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All sourcesNASA NTRSDOE OSTI
NASA NTRSDate not supplied

Dose, LET, time and strain dependence of radiation-induced 53BP1 foci in 15 mouse strains ex vivo and associations to in vivo radiation susceptibility

We present a comparative analysis on the repair of radiation-induced DNA damage ex vivo in 15 strains of mice, including 5 inbred reference strains and 10 collaborative-cross strains, of both sexes. Non-immortalized primary skin fibroblasts derived from 76 mice were subjected to both low- and high-LET radiation (0.1, 1 and 4 Gy of X rays; 1.1 and 3 particles/100μm2 of 350 MeV/n 40Ar and 600 MeV/n 56Fe). Automated image quantification of 53BP1 radiation-induced foci (RIF) during the first 4-48 h post-irradiation was performed as a function of dose and LET. Similarly to what we had previously reported for immortalized human cell lines [1], we observed a saturation of RIF number with dose at 4h post-irradiation, with more RIF/Gy for lower LET (X rays and 40Ar) compared to 56Fe. However at later time points (24h and above), the trend was inverted with more RIF/Gy for higher LET. Our data suggest that multiple DSBs cluster into RIF: as the linear density of DSBs increases with LET, so does the probability of having more DSBs per RIF, which makes it more difficult for cells to fully resolve high-LET-induced RIF, explaining the hypersensitivity to high-LET radiation despite a low number of RIF. Taking into account the amount of clustering at a given dose and LET, but also the kinetics of DNA damage repair, we introduced a novel mathematical formalism to evaluate the number of remaining RIF over time. We showed that the newly introduced kinetic metrics can be used as surrogate biomarkers for in vivo radiation toxicity, with potential applications in radiotherapy and human space exploration. In particular, we observed an association between the repairable fraction of RIF measured in vitro and survival levels of immune cells collected from irradiated mice. Moreover, the speed of DNA damage repair correlated with spontaneous cancer incidence data collected from the Mouse Tumor Biology database, suggesting a relationship between the efficiency of DSB repair after irradiation and cancer risk. In addition to the efficacy of repair and persistent RIF levels, even the amount of spontaneous foci without irradiation was shown to be strain dependent, indicating that these phenotypes are at least partially driven by genetics, and supporting their potential as indicators of individual radiation sensitivity. [1] Neumaier, T., et al., PNAS, 2012 (8) 109:443

Radiation, DNA damage, repair kinetics↗
DOE OSTI2023

Identification of recG genetic interactions in Escherichia coli by transposon sequencing

ABSTRACT Maintaining the integrity of the genome is of utmost importance for cell division and propagation. In Escherichia coli , the RecG protein has been implicated in processing branched recombination intermediates during DNA repair processes, but the primary cellular role(s) of RecG and the repair pathways in which it acts have been difficult to define. To gain additional insight into RecG function, we employed transposon sequencing to identify recG genetic interactions and reveal complementary or redundant functions. The strongest hits from the screen were the dam , uvrD , rnhA , radA , and rep genes. The conditional importance of these five genes in cells lacking recG was confirmed using a plasmid-based assay, revealing synthetic lethal interactions for most double deletion strains. Several of the synthetic lethal gene combinations (with uvrD, rep, and radA , but not rnhA or dam ) were suppressed by deletion of recF or recO , indicating that their genetic relationships involved roles in post-replication gap repair. Additionally, loss of the RecG/SSB interaction phenocopied a recG deletion when combined with dam , uvrD , radA , or rnhA deletions but not with rep . The results reinforce the idea of RecG as a general genome guardian. RecG has at least two functions. It plays an important role in the resolution of joint molecules behind the fork, formed during post-replication gap repair. RecG is also required to suppress genome over-replication caused by unscheduled replication initiation at R-loops, at double-strand breaks caused by dam inactivation, and during replication termination. IMPORTANCE DNA damage and subsequent DNA repair processes are mutagenic in nature and an important driver of evolution in prokaryotes, including antibiotic resistance development. Genetic screening approaches, such as transposon sequencing (Tn-seq), have provided important new insights into gene function and genetic relationships. Here, we employed Tn-seq to gain insight into the function of the recG gene, which renders Escherichia coli cells moderately sensitive to a variety of DNA-damaging agents when they are absent. The reported recG genetic interactions can be used in combination with future screens to aid in a more complete reconstruction of DNA repair pathways in bacteria.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

MLEC-Sim: A Simulator for Evaluating Multi-Level Erasure Coding

We present MLEC-Sim, a sophisticated simulator for Multi-Level Erasure Coding (MLEC), developed in approximately 13 KLOC. The simulator is engineered to analyze the impact of various system configurations and erasure coding policies on system durability and network overhead. It supports a comprehensive range of parameters including disk capacity, disk I/O bandwidth, failure rates, network bandwidth, and system scale, accommodating various erasure coding approaches such as Single-Level Erasure Coding (SLEC), Multi-Level Erasure Coding (MLEC), and Local Reconstruction Codes (LRC). MLEC-Sim provides support for multiple chunk placement policies, including clustered parity and declustered parity, and encompasses a variety of repair methods like Repair-ALL, Repair-FCO, Repair-HYB, and Repair-MIN. It is capable of simulating disk failures through a variety of means, including distribution-based or trace-based mechanisms, and can handle complex multi-level (de)clustered placements and repair processes. A key feature of MLEC-Sim is its adoption of the splitting simulation method for evaluating system durabilities at extremely high levels, which are challenging to assess with traditional simulation approaches. This feature allows for a detailed evaluation of system resilience under a range of conditions, aiding in the selection of appropriate erasure coding solutions for enhancing system durability. MLEC-Sim contributes to the field of data storage and reliability by providing a tool for the detailed evaluation of the durability and efficiency of erasure coding configurations, intended for use by researchers and practitioners in the design and optimization of storage systems.

Wang, Meng↗
DOE OSTI2023

Quantifying Uncertainty of Deep Reinforcement Learning Based Decision Making for Operations and Maintenance of Nuclear Power Plant

This paper summarizes research that integrates condition monitoring and prognostics with decision making for nuclear power plant operations and maintenance. As part of this research, we have developed an online asset management tool to help reduce life-cycle maintenance and repair costs. Using the latest advancements in condition monitoring, supply chain analytics, and deep reinforcement learning, we have created a predictive maintenance tool that can optimize the maintenance and spare-part management of a repairable nuclear system. To demonstrate these methods, preliminary studies were conducted on a simple, representative maintenance system undergoing a stochastic degradation process that requires repairs or replacement to continue operation. Through Monte Carlo simulations, we were able to reduce maintenance spending by approximately 50% compared to optimized, time-based maintenance strategies. Not only does the decision maker reduce the average life-cycle costs, it also minimizes the chance of high cost scenarios, lowering the variance of the expected cost distributions, and reducing overall financial risk. Furthermore, this work also studies the ability of the decision maker to handle various levels of noise from observation uncertainty. By introducing uncertainty into the decision-making process, we have quantified the robustness and resiliency of the decision maker, as well as identified necessary levels of observability to demonstrate cost effectiveness.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗
DOE OSTI2026

Nondestructive Evaluation of Carbon Fiber Reinforced Polymers

The American Society of Mechanical Engineers Boiler and Pressure Vessel Code requires repair and replacement of safety-related piping materials to meet the original Construction Code; however, no construction criteria currently exist for carbon fiber reinforced polymer (CFRP) materials in the nuclear industry. Although nondestructive examination (NDE) techniques for cast and wrought ferritic and austenitic steels are well established, licensees are increasingly deploying novel materials such as CFRP for which assessment of the use of NDE is required, as the inspectability of such materials and the influence of manufacturing processes on inspectability remain insufficiently understood. To address these gaps, Pacific Northwest National Laboratory (PNNL) evaluated the fabrication of several CFRP repair mockups and the effectiveness of NDE methods to support regulatory review of CFRP repairs in nuclear power plant applications. Representative flat-plate mockups containing realistic fabrication defects were manufactured and inspected using manual and automated tap testing, dynamic response spectroscopy (DRS), and ultrasonic testing (UT). The study identified significant fabrication variability, particularly in controlling defect size and epoxy thickness, which strongly influenced defect detectability by NDE methods. Tap testing was effective for shallow defects in thin epoxy configurations but unreliable for thicker repairs and deeper flaws. DRS demonstrated higher sensitivity to dry spots but produced unconfirmed indications for thicker plates, requiring further validation. Conventional UT, particularly at 1.0 MHz, showed the strongest overall capability for detecting a range of defects, although performance degraded with increasing repair thickness and complexity. The results highlight the need to better define critical defect sizes, develop reliable mockup fabrication methods, and validate NDE methods needed to support CFRP repairs.

36 MATERIALS SCIENCE↗
NASA NTRS1971

Protective coatings for columbium applied in the field.

The various aspects of field repair of columbium alloy panels with protective coatings designed as part of the Space Shuttle thermal protection system are examined. The field repair of the coatings is accomplished by employing ceramic cement repairs, and reapplying the fused slurry silicide coating. Techniques are described which improve the practicality of these repairs by employing torch heating. The repair coating quality is demonstrated by testing which simulates flight temperature, pressure, stress and acoustic vibration conditions as a function of time. Conclusions on the present status of field repair coatings are presented and recommendations are given for appropriate future activities relative to the use on an operational Space Shuttle system.

Carter, J.↗
NASA NTRS1991

A CLIPS based personal computer hardware diagnostic system

Often the person designated to repair personal computers has little or no knowledge of how to repair a computer. Described here is a simple expert system to aid these inexperienced repair people. The first component of the system leads the repair person through a number of simple system checks such as making sure that all cables are tight and that the dip switches are set correctly. The second component of the system assists the repair person in evaluating error codes generated by the computer. The final component of the system applies a large knowledge base to attempt to identify the component of the personal computer that is malfunctioning. We have implemented and tested our design with a full system to diagnose problems for an IBM compatible system based on the 8088 chip. In our tests, the inexperienced repair people found the system very useful in diagnosing hardware problems.

Whitson, George M.↗
NASA NTRS2011

Atomic Simulation of Complex DNA DSBs and the Interactions with the Ku70/80 Heterodimer

DNA double strand breaks (DSBs) induced by ionizing radiation (IR) usually contain modified bases such as 8-oxo-7,8-dihydroguanine (8-oxoG) and thymine glycol, apurinic/apyrimidinic (AP) sites, 2-deoxyribonolactone, or single-strand breaks (SSBs). The presence of such lesions in close proximity to the DSB terminus makes the DNA nicks more difficult to repair and rejoin than endogenously induced simple DSBs, and as such a major determinant of the biological effects of high linear energy transfer (LET) radiation as encountered in space travel. In this study we conducted molecular dynamics simulations on a series of DNA duplexes with various complex lesions of 8-oxoG and AP sites, in an effort to investigate the effects of such lesions to the structural integrity and stability of DNA after insulted by IR. We also simulated the interaction of such complex DSBs with the Ku70/80 heterodimer, the first protein in mammalian cells to embark the non-homologous end joining (NHEJ) DNA repair pathway. The results indicate, compared to DNA with simple DSBs, the complex lesions can enhance the hydrogen bonds opening rate at the DNA terminus, and increase the mobility of the whole duplex, thus they present more deleterious effects to the genome integrity if not captured and repaired promptly in cells. Simulations also demonstrate the binding of Ku drastically reduces structural disruption and flexibility caused by the complex lesions, and the interactions of Ku with complex DSBs have a different potential energy landscape from the bound structure with simple DSB. In all complex DSBs systems, the binding of DSB terminus with Ku70 is softened while the binding of the middle duplex with Ku80 is tightened. This energy shift may help the Ku protein to secure at the DSB terminus for a longer time, so that other end processing factors or repair pathways can proceed at the lesions before NHEJ repair process starts. These atomic simulations may provide valuable new insight into the selective action of repair proteins on damaged DNA.

Hu, Shaowen↗
NASA NTRS2013

Modeling Non-homologous End Joining

Non-homologous end joining (NHEJ) is the dominant DNA double strand break (DSB) repair pathway and involves several NHEJ proteins such as Ku, DNA-PKcs, XRCC4, Ligase IV and so on. Once DSBs are generated, Ku is first recruited to the DNA end, followed by other NHEJ proteins for DNA end processing and ligation. Because of the direct ligation of break ends without the need for a homologous template, NHEJ turns out to be an error-prone but efficient repair pathway. Some mechanisms have been proposed of how the efficiency of NHEJ repair is affected. The type of DNA damage is an important factor of NHEJ repair. For instance, the length of DNA fragment may determine the recruitment efficiency of NHEJ protein such as Ku [1], or the complexity of the DNA breaks [2] is accounted for the choice of NHEJ proteins and subpathway of NHEJ repair. On the other hand, the chromatin structure also plays a role of the accessibility of NHEJ protein to the DNA damage site. In this talk, some mathematical models of NHEJ, that consist of series of biochemical reactions complying with the laws of chemical reaction (e.g. mass action, etc.), will be introduced. By mathematical and numerical analysis and parameter estimation, the models are able to capture the qualitative biological features and show good agreement with experimental data. As conclusions, from the viewpoint of modeling, how the NHEJ proteins are recruited will be first discussed for connection between the classical sequential model [4] and recently proposed two-phase model [5]. Then how the NHEJ repair pathway is affected, by the length of DNA fragment [6], the complexity of DNA damage [7] and the chromatin structure [8], will be addressed

Li, Yongfeng↗
NASA NTRS2018

Coalescence of DNA Double Strand Breaks Induced by Galactic Cosmic Radiation is Modulated by Genetics in 15 Inbred Strains of Mice

In this manuscript we address the challenges associated with the ability to predict radiation sensitivity associated with exposure to either cosmic radiation or X-rays in a population study, by monitoring DNA damage sensing protein 53BP1 forming small nuclear radiation-induced foci (RIF) as a surrogate biomarker of DNA double strand breaks (DSB). 76 primary skin fibroblasts were isolated from 10 collaborative cross strains and five reference inbred mice (C57Bl/6, BALB/CByJ, B6C3, C3H and CBA/CaJ) and exposed to three different charged nuclei of increasing LET (350 MeV/n Si, 350 MeV/n Ar and 600 MeV/n Fe) and X-ray. Our data brings strong evidence against the classic "contact-first" model where DSBs are assumed to be immobile and repaired at the lesion site. In contrast, our model suggests nearby DSBs move into single repair unit characterized by large RIF before the repair machinery kicks in. Such model has the advantage of being much more efficient molecularly but is poorly suited to deal with cosmic radiation, where energy is concentrated along the particle trajectory, inducing a large density of DSBs along each particle track. In accordance with this model, RIF quantification after X-ray exposition showed a saturated dose response for early time points post-irradiation for all strains. Similarly, the high-LET response showed that RIF number matched the number of track per cell, not the number of expected DSB per cell (1). At the temporal level, we noted that the percentage of unrepaired high-LET tracks over a 48 hour time-course increased with LET, confirming that the DNA repair process becomes more difficult as more DSB coalesce into single RIF. There was also good agreement between persistent RIF levels measured in-vitro in the primary skin cultures and survival levels of T-cells and B-cells collected in blood samples from 10 CC strains 24 hours after 0.1 Gy whole-body dose of X-ray. This suggests that persistent RIF 24 hour post-IR is a good surrogate in-vitro biomarker for in-vivo radiation toxicity. Finally, at the genomic level, large differences in repair rates between strains for high-LET allowed us to identify suggestive genetic loci associated with radiation sensitivity. Interestingly, the two highest LETs provided the most strain variation with a common locus on Chromosome 10 highly enriched for DNA repair associated genes we discussed in detail.

Radiation-Induced Foci↗
DOE OSTI2024

Mitigation for Removal of Building 01-103 in the Area 1 Subdock, Nevada National Security Site, Nye County, Nevada

Building 01-103, the Drill Bit Repair Building was originally recorded as SHPO Resource No. B18848 in 2020 during an architectural survey of the Area 1 Subdock (Collins et al. 2022). It was recommended individually eligible for listing in the NRHP under Criteria A and C and contributing to the significance of the Area 1 Subdock Historic District. The SHPO concurred with these determinations in a letter dated April 29, 2022 (Reed 2022). The Drill Bit Repair Building retains all seven aspects of integrity. The location, materials, workmanship, feeling, and association have not changed since its original construction. The design was modified slightly in 1996 with the addition of a removable and non-destructive chain-link fence. Nearby buildings 01-101 and 01-102 were removed (Collins and Menocal 2020) but the other buildings and storage yards remain. Therefore, the integrity of setting has been changed, but not to the degree that the building no longer conveys its significance (see Figures 4 and 5 for comparison). Building 01-103 was constructed in 1985 to coincide with the relocation of the Area 3 Subdock to Area 1. The design for the new Subdock provided a centralized cluster of buildings surrounded by two large storage yards. The buildings included administrative and engineering office space, warehouses, a radiograph building, and Building 01-103 as the drill bit maintenance and repair building, among others. The Drill Bit Repair Building was critical to the success of underground nuclear testing and is eligible under Criterion A for its contribution to broad patterns of history by maintaining specially designed equipment for underground nuclear testing at the NNSS. It is also eligible under Criterion C as it is the only recorded example of a facility built specifically for the maintenance of big-hole drill bits on the NNSS. As it retains both significance and integrity, it remains individually eligible under Criteria A and C. In addition to its individual significance, the Drill Bit Repair Building also contributes to the significance of the Area 1 Subdock Historic District as a representative of a big-hole drilling support facility necessary for underground nuclear testing. The building and the equipment therein were designed specifically for testing at the NNSS, where much of the early technology and methods for big-hole drilling were developed. Its removal will result in not only the demolition of a contributing element but also a change in the physical features of the district’s setting that contribute to its historic significance. The change in setting will diminish the district’s integrity. However, the district still contains numerous examples of other buildings constructed for the purpose of supporting big-hole drilling and remains eligible under Criteria A and C.

54 ENVIRONMENTAL SCIENCES↗
NASA NTRS2002

Gene conversion is strongly induced in human cells by double-strand breaks and is modulated by the expression of BCL-x(L)

Homology-directed repair (HDR) of DNA double-strand breaks (DSBs) contributes to the maintenance of genomic stability in rodent cells, and it has been assumed that HDR is of similar importance in DSB repair in human cells. However, some outcomes of homologous recombination can be deleterious, suggesting that factors exist to regulate HDR. We demonstrated previously that overexpression of BCL-2 or BCL-x(L) enhanced the frequency of X-ray-induced TK1 mutations, including loss of heterozygosity events presumed to arise by mitotic recombination. The present study was designed to test whether HDR is a prominent DSB repair pathway in human cells and to determine whether ectopic expression of BCL-x(L) affects HDR. Using TK6-neo cells, we find that a single DSB in an integrated HDR reporter stimulates gene conversion 40-50-fold, demonstrating efficient DSB repair by gene conversion in human cells. Significantly, DSB-induced gene conversion events are 3-4-fold more frequent in TK6 cells that stably overexpress the antiapoptotic protein BCL-X(L). Thus, HDR plays an important role in maintaining genomic integrity in human cells, and ectopic expression of BCL-x(L) enhances HDR of DSBs. This is the first study to highlight a function for BCL-x(L) in modulating DSB repair in human cells.

NASA Discipline Radiation Health↗
NASA NTRS2000

Ionizing-radiation resistance in the desiccation-tolerant cyanobacterium Chroococcidiopsis

The effect of X-ray irradiation on cell survival, induction, and repair of DNA damage was studied by using 10 Chroococcidiopsis strains isolated from desert and hypersaline environments. After exposure to 2.5 kGy, the percentages of survival for the strains ranged from 80 to 35%. In the four most resistant strains, the levels of survival were reduced by 1 or 2 orders of magnitude after irradiation with 5 kGy; viable cells were recovered after exposure to 15 kGy but not after exposure to 20 kGy. The severe DNA damage evident after exposure to 2.5 kGy was repaired within 3 h, and the severe DNA damage evident after exposure to 5 kGy was repaired within 24 h. The increase in trichloroacetic acid-precipitable radioactivity in the culture supernatant after irradiation with 2.5 kGy might have been due to cell lysis and/or an excision process involved in DNA repair. The radiation resistance of Chroococcidiopsis strains may reflect the ability of these cyanobacteria to survive prolonged desiccation through efficient repair of the DNA damage that accumulates during dehydration.

Non-NASA Center↗
DOE OSTI2022

Cas9-induced large deletions and small indels are controlled in a convergent fashion

Repair of Cas9-induced double-stranded breaks results primarily in formation of small insertions and deletions (indels), but can also cause potentially harmful large deletions. While mechanisms leading to the creation of small indels are relatively well understood, very little is known about the origins of large deletions. Using a library of clonal NGS-validated mouse embryonic stem cells deficient for 32 DNA repair genes, we have shown that large deletion frequency increases in cells impaired for non-homologous end joining and decreases in cells deficient for the central resection gene Nbn and the microhomology-mediated end joining gene Polq. Across deficient clones, increase in large deletion frequency was closely correlated with the increase in the extent of microhomology and the size of small indels, implying a continuity of repair processes across different genomic scales. Furthermore, by targeting diverse genomic sites, we identified examples of repair processes that were highly locus-specific, discovering a role for exonuclease Trex1. Finally, we present evidence that indel sizes increase with the overall efficiency of Cas9 mutagenesis. These findings may have impact on both basic research and clinical use of CRISPR-Cas9, in particular in conjunction with repair pathway modulation.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

DNA double-strand break movement in heterochromatin depends on the histone acetyltransferase dGcn5

Abstract Cells employ diverse strategies to repair double-strand breaks (DSBs), a dangerous form of DNA damage that threatens genome integrity. Eukaryotic nuclei consist of different chromatin environments, each displaying distinct molecular and biophysical properties that can significantly influence the DSB-repair process. DSBs arising in the compact and silenced heterochromatin domains have been found to move to the heterochromatin periphery in mouse and Drosophila to prevent aberrant recombination events. However, it is poorly understood how chromatin components, such as histone post-translational modifications, contribute to these DSB movements within heterochromatin. Using irradiation as well as locus-specific DSB induction in Drosophila tissues and cultured cells, we find enrichment of histone H3 lysine 9 acetylation (H3K9ac) at DSBs in heterochromatin but not euchromatin. We find this increase is mediated by the histone acetyltransferase dGcn5, which rapidly localizes to heterochromatic DSBs. Moreover, we demonstrate that in the absence of dGcn5, heterochromatic DSBs display impaired recruitment of the SUMO E3 ligase Nse2/Qjt and fail to relocate to the heterochromatin periphery to complete repair. In summary, our results reveal a previously unidentified role for dGcn5 and H3K9ac in heterochromatic DSB repair and underscore the importance of differential chromatin responses at heterochromatic and euchromatic DSBs to promote safe repair.

Biochemistry & Molecular Biology↗
DOE OSTI2021

Genomic reconstruction of fossil and living microorganisms in ancient Siberian permafrost

Abstract Background Total DNA (intracellular, iDNA and extracellular, eDNA) from ancient permafrost records the mixed genetic repository of the past and present microbial populations through geological time. Given the exceptional preservation of eDNA under perennial frozen conditions, typical metagenomic sequencing of total DNA precludes the discrimination between fossil and living microorganisms in ancient cryogenic environments. DNA repair protocols were combined with high throughput sequencing (HTS) of separate iDNA and eDNA fraction to reconstruct metagenome-assembled genomes (MAGs) from ancient microbial DNA entrapped in Siberian coastal permafrost. Results Despite the severe DNA damage in ancient permafrost, the coupling of DNA repair and HTS resulted in a total of 52 MAGs from sediments across a chronosequence (26–120 kyr). These MAGs were compared with those derived from the same samples but without utilizing DNA repair protocols. The MAGs from the youngest stratum showed minimal DNA damage and thus likely originated from viable, active microbial species. Many MAGs from the older and deeper sediment appear related to past aerobic microbial populations that had died upon freezing. MAGs from anaerobic lineages, including Asgard archaea, however exhibited minimal DNA damage and likely represent extant living microorganisms that have become adapted to the cryogenic and anoxic environments. The integration of aspartic acid racemization modeling and metaproteomics further constrained the metabolic status of the living microbial populations. Collectively, combining DNA repair protocols with HTS unveiled the adaptive strategies of microbes to long-term survivability in ancient permafrost. Conclusions Our results indicated that coupling of DNA repair protocols with simultaneous sequencing of iDNA and eDNA fractions enabled the assembly of MAGs from past and living microorganisms in ancient permafrost. The genomic reconstruction from the past and extant microbial populations expanded our understanding about the microbial successions and biogeochemical alterations from the past paleoenvironment to the present-day frozen state. Furthermore, we provided genomic insights into long-term survival mechanisms of microorganisms under cryogenic conditions through geological time. The combined strategies in this study can be extrapolated to examine other ancient non-permafrost environments and constrain the search for past and extant extraterrestrial life in permafrost and ice deposits on Mars.

54 ENVIRONMENTAL SCIENCES↗
NASA NTRS1975

Statistical estimation of service cracks and maintenance cost for aircraft structures

A method is developed for the statistical estimation of the number of cracks to be repaired in service as well as the repair and the maintenance costs. The present approach accounts for the statistical distribution of the initial crack size, the statistical nature of the NDI technique used for detecting the crack, and the renewal process for the crack propagation of repaired cracks. The mean and the standard deviation of the cumulative number of cracks to be repaired are computed as a function of service time. The statistics of the costs of repair and maintenance, expressed in terms of the percentage of the cost of replacement, are estimated as a function of service time. The results of the present study provide relevant information for the decision of fleet management, the estimation of life cycle cost, and procurement specifications. The present study is essential to the design and cost optimization of aircraft structures.

Yang, J.-N.↗
NASA NTRS1992

Effect of spaceflight on the extracellular matrix of skeletal muscle after a crush injury

The organization and composition of the extracellular matrix were studied in the crush-injured gastrocnemius muscle of rats subjected to 0 G. After 14 days of flight on Cosmos 2044, the gastrocnemius muscle was removed and evaluated by histochemical and immunohistochemical techniques from the five injured flight rodents and various earth-based treatment groups. In general, the repair process was similar in all injured muscle samples with regard to the organization of the extracellular matrix and myofibers. Small and large myofibers were present within an expanded extracellular matrix, indicative of myogenesis and muscle regeneration. In the tail-suspended animals, a more complete repair was observed with nonenlarged area of nonmuscle cells or matrix material visible. In contrast, the muscle samples from the flight animals were less well organized and contained more macrophages and blood vessels in the repair region, indicative of a delayed repair process, but did not demonstrate any chronic inflammation. Myofiber repair did vary in muscles from the different groups, being slowest in the flight animals and most complete in the tail-suspended ones.

Stauber, W. T.↗