Evidence-based diabetes prevention and control programs and policies in local health departments

PURPOSE: The purpose of this study is to: (1) assess implementation of evidence-based programs and policies (EBPPs) related to diabetes prevention and control in local health departments; (2) assess feasibility of non-implemented diabetes prevention and control EBPPs; and (3) examine individual- and organizational-level factors associated with implementation of diabetes prevention and control EBPPs. METHODS: An online survey was administered in January 2015 to key representatives of all local health departments in Missouri. Descriptive statistics were used to describe implementation and perceived feasibility of 20 diabetes prevention and control EBPPs. Logistic regression was used to examine the association between individual and organizational factors and diabetes prevention and control EBPP implementation. RESULTS: One hundred local health departments participated (89% response rate) in the online survey. Most frequently implemented diabetes-related EBPPs in local health departments included: nutrition education for agency or community members; increased fruit and vegetable access in community settings; and community-wide campaigns to promote physical activity. Increased encouragement to others in the department to use evidence-based decision making and agency incentives to help employees use evidence-based decision making were positively associated with implementation of diabetes prevention and control EBPPs. CONCLUSIONS: Local health departments are the “front line” of public health and this study demonstrates the important role these organizations play in implementing diabetes prevention and control EBPPs. Potential leverage points for more widespread adoption of diabetes-related EBPPs in local health departments include education about and encouragement of evidence-based decision making and organizational incentives for employees to integrate evidence-based decision making into their diabetes prevention and control activities

Assessing tracheal health using optical metabolic imaging and optical coherence tomography

The health and pathophysiology of the tracheal mucosa is an important yet poorly understood aspect of respiratory medicine. Cilia, hair-like organelles important for mucociliary clearance, line the tracheal mucosa. Ciliary dysfunction leads to severe diseases such as primary ciliary dyskinesia and cystic fibrosis. Optical imaging can monitor ciliary function in vivo, ex vivo, and in vitro to understand the genesis of ciliary disease and potential treatment targets. Specifically, optical coherence tomography (OCT) has been used to quantify multiple parameters of ciliary motility in 2D and 3D. However, OCT inherently lacks information about the biochemical or metabolic state of cells. Optical metabolic imaging (OMI) quantitatively assesses cellular metabolism by imaging the autofluorescence intensities of endogenous metabolic co-enzymes nicotinamide dinucleotide (NADH) and flavin adenine dinucleotide (FAD). OMI probes the optical redox ratio (NADH intensity divided by FAD intensity), which is sensitive to the relative amounts of electron donors and acceptors within a cell. Ciliary function is highly dependent on ATP and therefore tightly linked to NADH and FAD levels through multiple metabolic pathways Please click Additional Files below to see the full abstract

Single cell metabolic imaging of tumor and immune cells in vivo in melanoma bearing mice

IntroductionMetabolic reprogramming of cancer and immune cells occurs during tumorigenesis and has a significant impact on cancer progression. Unfortunately, current techniques to measure tumor and immune cell metabolism require sample destruction and/or cell isolations that remove the spatial context. Two-photon fluorescence lifetime imaging microscopy (FLIM) of the autofluorescent metabolic coenzymes nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD) provides in vivo images of cell metabolism at a single cell level.MethodsHere, we report an immunocompetent mCherry reporter mouse model for immune cells that express CD4 either during differentiation or CD4 and/or CD8 in their mature state and perform in vivo imaging of immune and cancer cells within a syngeneic B78 melanoma model. We also report an algorithm for single cell segmentation of mCherry-expressing immune cells within in vivo images.ResultsWe found that immune cells within B78 tumors exhibited decreased FAD mean lifetime and an increased proportion of bound FAD compared to immune cells within spleens. Tumor infiltrating immune cell size also increased compared to immune cells from spleens. These changes are consistent with a shift towards increased activation and proliferation in tumor infiltrating immune cells compared to immune cells from spleens. Tumor infiltrating immune cells exhibited increased FAD mean lifetime and increased protein-bound FAD lifetime compared to B78 tumor cells within the same tumor. Single cell metabolic heterogeneity was observed in both immune and tumor cells in vivo.DiscussionThis approach can be used to monitor single cell metabolic heterogeneity in tumor cells and immune cells to study promising treatments for cancer in the native in vivo context

Autofluorescence lifetime imaging to monitor immune cell metabolism and function

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Dysfunctional BMPR2 signaling drives an abnormal endothelial requirement for glutamine in pulmonary arterial hypertension

Pulmonary arterial hypertension (PAH) is increasingly recognized as a systemic disease driven by alteration in the normal functioning of multiple metabolic pathways affecting all of the major carbon substrates, including amino acids. We found that human pulmonary hypertension patients (WHO Group I, PAH) exhibit systemic and pulmonary-specific alterations in glutamine metabolism, with the diseased pulmonary vasculature taking up significantly more glutamine than that of controls. Using cell culture models and transgenic mice expressing PAH-causing BMPR2 mutations, we found that the pulmonary endothelium in PAH shunts significantly more glutamine carbon into the tricarboxylic acid (TCA) cycle than wild-type endothelium. Increased glutamine metabolism through the TCA cycle is required by the endothelium in PAH to survive, to sustain normal energetics, and to manifest the hyperproliferative phenotype characteristic of disease. The strict requirement for glutamine is driven by loss of sirtuin-3 (SIRT3) activity through covalent modification by reactive products of lipid peroxidation. Using 2-hydroxybenzylamine, a scavenger of reactive lipid peroxidation products, we were able to preserve SIRT3 function, to normalize glutamine metabolism, and to prevent the development of PAH in BMPR2 mutant mice. In PAH, targeting glutamine metabolism and the mechanisms that underlie glutamine-driven metabolic reprogramming represent a viable novel avenue for the development of potentially disease-modifying therapeutics that could be rapidly translated to human studies

Metabolic Imaging of Head and Neck Cancer Organoids

<div><p>Head and neck cancer patients suffer from toxicities, morbidities, and mortalities, and these ailments could be minimized through improved therapies. Drug discovery is a long, expensive, and complex process, so optimized assays can improve the success rate of drug candidates. This study applies optical imaging of cell metabolism to three-dimensional <i>in vitro</i> cultures of head and neck cancer grown from primary tumor tissue (organoids). This technique is advantageous because it measures cell metabolism using intrinsic fluorescence from NAD(P)H and FAD on a single cell level for a three-dimensional <i>in vitro</i> model. Head and neck cancer organoids are characterized alone and after treatment with standard therapies, including an antibody therapy, a chemotherapy, and combination therapy. Additionally, organoid cellular heterogeneity is analyzed quantitatively and qualitatively. Gold standard measures of treatment response, including cell proliferation, cell death, and <i>in vivo</i> tumor volume, validate therapeutic efficacy for each treatment group in a parallel study. Results indicate that optical metabolic imaging is sensitive to therapeutic response in organoids after 1 day of treatment (p<0.05) and resolves cell subpopulations with distinct metabolic phenotypes. Ultimately, this platform could provide a sensitive high-throughput assay to streamline the drug discovery process for head and neck cancer.</p></div

Autofluorescence images show the redox ratio and fluorescence lifetimes of NAD(P)H and FAD in head and neck cancer organoids treated for 1 day with cetuximab, cisplatin, or their combination.

<p>NAD(P)H and FAD autofluorescence images were acquired from the same fields of view, and the redox ratio (top row), NAD(P)H fluorescence lifetime (middle row), and FAD fluorescence lifetime (bottom row) were calculated. For the redox ratio and fluorescence lifetimes, blue represents a low value and yellow represents a high value (see colorbars). Scale bar = 50um.</p

Untreated organoids contain cells with high levels of NAD(P)H intensity and cells with low levels of NAD(P)H intensity.

<p>(A) A representative fluorescence intensity image shows organoids with two levels of NAD(P)H intensity. (B) Low NAD(P)H cells exhibit a lower optical redox ratio than high NAD(P)H cells. (C) Low and High NAD(P)H cells exhibit similar NAD(P)H lifetimes. (D) Low NAD(P)H cells exhibit a higher FAD lifetime than high NAD(P)H cells. Scale bar = 50um. *p<0.05, t-test, n~50–100 cells per group.</p