scholarly journals Fidelity of a Bacterial DNA Polymerase in Microgravity, a Model for Human Health in Space

Author(s):  
Aaron H Rosenstein ◽  
Virginia K Walker

Long-term space missions will expose crew members, their cells as well as their microbiomes to prolonged periods of microgravity and ionizing radiation, environmental stressors for which almost no earth-based organisms have evolved to survive. Despite the importance of maintaining genomic integrity, the impact of these stresses on DNA polymerase-mediated replication and repair has not been fully explored. DNA polymerase fidelity and replication rates were assayed under conditions of microgravity generated by parabolic flight and compared to earth-like gravity. Upon commencement of a parabolic arc, primed synthetic single-stranded DNA was used as a template for one of two enzymes (Klenow fragment exonuclease+/−; with and without proofreading exonuclease activity, respectively) and were quenched immediately following the 20 s microgravitational period. DNA polymerase error rates were determined with an algorithm developed to identify experimental mutations. In microgravity Klenow exonuclease+ showed a median 1.1-fold per-base decrease in polymerization fidelity for base substitutions when compared to earth-like gravity (p = 0.02), but in the absence of proofreading activity, a 2.4-fold decrease was observed (p = 1.98 × 10−11). Similarly, 1.1-fold and 1.5-fold increases in deletion frequencies in the presence or absence of exonuclease activity (p = 1.51 × 10−7 and p = 8.74 × 10−13), respectively, were observed in microgravity compared to controls. The development of this flexible semi-autonomous payload system coupled with genetic and bioinformatic approaches serves as a proof-of-concept for future space health research.

1994 ◽  
Vol 302 (2) ◽  
pp. 567-571 ◽  
Author(s):  
P Hentosh ◽  
P Grippo

2-Chloro-2′-deoxyadenosine triphosphate, a purine nucleotide analogue and potent antileukaemic agent, was incorporated into double-stranded 36-mers in place of dATP to investigate the effects of 2-chloroadenine (ClAde) on DNA polymerase-associated 3′-->5′ exonuclease activity. ClAde residues within one strand of duplex DNA did not inhibit exonuclease activity; on the contrary, ClAde-containing minus strands were digested to a greater extent than was control DNA in the absence of deoxyribonucleoside triphosphates by Escherichia coli Klenow fragment, yeast DNA polymerase II and T4 DNA polymerase. After a 30 min incubation with 5 units of Klenow fragment, approximately 65% of control DNA remained in DNA fragments of 26 bases or larger compared with only approximately 25% of ClAde-substituted substrates. Unsubstituted plus strands opposite a ClAde-containing strand were likewise digested more quickly by 3′-->5′ exonuclease, but only in the vicinity of the ClAde sites. Approx. 63% of the plus strands from ClAde-containing oligomers were less than 24 bases in length after a 25 min digestion period with Klenow fragment compared with only approximately 32% of control DNA. Such results indicate that, unlike other base modifications such as pyrimidine dimers, methoxy psoralen adducts and certain nucleoside analogues, all of which inhibit or decrease the rate of strand degradation by 3′-->5′ exonucleases, incorporated ClAde enhances strand degradation of duplex DNA.


2019 ◽  
Author(s):  
Nicolas Meirhaeghe ◽  
Virginie Bayet ◽  
Pierre-Vincent Paubel ◽  
Claudine Mélan

ABSTRACTUnderstanding the impact of microgravity on human cognitive performance is crucial to guarantee the safety and success of future long-term manned missions. The effects of weightlessness on key mental processes such as spatial abilities are in particular not fully characterized. In this study, we examine the influence of microgravity on perspective-taking abilities—a type of mental operation especially relevant in the context of collaborative teamwork between ‘free-floating’ astronauts. Twelve subjects performed a cooperative task in virtual-reality under both normal and short-term microgravity conditions during a parabolic flight. The task involved various degrees of mental transformations, and required subjects to perform actions instructed by a fellow astronaut aboard a virtual spacecraft. The experimental design allowed us to control for nuisance variables, training effects, and non-gravity related factors of parabolic flights. Overall, our results indicated that microgravity has a facilitatory effect on perspective-taking abilities. Notably, this facilitation was selective to conditions requiring subjects to rotate their perspective around their line of sight, i.e., for mental rotations in the frontal plane. Moreover, microgravity affected subjects differently depending on their visual field dependence, as determined via a classic rod-and-frame test. Specifically, improvement in performance was more pronounced in field-independent subjects. Together, these results shed light on a long standing debate about the impact of microgravity on egocentric mental imagery, and have direct operational consequences for future long-term missions.


2018 ◽  
Vol 31 (5) ◽  
pp. 373-389 ◽  
Author(s):  
Simon P. Landry ◽  
François Champoux

Long-term musical training is an enriched multisensory training environment that can alter uni- and multisensory substrates and abilities. Amongst these altered abilities are faster reaction times for simple and complex sensory tasks. The crossed arm temporal-order judgement (TOJ) task is a complex tactile task in which TOJ error rate increases when arms are crossed. Reaction times (RTs) for this task are typically proportionate to the difficulty of the stimulus onset asynchrony (SOA) and increase more when the arms are crossed than when uncrossed. The objective of this study was to study the impact of musical training on RTs and accuracy for the crossed arm TOJ task. Seventeen musicians and 20 controls were tested. Musicians had significantly faster RTs for all crossed arm conditions and half of the uncrossed conditions. However, musicians had significantly more TOJ errors for the crossed posture. We speculate that faster musician TOJ RTs leave little time to consolidate conflicting internal and external task-related information when crossing the arms, leading to increased incorrect responses. These results provide novel insights on the potential mechanisms underlying the increased TOJ error rates when arms are crossed. Moreover, they add to the growing literature of altered sensory ability in musicians and propose an unexpected consequence of faster reaction times.


2011 ◽  
Vol 70 (1) ◽  
pp. 5-11 ◽  
Author(s):  
Beat Meier ◽  
Anja König ◽  
Samuel Parak ◽  
Katharina Henke

This study investigates the impact of thought suppression over a 1-week interval. In two experiments with 80 university students each, we used the think/no-think paradigm in which participants initially learn a list of word pairs (cue-target associations). Then they were presented with some of the cue words again and should either respond with the target word or avoid thinking about it. In the final test phase, their memory for the initially learned cue-target pairs was tested. In Experiment 1, type of memory test was manipulated (i.e., direct vs. indirect). In Experiment 2, type of no-think instructions was manipulated (i.e., suppress vs. substitute). Overall, our results showed poorer memory for no-think and control items compared to think items across all experiments and conditions. Critically, however, more no-think than control items were remembered after the 1-week interval in the direct, but not in the indirect test (Experiment 1) and with thought suppression, but not thought substitution instructions (Experiment 2). We suggest that during thought suppression a brief reactivation of the learned association may lead to reconsolidation of the memory trace and hence to better retrieval of suppressed than control items in the long term.


2003 ◽  
Author(s):  
Teresa Garate-Serafini ◽  
Jose Mendez ◽  
Patty Arriaga ◽  
Larry Labiak ◽  
Carol Reynolds

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