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗
DOE OSTI2015

Pyrococcus furiosus strains and methods of using same

Provided herein are methods for transforming a Pyrococcus furiosus with a polynucleotide. In one embodiment, the method includes contacting a P. furiosus with a polynucleotide under conditions suitable for uptake of the polynucleotide by the P. furiosus, and identifying transformants at a frequency of, for instance, at least 10.sup.3 transformants per microgram DNA. Also provided are isolated Pyrococcus furiosus having the characteristics of Pyrococcus furiosus COM1, and plasmids that include an origin of replication that functions in a Pyrococcus furiosus. The plasmid is stable in a recipient P. furiosus without selection for more than 100 generations and is structurally unchanged after replication in P. furiosus for more than 100 generations.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2015

Particle Communication and Domain Neighbor Coupling: Scalable Domain Decomposed Algorithms for Monte Carlo Particle Transport

In order to run Monte Carlo particle transport calculations on new supercomputers with hundreds of thousands or millions of processors, care must be taken to implement scalable algorithms. This means that the algorithms must continue to perform well as the processor count increases. In this paper, we examine the scalability of:(1) globally resolving the particle locations on the correct processor, (2) deciding that particle streaming communication has finished, and (3) efficiently coupling neighbor domains together with different replication levels. We have run domain decomposed Monte Carlo particle transport on up to 2 21 = 2,097,152 MPI processes on the IBM BG/Q Sequoia supercomputer and observed scalable results that agree with our theoretical predictions. These calculations were carefully constructed to have the same amount of work on every processor, i.e. the calculation is already load balanced. We also examine load imbalanced calculations where each domain’s replication level is proportional to its particle workload. In this case we show how to efficiently couple together adjacent domains to maintain within workgroup load balance and minimize memory usage.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗
DOE OSTI2016

Rapid and highly fieldable viral diagnostic

The present invention relates to a rapid, highly fieldable, nearly reagentless diagnostic to identify active RNA viral replication in a live, infected cells, and more particularly in leukocytes and tissue samples (including biopsies and nasal swabs) using an array of a plurality of vertically-aligned nanostructures that impale the cells and introduce a DNA reporter construct that is expressed and amplified in the presence of active viral replication.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

Method and apparatus for reducing memory access latency

Logic such as a memory controller writes primary data from an incoming write request as well as corresponding replicated primary data (which is a copy of the primary data) to one or more different memory banks of random access memory in response to determining a memory access contention condition for the address (including a range of addresses) corresponding to the incoming write request. When the memory bank containing the primary data is busy servicing a write request, such as to another row of memory in the bank, a read request for the primary data is serviced by reading the replicated primary data from the different memory bank of the random access memory to service the incoming read request.

97 MATHEMATICS AND COMPUTING↗
DOE OSTI2019

Investigating YAP’s role in Human Mammary Epithelial Cell (HMEC) Immortalization and Cancer Stem Cell (CSC) Phenotype Acquisition during Early Breast Cancer Progression

The malignant aspect of tumorigenesis in breast cancer is initiated when human mammary epithelial cells (HMEC) first immortalize (gain the ability to proliferate indefinitely) and subsequently undergo oncogenic activation. Our laboratory has developed a two-step HMEC immortalization process that we believe accurately models early breast cancer progression. Normal finite pre-stasis HMEC must overcome two distinct senescence barriers to achieve full immortality: stasis (stress-associated senescence) and replicative senescence, respectively. HMEC bypass stasis, and become post-stasis, when errors in the retinoblastoma pathway are present. Replicative senescence, a product of continuously shortening telomeres producing telomere dysfunction, is overcome by the reactivation of telomerase. Our lab has called HMEC that acquire the error allowing telomerase expression conditionally immortal (CI); CI cells may still have low telomerase expression. Full immortality is achieved when sufficient telomerase expression exists to maintain short stable telomeres following a process we have called conversion. The transcriptional coactivator YAP, upregulated in several human cancers, is closely correlated with a variety of oncogenic effects including proliferation and induction of cancer stem cell (CSC) properties. Our preliminary data have shown that YAP-transduced post-stasis HMEC expressed properties of CI cells. Growth curves indicated that some cell lines achieved full immortality; however, these cells had acquired an additional error – amplified expression of c-Myc, an oncogene known to induce telomerase in post-stasis or CI cells. The absence of uniform immortalization in the YAP-transduced cells suggests that YAP causes post-stasis HMEC to become conditionally immortal. This is further supported given that YAP-transduced post-stasis HMEC have decreased levels of the lncRNA MORT and rapidly go through conversion when p53 function is abrogated, well-documented attributes of CI HMEC. Previous studies have also reported that YAP controls the differentiation of breast progenitor cells; as a result, it is likely that YAP plays a role in inducing CSC phenotypes. In this study, we attempt to gain a greater understanding of YAP’s role in HMEC immortalization and induction of CSC properties in early breast cancer progression. We measured gene expression levels and attempted to identify proteins indicative of a CSC phenotype in YAP-transduced post-stasis HMEC. Our initial results did not reveal an association between YAP and HMEC conditional immortality or induction of CSC attributes in the cultures examined. However, using a YAP construct that promotes nuclear localization (where YAP is most active) has led to a potentially promising YAP-derived cell line that we presume to be immortal. We go on to demonstrate that YAP may be acting via non-transcriptional oncogenic roles in our HMEC specimens. Thus, we recommend future studies into YAP and the HMEC immortalization process given that they are promising targets for breast cancer prevention therapies.

Shah, Manav↗
DOE OSTI2020

Development and Validation of a Simulation Testbed for the Intelligent Building Agents Laboratory (IBAL) Using TRNSYS

This paper documents the development and validation of a dynamic primary cooling and thermal storage system simulation testbed. The system simulation testbed, sIBAL, is based on the Intelligent Building Agents Laboratory (IBAL) at the National Institute of Standards and Technology (NIST), which is a research infrastructure and testbed for the development, evaluation, and demonstration of intelligent control algorithms. The sIBAL testbed developed in this project will serve as a virtual twin of the real facility for future control algorithm development. The details of the methodologies used to develop and validate the simulation testbed, which replicates the dynamic behaviors of the primary cooling and ice storage system in the IBAL facility, are presented. The simulation testbed was developed in TRNSYS using built-in component models and MATLAB functions to replicate the two water-cooled chillers, a thermal storage tank, pumps, valves and other components for four different operation modes. Experiments on IBAL components were designed and executed to generate experimental data for model development and verification of the simulation platform. The validation of the simulation results was carried out in two phases: 1) independent component simulations for the chillers and thermal storage tank, and 2) a combined testbed simulation of the entire hydronic system. Comparison of simulation results to the experimental data obtained from the IBAL facility showed errors within 1 °C for the temperatures outputs of both the chiller and the thermal storage model. The error is within an acceptable range for further intelligent control algorithms development. The findings from the study are summarized and presented along with areas where additional research is needed. In addition, data filtering procedures and model refinement measures utilized to improve the accuracy and accelerate the computation time of the simulation are presented.

cooling systems↗
DOE OSTI2022

GANISP: A GAN-Assisted Importance Splitting Probability Estimator

Designing manufacturing processes with high yield and strong reliability relies on effective methods for rare event estimation. Genealogical importance splitting reduces the variance of rare event probability estimators by iteratively selecting and replicating realizations that are headed towards a rare event. The replication step is difficult when applied to deterministic systems where the initial conditions of the offspring realizations need to be modified. Typically, a random perturbation is applied to the offspring to differentiate their trajectory from the parent realization. However, this random perturbation strategy may be effective for some systems while failing for others, preventing variance reduction in the probability estimate. This work seeks to address this limitation using a generative model such as a Generative Adversarial Network (GAN) to generate perturbations that are consistent with the attractor of the dynamical system. The proposed GAN-assisted Importance SPlitting method (GANISP) improves the variance reduction for the system targeted. An implementation of the method is available in a companion repository (https://github.com/NREL/GANISP).

generative adversarial network↗
DOE OSTI2022

GANISP: A GAN-Assisted Importance Splitting Probability Estimator: Preprint

To reduce the variance of rare event probability estimator, genealogical importance splitting marches towards a rare event by iteratively selecting and replicating realizations that are headed towards a rare event. The replication step is made difficult when applied to deterministic systems as the initial conditions of the offspring realizations need to be modified. Typically, a random perturbation is applied to the offspring to differentiate their trajectory from the parent realization. It is shown that a random perturbation strategy may be effective for some systems but may also fail for others, thereby preventing variance reduction in the probability estimate. To address this limitation, it is proposed to use a generative model such as a Generative Adversarial Network (GAN) to generate perturbations that are consistent with the attractor of the dynamical system. The GAN-assisted Importance SPlitting method (GANISP) improves the variance reduction for the system targeted. An implementation of the method is available in a companion repository (https://github.com/NREL/GANISP).

generative adversarial network↗
DOE OSTI2022

If One GEB is Good, a Community of GEBs is Better

Energy efficient, connected, grid-interactive, smart and flexible buildings are key to decarbonization, lowering energy use and improving the nation’s electricity grid. The U.S. Department of Energy’s Connected Communities initiative works to demonstrate how coordinated groups of highly efficient buildings combined with other distributed energy resources (DERs), such as electric vehicle (EV) charging, batteries, storage, demand response and photovoltaic (PV) generation can reliably and cost-effectively serve as grid assets by strategically deploying efficiency and demand flexibility while reducing carbon emissions. In 2021, DOE competitively awarded $61 million to a diverse portfolio of 10 pilot projects to promote grid-interactive efficient buildings (GEBs) working together to reliably and cost-effectively serve as grid assets while decarbonizing. Two of the main tenets of the program are measuring the communities’ energy and carbon performance and understanding how to replicate project successes in other communities. This paper begins with a discussion of what Connected Communities are (including a brief history) and their many benefits, including reduced carbon emissions and increased building efficiency and demand flexibility. Next, it provides an overview of the 10 projects, highlighting the diversity of approaches to measure success and replicate the projects: geographic locations; building types; utility, regulatory, market environments; and building vintages that will be used to test the ability of buildings to serve as grid resources. It concludes with a discussion of anticipated project impacts and the metrics that will be used to evaluate the Connected Communities projects.

Nemtzow, David↗
DOE OSTI2023

Episomal DNA vectors for plant genetic engineering

This disclosure pertains to a novel platform for genetic engineering of chloroplasts. The disclosure provides episomal DNA vectors containing a chloroplast origin of replication. These vectors remain extra-plastomic and sustainably and autonomously replicate in chloroplasts of the plant cells transformed with the vectors and in the plants regenerated from the transformed plant cells. The episomal DNA vectors do not contain any sequence that shares sequence homology with the plastome DNA and, thus, do not get integrated into the plastome DNA. The vectors can also comprise one or more genes of interest that confer desirable characteristics to the transformed plant cells. The disclosure also provides methods of transforming plant cells with the episomal DNA vectors and regenerating from the transformed plant cells plants having desirable characteristics. The vectors and methods disclosed herein provide a significant advancement in speed, flexibility, and prospects of introducing genes into plant cells for effective metabolic engineering.

Lenaghan, Scott C.↗
DOE OSTI2013

Pipeline Decommissioning Trial AWE Berkshire UK - 13619

This Paper details the implementation of a 'Decommissioning Trial' to assess the feasibility of decommissioning the redundant pipeline operated by AWE located in Berkshire UK. The paper also presents the tool box of decommissioning techniques that were developed during the decommissioning trial. Constructed in the 1950's and operated until 2005, AWE used a pipeline for the authorised discharge of treated effluent. Now redundant, the pipeline is under a care and surveillance regime awaiting decommissioning. The pipeline is some 18.5 km in length and extends from AWE site to the River Thames. Along its route the pipeline passes along and under several major roads, railway lines and rivers as well as travelling through woodland, agricultural land and residential areas. Currently under care and surveillance AWE is considering a number of options for decommissioning the pipeline. One option is to remove the pipeline. In order to assist option evaluation and assess the feasibility of removing the pipeline a decommissioning trial was undertaken and sections of the pipeline were removed within the AWE site. The objectives of the decommissioning trial were to: - Demonstrate to stakeholders that the pipeline can be removed safely, securely and cleanly - Develop a 'tool box' of methods that could be deployed to remove the pipeline - Replicate the conditions and environments encountered along the route of the pipeline The onsite trial was also designed to replicate the physical prevailing conditions and constraints encountered along the remainder of its route i.e. working along a narrow corridor, working in close proximity to roads, working in proximity to above ground and underground services (e.g. Gas, Water, Electricity). By undertaking the decommissioning trial AWE have successfully demonstrated the pipeline can be decommissioned in a safe, secure and clean manor and have developed a tool box of decommissioning techniques. The tool box of includes; - Hot tapping - a method of breaching the pipe while maintaining containment to remove residual liquids, - Crimp and shear - remote crimping, cutting and handling of pipe using the excavator - Pipe jacking - a way of removing pipes avoiding excavations and causing minimal disturbance and disruption. The details of the decommissioning trial design, the techniques employed, their application and effectiveness are discussed and evaluated here in. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2015

Use of XRF to characterize pre-Hanford orchards in the 100-OL-1 operable unit - 15717

Prior to 1943, Hanford site included several small towns with approximately 5,000 acres of orchards; at the time lead arsenate (PbHAsO{sub 4}) was the most commonly used pesticide. High concentration of lead and arsenic have been recorded where trees and stumps are still visible. Pre-Hanford orchards have been designated as 100-OL-1 Operable Unit. An Optimization study evaluated optimal counting time and position of the XRF from soils collected at the site. A Pilot Study included the evaluation of a field portable x-ray fluorescence (XRF) analyzer determining if performance results meet quality assurance criteria for cleanup. XRF has only previously been used as a screening tool. Four Decision Units (DUs) are to be analyzed: Ol-14. OL-32. OL-IU6-4 and OL-FR2-1. The DUs vary in size and have had distinct historic activities; such as 46.4 acres OL-14 having a military camp in the 1950's unlike OL-FR2-1 having been disturbed as it is near the F area reactor. Together the DUs provide an adequate representation of the Operable Unit. Screening criteria for lead (250 mg/kg) and arsenic (20 mg/kg). Six samples were collected ranging above, at, and below the screening criteria of 250 mg/kg for Pb and 20 mg/kg for As at the Decision Units OL-14 and OL-IU6-4, and labeled respectively. Samples were hand sieved, homogenized and placed in sample cups. Three replicates were prepared for each sample. Each sample cup was analyzed 3 times for 30 sec. A Standard Reference Material (SRM) was analyzed after every 20 readings. Three samples were selected for 15, 30, 45, 60, 90, 120, 150 and 180 second fixed and variable analysis. Two transects at OL-14, and one at OL-IU6-4 were created by selecting locations 90. around ground zero at distances of 0.5, 1, 2, 4, 8 and 12 feet (16 ft for OL-14). Locations were scanned in triplicate for 60 seconds each. 4 Decision Units spanning roughly 370 acres were chosen to represent 100-OL-1 Operable Unit. 40 locations within each DU were selected using Visual Sample Plan (VSP) with a random start and systematic grid pattern. Each location would be analyzed with 3 replicates and a 60 second count time. The Standard Reference Material (SRM) and blank are analyzed after every 20 counts to ensure accuracy. Data is extracted from XRF to be implemented in GIS for visual representation of site with 1943 and 2013 aerial imagery. Discussion/Conclusion: - XRF provided a good indication of precision and accuracy relative to ICP with a linear correlation R{sup 2} value higher than anticipated at .9735; - MDL proved the detection limit is significantly lower than the screening criteria; - When comparing results from the first 40 locations of OL-14 to the total 80 locations of OL-14, the same variability was seen; - We expect high concentrations of Pb and As to be where past orchards are present though this does not occur as all sites. Significance: - XRF is competent for decision making as values fall within ±20% ICP data; - The highest concentrations are not where expected, which may be an issue for sites outside of Hanford that have had more development; - The size of the DU does not affect the number of samples measured. Future Work: - Verification and validation of the XRF Data (QA/QC); - Revise conceptual site model; - Statistical evaluations to investigate: Estimation of variability in surface soils, appropriate size for DU, and sample size per DU; - Coordinating sample results through CORE; - Displaying sample results in PHOENIX (GIS tool)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2018

Modeling the Drying of a Mock Used Fuel Assembly with COBRA-SFS Cycle-4 - 18597

The processing and storage of nuclear fuel is an ever increasingly pressing issue. Around 70,000 tons of used fuel are currently in either dry or wet storage on site at nuclear power plants or at specified spent fuel storage facilities. Additionally, about 2,000 tons are added to that sum each year. To promote retrievability, limit degradation in the canister, and reduce potential for hydrogen generation, spent fuel that is initially in wet storage must be dried before moving to dry cask storage. Therefore, this research seeks to use a mock used fuel storage cask to replicate the conditions in the industry during the drying process of fuel being moved from storage pools. To simulate this, a mock storage cask has been constructed to closely resemble those used in industry such that any areas that tend to retain water are replicated effectively. It is able to hold one mock used fuel assembly. The assembly used is an AREVA 10A 10 x 10 assembly with 78 rods filled with depleted uranium, 12 heater rods to simulate decay heat, a 3 x 3 BWR water channel, and one interchangeable rod position for tests such as a simulated failed fuel rod, BWR water rod, and PWR guide tube tests. The industry standard procedure is used in drying the assembly which involves stepping the system pressure down and holding for certain periods until reaching 3 torr and holding for 30 minutes. This is believed to sufficiently dry the assembly. The goal is to record data throughout these tests that can provide the industry with a greater understanding of how well the process works. This data will also be used to develop and validate a computer model of the drying process. Mechanistic models are currently under development. Reduced order models are being investigated for incorporation into current related engineering codes such as COBRA-SFS. COBRA-SFS is a member of the COBRA family of codes which is mainly used in the evaluation of thermal hydraulic systems containing spent nuclear fuel. However, it does not currently have the capability to account for processes such as evaporation encountered during drying operations. Therefore, those physical models are to be added to the source code. At present a model of the experiment has been created using the COBRA-SFS Cycle-4 code. A number of steady state models along with transient models have been developed and presented. Each of these yield valuable results for comparison with the experiments conducted. In addition to comparison with experimental results, the data collected from COBRA-SFS is compared with that which has been generated from other computer models which utilize computational fluid dynamics. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2019

Oxide Growth and Validation for Aluminum Research Reactor Fuel - 19422

Aluminum fuel cladding will undergo general corrosion with resulting formation of adherent oxide films during in-reactor and post-discharge exposures under various water conditions and temperatures. These oxides include chemically-bound water that can pose challenges for extended dry storage. This paper describes laboratory experiments performed to produce hydroxide (oxide) films on aluminum 1100, 6061 and 5052 series alloy substrates immersed in water at controlled temperatures. The results are compared to the oxide film present on a spent fuel assembly irradiated in research reactor. This work is part of a broad investigation to address knowledge gaps and technical data needs for dry storage of aluminum-clad spent nuclear fuel (SNF). To adequately assess the risks posed by chemically-bound water on SNF in dry storage and strategies for removing chemically bound water, it is necessary to understand the expected composition and loading of oxide films on SNF of different service histories and to investigate the response of such oxides to irradiation and potential drying methods. These efforts require both the growth of films under controlled conditions and the production of non-radioactive surrogates for testing. The current work comprises experimental investigation of oxide growth via (1) laboratory experiments to produce hydroxide films on aluminum alloy substrates immersed in water at controlled temperatures and (2) characterization and comparison of laboratory-grown oxides to oxides on service-exposed aluminum alloys. The important factors affecting oxide growth and characteristics typical of oxide films formed by aqueous corrosion under conditions similar to in-reactor are reviewed. Aluminum oxide films were grown under a variety of controlled temperature conditions and durations to serve as surrogates for oxides found on aluminum-clad spent nuclear fuel due to aqueous corrosion. The chemically-bound water found in oxides formed by aqueous corrosion poses challenges for extended dry storage, requiring investigation of how the oxide loading and characteristics vary with exposure history and to provide the initial condition for investigation of drying methods. Characteristics including chemical composition, oxide morphology, and film thickness were measured to assess whether the surrogate films were representative of in-service-produced oxides on SNF. Tests were conducted in liquid water at atmospheric pressure in the temperature range 50-100 deg. C, with and without heat flux through the corroding surface, to replicate in-service conditions, and at 185 deg. C inside pressure vessels to replicate pre-filming procedures. Three different alloys commonly used in aluminum clad research reactor fuel were considered: Al-1100, Al-6061, and Al-5052. Specimens were characterized by weighing before and after exposure to measure oxide mass loading, X-ray diffraction to identify oxide composition, and scanning electron microscopy to examine the oxide morphology and film thickness. The oxide type and the morphological characteristics of the film on a fuel element from the Uruguay RU1 research reactor were determined and compared to the laboratory surrogates. The results show some differences compared to the 50 deg. C laboratory specimen that are rationalized with the service storage history of the element. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2017

Quantitative Proteomic Analysis of Optimal Cutting Temperature (OCT) Embedded Core-Needle Biopsy of Lung Cancer

With recent advances in understanding the genomic underpinnings and oncogenic drivers of pathogenesis in different subtypes, it is increasingly clear that proper pretreatment diagnostics are essential for the choice of appropriate treatment options for non-small cell lung cancer (NSCLC). Tumor tissue preservation in optimal cutting temperature (OCT) compound is commonly used in the surgical suite. However, proteins recovered from OCT-embedded specimens pose a challenge for LC-MS/MS experiments, due to the large amounts of polymers present in OCT. Here we present a simple workflow for whole proteome analysis of OCT-embedded NSCLC tissue samples, which involves a simple trichloroacetic acid precipitation step. Comparisons of protein recovery between frozen versus OCT-embedded tissue showed excellent consistency with more than 9200 proteins identified. Using an isobaric labeling strategy, we quantified more than 5400 proteins in tumor versus normal OCT-embedded core needle biopsy samples. Gene ontology analysis indicated that a number of proliferative as well as squamous cell carcinoma (SqCC) marker proteins were overexpressed in the tumor, consistent with the patient’s pathology based diagnosis of “poorly differentiated SqCC”. Among the most downregulated proteins in the tumor sample, we noted a number of proteins with potential immunomodulatory functions. Finally, interrogation of the aberrantly expressed proteins using a candidate approach and cross-referencing with publicly available databases led to the identification of potential druggable targets in DNA replication and DNA damage repair pathways. We conclude that our approach allows LC-MS/MS proteomic analyses on OCT-embedded lung cancer specimens, opening the way to bring powerful proteomics into the clinic. .

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2012

Heat induces gene amplification in cancer cells

Highlights: Black-Right-Pointing-Pointer This study discovered that heat exposure (hyperthermia) results in gene amplification in cancer cells. Black-Right-Pointing-Pointer Hyperthermia induces DNA double strand breaks. Black-Right-Pointing-Pointer DNA double strand breaks are considered to be required for the initiation of gene amplification. Black-Right-Pointing-Pointer The underlying mechanism of heat-induced gene amplification is generation of DNA double strand breaks. -- Abstract: Background: Hyperthermia plays an important role in cancer therapy. However, as with radiation, it can cause DNA damage and therefore genetic instability. We studied whether hyperthermia can induce gene amplification in cancer cells and explored potential underlying molecular mechanisms. Materials and methods: (1) Hyperthermia: HCT116 colon cancer cells received water-submerged heating treatment at 42 or 44 Degree-Sign C for 30 min; (2) gene amplification assay using N-(phosphoacetyl)-L-aspartate (PALA) selection of cabamyl-P-synthetase, aspartate transcarbarmylase, dihydro-orotase (cad) gene amplified cells; (3) southern blotting for confirmation of increased cad gene copies in PALA-resistant cells; (4) {gamma}H2AX immunostaining to detect {gamma}H2AX foci as an indication for DNA double strand breaks. Results: (1) Heat exposure at 42 or 44 Degree-Sign C for 30 min induces gene amplification. The frequency of cad gene amplification increased by 2.8 and 6.5 folds respectively; (2) heat exposure at both 42 and 44 Degree-Sign C for 30 min induces DNA double strand breaks in HCT116 cells as shown by {gamma}H2AX immunostaining. Conclusion: This study shows that heat exposure can induce gene amplification in cancer cells, likely through the generation of DNA double strand breaks, which are believed to be required for the initiation of gene amplification. This process may be promoted by heat when cellular proteins that are responsible for checkpoints, DNA replication, DNA repair and telomere functions are denatured. To our knowledge, this is the first study to provide direct evidence of hyperthermia induced gene amplification.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2013

Phosphorylation at tyrosine 114 of Proliferating Cell Nuclear Antigen (PCNA) is required for adipogenesis in response to high fat diet

Highlights: Black-Right-Pointing-Pointer Proliferating Cell Nuclear Antigen (PCNA) is phosphorylated at Y114. Black-Right-Pointing-Pointer Phospho-Y114 of PCNA is not required for cell proliferation for normal growth. Black-Right-Pointing-Pointer MCE during adipogenesis is abolished in the lack of the phosphorylation. Black-Right-Pointing-Pointer Homozygous Y114F mice are resistant to high fat diet induced obesity. Black-Right-Pointing-Pointer Our results shed light on the interface between proliferation and differentiation. -- Abstract: Clonal proliferation is an obligatory component of adipogenesis. Although several cell cycle regulators are known to participate in the transition between pre-adipocyte proliferation and terminal adipocyte differentiation, how the core DNA synthesis machinery is coordinately regulated in adipogenesis remains elusive. PCNA (Proliferating Cell Nuclear Antigen) is an indispensable component for DNA synthesis during proliferation. Here we show that PCNA is subject to phosphorylation at the highly conserved tyrosine residue 114 (Y114). Replacing the Y114 residue with phenylalanine (Y114F), which is structurally similar to tyrosine but cannot be phosphorylated, does not affect normal animal development. However, when challenged with high fat diet, mice carrying homozygous Y114F alleles (PCNA{sup F/F}) are resistant to adipose tissue enlargement in comparison to wild-type (WT) mice. Mouse embryonic fibroblasts (MEFs) harboring WT or Y114F mutant PCNA proliferate at similar rates. However, when subjected to adipogenesis induction in culture, PCNA{sup F/F} MEFs are not able to re-enter the cell cycle and fail to form mature adipocytes, while WT MEFs undergo mitotic clonal expansion in response to the adipogenic stimulation, accompanied by enhanced Y114 phosphorylation of PCNA, and differentiate to mature adipocytes. Consistent with the function of Y114 phosphorylation in clonal proliferation in adipogenesis, fat tissues isolated from WT mice contain significantly more adipocytes than those isolated from PCNA{sup F/F} mice. This study identifies a critical role for PCNA in adipose tissue development, and for the first time identifies a role of the core DNA replication machinery at the interface between proliferation and differentiation.

60 APPLIED LIFE SCIENCES↗