Objective:In this article, we investigated the effects of external human-machine interfaces (eHMIs) on pedestrians’ crossing intentions.Background:Literature suggests that the safety (i.e., not crossing when unsafe) and efficiency (i.e., crossing when safe) of pedestrians’ interactions with automated vehicles could increase if automated vehicles display their intention via an eHMI.Methods:Twenty-eight participants experienced an urban road environment from a pedestrian’s perspective using a head-mounted display. The behavior of approaching vehicles (yielding, nonyielding), vehicle size (small, medium, large), eHMI type (1. baseline without eHMI, 2. front brake lights, 3. Knightrider animation, 4. smiley, 5. text [WALK]), and eHMI timing (early, intermediate, late) were varied. For yielding vehicles, the eHMI changed from a nonyielding to a yielding state, and for nonyielding vehicles, the eHMI remained in its nonyielding state. Participants continuously indicated whether they felt safe to cross using a handheld button, and “feel-safe” percentages were calculated.Results:For yielding vehicles, the feel-safe percentages were higher for the front brake lights, Knightrider, smiley, and text, as compared with baseline. For nonyielding vehicles, the feel-safe percentages were equivalent regardless of the presence or type of eHMI, but larger vehicles yielded lower feel-safe percentages. The Text eHMI appeared to require no learning, contrary to the three other eHMIs.Conclusion:An eHMI increases the efficiency of pedestrian-AV interactions, and a textual display is regarded as the least ambiguous.Application:This research supports the development of automated vehicles that communicate with other road users.

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Both endogenous processes and exogenous physical and chemical sources generate deoxyribonucleic acid (DNA) damage in the nucleus and organelles of living cells. To prevent deleterious effects, damage is balanced by repair pathways. DNA repair was first documented for the nuclear compartment but evidence was subsequently extended to the organelles. Mitochondria and chloroplasts possess their own repair processes. These share a number of factors with the nucleus but also rely on original mechanisms. Base excision repair remains the best characterized. Repair is organized with the other DNA metabolism pathways in the organelle membrane-associated nucleoids. DNA repair in mitochondria is a regulated, stress-responsive process. Organelle genomes do not encode DNA repair enzymes and translocation of nuclear-encoded repair proteins from the cytosol seems to be a major control mechanism. Finally, changes in the fidelity and efficiency of mitochondrial DNA repair are likely to be involved in DNA damage accumulation, disease and aging. The present review successively addresses these different issues.

Group II introns are large catalytic RNAs that are ancestrally related to nuclear spliceosomal introns. Sequences corresponding to group II RNAs are found in many prokaryotes and are particularly prevalent within plants organellar genomes. Proteins encoded within the introns themselves (maturases) facilitate the splicing of their own host pre-RNAs. Mitochondrial introns in plants have diverged considerably in sequence and have lost their maturases. In angiosperms, only a single maturase has been retained in the mitochondrial DNA: the matR gene found within NADH dehydrogenase 1 (nad1) intron 4. Its conservation across land plants and RNA editing events, which restore conserved amino acids, indicates that matR encodes a functional protein. However, the biological role of MatR remains unclear. Here, we performed an in vivo investigation of the roles of MatR in Brassicaceae. Directed knockdown of matR expression via synthetically designed ribozymes altered the processing of various introns, including nad1 i4. Pull-down experiments further indicated that MatR is associated with nad1 i4 and several other intron-containing pre-mRNAs. MatR may thus represent an intermediate link in the gradual evolutionary transition from the intron-specific maturases in bacteria into their versatile spliceosomal descendants in the nucleus. The similarity between maturases and the core spliceosomal Prp8 protein further supports this intriguing theory.

Plant mitochondria are remarkable with respect to their content in foreign, alien and plasmid-like DNA, raising the question of the transfer of this information into the organelles. We demonstrate the existence of an active, transmembrane potential-dependent mechanism of DNA uptake into plant mitochondria. The process is restricted to double-strand DNA, but has no obvious sequence speci®city. It is most ef®cient with linear fragments up to a few kilobase pairs. When containing appropriate information, imported sequences are transcribed within the organelles. The uptake likely involves the voltage-dependent anion channel and the adenine nucleotide translocator, i.e. the core components of the mitochondrial permeability transition pore complex in animal cells, but it does not rely on known mitochondrial membrane permeabilization processes. We conclude that DNA import into plant mitochondria might represent a physiological phenomenon with some functional relevance.

The mitochondria of flowering plants have considerably larger and more complex genomes than the mitochondria of animals or fungi, mostly due to recombination activities that modulate their genomic structures. These activities most probably participate in the repair of mitochondrial DNA (mtDNA) lesions by recombination-dependent processes. Rare ectopic recombination across short repeats generates new genomic configurations that contribute to mtDNA heteroplasmy, which drives rapid evolution of the sequence organization of plant mtDNAs. We found that Arabidopsis thaliana RECG1, an ortholog of the bacterial RecG translocase, is an organellar protein with multiple roles in mtDNA maintenance. RECG1 targets to mitochondria and plastids and can complement a bacterial recG mutant that shows defects in repair and replication control. Characterization of Arabidopsis recG1 mutants showed that RECG1 is required for recombination-dependent repair and for suppression of ectopic recombination in mitochondria, most likely because of its role in recovery of stalled replication forks. The analysis of alternative mitotypes present in a recG1 line and of their segregation following backcross allowed us to build a model to explain how a new stable mtDNA configuration, compatible with normal plant development, can be generated by stoichiometric shift.

Replication of the Carnation Italian ringspot virus genomic RNA in plant cells occurs in multivesicular bodies which develop from the mitochondrial outer membrane during infection. ORF1 in the viral genome encodes a 36-kDa protein, while ORF2 codes for the 95-kDa replicase by readthrough of the ORF1 stop codon. We have shown previously that the N-terminal part of ORF1 contains the information leading to vesiculation of mitochondria and that the 36-kDa protein localizes to mitochondria. Using infection, in vivo expression of green fluorescent protein fusions in plant and yeast cells, and in vitro mitochondrial integration assays, we demonstrate here that both the 36-kDa protein and the complete replicase are targeted to mitochondria and anchor to the outer membrane with the N terminus and C terminus on the cytosolic side. Analysis of deletion mutants indicated that the anchor sequence is likely to correspond approximately to amino acids 84 to 196, containing two transmembrane domains. No evidence for a matrix-targeting presequence was found, and the data suggest that membrane insertion of the viral proteins is mediated by an import receptor-independent signal-anchor mechanism relying on the two transmembrane segments and multiple recognition signals present in the N-terminal part of ORF1.The genomes of positive-stranded RNA viruses from a number of supergroups are replicated in association with intracellular membranes (3). Depending on the virus, a variety of membrane systems can be concerned in infected cells, including the plasma membrane, endoplasmic reticulum, vacuole, chloroplasts, mitochondria, peroxisomes, and lysosomes (19,22,32,38,42,47). Little is known about the molecular mechanisms supporting these virus-cell interactions, which are critical for the development of infection. Membrane interaction with both viral proteins and host-encoded factors (50) has been proposed. Viral RNA replication is often associated with infection-specific membrane proliferation and/or vesiculation.Plant infection with Carnation Italian ringspot virus (CIRV), a member of the genus Tombusvirus in the family Tombusviridae, triggers the development of conspicuous membrane bodies from modified mitochondria (11). The CIRV genome is composed of a monopartite, positive-sense RNA genome of 4,760 nucleotides (GenBank accession number X85215). It is not polyadenylated, lacks a 5Ј cap structure, and contains five functional open reading frames (ORFs) (36) (Fig. 1). The 5Ј-most ORF (ORF1) in the CIRV genome encodes a 36-kDa protein (36K protein), while ORF2 codes for a 95-kDa protein (95K), which is expressed by readthrough of the amber stop codon of ORF1. Readthrough occurs at a frequency of about 10% in plant cells. Both pre-and complete readthrough products are essential for viral replication and were shown to contain the eight conserved motifs (PI to PVIII) of RNAdependent RNA polymerases of supergroup II of the positivestranded RNA viruses (16,40). ORF3 encodes the coat protein of 41 kDa, and the two nested ORFs 4 and 5 code for 21...

Respiration, a fundamental process in mammalian cells, requires two genomes, those of the nucleus and the mitochondrion (mtDNA). Mutations of mtDNA are being increasingly recognized in disease and may play an important role in the ageing process. Accepting the vital role of mtDNA gene products, our limited knowledge concerning the details of mitochondrial gene expression is surprising. This is, in part, due to our inability to transfect mitochondria and to manipulate their genome. There have been claims of successful DNA import into isolated organelles, but most reports lacked evidence of expression and no method has furthered our understanding of gene expression. Here, we report that mammalian mitochondria possess a natural competence for DNA import. Using five functional assays, we show imported DNA can act as templates for DNA synthesis or promoter-driven transcription, with the resultant polycistronic RNA being processed (5' and 3') and excised mt-tRNA matured. Exploiting this natural competence will allow us to explore mitochondrial gene expression in organello and provides the potential for mitochondrial transfection in vivo.