Number, Density, and Surface/Cytoplasmic Distribution of GABA Transporters at Presynaptic Structures of Knock-In Mice Carrying GABA Transporter Subtype 1–Green Fluorescent Protein Fusions

GABA transporter subtype 1 (GAT1) molecules were counted near GABAergic synapses, to a resolution of ∼0.5 μm. Fusions between GAT1 and green fluorescent protein (GFP) were tested in heterologous expression systems, and a construct was selected that shows function, expression level, and trafficking similar to that of wild-type (WT) GAT1. A strain of knock-in mice was constructed that expresses this mGAT1–GFP fusion in place of the WT GAT1 gene. The pattern of fluorescence in brain slices agreed with previous immunocytochemical observations. [^3H]GABA uptake, synaptic electrophysiology, and subcellular localization of the mGAT1–GFP construct were also compared with WT mice. Quantitative fluorescence microscopy was used to measure the density of mGAT1–GFP at presynaptic structures in CNS preparations from the knock-in mice. Fluorescence measurements were calibrated with transparent beads and gels that have known GFP densities. Surface biotinylation defined the fraction of transporters on the surface versus those in the nearby cytoplasm. The data show that the presynaptic boutons of GABAergic interneurons in cerebellum and hippocampus have a membrane density of 800–1300 GAT1 molecules per square micrometer, and the axons that connect boutons have a linear density of 640 GAT1 molecules per micrometer. A cerebellar basket cell bouton, a pinceau surrounding a Purkinje cell axon, and a cortical chandelier cell cartridge carry 9000, 7.8 million, and 430,000 GAT1 molecules, respectively; 61–63% of these molecules are on the surface membrane. In cultures from hippocampus, the set of fluorescent cells equals the set of GABAergic interneurons. Knock-in mice carrying GFP fusions of membrane proteins provide quantitative data required for understanding the details of synaptic transmission in living neurons

A preliminary assessment of the efficiency of using drones in land cover mapping

This study represents a preliminary assessment of the efficiency of drones in surveying land cover at both large (c: 10 ha) and smaller (1m2) spatial scales. A DJI Mavic 2 drone was used to image the entire area of study and an orthomosaic was produced. This was converted into a land cover map through k-means clustering, with k = 3, where `Vegetation', `Bedrock' and `Bare soil' corresponded to the land cover categories. Regions of interest (ROIs) were selected and sub- sequently surveyed from close range. The correspondence between predicted land cover (pLC) and observed land cover (oLC) was then assessed. On a large spatial scale, absolute correspondence was present between pLC and oLC. In terms of relative representation of land cover categories, `Vegetation' was the only significantly correlated category across pLC and oLC, whilst the analogous correlations for `Bedrock' and `Bare soil' were weaker. The lower correspondence between pLC and oLC for `Bedrock' and `Bare soil' was due to the low value of k = 3 in the k-means clustering algorithm. This constrains a mixture of land covers into just one land cover category, with consequent reduction of the correlation between pLC and oLC. The method's accuracy and cost-effectiveness were compared to that of standard methods for land cover surveying. The entire process, including verification and orthomosaic land cover map processing times, approximated 32 hours. Consequently, this method is much shorter than standard surveys, which take days or weeks, and also requires less manpower.peer-reviewe

Emotion Regulation Goals Influence Strategy Use and Outcomes

Functionalist theories of emotion posit that people regulate their emotions in ways that help them accomplish their goals, suggesting that goals may be important for strategy selection. Two studies were conducted to examine reappraisal and suppression use when pursuing emotional and instrumental goals, and to assess the utility of those strategies in achieving distinct goals. Both studies found a stronger link between emotional goals and reappraisal than between emotional goals and suppression, but found no preference between strategies when pursuing an instrumental goal. Study 1 found that reappraisal had higher utility than suppression in achieving emotional goals, but not instrumental goals. In Study 2, individuals who used suppression more experienced more negative emotion and thought they made a worse impression on their partner, but they were not actually seen more negatively by others. Together these studies suggest that emotional goals may influence strategy selection and that strategies differ in their utility

Characterization and Function of Islet Antigen Presenting Cells during NOD Diabetes

Here we characterized the initial antigen presenting cells (APCs) within the islet of Langerhans to ascertain their identity and functional role as it pertains to autoimmune diabetes. The activation of the adaptive immune system is induced by the innate immune system, and more specifically APCs. Therefore, it is crucial to identify the APCs that are initiating T1D in order to elucidate the break in tolerance and intervene in order to inhibit progression. We have found that there is a resident macrophage that is present in all strains of mice. This islet macrophage has a distinct transcriptional profile that is unique when compared to other non-barrier tissue resident macrophages. The islet resident macrophage’s phenotype is akin to those macrophages found at barrier sites, i.e. the lung and the intestines. The barrier macrophages are constantly in contact with environmental pathogens, but the islet resident macrophage is located in tightly clustered mini-organs that are not in contact with barrier surfaces. We were able to show by RNAseq analysis that the islet resident macrophage is similar to macrophages treated with LPS and, thus, highly inflammatory. Furthermore, transcripts for the inflammatory cytokines TNFα and IL-1β found in islet macrophages were abundant and also were being produced in high amounts as protein. However, we were unable to definitively ascertain any functional role these cytokines have whether that may be inflammatory or homeostatic. Finally, the islet macrophages found in NOD.Rag1-/- mice were homogenous, i.e. single cell qRT-PCR displayed similar gene transcripts being expressed across all cells examined. However, although ~75% of the NOD islet macrophages expressed similar transcripts as the NOD.Rag1-/- macrophages, the remaining macrophages expressed an increase in inflammatory transcripts that are associated with interferon signaling. Finally, when compared to the non-diabetic B6.g7 islet macrophage, the NOD macrophage expressed differing chemokines that could be involved in chemotaxis of autoimmune cells into the islets. In non-diabetic mouse strains, the only leukocyte found within the islets is the resident macrophage. In the NOD mouse, however, at approximately the age of weaning, a CD103+ dendritic cell (DC) is found within the islets. This DC population enters at a similar time that CD3ε+ T cells are entering, however, we find that the initial entry of the CD103+ DC is not dependent on CD3ε+ T cell entry because the CD103+ DCs represent a small fraction of the myeloid compartment even in NOD.Rag1-/- mice. As diabetes progresses, these CD103+ DCs increase and this increase is dependent upon autoreactive CD3ε+ T cell entry. By genetic deletion of the CD103+ DC subset of DCs (cDC1 DCs) by utilizing the Batf3-deficient mouse, diabetes was abolished. The cDC1 DCs were not present in the islets or in any tissue examined, as expected. The protection from diabetes produced by the loss of the cDC1 DCs was absolute. At no timepoint was there infiltration of autoreactive cells to the islet of Langerhans assayed by histology and flow cytometry. The CD3ε+ T cell infiltration never exceeded that of baseline seen in non-diabetic strains. The islet gene expression profile of the NOD.Batf3-/- mouse was essentially identical to the lymphocyte deficient NOD.Rag1-/- islets. The priming of autoreactive CD8+ T cells was extinguished and priming of autoreactive CD4+ T cells was reduced by half. Transfer of naïve autoreactive T cells did not illicit entry into the islets or diabetes. However, diabetic splenocytes were able to confer diabetes when transferred into NOD.Batf3-/- mice. Therefore, the data strongly suggests the block in progression witnessed in the cDC1 DC deficient mouse is a lack of autoreactive T cell priming. In conclusion, we have identified the major APCs residing within the islet of Langerhans in the NOD mouse. During the life of the mouse, there is found a resident macrophage resting in a basal inflammatory state. Upon weaning, a CD103+ DC enters the islet of Langerhans but only in the NOD strain. The NOD islet macrophage expresses aberrant chemokines when compared to non-diabetic strains, which may lead to the initial infiltration of either the CD3ε+ T cells or the CD103+ DCs. When the CD103+ DCs are absent, T1D is halted due to a block in T cell priming. Although the definitive trigger of diabetes is yet to be determined, we believe that within the interplay amongst the islet resident macrophage, the entering T cells, and CD103+ DC is the initiating events. Perturbing one, or all, of the arms of this triad will result in a block in diabetes. How they interact, activate, and propagate the process will serve as the basis for future studies within the lab

Eduardo Araujo-Trujillo v. Attorney General United States

Agenc

Bodies of Parchment: Representing the Passion and Reading Manuscripts in Late Medieval England

In a diverse range of late-fourteenth- and fifteenth-century devotional literature, Christ\u27s body is metaphorically related to a book or a document at the moment of his crucifixion. His skin transforms into parchment, whips and scourges become pens, and a steady flow of blood, of ink, covers his body and the written page. And each word written onto his parchment body welcomes sustained study, acting as a potential meditative focal point for the devout reader. Through this metaphor and the accompanying materiality of the texts that include it, medieval authors and audiences could imagine intimately interacting with Christ\u27s body during the violence of his Passion. They could touch it, see it, hear it as it was read aloud, and, in the case of scribes, write it. This dissertation explores how the object of the text allowed audiences to participate actively in the events of Christ\u27s Passion and considers how the affective engagement with Christ\u27s suffering body that pervaded late-medieval devotional practice informed, in turn, the signifying power of a text\u27s materiality. The Christ-as-book metaphor often occurs in works that represent the creation and consumption of books in similar terms — terms that emphasize books not as already completed objects but as objects that are continuously in the process of being made, reproduced, edited, and circulated. Manuscript books are shown to function as loci for active and varied acts of interpretation as readers approached them as textual and material, but also visual and aural, objects. While the Christ-as-book metaphor can illuminate the dynamic role manuscripts could play in inspiring affective devotion, it has primarily been examined for the insight it offers into literary or theological trends of mysticism and popular religion. I argue, however, that this metaphor and the texts containing it can be read productively through the lenses of manuscript studies and book history. Building upon the work of D. F. McKenzie, Roger Chartier, and Alexandra Gillespie, I consider how the material form in which these texts occur influenced their reception and status as cultural objects amongst late-medieval lay audiences. The materiality of the manuscript book, I suggest, powerfully guided the ways in which readers approached, viewed, and experienced devotional works and, as a result, Christ\u27s body in late medieval England. The first chapter presents an examination of how writing, as the literal act of inscription and as a metaphor for the reading process: one remembered what one read, for example, by writing it in one\u27s heart), works to reproduce Christ\u27s suffering body both on the page and within the reader. By highlighting the interactions between pain and reading and between Christ\u27s body and a text\u27s material form, the Meditations on the Life and Passion of Christ, the Orison of the Passion, and the Charters of Christ illustrate that each re-reading of their respective texts constituted a re-writing as well. Every inscription of the text and of the events of Christ\u27s Passion produces a new copy — a new witness — both on the manuscript page and within the reader\u27s heart. Christ\u27s suffering body is not presented in these works as a reified, stable text to be copied out, or read, passively but instead as a text that welcomed readerly and scribal interpretation and reinvention. In the second chapter, I consider the potential for the material object of the text to function as a type of affective image for the devout reader. I argue that the ABC of Christ\u27s Passion, a text that relates the letters of the alphabet to the wounds inflicted upon Christ\u27s body, demonstrates how even the undecorated manuscript page could function as a potent “image” and focal point for a devout reader\u27s meditative practice. John Lydgate\u27s Passion poetry further illustrates the value of contemplating the visual aspects of a written text and its material form. While a reader could be moved to devout thoughts by regarding an image of the crucified Christ, Lydgate explores the potential for the object of the text to interact with and at times even supplant the devotional influence of standard Passion iconography. The layout of books and rolls can provide further evidence of how these textual objects were read by medieval audiences, and, in the third chapter, I investigate how affective reading practices mirrored the late-medieval devotional preoccupation with Christ\u27s suffering, fragmented body. The format and narrative structure of the Symbols of the Passion encourages readers to consult its text in a discontinuous fashion, as if the reader\u27s engagement with that work were intended to mirror how he or she would have meditated upon the partitioned body of Christ represented in popular arma Christi images. A parity existed, I suggest, between how Christ\u27s body and texts could be approached; both could be productively read in parts, with each fragment operating as an affectively-potent whole in its own right. Rather than being an idiosyncratic reading style promoted by the Symbols, the Book of Margery Kempe, Handlyng Synne, and a wide-range of other late-fourteenth and early-fifteenth century works of devotion also encourage late-medieval audiences to read discontinuously, indicating that this method of reading was influential and pervasive amongst a range of late-medieval devout audiences. But many audiences “read” medieval works by listening to them being read aloud rather than by personally and privately consulting a text within its manuscript context. In the fourth chapter, I study a unique musical version of the Short Charter of Christ and a non-musical: but nonetheless melodic) O-and-I lyric, “Throw hys hond.” I examine how these works explore the tension between spoken and written language and suggest that they provide a glimpse of the oral / aural potential latent within the metaphor of Christ\u27s body as a book. While manuscripts might be seen today as offering a silent material witness to the past, medieval audiences conceived of these forms as being imbued with sound, the otherwise dead skin and ink infused with a vocal presence

Six Key Elements of High Quality Technical Writing

What is the definition of “high quality”, when it comes to technical writing? What are the key elements that differentiate high-quality technical writing from the technical writing that is less effective in communicating the message? This webinar stems from the presenter’s observations and personal thoughts over the last decade or two, as an active researcher, a mentor, and a teacher in the engineering community. This webinar will be organized around six key elements or keywords identified by the presenter, i.e., innovation, engaging, hypothesis, logic, synthesis, and details

Coupling of Oxidation-Reduction Reactions of Chromium, Iron and Manganese: Implications for the Fate and Mobility of Chromium in Aquatic Environments

Both within the United States and internationally, hexavalent chromium (Cr(VI)) is a contaminant of concern in drinking water supplies. The U.S. Environmental Protection Agency is considering a Cr(VI)-specific standard. Thus improved technologies for Cr(VI) removal in drinking water are needed. Iron electrocoagulation for Cr(VI) removal was examined at conditions directly relevant to drinking water treatment, and humic acid (HA) affects the performance of electrocoagulation in multiple ways. The success of the chromium treatment or remediation also relies on the stability of the Cr(III)-containing solids with respect to reoxidation under groundwater conditions. Manganese is ubiquitous in aquatic and terrestrial environments, and the redox cycling of manganese may significantly impact the fate and transport of chromium. Coupling of redox reactions of chromium, iron and manganese involves multiple interaction pathways that occur in the aqueous phase as well as at solid-water interfaces. A mechanistic and quantitative understanding of these processes is needed to establish input parameters for kinetic and transport models and to enable decision-making for chromium treatment strategies. Iron electrocoagulation (EC) is a technology that can successfully achieve low concentrations of Cr(VI) in treated drinking water. In our research we have applied iron electrocoagulation (EC) with iron serving as the sacrificial anode to treat simulated drinking water solutions. Experiments have evaluated the effects of pH, dissolved oxygen, and common anions on Cr(VI) removal during batch EC treatment. In addition, the presence of humic acid (HA) inhibited the rate of Cr(VI) removal in electrocoagulation, with slower Cr(VI) removal at higher pH. This is due to dissolved oxygen competing with Cr(VI) for the oxidation of Fe(II) released from the anode. As determined using dynamic light scattering and wet chemistry experiments, the presence of HA resulted in the formation of Cr(III)-Fe(III)-HA colloids during electrocoagulation, which is difficult to remove in following water treatment steps of sedimentation and granular media filtration. Characterization of the solids by X-ray diffraction indicates that the iron oxides produced are lepidocrocite at pH 8, with more ferrihydrite in the presence of HA. Building on previous knowledge of MnO2 as an oxidant for Cr-containing solids, we systematically evaluated the rates and products of the oxidation of Cr(III) in iron oxides by MnO2. We found that Cr(III) dissolution from CrxFe1-x(OH)3 greatly influenced the Cr(VI) production rates. A multi-chamber reactor was used to assess the role of solid-solid mixing in CrxFe1-x(OH)3-MnO2 interactions. A dialysis membrane divided the reactor into two chambers, eliminating the possibility of direct contact of the solids in each chamber but allowing dissolved species to diffuse across the membrane. The Cr(VI) production rate was much lower in multi-chamber experiments (CrxFe1-x(OH)3||MnO2) than in completely mixed batch experiments under the same condition, indicating that the redox interaction is greatly accelerated by mixing of the two solids. The model was first established to predict Cr(VI) release in Cr(OH)3||MnO2 multichamber experiments, as dissolved Cr(III) concentration in equilibrium with Cr(OH)3 is higher at low pH and it’s easy to observe the behavior of Cr(VI) dynamics with more Cr(VI) generation. While solid phase Mn(IV) is well known oxidants of Cr(III)-containing solids, the localized oxidation of adsorbed Mn(II) by dissolved oxygen can also promote the oxidation of Cr(III) contained within CrxFe1-x(OH)3. The promotional effects was likely due to Mn redox cycling in which oxidized forms of Mn species were generated as oxidants of CrxFe1-x(OH)3 that were more potent than O2

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xviii, 712 hlm.; 21.5 x 15 c

WHY USE PRACTICAL TASKS AND LEARNING EXPERIENCES

A review of current and past theories of cognitive development focusing on the role of experiential learning is linked to implications for practice as well as to current Latvian policies and national directions. As can be seen, learning through practice is made up of several phases of experiences which impact on the student’s with disabilities motivation, skills and attitudes which lead to new learning. As well the subject, Home Economics and its Technologies is used as an example of an experiential learning structure which utilises previous experiences to enhance current and future learnin