Past research has shown that the processing of emotional visual stimuli and visual attention are tightly linked together. In particular, emotional stimuli processing can modulate attention, and, reciprocally, the processing of emotional stimuli can be facilitated or inhibited by attentional processes. However, our understanding of these interactions is still limited, with much work remaining to be done to understand the characteristics of this reciprocal interaction and the different mechanisms that are at play. This thesis presents a series of experiments which use eye-tracking, behavioural and event-related potential (ERP) methods in order to better understand these interactions from a cognitive and neuroscientific point of view. First, the influence of emotional stimuli on eye movements, reflecting overt attention, was investigated. While it is known that the emotional gist of images attracts the eye (Calvo and Lang, 2004), little is known about the influence of emotional content on eye movements in more complex visual environments. Using eye-tracking methods, and by adapting a paradigm originally used to study the influence of semantic inconsistencies in scenes (Loftus and Mackworth, 1978), we found that participants spend more time fixating emotional than neutral targets embedded in visual scenes, but do not fixate them earlier. Emotional targets in scenes were therefore found to hold, but not to attract, the eye. This suggests that due to the complexity of the scenes and the limited processing resources available, the emotional information projected extra-foveally is not processed in such a way that it drives eye movements. Next, in order to better characterise the exogenous deployment of covert attention toward emotional stimuli, a sample of sub-clinically anxious individuals was studied. Anxiety is characterised by a reflexive attentional bias toward threatening stimuli. A dot-probe task (MacLeod et al., 1986) was designed to replicate and extend past findings of this attentional bias. In particular, the experiment was designed to test whether the bias was caused by faster reaction times to fear-congruent probes or slower reaction times to neutral-congruent probes. No attentional bias could be measured. A further analysis of the literature suggests that subliminal cue stimulus presentation, as used in our case, may not generate reliable attentional biases, unlike longer cue presentations. This would suggest that while emotional stimuli can be processed without awareness, further processing may be necessary to trigger reflexive attentional shifts in anxiety. Then the time-course of emotional stimulus processes and its modulation by attention was investigated. Modulations of the very early visual ERP C1 component by emotional stimuli (e.g. Pourtois et al., 2004; Stolarova et al., 2006), but also by visual attention (Kelly et al., 2008), were reported in the literature. A series of three experiments were performed, investigating the interactions between endogenous covert spatial attention and object-based attention with emotional stimuli processing in the C1 time window (50–100 ms). It was found that emotional stimuli modulated the C1 only when they were spatially attended and task-irrelevant. This suggests that whilst spatial attention gates emotional facial processing from the earliest stages, only incidental processing triggers a specific response before 100 ms. Additionally, the results suggest a very early modulation by feature-based attention which is independent from spatial attention. Finally, simulated and actual electroencephalographic data were used to show that modulations of early ERP and event-related field (ERF) components are highly dependent on the high-pass filter used in the pre-processing stage. A survey of the literature found that a large part of ERP/ERF reports (about 40%) use high-pass filters that may bias the results. More particularly, a large proportion of papers reporting very early modulations also use such filters. Consequently, a large part of the literature may need to be re-assessed. The work described in this thesis contributes to a better understanding of the links between emotional stimulus processing and attention at different levels. Using various experimental paradigms, this work confirms that emotional stimuli processing is not ‘automated’, but highly dependent on the focus of attention, even at the earlier stages of visual processing. Furthermore, the uncovered potential bias generated by filtering will help to improve the reliability and precision of research in the ERP/ERF field, and more particularly in studies looking at early effects.

Nicotinamide delivered in drinking water at about 2 g/kg/day significantly prolonged survival and showed a suggestive improvement on memory in the Npc1 (nih) / Npc1 (nih) mouse model of infantile NPC1 disease. It is likely that this role is due to its function as a histone deacetylase (HDAC) inhibitor although another HDAC inhibitor, valproic acid, was without effect. Nicotinamide could also work by preventing/reversing oxidative stress.

UA Open Access Publishing Fund / Background: Atlantoaxial surgical fixation is widely employed treatment strategy for a myriad of pathologies affecting the stability of the atlantoaxial joint. The most common technique used in adults, and in certain cases in children, involves a posterior construct with C1 lateral mass screws, and C2 pars or pedicle screws. This technical note aims to provide a step‑by‑step guide to this procedure using cadaveric and fluoroscopic images. Methods: An embalmed, human, cadaveric, specimen was used for this study. The subject did not have obvious occipital‑cervical pathology. Dissections and techniques were performed to mimic actual surgical technique. Photographs were taken during each step, and the critical aspects of each step were highlighted. Fluoroscopic images from a real patient undergoing C1/C2 fixation were also utilized to further highlight the anatomic‑radiographic relationships. This study was performed without external or industry funding. Results: Photographic and radiographic pictures and drawings are presented to illustrate the pertinent anatomy and technical aspects of this technique. The nuances of each step, including complication avoidance strategies are also highlighted. Conclusions: Given the widespread utilization of this technique, described step‑by‑step guide is timely for surgeons and trainees alike.

Thesis advisor: Amir H. Hoveyda / Chapter 1: Efficient Z-Selective Cross-Metathesis of Secondary Allylic Ethers Efficient Z-selective cross-metathesis of secondary allylic ethers were catalyzed by monoaryloxide monopyrrolide molybdenum complexes. Reactions involving both silyl and benzyl protected ethers were demonstrated, as well as ethers containing alkyl, aryl and alkynyl substituents. Mechanistic studies were performed, and the reactions were applied to the total synthesis of several ene-diyne natural products. Chapter 2. Stereoselective Total Synthesis of Disorazole C1 The stereoselective total synthesis of disorazole C1 is reported. The synthesis was completed in 12 longest linear steps. Our synthesis demonstrates the utility of Z-selective cross-metathesis to form both alkenyl borons and alkenyl halides. Another key transformation was a one-pot Suzuki-dimerization reaction to form a symmetric 30 membered ring in relatively high yield. Chapter 3. Stereoselective Cross-Metathesis to Form Trisubstituted Alkenes Initial studies into the stereoselective formation of trisubstituted olefins through molybdenum catalyzed cross-metathesis have been performed. Our mechanistic understanding of the reaction lead us to focus on the synthesis of alkenyl halides, which can be obtained in up 90% yield and 75:25 E:Z selectivity. Chapter 4: Ring-Closing Metathesis in the Synthesis of Natural Products Development of highly efficient and selective ring-closing metathesis reactions have enabled collaborators to successfully implement routes in total synthesis endeavors. A diastereoselective seven-membered ring-closing metathesis enabled the successful synthesis of (±)-tetrapetalone A methyl-aglycon. An enantioselective ring-closing metathesis to form a six membered ring has provided access to enantioenriched aspidosperma alkaloids. / Thesis (PhD) — Boston College, 2016. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Chemistry.

Ebolavirus (EboV) is a highly pathogenic enveloped virus that causes outbreaks of zoonotic infection in Africa. The clinical symptoms are manifestations of the massive production of pro-inflammatory cytokines in response to infection and in many outbreaks, case fatality rate exceeds 75%. The unpredictable onset, ease of transmission, rapid progression of disease, high mortality and lack of effective vaccine or therapy have created a high level of public concern about EboV. Here we report the properties of a benzylpiperazine adamantane diamide-derived compound identified in a screen for inhibitors of EboV infection. We found that the inhibitor is specific, reversible, and that the target(s) for inhibition are present in cells and not in virus particles. The compound is not an inhibitor of acid pH-dependent endosome protease activity, which is required for EboV infection. Treatment of cells with this compound causes accumulation of cholesterol in late endosomes and lysosomes (LE/LY), suggesting it inhibits one or more proteins involved in regulation of cholesterol uptake into cells. Using mutant cell lines and informative derivatives of the inhibitor, we found the inhibitor target is the endosomal membrane protein Niemann-Pick C1 (NPC1). NPC1 is a polytopic LE/LY membrane protein that mediates uptake of lipoprotein-derived cholesterol into cells. We find that NPC1 is essential for EboV infection, that NPC1 binds to the protease-cleaved GP1 subunit of the EboV glycoprotein, and that the anti-viral compound inhibits infection by targeting NPC1 and interfering with binding to GP1. Furthermore, analysis of viral variants resistant to the anti-viral compound revealed that the residues which confer resistance are located on the surface of the receptor binding domain of GP1. Combined with the results of previous studies of GP structure and function, our findings support a model of EboV infection in which cleavage of the GP1 subunit by endosomal cathepsin proteases removes heavily glycosylated domains to expose the N-terminal domain, which is a ligand for NPC1 and regulates membrane fusion by the GP2 subunit. Thus, NPC1 is essential for EboV entry and a target for anti-viral therapy.

Reduced Hsieh-Clough-Tocher elements are triangular C1-elements with only nine degrees of freedom. Simple formulas for the basis functions of reduced Hsieh-Clough-Tocher elements based on the edge vectors of the triangle have been given recently for a barycentric splitting. We generalise these formulas to the case of an arbitrary splitting point.

This report is an investigation of regioselectivity in transition metal catalyzed ring-opening reactions involving unsymmetrical oxabicycles, specifically with a substituent at the C1 position. This report also provides the details of the work conducted towards the preparation of various oxanorbornadienes and their precursors. A large number of reactions have been developed using various transition metal catalysts on oxabicyclic alkenes to form functionalized organic scaffolds. However, most of the literature is limited to symmetrical substrates. Introduction of a substituent at the bridgehead carbon of the bicyclic ring makes the molecule unsymmetrical. The implications of loss of the plane of symmetry in C1 substituted oxabicyclic ring are manifested in interesting ways during various metal catalyzed reactions. The fundamental basis for the current work is to study the consequences of transition metal catalyzed ring opening reactions of unsymmetrical bicyclic alkenes. The reactivity of a wide range of C1 substituted benzoxanorbornadienes and oxanorbornadienes in palladium and nickel-catalyzed ring opening reactions was explored. The palladium catalyzed ring opening reaction of both electron rich and electron deficient C1 substituted benzoxanorbornadienes are optimized. The ring opening reactions with electron withdrawing C1 substituent resulted in formation of substituted naphthalene-1-carboxylic acid methyl ester derivatives in up to 85% yield. Electron donating substituents on the C1 position of benzoxanorbornadiene led to the formation of substituted cis-1,2-dihydronaphthol rings in excellent yields. Palladium catalyzed ring opening reactions were also explored with a wide range of aryl iodides and halobenzenes. The electronic and steric effects of the substituent at the C1 position of oxabicyles were also investigated. The nickel catalyzed ring opening reactions resulted in formation of inseparable regioisomeric mixtures of products. However, it was found that the nickel catalyzed ring opening of 1-methoxycarbonyl-7-oxabenzonorbornadiene occurred regioselectively affording a single product. A scalable procedure for preparation of large quantities of 2-bromofuran was developed. 2-Aryl furans were prepared using Suzuki cross coupling protocols of 2-bromofuran with aryl boronic acids whereas 2-alkyl furans were prepared by iron catalyzed cross coupling reaction of 2-bromofuran with various alkyl and cycloalkyl Grignard reagents. The 2-substituted furans were used for the preparation of novel C1 substituted benzoxanorbornadiene and oxanorbornadienes.

Each member of the protein kinase C (PKC) family activates cell signaling pathways with different and sometimes opposing cell functions, such as cell division, migration, or death. Because of the importance of these processes in human diseases and disorders like cancer, stroke, and Alzheimer’s disease, there is a need for drugs which modify the action of PKC. However, drug design is difficult due to the complicated nature of PKC regulation. To better understand the differential regulation of PKC activity, these studies probe the structure, dynamics, and reactivity of one of the domains responsible for PKC regulation, C1B. C1B binds signaling molecules and translocates PKC to membranes in order to release the kinase domain from inhibition. Mutagenesis and ligand-binding assays monitored with fluorescence and nuclear magnetic resonance (NMR) techniques show that a single variable residue in C1B dramatically affects the sensitivity to signal activators. Investigation of the domain structure and dynamics using NMR revealed the identity of this residue alters the dynamics of the activator binding loops, without changing the structure. NMR studies of the C1B variants in membrane-mimicking micelles showed this residue also changes the interaction of the regulatory domain with lipids. These results demonstrate PKC isoforms have evolved specific functions by tuning dynamics and membrane affinity. Alternatively, PKC can be activated by reactive oxygen species by a mechanism that does not require binding of signaling molecules or membrane localization. To investigate the role of C1B in this type of signaling, the regulatory domain reactivity is monitored via NMR and gel electrophoresis. These studies reveal a particular cysteine residue in C1B that is most reactive, an alternative conformation of C1B in which this residue is more exposed, and modification of C1B leads to unfolding and zinc loss. Because the regulatory domains are responsible for auto-inhibition of the kinase domain, C1B unfolding provides a plausible explanation for activation of PKC by reactive oxygen species. The relation of the intrinsic C1B properties to the activation of PKC can be used to develop drugs with a single mechanism and to better understand how closely related signaling proteins develop specific functions.

The physiological role of the plasma contact system still remains a partial enigma. The aim of the presented work was to expand our understanding of the plasma contact system, focusing on its physiological activation and function, principally from a hemostatic perspective. It also explored contact system activation under pathological conditions. We found that when human platelets become activated in blood, plasma proteins of the contact system bind to platelets and initiate contact activation. The platelet-triggered contact activation contributed to clot formation by shortening the clotting time and enhancing clot stability. We demonstrated that the regulation of contact activation elicited by activated platelets differed from the previously described contact activation elicited by negatively charged material surfaces. Platelet-triggered contact activation and activation propelled by clotting blood were found to be regulated by antithrombin, whereas material-induced activation was regulated by C1 inhibitor. We also showed that the fibrin fibers that are formed during the clot process further enhance and propagate the contact activation initially induced by activated platelets. Fibrin not only activated factor XII but also seemed to increase the affinity of antithrombin for the proteases of the contact system, leading to the generation of contact enzyme-antithrombin complexes during clot formation. To determine whether the contact system might be involved in the inflammation and vascular disease associated with systemic lupus erythematosus (SLE), we analyzed plasma samples from SLE patients. These patients were found to have altered levels of contact enzyme-serpin complexes, supporting the concept that the contact system may be involved in the pathophysiology of SLE. The contact enzyme-antithrombin complexes were clearly linked to platelet activation in vivo. Altered levels of both FXIIa-antithrombin and FXIIa-C1 inhibitor were found to be correlated with previous vascular disease and may therefore be potential biomarkers for assessing the risk of thrombotic events in SLE patients. These findings add to our knowledge of how the plasma contact system is activated and functions in vivo and will help us to understand the role of the contact system, not only in hemostasis but also in vascular disease and thrombotic conditions.

No description available.