Designing Epigenome Editors: Considerations of Biochemical and Locus Specificities.

The advent of locus-specific protein recruitment technologies has enabled a new class of studies in chromatin biology. Epigenome editors (EEs) enable biochemical modifications of chromatin at almost any specific endogenous locus. Their locus-specificity unlocks unique information including the functional roles of distinct modifications at specific genomic loci. Given the growing interest in using these tools for biological and translational studies, there are many specific design considerations depending on the scientific question or clinical need. Here, we present and discuss important design considerations and challenges regarding the biochemical and locus specificities of epigenome editors. These include how to: account for the complex biochemical diversity of chromatin; control for potential interdependency of epigenome editors and their resultant modifications; avoid sequestration effects; quantify the locus specificity of epigenome editors; and improve locus-specificity by considering concentration, affinity, avidity, and sequestration effects.

Source terms for benchmarking models of SARS-CoV-2 transmission via aerosols and droplets.

There is ongoing and rapid advancement in approaches to modelling the fate of exhaled particles in different environments relevant to disease transmission. It is important that models are verified by comparison with each other using a common set of input parameters to ensure that model differences can be interpreted in terms of model physics rather than unspecified differences in model input parameters. In this paper, we define parameters necessary for such benchmarking of models of airborne particles exhaled by humans and transported in the environment during breathing and speaking.

Viral load of SARS-CoV-2 in droplets and bioaerosols directly captured during breathing, speaking and coughing.

Determining the viral load and infectivity of SARS-CoV-2 in macroscopic respiratory droplets, bioaerosols, and other bodily fluids and secretions is important for identifying transmission modes, assessing risks and informing public health guidelines. Here we show that viral load of SARS-CoV-2 Ribonucleic Acid (RNA) in participants' naso-pharyngeal (NP) swabs positively correlated with RNA viral load they emitted in both droplets >10 [Formula: see text] and bioaerosols <10 [Formula: see text] directly captured during the combined expiratory activities of breathing, speaking and coughing using a standardized protocol, although the NP swabs had [Formula: see text] 10[Formula: see text] more RNA on average. By identifying highly-infectious individuals (maximum of 18,000 PFU/mL in NP), we retrieved higher numbers of SARS-CoV-2 RNA gene copies in bioaerosol samples (maximum of 4.8[Formula: see text] gene copies/mL and minimum cycle threshold of 26.2) relative to other studies. However, all attempts to identify infectious virus in size-segregated droplets and bioaerosols were negative by plaque assay (0 of 58). This outcome is partly attributed to the insufficient amount of viral material in each sample (as indicated by SARS-CoV-2 gene copies) or may indicate no infectious virus was present in such samples, although other possible factors are identified.

Ionomers From Kraft Lignin for Renewable Energy Applications.

Converting industrial/agricultural lignin-rich wastes to efficient, cost-effective materials for electrochemical devices (e.g., fuel cells) can aid in both bio- and energy economy. A major limitation of fuel cells is the weak ion conductivity within the ~2-30-nm thick, ion-conducting polymer (ionomer)-based catalyst-binder layer over electrodes. Here, we strategically sulfonated kraft lignin (a by-product of pulp and paper industries) to design ionomers with varied ion exchange capacities (IECs) (LS x; x = IEC) that can potentially overcome this interfacial ion conduction limitation. We measured the ion conductivity, water uptake, ionic domain characteristics, density, and predicted the water mobility/stiffness of Nafion, LS 1.6, and LS 3.1 in submicron-thick hydrated films. LS 1.6 showed ion conductivity an order of magnitude higher than Nafion and LS 3.1 in films with similar thickness. The ion conductivity of these films was not correlated to their water uptake and IECs. Within the three-dimensional, less dense, branched architecture of LS 1.6 macromolecules, the -SO3H and -OH groups are in close proximity, which likely facilitated the formation of larger ionic domains having highly mobile water molecules. As compared to LS 1.6, LS 3.1 showed a higher glass transition temperature and film stiffness at dry state, which sustained during humidification. On the contrary, Nafion stiffened significantly upon humidification. The smaller ionic cluster within stiff LS 3.1 and Nafion films thus led to ion conductivity lower than LS 1.6. Since LS x ionomers (unlike commercial lignosulfonate) are not water soluble, they are suitable for low-temperature, water-mediated ion conduction in submicron-thick films.

Cationic π-Conjugated Polyelectrolyte Shows Antimicrobial Activity by Causing Lipid Loss and Lowering Elastic Modulus of Bacteria.

Cationic, π-conjugated oligo-/polyelectrolytes (CCOEs/CCPEs) have shown great potential as antimicrobial materials to fight against antibiotic resistance. In this work, we treated wild-type and ampicillin-resistant (amp-resistant) Escherichia coli (E. coli) with a promising cationic, π-conjugated polyelectrolyte (P1) with a phenylene-based backbone and investigated the resulting morphological, mechanical, and compositional changes of the outer membrane of bacteria in great detail. The cationic quaternary amine groups of P1 led to electrostatic interactions with negatively charged moieties within the outer membrane of bacteria. Using atomic force microscopy (AFM), high-resolution transmission electron microscopy (TEM), we showed that due to this treatment, the bacterial outer membrane became rougher, decreased in stiffness/elastic modulus (AFM nanoindentation), formed blebs, and released vesicles near the cells. These evidences, in addition to increased staining of the P1-treated cell membrane by lipophilic dye Nile Red (confocal laser scanning microscopy (CLSM)), suggested loosening/disruption of packing of the outer cell envelope and release and exposure of lipid-based components. Lipidomics and fatty acid analysis confirmed a significant loss of phosphate-based outer membrane lipids and fatty acids, some of which are critically needed to maintain cell wall integrity and mechanical strength. Lipidomics and UV-vis analysis also confirmed that the extracellular vesicles released upon treatment (AFM) are composed of lipids and cationic P1. Such surface alterations (vesicle/bleb formation) and release of lipids/fatty acids upon treatment were effective enough to inhibit further growth of E. coli cells without completely disintegrating the cells and have been known as a defense mechanism of the cells against cationic antimicrobial agents.

Comprehensive characterization of mainstream marijuana and tobacco smoke.

Recent increases in marijuana use and legalization without adequate knowledge of the risks necessitate the characterization of the billions of nanoparticles contained in each puff of smoke. Tobacco smoke offers a benchmark given that it has been extensively studied. Tobacco and marijuana smoke particles are quantitatively similar in volatility, shape, density and number concentration, albeit with differences in size, total mass and chemical composition. Particles from marijuana smoke are on average 29% larger in mobility diameter than particles from tobacco smoke and contain 3.4× more total mass. New measurements of semi-volatile fractions determine over 97% of the mass and volume of the particles from either smoke source are comprised of semi-volatile compounds. For tobacco and marijuana smoke, respectively, 4350 and 2575 different compounds are detected, of which, 670 and 536 (231 in common) are tentatively identified, and of these, 173 and 110 different compounds (69 in common) are known to cause negative health effects through carcinogenic, mutagenic, teratogenic, or other toxic mechanisms. This study demonstrates striking similarities between marijuana and tobacco smoke in terms of their physical and chemical properties.

A Simple Method for Measuring Fine-to-Ultrafine Aerosols Using Bipolar Charge Equilibrium.

Low-cost methods for measuring airborne microparticles and nanoparticles (aerosols) have remained elusive despite the increasing concern of health impacts from ambient, urban, and indoor sources. While bipolar ion sources are common in smoke alarms, this work is the first to exploit the mean charge on an aerosol resulting from a bipolar charge equilibrium and establish experimentally its correlation to properties of the aerosol particle size distribution. The net current produced from this mean particle charge is demonstrated to be linearly proportional to the product of the mean particle diameter and total number concentration (i ∼ Nd̅) for two bipolar ion sources (85Kr and 241Am). This conclusion is supported by simple equations derived from well-established bipolar charging theory. The theory predicts that the mean charge on the aerosol particles reaches an equilibrium, which, importantly, is independent of the concentration of charging ions. Furthermore, in situ measurements of a roadside aerosol demonstrate that the sensing method yields results in good agreement (R2 = 0.979) with existing portable and laboratory-grade aerosol instruments. The simplicity, stability, and cost of the bipolar ion source overcome challenges of other portable sensors, increasing the feasibility of widespread sensor deployment to monitor ultrafine particle characteristics, which are relevant to lung deposition and by extension, human health.

Endothelial cell Pannexin1 modulates severity of ischemic stroke by regulating cerebral inflammation and myogenic tone.

Ischemic stroke is a leading cause of morbidity and mortality in the US; however, there currently exists only one effective acute pharmacological therapeutic intervention. Purinergic signaling has been shown to regulate vascular function and pathological processes, including inflammation and arterial myogenic reactivity, and plays a role in ischemic stroke outcome. Purinergic signaling requires extracellular purines; however, the mechanism of purine release from cells is unclear. Pannexin1 (Panx1) channels are potentially novel purine release channels expressed throughout the vascular tree that couples regulated purine release with purinergic signaling. Therefore, we examined the role of smooth muscle and endothelial cell Panx1, using conditional cell type-specific transgenic mice, in cerebral ischemia/reperfusion injury outcomes. Deletion of endothelial cell Panx1, but not smooth muscle cell Panx1, significantly reduced cerebral infarct volume after ischemia/reperfusion. Infiltration of leukocytes into brain tissue and development of cerebral myogenic tone were both significantly reduced when mice lacked endothelial Panx1. Panx1 regulation of myogenic tone was unique to the cerebral circulation, as mesenteric myogenic reactivity and blood pressure were independent of endothelial Panx1. Overall, deletion of endothelial Panx1 mitigated cerebral ischemic injury by reducing inflammation and myogenic tone development, indicating that endothelial Panx1 is a possible novel target for therapeutic intervention of ischemic stroke.

Growth Dynamics of Patient-Provider Internet Communication: Trend Analysis Using the Health Information National Trends Survey (2003 to 2013).

BACKGROUND: Communication is key in chronic disease management, and the internet has altered the manner in which patients and providers can exchange information. Adoption of secure messaging differs among patients due to the digital divide that keeps some populations from having effective access to online resources. OBJECTIVE: This study aimed to examine the current state of online patient-provider communication, exploring trends over time in the use of online patient-provider communication tools. METHODS: A 3-part analytic process was used to study the following: (1) reanalysis, (2) close replication across years, and (3) trend analysis extension. During the reanalysis stage, the publicly available Health Information National Trends Survey (HINTS) 1 and 2 data were used with the goal of identifying the precise analytic methodology used in a prior study, published in 2007. The original analysis was extended to add 3 additional data years (ie, 2008, 2011, and 2013) using the original analytical approach with the purpose of identifying trends over time. Multivariate logistic regression was used to analyze pooled data across all years, with year as an added predictor, in addition to a model for each individual data year. RESULTS: The odds of internet users to communicate online with health care providers was significantly and increasingly higher year-over-year, starting in 2003 (2005: odds ratio [OR] 1.31, 95% CI 1.03-1.68; 2008: OR 2.14, 95% CI 1.76-2.59; 2011: OR 2.92, 95% CI 2.33-3.66; and 2013: OR 5.77; 95% CI 4.62-7.20). Statistically significant socio-economic factors found to be associated with internet users communicating online with providers included age, having health insurance, having a history of cancer, and living in an urban area of residence. CONCLUSIONS: The proportion of internet users communicating online with their health care providers has significantly increased since 2003. Although these trends are encouraging, access challenges still exist for some groups, potentially giving rise to a new set of health disparities related to communication.

Trends in E-Cigarette Awareness and Perceived Harmfulness in the U.S.

INTRODUCTION: Electronic cigarettes (e-cigarettes) are gaining in popularity as an alternative to regular cigarettes, as they are viewed as potentially less harmful. However, it remains unclear how awareness about e-cigarettes is permeating through the general U.S. POPULATION: This study seeks to extend previous research and examine trends in e-cigarette awareness and perceived harmfulness, and their association with smoking-cessation efforts. METHODS: Data from three cycles (2012, 2013, and 2014) of the Health Information National Trends Survey were combined into a single data set. Controlling for survey year, multivariate logit models were used to determine the association between demographic characteristics and e-cigarette awareness, perceived harmfulness, quit attempts, and quit intentions. Data were analyzed in 2015. RESULTS: Awareness of e-cigarettes increased from 77.1% in 2012 to 94.3% in 2014. Controlling for demographic characteristics, e-cigarette awareness significantly increased in both 2013 and 2014, relative to 2012. Perception that e-cigarettes were less harmful than regular cigarettes declined from 50.7% in 2012 to 43.1% in 2014. Among smokers, no relationship was observed between e-cigarette awareness and past-year quit attempts or quit intentions, but those that viewed e-cigarettes as less harmful were less likely to have a past-year quit attempt. CONCLUSIONS: These analyses reveal a continued increase in overall public awareness of e-cigarettes and shifting harm perceptions relative to regular cigarettes. New regulatory oversight by the U.S. Food and Drug Administration may have major effects on both dimensions, which are worth continued monitoring.

Low Reynolds number Couette flow facility for drag measurements.

For this study a new low Reynolds number Couette facility was constructed to investigate surface drag. In this facility, mineral oil was used as the working fluid to increase the shear stress across the surface of the experimental models. A mounted conveyor inside a tank creates a flow above which an experimental model of a flat plate was suspended. The experimental plate was attached to linear bearings on a slide system that connects to a force gauge used to measure the drag. Within the gap between the model and moving belt a Couette flow with a linear velocity profile was created. Digital particle image velocimetry was used to confirm the velocity profile. The drag measurements agreed within 5% of the theoretically predicted Couette flow value.

Evolution of resource competition between mutually dependent digital organisms.

We study the emergence and dynamics of competing strains of digital organisms in a world with two depletable resources. Consumption of one resource produces the other resource as a by-product, and vice versa. As a consequence, two types of mutually dependent organisms emerge that each prey on the waste product of the other. In the absence of mutations, that is, in a purely ecological setting, the abundances of the two types of organisms display a wide range of different types of oscillations, from regular oscillations with large amplitude to irregular oscillations with amplitudes ranging from small to large. In this regime, time-averaged abundance levels seem to be controlled by the relative fitness of the organisms in the absence of resources. Under mutational pressure, on the other hand, populations evolve that seem to avoid the oscillations of intermediate to large amplitudes. In this case, the relative fitness of the organisms in the presence of resources plays an important role in the time-averaged abundance levels as well.