News coverage about aspirin as a countervailing force against low-dose aspirin campaign promotion.

Organized health promotion efforts sometimes compete with news media, social media, and other sources when providing recommendations for healthy behavior. In recent years, patients have faced a complicated information environment regarding aspirin use as a prevention tool for heart health. We explored the possibility that campaign promotion of low-dose aspirin use might have been undermined by news coverage in the USA detailing controversies regarding aspirin use. Using time series data on low-dose aspirin sales in Minnesota, USA, we assessed whether news coverage of aspirin or audience engagement with the Ask About Aspirin campaign website predicted subsequent changes in low-dose aspirin sales, over and above any secular trend. News coverage predicted actual low-dose aspirin purchases whereas exposure to a state-level campaign did not. While a campaign effort to encourage people at risk to discuss low-dose aspirin use with their health care providers did not generate substantive changes in low-dose aspirin tablet sales in the areas of Minnesota monitored for this study, past news coverage about aspirin use, including news about negative side effects, may have suppressed low-dose aspirin sales during this same period. The extent of news coverage about aspirin and heart health had a negative effect on tablet sales recorded in greater Minnesota approximately a month later in an ARIMA time series model, coefficient = -.014, t = -2.33, p = .02. Presented evidence of news coverage effect suggests health campaign assessment should consider trends in the public information environment as potential countervailing forces.

News coverage and online advertising effects on patient-led search for aspirin, heart health, and stroke information and educational tool use.

OBJECTIVE: Using indicators of campaign effort and relevant news stories, we sought to predict two patterns of patient behavior regarding information about aspirin and heart health: patient use of a campaign web tool to determine whether they should talk with a physician about using aspirin and patient searches for information about aspirin and the heart. METHODS: We used ARIMA modeling to predict two time series as a function of independent variables. RESULTS: We found significant prediction of time series in both models, but campaign expenditure only predicted use of a campaign web tool whereas weekly news stories predicted online searches regarding aspirin and the heart originating from Minnesota. CONCLUSION: Patient information engagement is a function of information salience at least in part. Campaign advertising expenditure can prompt audience use of campaign tools but news coverage also operates as an important force on patient search behavior. PRACTICE IMPLICATIONS: Health promotion professionals charged with reaching patients with heart health and stroke prevention messages should monitor news coverage as a potential complementary or rival force while at the same time promoting campaign-related information online.

A deep learning-based algorithm for 2-D cell segmentation in microscopy images.

BACKGROUND: Automatic and reliable characterization of cells in cell cultures is key to several applications such as cancer research and drug discovery. Given the recent advances in light microscopy and the need for accurate and high-throughput analysis of cells, automated algorithms have been developed for segmenting and analyzing the cells in microscopy images. Nevertheless, accurate, generic and robust whole-cell segmentation is still a persisting need to precisely quantify its morphological properties, phenotypes and sub-cellular dynamics. RESULTS: We present a single-channel whole cell segmentation algorithm. We use markers that stain the whole cell, but with less staining in the nucleus, and without using a separate nuclear stain. We show the utility of our approach in microscopy images of cell cultures in a wide variety of conditions. Our algorithm uses a deep learning approach to learn and predict locations of the cells and their nuclei, and combines that with thresholding and watershed-based segmentation. We trained and validated our approach using different sets of images, containing cells stained with various markers and imaged at different magnifications. Our approach achieved a 86% similarity to ground truth segmentation when identifying and separating cells. CONCLUSIONS: The proposed algorithm is able to automatically segment cells from single channel images using a variety of markers and magnifications.

Effects of conscious sedation on patient recall of anxiety and pain after oral surgery.

OBJECTIVE: This study examined the effect of conscious ("moderate") sedation with amnestic effects and local anesthetic, versus local anesthetic alone, on recall of pain and anxiety related to surgical tooth extraction. Greater anxiety and pain were hypothesized in the local anesthesia-alone group. STUDY DESIGN: Patients undergoing tooth extraction, receiving moderate sedation plus local anesthetic (n = 27) or local anesthetic alone (n = 27), were assessed on trait dental anxiety, preextraction state pain and anxiety, anticipated pain and anxiety, and 1-month recall of pain and anxiety. RESULTS: Patients with moderate sedation, compared with those administered only local anesthetic, recalled less procedural pain and anxiety after 1 month. The local anesthetic-alone group reported more preextraction pain and anticipated more procedural anxiety. CONCLUSIONS: Moderate sedation had the desired effect of lower recalled pain and anxiety associated with extraction, even 1 month later. Anticipating moderate sedation also prompts expectation of less anxiety during the procedure.

Validation of a high-content screening assay using whole-well imaging of transformed phenotypes.

Automated microscopy was introduced two decades ago and has become an integral part of the discovery process as a high-content screening platform with noticeable challenges in executing cell-based assays. It would be of interest to use it to screen for reversers of a transformed cell phenotype. In this report, we present data obtained from an optimized assay that identifies compounds that reverse a transformed phenotype induced in NIH-3T3 cells by expressing a novel oncogene, KP, resulting from fusion between platelet derived growth factor receptor alpha (PDGFRα) and kinase insert domain receptor (KDR), that was identified in human glioblastoma. Initial image acquisitions using multiple tiles per well were found to be insufficient as to accurately image and quantify the clusters; whole-well imaging, performed on the IN Cell Analyzer 2000, while still two-dimensional imaging, was found to accurately image and quantify clusters, due largely to the inherent variability of their size and well location. The resulting assay exhibited a Z' value of 0.79 and a signal-to-noise ratio of 15, and it was validated against known effectors and shown to identify only PDGFRα inhibitors, and then tested in a pilot screen against a library of 58 known inhibitors identifying mostly PDGFRα inhibitors as reversers of the KP induced transformed phenotype. In conclusion, our optimized and validated assay using whole-well imaging is robust and sensitive in identifying compounds that reverse the transformed phenotype induced by KP with a broader applicability to other cell-based assays that are challenging in HTS against chemical and RNAi libraries.

Non-small-cell lung cancers with kinase domain mutations in the epidermal growth factor receptor are sensitive to ionizing radiation.

Non-small cell lung cancers (NSCLCs) bearing mutations in the tyrosine kinase domain (TKD) of the epidermal growth factor receptor (EGFR) often exhibit dramatic sensitivity to the EGFR tyrosine kinase inhibitors gefitinib and erlotinib. Ionizing radiation (IR) is frequently used in the treatment of NSCLC, but little is known how lung tumor-acquired EGFR mutations affect responses to IR. Because this is of great clinical importance, we investigated and found that clonogenic survival of mutant EGFR NSCLCs in response to IR was reduced 500- to 1,000-fold compared with wild-type (WT) EGFR NSCLCs. Exogenous expression of either the L858R point mutant or the DeltaE746-E750 deletion mutant form of EGFR in immortalized human bronchial epithelial cells, p53 WT NSCLC (A549), or p53-null NSCLC (NCI-H1299) resulted in dramatically increased sensitivity to IR. We show that the majority of mutant EGFR NSCLCs, including those that contain the secondary gefitinib resistance T790M mutation, exhibit characteristics consistent with a radiosensitive phenotype, which include delayed DNA repair kinetics, defective IR-induced arrest in DNA synthesis or mitosis, and pronounced increases in apoptosis or micronuclei. Thus, understanding how activating mutations in the TKD domain of EGFR contribute to radiosensitivity should provide new insight into effective treatment of NSCLC with radiotherapy and perhaps avoid emergence of single agent drug resistance.

Feedstock optimization of in-vessel food waste composting systems for inactivation of pathogenic microorganisms.

An optimum composting recipe was investigated to reduce pathogenic microorganisms in a forced-aerated in-vessel system (55 liters). The feedstocks used for in-vessel composting were food waste, cow manure, and bulking materials (wood shavings and mulch hay). A statistical extreme vertices mixture design method was used to design the composting experiments and analyze the collected data. Each mixture (nine total) was replicated randomly three times. Temperature was monitored as an indicator of the efficiency of the composting experiments. The maximum temperature values of the mixtures were used as a response for both extreme vertices mixture design and statistical analyses. Chemical changes (moisture content, carbon/nitrogen ratio, volatile solids, and pH) and reductions of indicator (fecal coliforms and fecal streptococci) and pathogenic microorganisms (Salmonella and Escherichia coli O157:H7) were measured by the most-probable-number method before and after a 12-day composting period. Maximum temperatures for the tested compost mixtures were in the range of 37.0 to 54.7 degrees C. Extreme vertices mixture design analysis of the surface plot suggested an optimum mixture containing 50% food waste, 40% manure, and 10% bulking agents. This optimum mixture achieved maximum temperatures of 54.7 to 56.6 degrees C for about 3.3 days. The total reduction of Salmonella and E. coli O157:H7 were 92.3%, whereas fecal coliforms and fecal streptococci reductions were lower (59.3 and 27.1%, respectively). Future study is neededto evaluate the extreme vertices mixture design method for optimization of large-scale composting.

Development of an assay suitable for high-throughput screening to measure matrix metalloprotease activity.

MMPs, part of a family of enzymes with >35 known members, play an important role in tissue remodeling and repair, in the biology of neoplasia, and during development. Hydroxamic and carboxylic acid inhibitors of these proteases have long been available, but their specificities are poor and there still exists a desire to find novel chemical structures, which could be modified to optimize specificity and biocompatibility. Established methods for measuring MMP activity are based on the cleavage of MCA-PLGL-A2pr(DNP)-AR, which provides a prompt fluorescent signal when cleaved; however, its absorption/emission properties (325/400 nm) are not best suited for HTS assays. We describe an HTS-compatible method using the peptide substrate PLGLAARK, labeled at N- and C-termini with CyDye fluors Cy3 and Cy5Q, respectively, which is cleavable by MMP-1, -2, -3, -7, -9, and -13. HTS assays for MMP-13 and MMP-9 inhibitors were set up in approximately 20 microl in 384-well plates as a prompt fluorescence readout (excitation/emission = 540/570 nm) using the LEADseeker homogenous imaging system. These assays yielded IC(50) values comparable to standard methods, but with a faster, very sensitive, and normalized readout, thus conserving compound, enzyme (approximately 1.5 ng/well), and time (20 s read/plate). Data quality (Z' approximately 0.9) was such that hit-picking to -25% change in primary screening could be performed with confidence, and the subsequent rate of confirmation and validation in IC(50) determinations of the picked compounds was >60%. Parallel screening of related proteases also permitted immediate specificity comparisons, including evaluation of inactive or weakly active compounds.