Ablation of Endothelial TRPV4 Channels Alters the Dynamic Ca2+ Signaling Profile in Mouse Carotid Arteries.

Transient receptor potential vanilloid 4 channels (TRPV4) are pivotal regulators of vascular homeostasis. Altered TRPV4 signaling has recently been implicated in various cardiovascular diseases, including hypertension and atherosclerosis. These versatile nonselective cation channels increase endothelial Ca2+ influx in response to various stimuli including shear stress and G protein-coupled receptor (GPCR) activation. Recent findings suggest TRPV4 channels produce localized Ca2+ transients at the endothelial cell plasma membrane that may allow targeted effector recruitment and promote large-scale Ca2+ events via release from internal stores (endoplasmic reticulum). However, the specific impact of TRPV4 channels on Ca2+ signaling in the intact arterial intima remains unknown. In the current study, we employ an endothelium-specific TRPV4 knockout mouse model (ecTRPV4-/-) to identify and characterize TRPV4-dependent endothelial Ca2+ dynamics. We find that carotid arteries from both ecTRPV4-/- and WT mice exhibit a range of basal and acetylcholine (ACh)-induced Ca2+ dynamics, similar in net frequency. Analysis of discrete Ca2+ event parameters (amplitude, duration, and spread) and event composite values reveals that while ecTRPV4-/- artery endothelium predominantly produces large Ca2+ events comparable to and in excess of those produced by WT endothelium, they are deficient in a particular population of small events, under both basal and ACh-stimulated conditions. These findings support the concept that TRPV4 channels are responsible for generating a distinct population of focal Ca2+ transients in the intact arterial endothelium, likely underlying their essential role in vascular homeostasis.

Hyperspectral imaging fluorescence excitation scanning spectral characteristics of remodeled mouse arteries.

Coronary artery disease (CAD), or atherosclerosis, is responsible for nearly a third of all American deaths annually. Detection of plaques and differentiation of plaque stage remains a complicating factor for treatment. Classification of plaque before significant blockage or rupture could inform clinical decisions and prevent mortality. Current detection methods are either nonspecific, slow, or require the use of potentially harmful contrast agents. Recent advances in hyperspectral imaging could be used to detect changes in the autofluorescence of arteries associated with vessel remodeling and subsequent plaque formation and could detect and classify existing lesions. Here, we present data comparing spectral image characteristics of a mouse model designed to undergo vessel remodeling. C57Bl/6 mice underwent ligation of three of four caudal branches of the left common carotid artery (left external carotid, internal carotid, and occipital artery) with the superior thyroid artery left intact under IACUC approved protocol. Vessels were harvested at a variety of timepoints to compare degrees of remodeling, including 4 weeks and 5 months post-surgery. Immediately following harvest, vessels were prepared by longitudinal opening to expose the luminal surface to a 20X objective. A custom inverted microscope (TE-2000, Nikon Instruments) with a Xe arc lamp and thin film tunable filter arrary (Versachrome, Semrock, Inc.) were used to achieve spectral imaging. Excitation scans utilized wavelengths between 340 nm and 550 nm in 5 nm increments. Hyperspectral data were generated and analyzed with custom Matlab scripts and visualized in ENVI. Preliminary data suggest consistent spectral features associated with control and remodeled vessels.

Implementation of a Medicare Plan Selection Assistance Program Through a Community Partnership.

Objective. To describe the implementation and outcomes of a sustainable Medicare Plan Selection Assistance Program conducted through a partnership between Auburn University Harrison School of Pharmacy (AU) and the Alabama State Health Insurance and Assistance Program (SHIP) since 2013. Methods. The program's goal is to assist Medicare beneficiaries in Medicare Part D plan selection. Reported outcomes included Medicare beneficiaries' plan cost savings and satisfaction, and pharmacy students' self-reported changes in knowledge and attitudes. Results. Each year, more than 80 pharmacy students assist more than 120 beneficiaries; at least 10 events are held covering 6-10 Alabama counties. On average, Medicare beneficiaries had a projected savings of $278.71 (2013), $1,081.66 (2014), $842.84 (2015), and $1,382.90 (2016) after enrolling in a new plan, and most students reported perceived increased ability to help beneficiaries select the most appropriate Medicare Part D plan. Conclusion. The program produced positive outcomes for both beneficiaries and students. Other pharmacy schools may consider partnering with their State Health Insurance and Assistance Program to deliver a similar program to benefit their students and Medicare beneficiaries.

Pharmacists' Attitudes and Perceived Barriers to Human Papillomavirus (HPV) Vaccination Services.

Use of non-traditional settings such as community pharmacies has been suggested to increase human papillomavirus (HPV) vaccination uptake and completion rates. The objectives of this study were to explore HPV vaccination services and strategies employed by pharmacies to increase HPV vaccine uptake, pharmacists' attitudes towards the HPV vaccine, and pharmacists' perceived barriers to providing HPV vaccination services in community pharmacies. A pre-piloted mail survey was sent to 350 randomly selected community pharmacies in Alabama in 2014. Measures included types of vaccines administered and marketing/recommendation strategies, pharmacists' attitudes towards the HPV vaccine, and perceived system and parental barriers. Data analysis largely took the form of descriptive statistics. 154 pharmacists completed the survey (response rate = 44%). The majority believed vaccination is the best protection against cervical cancer (85.3%), HPV is a serious threat to health for girls (78.8%) and boys (55.6%), and children should not wait until they are sexually active to be vaccinated (80.1%). Perceived system barriers included insufficient patient demand (56.5%), insurance plans not covering vaccination cost (54.8%), and vaccine expiration before use (54.1%). Respondents also perceived parents to have inadequate education and understanding about HPV infection (86.6%) and vaccine safety (78.7%). Pharmacists have positive perceptions regarding the HPV vaccine. Barriers related to system factors and perceived parental concerns must be overcome to increase pharmacist involvement in HPV vaccinations.

Parental acceptance of human papillomavirus vaccinations and community pharmacies as vaccination settings: A qualitative study in Alabama.

PURPOSE: To determine parents' knowledge and attitudes regarding human papillomavirus (HPV) vaccinations in their adolescent children and to describe parents' perceptions of adolescent vaccinations in community pharmacies. METHODS: In-depth interviews were completed with parents or guardians of children ages 11-17 years from Alabama's Lee and Macon counties. One-hour long, open-ended telephonic or in-person interviews were conducted until the saturation point was reached. Using ATLAS.ti software and thematic analysis, interview transcripts were coded to identify themes. RESULTS: Twenty-six parents were interviewed, most of whom were female (80.8%) and white (50%). A total of 12 themes were identified. First, two themes emerged regarding elements facilitating children's HPV vaccination, the most common being positive perception of the HPV vaccine. Second, elements hindering children's vaccination contained seven themes, the top one being lack of correct or complete information about the HPV vaccine. The last topic involved acceptance/rejection of community pharmacies as vaccination settings, and the most frequently cited theme was concern about pharmacists' clinical training. CONCLUSIONS: Physician-to-parent vaccine education is important, and assurances of adequate pharmacy immunization training will ease parents' fears and allow pharmacists to better serve adolescents, especially those who do not see physicians regularly.

How Do Pharmacists Assist Medicare Beneficiaries with Limited Income? A Cross-Sectional Study of Community Pharmacies in Alabama.

BACKGROUND: Many Medicare beneficiaries have limited income and report problems paying for their medications. Programs are available to assist these low-income individuals. However, these programs are underused because of lack of general awareness and perceived complexity of program applications. OBJECTIVES: To (a) determine the frequency of encounters by pharmacists with Medicare beneficiaries who cannot afford prescription drugs; (b) identify strategies that pharmacists use to assist Medicare beneficiaries who cannot afford prescription drugs; and (c) explore what pharmacists know about programs for Medicare beneficiaries with limited income. METHODS: This study used a mixed-mode survey of 350 randomly sampled community pharmacies located in 32 counties in Alabama with a high proportion of Medicare beneficiaries who were potentially eligible for low-income subsidy programs. Measures included frequency of encounters by pharmacists with Medicare beneficiaries who could not afford their medications, strategies used to assist Medicare beneficiaries, and pharmacists' knowledge of programs for Medicare beneficiaries with limited income. RESULTS: Of 350 surveys sent, 12 were nondeliverable, and 151 were completed (response rate=44.6%). About 50% of respondents reported encountering Medicare beneficiaries who could not afford their medications at least weekly. Various strategies were reported, including refiling claims that were previously denied every day (40.7%), contacting insurance companies at least once per week (43.2%), and loaning medications at least 2-3 times per month (29.1%). Only 12.6% reported referring beneficiaries to the Aging and Disability Resource Centers (ADRCs) to assess eligibility for limited-income programs. When asked about programs for beneficiaries with limited income, the answers were predominantly "don't know for sure." CONCLUSIONS: Several strategies were used by pharmacists in an attempt to help limited-income Medicare beneficiaries obtain their medications. Lack of knowledge about financial assistance programs for limited-income individuals and the role of ADRCs in helping to screen individuals for benefits and complete applications warrants immediate attention. Improving pharmacists' knowledge on this topic may be an effective mechanism for providing a long-term solution for their patients. DISCLOSURES: This study was funded by the Alabama Department of Senior Services. The sponsor played no active role in the design, methods, data collection, analysis, or preparation of this manuscript. The authors have nothing to disclose. This paper was presented at the American Pharmacists Association meeting in Baltimore, Maryland, in 2016. Study concept and design were contributed by Westrick. Hastings and McFarland were responsible for data collection, along with Westrick. Data interpretation was performed by Westrick, L. Hohmann, and Hastings, with assistance from McFarland. All authors were involved with manuscript preparation.

Characterization and Mechanism for the Protection of Photolytic Decomposition of N-Halamine Siloxane Coatings by Titanium Dioxide.

N-Halamine antibacterial materials have superior inactivation activities due to oxidative chlorine species. However, N-Cl bonds and bonds between N-halamine and substrates often decompose rapidly under UV irradiation, leading to unrecoverable loss of antimicrobial activity. In this study, titanium dioxide was covalently bonded onto N-halamine siloxane poly[5,5-dimethyl-3-(3'-triethoxysilylpropyl)hydantoin] (PSPH) via a sol-gel process. Experimental testing of the chlorinated cotton fabrics treated with TiO2/PSPH demonstrated that the residual oxidative chlorine in cotton-TiO2/PSPH-Cl was still effective for inactivating bacteria after 50 washing cycles and under UV light irradiation for 24 h. Quantum mechanical calculations found that TiO2 improves the UV stability of the PSPH-Cl system by increasing the activation barrier of the C-Si scission reaction responsible for the loss of the biocidal hydantoin moiety. SEM, XPS and FTIR spectra were used to characterize the coated cotton samples. Cotton-TiO2/PSPH-Cl samples exhibited good antibacterial activity against Staphylococcus aureus (ATCC 6538) and Escherichia coli O157:H7 (ATCC 43895). The storage stability and washing stability of treated cotton fabrics were also investigated.

Benchmarking Continuum Solvent Models for Keto-Enol Tautomerizations.

Experimental free energies of tautomerization, ΔGT, were used to benchmark the gas-phase predictions of 17 different quantum mechanical methods and eight basis sets for seven keto-enol tautomer pairs dominated by their enolic form. The G4 method and M06/6-31+G(d,p) yielded the most accurate results, with mean absolute errors (MAE's) of 0.95 and 0.71 kcal/mol, respectively. Using these two theory levels, the solution-phase ΔGT values for 23 unique tautomer pairs composed of aliphatic ketones, ß-dicarbonyls, and heterocycles were computed in multiple protic and aprotic solvents. The continuum solvation models, namely, polarizable continuum model (PCM), polarizable conductor calculation model (CPCM), and universal solvation model (SMD), gave relatively similar MAE's of ∼1.6-1.7 kcal/mol for G4 and ∼1.9-2.0 kcal/mol with M06/6-31+G(d,p). Partitioning the tautomer pairs into their respective molecular types, that is, aliphatic ketones, ß-dicarbonyls, and heterocycles, and separating out the aqueous versus nonaqueous results finds G4/PCM utilizing the UA0 cavity to be the overall most accurate combination. Free energies of activation, ΔG(‡), for the base-catalyzed keto-enol interconversion of 2-nitrocyclohexanone were also computed using six bases and five solvents. The M06/6-31+G(d,p) reproduced the ΔG(‡) with MAE's of 1.5 and 1.8 kcal/mol using CPCM and SMD, respectively, for all combinations of base and solvent. That specific enolization was previously proposed to proceed via a concerted mechanism in less polar solvents but shift to a stepwise mechanism in more polar solvents. However, the current calculations suggest that the stepwise mechanism operates in all solvents.