A guide to integrating immunohistochemistry and chemical imaging

© 2018 The Royal Society of Chemistry. Chemical imaging provides new insight into the fundamental atomic, molecular, and biochemical composition of tissue and how they are interrelated in normal physiology. Visualising and quantifying products of pathogenic reactions long before structural changes become apparent also adds a new dimension to understanding disease pathogenesis. While chemical imaging in isolation is somewhat limited by the nature of information it can provide (e.g. peptides, metals, lipids, or functional groups), integrating immunohistochemistry allows simultaneous, targeted imaging of biomolecules while also mapping tissue composition. Together, this approach can provide invaluable information on the inner workings of the cell and the molecular basis of diseases

Exercise-related sensations contribute to decrease power during repeated cycle sprints with limited influence on neural drive

Purposes: We manipulated the inspired oxygen fraction (FiO2) to examine the effects of physiological perturbations on exercise-related sensations and the neural drive of the quadriceps during repeated, brief, maximal cycle sprints. Methods: Nine active males completed a repeated sprint cycle protocol (10 × 4-s maximal sprints with 30 s of passive recovery) in normoxia (NM; FiO2 0.21) and severe normobaric hypoxia (HY; FiO2 0.13). Peak power, quadriceps Root Mean Squared electromyography (RMS EMG), physiological (heart rate, arterial oxygen saturation, blood lactate concentration) and perceptual responses were recorded. Results: The 10 sprints in HY were associated with lower arterial oxygen saturation values compared to NM (80.7 ± 0.9 vs. 95.6 ± 0.6%; P0.47). Mean power for sprints 1-10 were lower (-13 ± 3%; P=0.001; ES=0.79), and sprint decrement was more pronounced in HY compared to NM (21.4 ± 3.7 vs. 13.2 ± 2.7%; P=0.003). There was a 17% decrease in RMS EMG activity from the first to the last sprint (P<0.001; ES=0.65), independent of condition (P=0.597; ES=0.04). Conclusions: Despite severe hypoxia exacerbating both physiological and perceptual perturbations, the performance decrement observed during the repeated sprint protocol did not coincide with an accentuated decline in RMS EMG activity. These data suggest that higher-than-normal exercise-related sensations or perceptions coincide with fatigue during repeated sprinting, independent of changes in neural drive, when the task characteristics are known beforehand

Rugby-specific small-sided games training is an effective alternative to stationary cycling at reducing clinical risk factors associated with the development of type 2 diabetes: A randomized, controlled trial

© 2015 Mendham et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Introduction: The present study investigated whether rugby small-sided games (SSG) could be an effective alternative to continuous stationary cycling (CYC) training at reducing clinical risk factors associated with the development of type 2 diabetes mellitus (T2DM). Methods: Thirty-three middle-aged (48.6±6.6y), inactive men were randomized into a CYC (n=11), SSG (n=11), or control (CON, n=11) group. Participants trained 3d·wk-1 for 8 weeks, while control participants maintained normal activity and dietary patterns. Exercise duration was matched between groups, which involved CYC or SSG (four quarters, interspersed with 2- min passive recovery). Both training programs were designed to induce similar internal loads of maximal heart rate (∼80-85%HRmax) and rating of perceived exertion. Pre- and post-intervention testing included dual-energy x-ray absorptiometry scan, graded exercise test, fasting 2h oral glucose tolerance test and resting muscle biopsy. Western blotting was used to assess the content of skeletal muscle proteins associated with mitochondrial biogenesis and glucose regulation. Results: Both CYC and SSG increased VO2 at 80%HRmax, and reduced glycated haemoglobin, glucose area under the curve (AUC; SSG, -2.3±2.4; CYC -2.2±1.6 mmol·L1(120min)1; p1(120min)1; p0.05). There were no differences within or between conditions for protein content of peroxisome proliferator-activated receptor gamma coactivator-1α, sirtuin-1, p53, glucose transporter-4, protein kinase AKT/PKB, myocyte enhancer factor 2A, mitochondrial transcription factor, nuclear respiratory factor (NRF)-1, NRF-2 or mitochondrial complexes I-V (p>0.05). Conclusion: Rugby small-sided games is an effective alternative to continuous cycling for improving metabolic risk-factors associated with the prevention of T2DM. Despite such positive adaptations in clinical risk factors, there were no changes in the content of skeletal muscle proteins associated with glucose regulation and mitochondrial biogenesis. Trial Registration: Australian New Zealand Clinical Trial Registry ACTRN1261300087471

Age modulates the injury-induced metallomic profile in the brain

© 2017 The Royal Society of Chemistry. The biological transition metals iron (Fe), copper (Cu) and zinc (Zn) are thought to contribute to the neuronal pathologies that occur following traumatic brain injury (TBI), and indeed our previously published work in young (3 month-old) mice clearly demonstrates a significant spatiotemporal modulation of metals following TBI. Of note, however, is the literature observation that there is both an apparent detrimental effect of aging on TBI outcomes and an alteration in metals and their various transporters with normal advancing age. Therefore, to determine whether there was an interaction between aging, metals and TBI, we have utilised laser ablation-inductively coupled plasma-mass spectrometry to examine the spatial and temporal distribution of Fe, Zn and Cu following an acute controlled cortical impact brain injury in aged (24 months) rodents. The relative abundance of metals in corresponding regions within the ipsilateral and contralateral hemispheres as well as the hippocampus was assessed. Substantial region and time point specific alterations in Fe, Zn and Cu were identified immediately and up to 28 days post-TBI. The data from this follow-up study has also been compared to our previous data from young animals, and aged mice exhibit an appreciably enhanced and persistent elevation of all metals in every region surveyed, with individual metal disparities at various time points observed post-injury. This may potentially contribute to the acceleration in the onset of cognitive decline and neurological disease that has been observed in the aged population following head trauma

A population-based analysis of germline BAP1 mutations in melanoma

Germline mutation of the BRCA1 associated protein-1 (BAP1) gene has been linked to uveal melanoma, mesothelioma, meningioma, renal cell carcinoma and basal cell carcinoma. Germline variants have also been found in familial cutaneous melanoma pedigrees, but their contribution to sporadic melanoma has not been fully assessed. We sequenced BAP1 in 1,977 melanoma cases and 754 controls and used deubiquitinase assays, a pedigree analysis, and a histopathological review to assess the consequences of the mutations found. Sequencing revealed 30 BAP1 variants in total, of which 27 were rare (ExAc allele frequency <0.002). Of the 27 rare variants, 22 were present in cases (18 missense, one splice acceptor, one frameshift and two near splice regions) and 5 in controls (all missense). A missense change (S98R) in a case that completely abolished BAP1 deubiquitinase activity was identified. Analysis of cancers in the pedigree of the proband carrying the S98R variant and in two other pedigrees carrying clear loss-of-function alleles showed the presence of BAP1-associated cancers such as renal cell carcinoma, mesothelioma and meningioma, but not uveal melanoma. Two of these three probands carrying BAP1 loss-of-function variants also had melanomas with histopathological features suggestive of a germline BAP1 mutation. The remaining cases with germline mutations, which were predominantly missense mutations, were associated with less typical pedigrees and tumours lacking a characteristic BAP1-associated histopathological appearances, but may still represent less penetrant variants. Germline BAP1 alleles defined as loss-of-function or predicted to be deleterious/damaging are rare in melanoma

Computation in Physical Systems: A Normative Mapping Account

The relationship between abstract formal procedures and the activities of actual physical systems has proved to be surprisingly subtle and controversial, and there are a number of competing accounts of when a physical system can be properly said to implement a mathematical formalism and hence perform a computation. I defend an account wherein computational descriptions of physical systems are high-level normative interpretations motivated by our pragmatic concerns. Furthermore, the criteria of utility and success vary according to our diverse purposes and pragmatic goals. Hence there is no independent or uniform fact to the matter, and I advance the ‘anti-realist’ conclusion that computational descriptions of physical systems are not founded upon deep ontological distinctions, but rather upon interest-relative human conventions. Hence physical computation is a ‘conventional’ rather than a ‘natural’ kind

Trapped lipopolysaccharide and LptD intermediates reveal lipopolysaccharide translocation steps across the Escherichia coli outer membrane

Lipopolysaccharide (LPS) is a main component of the outer membrane of Gram-negative bacteria, which is essential for the vitality of most Gram-negative bacteria and plays a critical role for drug resistance. LptD/E complex forms a N-terminal LPS transport slide, a hydrophobic intramembrane hole and the hydrophilic channel of the barrel, for LPS transport, lipid A insertion and core oligosaccharide and O-antigen polysaccharide translocation, respectively. However, there is no direct evidence to confirm that LptD/E transports LPS from the periplasm to the external leaflet of the outer membrane. By replacing LptD residues with an unnatural amino acid p-benzoyl-L-phenyalanine (pBPA) and UV-photo-cross-linking in E.coli, the translocon and LPS intermediates were obtained at the N-terminal domain, the intramembrane hole, the lumenal gate, the lumen of LptD channel, and the extracellular loop 1 and 4, providing the first direct evidence and “snapshots” to reveal LPS translocation steps across the outer membrane

Single vortex-antivortex pair in an exciton polariton condensate

In a homogeneous two-dimensional system at non-zero temperature, although there can be no ordering of infinite range, a superfluid phase is predicted for a Bose liquid. The stabilization of phase in this superfluid regime is achieved by the formation of bound vortex-antivortex pairs. It is believed that several different systems share this common behaviour, when the parameter describing their ordered state has two degrees of freedom, and the theory has been tested for some of them. However, there has been no direct experimental observation of the phase stabilization mechanism by a bound pair. Here we present an experimental technique that can identify a single vortex-antivortex pair in a two-dimensional exciton polariton condensate. The pair is generated by the inhomogeneous pumping spot profile, and is revealed in the time-integrated phase maps acquired using Michelson interferometry, which show that the condensate phase is only locally disturbed. Numerical modelling based on open dissipative Gross-Pitaevskii equation suggests that the pair evolution is quite different in this non-equilibrium system compared to atomic condensates. Our results demonstrate that the exciton polariton condensate is a unique system for studying two-dimensional superfluidity in a previously inaccessible regime