Modelling and Molecular Dynamics Predict the Structure and Interactions of the Glycine Receptor Intracellular Domain.

Glycine receptors (GlyRs) are glycine-gated inhibitory pentameric ligand-gated ion channels composed of α or α + ß subunits. A number of structures of these proteins have been reported, but to date, these have only revealed details of the extracellular and transmembrane domains, with the intracellular domain (ICD) remaining uncharacterised due to its high flexibility. The ICD is a region that can modulate function in addition to being critical for receptor localisation and clustering via proteins such as gephyrin. Here, we use modelling and molecular dynamics (MD) to reveal details of the ICDs of both homomeric and heteromeric GlyR. At their N and C ends, both the α and ß subunit ICDs have short helices, which are major sites of stabilising interactions; there is a large flexible loop between them capable of forming transient secondary structures. The α subunit can affect the ß subunit ICD structure, which is more flexible in a 4α2:1ß than in a 4α1:1ß GlyR. We also explore the effects of gephyrin binding by creating GlyR models bound to the gephyrin E domain; MD simulations suggest these are more stable than the unbound forms, and again there are α subunit-dependent differences, despite the fact the gephyrin binds to the ß subunit. The bound models also suggest that gephyrin causes compaction of the ICD. Overall, the data expand our knowledge of this important receptor protein and in particular clarify features of the underexplored ICD.

SARS-CoV-2 Omicron subvariant spike N405 unlikely to rapidly deamidate.

The RGD motif on the SARS-CoV-2 spike protein has been suggested to interact with RGD-binding integrins αVß3 and α5ß1 to enhance viral cell entry and alter downstream signaling cascades. The D405N mutation on the Omicron subvariant spike proteins, resulting in an RGN motif, has recently been shown to inhibit binding to integrin αVß3. Deamidation of asparagines in protein ligand RGN motifs has been demonstrated to generate RGD and RGisoD motifs that permit binding to RGD-binding integrins. Two asparagines, N481 and N501, on the Wild-type spike receptor-binding domain have been previously shown to have deamidation half-lives of 16.5 and 123 days, respectively, which may occur during the viral life cycle. Deamidation of Omicron subvariant N405 may recover the ability to interact with RGD-binding integrins. Thus, herein, all-atom molecular dynamics simulations of the Wild-type and Omicron subvariant spike protein receptor-binding domains were conducted to investigate the potential for asparagines, the Omicron subvariant N405 in particular, to assume the optimized geometry for deamidation to occur. In summary, the Omicron subvariant N405 was primarily found to be stabilized in a state unfavourable for deamidation after hydrogen bonding with downstream E406. Nevertheless, a small number of RGD or RGisoD motifs on the Omicron subvariant spike proteins may restore the ability to interact with RGD-binding integrins. The simulations also provided structural clarification regarding the deamidation rates of Wild-type N481 and N501 and highlighted the utility of tertiary structure dynamics information in predicting asparagine deamidation. Further work is needed to characterize the effects of deamidation on spike-integrin interactions.

In silico analysis of mutations near S1/S2 cleavage site in SARS-CoV-2 spike protein reveals increased propensity of glycosylation in Omicron strain.

Cleavage of the severe respiratory syndrome coronavirus-2 (SARS-CoV-2) spike protein has been demonstrated to contribute to viral-cell fusion and syncytia formation. Studies have shown that variants of concern (VOC) and variants of interest (VOI) show differing membrane fusion capacity. Mutations near cleavage motifs, such as the S1/S2 and S2' sites, may alter interactions with host proteases and, thus, the potential for fusion. The biochemical basis for the differences in interactions with host proteases for the VOC/VOI spike proteins has not yet been explored. Using sequence and structure-based bioinformatics, mutations near the VOC/VOI spike protein cleavage sites were inspected for their structural effects. All mutations found at the S1/S2 sites were predicted to increase affinity to the furin protease but not TMPRSS2. Mutations at the spike residue P681 in several strains, such P681R in the Delta strain, resulted in the disruption of a proline-directed kinase phosphorylation motif at the S1/S2 site, which may lessen the impact of phosphorylation for these variants. However, the unique N679K mutation in the Omicron strain was found to increase the propensity for O-linked glycosylation at the S1/S2 cleavage site, which may prevent recognition by proteases. Such glycosylation in the Omicron strain may hinder entry at the cell surface and, thus, decrease syncytia formation and induce cell entry through the endocytic pathway as has been shown in previous studies. Further experimental work is needed to confirm the effect of mutations and posttranslational modifications on SARS-CoV-2 spike protein cleavage sites.

Unheeded SARS-CoV-2 proteins? A deep look into negative-sense RNA.

SARS-CoV-2 is a novel positive-sense single-stranded RNA virus from the Coronaviridae family (genus Betacoronavirus), which has been established as causing the COVID-19 pandemic. The genome of SARS-CoV-2 is one of the largest among known RNA viruses, comprising of at least 26 known protein-coding loci. Studies thus far have outlined the coding capacity of the positive-sense strand of the SARS-CoV-2 genome, which can be used directly for protein translation. However, it has been recently shown that transcribed negative-sense viral RNA intermediates that arise during viral genome replication from positive-sense viruses can also code for proteins. No studies have yet explored the potential for negative-sense SARS-CoV-2 RNA intermediates to contain protein-coding loci. Thus, using sequence and structure-based bioinformatics methodologies, we have investigated the presence and validity of putative negative-sense ORFs (nsORFs) in the SARS-CoV-2 genome. Nine nsORFs were discovered to contain strong eukaryotic translation initiation signals and high codon adaptability scores, and several of the nsORFs were predicted to interact with RNA-binding proteins. Evolutionary conservation analyses indicated that some of the nsORFs are deeply conserved among related coronaviruses. Three-dimensional protein modeling revealed the presence of higher order folding among all putative SARS-CoV-2 nsORFs, and subsequent structural mimicry analyses suggest similarity of the nsORFs to DNA/RNA-binding proteins and proteins involved in immune signaling pathways. Altogether, these results suggest the potential existence of still undescribed SARS-CoV-2 proteins, which may play an important role in the viral lifecycle and COVID-19 pathogenesis.

Are There Hidden Genes in DNA/RNA Vaccines?

Due to the fast global spreading of the Severe Acute Respiratory Syndrome Coronavirus - 2 (SARS-CoV-2), prevention and treatment options are direly needed in order to control infection-related morbidity, mortality, and economic losses. Although drug and inactivated and attenuated virus vaccine development can require significant amounts of time and resources, DNA and RNA vaccines offer a quick, simple, and cheap treatment alternative, even when produced on a large scale. The spike protein, which has been shown as the most antigenic SARS-CoV-2 protein, has been widely selected as the target of choice for DNA/RNA vaccines. Vaccination campaigns have reported high vaccination rates and protection, but numerous unintended effects, ranging from muscle pain to death, have led to concerns about the safety of RNA/DNA vaccines. In parallel to these studies, several open reading frames (ORFs) have been found to be overlapping SARS-CoV-2 accessory genes, two of which, ORF2b and ORF-Sh, overlap the spike protein sequence. Thus, the presence of these, and potentially other ORFs on SARS-CoV-2 DNA/RNA vaccines, could lead to the translation of undesired proteins during vaccination. Herein, we discuss the translation of overlapping genes in connection with DNA/RNA vaccines. Two mRNA vaccine spike protein sequences, which have been made publicly-available, were compared to the wild-type sequence in order to uncover possible differences in putative overlapping ORFs. Notably, the Moderna mRNA-1273 vaccine sequence is predicted to contain no frameshifted ORFs on the positive sense strand, which highlights the utility of codon optimization in DNA/RNA vaccine design to remove undesired overlapping ORFs. Since little information is available on ORF2b or ORF-Sh, we use structural bioinformatics techniques to investigate the structure-function relationship of these proteins. The presence of putative ORFs on DNA/RNA vaccine candidates implies that overlapping genes may contribute to the translation of smaller peptides, potentially leading to unintended clinical outcomes, and that the protein-coding potential of DNA/RNA vaccines should be rigorously examined prior to administration.

Searching for New Z-DNA/Z-RNA Binding Proteins Based on Structural Similarity to Experimentally Validated Zα Domain.

Z-DNA and Z-RNA are functionally important left-handed structures of nucleic acids, which play a significant role in several molecular and biological processes including DNA replication, gene expression regulation and viral nucleic acid sensing. Most proteins that have been proven to interact with Z-DNA/Z-RNA contain the so-called Zα domain, which is structurally well conserved. To date, only eight proteins with Zα domain have been described within a few organisms (including human, mouse, Danio rerio, Trypanosoma brucei and some viruses). Therefore, this paper aimed to search for new Z-DNA/Z-RNA binding proteins in the complete PDB structures database and from the AlphaFold2 protein models. A structure-based similarity search found 14 proteins with highly similar Zα domain structure in experimentally-defined proteins and 185 proteins with a putative Zα domain using the AlphaFold2 models. Structure-based alignment and molecular docking confirmed high functional conservation of amino acids involved in Z-DNA/Z-RNA, suggesting that Z-DNA/Z-RNA recognition may play an important role in a variety of cellular processes.

Can the SARS-CoV-2 Spike Protein Bind Integrins Independent of the RGD Sequence?

The RGD motif in the Severe Acute Syndrome Coronavirus 2 (SARS-CoV-2) spike protein has been predicted to bind RGD-recognizing integrins. Recent studies have shown that the spike protein does, indeed, interact with αVß3 and α5ß1 integrins, both of which bind to RGD-containing ligands. However, computational studies have suggested that binding between the spike RGD motif and integrins is not favourable, even when unfolding occurs after conformational changes induced by binding to the canonical host entry receptor, angiotensin-converting enzyme 2 (ACE2). Furthermore, non-RGD-binding integrins, such as αx, have been suggested to interact with the SARS-CoV-2 spike protein. Other viral pathogens, such as rotaviruses, have been recorded to bind integrins in an RGD-independent manner to initiate host cell entry. Thus, in order to consider the potential for the SARS-CoV-2 spike protein to bind integrins independent of the RGD sequence, we investigate several factors related to the involvement of integrins in SARS-CoV-2 infection. First, we review changes in integrin expression during SARS-CoV-2 infection to identify which integrins might be of interest. Then, all known non-RGD integrin-binding motifs are collected and mapped to the spike protein receptor-binding domain and analyzed for their 3D availability. Several integrin-binding motifs are shown to exhibit high sequence similarity with solvent accessible regions of the spike receptor-binding domain. Comparisons of these motifs with other betacoronavirus spike proteins, such as SARS-CoV and RaTG13, reveal that some have recently evolved while others are more conserved throughout phylogenetically similar betacoronaviruses. Interestingly, all of the potential integrin-binding motifs, including the RGD sequence, are conserved in one of the known pangolin coronavirus strains. Of note, the most recently recorded mutations in the spike protein receptor-binding domain were found outside of the putative integrin-binding sequences, although several mutations formed inside and close to one motif, in particular, may potentially enhance binding. These data suggest that the SARS-CoV-2 spike protein may interact with integrins independent of the RGD sequence and may help further explain how SARS-CoV-2 and other viruses can evolve to bind to integrins.

What About Promotores? Promotores' Psychosocial Determinants That Influenced the Delivery of a Cervical Cancer Education Intervention to Hispanics.

This study tested whether a cancer education intervention affected promotores' self-efficacy to deliver an intervention to Hispanics and which psychosocial determinants of promotores influenced the number of Hispanic residents reached by promotores in the subsequent education intervention. A quasi-experimental, pre/post-design with a treatment group (no control) assessed differences for promotores (n = 136) before and after exposure to the cancer education intervention. The design also included a cross-sectional evaluation of the number of residents promotores reached with the educational intervention. After being trained, the promotores delivered the intervention to Hispanic residents (n = 1,469). Paired t-tests demonstrated increases in promotores' self-efficacy from pre- to post-intervention. Regression models assessed associations between the numbers of residents reached and select psychosocial determinants of promotores. Age and promotores' years of experience influenced their delivery of a cervical cancer education intervention to Hispanics, but not their delivery of breast or colorectal cancer education interventions. This is the first study to examine which psychosocial determinants influence promotores delivery of cancer education interventions. The outcomes potentially have implications for CHW interventions and training by examining this potential connection between CHWs' psychosocial determinants and intervention outcomes.

Predicted structural mimicry of spike receptor-binding motifs from highly pathogenic human coronaviruses.

Viruses often encode proteins that mimic host proteins in order to facilitate infection. Little work has been done to understand the potential mimicry of the SARS-CoV-2, SARS-CoV, and MERS-CoV spike proteins, particularly the receptor-binding motifs, which could be important in determining tropism and druggability of the virus. Peptide and epitope motifs have been detected on coronavirus spike proteins using sequence homology approaches; however, comparing the three-dimensional shape of the protein has been shown as more informative in predicting mimicry than sequence-based comparisons. Here, we use structural bioinformatics software to characterize potential mimicry of the three coronavirus spike protein receptor-binding motifs. We utilize sequence-independent alignment tools to compare structurally known protein models with the receptor-binding motifs and verify potential mimicked interactions with protein docking simulations. Both human and non-human proteins were returned for all three receptor-binding motifs. For example, all three were similar to several proteins containing EGF-like domains: some of which are endogenous to humans, such as thrombomodulin, and others exogenous, such as Plasmodium falciparum MSP-1. Similarity to human proteins may reveal which pathways the spike protein is co-opting, while analogous non-human proteins may indicate shared host interaction partners and overlapping antibody cross-reactivity. These findings can help guide experimental efforts to further understand potential interactions between human and coronavirus proteins.

Predictors of COVID-19 Preventive Perceptions and Behaviors Among Millennials: Two Cross-sectional Survey Studies.

BACKGROUND: COVID-19 preventive perceptions and behaviors, especially among US millennials, are an important means by which the pandemic can be slowed and negative health outcomes can be averted. OBJECTIVE: This manuscript aims to advance knowledge on COVID-19 preventive perceptions and behaviors and their main predictors, including digital health information-seeking behavior (HISB), political party identification, and COVID-19 testing status. METHODS: Two cross-sectional online surveys of US millennials were conducted from April 10 to 14, 2020 (N=274) (ie, Study 1), and from April 27 to May 7, 2020 (N=1037) (ie, Study 2). In the regression models, dependent variables included preventive behaviors (eg, wearing a face mask and social distancing) as well as four preventive perceptions: severity (ie, a person's conception of the seriousness of COVID-19), susceptibility (ie, a person's conception of the likelihood of being infected with COVID-19), self-efficacy (ie, a person's perception that he or she can wear a face mask and perform social distancing to prevent COVID-19 infection), and response efficacy (ie, a person's perception of whether wearing a face mask and social distancing can prevent COVID-19 infection). Key independent variables included digital HISB for self, digital HISB for another person, political party identification, and COVID-19 testing status. RESULTS: Millennials reported lower levels of perceived susceptibility than the other three preventive perceptions (ie, severity, self-efficacy, and response efficacy), as well as fairly high levels of preventive behaviors. Unlike HISB for another person, digital HISB for self was positively associated with preventive perceptions and behaviors. In Study 1, respondents with higher levels of digital HISB for self had significantly higher perceptions of severity (ß=.22, P<.001), self-efficacy (ß=.15, P=.02), and response efficacy (ß=.25, P<.001) as well as, at nearing significance, higher perceptions of susceptibility (ß=.11, P=.07). In Study 2, respondents with higher levels of digital HISB for self had significantly higher perceptions of severity (ß=.25, P<.001), susceptibility (ß=.14, P<.001), and preventive behaviors (ß=.24, P<.001). Preventive behaviors did not vary significantly according to political party identification, but preventive perceptions did. In Study 1, respondents who identified as being more Republican had significantly lower perceptions of self-efficacy (ß=-.14, P=.02) and response efficacy (ß=-.13, P=.03) and, at nearing significance, lower perceptions of severity (ß=-.10, P=.08) and susceptibility (ß=-.12, P=.06). In Study 2, respondents who identified as being more Republican had significantly lower perceptions of severity (ß=-.08, P=.009). There were mixed effects of COVID-19 testing status on preventive perceptions, with respondents who had tested positive for COVID-19 having significantly higher perceptions of susceptibility in Study 1 (ß=.17, P=.006) and significantly lower perceptions of severity in Study 2 (ß=-.012, P<.001). CONCLUSIONS: As the largest and most digitally savvy generation, US millennials saw COVID-19 as a severe threat, but one that they were less susceptible to. For millennials, digital HISB for self, but not for another person, was critical to the development of preventive perceptions and behaviors.

SARS-CoV-2 3D database: understanding the coronavirus proteome and evaluating possible drug targets.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a rapidly growing infectious disease, widely spread with high mortality rates. Since the release of the SARS-CoV-2 genome sequence in March 2020, there has been an international focus on developing target-based drug discovery, which also requires knowledge of the 3D structure of the proteome. Where there are no experimentally solved structures, our group has created 3D models with coverage of 97.5% and characterized them using state-of-the-art computational approaches. Models of protomers and oligomers, together with predictions of substrate and allosteric binding sites, protein-ligand docking, SARS-CoV-2 protein interactions with human proteins, impacts of mutations, and mapped solved experimental structures are freely available for download. These are implemented in SARS CoV-2 3D, a comprehensive and user-friendly database, available at https://sars3d.com/. This provides essential information for drug discovery, both to evaluate targets and design new potential therapeutics.

Emotions in the time of coronavirus: Antecedents of digital and social media use among Millennials.

Increasingly, people are turning to digital and social media to address health threats. While research has commonly investigated the psychosocial antecedents of digital health information seeking behavior (digital HISB) and social media use (SMU), there has been limited research on the independent effects of emotions and no research on the interactive effects of emotions. In the context of the coronavirus COVID-19 pandemic, this study investigates the affective, personal relevance, and socio-demographic antecedents of digital HISB and SMU, using data from an online survey of U.S. Millennials (N = 1037) in April-May 2020. Linear regression results show the effects of socio-demographic and personal relevance factors. For the independent effects of emotions, fear and sadness were associated with digital HISB; fear, joy, and disgust were associated with high-informational SMU; and joy and disgust were associated with low-informational SMU. Three interactive effects of discrete negative emotions suggest that an increase in one emotion can amplify the effect of another emotion on a measure of digital and social media use. In the fourth interaction of two negative emotions with strong biological regulatory processes, there is evidence that an increase in one emotion can diminish the effect of another emotion. Implications for theory and practice are discussed.

The Role of Enjoyment in a Serious Game for Binge Drinking Prevention: Pretest-Posttest Quasi-Experimental Study.

BACKGROUND: Although binge drinking peaks at age 21 to 25 years, there is limited research on the effects of serious games in this population, as well as on the process by which playing serious games impacts alcohol-related outcomes. Designed with both health behavioral theory and game theory, One Shot is an online serious game that aims to prevent binge drinking. OBJECTIVE: This study utilized a conceptual model for serious video game processes. Using One Shot, the model assessed the following process stages: (1) Alcohol Use Disorders Identification Test-Concise (AUDIT-C); (2) in-game factors of game time and risky alcohol decisions; (3) game enjoyment; and (4) postgame outcomes of intention to drink less and drinking refusal self-efficacy. METHODS: In a one-group pretest-posttest quasi-experimental design, a sample (N=550) of young adults (age 21-25 years) who reported recent binge drinking played the One Shot game. Intention to drink less and drinking refusal self-efficacy were measured at pregame and postgame, with their effects lagged in statistical analysis. Participants were presented with various scenarios in the game that pertained to risky alcohol decisions, which, along with game time, were unobtrusively recorded by the server. A structural equation model (SEM) was used to test the conceptual model, with assessments made to determine if enjoyment mediated the effects of game time and risky alcohol decisions on the 2 postgame alcohol-related outcomes. RESULTS: A well-fitting SEM demonstrated support for the multistep model, with AUDIT-C predicting risky alcohol decisions (ß=.30). Risky alcohol decisions (ß=-.22) and game time (ß=.18) predicted enjoyment, which, in turn, predicted intention to drink less (ß=.21) and drinking refusal self-efficacy (ß=.16). Enjoyment significantly (P<.001) mediated the effects of game time and risky alcohol decision on intention to drink less and drinking refusal self-efficacy. CONCLUSIONS: The results support a conceptual model in which staggered individual and in-game factors influence alcohol-related outcomes. Enjoyment is important for participants' intentions to drink less and beliefs that they can refuse alcohol.

Producing polished prokaryotic pangenomes with the Panaroo pipeline.

Population-level comparisons of prokaryotic genomes must take into account the substantial differences in gene content resulting from horizontal gene transfer, gene duplication and gene loss. However, the automated annotation of prokaryotic genomes is imperfect, and errors due to fragmented assemblies, contamination, diverse gene families and mis-assemblies accumulate over the population, leading to profound consequences when analysing the set of all genes found in a species. Here, we introduce Panaroo, a graph-based pangenome clustering tool that is able to account for many of the sources of error introduced during the annotation of prokaryotic genome assemblies. Panaroo is available at https://github.com/gtonkinhill/panaroo .

Comparison of Surface-Bound and Free-Standing Variations of HKUST-1 MOFs: Effect of Activation and Ammonia Exposure on Morphology, Crystallinity, and Composition.

Metal-organic frameworks (MOFs) are extremely porous, crystalline materials with high surface area for potential use in gas storage, sequestration, and separations. Toward incorporation into structures for these applications, this study compares three variations of surface-bound and free-standing HKUST-1 MOF structures: surface-anchored MOF (surMOF) thin film, drop-cast film, and bulk powder. Herein, effects of HKUST-1 ammonia interaction and framework activation, which is removal of guest molecules via heat, are investigated. Impact on morphology and crystal structure as a function of surface confinement and size variance are examined. Scanning probe microscopy, scanning electron microscopy, powder X-ray diffraction, Fourier-transform infrared spectroscopy, and energy dispersive X-ray spectroscopy monitor changes in morphology and crystal structure, track ammonia uptake, and examine elemental composition. After fabrication, ammonia uptake is observed for all MOF variations, but reveals dramatic morphological and crystal structure changes. However, activation of the framework was found to stabilize morphology. For activated surMOF films, findings demonstrate consistent morphology throughout uptake, removal, and recycling of ammonia over multiple exposures. To understand morphological effects, additional ammonia exposure experiments with controlled post-synthetic solvent adsorbates were conducted utilizing a HKUST-1 standard powder. These findings are foundational for determining the capabilities and limitation of MOF films and powders.

The Outcome Evaluation of a CHW Cancer Prevention Intervention: Testing Individual and Multilevel Predictors Among Hispanics Living Along the Texas-Mexico Border.

This paper evaluates the effectiveness of community health workers/promotores (CHWs) in promoting cancer preventive behaviors in the 2011-2013 Education to Promote Improved Cancer Outcomes (ÉPICO) project. The ÉPICO project utilized CHWs to disseminate cancer education to predominately Spanish-speaking Hispanics living in colonias in the Lower Rio Grande Valley of Texas. The CHWs received training to become Texas-certified CHW instructors and specialized training in message tailoring, and they delivered more than 5000 units of resident education on cancer prevention/detection, treatment, and survivorship for breast, cervical, and colorectal cancer. Using panel data to examine overtime changes in cancer knowledge among Lower Rio Grande Valley residents, the evaluation found significant changes from baseline to both times 1 and 2. Additional individual-level analysis indicated that the increase in resident cancer knowledge was predicted by residents' perceptions of CHW credibility and intention to change their lifestyles. Multilevel analysis also showed that the increase in cancer prevention knowledge among residents was predicted by attributes of the CHWs who taught them. In particular, CHWs with higher education levels had the most impact on residents' increased knowledge over time. Unexpectedly, CHWs with more years of experience were less effective teachers than their early-career counterparts.

Testing the Validity of Campaign Ad Exposure Measures: A Family Planning Media Campaign in Pakistan.

Although prior research has tested the nomological validity of media campaign exposure, including the related comparative validity of some measures, it has not well studied predictive validity or made extensions to other types of media campaign exposure. To help build on research in this area, the current study tested the nomological and predictive validity of 5 ad recall and recognition measures specific to the Touch condom media campaign in Pakistan. Between-effects regression of panel survey data confirmed the nomological validity of each of the 5 measures of Touch ad exposure. In addition, 2 sets of panel regression models (i.e., fixed-effects models and fixed-effects with lag models) confirmed the predictive validity of each of the 5 ad exposure measures. Results on comparative validity were quite similar for nomological and predictive validity, indicating that confirmed ad recall and recognition measures tend to have greater validity than unconfirmed measures.

Smoking Prevention in China: A Content Analysis of an Anti-Smoking Social Media Campaign.

The China Tobacco Control Media Campaign on Sina Weibo is novel in the context of smoking prevention and cessation in China and has not to date been evaluated. This study draws on health behavior theories and dialogic theory in public relations to analyze microblog campaign postings and their relationships with the outcome of online audience engagement. Microblog postings from May 2011 to January 2015 were content analyzed, showing that the most common persuasive content characteristic was perceived risk, followed by subjective norms and self-efficacy. Perceived risk and self-efficacy postings positively influenced online audience engagement, whereas subjective norm postings was a nonsignificant predictor. Postings were more likely to share information than aim to interact with audience members. However, both information sharing and audience interaction postings were positive predictors of online audience engagement. There was also evidence of main and interactive effects of message originality on online audience engagement. The current study has, to the best of our knowledge, broken new ground in 2 regards: (a) using health behavior theories as a basis for analyzing the content of an anti-smoking social media campaign and (b) examining the content of an anti-smoking media campaign of any type in China.

Estimating Causal Effects With Propensity Score Models: An Evaluation of the Touch Condom Media Campaign in Pakistan.

Rapid population growth in Pakistan poses major risks, including those pertinent to public health. In the context of family planning in Pakistan, the current study evaluates the Touch condom media campaign and its effects on condom-related awareness, attitudes, behavioral intention, and behavior. This evaluation relies on 3 waves of panel survey data from men married to women ages 15-49 living in urban and rural areas in Pakistan (N = 1,012): Wave 1 was March 15 to April 7, 2009; Wave 2 was August 10 to August 24, 2009; and Wave 3 was May 1 to June 13, 2010. Analysis of variance provided evidence of improvements in 10 of 11 condom-related outcomes from Wave 1 to Wave 2 and Wave 3. In addition, there was no evidence of outcome decay 1 year after the conclusion of campaign advertising dissemination. To help compensate for violating the assumption of random assignment, propensity score modeling offered evidence of the beneficial effects of confirmed Touch ad recall on each of the 11 outcomes in at least 1 of 3 time-lagged scenarios. By using these different time-lagged scenarios (i.e., from Wave 1 to Wave 2, from Wave 1 to Wave 3, and from Wave 2 to Wave 3), propensity score modeling permitted insights into how the campaign had time-variant effects on the different types of condom-related outcomes, including carryover effects of the media campaign.

Bioanalytical advances in assays for C-reactive protein.

This review presents advances in assays for human C-reactive protein (CRP), the most important biomarker of infection and inflammation for a plethora of diseases and pathophysiological conditions. Routine assays in clinical settings are based on analyzers, enzyme-linked immunosorbent assays and lateral flow assays. However, assays encompassing novel sensing schemes, improved chemistry, signal enhancement, lab-on-a-chip, microfluidics and smartphone detection, have emerged in recent years. The incorporation of immune-transducing chips or sensing interfaces with nanomaterials enables multiplexing analysis of CRP with co-existing biomarkers. However, there are still considerable challenges in the development of rapid diagnostics for both pentameric and monomeric CRP forms.