The Pancreatic ß-Cell: A Bioenergetic Perspective.

The pancreatic ß-cell secretes insulin in response to elevated plasma glucose. This review applies an external bioenergetic critique to the central processes of glucose-stimulated insulin secretion, including glycolytic and mitochondrial metabolism, the cytosolic adenine nucleotide pool, and its interaction with plasma membrane ion channels. The control mechanisms responsible for the unique responsiveness of the cell to glucose availability are discussed from bioenergetic and metabolic control standpoints. The concept of coupling factor facilitation of secretion is critiqued, and an attempt is made to unravel the bioenergetic basis of the oscillatory mechanisms controlling secretion. The need to consider the physiological constraints operating in the intact cell is emphasized throughout. The aim is to provide a coherent pathway through an extensive, complex, and sometimes bewildering literature, particularly for those unfamiliar with the field.

Data-driven design of thin-film optical systems using deep active learning.

A deep learning aided optimization algorithm for the design of flat thin-film multilayer optical systems is developed. The authors introduce a deep generative neural network, based on a variational autoencoder, to perform the optimization of photonic devices. This algorithm allows one to find a near-optimal solution to the inverse design problem of creating an anti-reflective grating, a fundamental problem in material science. As a proof of concept, the authors demonstrate the method's capabilities for designing an anti-reflective flat thin-film stack consisting of multiple material types. We designed and constructed a dielectric stack on silicon that exhibits an average reflection of 1.52 %, which is lower than other recently published experiments in the engineering and physics literature. In addition to its superior performance, the computational cost of our algorithm based on the deep generative model is much lower than traditional nonlinear optimization algorithms. These results demonstrate that advanced concepts in deep learning can drive the capabilities of inverse design algorithms for photonics. In addition, the authors develop an accurate regression model using deep active learning to predict the total reflectivity for a given optical system. The surrogate model of the governing partial differential equations can then be broadly used in the design of optical systems and to rapidly evaluate their behavior.

Diagnostics and Treatments of COVID-19: A Living Systematic Review of Economic Evaluations.

OBJECTIVES: As healthcare systems continue to respond to the COVID-19 pandemic, cost-effectiveness evidence will be needed to identify which tests and treatments for COVID-19 offer value for money. We sought to review economic evaluations of diagnostic tests and treatments for COVID-19, critically appraising the methodological approaches used and reporting cost-effectiveness estimates, using a "living" systematic review approach. METHODS: Key databases (including MEDLINE, EconLit, Embase) were last searched on July 12, 2021. Gray literature and model repositories were also searched. Only full economic evaluations published in English were included. Studies were quality assessed and data were extracted into standard tables. Results were narratively summarized. The review was completed by 2 reviewers independently, with disagreements resolved through discussion with a senior reviewer. RESULTS: Overall, 3540 records were identified, with 13 meeting the inclusion criteria. After quality assessment, 6 were excluded because of very severe limitations. Of the 7 studies included, 5 were cost-utility analyses and 2 were cost-effectiveness analyses. All were model-based analyses. A total of 5 evaluated treatments (dexamethasone, remdesivir, hypothetical) and 2 evaluated hypothetical testing strategies. Cost-effectiveness estimates were sensitive to the treatment effect on survival and hospitalization, testing speed and accuracy, disease severity, and price. CONCLUSIONS: Presently, there are few economic evaluations for COVID-19 tests and treatments. They suggest treatments that confer a survival benefit and fast diagnostic tests may be cost effective. Nevertheless, studies are subject to major evidence gaps and take inconsistent analytical approaches. The evidence may improve for planned updates of this "living" review.

Monte Carlo-transformed field expansion method for simulating electromagnetic wave scattering by multilayered random media.

We present an efficient numerical method for simulating the scattering of electromagnetic fields by a multilayered medium with random interfaces. The elements of this algorithm, the Monte Carlo-transformed field expansion method, are (i) an interfacial problem formulation in terms of impedance-impedance operators, (ii) simulation by a high-order perturbation of surfaces approach (the transformed field expansions method), and (iii) efficient computation of the wave field for each random sample by forward and backward substitutions. Our perturbative formulation permits us to solve a sequence of linear problems featuring an operator that is deterministic, and its LU decomposition matrices can be reused, leading to significant savings in computational effort. With an extensive set of numerical examples, we demonstrate not only the robust and high-order accuracy of our scheme for small to moderate interface deformations, but also how Padé summation can be used to address large deviations.

On the consistent choice of effective permittivity and conductivity for modeling graphene.

Graphene has transformed the fields of plasmonics and photonics, and become an indispensable component for devices operating in the terahertz to mid-infrared range. Here, for instance, graphene surface plasmons can be excited, and their extreme interfacial confinement makes them vastly effective for sensing and detection. The rapid, robust, and accurate numerical simulation of optical devices featuring graphene is of paramount importance and many groups appeal to Black-Box Finite Element solvers. While accurate, these are quite computationally expensive for problems with simplifying geometrical features such as multiple homogeneous layers, which can be recast in terms of interfacial (rather than volumetric) unknowns. In either case, an important modeling consideration is whether to treat the graphene as a material of small (but non-zero) thickness with an effective permittivity, or as a vanishingly thin sheet of current with an effective conductivity. In this contribution we ponder the correct relationship between the effective conductivity and permittivity of graphene, and propose a new relation which is based upon a concrete mathematical calculation that appears to be missing in the literature. We then test our new model both in the case in which the interface deformation is non-trivial, and when there are two layers of graphene with non-flat interfacial deformation.

Launching graphene surface plasmon waves with vanishingly small periodic grating structures.

Graphene is now a crucial component of many device designs in electronics and optics. Just like the noble metals, this single layer of carbon atoms in a honeycomb lattice can support surface plasmons, which are central to several sensing technologies in the mid-infrared regime. As with classical metal plasmons, periodic corrugations in the graphene sheet itself can be used to launch these surface waves; however, as graphene plasmons are tightly confined, the role of unwanted surface roughness, even at a nanometer scale, cannot be ignored. In this work, we revisit our previous numerical experiments on metal plasmons launched by vanishingly small grating structures, with the addition of graphene to the structure. These simulations are conducted with a recently devised, rapid, and robust high-order spectral scheme of the authors, and with it we carefully demonstrate how the plasmonic response of a perfectly flat sheet of graphene can be significantly altered with even a tiny corrugation (on the order of merely 5 nm). With these results, we demonstrate the primary importance of fabrication techniques that produce interfaces whose deviations from flat are on the order of angstroms.

The singular patient in patient-centred care: physiotherapists' accounts of treatment of patients with chronic muscle pain.

A patient-centred approach has gained increasing interest in medicine and other health sciences. Whereas there are discussions about the meaning of a patient-centred approach and what the concept entails, little is known about how the patient as a person is understood in patient-centred care. This article investigates understandings of the patient as a self in patient-centred care through physiotherapy of patients with chronic muscle pain. The material consists of interviews with five Norwegian physiotherapists working in a rehabilitation clinic. Drawing on Kristeva's discussion of subjectivity in medical discourse, the study highlights two different treatment storylines that were closely entwined. One storyline focuses on open singular healing processes in which the treatment was based on openness to a search for meaning and sharing. In this storyline, the "person" at the centre of care was not essentialised in terms of biological mechanisms, but rather considered as a vulnerable, irrational and moving self. By contrast, the second storyline focused on goal-oriented interventions aimed at restoring the patient to health. Here, the person in the centre of the treatment was shaped according to model narratives about "the successful patient"; the empowered, rational, choosing and self-managing individual. As such, the findings revealed two conflicting concepts of the individual patient inherent in patient-centred care. On the one hand, the patient is seen as being a person in constant movement, and on the other, they are captured by more standardised terms designed to focus on a more stable notion of outcome of illness. Therefore, our study suggests that the therapists' will to recognise the individual in patient-centred care had a counterpart involving a marginalisation of the singular.

Nutrient and microbial water quality of the upper Ganga River, India: identification of pollution sources.

The Ganga River is facing mounting environmental pressures due to rapidly increasing human population, urbanisation, industrialisation and agricultural intensification, resulting in worsening water quality, ecological status and impacts on human health. A combined inorganic chemical, algal and bacterial survey (using flow cytometry and 16S rRNA gene sequencing) along the upper and middle Ganga (from the Himalayan foothills to Kanpur) was conducted under pre-monsoon conditions. The upper Ganga had total phosphorus (TP) and total dissolved nitrogen concentrations of less than 100 µg l-1 and 1.0 mg l-1, but water quality declined at Kannauj (TP = 420 µg l-1) due to major nutrient pollution inputs from human-impacted tributaries (principally the Ramganga and Kali Rivers). The phosphorus and nitrogen loads in these two tributaries and the Yamuna were dominated by soluble reactive phosphorus and ammonium, with high bacterial loads and large numbers of taxa indicative of pathogen and faecal organisms, strongly suggesting sewage pollution sources. The high nutrient concentrations, low flows, warm water and high solar radiation resulted in major algal blooms in the Kali and Ramganga, which greatly impacted the Ganga. Microbial communities were dominated by members of the Phylum Proteobacteria, Bacteriodetes and Cyanobacteria, with communities showing a clear upstream to downstream transition in community composition. To improve the water quality of the middle Ganga, and decrease ecological and human health risks, future mitigation must reduce urban wastewater inputs in the urbanised tributaries of the Ramganga, Kali and Yamuna Rivers.

Quantifying intracellular rates of glycolytic and oxidative ATP production and consumption using extracellular flux measurements.

Partitioning of ATP generation between glycolysis and oxidative phosphorylation is central to cellular bioenergetics but cumbersome to measure. We describe here how rates of ATP generation by each pathway can be calculated from simultaneous measurements of extracellular acidification and oxygen consumption. We update theoretical maximum ATP yields by mitochondria and cells catabolizing different substrates. Mitochondrial P/O ratios (mol of ATP generated per mol of [O] consumed) are 2.73 for oxidation of pyruvate plus malate and 1.64 for oxidation of succinate. Complete oxidation of glucose by cells yields up to 33.45 ATP/glucose with a maximum P/O of 2.79. We introduce novel indices to quantify bioenergetic phenotypes. The glycolytic index reports the proportion of ATP production from glycolysis and identifies cells as primarily glycolytic (glycolytic index > 50%) or primarily oxidative. The Warburg effect is a chronic increase in glycolytic index, quantified by the Warburg index. Additional indices quantify the acute flexibility of ATP supply. The Crabtree index and Pasteur index quantify the responses of oxidative and glycolytic ATP production to alterations in glycolysis and oxidative reactions, respectively; the supply flexibility index quantifies overall flexibility of ATP supply; and the bioenergetic capacity quantifies the maximum rate of total ATP production. We illustrate the determination of these indices using C2C12 myoblasts. Measurement of ATP use revealed no significant preference for glycolytic or oxidative ATP by specific ATP consumers. Overall, we demonstrate how extracellular fluxes quantitatively reflect intracellular ATP turnover and cellular bioenergetics. We provide a simple spreadsheet to calculate glycolytic and oxidative ATP production rates from raw extracellular acidification and respiration data.

The contributions of respiration and glycolysis to extracellular acid production.

BACKGROUND: The rate at which cells acidify the extracellular medium is frequently used to report glycolytic rate, with the implicit assumption that conversion of uncharged glucose or glycogen to lactate(-)+H(+) is the only significant source of acidification. However, another potential source of extracellular protons is the production of CO2 during substrate oxidation: CO2 is hydrated to H2CO3, which then dissociates to HCO3(-)+H(+). METHODS: O2 consumption and pH were monitored in a popular platform for measuring extracellular acidification (the Seahorse XF Analyzer). RESULTS: We found that CO2 produced during respiration caused almost stoichiometric release of H(+) into the medium. With C2C12 myoblasts given glucose, respiration-derived CO2 contributed 34% of the total extracellular acidification. When glucose was omitted or replaced by palmitate or pyruvate, this value was 67-100%. Analysis of primary cells, cancer cell lines, stem cell lines, and isolated synaptosomes revealed contributions of CO2-produced acidification that were usually substantial, ranging from 3% to 100% of the total acidification rate. CONCLUSION: Measurement of glycolytic rate using extracellular acidification requires differentiation between respiratory and glycolytic acid production. GENERAL SIGNIFICANCE: The data presented here demonstrate the importance of this correction when extracellular acidification is used for quantitative measurement of glycolytic flux to lactate. We describe a simple way to correct the measured extracellular acidification rate for respiratory acid production, using simultaneous measurement of oxygen consumption rate. SUMMARY STATEMENT: Extracellular acidification is often assumed to result solely from glycolytic lactate production, but respiratory CO2 also contributes. We demonstrate that extracellular acidification by myoblasts given glucose is 66% glycolytic and 34% respiratory and describe a method to differentiate these sources.

Launching surface plasmon waves via vanishingly small periodic gratings.

The scattering of electromagnetic waves by periodic layered media plays a crucial role in many applications in optics and photonics, in particular in nanoplasmonics for topics as diverse as extraordinary optical transmission, photonic crystals, metamaterials, and surface plasmon resonance biosensing. With these applications in mind, we focus on surface plasmon resonances excited in the context of insulator-metal structures with a periodic, corrugated interface. The object of this contribution is to study the geometric limits required to generate these fundamentally important phenomena. For this we use the robust, rapid, and highly accurate field expansions method to investigate these delicate phenomena and demonstrate how very small perturbations (e.g., a 5 nm deviation on a 530 nm period grating) can generate strong (in this instance 20%) plasmonic absorption, and vanishingly small perturbations (e.g., a 1 nm deviation on a 530 nm period grating) can generate nontrivial (in this instance 1%) plasmonic absorption.

Calcium modulation of exocytosis-linked plasma membrane potential oscillations in INS-1 832/13 cells.

In the presence of high glucose or pyruvate, INS-1 832/13 insulinoma cells undergo stochastic oscillations in plasma membrane potential (Δψp) leading to associated fluctuations in cytosolic free Ca(2+) concentration ([Ca(2+)]c). Oscillations are not driven by upstream metabolic fluctuations, but rather by autonomous ionic mechanisms, the details of which are unclear. We have investigated the nature of the oscillator, with simultaneous fluorescence monitoring of Δψp, [Ca(2+)]c and exocytosis at single-cell resolution, combined with analysis of the occurrence, frequency and amplitude of Δψp oscillations. Oscillations were closely coupled to exocytosis, indicated by coincident synaptopHluorin fluorescence enhancement. L-type Ca(2+) channel inhibitors enhanced Δψp and [Ca(2+)]c oscillation frequency in the presence of pyruvate, but abolished the sustained [Ca(2+)]c response following KCl depolarization. The L-type Ca(2+) channel inhibitor isradipine did not inhibit oscillation-linked exocytosis. The T-type Ca(2+) channel inhibitor NNC 55-0396 inhibited Δψp and [Ca(2+)]c oscillations, implying that T-type Ca(2+) channels trigger oscillations and consequent exocytosis. Since distinct ion channels operate in oscillating and non-oscillating cells, quantitative analysis of Δψp and [Ca(2+)]c oscillations in a ß-cell population may help to improve our understanding of the link between metabolism and insulin secretion.

Chronic high glucose and pyruvate levels differentially affect mitochondrial bioenergetics and fuel-stimulated insulin secretion from clonal INS-1 832/13 cells.

Glucotoxicity in pancreatic ß-cells is a well established pathogenetic process in type 2 diabetes. It has been suggested that metabolism-derived reactive oxygen species perturb the ß-cell transcriptional machinery. Less is known about altered mitochondrial function in this condition. We used INS-1 832/13 cells cultured for 48 h in 2.8 mm glucose (low-G), 16.7 mm glucose (high-G), or 2.8 mm glucose plus 13.9 mm pyruvate (high-P) to identify metabolic perturbations. High-G cells showed decreased responsiveness, relative to low-G cells, with respect to mitochondrial membrane hyperpolarization, plasma membrane depolarization, and insulin secretion, when stimulated acutely with 16.7 mm glucose or 10 mm pyruvate. In contrast, high-P cells were functionally unimpaired, eliminating chronic provision of saturating mitochondrial substrate as a cause of glucotoxicity. Although cellular insulin content was depleted in high-G cells, relative to low-G and high-P cells, cellular functions were largely recovered following a further 24-h culture in low-G medium. After 2 h at 2.8 mm glucose, high-G cells did not retain increased levels of glycolytic or TCA cycle intermediates but nevertheless displayed increased glycolysis, increased respiration, and an increased mitochondrial proton leak relative to low-G and high-P cells. This notwithstanding, titration of low-G cells with low protonophore concentrations, monitoring respiration and insulin secretion in parallel, showed that the perturbed insulin secretion of high-G cells could not be accounted for by increased proton leak. The present study supports the idea that glucose-induced disturbances of stimulus-secretion coupling by extramitochondrial metabolism upstream of pyruvate, rather than exhaustion from metabolic overload, underlie glucotoxicity in insulin-producing cells.

Pharmacological characterization of AZD5069, a slowly reversible CXC chemokine receptor 2 antagonist.

In normal physiologic responses to injury and infection, inflammatory cells enter tissue and sites of inflammation through a chemotactic process regulated by several families of proteins, including inflammatory chemokines, a family of small inducible cytokines. In neutrophils, chemokines chemokine (CXC motif) ligand 1 (CXCL1) and CXCL8 are potent chemoattractants and activate G protein-coupled receptors CXC chemokine receptor 1 (CXCR1) and CXCR2. Several small-molecule antagonists of CXCR2 have been developed to inhibit the inflammatory responses mediated by this receptor. Here, we present the data describing the pharmacology of AZD5069 [N-(2-(2,3-difluorobenzylthio)-6-((2R,3S)-3,4-dihydroxybutan-2-yloxy)[2,4,5,6-(13)C4, 1,3-(15)N2]pyrimidin-4-yl)azetidine-1-sulfonamide,[(15)N2,(13)C4]N-(2-(2,3-difluoro-6-[3H]-benzylthio)-6-((2R,3S)-3,4-dihydroxybutan-2-yloxy)pyrimidin-4-yl)azetidine-1-sulfonamide], a novel antagonist of CXCR2. AZD5069 was shown to inhibit binding of radiolabeled CXCL8 to human CXCR2 with a pIC50 value of 9.1. Furthermore, AZD5069 inhibited neutrophil chemotaxis, with a pA2 of approximately 9.6, and adhesion molecule expression, with a pA2 of 6.9, in response to CXCL1. AZD5069 was a slowly reversible antagonist of CXCR2 with effects of time and temperature evident on the pharmacology and binding kinetics. With short incubation times, AZD5069 appeared to have an antagonist profile with insurmountable antagonism of calcium response curves. This behavior was also observed in vivo in an acute lipopolysaccharide-induced lung inflammation model. Altogether, the data presented here show that AZD5069 represents a novel, potent, and selective CXCR2 antagonist with potential as a therapeutic agent in inflammatory conditions.

Mitochondrial bioenergetics and neuronal survival modelled in primary neuronal culture and isolated nerve terminals.

Mitochondria play multiple roles in the maintenance of neuronal function under physiological and pathological conditions. In addition to ATP generation, they can act as major short-term calcium sinks and can both generate, and be damaged by, reactive oxygen species. Two complementary preparations have been extensively employed to investigate in situ neuronal mitochondrial bioenergetics, primary neuronal cultures and acutely isolated nerve terminals, synaptosomes. A major focus of the cell culture preparation has been the investigation of glutamate excitotoxicity. Oxidative phosphorylation, calcium transport and reactive oxygen species play complex interlocking roles in the life and death of the glutamate exposed neuron. Synaptosomes may be isolated from specific brain regions at any developmental stage and therefore provide a valuable ex vivo approach in studying mouse models. Recent advances have allowed synaptosomal bioenergetics to be studied on a microgram scale, and, in combination with approaches to correct for functional and transmitter heterogeneity, have allowed hypotheses concerning presynaptic mitochondrial dysfunction to be tested on a variety of genetic models of neurodegenerative disorders.

Discovery and evaluation of a novel monocyclic series of CXCR2 antagonists.

Antagonism of the chemokine receptor CXCR2 has been proposed as a strategy for the treatment of inflammatory diseases such as arthritis, chronic obstructive pulmonary disease and asthma. Earlier series of bicyclic CXCR2 antagonists discovered at AstraZeneca were shown to have low solubility and poor oral bioavailability. In this Letter we describe the design, synthesis and characterisation of a new series of monocyclic CXCR2 antagonists with improved solubility and good pharmacokinetic profiles.

Method of field expansions for vector electromagnetic scattering by layered periodic crossed gratings.

In many applications of scientific and engineering interest the accurate modeling of scattering of linear waves by periodic layered media plays a crucial role. From geophysics and oceanography to materials science and imaging, the ability to simulate such configurations numerically in a rapid and robust fashion is of paramount importance. In this contribution we focus upon the specific problem of vector electromagnetic radiation interacting with a multiply layered periodic crossed diffraction grating. While all of the classical methods for the numerical simulation of partial differential equations have been brought to bear upon this problem, we argue here that in this particular context a high-order perturbation of surfaces approach is superior. In particular, we describe how the method of field expansions can be extended to the fully vectorial and three-dimensional scattering problem in the presence of multiple layers. With specific numerical experiments we will show the remarkable efficiency, fidelity, and high-order accuracy one can achieve with an implementation of this algorithm.

Re/creating entrepreneurs of the self: discourses of worker and employee 'value' and current vocational rehabilitation practices.

Vocational rehabilitation for people experiencing work disability is a social practice often situated within health services, but the social and political drivers and effects of this practice are rarely critically analysed in health research or policy. In this study we used a Foucauldian theoretical perspective to analyse the ways in which current vocational rehabilitation practices in New Zealand re/produce notions of worker and employee 'value', and how different approaches to vocational rehabilitation deploy current discourses about value. We also consider the subject positions produced through these different approaches and the identities and actions they make possible for people experiencing work disability. The analysis showed that notions about the importance of worker and employee value in a job market are pervasive in vocational rehabilitation, and reflect wider societal discourses. However, the deployment of those discourses in different approaches to vocational rehabilitation practice are diverse, producing different opportunities and constraints for people experiencing disability. We argue that an examination of these various opportunities and constraints at the level of practice approaches is important, as considerable time and resources are allocated to developing solutions to help those who do not thrive in the current systems, yet we rarely critique the premises on which the systems are based.