Pharmacist-Physician Collaborative Practice to Improve Diabetes Care at Tampa General Medical Group.

Quality Improvement Success Stories are published by the American Diabetes Association in collaboration with the American College of Physicians and the National Diabetes Education Program. This series is intended to highlight best practices and strategies from programs and clinics that have successfully improved the quality of care for people with diabetes or related conditions. Each article in the series is reviewed and follows a standard format developed by the editors of Clinical Diabetes. The following article describes a pharmacist-physician collaborative effort to reduce A1C and blood pressure and thereby lower risks for complications for people with diabetes being treated at a network of family care clinics in the Tampa, FL, area.

Subthreshold glutamate release from mitral cell dendrites.

The dendrites of a number of neuron types function as presynaptic structures, releasing transmitter after action potentials and dendritic spikes. In this regard, dendrites can function like axons, producing discrete outputs after suprathreshold electrical events. However, as the major site of synaptic inputs, dendrites experience ongoing subthreshold fluctuations in membrane potential, raising the question of whether these subthreshold changes can cause changes in transmitter release. Here, we show that mitral cells of the accessory olfactory bulb release glutamate from their dendrites in response to both subthreshold and suprathreshold stimuli. Whereas subthreshold output was typically low under control conditions, it could be enhanced several fold by pharmacological or endogenous activation of group I metabotropic glutamate receptors. These results indicate that presynaptic dendrites can support two distinct forms of output, and can dynamically regulate how electrical activity is coupled to transmitter release.

A deep learning based method for large-scale classification, registration, and clustering of in-situ hybridization experiments in the mouse olfactory bulb.

BACKGROUND: The Allen Mouse Brain Atlas allows study of the brain's molecular anatomy at cellular scale, for thousands genes. To fully leverage this resource, one must register histological images of brain tissue - a task made challenging by the brain's structural complexity and heterogeneity, as well as inter-experiment variability. NEW METHOD: We have developed a deep-learning based methodology for classification and registration of thousands of sections of brain tissue, using the mouse olfactory bulb (OB) as a case study. RESULTS: We trained a convolutional neural network (CNN) to derive an image similarity measure for in-situ hybridization experiments, and embedded these in a low-dimensional feature space to guide the design of registration templates. We then compiled a high quality, registered atlas of gene expression for the OB (the first such atlas for the OB, to our knowledge). As proof-of-principle, the atlas was clustered using non-negative matrix factorization to reveal canonical expression motifs, and to identify novel, lamina-specific marker genes. COMPARISON WITH EXISTING METHODS: Our method leverages virtues of CNNs for a set of important problems in molecular neuroanatomy, with performance comparable to existing methods. CONCLUSION: The atlas we have complied allows for intra- and inter-laminar comparisons of gene expression patterns in the OB across thousands of genes, as well identification of canonical expression profiles through clustering. We anticipate that this will be a useful resource for investigators studying the bulb's development and functional topography. Our methods are publicly available for those interested in extending them to other brain areas.

Recurrent dendrodendritic inhibition of accessory olfactory bulb mitral cells requires activation of group I metabotropic glutamate receptors.

Metabotropic glutamate receptors (mGluRs) modulate neural excitability and network tone in many brain regions. Expression of mGluRs is particularly high in the accessory olfactory bulb (AOB), a CNS structure critical for detecting chemicals that identify kin and conspecifics. Because of its relative simplicity and its direct projection to the hypothalamus, the AOB provides a model system for studying how mGluRs affect the flow of encoded sensory information to downstream areas. We investigated the role of group I mGluRs in synaptic processing in AOB slices and found that under control conditions, recurrent inhibition of principal neurons (mitral cells) was completely eliminated by the mGluR1 antagonist LY367385 [(S)-(+)-alpha-amino-4-carboxy-2 methylbenzeneacetic acid]. In addition, the group I mGluR agonist DHPG [(S)-3,5-dihydroxyphenylglycine; 20 microM] induced a dramatic increase in the rate of spontaneous IPSCs. This increase was dependent on voltage-gated calcium channels but persisted even after blockade of ionotropic glutamatergic transmission and sodium channels. Together, these results indicate that mGluR1 plays a critical role in controlling information flow through the AOB and suggest that mGluR1 may be an important locus for experience-dependent changes in synaptic function.

Genome-scale investigation of olfactory system spatial heterogeneity.

The early olfactory system is organized in parallel, with numerous, specialized subsystems established by the modular and topographic projections of sensory inputs. While these anatomical sub-systems are in many cases demarcated by well-known marker genes, we stand to learn considerably more about their possible functional specializations from comprehensive, genome-scale descriptions of their molecular anatomy. Here, we leverage the resources of the Allen Brain Atlas (ABA)-a spatially registered compendium of gene expression for the mouse brain-to investigate the early olfactory system's genomic anatomy. We cluster thousands of genes across thousands of voxels in the ABA to derive several novel parcellations of the olfactory system, and concomitantly discover novel sets of enriched, subregion-specific genes that can serve as a starting point for future inquiry.

Tuft calcium spikes in accessory olfactory bulb mitral cells.

The mammalian accessory olfactory system is critical for the detection and identification of pheromones and the representation of complex stimuli including sex, genetic relatedness, and individual identity. Mitral cells, the principal cells of the accessory olfactory bulb (AOB), receive monosynaptic input from the sensory periphery and already show highly specific response properties, firing selectively for combinations of genetic markers and gender-specific cues. Vomeronasal sensory neuron axons form synapses onto distal tuft-like branches of mitral cell primary dendrites. We have studied dendritic excitability and synaptic integration in AOB mitral cell dendrites, and we show that dendrites of accessory olfactory bulb mitral cells support action potential propagation and can fire regenerative spike-like events that are likely to contribute to the integration of inputs to these cells. These tuft spikes may be important for the specificity of AOB mitral cell responses.

Reversal of cisplatin resistance with a BH3 mimetic, (-)-gossypol, in head and neck cancer cells: role of wild-type p53 and Bcl-xL.

Organ preservation protocols in head and neck squamous cell carcinoma (HNSCC) are limited by tumors that fail to respond. We observed that larynx preservation and response to chemotherapy is significantly associated with p53 overexpression, and that most HNSCC cell lines with mutant p53 are more sensitive to cisplatin than those with wild-type p53. To investigate cisplatin resistance, we studied two HNSCC cell lines, UM-SCC-5 and UM-SCC-10B, and two resistant sublines developed by cultivation in gradually increasing concentrations of cisplatin. The cisplatin-selected cell lines, UM-SCC-5PT and UM-SCC-10BPT, are 8 and 1.5 times more resistant to cisplatin than the respective parental cell lines, respectively. The parental lines overexpress p53 and contain p53 mutations but the cisplatin-resistant cell lines do not, indicating that cells containing mutant p53 were eliminated during selection. Bcl-x(L) expression increased in the cisplatin-resistant lines relative to the parental lines, whereas Bcl-2 expression was high in the parental lines and decreased in the cisplatin-resistant lines. Thus, cisplatin selected for wild-type p53 and high Bcl-x(L) expression in these cells. We tested a small-molecule BH3 mimetic, (-)-gossypol, which binds to the BH3 domain of Bcl-2 and Bcl-x(L), for activity against the parental and cisplatin-resistant cell lines. At physiologically attainable levels, (-)-gossypol induces apoptosis in 70% to 80% of the cisplatin-resistant cells but only in 25% to 40% of the parental cells. Thus, cisplatin-resistant cells seem to depend on wild-type p53 and Bcl-x(L) for survival and BH3 mimetic agents, such as (-)-gossypol, may be useful adjuncts to overcome cisplatin resistance in HNSCC.

Neuro at the Nanoscale: Diffraction-Unlimited Imaging with STED Nanoscopy.

Recent breakthroughs in fluorescence microscopy have pushed spatial resolution well beyond the classical limit imposed by diffraction. As a result, the field of nanoscopy has emerged, and diffraction-unlimited resolution is becoming increasingly common in biomedical imaging applications. In this review, we recap the principles behind STED nanoscopy that allow imaging beyond the diffraction limit, and highlight both historical and recent advances made in the field of neuroscience as a result of this technology.

The number of olfactory stimuli that humans can discriminate is still unknown.

It was recently proposed (Bushdid et al., 2014) that humans can discriminate between at least a trillion olfactory stimuli. Here we show that this claim is the result of a fragile estimation framework capable of producing nearly any result from the reported data, including values tens of orders of magnitude larger or smaller than the one originally reported in (Bushdid et al., 2014). Additionally, the formula used to derive this estimate is well-known to provide an upper bound, not a lower bound as reported. That is to say, the actual claim supported by the calculation is in fact that humans can discriminate at most one trillion olfactory stimuli. We conclude that there is no evidence for the original claim.

Olfaction, valuation, and action: reorienting perception.

In the philosophy of perception, olfaction is the perennial problem child, presenting a range of difficulties to those seeking to define its proper referents, and its phenomenological content. Here, we argue that many of these difficulties can be resolved by recognizing the object-like representation of odors in the brain, and by postulating that the basic objects of olfaction are best defined by their biological value to the organism, rather than physicochemical dimensions of stimuli. Building on this organism-centered account, we speculate that the phenomenological space of olfaction is organized into a number of coarse affective dimensions that apply categorically. This organization may be especially useful for coupling sensation to decision making and instrumental action in a sensory modality where the stimulus space is especially complex and high dimensional.

Categorical dimensions of human odor descriptor space revealed by non-negative matrix factorization.

In contrast to most other sensory modalities, the basic perceptual dimensions of olfaction remain unclear. Here, we use non-negative matrix factorization (NMF)--a dimensionality reduction technique--to uncover structure in a panel of odor profiles, with each odor defined as a point in multi-dimensional descriptor space. The properties of NMF are favorable for the analysis of such lexical and perceptual data, and lead to a high-dimensional account of odor space. We further provide evidence that odor dimensions apply categorically. That is, odor space is not occupied homogenously, but rather in a discrete and intrinsically clustered manner. We discuss the potential implications of these results for the neural coding of odors, as well as for developing classifiers on larger datasets that may be useful for predicting perceptual qualities from chemical structures.

Functional polarity in neurons: what can we learn from studying an exception?

Dendrites and axons typically handle very different aspects of neuronal signaling. However, many of the functional distinctions between these two types of processes are absent in neurons with release-competent dendrites. This raises fundamental questions about the molecular mechanisms that promote and permit functional specialization, and suggests that the 'exceptional' case of presynaptic dendrites may provide important clues on how neuronal polarity is established. To help stimulate thinking on this new front, we summarize some key aspects of the physiology of dendritic neurotransmitter release, together with recent work on the molecular basis of neuronal polarity.

Tissue-preserving approach to extracting DNA from paraffin-embedded specimens using tissue microarray technology.

BACKGROUND: DNA extracted from tumor cells or normal cells contained in formalin-fixed, paraffin-embedded tissues is widely used in many laboratories. The 2 most common procedures to isolate cells for DNA extraction from paraffin-embedded tissues are scalpel microdissection and laser capture microdissection. A new tissue- and time-conserving method for rapid DNA isolation from small cores taken from paraffin-embedded tissue blocks is described in this report. METHODS: DNA was extracted from small tissue cores collected from paraffin-embedded tissue blocks at the time of tissue microarray construction. The quality and quantity of the DNA extracted was compared to DNA collected by scalpel microdissection. DNA collected from tissue cores was used in polymerase chain reaction (PCR) and loss of heterozygosity (LOH) analysis. RESULTS: The quality and quantity of DNA obtained using tissue cores was comparable to DNA obtained by traditional methods. The tissue core method of DNA extraction preserves the tissue blocks from which the cores are extracted for future use. Adequate quantities of DNA can be successfully extracted from small segments of tissue cores and used for PCR. DNA isolated by tissue microdissection and the tissue core method were comparable when used to assess allelic heterozygosity on chromosome arm 18q. CONCLUSION: The tissue core method of DNA isolation is reliable, tissue conserving, and time effective. Tissue cores for DNA extraction can be harvested at the same time as tissue microarray construction. The technique has the advantage of preserving the original tissue blocks for additional study as only tiny cores are removed.

The relative detectability for mice of gaps having different ramp durations at their onset and offset boundaries.

The effect on gap detectability of varying noise fall time (FT) and rise time (RT) of the gap boundary ramps was examined in mice using reflex modification audiometry, measuring inhibition of acoustic startle reflexes by variously shaped gaps just preceding reflex expression. In experiment 1 (n = 12) inhibition increased up to near-asymptotic values with longer FT (0, 1, 2, 3, 5, or 10 ms) and QT (quiet time, 0 to 13 ms), with a 2:1 trade-off between FT and QT. In experiment 2 (n = 24) inhibition increased for any RT above 0 ms (2, 3, 5, or 7 ms) if QT= 1 ms, but diminished with increased RT when QT = 3 or 8 ms. Enhanced detectability for subthreshold gaps by longer ramps results from their extending the apparent gap duration. The negative effect of increased RT for threshold gaps suggests the importance for gap detection of the stronger neural responses to sharp edges at the end of the gap shown previously in the mouse inferior colliculus. These effects are specific to gaps: inhibition for fixed (70-dB SPL) or varied level pulses (30 to 60 dB) was unaffected by varying the ramped edges (experiments 3 and 4, n = 9).