Humor and the willingness to buy healthy food posted on Instagram.

Humorous messages (not derogatory jokes) related to obesity seem to be retweeted frequently. Potentially, such humor could be included in viral public health campaigns to combat obesity, but would jokes actually increase the likelihood of purchasing healthy foods advertised on social media? 411 participants were asked to test two soon-to-be-introduced features (e.g., repost button) in Instagram on their phones. Participants scrolled through a series of posts about various topics ranging from architecture to beauty products. A healthy food post, preceded by one of four joke types (two containing the word "fat"), was embedded at the top, middle or bottom position of the Instagram page. After participants scrolled through the page, perhaps reposting some posts, they were presented with the healthy food product featured in the post and were asked to indicate a purchase probability (0-100), as well as whether they recognized the food product from the post (yes/no). At the end of the study, the individual jokes were rated as "funny/not funny" and "positive/negative". If the food product was recognized, the joke type played a role. In particular, jokes containing the word "fat" had a negative effect on the purchase probability. However, if the food product was not recognized, only its position on the page mattered. The purchase probability was highest if the product was placed at the top. Social media, criticized for promoting unhealthy food consumption, especially among children, can therefore also be used to address issues such as the global obesity epidemic.

Online Food Choices: When Do "Recommended By" Labels Work?

Understanding digital menu choices in limited-option environments, such as university cafés, is crucial for promoting healthier and more sustainable food choices. We are, therefore, looking at two possible interventions or nudges-recommendation and position-and how they interact with, for example, price. In the first smartphone-based study (N = 517), participants were presented with two menu options, while the factors "recommendation", "position", and "price" were manipulated. We only found effects in relation to the choice of the more popular menu option. Specifically, when the popular meal was the expensive option, the recommendation had a negative effect on choice, but not when the popular meal was the cheaper option. The aim of the second smartphone-based study (N = 916) was to shed more light on the role of popularity or personal preference in relation to recommendations. We manipulated the differences in personal preference (small or large) using a ranking task presented before the menu choice. In Study 2, the interaction effect between recommendation and price for the more popular menu option could not be replicated. Instead, we found that the greater the difference in preference, the less pronounced the price effect was. Overall, some effects of the recommendations have been identified, but further research is needed to clarify the exact circumstances under which they arise.

Do Pictograms on Medication Packages Cause People to Consult Package Inserts Less Often? If so, With What Consequences?

Overall, pharmaceutical pictograms seem to improve medication adherence. However, little is known about how warning pictograms (e.g., "do not drive after taking") on medication packages influence patients' information-seeking strategies such as consulting the package insert (PI) to determine other features such as the correct dosage. In this online study, participants (358 students) were presented with three fictitious scenarios (e.g., headache after alcohol consumption; factor scenario) in which medication use would be contraindicated. Each scenario was accompanied by a visual presentation of a medication package that could contain three possible pictogram selections or arrangements (factor warning); some arrangements contained pictograms relevant to the situation represented by the scenario, while others did not. Participants had to decide which dosage of the represented medication they were allowed to take in the given scenario. In making this decision, they could consult the PI or not. Overall, in two out of the three scenarios (driving and pregnancy), medication packages with relevant pictograms resulted in fewer PI consultations but led to more correct dosage decisions ("no pill") than packages with irrelevant pictograms. Pictograms generally played no role in either the review of the PI consultation or dosage decisions in the alcohol scenario. Providing warning-relevant pictograms on medication packages can help people know when they should not take medication even without reading the PI.

A New Bacterial Adenosine-Derived Nucleoside as an Example of RNA Modification Damage.

The fields of RNA modification and RNA damage both exhibit a plethora of non-canonical nucleoside structures. While RNA modifications have evolved to improve RNA function, the term RNA damage implies detrimental effects. Based on stable isotope labelling and mass spectrometry, we report the identification and characterisation of 2-methylthio-1,N6-ethenoadenosine (ms2 ϵA), which is related to 1,N6-ethenoadenine, a lesion resulting from exposure of nucleic acids to alkylating chemicals in vivo. In contrast, a sophisticated isoprene labelling scheme revealed that ms2 ϵA biogenesis involves cleavage of a prenyl moiety in the known transfer RNA (tRNA) modification 2-methylthio-N6-isopentenyladenosine (ms2 i6 A). The relative abundance of ms2 ϵA in tRNAs from translating ribosomes suggests reduced function in comparison to its parent RNA modification, establishing the nature of the new structure in a newly perceived overlap of the two previously separate fields, namely an RNA modification damage.

Landscape of Post-Transcriptional tRNA Modifications in Streptomyces albidoflavus J1074 as Portrayed by Mass Spectrometry and Genomic Data Mining.

Actinobacterial genus Streptomyces (streptomycetes) represents one of the largest cultivable group of bacteria famous for their ability to produce valuable specialized (secondary) metabolites. Regulation of secondary metabolic pathways inextricably couples the latter to essential cellular processes that determine levels of amino acids, carbohydrates, phosphate, etc. Post-transcriptional tRNA modifications remain one of the least studied aspects of streptomycete physiology, albeit a few of them were recently shown to impact antibiotic production. In this study, we describe the diversity of post-transcriptional tRNA modifications in model strain Streptomyces albus (albidoflavus) J1074 by combining mass spectrometry and genomic data. Our results show that J1074 can produce more chemically distinct tRNA modifications than previously thought. An in silico approach identified orthologs for enzymes governing most of the identified tRNA modifications. Yet, genetic control of certain modifications remained elusive, suggesting early divergence of tRNA modification pathways in Streptomyces from the better studied model bacteria, such as Escherichia coli and Bacillus subtilis. As a first point in case, our data point to the presence of a non-canonical MiaE enzyme performing hydroxylation of prenylated adenosines. A further finding concerns the methylthiotransferase MiaB, which requires previous modification of adenosines by MiaA to i6A for thiomethylation to ms2i6A. We show here that the J1074 ortholog, when overexpressed, yields ms2A in a ΔmiaA background. Our results set the working ground for and justify a more detailed studies of biological significance of tRNA modification pathways in streptomycetes. IMPORTANCE Post-transcriptional tRNA modifications (PTTMs) play an important role in maturation and functionality of tRNAs. Little is known about tRNA modifications in the antibiotic-producing actinobacterial genus Streptomyces, even though peculiar tRNA-based regulatory mechanisms operate in this taxon. We provide a first detailed description of the chemical diversity of PTTMs in the model species, S. albidoflavus J1074, and identify most plausible genes for these PTTMs. Some of the PTTMs are described for the first time for Streptomyces. Production of certain PTTMs in J1074 appears to depend on enzymes that show no sequence similarity to known PTTM enzymes from model species. Our findings are of relevance for interrogation of genetic basis of PTTMs in pathogenic actinobacteria, such as M. tuberculosis.

Functional integration of a semi-synthetic azido-queuosine derivative into translation and a tRNA modification circuit.

Substitution of the queuine nucleobase precursor preQ1 by an azide-containing derivative (azido-propyl-preQ1) led to incorporation of this clickable chemical entity into tRNA via transglycosylation in vitro as well as in vivo in Escherichia coli, Schizosaccharomyces pombe and human cells. The resulting semi-synthetic RNA modification, here termed Q-L1, was present in tRNAs on actively translating ribosomes, indicating functional integration into aminoacylation and recruitment to the ribosome. The azide moiety of Q-L1 facilitates analytics via click conjugation of a fluorescent dye, or of biotin for affinity purification. Combining the latter with RNAseq showed that TGT maintained its native tRNA substrate specificity in S. pombe cells. The semi-synthetic tRNA modification Q-L1 was also functional in tRNA maturation, in effectively replacing the natural queuosine in its stimulation of further modification of tRNAAsp with 5-methylcytosine at position 38 by the tRNA methyltransferase Dnmt2 in S. pombe. This is the first demonstrated in vivo integration of a synthetic moiety into an RNA modification circuit, where one RNA modification stimulates another. In summary, the scarcity of queuosinylation sites in cellular RNA, makes our synthetic q/Q system a 'minimally invasive' system for placement of a non-natural, clickable nucleobase within the total cellular RNA.

Well-designed medical pictograms accelerate search.

Two types of newly designed pharmaceutical pictograms (with and without context) were compared with an existing type of certified pictograms regarding their search efficiency. Each of the 30 participants had to search a total of 1'090 "fictitious" medical shelves for a certain box defined by the amount and type of medical instructions given (memory size) and presented among a variable number of other boxes (set size). The boxes contained the different types of pictograms mentioned above. Calculated factorial analyses on reaction time data, among others, showed that the two newly designed pictogram types make search more efficient compared to existing types of pictograms (i.e., flatter reaction time x set size slopes). Furthermore, regardless of the type of pictogram, this set size effect became more pronounced with larger memory sizes. Overall, the newly designed pictograms need fewer attentional resources and therefore might help to increase patient adherence.

Can carbon labels encourage green food choices?

Introduction: A conventionally grown kiwi from Spain or an organic pineapple from Ghana? Which is the more environmentally friendly option? Given that the production and distribution of food is responsible for about a quarter of our CO2e emissions and thus plays a role in climate change the answer to such questions and, accordingly, making the right food product choices is crucial. The problem, however, is that it is difficult for consumers to calculate the CO2e value of food as it depends on several specifications such as the type of food, origin, etc. Could carbon labeling of food circumvent this problem and help consumers make more environmentally friendly choices? Methods: In an online experiment, 402 participants had to choose 20 food products from a fictitious online shop. The participants were randomly assigned to either one of three food labeling conditions (Star Rating, Green Foot, and Traffic Light Label, short TLL) or the control condition. Results: The labeling conditions resulted in lower overall CO2e emissions, the purchase of more green food products and fewer red food products than in the control condition. The TLL outperformed the other two labels and was also the most accepted. Discussion: The carbon TLL is therefore a promising intervention to help consumers to not only choose more environmentally friendly foods, but also make a significant contribution to the fight against climate change.

Mutant-selective degradation by BRAF-targeting PROTACs.

Over 300 BRAF missense mutations have been identified in patients, yet currently approved drugs target V600 mutants alone. Moreover, acquired resistance inevitably emerges, primarily due to RAF lesions that prevent inhibition of BRAF V600 with current treatments. Therefore, there is a need for new therapies that target other mechanisms of activated BRAF. In this study, we use the Proteolysis Targeting Chimera (PROTAC) technology, which promotes ubiquitination and degradation of neo-substrates, to address the limitations of BRAF inhibitor-based therapies. Using vemurafenib-based PROTACs, we achieve low  nanomolar degradation of all classes of BRAF mutants, but spare degradation of WT RAF family members. Our lead PROTAC outperforms vemurafenib in inhibiting cancer cell growth and shows in vivo efficacy in a Class 2 BRAF xenograft model. Mechanistic studies reveal that BRAFWT is spared due to weak ternary complex formation in cells owing to its quiescent inactivated conformation, and activation of BRAFWT sensitizes it to degradation. This study highlights the degree of selectivity achievable with degradation-based approaches by targeting mutant BRAF-driven cancers while sparing BRAFWT, providing an anti-tumor drug modality that expands the therapeutic window.

Bacterial tRNA 2'-O-methylation is dynamically regulated under stress conditions and modulates innate immune response.

RNA modifications are a well-recognized way of gene expression regulation at the post-transcriptional level. Despite the importance of this level of regulation, current knowledge on modulation of tRNA modification status in response to stress conditions is far from being complete. While it is widely accepted that tRNA modifications are rather dynamic, such variations are mostly assessed in terms of total tRNA, with only a few instances where changes could be traced to single isoacceptor species. Using Escherichia coli as a model system, we explored stress-induced modulation of 2'-O-methylations in tRNAs by RiboMethSeq. This analysis and orthogonal analytical measurements by LC-MS show substantial, but not uniform, increase of the Gm18 level in selected tRNAs under mild bacteriostatic antibiotic stress, while other Nm modifications remain relatively constant. The absence of Gm18 modification in tRNAs leads to moderate alterations in E. coli mRNA transcriptome, but does not affect polysomal association of mRNAs. Interestingly, the subset of motility/chemiotaxis genes is significantly overexpressed in ΔTrmH mutant, this corroborates with increased swarming motility of the mutant strain. The stress-induced increase of tRNA Gm18 level, in turn, reduced immunostimulation properties of bacterial tRNAs, which is concordant with the previous observation that Gm18 is a suppressor of Toll-like receptor 7 (TLR7)-mediated interferon release. This documents an effect of stress induced modulation of tRNA modification that acts outside protein translation.

Effects of Bag Mask Ventilation and Advanced Airway Management on Adherence to Ventilation Recommendations and Chest Compression Fraction: A Prospective Randomized Simulator-Based Trial.

The role of advanced airway management (AAM) in cardiopulmonary resuscitation (CPR) is currently debated as observational studies reported better outcomes after bag-mask ventilation (BMV), and the only prospective randomized trial was inconclusive. Adherence to CPR guidelines ventilation recommendations is unknown and difficult to assess in clinical trials. This study compared AAM and BMV with regard to adherence to ventilation recommendations and chest compression fractions in simulated cardiac arrests. A total of 154 teams of 3-4 physicians were randomized to perform CPR with resuscitation equipment restricting airway management to BMV only or equipment allowing for all forms of AAM. BMV teams ventilated 6 ± 6/min and AAM teams 19 ± 8/min (range 3-42/min; p < 0.0001 vs. BMV). 68/78 BMV teams and 23/71 AAM teams adhered to the ventilation recommendations (p < 0.0001). BMV teams had lower compression fractions than AAM teams (78 ± 7% vs. 86 ± 6%, p < 0.0001) resulting entirely from higher no-flow times for ventilation (9 ± 4% vs. 3 ± 3 %; p < 0.0001). Compared to BMV, AAM leads to significant hyperventilation and lower adherence to ventilation recommendations but favourable compression fractions. The cumulative effect of deviations from ventilation recommendations has the potential to blur findings in clinical trials.

Regulation of human inducible nitric oxide synthase expression by an upstream open reading frame.

The human inducible nitric oxide synthase (iNOS) gene contains an upstream open reading frame (uORF) in its 5'-untranslated region (5'-UTR) implying a translational regulation of iNOS expression. Transfection experiments in human DLD-1 cells revealed that the uORF although translatable seems not to inhibit the translation start at the bona fide ATG. Our data clearly show that human iNOS translation is cap-dependent and that the 5'-UTR of the iNOS mRNA contains no internal ribosome entry site. Translation of the bona fide coding sequence is most likely mediated by a leaky scanning mechanism. The 5'-UTR is encoded by exon 1 and exon 2 of the iNOS gene with the uORF stop codon located in front of the first intron indicating an involvement of the nonsense mediated RNA decay (NMD) in iNOS regulation. SiRNA-mediated down-regulation of Upf1 resulted in enhanced endogenous cytokine iNOS expression in human DLD-1 cells. Transfection of constructs containing iNOS exon 1, intron 1 and exon 2 in front of a luciferase gene showed a clear effect of the mutation of the uORF-ATG on luciferase reportergene expression. Our data indicate that the uORF in the 5'-UTR sequence of human iNOS gene reduces its expression via the NMD mechanism.

Double stimuli-responsive polysaccharide block copolymers as green macrosurfactants for near-infrared photodynamic therapy.

The NIR absorbing photosensitizer phthalocyanine zinc (PC(Zn)) was stabilized in aqueous media as water-dispersible nanoparticles with a reduction- and pH-responsive full polysaccharide block copolymer. A cellular uptake and also photo switchable intracellular activity of the cargo upon irradiation at wavelengths in the near infrared region were shown. The block copolymer was synthesized by applying a copper-free click strategy based on a thiol exchange reaction, creating an amphiphilic double-stimuli-responsive mixed disulfide. The dual-sensitive polysaccharide micelles represent a non-toxic and biodegradable green macrosurfactant for the delivery of phthalocyanine zinc. By encapsulation into micellar nanoparticles, the bioavailability of PC(Zn) increased significantly, enabling smart photodynamic therapy for future applications in cancer-related diseases.

Influence of activated charcoal amendment to contaminated soil on dieldrin and nutrient uptake by cucumbers.

Activated charcoal (AC) amendments have been suggested as a promising, cost-effective method to immobilize organic contaminants in soil. We performed pot experiments over two years with cucumber (Cucumis sativus L.) grown in agricultural soil with 0.07 mg kg(-1) of weathered dieldrin and 0, 200, 400, and 800 mg AC per kg soil. Dieldrin fresh weight concentrations in cucumber fruits were significantly reduced from 0.012 to an average of 0.004 mg kg(-1), and total uptake from 2 to 1 microg in the 800 mg kg(-1) AC treatment compared to the untreated soil. The treatment effects differed considerably between the two years, due to different meteorological conditions. AC soil treatments did neither affect the availability of nutrients to the cucumber plants nor their yield (total fruit wet weight per pot). Thus, some important prerequisites for the successful application of AC amendments to immobilize organic pollutants in agricultural soils can be considered fulfilled.