Beyond the script: on the inclusion of film within a discursive analysis and the immaculate construction of Daniel Blake

Modern UK welfare reforms, such as employment and support allowance (ESA), aim to provide financial support to individuals unable to work due to disability or chronic health conditions. As part of a media framing analysis, which explores the ways in which these welfare reforms have been reported by the UK press, Ken Loach’s 2016 film I, Daniel Blake was identified by the research team as an important discursive resource. Through a novel multimodal discourse analysis (which contains ‘spoilers’ for the film), we examine the ‘immaculate construction’ of fictional ESA claimant Daniel Blake and consider the implications for ‘real life’ ESA claimants. We suggest that, whilst presenting a compassionate narrative which highlights the dehumanisation experienced by ESA claimants, the film raises questions that may further problematise notions of who (and who is not) able to be considered as ‘genuinely’ deserving of welfare

Effective elimination of laser interference fringing in fluorescence microscopy by spinning azimuthal incidence angle

Laser illumination used in both conventional widefield epi-fluorescence as well as in total internal reflection fluorescence (TIRF) microscopy is subject to nonuniformities in intensity that obscure true image details. These intensity variations are interference fringes arising from coherent light scattering and diffraction at every surface in the laser light's optical path, including the lenses, mirrors, and coverslip. We present an inexpensive technique for effectively eliminating these interference fringes based upon introduction of the excitation laser beam by oblique through-the-objective incidence coupled with rapid azimuthal rotation of the plane of incidence. Although this rotation can be accomplished in several ways, a particularly simple method applicable to a free laser beam is to use an optical wedge, spun on a motor, which diverts the beam into a hollow cone of fixed angle. A system of lenses converts this collimated beam cone into a focused spot that traces a circle at the objective's back focal plane. Consequently, a collimated beam with fixed polar angle and spinning azimuthal angle illuminates the sample. If the wedge is spun rapidly, then the different interference patterns at every particular azimuthal incidence angle average out over a single camera exposure to produce an effectively uniform field of illumination. Microsc. Res. Tech., 2006. © 2006 Wiley-Liss, Inc.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/55799/1/20334_ftp.pd

Development of functional connectivity during adolescence:A longitudinal study using an action-observation paradigm

Successful interpersonal interactions rely on an ability to read the emotional states of others and to modulate one's own behavior in response. The actions of others serve as valuable social stimuli in this respect, offering the observer an insight into the actor's emotional state. Social cognition continues to mature throughout adolescence. Here we assess longitudinally the development of functional connectivity during early adolescence within two neural networks implicated in social cognition: one network of brain regions consistently engaged during action observation and another one associated with mentalizing. Using fMRI, we reveal a greater recruitment of the social-emotional network during the observation of angry hand actions in male relative to female adolescents. These findings are discussed in terms of known sex differences in adolescent social behavior

Stepwise Acquisition of Pyrimethamine Resistance in the Malaria Parasite

The spread of high-level pyrimethamine resistance in Africa threatens to curtail the therapeutic lifetime of antifolate antimalarials. We studied the possible evolutionary pathways in the evolution of pyrimethamine resistance using an approach in which all possible mutational intermediates were created by site-directed mutagenesis and assayed for their level of drug resistance. The coding sequence for dihydrofolate reductase (DHFR) from the malaria parasite Plasmodium falciparum was mutagenized, and tests were carried out in Escherichia coli under conditions in which the endogenous bacterial enzyme was selectively inhibited. We studied 4 key amino acid replacements implicated in pyrimethamine resistance: N51I, C59R, S108N, and I164L. Using empirical estimates of the mutational spectrum in P. falciparum and probabilities of fixation based on the relative levels of resistance, we found that the predicted favored pathways of drug resistance are consistent with those reported in previous kinetic studies, as well as DHFR polymorphisms observed in natural populations. We found that 3 pathways account for nearly 90% of the simulated realizations of the evolution of pyrimethamine resistance. The most frequent pathway (S108N and then C59R, N51I, and I164L) accounts for more than half of the simulated realizations. Our results also suggest an explanation for why I164L is detected in Southeast Asia and South America, but not at significant frequencies in Africa.Organismic and Evolutionary Biolog

Inhibition of the \u3cem\u3edapE\u3c/em\u3e-Encoded \u3cem\u3eN\u3c/em\u3e-Succinyl- ʟ, ʟ-diaminopimelic Acid Desuccinylase from \u3cem\u3eNeisseria meningitidis\u3c/em\u3e by ʟ-Captopril

Binding of the competitive inhibitor ʟ-captopril to the dapE-encoded N-succinyl-ʟ, ʟ-diaminopimelic acid desuccinylase from Neisseria meningitidis (NmDapE) was examined by kinetic, spectroscopic, and crystallographic methods. ʟ-Captopril, an angiotensin-converting enzyme (ACE) inhibitor, was previously shown to be a potent inhibitor of the DapE from Haemophilus influenzae (HiDapE) with an IC50 of 3.3 μM and a measured Ki of 1.8 μM and displayed a dose-responsive antibiotic activity toward Escherichia coli. ʟ-Captopril is also a competitive inhibitor of NmDapE with a Ki of 2.8 μM. To examine the nature of the interaction of ʟ-captopril with the dinuclear active site of DapE, we have obtained electron paramagnetic resonance (EPR) and magnetic circular dichroism (MCD) data for the enzymatically hyperactive Co(II)-substituted forms of both HiDapE and NmDapE. EPR and MCD data indicate that the two Co(II) ions in DapE are antiferromagnetically coupled, yielding an S = 0 ground state, and suggest a thiolate bridge between the two metal ions. Verification of a thiolate-bridged dinuclear complex was obtained by determining the three-dimensional X-ray crystal structure of NmDapE in complex with ʟ-captopril at 1.8 Å resolution. Combination of these data provides new insights into binding of ʟ-captopril to the active site of DapE enzymes as well as important inhibitor–active site residue interaction’s. Such information is critical for the design of new, potent inhibitors of DapE enzymes

Leveraging hierarchical self-assembly pathways for realizing colloidal photonic crystals

Colloidal open crystals are attractive materials, especially for their photonic applications. Self-assembly appeals as a bottom-up route for structure fabrication, but self-assembly of colloidal open crystals has proven to be elusive for their mechanical instability due to being low-coordinated. For such a bottom-up route to yield a desired colloidal open crystal, the target structure is required to be thermodynamically favored for designer building blocks and also kinetically accessible via self- assembly pathways in preference to metastable structures. Additionally, the selection of a particular polymorph poses a challenge for certain much sought-after colloidal open crystals for their applications as photonic crystals. Here, we devise hierarchical self-assembly pathways, which, starting from designer triblock patchy particles, yield in a cascade of well-separated associations first tetrahedral clusters and then tetrastack crystals. The designed pathways avoid trapping into an amorphous phase. Our analysis reveals how such a two-stage self-assembly pathway via tetrahedral clusters promotes crystallization by suppressing five- and seven-membered rings that hinder the emergence of the ordered structure. We also find that slow annealing promotes a bias toward the cubic polymorph relative to the hexagonal counterpart. Finally, we calculate the photonic band structures, showing that the cubic polymorph exhibits a complete photonic band gap for the dielectric filling fraction directly realizable from the designer triblock patchy particles. Unexpectedly, we find that the hexagonal polymorph also supports a complete photonic band gap, albeit only for an increased filling fraction, which can be realized via postassembly processing

Early Parental Positive Behavior Support and Childhood Adjustment: Addressing Enduring Questions with New Methods

A large literature provides strong empirical support for the influence of parenting on child outcomes. The current study addresses enduring research questions testing the importance of early parenting behavior to children's adjustment. Specifically, we developed and tested a novel multi‐method observational measure of parental positive behavior support at age 2. Next, we tested whether early parental positive behavior support was related to child adjustment at school age, within a multi‐agent and multi‐method measurement approach and design. Observational and parent‐reported data from mother–child dyads (N = 731; 49 percent female) were collected from a high‐risk sample at age 2. Follow‐up data were collected via teacher report and child assessment at age 7.5. The results supported combining three different observational methods to assess positive behavior support at age 2 within a latent factor. Further, parents' observed positive behavior support at age 2 predicted multiple types of teacher‐reported and child‐assessed problem behavior and competencies at 7.5 years old. Results supported the validity and predictive capability of a multi‐method observational measure of parenting and the importance of a continued focus on the early years within preventive interventions.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/110827/1/sode12103.pd

Bone turnover and metabolite responses to exercise in people with and without long-duration type 1 diabetes: a case-control study

INTRODUCTION: Exercise acutely alters markers of bone resorption and formation. As risk of fracture is increased in patients with type 1 diabetes, understanding if exercise-induced bone turnover is affected within this population is prudent. We assessed bone turnover responses to acute exercise in individuals with long-duration type 1 diabetes and matched controls. RESEARCH DESIGN AND METHODS: Participants with type 1 diabetes (n=15; age: 38.7±13.3; glycosylated hemoglobin: 60.5±6.7 mmol/mol; diabetes duration: 19.3±11.4 years) and age-matched, fitness-matched, and body mass index-matched controls (n=15) completed 45 min of incline walking (60% peak oxygen uptake). Blood samples were collected at baseline and immediately, 30 min, and 60 min postexercise. Markers of bone resorption (β-C-terminal cross-linked telopeptide of type 1 collagen, β-CTx) and formation (procollagen type-1 amino-terminal propeptide, P1NP), parathyroid hormone (PTH), phosphate, and calcium (albumin-adjusted and ionized) were measured. Data (mean±SD) were analyzed by a mixed-model analysis of variance. RESULTS: Baseline concentrations of P1NP and β-CTx were comparable between participants with type 1 diabetes and controls. P1NP did not change with exercise (p=0.20) but β-CTx decreased (p0.39). Participants with type 1 diabetes had reduced albumin and ionized calcium at all sample points (p<0.01). CONCLUSIONS: Following exercise, participants with type 1 diabetes displayed similar time-course changes in markers of bone formation and associated metabolites, but an attenuated suppression in bone resorption. The reduced albumin and ionized calcium may have implications for future bone health. Further investigation of the interactions between type 1 diabetes, differing modalities and intensities of exercise, and bone health is warranted

The p110 delta structure: mechanisms for selectivity and potency of new PI(3)K inhibitors.

Deregulation of the phosphoinositide-3-OH kinase (PI(3)K) pathway has been implicated in numerous pathologies including cancer, diabetes, thrombosis, rheumatoid arthritis and asthma. Recently, small-molecule and ATP-competitive PI(3)K inhibitors with a wide range of selectivities have entered clinical development. In order to understand the mechanisms underlying the isoform selectivity of these inhibitors, we developed a new expression strategy that enabled us to determine to our knowledge the first crystal structure of the catalytic subunit of the class IA PI(3)K p110 delta. Structures of this enzyme in complex with a broad panel of isoform- and pan-selective class I PI(3)K inhibitors reveal that selectivity toward p110 delta can be achieved by exploiting its conformational flexibility and the sequence diversity of active site residues that do not contact ATP. We have used these observations to rationalize and synthesize highly selective inhibitors for p110 delta with greatly improved potencies