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At least 19 records

Comparison of stress tolerance mechanisms between Saccharomyces cerevisiae and the multistress-tolerant Pichia kudriavzevii

Yeasts play a vital role in both research and industrial biomanufacturing. Saccharomyces cerevisiae has been extensively utilized as a model system. However, its application is often constrained by limited tolerance to the diverse stress conditions encountered in bioprocesses. These challenges have driven increasing interest in nonconventional, multistress-tolerant yeasts as alternative biomanufacturing hosts. This review highlights Pichia kudriavzevii as a promising nonconventional yeast for industrial applications. Unlike S. cerevisiae, P. kudriavzevii exhibits exceptional tolerance to high temperatures, elevated concentrations of furanic and phenolic inhibitors, osmotic stress, salinity, and extreme pH. These traits make it an attractive candidate for industrial processes without requiring extensive genetic modifications to enhance stress resistance. As a result, P. kudriavzevii has emerged as a flagship species for advancing bioeconomy. Despite its industrial potential, the molecular mechanisms underlying P. kudriavzevii's superior stress tolerance remain poorly understood. This review compiles current knowledge on P. kudriavzevii and compares its stress tolerance mechanisms with those of S. cerevisiae, providing insights into its innate resilience. By expanding our understanding of nonconventional yeasts, this review aims to facilitate their broader adoption as robust microbial platforms for industrial biomanufacturing.

Frousnoon, Thasneem Banu

Contrasting effects of glutamate and branched-chain amino acid metabolism on acid tolerance in a Castellaniella isolate from acidic groundwater

Groundwater acidification co-occurring with nitrate pollution is a common, global environmental health hazard. Denitrifying bacteria have been leveraged for the in situ removal of nitrate in groundwater. However, co-existing stressors—such as low pH—reduce the efficacy of biological removal processes. Castellaniella sp. str. MT123 is a complete denitrifier that was isolated from acidic, nitrate-contaminated groundwater. The strain grows robustly by nitrate respiration at pH < 6.0, completely reducing nitrate to dinitrogen gas. Genomic analyses of MT123 revealed few previously characterized acid tolerance genes. Thus, we utilized a combination of proteomics, metabolomics, and competitive mutant fitness to characterize the genetic mechanisms of MT123 acclimation to growth under mildly acidic conditions. We found that glutamate accumulation is critical in the acid acclimation of MT123, possibly through consumption of intracellular protons via glutamate decarboxylation to GABA. This is despite the fact that MT123 lacks the canonical glutamate decarboxylase-glutamate/GABA antiporter system implicated in acid tolerance in other bacteria. In contrast, branched-chain amino acid (BCAA) accumulation was detrimental to cell growth at lower pHs, possibly through indirect mechanisms impacting the cellular glutamate pool. Genetic analysis previously linked MT123 to a population of Castellaniella that bloomed—concurrent to nitrate removal—during a biostimulation effort to reduce groundwater nitrate concentrations at MT123’s location of origin. Thus, our analyses provide novel insight into mechanisms of acclimation to acidic conditions in a strain with significant potential for nitrate bioremediation.

59 BASIC BIOLOGICAL SCIENCES

Data for "Genetics of flooding tolerance in an F2 Miscanthus sacchariflorus ssp. lutarioriparius × M. sinensis population"

This dataset contains all data and supplementary materials from "Genetics of flooding tolerance in an F2 Miscanthus sacchariflorus ssp. lutarioriparius × M. sinensis population". 1. The dataset S1 table contains the raw phenotypic data collected during the experiment. 2. The dataset S2 table contains the LSmean values for the 24 traits studied. 3. The dataset S3 table contains the TASSEL GBSv2 map, marker information, and genotype data used for mapping. 4. The dataset S4 table contains information on candidate genes found in each of the QTL intervals. 5. The dataset S5 table contains the GO annotations and KEGG enrichment analyses for those candidate genes. 6. The dataset S6 table contains information on the sequences used to classify AP2 ERF transcription factors. 7. The dataset S7 table contains information on AP2 ERF orthologs between Miscanthus and rice based on synteny. 8. Supplementary file 1 contains the ANOVA results using the raw phenotypic data collected from protocol "A". 9. Supplementary file 2 contains the ANOVA results using the raw phenotypic data collected from protocol "B". 10. Supplementary file 3 contains notes on the comparison of SNP calling methods. 11. Supplementary file 4 is a script for analyzing candidate genes found in QTL intervals.

Miscanthus, flood, partial submergence, complete s

The software-implemented fault tolerance /SIFT/ approach to fault tolerant computing

SIFT is an experimental computer designed for highly reliable flight-control service in advanced air transports. Its development was intended to integrate and demonstrate the latest techniques in fault-tolerant computing. During its development, several new problems of some generality were uncovered and solved. The technology developed for the validation of its design is seen as being perhaps as important as the design itself. The SIFT design is described, as is the way in which the design and its validation were shaped by the requirements of its intended application. Attention is also given to reliability and fault tolerance. The most significant feature of the hardware design is the absence of elements that can generate multiple faults, such as shared clocks or data buses. It is noted that the software is realized in only 800 lines of code, of which 80% are in a high-level language.

Goldberg, J.

Rapid recovery from transient faults in the fault-tolerant processor with fault-tolerant shared memory

The Draper fault-tolerant processor with fault-tolerant shared memory (FTP/FTSM), which is designed to allow application tasks to continue execution during the memory alignment process, is described. Processor performance is not affected by memory alignment. In addition, the FTP/FTSM incorporates a hardware scrubber device to perform the memory alignment quickly during unused memory access cycles. The FTP/FTSM architecture is described, followed by an estimate of the time required for channel reintegration.

Harper, Richard E.

Advanced information processing system: The Army fault tolerant architecture conceptual study. Volume 2: Army fault tolerant architecture design and analysis

Described here is the Army Fault Tolerant Architecture (AFTA) hardware architecture and components and the operating system. The architectural and operational theory of the AFTA Fault Tolerant Data Bus is discussed. The test and maintenance strategy developed for use in fielded AFTA installations is presented. An approach to be used in reducing the probability of AFTA failure due to common mode faults is described. Analytical models for AFTA performance, reliability, availability, life cycle cost, weight, power, and volume are developed. An approach is presented for using VHSIC Hardware Description Language (VHDL) to describe and design AFTA's developmental hardware. A plan is described for verifying and validating key AFTA concepts during the Dem/Val phase. Analytical models and partial mission requirements are used to generate AFTA configurations for the TF/TA/NOE and Ground Vehicle missions.

Harper, R. E.

Advanced information processing system: The Army fault tolerant architecture conceptual study. Volume 1: Army fault tolerant architecture overview

Digital computing systems needed for Army programs such as the Computer-Aided Low Altitude Helicopter Flight Program and the Armored Systems Modernization (ASM) vehicles may be characterized by high computational throughput and input/output bandwidth, hard real-time response, high reliability and availability, and maintainability, testability, and producibility requirements. In addition, such a system should be affordable to produce, procure, maintain, and upgrade. To address these needs, the Army Fault Tolerant Architecture (AFTA) is being designed and constructed under a three-year program comprised of a conceptual study, detailed design and fabrication, and demonstration and validation phases. Described here are the results of the conceptual study phase of the AFTA development. Given here is an introduction to the AFTA program, its objectives, and key elements of its technical approach. A format is designed for representing mission requirements in a manner suitable for first order AFTA sizing and analysis, followed by a discussion of the current state of mission requirements acquisition for the targeted Army missions. An overview is given of AFTA's architectural theory of operation.

Harper, R. E.

Tolerance and UQ4SIM: Nimble Uncertainty Documentation and Analysis Software

Ultimately, scientific numerical models need quantified output uncertainties so that modeling can evolve to better match reality. Documenting model input uncertainties and variabilities is a necessary first step toward that goal. Without known input parameter uncertainties, model sensitivities are all one can determine, and without code verification, output uncertainties are simply not reliable. The basic premise of uncertainty markup is to craft a tolerance and tagging mini-language that offers a natural, unobtrusive presentation and does not depend on parsing each type of input file format. Each file is marked up with tolerances and optionally, associated tags that serve to label the parameters and their uncertainties. The evolution of such a language, often called a Domain Specific Language or DSL, is given in [1], but in final form it parallels tolerances specified on an engineering drawing, e.g., 1 +/- 0.5, 5 +/- 10%, 2 +/- 10 where % signifies percent and o signifies order of magnitude. Tags, necessary for error propagation, can be added by placing a quotation-mark-delimited tag after the tolerance, e.g., 0.7 +/- 20% 'T_effective'. In addition, tolerances might have different underlying distributions, e.g., Uniform, Normal, or Triangular, or the tolerances may merely be intervals due to lack of knowledge (uncertainty). Finally, to address pragmatic considerations such as older models that require specific number-field formats, C-style format specifiers can be appended to the tolerance like so, 1.35 +/- 10U_3.2f. As an example of use, consider figure 1, where a chemical reaction input file is has been marked up to include tolerances and tags per table 1. Not only does the technique provide a natural method of specifying tolerances, but it also servers as in situ documentation of model uncertainties. This tolerance language comes with a utility to strip the tolerances (and tags), to provide a path to the nominal model parameter file. And, as shown in [1], having the ability to quickly mark and identify model parameter uncertainties facilitates error propagation, which in turn yield output uncertainties.

Kleb, Bil

A small number of point mutations confer formate tolerance in Shewanella oneidensis

ABSTRACT Microbial electrosynthesis (MES) is a sustainable approach to chemical production from CO 2 and clean electricity. However, limitations in electron transfer efficiency and gaps in understanding of electron transfer pathways in MES systems prevent full realization of this technology. Shewanella oneidensis could serve as an MES biocatalyst because it has a well-studied, efficient transmembrane electron transfer pathway. A key first step in MES in this organism could be CO 2 reduction to formate. However, we report that wild-type S. oneidensis does not tolerate high levels of formate. In this work, we created and characterized formate-tolerant strains of S. oneidensis for further engineering and future use in MES systems through adaptive laboratory evolution. Two different point mutations in a gene encoding a predicted sodium-dependent bicarbonate transporter and a DUF2721-containing protein separately confer formate tolerance to S. oneidensis . The mutations were further evaluated to understand their role in improving formate tolerance. We also show that the wild-type and mutant versions of the putative sodium-dependent bicarbonate transporter improve formate tolerance of Zymomonas mobilis , indicating the potential of transferring this formate tolerance phenotype to other organisms. IMPORTANCE Shewanella oneidensis is a bacterium with a well-studied, efficient extracellular electron transfer pathway. This capability could make this organism a suitable host for microbial electrosynthesis using CO 2 or formate as feedstocks. However, we report here that formate is toxic to S. oneidensis , limiting the potential for its use in these systems. In this work, we evolve several strains of S. oneidensis that have improved formate tolerance, and we investigate some mutations that confer this phenotype. The phenotype is confirmed to be attributed to several single point mutations by transferring the wild-type and mutant versions of each gene to the wild-type strain. Finally, the formate tolerance mechanism of one variant is studied using structural modeling and expression in another host. This study, therefore, presents a simple method for conferring formate tolerance to bacterial hosts.

Cross, Megan C. Gruenberg (ORCID:0000000291589900)

Alternate routes to acetate tolerance lead to varied isoprenol production from mixed carbon sources in Pseudomonas putida

ABSTRACT Lignocellulose is a renewable resource for the production of a diverse array of platform chemicals, including the biofuel isoprenol. Although this carbon stream provides a rich source of sugars, other organic compounds, such as acetate, can be used by microbial hosts. Here, we examined the growth and isoprenol production in a Pseudomonas putida strain pre-tolerized (“PT”) background where its native isoprenol catabolism pathway is deleted, using glucose and acetate as carbon sources. We found that PT displays impaired growth in minimal medium containing acetate and often fails to grow in glucose-acetate medium. Using a mutant recovery-based approach, we generated tolerized strains that overcame these limitations, achieving fast growth and isoprenol production in the mixed carbon feed. Changes in the glucose and acetate assimilation routes, including an upregulation in PP_0154 (SpcC, succinyl-CoA:acetate CoA-transferase) and differential expression of the gluconate assimilation pathways, were key for higher isoprenol titers in the tolerized strains, whereas a different set of mechanisms were likely enabling tolerance phenotypes in media containing acetate. Among these, a coproporphyrinogen-III oxidase (HemN) was upregulated across all tolerized strains and in one isolate required for acetate tolerance. Utilizing a defined glucose and acetate mixture ratio reflective of lignocellulosic feedstocks for isoprenol production in P. putida allowed us to obtain insights into the dynamics and challenges unique to dual carbon source utilization that are obscured when studied separately. Together, this enabled the development of a P. putida bioconversion chassis able to use a more complex carbon stream to produce isoprenol. IMPORTANCE Acetate is a relatively abundant component of many lignocellulosic carbon streams and has the potential to be used together with sugars, especially in microbes with versatile catabolism such as P. putida . However, the use of mixed carbon streams necessitates additional optimization. Furthermore, the use of P. putida for the production of the biofuel target, isoprenol, requires the use of engineered strains that have additional growth and production constraints when cultivated in acetate and glucose mixtures. In this study, we generate acetate-tolerant P. putida strains that overcome these challenges and examine their ability to produce isoprenol. We show that acetate tolerance and isoprenol production, although independent phenotypes, can both be optimized in a given P. putida strain. Using proteomics and whole genome sequencing, we examine the molecular basis of both phenotypes and show that tolerance to acetate can occur via alternate routes and result in different impacts on isoprenol production.

de Siqueira, Guilherme M. V. (ORCID:00000002364563

Effects of cholinergic and beta-adrenergic blockade on orthostatic tolerance in healthy subjects

Cardiovascular responses during a graded lower body negative pressure (LBNP) protocol were compared before and after atropine and propranolol administration to test the hypothesis that both sympathetic and parasympathetic control of cardio-acceleration are associated with syncopal predisposition to orthostatic stress in healthy subjects. Eleven men were categorized into two groups having high (HT, N = 6) or low (LT, N = 5) tolerance based on their total time before the onset of presyncopal symptoms. HT and LT groups were similar in physical characteristics, fitness, and baseline cardiovascular measurements. Atropine treatment had no effect on LBNP tolerance or mean arterial pressure at presyncope, despite an atropine-induced increase in heart rate. Propranolol treatment reduced (p<0.05) LBNP tolerance in both groups. Diminished LBNP tolerance after propranolol administration was associated with reductions in cardiac output, whereas increase in systemic peripheral resistance from baseline to presyncope was unaffected by propranolol. Reduction in cardiac output and LBNP tolerance after beta blockade reflected a chronotropic effect because lower LBNP tolerance for the HT (-50%) and LT (-39%) groups was associated with dramatic reductions (p <0.05) in the magnitude of LBNP-induced tachycardia without significant effects on stroke volume at presyncope. Absence of an atropine-induced difference in cardiac output and systemic peripheral resistance between HT and LT groups failed to support the notion that cardiac vagal withdrawal represents a predominant mechanism that could account for differences in orthostatic tolerance. Because a reduction in LBNP tolerance in both HT and LT groups after propranolol treatment was most closely associated with reduced tachycardia, the data suggest that a primary autonomically mediated mechanism for maintenance of mean arterial pressure and orthostatic tolerance in healthy subjects is beta adrenergic-induced tachycardia.

Non-NASA Center

Issues on human acceleration tolerance after long-duration space flights

This report reviewed the literature on human tolerance to acceleration at 1 G and changes in tolerance after exposure to hypogravic fields. It was found that human tolerance decreased after exposure to hypokinetic and hypogravic fields, but the magnitude of such reduction ranged from 0 to 30 percent for plateau G forces and 30 to 70 percent for time tolerance on sustained G forces. A logistic regression model of the probability of individuals with 25 percent reduction in +Gz tolerance after 1 to 41 days of hypogravic exposures was constructed. The estimated values from the model showed a good correlation with the observed data. A brief review of the need for in-flight centrifuge during long-duration missions was also presented. Review of the available data showed that the use of countermeasures (such as anti-G suits, periodic acceleration, and exercise) reduced the decrement in acceleration tolerance after long-duration space flights. Areas of further research include quantification of the effect of countermeasures on tolerance, and methods to augment tolerance during and after exposures to hypogravic fields. Such data are essential for planning long-duration human missions.

Kumar, K. Vasantha

On the design of fault-tolerant robotic manipulator systems

Robotic systems are finding increasing use in space applications. Many of these devices are going to be operational on board the Space Station Freedom. Fault tolerance has been deemed necessary because of the criticality of the tasks and the inaccessibility of the systems to maintenance and repair. Design for fault tolerance in manipulator systems is an area within robotics that is without precedence in the literature. In this paper, we will attempt to lay down the foundations for such a technology. Design for fault tolerance demands new and special approaches to design, often at considerable variance from established design practices. These design aspects, together with reliability evaluation and modeling tools, are presented. Mechanical architectures that employ protective redundancies at many levels and have a modular architecture are then studied in detail. Once a mechanical architecture for fault tolerance has been derived, the chronological stages of operational fault tolerance are investigated. Failure detection, isolation, and estimation methods are surveyed, and such methods for robot sensors and actuators are derived. Failure recovery methods are also presented for each of the protective layers of redundancy. Failure recovery tactics often span all of the layers of a control hierarchy. Thus, a unified framework for decision-making and control, which orchestrates both the nominal redundancy management tasks and the failure management tasks, has been derived. The well-developed field of fault-tolerant computers is studied next, and some design principles relevant to the design of fault-tolerant robot controllers are abstracted. Conclusions are drawn, and a road map for the design of fault-tolerant manipulator systems is laid out with recommendations for a 10 DOF arm with dual actuators at each joint.

Tesar, Delbert

CORSSTOL: Cylinder Optimization of Rings, Skin, and Stringers with Tolerance sensitivity

Cylinder Optimization of Rings, Skin, and Stringers with Tolerance (CORSSTOL) sensitivity is a design optimization program incorporating a method to examine the effects of user-provided manufacturing tolerances on weight and failure. CORSSTOL gives designers a tool to determine tolerances based on need. This is a decisive way to choose the best design among several manufacturing methods with differing capabilities and costs. CORSSTOL initially optimizes a stringer-stiffened cylinder for weight without tolerances. The skin and stringer geometry are varied, subject to stress and buckling constraints. Then the same analysis and optimization routines are used to minimize the maximum material condition weight subject to the least favorable combination of tolerances. The adjusted optimum dimensions are provided with the weight and constraint sensitivities of each design variable. The designer can immediately identify critical tolerances. The safety of parts made out of tolerance can also be determined. During design and development of weight-critical systems, design/analysis tools that provide product-oriented results are of vital significance. The development of this program and methodology provides designers with an effective cost- and weight-saving design tool. The tolerance sensitivity method can be applied to any system defined by a set of deterministic equations.

Finckenor, J.

The Evolution of Sulfide Tolerance in the Cyanobacteria

Understanding how the function of extant microorganisms has recorded both their evolutionary histories and their past interactions with the environment is a stated goal of astrobiology. We are taking a multidisciplinary approach to investigate the diversification of sulfide tolerance mechanisms in the cyanobacteria, which vary both in their degree of exposure to sulfide and in their capacity to tolerate this inhibitor of photosynthetic electron transport. Since conditions were very reducing during the first part of Earth's history and detrital sulfides have been found in Archean sediments, mechanisms conferring sulfide tolerance may have been important for the evolutionary success of the ancestors of extant cyanobacteria. Two tolerance mechanisms have been identified in this group: (1) resistance of photosystem II, the principal target of sulfide toxicity; and (2) maintenance of the ability to fix carbon despite photosystem II inhibition by utilizing sulfide as an electron donor in photosystem I - dependent, anoxygenic photosynthesis. We are presently collecting comparative data on aspects of sulfide physiology for laboratory clones isolated from a variety of habitats. These data will be analyzed within a phylogenetic framework inferred from molecular sequence data collected for these clones to test how frequently different mechanisms of tolerance have evolved and which tolerance mechanism evolved first. In addition, by analyzing these physiological data together with environmental sulfide data collected from our research sites using microelectrodes, we can also test whether the breadth of an organism's sulfide tolerance can be predicted from the magnitude of variation in environmental sulfide concentration it has experienced in its recent evolutionary past and whether greater average sulfide concentration and/or temporal variability in sulfide favors the evolution of a particular mechanism of sulfide tolerance.

Miller, Scott R.

Eigenstructure Assignment for Fault Tolerant Flight Control Design

In recent years, fault tolerant flight control systems have gained an increased interest for high performance military aircraft as well as civil aircraft. Fault tolerant control systems can be described as either active or passive. An active fault tolerant control system has to either reconfigure or adapt the controller in response to a failure. One approach is to reconfigure the controller based upon detection and identification of the failure. Another approach is to use direct adaptive control to adjust the controller without explicitly identifying the failure. In contrast, a passive fault tolerant control system uses a fixed controller which achieves acceptable performance for a presumed set of failures. We have obtained a passive fault tolerant flight control law for the F/A-18 aircraft which achieves acceptable handling qualities for a class of control surface failures. The class of failures includes the symmetric failure of any one control surface being stuck at its trim value. A comparison was made of an eigenstructure assignment gain designed for the unfailed aircraft with a fault tolerant multiobjective optimization gain. We have shown that time responses for the unfailed aircraft using the eigenstructure assignment gain and the fault tolerant gain are identical. Furthermore, the fault tolerant gain achieves MIL-F-8785C specifications for all failure conditions.

Sobel, Kenneth

Low-overhead transversal fault tolerance for universal quantum computation

Fast, reliable logical operations are essential for realizing useful quantum computers. By redundantly encoding logical qubits into many physical qubits and using syndrome measurements to detect and correct errors, we can achieve low logical error rates. However, for many practical quantum error correction codes such as the surface code, owing to syndrome measurement errors, standard constructions require multiple extraction rounds—of the order of the code distance d—for fault-tolerant computation, particularly considering fault-tolerant state preparation. Here we show that logical operations can be performed fault-tolerantly with only a constant number of extraction rounds for a broad class of quantum error correction codes, including the surface code with magic state inputs and feedforward, to achieve ‘transversal algorithmic fault tolerance’. Through the combination of transversal operations7 and new strategies for correlated decoding, despite only having access to partial syndrome information, we prove that the deviation from the ideal logical measurement distribution can be made exponentially small in the distance, even if the instantaneous quantum state cannot be made close to a logical codeword because of measurement errors. We supplement this proof with circuit-level simulations in a range of relevant settings, demonstrating the fault tolerance and competitive performance of our approach. Furthermore, our work sheds new light on the theory of quantum fault tolerance and has the potential to reduce the space–time cost of practical fault-tolerant quantum computation by over an order of magnitude.

Zhou, Hengyun [QuEra Computing, Boston, MA (United

Impact of mechanical tolerances on partial turn skipping in helical flux compression generator

Maintaining tight mechanical tolerances of the components used in helical flux compression generators is crucial for optimizing device performance. Moving beyond current literature, which provides various approximations for acceptable tolerances, a 3D simulation of the armature stator interaction during generator operation was developed, revealing that existing equations frequently misestimate the tolerance limits. Without restricting the generality of the presented approach, the focus was primarily on the armature’s concentricity and roundness, comparing the acceptable tolerance limits between generators of different sizes. An analytical approach for select geometries was developed, confirming the 3D results. Quantitative results reveal that a steeper Gurney angle and increased winding pitch allow for less restrictive tolerances for eccentricity and ellipticity. Even small deviations from the ideal armature positioning and shape will result in fluctuations of the output current time-derivative, observed experimentally and in simulation. Larger deviations then cause partial turn skipping, associated with loss of magnetic flux. The simulation elucidates the impact of tolerance deviations on generator performance by offering precise tolerance ranges for various generator sizes, thereby facilitating informed design decisions.

42 ENGINEERING