Preferential utilization of NADPH as the endogenous electron donor for NAD(P)H:quinone oxidoreductase 1 (NQO1) in intact pulmonary arterial endothelial cells

The goal was to determine whether endogenous cytosolic NAD(P)H:quinone oxidoreductase 1 (NQO1) preferentially uses NADPH or NADH in intact pulmonary arterial endothelial cells in culture. The approach was to manipulate the redox status of the NADH/NAD+ and NADPH/NADP+ redox pairs in the cytosolic compartment using treatment conditions targeting glycolysis and the pentose phosphate pathway alone or with lactate, and to evaluate the impact on the intact cell NQO1 activity. Cells were treated with 2-deoxyglucose, iodoacetate, or epiandrosterone in the absence or presence of lactate, NQO1 activity was measured in intact cells using duroquinone as the electron acceptor, and pyridine nucleotide redox status was measured in total cell KOH extracts by high-performance liquid chromatography. 2-Deoxyglucose decreased NADH/NAD+ and NADPH/NADP+ ratios by 59 and 50%, respectively, and intact cell NQO1 activity by 74%; lactate restored NADH/NAD+, but not NADPH/NADP+ or NQO1 activity. Iodoacetate decreased NADH/NAD+ but had no detectable effect on NADPH/NADP+ or NQO1 activity. Epiandrosterone decreased NQO1 activity by 67%, and although epiandrosterone alone did not alter the NADPH/NADP+ or NADH/NAD+ ratio, when the NQO1 electron acceptor duroquinone was also present, NADPH/NADP+ decreased by 84% with no impact on NADH/NAD+. Duroquinone alone also decreased NADPH/NADP+ but not NADH/NAD+. The results suggest that NQO1 activity is more tightly coupled to the redox status of the NADPH/NADP+ than NADH/NAD+ redox pair, and that NADPH is the endogenous NQO1 electron donor. Parallel studies of pulmonary endothelial transplasma membrane electron transport (TPMET), another redox process that draws reducing equivalents from the cytosol, confirmed previous observations of a correlation with the NADH/NAD+ ratio

Characterization of theThreshold for NAD(P)H:quinone Oxidoreductase Activity in Intact Sulforaphane-treated Pulmonary Arterial Endothelial Cells

Treatment of bovine pulmonary arterial endothelial cells in culture with the phase II enzyme inducer sulforaphane (5 μM, 24 h; sulf-treated) increased cell-lysate NAD(P)H:quinone oxidoreductase (NQO1) activity by 5.7 ± 0.6 (mean ± SEM)-fold, but intact-cell NQO1 activity by only 2.8 ± 0.1-fold compared to control cells. To evaluate the hypothesis that the threshold for sulforaphane-induced intact-cell NQO1 activity reflects a limitation in the capacity to supply NADPH at a sufficient rate to drive all the induced NQO1 to its maximum activity, total KOH-extractable pyridine nucleotides were measured in cells treated with duroquinone to stimulate maximal NQO1 activity. NQO1 activation increased NADP+ in control and sulf-treated cells, with the effect more pronounced in the sulf-treated cells, in which the NADPH was also decreased. Glucose-6-phosphate dehydrogenase (G-6-PDH) inhibition partially blocked NQO1 activity in control and sulf-treated cells, but G-6-PDH overexpression via transient transfection with the human cDNA alleviated neither the restriction on intact sulf-treated cell NQO1 activity nor the impact on the NADPH/NADP+ ratios. Intracellular ATP levels were not affected by NQO1 activation in control or sulf-treated cells. An increased dependence on extracellular glucose and a rightward shift in the Km for extracellular glucose were observed in NQO1-stimulated sulf-treated vs control cells. The data suggest that glucose transport in the sulf-treated cells may be insufficient to support the increased metabolic demand for pentose phosphate pathway-generated NADPH as an explanation for the NQO1 threshold

Experimental Demonstration of Squeezed State Quantum Averaging

We propose and experimentally demonstrate a universal quantum averaging process implementing the harmonic mean of quadrature variances. The harmonic mean protocol can be used to efficiently stabilize a set of fragile squeezed light sources with statistically fluctuating noise levels. The averaged variances are prepared probabilistically by means of linear optical interference and measurement induced conditioning. We verify that the implemented harmonic mean outperforms the standard arithmetic mean strategy. The effect of quantum averaging is experimentally tested both for uncorrelated and partially correlated noise sources with sub-Poissonian shot noise or super-Poissonian shot noise characteristics.Comment: 4 pages, 5 figure

Local Manufacturing - Strategic Operationalisation Of Lean Methods In Manufacturing-related Small And Medium-sized Enterprises (SME)

The economic, social and ecological crises of recent years have shown that a change in our understanding of production and value creation is necessary. A more even distribution of production capacities can promote social and economic stability. The ability to produce locally unavailable products or spare parts at short notice (e.g. manufacturing medical-products during the pandemic) avoids CO2 emissions due to transport and increases local production sovereignty and resilience. This means that production structures need to become more dynamic and responsive for spontaneous demand. Increasing local production at the place of need seems to be a solution that addresses the problems raised. When looking at currently available local production structures, small and medium-sized enterprises can be found at most locations. Unfortunately there is still a lack of productivity compared to industrial production in this enterprises. One form of overarching organisation and corporate culture that is largely established as standard in industry is the Lean Business Model. The introduction of such a strategy offers potentials for increasing the productivity and performance of a company regardless of its size. However, the methods and principles are not consistently applied in small and medium-sized enterprises. In the context of this work, studies were identified that deal with the feasibility of implementing Lean Methods in the manufacturing sector worldwide. The results were bundled into a new data model and subjected to a secondary analysis. The aim is to obtain a complete assessment of all lean implementation methods according to VDI2870 Holistic Production Systems. The suitability of the implementability in small and medium-sized enterprises is extended by an evaluation with regard to the target dimensions time, costs and quality. This creates a new possibility for strategic operationalisation of the Lean Methods for manufacturing companies

Temperature and Performance Inhomogeneities in PEM Electrolysis Stacks with Industrial Scale Cells

In this work temperature inhomogeneities and their influence on PEMWE performance of industrial-scale stacks are investigated. Three temperature differences are examined: (i) between the inlet and outlet, (ii) in-between the cells of a stack, (iii) between the cell’s solid materials and the fluids. A validated stack model for temperature and performance is presented which is used to quantify the above-mentioned temperature fields and their influences on current density distribution and cell voltages. For a chosen scenario, with current densities of 2.0 A cm−2, fluid inlet temperatures of 60 °C and flow-rates of 0.15 kg s−1m−2, peak temperature differences amount to 8.2 K along-the-channel. This relates to inhomogeneities of current density of up to 10% inside a cell and deviations of cell voltage of 9 mV in-between cells in the center of the stack and outer cells. For higher current densities these differences increase further. More homogeneous temperatures allow operation at elevated average temperatures without exceeding temperature limitations and reduce the spread of degradation mechanisms. Hence, homogenous profiles lead to a more hole-some utilization of electrolysis stacks. Therefore, the ability to homogenize via alternative operation such as higher flow-rate, higher pressure and altered routing of fluid-flow is analyzed

Ultra-cold atoms in an optical cavity: two-mode laser locking to the cavity avoiding radiation pressure

The combination of ultra-cold atomic clouds with the light fields of optical cavities provides a powerful model system for the development of new types of laser cooling and for studying cooperative phenomena. These experiments critically depend on the precise tuning of an incident pump laser with respect to a cavity resonance. Here, we present a simple and reliable experimental tuning scheme based on a two-mode laser spectrometer. The scheme uses a first laser for probing higher-order transversal modes of the cavity having an intensity minimum near the cavity's optical axis, where the atoms are confined by a magnetic trap. In this way the cavity resonance is observed without exposing the atoms to unwanted radiation pressure. A second laser, which is phase-locked to the first one and tuned close to a fundamental cavity mode drives the coherent atom-field dynamics.Comment: 7 pages, 7 figure

The Impact of Openness on Value Co-creation in Production Networks

AbstractThe increasing number and economic importance of production networks is one sign of the on-going paradigm shift from industrial production to value co-creation. This transformation can be described by using the notions of a value creation taxonomy, which is introduced in this paper and gives a structured overview of relevant aspects of the underlying conversion from top down to bottom up economics. In order to gain a deeper understanding of this transformation process, the specific design, characteristics and challenges of those networks will be investigated with regard to their time-dependence using a life cycle model.The present study contributes to a fundamental understanding of the importance of openness as a key success factor of value co-creation in production networks. It gives a systematic characterization of what is meant by “openness” concerning the value creation system, the value creation process and the value creation artifact. Furthermore, an adjusted life cycle model is presented, which may support both, assessment and configuration of openness within those networks by deriving adequate and phase-specific measures

Trusted CI Experiences in Cybersecurity and Service to Open Science

This article describes experiences and lessons learned from the Trusted CI project, funded by the US National Science Foundation to serve the community as the NSF Cybersecurity Center of Excellence. Trusted CI is an effort to address cybersecurity for the open science community through a single organization that provides leadership, training, consulting, and knowledge to that community. The article describes the experiences and lessons learned of Trusted CI regarding both cybersecurity for open science and managing the process of providing centralized services to a broad and diverse community.Comment: 8 pages, PEARC '19: Practice and Experience in Advanced Research Computing, July 28-August 1, 2019, Chicago, IL, US