The spatio-temporal regulation of gene expression determines the fate and function of various cells and tissues and, as a consequence, the correct development and functioning of complex organisms. Certain mechanisms of gene activity regulation provide adequate cell responses to changes in environmental factors. Aside from gene expression disorders that lead to various pathologies, alterations of expression of particular genes were shown to significantly decrease or increase the lifespan in a wide range of organisms from yeast to human. Drosophila fruit fly is an ideal model system to explore mechanisms of longevity and aging due to low cost, easy handling and maintenance, large number of progeny per adult, short life cycle and lifespan, relatively low number of paralogous genes, high evolutionary conservation of epigenetic mechanisms and signalling pathways, and availability of a wide range of tools to modulate gene expression in vivo. Here, we focus on the organization of the evolutionarily conserved signaling pathways whose components significantly influence the aging process and on the interconnections of these pathways with gene expression regulation.

Road traffic accidents (RTAs) are an ever-existing threat to all road users. Automated vehicles (AVs; SAE Level 3-5) are developed in many countries. They are promoted with numerous benefits such as increased safety yielding less RTAs, less congestion, less greenhouse gas emissions, and the possibility of enabling non-driving related tasks (NDRTs). However, there has been no study which has investigated different NDRT conditions, while comparing participants who experienced a severe RTA in the past with those who experienced no RTA. Therefore, we conducted a driving simulator study (N = 53) and compared two NDRT conditions (i.e., auditory-speech (ASD) vs. heads-up display (HUD)) and an accident (26 participants) with a non-accident group (27; between-subjects design). Although our results did not reveal any interaction effect, and no group difference between the accident and the non-accident group on NDRT, take-over request (TOR), and driving performance, we uncovered for both groups better performances for the HUD condition, whereas a lower cognitive workload was reported for the ASD condition. Nevertheless, there was no difference for technology trust between the two conditions. Albeit we observed higher self-ratings of PTSD symptoms for the accident than for the non-accident group, there were no group differences on depression and psychological resilience self-ratings. We conclude that severe RTA experiences do not undermine NDRT, TOR, and driving performance in a SAE Level 3 driving simulator study, although PTSD symptoms after an RTA may affect the psychological wellbeing.

Autophagy is an evolutionarily conservative biological process in eukaryotes. Since the lysosomes were discovered by De Duve in the 1950s, autophagy has been studied for more than half a century and the mechanism of autophagy process has been discovered in many model organisms. In the rice blast fungus Magnaporthe oryzae, autophagy relative proteins are essential for appressorium formation, penetration, and invasive growth. The null mutants for the expression of autophagy gene homologs in M. oryzae lose their pathogenicity for infection of host plants. In this chapter, we provide some methods for monitoring autophagy process using physics and biochemistry assays in M. oryzae. Moreover, similar approaches can be used to monitor autophagy in other plant filamentous pathogenic fungi.

Crumb rubber (CR) manufactured from waste tires used in bitumen could improve bitumen performance and reduce environmental pollution. In this work, three different modification methods, pretreatment of the CR particles (microwave activation), warm mix additive (Sasobit), and trans-polyoctenamer (TOR) were used to improve the compatibility of CR with bitumen. Moreover, two other specimens, Sasobit and microwave activated and TOR and microwave activation were fabricated, and their performance was investigated. The softening point, elastic recovery, force ductility, rotational viscosity, temperature sweep, frequency sweep, and multiple stress creep and recovery (MSCR) tests were measured to evaluate the physical and rheological properties of rubberized bitumen. The results showed that TOR improved the physical properties of rubberized bitumen significantly but had a negative effect on the viscosity. Sasobit decreased the viscosity of rubberized bitumen considerably and improved the physical properties of rubberized bitumen moderately. Microwave treatment on CR had a negative effect on the high temperature performance and elastic recoverability of rubberized bitumen, however, attributing to the degradation and devulcanization effect of microwave on CR, the viscosity of rubberized bitumen was improved. From the results of composite modification, the influence of TOR on the performance of rubberized bitumen was more obvious than that of the microwave activation treatment. Moreover, the influence of Sasobit on its performance was less than that of the microwave activation treatment.

Iron is an essential micronutrient that participates as a cofactor in a broad range of metabolic processes including mitochondrial respiration, DNA replication, protein translation and lipid biosynthesis. Adaptation to iron deficiency requires the global reorganization of cellular metabolism directed to optimize iron utilization. The budding yeast Saccharomyces cerevisiae has been widely used to characterize the responses of eukaryotic microorganisms to iron depletion. In this report, we used a genomic approach to investigate the contribution of transcription rates to the modulation of mRNA levels during adaptation of yeast cells to iron starvation. We reveal that a decrease in the activity of all RNA polymerases contributes to the down-regulation of many mRNAs, tRNAs and rRNAs. Opposite to the general expression pattern, many genes including components of the iron deficiency response, the mitochondrial retrograde pathway and the general stress response display a remarkable increase in both transcription rates and mRNA levels upon iron limitation, whereas genes encoding ribosomal proteins or implicated in ribosome biogenesis exhibit a pronounced fall. This expression profile is consistent with an activation of the environmental stress response. The phosphorylation stage of multiple regulatory factors strongly suggests that the conserved nutrient signaling pathway TORC1 is inhibited during the progress of iron deficiency. These results suggest an intricate crosstalk between iron metabolism and the TORC1 pathway that should be considered in many disorders.

Aging is a complex phenomenon caused by the time-dependent loss of cellular homeodynamics and consequently of physiological organismal functions. This process is affected by both genetic and environmental (e.g., diet) factors, as well as by their constant interaction. Consistently, deregulation of nutrient sensing and signaling pathways is considered a hallmark of aging. Nutrigenomics is an emerging scientific discipline that studies changes induced by diet on the genome and thus it considers the intersection of three topics, namely health, diet, and genomics. Model organisms, such as the fruit fly Drosophila melanogaster, have been successfully used for in vivo modeling of higher metazoans aging and for nutrigenomic studies. Drosophila is a well-studied organism with sophisticated genetics and a fully annotated sequenced genome, in which ~ 75% of human disease-related genes have functional orthologs. Also, flies have organs/tissues that perform the equivalent functions of most mammalian organs, while discrete clusters of cells maintain insect carbohydrate homeostasis in a way similar to pancreatic cells. Herein, we discuss the mechanistic connections between nutrition and aging in Drosophila, and how this model organism can be used to study the effect of different diets (including natural products and/or their derivatives) on higher metazoans longevity.

Web browsers are among the most commonly used applications to access the web from any platform nowadays. With recent digital incidents involving breach of data, users are becoming more cognizant of the threat posed by malicious actors having access to personal data as well as vulnerable applications which may compromise their data. For this very reason, users are being offered privacy preserving solutions for trust maturity. The onion router (Tor) browser is one such application which not only ensures the privacy preservation goals but also provides promising anonymity. Due to this feature, majority of the users use Tor browser for normal use as well as malign activities. In order to validate the claims of Tor browser and help digital forensic investigators and researchers, we created different scenarios to forensically analyze the Tor browser privacy and anonymity. As a result of the findings, it can be concluded that the Tor browser leaves plethora of sensitive digital artifacts on host machine, which can be further used to compromise user data.

We have previously conducted the Sirolimus and Autophagy Inhibition in LAM (SAIL) trial, a phase 1 dose-escalation study of the combination of sirolimus and hydroxychloroquine in patients with lymphangioleiomyomatosis (LAM). The goal of the present study was to analyze sera from the SAIL trial to identify novel biomarkers that could shed light into disease pathogenesis and response to therapy.
We used the DiscoveryMAP platform from Rules Based Medicine to simultaneously measure 279 analytes in sera collected at each visit from subjects enrolled in the SAIL trial. We used longitudinal regression and pathway analysis to examine analyte rate of change and corresponding effect on lung function and to identify networks and potential nodes of interest.
A total of 222 analytes were included in the analysis. We identified 32 analytes that changed over the treatment period of the study. Pathway analysis revealed enrichment in cytokine-receptor interaction and mechanistic/mammalian target of rapamycin-related pathways, in addition to seemingly unrelated processes such as rheumatoid arthritis. Search Tool for the Retrieval of Interacting Genes/Proteins analysis identified two hubs centered around acetyl-CoA carboxylase alpha and beta and coagulation factor II. In addition, we identified vascular endothelial growth factor receptor-3 and CCL21 as molecules significantly associated with changes in FEV1 during the study period.
We performed a large-scale analyte study in sera of women with LAM and identified potential markers that could be linked to disease pathogenesis, lung injury, and therapeutic response. These data will enable future investigation into the specific roles of these molecules in LAM.
ClinicalTrials.gov; No. NCT01687179; URL: www.clinicaltrials.gov).

BACKGROUND: Termination of resuscitation guidelines for out-of-hospital cardiac arrest can identify patients in whom continuing resuscitation has little chance of success. This study examined the outcomes of patients transferred to hospital with ongoing CPR. It assessed outcomes for those who would have met the universal prehospital termination of resuscitation criteria (no shocks administered, unwitnessed by emergency medical services, no return of spontaneous circulation).
METHODS: A retrospective cohort study of consecutive adult patients who were transported to hospital with ongoing CPR was conducted at three hospitals in the West Midlands, UK between September 2016 and November 2017. Patient characteristics, interventions and response to treatment (ROSC, survival to discharge) were identified.
RESULTS: 227 (median age 69 years, 67.8% male) patients were identified. 89 (39.2%) met the universal prehospital termination of resuscitation criteria. Seven (3.1%) were identified with a potentially reversible cause of cardiac arrest. After hospital arrival, patients received few specialist interventions that were not available in the prehospital setting. Most (n = 210, 92.5%) died in the emergency department. 17 were admitted (14 to intensive care), of which 3 (1.3%) survived to hospital discharge. There were no survivors (0%) in those who met the criteria for universal prehospital termination of resuscitation.
CONCLUSIONS: Overall survival amongst patients transported to hospital with ongoing CPR was very poor. Application of the universal prehospital termination of resuscitation rule, in patients without obvious reversible causes of cardiac arrest, would have allowed resuscitation to have been discontinued at the scene for 39.2% of patients who did not survive.

Pathogen transmission by mosquitoes is tightly linked to blood feeding which, in turn, is required for egg development. Studies of these processes would greatly benefit from genetic methods, such as the binary Gal4/UAS system. The latter has been well established for model organisms, but its availability is limited for mosquitoes. The objective of this study was to develop the blood-meal-activated, gut-specific Gal4/UAS system for the yellow-fever mosquito Aedes aegypti and utilize it to investigate the regulation of gut-specific gene expression. A 1.1-kb, 5(\') upstream region of the carboxypeptidase A (CP) gene was used to genetically engineer the CP-Gal4 driver mosquito line. The CP-Gal4 specifically activated the Enhanced Green Fluorescent Protein (EGFP) reporter only after blood feeding in the gut of the CP-Gal4 > UAS-EGFP female Ae. aegypti. We used this system to study the regulation of CP gene expression. In vitro treatments with either amino acids (AAs) or insulin stimulated expression of the CP-Gal4 > UAS-EGFP transgene; no effect was observed with 20-hydroxyecdysone (20E) treatments. The transgene activation by AAs and insulin was blocked by rapamycin, the inhibitor of the Target-of-Rapamycin (TOR) kinase. RNA interference (RNAi) silence of the insulin receptor (IR) reduced the expression of the CP-Gal4 > UAS-EGFP transgene. Thus, in vitro and in vivo experiments have revealed that insulin and TOR pathways control expression of the digestive enzyme CP. In contrast, 20E, the major regulator of post-blood-meal vitellogenic events in female mosquitoes, has no role in regulating the expression of this gene. This novel CP-Gal4/UAS system permits functional testing of midgut-specific genes that are involved in blood digestion and interaction with pathogens in Ae. aegypti mosquitoes.