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Metabolite sensing is a fundamental biological process, and the perception of dynamic changes in the extracellular environment is of paramount importance for the survival of organisms. Bacteria usually adjust their metabolisms to changing environments via transcriptional regulation.
The galactose/glucose-binding protein (GBP) is synthesized in the cytoplasm of Escherichia coli in a precursor form and exported into the periplasmic space upon cleavage of a 23-amino-acid leader sequence. GBP binds galactose and glucose in a highly specific manner. The ligand induces a hinge motion in GBP and the resultant protein conformational change constitutes the basis of the sensing system. The mglB gene, which codes for GBP, was isolated from the chromosome of E. coli using the polymerase chain reaction (PCR). Since wild-type GBP lacks cysteines in its structure, introducing this amino acid by site-directed mutagenesis ensures single-label attachment at specific sites with a sulfhydro-specific fluorescent probe. Site-directed mutagenesis by overlap extension PCR was performed to prepare three different mutants to introduce a single cysteine residue at positions 148, 152, and 182. Since these residues are not involved in ligand binding and since they are located at the edge of the binding cleft, they experience a significant change in environment upon binding of galactose or glucose. The sensing system strategy is based on the fluorescence changes of the probe as the protein undergoes a structural change on binding. In this work a reagentless sensing system has been rationally designed that can detect submicromolar concentrations of glucose. The calibration plots have a linear working range of three orders of magnitude. Although the system can sense galactose as well, this epimer is not a potential interfering substance since its concentration in blood is negligible. Copyright 2001 Academic Press.
As an integral part of the National Climate Assessment (NCA), technical assessment reports for 13 regions in the U.S. that describe the scientific rationale to support climate change impacts within the purview of these regions, and provide adaptation or mitigation measures in response to these impacts. These technical assessments focus on climate change impacts on sectors that are important environmental, biophysical, and social and economic aspects of sustainability within the U.S.: Climate change science, Ecosystems and biodiversity, Water resources, Human health, Energy supply and use, Water/energy/land use, Transportation, Urban/infrastructure/vulnerability, Agriculture, Impacts of climate change on tribal/indigenous and native lands and resources, Forestry, Land use/land cover change, Rural communities development, and Impacts on biogeochemical cycles, with implications for ecosystems and biodiversity. There is a critical and timely need for the development of mitigation and adaptation strategies in response to climate change by the policy and decision making communities, to insure resiliency and sustainability of the built environment in the future.
Over the last 15 years a striking pattern of diversification has been documented in the fossil record of benthic marine invertebrates. Higher taxa (orders) tend to originate onshore, diversify offshore, and retreat into deep-water environments. Previous studies attribute this macroevolutionary pattern to a variety of causes, foremost among them the role of nearshore disturbance in providing opportunities for the evolution of novel forms accorded ordinal rank. Our analysis of the post-Paleozoic record of ordinal first appearances indicates that the onshore preference of ordinal origination occurred only in the Mesozoic prior to the Turonian stage of the Cretaceous, a period characterized by relatively frequent anoxic/dysoxic bottom conditions in deeper marine environments. Later, in the Cretaceous and Cenozoic, ordinal origination of benthic organisms did not occur exclusively, or even preferentially, in onshore environments. This change in environmental pattern of ordinal origination roughly correlates with Late Cretaceous: (i) decline in anoxia/dysoxia in offshore benthic environments; (ii) extinction of faunas associated with dysoxic conditions; (iii) increase in bioturbation with the expansion of deep burrowing forms into offshore environments; and (iv) offshore expansion of bryozoan diversity. We also advance a separate argument that the Cenomanian/Turonian and latest Paleocene global events eliminated much of the deep-water benthos. This requires a more recent origin of modern vent and deep-sea faunas, from shallower water refugia, than the Paleozoic or early Mesozoic origin of these faunas suggested by other workers.
Many microbial pathogen shave repeatedly exhibited unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, host-pathogen interactions, and virulence. Notably, the classic foodborne pathogen Salmonella enterica serovar Typhimurium displayed increased virulence in animals when cultured in either the spaceflight analogue or true spaceflight environment. Recently, Serratia marcescens also was shown to increase virulence when cultured in the spaceflight environment. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system at multiple levels, which suggests an increased risk of infectious diseases. Moreover, astronauts have some degree of clinical infectious disease incidence. However, the contribution of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. The goal of this study is to gain insight into the breadth of other medically significant microbial pathogens that may exhibit altered virulence and pathogenesis-related responses when cultured in space flight analogue conditions. Specifically, we are characterizing the effect of spaceflight analogue culture (Low Shear Modeled Microgravity/LSMMG) on microbial pathogenesis-related stress responses, in vitro host-pathogen interactions, gene expression, and virulence potential in animals using five important model bacterial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. Herein, we present data from one of these pathogens, the foodborne bacterium, S. enterica Enteritidis, which is closely related to S. enterica Typhimurium. Phenotypes evaluated included growth profiles, environmental stress responses(acid, oxidative, bile, and thermal stresses),and in vitro colonization of3-D biomimetic cultures of human intestinal tissue containing immune cells. Transcriptomic profiling and virulence studies are ongoing. We show that S. Enteritidis exhibited key alterations in pathogenic responses to LSMMG culture that suggest increased infection risk, including several responses which were different from those observed in the closely related pathovar S. Typhimurium. This information will provide critical mechanistic insight into the potential impact of microgravity on alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.
Many microbial pathogen shave repeatedly exhibited unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, host-pathogen interactions, and virulence. Notably, the classic foodborne pathogen Salmonella enterica serovar Typhimurium displayed increased virulence in animals when cultured in either the spaceflight analogue or true spaceflight environment. Recently, Serratia marcescens also was shown to increase virulence when cultured in the spaceflight environment. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system at multiple levels, which suggests an increased risk of infectious diseases. Moreover, astronauts have some degree of clinical infectious disease incidence. However, the contribution of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. The goal of this study is to gain insight into the breadth of other medically significant microbial pathogens that may exhibit altered virulence and pathogenesis-related responses when cultured in space flight analogue conditions. Specifically, we are characterizing the effect of spaceflight analogue culture (Low Shear Modeled Microgravity/LSMMG) on microbial pathogenesis-related stress responses, in vitro host-pathogen interactions, gene expression, and virulence potential in animals using five important model bacterial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. Herein, we present data from one of these pathogens, the foodborne bacterium, S. enterica Enteritidis, which is closely related to S. enterica Typhimurium. Phenotypes evaluated included growth profiles, environmental stress responses(acid, oxidative, bile, and thermal stresses),and in vitro colonization of3-D biomimetic cultures of human intestinal tissue containing immune cells. Transcriptomic profiling and virulence studies are ongoing. We show that S. Enteritidis exhibited key alterations in pathogenic responses to LSMMG culture that suggest increased infection risk, including several responses which were different from those observed in the closely related pathovar S. Typhimurium. This information will provide critical mechanistic insight into the potential impact of microgravity on alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.
Many microbial pathogens have repeatedly exhibited unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, host-pathogen interactions, and virulence. Notably, the classic foodborne pathogen Salmonella enterica serovar Typhimurium displayed increased virulence in animals when cultured in either the spaceflight analogue or true spaceflight environment. Recently, Serratia marcescens also was shown to increase virulence when cultured in the spaceflight environment. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system at multiple levels, which suggests an increased risk of infectious diseases. Moreover, astronauts have some degree of clinical infectious disease incidence. However, the contribution of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. The goal of this study is to gain insight into the breadth of other medically significant microbial pathogens that may exhibit altered virulence and pathogenesis-related responses when cultured in spaceflight analogue conditions. Specifically, we are characterizing the effect of spaceflight analogue culture (Low Shear Modeled Microgravity/LSMMG) on microbial pathogenesis-related stress responses, in vitro host-pathogen interactions, gene expression, and virulence potential in animals using five important model bacterial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. The information to date is providing a better understanding into the potential impact of microgravity on alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.
Autonomous systems are complex, diverse, and likely be used in environments that change unpredictably. We must develop mechanisms to cope with failures, potentially caused by both the natural stresses of the operating environment and the system's inadequate environmental models. Failures also arise from system component wear. In 2016, the IEEE launched the Global Initiative on Ethics of Autonomous and Intelligent Systems in response to the proliferation of increasingly-autonomous systems. A cornerstone of this initiative is the IEEE P7000 series of projects, which set the standards for the future of ethical intelligent and autonomous technologies. This paper describes the ongoing development of the IEEE P7009 standard on “Fail-Safe Design of Autonomous and Semi-Autonomous Systems”. We provide a collective perspective on the ongoing development of IEEE P7009, and outline the objectives, current approaches, issues under discussion, and two exemplar use cases from different sectors: one high-regulation, the other low-regulation.
INTRODUCTION Over the past 50 years, microorganisms have displayed unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in final cell concentration, biofilm production, stress resistance, antibiotic sensitivity, gene expression, and virulence. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system; consisting of altered leukocyte distribution, reductions in T and NK cell function, altered cytokine profiles, and reactivation of latent herpesviruses. Further, astronauts have some degree of clinical incidence, primarily infectious disease episodes and atopic dermatitis. The impact of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. SPECIFIC AIMS In the Host-Microorganism, Microbial Response aspect of this study, the following Specific Aims are being investigated, using the microbial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli. Aim 1: Characterize the effect of spaceflight analogue culture on microbial pathogenesis-related stress responses and in vitro host-pathogen interactions. Analysis will include microbial stress responses as well as colonization and viability following pathogen challenge of three-dimensional (3-D) tissue co-culture models containing immune cells. We specifically selected obligate and opportunistic pathogens that are medically important and have been or are likely to be found aboard spacecraft. Aim 2: Characterize the effect of spaceflight analogue culture on the virulence potential of pathogenic microorganisms. Changes in virulence will be assessed using a mouse model of infection. SIGNIFICANCE The goal of this study is to use spaceflight analogue conditions to gain insight into the breadth of medically-significant International Space Station microorganisms that have altered virulence and the impact of those changes on the immune response of the host, including astronaut immune cells. This information will provide critical understanding into the impact of microgravity on potential alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.
KSC has developed an environmental program to ensure that its activities do not adversely affect the surrounding environment. Two essential elements of the total program are the baseline and monitoring programs. The goal of the baseline program is to collect sufficient information about the environment prior to Shuttle launches so that adverse changes in the environment - if and when they occur after the Shuttle program becomes active - can be detected and cause-effect relationships established when possible. The goal of the monitoring program is to use information from the baseline program along with survey and sampling operations during the period of initial Shuttle launches to document adverse changes in the environment.
INTRODUCTION Over the past 50 years, many microorganisms have displayed unexpected responses relevant to infectious disease when grown in microgravity and microgravity analogue environments, including changes in stress resistance, biofilm production, antibiotic sensitivity, final cell concentration, gene expression, enhanced host-pathogen interaction, and virulence. In parallel, astronaut studies have characterized a persistent spaceflight-induced dysregulation of the human immune system; consisting of altered leukocyte distribution, reductions in T and NK cell function, altered cytokine profiles, and reactivation of latent herpesviruses. Further, astronauts have some degree of clinical incidence, primarily infectious disease episodes and atopic dermatitis. The impact of the microgravity environment on host-pathogen interactions and potential for clinical disease remains understudied and poorly characterized. SPECIFIC AIMS In this study, the following Specific Aims are being investigated, using the microbial pathogens, Salmonella enterica Enteritidis, Pseudomonas aeruginosa, Burkholderia cepacia, Streptococcus pneumoniae, and enterohemorrhagic Escherichia coli (EHEC). Aim 1: Characterize the effect of spaceflight analogue culture on microbial pathogenesis-related stress responses and in vitro host-pathogen interactions. Analyses include microbial stress responses as well as colonization and viability following pathogen challenge of three-dimensional (3-D) tissue co-culture models containing immune cells. We specifically selected obligate and opportunistic pathogens that are medically important and have been or are likely to be found aboard spacecraft. Aim 2: Characterize the effect of spaceflight analogue culture on the virulence potential of pathogenic microorganisms. Virulence will be assessed using a mouse model of infection. SIGNIFICANCE The goal of this study is to use spaceflight analogue conditions to gain insight into the breadth of medically-significant International Space Station microorganisms that have altered virulence and the impact of those changes on the immune response of the host. This information will provide critical understanding into the impact of microgravity on potential alterations in microbial virulence and associated infectious disease risk to crew health during spaceflight missions.
The Mars Science Laboratory (MSL) Curiosity rover has spent the last two years investigating a prominent resistant ridge, informally named the Vera Rubin Ridge (VRR), at the base of Mount Sharp (Aeolis Mons). The ridge has been a high priority science target for the MSL mission since landing in Gale crater more than 6 years ago because of the detection of a strong hematite spectral signature, and its distinct topography. Examining the chemistry of the ridge can aid in determining the relationship to other rocks analyzed during the rover traverse, specifically the Murray formation (fm) encountered below the ridge. We can also determine compositional trends with elevation and/or laterally within the ridge, and whether spectral properties observed on the ridge, both from orbit and in situ, correspond with changes in chemistry. The composition of the ridge, combined with mineralogy of drilled samples, can help to elucidate bigger picture questions regarding depositional environment, possible changing lake water chemistry and diagenetic/alteration history.
The International Space Station (ISS) program utilizes a plasma environment forecast for estimating electrical charging hazards for crews during extravehicular activity (EVA). The process uses ionospheric electron density (Ne) and temperature (Te) measurements from the ISS Floating Potential Measurement Unit (FPMU) instrument suite with the assumption that the plasma conditions will remain constant for one to fourteen days with a low probability for a space weather event which would significantly change the environment before an EVA. FPMU data is typically not available during EVA's, therefore, the most recent FPMU data available for characterizing the state of the ionosphere during EVA is typically a day or two before the start of an EVA or after the EVA has been completed. Three near real time space weather tools under development for ISS applications are described here including: (a) Ne from ground based ionosonde measurements of foF2 (b) Ne from near real time satellite radio occultation measurements of electron density profiles (c) Ne, Te from a physics based ionosphere model These applications are used to characterize the ISS space plasma environment during EVA periods when FPMU data is not available, monitor for large changes in ionosphere density that could render the ionosphere forecast and plasma hazard assessment invalid, and validate the "persistence of conditions" forecast assumption. In addition, the tools are useful for providing space environment input to science payloads on ISS and anomaly investigations during periods the FPMU is not operating.
The built environment consists of components that exist at a range of scales from small (e.g., houses, shopping malls) to large (e.g., transportation networks) to highly modified landscapes such as cities. Thus, the impacts of climate change on the built environment may have a multitude of effects on humans and the land. The impact of climate change may be exacerbated by the interaction of different events that singly may be minor, but together may have a synergistic set of impacts that are significant. Also, mechanisms may exist wherein the built environment, particularly in the form of cities, may affect weather and the climate on local and regional scales. Hence, a city may be able to cope with prolonged heat waves, but if this is combined with severe drought, the overall result could be significant or even catastrophic, as accelerating demand for energy to cooling taxes water supplies needed both for energy supply and municipal water needs. This presentation surveys potential climate change impacts on the built environment from the perspective of the National Climate Assessment, and explores adaptation measures that can be employed to mitigate these impacts.
The theory of evolution through natural selection was proposed by Darwin and Wallace to explain how the characteristics of populations of animals change with time. An examination of their assumptions shows that the theory has much broader application than they originally envisaged. We now know that in appropriate environments RNA molecules or computer viruses, for example, can evolve. The adventure with which we are concerned is the quest for chemical systems that undergo processes analogous to Darwinian selection in the test tube. The search is not restricted to systems that are closely related to nucleic acids, although most of the available experimental evidence concerns such systems. A population of molecules satisfies all the requirements of the theory is there are different kinds of molecules in the population and if each individual molecule can direct the formation of copies of itself, then a population of molecules will adapt to a varying environment by changing its composition so as to maintain as high as possible a rate of replication. Sol Spiegelman is the inventor of 'unnatural selection'. He showed clearly that populations of RNA molecules evolve when replicated repeatedly by Q beta RNA polymerase under a chosen set of adverse reaction conditions. In the systems that he studied, the initial population was fairly homogeneous and much of the variation was created during the course of the experiment by mutation, that is, error of replication. The term 'unnatural selection' will be used loosely to describe evolution of nucleic acids or other replicatable polymers in vitro. The term 'Natural Selection' will be reserved for the evolution of living organisms and their viruses. Natural Section usually involves the coevolution of nucleic acids and proteins, while 'unnatural selection', as practiced so far, allows replicating nucleic acids to evolve but hold constant the enzymes that catalyze replication. It is widely believed that biology based on DNA, RNA, and proteins was preceded by the biology of an 'RNA world' in which enzymes were composed of RNA alone. The origin of RNA replication is thus the central puzzle of the origins of life. Consequently, RNA-catalyzed RNA replication is presently one of the main goals of experimental work on unnatural selection. However, there is also a more distant goal, namely, to achieve replication and selection in systems unrelated to RNA. These different systems are discussed in this article.
Increased human spaceflight operations utilize oxygen concentrations that are frequently varied with use of concentrations up to 100 percent oxygen. Even after exiting a higher percentage oxygen environment, high oxygen concentrations can still be maintained due to material saturation and oxygen entrapment between barrier materials. This paper examines the material flammability concerns that arise from changing oxygen environments during spaceflight operations. We examine the time required for common spacecraft and spacesuit materials exposed to oxygen to return to reduced ignitability and flammability once removed from the increased concentration. Various common spacecraft materials were considered: spacecraft cabin environment foams, Extra Vehicular Mobility Unit materials and foams, Advanced Crew Escape Suit materials, and other materials of interest such as Cotton, Nomex^ HT90-40, and Tiburon Surgical Drape. This paper presents calculated diffusion coefficients derived from experimentally obtained oxygen transmission rates for the tested materials and the analytically derived times necessary for reduced flammability to be achieved based on NASA flammability criteria. Oxygen material saturation and entrapment scenarios are examined. Experimental verification data on oxygen diffusion in saturation scenarios are also presented and discussed. We examine how to use obtained data to address flammability concerns during operational planning to reduce the likelihood of fires while improving efficiency for procedures.
The effect of radiative feedback from the gas phase in micro-gravity combustion processes has been of increasing concern because of the implications in the selection and evaluation of appropriate fire suppressants. The use of CO2, an optically thick gas in the infrared region of the electromagnetic spectrum, has garnered widespread acceptance as an effective fire suppressant for most ground based applications. Since buoyant forces often dominate the flow field in 1-g environments the temperature field between the flame front and the fuel surface is not significantly affected by gas phase radiative absorption and re-emission as these hot gases are quickly swept downstream. However, in reduced gravity environments where buoyant-driven convective flows are negligible and where low-speed forced convective flows may be present at levels where gas phase radiation becomes important, then changes in environment that enhance gas phase radiative effects need to be better understood. This is particularly true in assessments of flammability limits and selection of appropriate fire suppressants for future space applications. In recognition of this, a ground-based investigation has been established that uses a droplet combustion configuration to systematically study the effects of enhanced gas phase radiation on droplet burn rates, flame structure, and radiative output from the flame zone.
A goal of the aerospace industry is to reduce the cost of space transportation by a significant within the next decade. The present cost of launching a space transportation system which includes propulsion system, vehicle, carrier, and payload integrated together to form a system, encompasses much more than the design of the propulsion system and vehicle. The total cost includes the recurring cost of the process of integrating carriers, and payloads into the vehicle for each flight. The recurring cost of the integration of carrier/payloads systems is driven by the interaction of the vehicle. If the interaction can be well characterized and made to be very predictable for a range of payloads, or if it can be minimized then the cost of integrating a payload can be reduced significantly from today's levels. The Space Shuttle is very interactive with the payload. The interaction has been well characterized through finite element modeling and is reasonably predictable for a specific payload. Experience has shown, however, that the interaction is very manifest dependent, and small changes in one portion of a payload complement can change the interaction significantly in another portion. That is the affects of one on the other are such that if one or the other is changed slightly the environment at the interfaces can change significantly. To date the Shuttle has made in excess of one hundred flights. For each of these flights several iterations of dynamic analyses have been required in the development of each vehicle/carrier/payload system. The iterative analyses are needed because of the sensitivity of the interaction of the launch vehicle to the attached carrier/payload. The Multi Purpose Logistics Module (MPLM) is a carrier designed for flight in the Space Shuttle carrying a wide variation of cargo, supplies, and experiments to and from Space Station. Its integration process provides a unique area for improvement in the template in use today for transporting items to space. Discussion of the present MPLM integration analysis requirements and possible areas for improvement of the process are provided in the subject paper. Special emphasis is placed on reduction of requirements based on similarity of items being transported, and the use of carrier hardware for protection of the Shuttle Orbiter.