Photophyiscal and Biological Implications of the Fluorophore Polymer Ratio.

Multivalent polymers can be used as gene delivery vectors, diagnostics, and therapeutics when they are conjugated with small molecules. The random nature of attaching small molecule ligands to a polymer leads to a large mixture of different ratios that can be approximated by one or more Poisson distributions. The variety of ligand to polymer ratios leads to a large difference in hydrophobic character, which can alter biological properties. The hydrophobicity ratio has been known to effect the polymer’s behavior in solution and cell. The exact effect of this ratio was determined using precise ratio materials isolated. The precise ratio of fluorophores on a polymer is specifically of interest because of the environmental dependence on fluorophore properties. The precise ratios of a fluorophore on a polymer also provides a model system for understanding how changes in hydrophobicity alter a multivalent polymer’s biological behavior. Precise dye-polymer ratios of fluorescein and TAMRA were obtained on an acetylated G5 PAMAM dendrimer by using semi-prep rp-HPLC. The fluorescein-G5 conjugates were obtained by cycloaddition of an azide functionalized fluorescein molecule with a precise ratio of cyclooctyne ligands on a G5 PAMAM dendrimer. The TAMRA-G5 PAMAM conjugates were obtained by direct conjugation and isolation. Both sets of materials were fully characterized. Amine-terminated G3 PAMAM dendrimers with precise ratios of TAMRA were also obtained and fully characterized. Similar absorption and emission spectra were obtained compared to the neutral TAMRA-G5 materials. Precise ratios of TAMRA on amine-terminated G5 PAMAM dendrimers were studied in HEK 293A cells. Dendrimers with 2-4 TAMRA were found to have greater uptake than dendrimers with a single TAMRA. FLIM showed different ratios of TAMRA alone was sufficient to change the fluorescence lifetime of the material observed inside cells. Multiple distributions of small molecules to the polymer will occur when conjugating a fluorophore and a targeting ligand or drug. Using amine terminated precise ratio TAMRA-G5 PAMAM material, controlling a second distribution of small molecules on a polymer was investigated. Multiple strategies are discussed along with the advantages and disadvantages of each strategy.PHDChemistryUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/113434/1/cadough_1.pd

Isolation and Characterization of Precise Dye/Dendrimer Ratios

Fluorescent dyes are commonly conjugated to nanomaterials for imaging applications using stochastic synthesis conditions that result in a Poisson distribution of dye/particle ratios and therefore a broad range of photophysical and biodistribution properties. We report the isolation and characterization of generation 5 poly(amidoamine) (G5 PAMAM) dendrimer samples containing 1, 2, 3, and 4 fluorescein (FC) or 6‐carboxytetramethylrhodamine succinimidyl ester (TAMRA) dyes per polymer particle. For the fluorescein case, this was achieved by stochastically functionalizing dendrimer with a cyclooctyne “click” ligand, separation into sample containing precisely defined “click” ligand/particle ratios using reverse‐phase high performance liquid chromatography (RP‐HPLC), followed by reaction with excess azide‐functionalized fluorescein dye. For the TAMRA samples, stochastically functionalized dendrimer was directly separated into precise dye/particle ratios using RP‐HPLC. These materials were characterized using 1 H and 19 F NMR spectroscopy, RP‐HPLC, UV/Vis and fluorescence spectroscopy, lifetime measurements, and MALDI. High definition : Two approaches for the formation of generation 5 PAMAM samples containing precise dye/dendrimer ratios are presented. The first approach, using direct separation based on dye hydrophobicity, generated a set of TAMRA‐containing dendrimers, and the second, using click chemistry, generated a set of fluorescein‐containing dendrimer (see figure).Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/106970/1/chem_201304854_sm_miscellaneous_information.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/106970/2/4638_ftp.pd

A Neutron Star with a Massive Progenitor in Westerlund 1

We report the discovery of an X-ray pulsar in the young, massive Galactic star cluster Westerlund 1. We detected a coherent signal from the brightest X-ray source in the cluster, CXO J164710.2-455216, during two Chandra observations on 2005 May 22 and June 18. The period of the pulsar is 10.6107(1) s. We place an upper limit to the period derivative of Pdot<2e-10 s/s, which implies that the spin-down luminosity is Edot<3e33 erg/s. The X-ray luminosity of the pulsar is L_X = 3(+10,-2)e33 (D/5 kpc)^2 erg/s, and the spectrum can be described by a kT = 0.61+/-0.02 keV blackbody with a radius of R_bb = 0.27+/-0.03 (D/5 kpc}) km. Deep infrared observations reveal no counterpart with K1 Msun. Taken together, the properties of the pulsar indicate that it is a magnetar. The rarity of slow X-ray pulsars and the position of CXO J164710.2-455216 only 1.6' from the core of Westerlund 1 indicates that it is a member of the cluster with >99.97% confidence. Westerlund 1 contains 07V stars with initial masses M_i=35 Msun and >50 post-main-sequence stars that indicate the cluster is 4+/-1 Myr old. Therefore, the progenitor to this pulsar had an initial mass M_i>40 Msun. This is the most secure result among a handful of observational limits to the masses of the progenitors to neutron stars.Comment: 4 pages, 5 figures. Final version to match ApJL (added one figure since v2

Differences in trait impulsivity indicate diversification of dog breeds into working and show lines

Impulsiveness describes the inability to inhibit behaviour in the presence of salient cues. Trait-level impulsivity exists on a continuum and individual differences can be adaptive in different contexts. While breed related differences in behavioural tendency in the domestic dog (Canis familiaris) are well established, the phenomenon within lines of a breed which have been selected more recently is not well studied, although it may challenge the popular notion of breed-typical behaviour. We describe differences in impulsivity between and within two dog breeds with working and show lines selected for different levels of impulsivity: Border Collies (herding work) and Labrador Retrievers (gun work). Recent show line selection might have lessened differences in impulsivity between breeds. We tested this hypothesis on a dataset of 1161 individuals assessed using a validated psychometric tool (Dog Impulsivity Assessment Scale - DIAS). Collies were more impulsive on average, consistent with the original purpose of breed selection. Regarding line, working Collies differed from working Labradors, but show lines from the two breeds were not significantly different. Altered or relaxed artificial selection for behavioural traits when appearance rather than behaviour become the primary focus for breeders may reduce average differences in impulsivity between breeds in show lines

Inhibition, Reinforcement Sensitivity and Temporal Information Processing in ADHD and ADHD+ODD: Evidence of a Separate Entity?

This study compared children with ADHD-only, ADHD+ODD and normal controls (age 8–12) on three key neurocognitive functions: response inhibition, reinforcement sensitivity, and temporal information processing. The goal was twofold: (a) to investigate neurocognitive impairments in children with ADHD-only and children with ADHD+ODD, and (b) to test whether ADHD+ODD is a more severe from of ADHD in terms of neurocognitive performance. In Experiment 1, inhibition abilities were measured using the Stop Task. In Experiment 2, reinforcement sensitivity and temporal information processing abilities were measured using a Timing Task with both a reward and penalty condition. Compared to controls, children with ADHD-only demonstrated impaired inhibitory control, showed more time underestimations, and showed performance deterioration in the face of reward and penalty. Children with ADHD+ODD performed in-between children with ADHD-only and controls in terms of inhibitory controls and the tendency to underestimate time, but were more impaired than controls and children with ADHD-only in terms of timing variability. In the face of reward and penalty children with ADHD+ODD improved their performance compared to a neutral condition, in contrast to children with ADHD-only. In the face of reward, the performance improvement in the ADHD+ODD group was disproportionally larger than that of controls. Taken together the findings suggest that, in terms of neurocognitive functioning, comorbid ADHD+ODD is a substantial different entity than ADHD-only

Determinants of recovery from post-COVID-19 dyspnoea: analysis of UK prospective cohorts of hospitalised COVID-19 patients and community-based controls

Background The risk factors for recovery from COVID-19 dyspnoea are poorly understood. We investigated determinants of recovery from dyspnoea in adults with COVID-19 and compared these to determinants of recovery from non-COVID-19 dyspnoea. Methods We used data from two prospective cohort studies: PHOSP-COVID (patients hospitalised between March 2020 and April 2021 with COVID-19) and COVIDENCE UK (community cohort studied over the same time period). PHOSP-COVID data were collected during hospitalisation and at 5-month and 1-year follow-up visits. COVIDENCE UK data were obtained through baseline and monthly online questionnaires. Dyspnoea was measured in both cohorts with the Medical Research Council Dyspnoea Scale. We used multivariable logistic regression to identify determinants associated with a reduction in dyspnoea between 5-month and 1-year follow-up. Findings We included 990 PHOSP-COVID and 3309 COVIDENCE UK participants. We observed higher odds of improvement between 5-month and 1-year follow-up among PHOSP-COVID participants who were younger (odds ratio 1.02 per year, 95% CI 1.01–1.03), male (1.54, 1.16–2.04), neither obese nor severely obese (1.82, 1.06–3.13 and 4.19, 2.14–8.19, respectively), had no pre-existing anxiety or depression (1.56, 1.09–2.22) or cardiovascular disease (1.33, 1.00–1.79), and shorter hospital admission (1.01 per day, 1.00–1.02). Similar associations were found in those recovering from non-COVID-19 dyspnoea, excluding age (and length of hospital admission). Interpretation Factors associated with dyspnoea recovery at 1-year post-discharge among patients hospitalised with COVID-19 were similar to those among community controls without COVID-19. Funding PHOSP-COVID is supported by a grant from the MRC-UK Research and Innovation and the Department of Health and Social Care through the National Institute for Health Research (NIHR) rapid response panel to tackle COVID-19. The views expressed in the publication are those of the author(s) and not necessarily those of the National Health Service (NHS), the NIHR or the Department of Health and Social Care. COVIDENCE UK is supported by the UK Research and Innovation, the National Institute for Health Research, and Barts Charity. The views expressed are those of the authors and not necessarily those of the funders