Solid state NMR studies of structure and dynamics in systems of biological interest

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Chemistry, 1997.Includes bibliographical references.by Joanna R. Long.Ph.D

The interaction and orientation of Peptide KL4 in model membranes

We report on the orientation and location of synthetic pulmonary surfactant peptide KL4, (KLLLL)4K, in model lipid membranes. The partitioning depths of selectively deuterated leucine residues within KL4 were determined in DPPC:POPG (4:1) and POPC:POPG (4:1) bilayers by oriented neutron diffraction. These measurements were combined with an NMR-generated model of the peptide structure to determine the orientation and partitioning of the peptide at the lipid–water interface. The results demonstrate KL4 adopting an orientation that interacts with a single membrane leaflet. These observations are consistent with past 2H NMR and EPR studies (Antharam et al., 2009; Turner et al., 2014)

CLEAR I: Ages and Metallicities of Quiescent Galaxies at 1.0<z<1.8\mathbf{1.0 < z < 1.8} Derived from Deep Hubble Space Telescope Grism Data

We use deep \textit{Hubble Space Telescope} spectroscopy to constrain the metallicities and (\editone{light-weighted}) ages of massive ($\log M_\ast/M_\odot\gtrsim10$) galaxies selected to have quiescent stellar populations at $1.0<z<1.8$. The data include 12--orbit depth coverage with the WFC3/G102 grism covering $\sim$ $8,000<\lambda<11,500$~\AA\, at a spectral resolution of $R\sim 210$ taken as part of the CANDELS Lyman-$\alpha$ Emission at Reionization (CLEAR) survey. At $1.0<z<1.8$, the spectra cover important stellar population features in the rest-frame optical. We simulate a suite of stellar population models at the grism resolution, fit these to the data for each galaxy, and derive posterior likelihood distributions for metallicity and age. We stack the posteriors for subgroups of galaxies in different redshift ranges that include different combinations of stellar absorption features. Our results give \editone{light-weighted ages of $t_{z \sim 1.1}= 3.2\pm 0.7$~Gyr, $t_{z \sim 1.2}= 2.2\pm 0.6$~Gyr, $t_{z\sim1.3}= 3.1\pm 0.6$~Gyr, and $t_{z\sim1.6}= 2.0 \pm 0.6$~Gyr, \editone{for galaxies at $z\sim 1.1$, 1.2, 1.3, and 1.6. This} implies that most of the massive quiescent galaxies at $168$\% of their stellar mass by a redshift of $z>2$}. The posteriors give metallicities of \editone{$Z_{z\sim1.1}=1.16 \pm 0.29$~$Z_\odot$, $Z_{z\sim1.2}=1.05 \pm 0.34$~$Z_\odot$, $Z_{z\sim1.3}=1.00 \pm 0.31$~$Z_\odot$, and $Z_{z\sim1.6}=0.95 \pm 0.39$~$Z_\odot$}. This is evidence that massive galaxies had enriched rapidly to approximately Solar metallicities as early as $z\sim3$.Comment: 32 pages, 23 figures, Resubmited to ApJ after revisions in response to referee repor

Impaired natural killer cell phenotype and function in idiopathic and heritable pulmonary arterial hypertension

BACKGROUND: Beyond their role as innate immune effectors, natural killer (NK) cells are emerging as important regulators of angiogenesis and vascular remodeling. Pulmonary arterial hypertension (PAH) is characterized by severe pulmonary vascular remodeling and has long been associated with immune dysfunction. Despite this association, a role for NK cells in disease pathology has not yet been described. METHODS AND RESULTS: Analysis of whole blood lymphocytes and isolated NK cells from PAH patients revealed an expansion of the functionally defective CD56(-)/CD16(+) NK subset that was not observed in patients with chronic thromboembolic pulmonary hypertension. NK cells from PAH patients also displayed decreased levels of the activating receptor NKp46 and the killer immunoglobulin-like receptors 2DL1/S1 and 3DL1, reduced secretion of the cytokine macrophage inflammatory protein-1β, and a significant impairment in cytolytic function associated with decreased killer immunoglobulin-like receptor 3DL1 expression. Genotyping patients (n=222) and controls (n=191) for killer immunoglobulin-like receptor gene polymorphisms did not explain these observations. Rather, we show that NK cells from PAH patients exhibit increased responsiveness to transforming growth factor-β, which specifically downregulates disease-associated killer immunoglobulin-like receptors. NK cell number and cytotoxicity were similarly decreased in the monocrotaline rat and chronic hypoxia mouse models of PAH, accompanied by reduced production of interferon-γ in NK cells from hypoxic mice. NK cells from PAH patients also produced elevated quantities of matrix metalloproteinase 9, consistent with a capacity to influence vascular remodeling. CONCLUSIONS: Our work is the first to identify an impairment of NK cells in PAH and suggests a novel and substantive role for innate immunity in the pathobiology of this disease

Pain assessment for people with dementia: a systematic review of systematic reviews of pain assessment tools.

BACKGROUND: There is evidence of under-detection and poor management of pain in patients with dementia, in both long-term and acute care. Accurate assessment of pain in people with dementia is challenging and pain assessment tools have received considerable attention over the years, with an increasing number of tools made available. Systematic reviews on the evidence of their validity and utility mostly compare different sets of tools. This review of systematic reviews analyses and summarises evidence concerning the psychometric properties and clinical utility of pain assessment tools in adults with dementia or cognitive impairment. METHODS: We searched for systematic reviews of pain assessment tools providing evidence of reliability, validity and clinical utility. Two reviewers independently assessed each review and extracted data from them, with a third reviewer mediating when consensus was not reached. Analysis of the data was carried out collaboratively. The reviews were synthesised using a narrative synthesis approach. RESULTS: We retrieved 441 potentially eligible reviews, 23 met the criteria for inclusion and 8 provided data for extraction. Each review evaluated between 8 and 13 tools, in aggregate providing evidence on a total of 28 tools. The quality of the reviews varied and the reporting often lacked sufficient methodological detail for quality assessment. The 28 tools appear to have been studied in a variety of settings and with varied types of patients. The reviews identified several methodological limitations across the original studies. The lack of a 'gold standard' significantly hinders the evaluation of tools' validity. Most importantly, the samples were small providing limited evidence for use of any of the tools across settings or populations. CONCLUSIONS: There are a considerable number of pain assessment tools available for use with the elderly cognitive impaired population. However there is limited evidence about their reliability, validity and clinical utility. On the basis of this review no one tool can be recommended given the existing evidence

Relationships between the Circadian System and Alzheimer’s Disease-Like Symptoms in Drosophila

Circadian clocks coordinate physiological, neurological, and behavioral functions into circa 24 hour rhythms, and the molecular mechanisms underlying circadian clock oscillations are conserved from Drosophila to humans. Clock oscillations and clock-controlled rhythms are known to dampen during aging; additionally, genetic or environmental clock disruption leads to accelerated aging and increased susceptibility to age-related pathologies. Neurodegenerative diseases, such as Alzheimer’s disease (AD), are associated with a decay of circadian rhythms, but it is not clear whether circadian disruption accelerates neuronal and motor decline associated with these diseases. To address this question, we utilized transgenic Drosophila expressing various Amyloid-β (Aβ) peptides, which are prone to form aggregates characteristic of AD pathology in humans. We compared development of AD-like symptoms in adult flies expressing Aβ peptides in the wild type background and in flies with clocks disrupted via a null mutation in the clock gene period (per[superscript 01]). No significant differences were observed in longevity, climbing ability and brain neurodegeneration levels between control and clock-deficient flies, suggesting that loss of clock function does not exacerbate pathogenicity caused by human-derived Aβ peptides in flies. However, AD-like pathologies affected the circadian system in aging flies. We report that rest/activity rhythms were impaired in an age-dependent manner. Flies expressing the highly pathogenic arctic Aβ peptide showed a dramatic degradation of these rhythms in tune with their reduced longevity and impaired climbing ability. At the same time, the central pacemaker remained intact in these flies providing evidence that expression of Aβ peptides causes rhythm degradation downstream from the central clock mechanism

Loss of Extreme Long-Range Enhancers in Human Neural Crest Drives a Craniofacial Disorder

Non-coding mutations at the far end of a large gene desert surrounding the SOX9 gene result in a human craniofacial disorder called Pierre Robin sequence (PRS). Leveraging a human stem cell differentiation model, we identify two clusters of enhancers within the PRS-associated region that regulate SOX9 expression during a restricted window of facial progenitor development at distances up to 1.45 Mb. Enhancers within the 1.45 Mb cluster exhibit highly synergistic activity that is dependent on the Coordinator motif. Using mouse models, we demonstrate that PRS phenotypic specificity arises from the convergence of two mechanisms: confinement of Sox9 dosage perturbation to developing facial structures through context-specific enhancer activity and heightened sensitivity of the lower jaw to Sox9 expression reduction. Overall, we characterize the longest-range human enhancers involved in congenital malformations, directly demonstrate that PRS is an enhanceropathy, and illustrate how small changes in gene expression can lead to morphological variation

Matched sizes of activating and inhibitory receptor/ligand pairs are required for optimal signal integration by human Natural Killer cells

It has been suggested that receptor-ligand complexes segregate or co-localise within immune synapses according to their size, and this is important for receptor signaling. Here, we set out to test the importance of receptor-ligand complex dimensions for immune surveillance of target cells by human Natural Killer (NK) cells. NK cell activation is regulated by integrating signals from activating receptors, such as NKG2D, and inhibitory receptors, such as KIR2DL1. Elongating the NKG2D ligand MICA reduced its ability to trigger NK cell activation. Conversely, elongation of KIR2DL1 ligand HLA-C reduced its ability to inhibit NK cells. Whereas normal-sized HLA-C was most effective at inhibiting activation by normal-length MICA, only elongated HLA-C could inhibit activation by elongated MICA. Moreover, HLA-C and MICA that were matched in size co-localised, whereas HLA-C and MICA that were different in size were segregated. These results demonstrate that receptor-ligand dimensions are important in NK cell recognition, and suggest that optimal integration of activating and inhibitory receptor signals requires the receptor-ligand complexes to have similar dimensions

Talking about Gypsies: the notion of discourse as control.

The file attached to this record is the authors final peer reviewed version. The final published version can be found on the publisher's website. http://www.informaworld.com/Gypsies and Travellers are increasingly part of a debate by politicians and the media in the U.K. This discourse is not a benign reflection of events; instead it is part of a complex mechanism of control. There is a difficult relationship between the settled and travelling communities which inhibits political discussion of a strategy for site provision. In this context, the paper examines the links between discourse and control, by paying attention to Foucaultian notions of the ‘gaze’, amongst other explanations. Drawing on findings from analysis of the media, focus groups with Travellers and a case study in one local authority planning consultation exercise, the paper proposes a theoretical explanation for the link between the discourse used around Gypsies and Travellers and the control that is exercised over them, particularly in inhibiting their right to a travelling lifestyle