'Black Out Rage Gallon' (aka borg): An investigation of a risky drinking trend on TikTok.

ISSUE ADDRESSED: A 'Black Out Rage Gallon' (borg) is a customised, individual alcoholic beverage popularised on TikTok, whereby half the water in a gallon jug is replaced with alcohol (usually spirits), flavourings, electrolytes and caffeine. We investigated the characteristics and portrayal of the emerging alcohol trend associated with the hashtag descriptor #borg on TikTok. METHODS: We identified highly viewed TikTok videos with the #borg hashtag (n = 103) and conducted a content analysis, capturing viewer engagement ('likes', shares, comments), techniques used, characteristics of featured individuals, and the portrayal of alcohol and risky drinking behaviours. RESULTS: Alcohol was visible in three quarters of the videos analysed (n = 78, average amount of alcohol present 865 mL per borg) and consumed in one third of the videos (n = 34). One quarter of videos (n = 25) promoted alleged benefits of borg consumption compared to other alcohol products or approaches to drinking, yet only nine videos included a warning about potential harms. CONCLUSIONS: The borg trend on TikTok may encourage risky drinking, by portraying it in a style that younger viewers are likely to see as fun and entertaining. SO WHAT?: We were able to gain a better understanding of how this potentially health harming activity is represented on a social media platform that is popular with young people. The speedy dissemination of this trend highlights the need to monitor, investigate and counter emerging trends. Concurrently, there is an urgent need for content restrictions to limit the visibility and promotion of risky alcohol consumption on TikTok.

Workplace interventions to reduce the risk of SARS-CoV-2 infection outside of healthcare settings.

BACKGROUND: Although many people infected with SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) experience no or mild symptoms, some individuals can develop severe illness and may die, particularly older people and those with underlying medical problems. Providing evidence-based interventions to prevent SARS-CoV-2 infection has become more urgent with the potential psychological toll imposed by the coronavirus disease 2019 (COVID-19) pandemic. Controlling exposures to occupational hazards is the fundamental method of protecting workers. When it comes to the transmission of viruses, workplaces should first consider control measures that can potentially have the most significant impact. According to the hierarchy of controls, one should first consider elimination (and substitution), then engineering controls, administrative controls, and lastly, personal protective equipment. This is the first update of a Cochrane review published 6 May 2022, with one new study added. OBJECTIVES: To assess the benefits and harms of interventions in non-healthcare-related workplaces aimed at reducing the risk of SARS-CoV-2 infection compared to other interventions or no intervention. SEARCH METHODS: We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, Embase, Web of Science Core Collections, Cochrane COVID-19 Study Register, World Health Organization (WHO) COVID-19 Global literature on coronavirus disease, ClinicalTrials.gov, the WHO International Clinical Trials Registry Platform, and medRxiv to 13 April 2023. SELECTION CRITERIA: We included randomised controlled trials (RCTs) and non-randomised studies of interventions. We included adult workers, both those who come into close contact with clients or customers (e.g. public-facing employees, such as cashiers or taxi drivers), and those who do not, but who could be infected by coworkers. We excluded studies involving healthcare workers. We included any intervention to prevent or reduce workers' exposure to SARS-CoV-2 in the workplace, defining categories of intervention according to the hierarchy of hazard controls (i.e. elimination; engineering controls; administrative controls; personal protective equipment). DATA COLLECTION AND ANALYSIS: We used standard Cochrane methods. Our primary outcomes were incidence rate of SARS-CoV-2 infection (or other respiratory viruses), SARS-CoV-2-related mortality, adverse events, and absenteeism from work. Our secondary outcomes were all-cause mortality, quality of life, hospitalisation, and uptake, acceptability, or adherence to strategies. We used the Cochrane RoB 2 tool to assess risk of bias, and GRADE methods to evaluate the certainty of evidence for each outcome. MAIN RESULTS: We identified 2 studies including a total of 16,014 participants. Elimination-of-exposure interventions We included one study examining an intervention that focused on elimination of hazards, which was an open-label, cluster-randomised, non-inferiority trial, conducted in England in 2021. The study compared standard 10-day self-isolation after contact with an infected person to a new strategy of daily rapid antigen testing and staying at work if the test is negative (test-based attendance). The trialists hypothesised that this would lead to a similar rate of infections, but lower COVID-related absence. Staff (N = 11,798) working at 76 schools were assigned to standard isolation, and staff (N = 12,229) working at 86 schools were assigned to the test-based attendance strategy. The results between test-based attendance and standard 10-day self-isolation were inconclusive for the rate of symptomatic polymerase chain reaction (PCR)-positive SARS-CoV-2 infection (rate ratio (RR) 1.28, 95% confidence interval (CI) 0.74 to 2.21; 1 study; very low-certainty evidence). The results between test-based attendance and standard 10-day self-isolation were inconclusive for the rate of any PCR-positive SARS-CoV-2 infection (RR 1.35, 95% CI 0.82 to 2.21; 1 study; very low-certainty evidence). COVID-related absenteeism rates were 3704 absence days in 566,502 days-at-risk (6.5 per 1000 working days) in the control group and 2932 per 539,805 days-at-risk (5.4 per 1000 working days) in the intervention group (RR 0.83, 95% CI 0.55 to 1.25). We downgraded the certainty of the evidence to low due to imprecision. Uptake of the intervention was 71% in the intervention group, but not reported for the control intervention. The trial did not measure our other outcomes of SARS-CoV-2-related mortality, adverse events, all-cause mortality, quality of life, or hospitalisation. We found seven ongoing studies using elimination-of-hazard strategies, six RCTs and one non-randomised trial. Administrative control interventions We found one ongoing RCT that aims to evaluate the efficacy of the Bacillus Calmette-Guérin (BCG) vaccine in preventing COVID-19 infection and reducing disease severity. Combinations of eligible interventions We included one non-randomised study examining a combination of elimination of hazards, administrative controls, and personal protective equipment. The study was conducted in two large retail companies in Italy in 2020. The study compared a safety operating protocol, measurement of body temperature and oxygen saturation upon entry, and a SARS-CoV-2 test strategy with a minimum activity protocol. Both groups received protective equipment. All employees working at the companies during the study period were included: 1987 in the intervention company and 1798 in the control company. The study did not report an outcome of interest for this systematic review. Other intervention categories We did not find any studies in this category. AUTHORS' CONCLUSIONS: We are uncertain whether a test-based attendance policy affects rates of PCR-positive SARS-CoV-2 infection (any infection; symptomatic infection) compared to standard 10-day self-isolation amongst school and college staff. A test-based attendance policy may result in little to no difference in absenteeism rates compared to standard 10-day self-isolation. The non-randomised study included in our updated search did not report any outcome of interest for this Cochrane review. As a large part of the population is exposed in the case of a pandemic, an apparently small relative effect that would not be worthwhile from the individual perspective may still affect many people, and thus become an important absolute effect from the enterprise or societal perspective. The included RCT did not report on any of our other primary outcomes (i.e. SARS-CoV-2-related mortality and adverse events). We identified no completed studies on any other interventions specified in this review; however, eight eligible studies are ongoing. More controlled studies are needed on testing and isolation strategies, and working from home, as these have important implications for work organisations.

A simple approach to improving RNA synthesis: Salt inhibition of RNA rebinding coupled with strengthening promoter binding by a targeted gap in the DNA.

T7 RNA polymerase is widely used to synthesize RNA of any length, and long-standing protocols exist to efficiently generate large amounts of RNA. Such synthesis, however, is often plagued by so-called "nontemplated additions" at the 3' end, which are in fact templated by the RNA itself and give rise to double-stranded RNA impurities in RNA therapeutics. These additions are generated by RNA polymerase rebinding to the product RNA (independent of DNA) and this rebinding is in competition with promoter binding. This chapter reports on a general approach that simultaneously weakens RNA rebinding by increasing salt, while at the same time increases promoter binding through manipulating the promoter DNA structure, shifting the balance away from self-primed extension. We present two approaches for use in different regimes. For (short) RNAs using synthetic oligonucleotides as DNA, promoter binding is strengthened by using a partially single stranded promoter construct already in wide use in the field. For the synthesis of RNA (of any length), one can replicate the behavior of the first approach by introducing a targeted gap in the promoter, using a PCR primer containing an engineered deoxyuracil that is then excised by a commercially available enzyme system, to leave a promoter-strengthening gap. Both approaches are simple to implement, with only slight variations on standard synthesis approaches, making them valuable tools for a wide range of applications, from basic science to mRNA, CRISPR, lncRNA, and other therapeutics.

Bis-6-amidino-benzothiazole Derivative that Cures Experimental Stage 1 African Trypanosomiasis with a Single Dose.

We designed and synthesized a series of symmetric bis-6-amidino-benzothiazole derivatives with aliphatic central units and evaluated their efficacy against bloodstream forms of the African trypanosome Trypanosoma brucei. Of these, a dicationic benzothiazole compound (9a) exhibited sub-nanomolar in vitro potency with remarkable selectivity over mammalian cells (>26,000-fold). Unsubstituted 5-amidine groups and a cyclohexyl spacer were the crucial determinants of trypanocidal activity. In all cases, mice treated with a single dose of 20 mg kg-1 were cured of stage 1 trypanosomiasis. The compound displayed a favorable in vitro ADME profile, with the exception of low membrane permeability. However, we found evidence that uptake by T. brucei is mediated by endocytosis, a process that results in lysosomal sequestration. The compound was also active in low nanomolar concentrations against cultured asexual forms of the malaria parasite Plasmodium falciparum. Therefore, 9a has exquisite cross-species efficacy and represents a lead compound with considerable therapeutic potential.

Histories of Medieval Plague in Renaissance Italy.

During the sixteenth century, Italian scholars revised their conception of the field of history so that its purposes went beyond providing political and morally edifying narratives. These scholars contended that history must also account for culture and nature in an encyclopedic fashion. In the same years, numerous newly available texts from antiquity, the Byzantine empire, and the Middle Ages provided insight into the character of earlier outbreaks of plague. Italian physicians, embracing new visions of the field of history, the culture of humanism, and an inductivist epistemology, used these texts to argue that there were continuities among ancient, medieval, and Renaissance epidemics. They catalogued plague and formed historical categories based on severity and perceived origins, leading to the rejection of the conclusions of fourteenth-century western Europeans who viewed the plague of 1347-1353 as unprecedented. These erudite physicians saw medieval plague to be one example of the extreme epidemics that have regularly occurred throughout history.

High-salt transcription from enzymatically gapped promoters nets higher yields and purity of transcribed RNAs.

T7 RNA polymerase is commonly used to synthesize large quantities of RNA for a wide variety of applications, from basic science to mRNA therapeutics. This in vitro system, while showing high fidelity in many ways, is also well known for producing longer than encoded RNA products, particularly under high-yield reaction conditions. Specifically, the resulting product pool is contaminated by an often disperse collection of longer cis-primed extension products. In addition to reducing yield via the conversion of correctly encoded RNA to longer products, self-primed extension generates partially double-stranded RNAs that can trigger the innate immune response. Extensive and low-yield purifications are then required to produce therapeutic RNA. Under high-yield conditions, accumulating concentrations of RNA effectively compete with promoter DNA for polymerase binding, driving self-primed extension at the expense of correct initiation. In the current work, we introduce a simple and novel modification in the DNA to strengthen promoter binding, shifting the balance back toward promoter-driven synthesis and so dramatically reducing self-primed extension. The result is higher yield of the encoded RNA at the outset and reduced need for extensive purifications. The approach can readily be applied to the synthesis of mRNA-length products under high-yield conditions.

The Intersection of Antimicrobial Stewardship, the Pharmaceutical Industry, and the Federal Legislature.

To mitigate the dangers of inappropriate antimicrobial use leading to increased multidrug-resistant organisms and mortality, antimicrobial stewardship programs have become a mainstay in many health systems. Unfortunately, some pharmaceutical manufacturers simultaneously have ended antimicrobial research and development efforts altogether due to suboptimal return on investments. An optimal and sustainable antimicrobial armamentarium requires a broad alliance between antimicrobial stewardship programs, the pharmaceutical industry, the legislature, and federal and state agencies. Public-private relationships such as the Combating Antibiotic-Resistant Bacteria Biopharmaceutical Accelerator (CARB-X) and legislative policies creating push and pull incentives, including the Generating Antibiotic Incentives Now (GAIN), Developing an Innovative Strategy for Antimicrobial-Resistant Microorganisms (DISARM), and Pioneering Antimicrobial Subscriptions to End Upsurging Resistance (PASTEUR) Acts, are each a step in the right direction, but more work remains. Understanding these legislative actions is imperative for all clinicians, as is teamwork from those involved in the antimicrobial field to develop and maintain the life cycle of each drug that harbors societal value.

Workplace interventions to reduce the risk of SARS-CoV-2 infection outside of healthcare settings.

BACKGROUND: Although many people infected with SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2) experience no or mild symptoms, some individuals can develop severe illness and may die, particularly older people and those with underlying medical problems. Providing evidence-based interventions to prevent SARS-CoV-2 infection has become more urgent with the spread of more infectious SARS-CoV-2 variants of concern (VoC), and the potential psychological toll imposed by the coronavirus disease 2019 (COVID-19) pandemic.   Controlling exposures to occupational hazards is the fundamental method of protecting workers. When it comes to the transmission of viruses, such as SARS-CoV-2, workplaces should first consider control measures that can potentially have the most significant impact. According to the hierarchy of controls, one should first consider elimination (and substitution), then engineering controls, administrative controls, and lastly, personal protective equipment (PPE). OBJECTIVES: To assess the benefits and harms of interventions in non-healthcare-related workplaces to reduce the risk of SARS-CoV-2 infection relative to other interventions, or no intervention. SEARCH METHODS: We searched MEDLINE, Embase, Web of Science, Cochrane COVID-19 Study Register, the Canadian Centre for Occupational Health and Safety (CCOHS), Clinicaltrials.gov, and the International Clinical Trials Registry Platform to 14 September 2021. We will conduct an update of this review in six months. SELECTION CRITERIA: We included randomised control trials (RCT) and planned to include non-randomised studies of interventions. We included adult workers, both those who come into close contact with clients or customers (e.g. public-facing employees, such as cashiers or taxi drivers), and those who do not, but who could be infected by co-workers. We excluded studies involving healthcare workers. We included any intervention to prevent or reduce workers' exposure to SARS-CoV-2 in the workplace, defining categories of intervention according to the hierarchy of hazard controls, i.e. elimination; engineering controls; administrative controls; personal protective equipment. DATA COLLECTION AND ANALYSIS: We used standard Cochrane methods. Our primary outcomes were incidence rate of SARS-CoV-2 infection (or other respiratory viruses), SARS-CoV-2-related mortality, adverse events, and absenteeism from work. Our secondary outcomes were all-cause mortality, quality of life, hospitalisation, and uptake, acceptability, or adherence to strategies. We used the Cochrane RoB 2 tool to assess the risk of bias, and GRADE methods to assess the certainty of evidence for each outcome. MAIN RESULTS: Elimination of exposure interventions We included one study examining an intervention that focused on elimination of hazards. This study is an open-label, cluster-randomised, non-inferiority trial, conducted in England in 2021. The study compared standard 10-day self-isolation after contact with an infected person to a new strategy of daily rapid antigen testing and staying at work if the test is negative (test-based attendance). The trialists hypothesised that this would lead to a similar rate of infections, but lower COVID-related absence. Staff (N = 11,798) working at 76 schools were assigned to standard isolation, and staff (N = 12,229) at 86 schools to the test-based attendance strategy.  The results between test-based attendance and standard 10-day self-isolation were inconclusive for the rate of symptomatic PCR-positive SARS-COV-2 infection rate ratio ((RR) 1.28, 95% confidence interval (CI) 0.74 to 2.21; 1 study, very low-certainty evidence)). The results between test-based attendance and standard 10-day self-isolation were inconclusive for the rate of any PCR-positive SARS-COV-2 infection (RR 1.35, 95% CI 0.82 to 2.21; 1 study, very low-certainty evidence). COVID-related absenteeism rates were 3704 absence days in 566,502 days-at-risk (6.5 per 1000 days at risk) in the control group and 2932 per 539,805 days-at-risk (5.4 per 1000 days at risk) in the intervention group (RR 0.83; 95% CI 0.55 to 1.25). The certainty of the evidence was downgraded to low, due to imprecision. Uptake of the intervention was 71 % in the intervention group, but not reported for the control intervention.  The trial did not measure other outcomes, SARS-CoV-2-related mortality, adverse events, all-cause mortality, quality of life, and hospitalisation. We found one ongoing RCT about screening in schools, using elimination of hazard strategies. Personal protective equipment We found one ongoing non-randomised study on the effects of closed face shields to prevent COVID-19 transmission. Other intervention categories We did not find studies in the other intervention categories. AUTHORS' CONCLUSIONS: We are uncertain whether a test-based attendance policy affects rates of PCR-postive SARS-CoV-2 infection (any infection; symptomatic infection) compared to standard 10-day self-isolation amongst school and college staff. Test-based attendance policy may result in little to no difference in absence rates compared to standard 10-day self-isolation. As a large part of the population is exposed in the case of a pandemic, an apparently small relative effect that would not be worthwhile from the individual perspective may still affect many people, and thus, become an important absolute effect from the enterprise or societal perspective.  The included study did not report on any other primary outcomes of our review, i.e. SARS-CoV-2-related mortality and adverse events. No completed studies were identified on any other interventions specified in this review, but two eligible studies are ongoing. More controlled studies are needed on testing and isolation strategies, and working from home, as these have important implications for work organisations.

Medicine and the heavens in Padua's Faculty of Arts, 1570-1630.

In the faculty of arts at the University of Padua in the years around 1600 professors debated the reliability of astrology, the existence of occult celestial influences, and the idea that celestial heat is present in living bodies. From the 1570s to the 1620s many professors in the faculty of arts pushed back against astrology and Jean Fernel's theories surrounding astral body. Girolamo Mercuriale, Alessandro Massaria and Eustachio Rudio thought that some forms of astral causation and Fernel's ideas were incompatible with their observations of disease, Aristotle's philosophy and Hippocratic theories. Later, Santorio Santorio and Cesare Cremonini, who were allied to the political circle of Paolo Sarpi, polemicized against astrology. Their writings show that at Padua medical theory was linked to Aristotelian cosmology, which emphasized the incommensurability between celestial and terrestrial elements. Their rejection of astrology, however, did not lead to the complete marginalization of astrology at Padua. By the middle of the 1620s, as the political climate changed in Venice, the University of Padua hired professors who promoted astrology and Fernel's theories about the celestial nature of innate heat. The diversity of opinions about astrology reflected Venice's divided politics and multiple approaches to interpreting Aristotle and other authorities.

High-salt transcription of DNA cotethered with T7 RNA polymerase to beads generates increased yields of highly pure RNA.

High yields of RNA are routinely prepared following the two-step approach of high-yield in vitro transcription using T7 RNA polymerase followed by extensive purification using gel separation or chromatographic methods. We recently demonstrated that in high-yield transcription reactions, as RNA accumulates in solution, T7 RNA polymerase rebinds and extends the encoded RNA (using the RNA as a template), resulting in a product pool contaminated with longer-than-desired, (partially) double-stranded impurities. Current purification methods often fail to fully eliminate these impurities, which, if present in therapeutics, can stimulate the innate immune response with potentially fatal consequences. In this work, we introduce a novel in vitro transcription method that generates high yields of encoded RNA without double-stranded impurities, reducing the need for further purification. Transcription is carried out at high-salt conditions to eliminate RNA product rebinding, while promoter DNA and T7 RNA polymerase are cotethered in close proximity on magnetic beads to drive promoter binding and transcription initiation, resulting in an increase in overall yield and purity of only the encoded RNA. A more complete elimination of double-stranded RNA during synthesis will not only reduce overall production costs, but also should ultimately enable therapies and technologies that are currently being hampered by those impurities.

Paleocene latitude of the Kohistan-Ladakh arc indicates multistage India-Eurasia collision.

We report paleomagnetic data showing that an intraoceanic Trans-Tethyan subduction zone existed south of the Eurasian continent and north of the Indian subcontinent until at least Paleocene time. This system was active between 66 and 62 Ma at a paleolatitude of 8.1 ± 5.6 °N, placing it 600-2,300 km south of the contemporaneous Eurasian margin. The first ophiolite obductions onto the northern Indian margin also occurred at this time, demonstrating that collision was a multistage process involving at least two subduction systems. Collisional events began with collision of India and the Trans-Tethyan subduction zone in Late Cretaceous to Early Paleocene time, followed by the collision of India (plus Trans-Tethyan ophiolites) with Eurasia in mid-Eocene time. These data constrain the total postcollisional convergence across the India-Eurasia convergent zone to 1,350-2,150 km and limit the north-south extent of northwestern Greater India to <900 km. These results have broad implications for how collisional processes may affect plate reconfigurations, global climate, and biodiversity.

Francis Bacon, José de Acosta, and Traditions of Natural Histories of Winds.

It is well attested that Francis Bacon considered his History of Winds to be an exemplar, but what lessons should be taken from its example have been subject to debate. Instead of looking at this work as a mere model for the fusion of natural history and natural philosophy, it is also possible to see Bacon as trying to provide tentative solutions to outstanding questions regarding the wind, a topic that was deeply scrutinized during the early modern period. An examination of Bacon's provisional concluding rules reveals deep correspondences with earlier works, such as José de Acosta's Natural and Moral History of the Indies, that revised classical understandings of the wind based on experience, experiments, and accounts of travels beyond Europe. Understanding the History of Winds as a genuine attempt to solve outstanding questions about the wind uncovers its debt to earlier traditions, including those related to Renaissance natural history, and shows it's influence in relation to specific theories of the wind beyond its call for methodological reform.

Impact of PhORCAS references on overall application score for postgraduate year 1 pharmacy residency candidates.

PURPOSE: The disparity between the number of applicants for postgraduate year 1 (PGY1) pharmacy residency positions and the number of available residency positions increases the need to optimize how applicants are evaluated. The purpose of the study described here was to evaluate the correlation of ratings of residency candidate characteristics by academic and professional references listed on residency applications with overall application score, applicant ranking, and the likelihood of candidates receiving an invitation to interview. METHODS: A multicenter, retrospective study was conducted to evaluate the correlation of reference writers' ratings of 13 candidate characteristics and their overall recommendations with program-determined outcomes (eg, final application score, applicant ranking, and invitation to interview) through analysis of PGY1 applications submitted through the Pharmacy Online Residency Centralized Application System (PhORCAS) from 2015 through 2018. Keywords and themes within the open-ended section of letters of reference were also analyzed for correlation with overall application score. RESULTS: A total of 5,923 references listed on 1,867 applications to 4 PGY1 pharmacy residency programs processed by PhORCAS were included in the analysis. For the majority of applicant characteristic ratings (ie, 74% of 56,872 ratings overall), reference writers rated candidates as exceeding expectations, and applicants were "highly recommended" by these evaluators in 91% of cases. References' average characteristic ratings and overall recommendations were poorly correlated with final application score (R2 = 0.12 [P < 0.0001] and R2 = 0.08 [P < 0.0001], respectively), final ranking (R2 = 0.02 [P < 0.0001] and R2 = 0.03 [P < 0.0001], respectively), and invitation to interview (R2 = 0.07 [P < 0.0001] and R2 = 0.04 [P < 0.0001], respectively). For the themes evaluated, references' use of teaching words best correlated with normalized final application score, although the correlation was poor (R2 = 0.007, P = 0.0001). CONCLUSION: Reference writers' ratings of PGY1 residency candidate characteristics in PhORCAS are poorly correlated with application score, applicant ranking, and invitation to interview. The results of this study suggest that the existing PhORCAS standardized form for submitting references is of limited utility in its current state.

Sleep quality during an overnight on-call program.

PURPOSE: Many institutions deploy pharmacy residents to expand clinical pharmacy services, often in the form of overnight, in-house on-call programs. There is little published evidence regarding pharmacy resident sleep and sleepiness after a night of overnight, in-house on-call activity. A prospective observational cohort study was conducted to determine the relationship between overnight, in-house on-call programs and pharmacy resident sleep and sleep quality. METHODS: The cohort study included both postgraduate year 1 and postgraduate year 2 pharmacy residents. Each resident participated in 10 to 15 overnight on-call shifts. Sleep and sleep quality were assessed using devices worn on residents' wrists on the nights prior to, during, and after on-call shifts. Resident sleepiness was assessed via the Epworth Sleepiness Scale (ESS) during specified baseline and postcall periods. Univariate and multivariate analysis were used to assess the relationship between measurements of sleep, sleep quality, and sleepiness. RESULTS: We enrolled a total of 23 residents in the study and recorded data on 269 on-call shifts. Frequently (42.6% of shifts) residents had no time to sleep during overnight on-call shifts. Among those who did have sleep time, the mean sleep time during an overnight, in-house on-call shift was 1.22 (SD, 1.56) hours. Additionally, ESS scores indicated a 2.4-fold increase in sleepiness on the morning after vs the morning before on-call shifts. CONCLUSION: Residents often did not sleep while on call. Sleep periods overnight were short and of poor quality. Predictably, residents reported increased sleepiness after an overnight on-call shift. Residents received an average of approximately 10 clinical consultation calls per overnight shift.

Technology-Enabled Person-Centered Mental Health Services Reform: Strategy for Implementation Science.

BACKGROUND: Health information technologies are being rapidly developed to improve the delivery of mental health care; however, a range of facilitators, barriers, and contextual conditions can impact the adoption and sustainment of these solutions. An implementation science protocol supports researchers to achieve primary effectiveness goals in relation to mental health services reform and aids in the optimization of implementation processes to promote quality health care, prolonging sustainability. OBJECTIVE: The aim of this paper is to describe our implementation science protocol, which serves as a foundation by which to systematically guide the implementation of technology-enabled solutions in traditional face-to-face and Web-based mental health services, allowing for revisions over time on the basis of retrospective review and constructive feedback from the services in which the technology-enabled solutions are implemented. METHODS: Our implementation science protocol comprises four phases. The primary objective of the scoping and feasibility phase (Phase 1) is to determine the alignment between the service partner and the quality improvement goals supported by the technology-enabled solution. This is followed by Phase 2, the local co-design and preimplementation phase, which aims to utilize co-design methodologies, including service pathway modelling, participatory design, and user (acceptance) testing, to determine how the solutions could be used to enhance the service. In Phase 3, implementation, the accepted solution is embedded in the mental health service to achieve better outcomes for consumers and their families as well as health professionals and service managers. Using iterative evaluative processes throughout Phase 3, the solution is continuously developed, designed, and refined during implementation to adapt to the changing needs of the stakeholders, including consumers with lived experience and their families as well as the service. Thus, the primary outcome of Phase 3 is the optimized technology-enabled solution that can be maintained in a service during the sustainment and scalability phase (Phase 4) for the purposes of mental health services reform. RESULTS: Funding for the protocol was provided by the Australian Government Department of Health in June of 2017 for a period of 3 years. At the time of this publication, the protocol had been initiated in 11 services, serving three populations, all of which are currently operating in Phase 3. The first results are expected to be submitted for publication in 2020. CONCLUSIONS: With the aim of improving mental health service quality, our implementation science protocol aids in the identification of factors that predict the likelihood of implementation success, as well as the development of strategies to proactively mitigate potential barriers to achieve better implementation outcomes. Putting in place a theoretically sound implementation science protocol is essential to facilitate the uptake of novel technology-enabled solutions and evidence-based practices into routine clinical practice for the purposes of improved outcomes.

Efficient inhibition of RNA self-primed extension by addition of competing 3'-capture DNA-improved RNA synthesis by T7 RNA polymerase.

In vitro synthesized RNA is used widely in studies of RNA biology, biotechnology and RNA therapeutics. However, in vitro synthesized RNA often contains impurities, such as RNAs with lengths shorter and longer than the expected runoff RNA. We have recently confirmed that longer RNA products are formed predominantly via cis self-primed extension, in which released runoff RNA folds back on itself to prime its own RNA-templated extension. In the current work, we demonstrate that addition of a DNA oligonucleotide (capture DNA) that is complementary to the 3' end of the expected runoff RNA effectively prevents self-primed extension, even under conditions commonly used for high RNA yields. Moreover, the presence of this competing capture DNA during 'high yield' transcription, leads to an increase in the yield of expected runoff RNA by suppressing the formation of undesired longer RNA byproducts.

External water transport is more important than vascular transport in the extreme atmospheric epiphyte Tillandsia usneoides (Spanish moss).

Most epiphytic bromeliads, especially those in the genus Tillandsia, lack functional roots and rely on the absorption of water and nutrients by large, multicellular trichomes on the epidermal surfaces of leaves and stems. Another important function of these structures is the spread of water over the epidermal surface by capillary action between trichome "wings" and epidermal surface. Although critical for the ultimate absorption by these plants, understanding of this function of trichomes is primarily based on light microscope observations. To better understand this phenomenon, the distribution of water was followed by its attenuation of cold neutrons following application of H2 O to the cut end of Tillandsia usneoides shoots. Experiments confirmed the spread of added water on the external surfaces of this "atmospheric" epiphyte. In a morphologically and physiologically similar plant lacking epidermal trichomes, water added to the cut end of a shoot clearly moved via its internal xylem and not on its epidermis. Thus, in T. usneoides, water moves primarily by capillarity among the overlapping trichomes forming a dense indumentum on shoot surfaces, while internal vascular water movement is less likely. T. usneoides, occupying xeric microhabitats, benefits from reduction of water losses by low-shoot xylem hydraulic conductivities.

3' end additions by T7 RNA polymerase are RNA self-templated, distributive and diverse in character-RNA-Seq analyses.

Synthetic RNA is widely used in basic science, nanotechnology and therapeutics research. The vast majority of this RNA is synthesized in vitro by T7 RNA polymerase or one of its close family members. However, the desired RNA is generally contaminated with products longer and shorter than the DNA-encoded product. To better understand these undesired byproducts and the processes that generate them, we analyze in vitro transcription reactions using RNA-Seq as a tool. The results unambiguously confirm that product RNA rebinds to the polymerase and self-primes (in cis) generation of a hairpin duplex, a process that favorably competes with promoter driven synthesis under high yield reaction conditions. While certain priming modes can be favored, the process is heterogeneous, both in initial priming and in the extent of priming, and already extended products can rebind for further extension, in a distributive process. Furthermore, addition of one or a few nucleotides, previously termed 'nontemplated addition,' also occurs via templated primer extension. At last, this work demonstrates the utility of RNA-Seq as a tool for in vitro mechanistic studies, providing information far beyond that provided by traditional gel electrophoresis.

Expanding the toolbox for Trypanosoma cruzi: A parasite line incorporating a bioluminescence-fluorescence dual reporter and streamlined CRISPR/Cas9 functionality for rapid in vivo localisation and phenotyping.

BACKGROUND: Infection with Trypanosoma cruzi causes Chagas disease, a major public health problem throughout Latin America. There is no vaccine and the only drugs have severe side effects. Efforts to generate new therapies are hampered by limitations in our understanding of parasite biology and disease pathogenesis. Studies are compromised by the complexity of the disease, the long-term nature of the infection, and the fact that parasites are barely detectable during the chronic stage. In addition, functional dissection of T. cruzi biology has been restricted by the limited flexibility of the genetic manipulation technology applicable to this parasite. METHODOLOGY/PRINCIPAL FINDINGS: Here, we describe two technical innovations, which will allow the role of the parasite in disease progression to be better assessed. First, we generated a T. cruzi reporter strain that expresses a fusion protein comprising red-shifted luciferase and green fluorescent protein domains. Bioluminescence allows the kinetics of infection to be followed within a single animal, and specific foci of infection to be pinpointed in excised tissues. Fluorescence can then be used to visualise individual parasites in tissue sections to study host-parasite interactions at a cellular level. Using this strategy, we have been routinely able to find individual parasites within chronically infected murine tissues for the first time. The second advance is the incorporation of a streamlined CRISPR/Cas9 functionality into this reporter strain that can facilitate genome editing using a PCR-based approach that does not require DNA cloning. This system allows the rapid generation of null mutants and fluorescently tagged parasites in a background where the in vivo phenotype can be rapidly assessed. CONCLUSIONS/SIGNIFICANCE: The techniques described here will have multiple applications for studying aspects of T. cruzi biology and Chagas disease pathogenesis previously inaccessible to conventional approaches. The reagents and cell lines have been generated as a community resource and are freely available on request.