High-throughput crystallographic fragment screening of Zika virus NS3 Helicase.

The Zika virus (ZIKV), discovered in Africa in 1947, swiftly spread across continents, causing significant concern due to its recent association with microcephaly in newborns and Guillain-Barré syndrome in adults. Despite a decrease in prevalence, the potential for a resurgence remains, necessitating urgent therapeutic interventions. Like other flaviviruses, ZIKV presents promising drug targets within its replication machinery, notably the NS3 helicase (NS3 Hel ) protein, which plays critical roles in viral replication. However, a lack of structural information impedes the development of specific inhibitors targeting NS3 Hel . Here we applied high-throughput crystallographic fragment screening on ZIKV NS3 Hel , which yielded structures that reveal 3D binding poses of 46 fragments at multiple sites of the protein, including 11 unique fragments in the RNA-cleft site. These fragment structures provide templates for direct design of hit compounds and should thus assist the development of novel direct-acting antivirals against ZIKV and related flaviviruses, thus opening a promising avenue for combating future outbreaks.

Crystallographic fragment screening delivers diverse chemical scaffolds for Zika virus NS2B-NS3 protease inhibitor development.

Zika virus (ZIKV) infections cause microcephaly in new-borns and Guillain-Barre syndrome in adults raising a significant global public health concern, yet no vaccines or antiviral drugs have been developed to prevent or treat ZIKV infections. The viral protease NS3 and its co-factor NS2B are essential for the cleavage of the Zika polyprotein precursor into individual structural and non-structural proteins and is therefore an attractive drug target. Generation of a robust crystal system of co-expressed NS2B-NS3 protease has enabled us to perform a crystallographic fragment screening campaign with 1076 fragments. 48 binders with diverse chemical scaffolds were identified in the active site of the protease, with another 6 fragment hits observed in a potential allosteric binding site. Our work provides potential starting points for the development of potent NS2B-NS3 protease inhibitors. Furthermore, we have structurally characterized a potential allosteric binding pocket, identifying opportunities for allosteric inhibitor development.

Crystallographic Fragment Screen of Coxsackievirus A16 2A Protease identifies new opportunities for the development of broad-spectrum anti-enterovirals.

Enteroviruses are the causative agents of paediatric hand-foot-and-mouth disease, and a target for pandemic preparedness due to the risk of higher order complications in a large-scale outbreak. The 2A protease of these viruses is responsible for the self-cleavage of the poly protein, allowing for correct folding and assembly of capsid proteins in the final stages of viral replication. These 2A proteases are highly conserved between Enterovirus species, such as Enterovirus A71 and Coxsackievirus A16 . Inhibition of the 2A protease deranges capsid folding and assembly, preventing formation of mature virions in host cells and making the protease a valuable target for antiviral activity. Herein, we describe a crystallographic fragment screening campaign that identified 75 fragments which bind to the 2A protease including 38 unique compounds shown to bind within the active site. These fragments reveal a path for the development of non-peptidomimetic inhibitors of the 2A protease with broad-spectrum anti-enteroviral activity.

Highly selective CO2 photoreduction to CO on MOF-derived TiO2.

Metal-Organic Framework (MOF)-derived TiO2, synthesised through the calcination of MIL-125-NH2, is investigated for its potential as a CO2 photoreduction catalyst. The effect of the reaction parameters: irradiance, temperature and partial pressure of water was investigated. Using a two-level design of experiments, we were able to evaluate the influence of each parameter and their potential interactions on the reaction products, specifically the production of CO and CH4. It was found that, for the explored range, the only statistically significant parameter is temperature, with an increase in temperature being correlated to enhanced production of both CO and CH4. Over the range of experimental settings explored, the MOF-derived TiO2 displays high selectivity towards CO (98%), with only a small amount of CH4 (2%) being produced. This is notable when compared to other state-of-the-art TiO2 based CO2 photoreduction catalysts, which often showcase lower selectivity. The MOF-derived TiO2 was found to have a peak production rate of 8.9 × 10-4 µmol cm-2 h-1 (2.6 µmol g-1 h-1) and 2.6 × 10-5 µmol cm-2 h-1 (0.10 µmol g-1 h-1) for CO and CH4, respectively. A comparison is made to commercial TiO2, P25 (Degussa), which was shown to have a similar activity towards CO production, 3.4 × 10-3 µmol cm-2 h-1 (5.9 µmol g-1 h-1), but a lower selectivity preference for CO (3 : 1 CH4 : CO) than the MOF-derived TiO2 material developed here. This paper showcases the potential for MIL-125-NH2 derived TiO2 to be further developed as a highly selective CO2 photoreduction catalyst for CO production.

Enhancing behavioral intervention science: using community-based participatory research principles with the multiphase optimization strategy.

Innovative methodological frameworks are needed in intervention science to increase efficiency, potency, and community adoption of behavioral health interventions, as it currently takes 17 years and millions of dollars to test and disseminate interventions. The multiphase optimization strategy (MOST) for developing behavioral interventions was designed to optimize efficiency, efficacy, and sustainability, while community-based participatory research (CBPR) engages community members in all research steps. Classical approaches for developing behavioral interventions include testing against control interventions in randomized controlled trials. MOST adds an optimization phase to assess performance of individual intervention components and their interactions on outcomes. This information is used to engineer interventions that meet specific optimization criteria focused on effectiveness, cost, or time. Combining CBPR and MOST facilitates development of behavioral interventions that effectively address complex health challenges, are acceptable to communities, and sustainable by maximizing resources, building community capacity and acceptance. Herein, we present a case study to illustrate the value of combining MOST and CBPR to optimize a multilevel intervention for reducing substance misuse among formerly incarcerated men, for under $250 per person. This integration merged experiential and cutting-edge scientific knowledge and methods, built community capacity, and promoted the development of efficient interventions. Integrating CBPR and MOST principles yielded a framework of intervention development/testing that is more efficient, faster, cheaper, and rigorous than traditional stage models. Combining MOST and CBPR addressed significant intervention science gaps and speeds up testing and implementation of interventions.

Synthesis of TiO2-x/W18O49 hollow double-shell and core-shell microspheres for CO2 photoreduction under visible light.

TiO2-x/W18O49 with core-shell or double-shelled hollow microspheres were synthesized through a facile multi-step solvothermal method. The formation of the hollow microspheres with a double-shell was a result of the Kirkendall effect during the solvothermal treatment with concentrated NaOH. The advanced architecture significantly enhanced the electronic properties of TiO2-x/W18O49, improving by more than 30 times the CO2 photoreduction efficiency compared to the pristine W18O49. Operando DRIFTS measurements revealed that the yellow TiO2-x was a preferable CO2 adsorption and conversion site.

Alkali modified P25 with enhanced CO2 adsorption for CO2 photoreduction.

To improve the CO2 adsorption on the photocatalyst, which is an essential step for CO2 photoreduction, solid solutions were fabricated using a facile calcination treatment at 900 °C. Using various alkalis, namely NaOH, Na2CO3, KOH, K2CO3, the resulted samples presented a much higher CO2 adsorption capacity, which was measured with the pulse injection of CO2 on the temperature programmed desorption workstation, compared to the pristine Evonik P25. As a result, all of the fabricated solid solutions produced higer yield of CO under UV light irradiation due to the increased basicity of the solid solutions even though they possessed only the rutile polymorph of TiO2. The highest CO2 adsorption capacity under UV irradiation was observed in the sample treated with NaOH, which contained the highest amount of isolated hydroxyls, as shown in the FTIR studies.

Systematic study of TiO2/ZnO mixed metal oxides for CO2 photoreduction.

A two component three degree simplex lattice experimental design was employed to evaluate the impact of different mixing fractions of TiO2 and ZnO on an ordered mesoporous SBA-15 support for CO2 photoreduction. It was anticipated that the combined advantages of TiO2 and ZnO: low cost, non-toxicity and combined electronic properties would facilitate CO2 photoreduction. The fraction of ZnO had a statistically dominant impact on maximum CO2 adsorption (ß 2 = 22.65, p-value = 1.39 × 10-4). The fraction of TiO2 used had a statistically significant positive impact on CO (ß 1 = 9.71, p-value = 2.93 × 10-4) and CH4 (ß 1 = 1.43, p-value = 1.35 × 10-3) cumulative production. A negative impact, from the interaction term between the fractions of TiO2 and ZnO, was found for CH4 cumulative production (ß 3 = -2.64, p-value = 2.30 × 10-2). The systematic study provided evidence for the possible loss in CO2 photoreduction activity from sulphate groups introduced during the synthesis of ZnO. The decrease in activity is attributed to the presence of sulphate species in the ZnO prepared, which may possibly act as charge carrier and/or radical intermediate scavengers.

The effect of culinary interventions (cooking classes) on dietary intake and behavioral change: a systematic review and evidence map.

BACKGROUND: Culinary interventions (cooking classes) have been used to improve the quality of dietary intake and change behavior. The aim of this systematic review is to investigate the effects of culinary interventions on dietary intake and behavioral and cardiometabolic outcomes. METHODS: We conducted a systematic review of MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews, and Scopus for comparative studies that evaluated culinary interventions to a control group or baseline values. The intervention was defined as a cooking class regardless of its length or delivery approach. Studies included populations of children, healthy adults or adults with morbidities. The risk of bias was assessed using the Cochrane Risk of Bias tool and the Newcastle-Ottawa Scale. Outcomes were pooled using the random-effects model and descriptive statistics and depicted in an evidence map. Simple logistic regression was used to evaluate factors associated with intervention success. RESULTS: We included 30 studies (6 were randomized, 7381 patients, average follow up 25 weeks). Culinary interventions were not associated with a significant change in body mass index (- 0.07 kg/m2, 95% CI: -1.53, 1.40), systolic (- 5.31 mmHg, 95% CI: -34.2, 23.58) or diastolic blood pressure (- 3.1 mmHg, 95% CI: -23.82, 17.62) or LDL cholesterol (- 8.09 mg/dL, 95% CI: -84.43, 68.25). Culinary interventions were associated with improved attitudes, self-efficacy and healthy dietary intake in adults and children. We were unable to demonstrate whether the effect of a culinary intervention was modified by various characteristics of the intervention such as its delivery or intensity. Interventions with additional components such as education on nutrition, physical activity or gardening were particularly effective. CONCLUSIONS: Culinary interventions were not associated with a significant change in cardiometabolic risk factors, but were associated with improved attitudes, self-efficacy and a healthier dietary intake in adults and children.

Occupation, Sitting, and Weight Change in a Cohort of Women Employees.

OBJECTIVE: Few studies have looked at occupation and weight gain over time. We examined the influence of occupation on sitting and weight change in employed women. METHODS: A total of 228 women working as appointment coordinators or clinical assistants were surveyed regarding sitting and physical activity. Medical records were reviewed to determine changes in weight while employed in that position. Follow-up averaged 6.9 years. RESULTS: Eight hours or more of sitting daily was seen in 74% of appointment coordinators and 38% of clinical assistants (P < 0.001). Appointment coordinators were not as physically active (P = 0.026) and gained more weight (P = 0.045) over time than clinical assistants. Controlling for physical activity modestly attenuated the effect of occupation on weight gain over time (P = 0.061). CONCLUSIONS: Occupation has a profound influence on sitting and may influence physical activity and weight gain over time.

Enhancing motivation with the "virtual" supervisory role: a randomized trial.

BACKGROUND: We aimed to explore the influence of a motivationally-enhanced instructional design on motivation to learn and knowledge, hypothesizing that outcomes would be higher for the enhanced instructional format. METHODS: Medicine residents completed four online learning modules on primary care topics. Using a crossover design, learners were randomized to receive two standard and two motivationally-enhanced learning modules. Both formats had self-assessment questions, but the enhanced format questions were framed to place learners in a supervisory/teaching role. Learners received a baseline motivation questionnaire, a short motivation survey before and after each module, and a knowledge posttest. RESULTS: One hundred twenty seven residents were randomized. 123 residents (97%) completed at least one knowledge posttest and 119 (94%) completed all four posttests. Across all modules, a one-point increase in the pretest short motivation survey was associated with a 2.1-point increase in posttest knowledge. The change in motivation was significantly higher for the motivationally enhanced format (standard mean change -0.01, enhanced mean change +0.09, difference = 0.10, CI 0.001 to 0.19; p = 0.048). Mean posttest knowledge score was similar (standard mean 72.8, enhanced mean 73.0, difference = 0.2, CI -1.9 to 2.1; p = 0.90). CONCLUSIONS: The motivationally enhanced instructional format improved motivation more than the standard format, but impact on knowledge scores was small and not statistically significant. Learners with higher pre-intervention motivation scored better on post-intervention knowledge tests, suggesting that motivation may prove a viable target for future instructional enhancements.

A summary and critical assessment of the 2013 ACC/AHA guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular disease risk in adults: filling the gaps.

The American College of Cardiology/American Heart Association (ACC/AHA) Task Force on Practice Guidelines has recently released the new cholesterol treatment guideline. This update was based on a systematic review of the evidence and replaces the previous guidelines from 2002 that were widely accepted and implemented in clinical practice. The new cholesterol treatment guideline emphasizes matching the intensity of statin treatment to the level of atherosclerotic cardiovascular disease (ASCVD) risk and replaces the old paradigm of pursuing low-density lipoprotein cholesterol targets. The new guideline also emphasizes the primacy of the evidence base for statin therapy for ASCVD risk reduction and lists several patient groups that will not benefit from statin treatment despite their high cardiovascular risk, such as those with heart failure (New York Heart Association class II-IV) and patients undergoing hemodialysis. The guideline has been received with mixed reviews and significant controversy. Because of the evidence-based nature of the guideline, there is room for several questions and uncertainties on when and how to use lipid-lowering therapy in clinical practice. The goal of the Mayo Clinic Task Force in the assessment, interpretation, and expansion of the ACC/AHA cholesterol treatment guideline is to address gaps in information and some of the controversial aspects of the newly released cholesterol management guideline using additional sources of evidence and expert opinion as needed to guide clinicians on key aspects of ASCVD risk reduction.

Comfort and experience with online learning: trends over nine years and associations with knowledge.

BACKGROUND: Some evidence suggests that attitude toward computer-based instruction is an important determinant of success in online learning. We sought to determine how comfort using computers and perceptions of prior online learning experiences have changed over the past decade, and how these associate with learning outcomes. METHODS: Each year from 2003-2011 we conducted a prospective trial of online learning. As part of each year's study, we asked medicine residents about their comfort using computers and if their previous experiences with online learning were favorable. We assessed knowledge using a multiple-choice test. We used regression to analyze associations and changes over time. RESULTS: 371 internal medicine and family medicine residents participated. Neither comfort with computers nor perceptions of prior online learning experiences showed a significant change across years (p > 0.61), with mean comfort rating 3.96 (maximum 5 = very comfortable) and mean experience rating 4.42 (maximum 6 = strongly agree [favorable]). Comfort showed no significant association with knowledge scores (p = 0.39) but perceptions of prior experiences did, with a 1.56% rise in knowledge score for a 1-point rise in experience score (p = 0.02). Correlations among comfort, perceptions of prior experiences, and number of prior experiences were all small and not statistically significant. CONCLUSIONS: Comfort with computers and perceptions of prior experience with online learning remained stable over nine years. Prior good experiences (but not comfort with computers) demonstrated a modest association with knowledge outcomes, suggesting that prior course satisfaction may influence subsequent learning.

"Go4Life" exercise counseling, accelerometer feedback, and activity levels in older people.

Older people are more sedentary than other age groups. We sought to determine if providing an accelerometer with feedback about activity and counseling older subjects using Go4Life educational material would increase activity levels. Participants were recruited from independent living areas within assisted living facilities and the general public in the Rochester, MN area. 49 persons aged 65-95 (79.5±7.0 years) who were ambulatory but sedentary and overweight participated in this randomized controlled crossover trial for one year. After a baseline period of 2 weeks, group 1 received an accelerometer and counseling using Go4Life educational material (www.Go4Life.nia.nih.gov) for 24 weeks and accelerometer alone for the next 24 weeks. Group 2 had no intervention for the first 24 weeks and then received an accelerometer and Go4Life based counseling for 24 weeks. There were no significant baseline differences between the two groups. The intervention was not associated with a significant change in activity, body weight, % body fat, or blood parameters (p>0.05). Older (80-93) subjects were less active than younger (65-79) subjects (p=0.003). Over the course of the 48 week study, an increase in activity level was associated with a decline in % body fat (p=0.008). Increasing activity levels benefits older patients. However, providing an accelerometer and a Go4Life based exercise counseling program did not result in a 15% improvement in activity levels in this elderly population. Alternate approaches to exercise counseling may be needed in elderly people of this age range.

Test-enhanced web-based learning: optimizing the number of questions (a randomized crossover trial).

PURPOSE: Questions enhance learning in Web-based courses, but preliminary evidence suggests that too many questions may interfere with learning. The authors sought to determine how varying the number of self-assessment questions affects knowledge outcomes in a Web-based course. METHOD: The authors conducted a randomized crossover trial in one internal medicine and one family medicine residency program between January 2009 and July 2010. Eight Web-based modules on ambulatory medicine topics were developed, with varying numbers of self-assessment questions (0, 1, 5, 10, or 15). Participants completed modules in four different formats each year, with sequence randomly assigned. Participants completed a pretest for half their modules. Outcomes included knowledge, completion time, and module ratings. RESULTS: One hundred eighty residents provided data. The mean (standard error) percent correct knowledge score was 53.2 (0.8) for pretests and 73.7 (0.5) for posttests. In repeated-measures analysis pooling all data, mean posttest knowledge scores were highest for the 10- and 15-question formats (75.7 [1.1] and 74.4 [1.0], respectively) and lower for 0-, 1-, and 5-question formats (73.1 [1.3], 72.9 [1.0], and 72.8 [1.5], respectively); P = .04 for differences across all modules. Modules with more questions generally took longer to complete and were rated higher, although differences were small. Residents most often identified 10 questions as ideal. Posttest knowledge scores were higher for modules that included a pretest (75.4 [0.9] versus 72.2 [0.9]; P = .0002). CONCLUSIONS: Increasing the number of self-assessment questions improves learning until a plateau beyond which additional questions do not add value.

Increasing physician activity with treadmill desks.

BACKGROUND: Prolonged sitting has been shown to increase mortality and obesity. OBJECTIVE: We sought to determine whether physicians would use a treadmill desk, increase their daily physical activity and lose weight. PARTICIPANTS: 20 overweight and obese physicians aged 25 to 70 with Body Mass Index > 25. METHODS: Participants used a treadmill desk, a triaxial accelerometer, and received exercise counseling in a randomized, cross-over trial over 24 weeks. Group 1 received exercise counseling, accelerometer feedback, and a treadmill desk for 12 weeks and then accelerometer only for 12 weeks. Group 2 received an accelerometer without feedback for 12 weeks followed by exercise counseling, accelerometer feedback, and the treadmill desk for 12 weeks. RESULTS: Daily physical activity increased while using the treadmill desk compared to not using the desk by 197 kcal per day (p=0.003). The difference in weight during the two 12 week periods was 1.85 kg (p=0.03). Percent body fat was 1.9% lower while using the treadmill desk (p=0.02). There were no differences in metabolic or well-being measures. CONCLUSIONS: This study suggests that physicians will use a treadmill desk, that it does increase their activity, and that it may help with weight loss. Further studies are warranted.

Absorptions between 3000 and 5500 cm(-1) of cyclic O4+ and O4- trapped in solid neon.

Recently, gas-phase absorptions in the 3000-4300 cm(-1) spectral region have been assigned to combination bands built on (ν1 + ν5) of ground-state cyc-O4(+). Other gas-phase experiments identified an electronic transition of cyc-O4(-) complexed with an argon atom between 4000 and 5300 cm(-1). Absorptions that correspond closely to these two groups of bands have been observed in neon-matrix experiments in which both cyc-O4(+) and cyc-O4(-) are trapped at 4.3 K in solid neon. The results are compared with the gas-phase data, and the proposed assignments are considered by taking into account the results of isotopic substitution.