The hydrogen peroxide-sensitive proteome of the chloroplast in vitro and in vivo

Muthuramalingam M, Matros A, Scheibe R, Mock H-P, Dietz K-J. The hydrogen peroxide-sensitive proteome of the chloroplast in vitro and in vivo. Frontiers in Plant Science. 2013;4:54-1-54-14.Hydrogen peroxide (H2O2) evolves during cellular metabolism and accumulates under various stresses causing serious redox imbalances. Many proteomics studies aiming to identify proteins sensitive to H2O2 used concentrations that were above the physiological range. Here the chloroplast proteins were subjected to partial oxidation by exogenous addition of H2O2 equivalent to 10% of available protein thiols which allowed for the identification of the primary targets of oxidation. The chosen redox proteomic approach employed differential labeling of non-oxidized and oxidized thiols using sequential alkylation with N-ethylmaleimide and biotin maleimide. The in vitro identified proteins are involved in carbohydrate metabolism, photosynthesis, redox homeostasis, and nitrogen assimilation. By using methyl viologen that induces oxidative stress in vivo, mostly the same primary targets of oxidation were identified and several oxidation sites were annotated. Ribulose-1,5-bisphosphate (RubisCO) was a primary oxidation target. Due to its high abundance, RubisCO is suggested to act as a chloroplast redox buffer to maintain a suitable redox state, even in the presence of increased reactive oxygen species release. 2-cysteine peroxiredoxins (2-Cys Prx) undergo redox-dependent modifications and play important roles in antioxidant defense and signaling. The identification of 2-Cys Prx was expected based on its high affinity to H2O2 and is considered as a proof of concept for the approach. Targets of Trx, such as phosphoribulokinase, glyceraldehyde-3-phosphate dehydrogenase, transketolase, and sedoheptulose-1,7-bisphosphatase have at least one regulatory disulfide bridge which supports the conclusion that the identified proteins undergo reversible thiol oxidation. In conclusion, the presented approach enabled the identification of early targets of H2O2 oxidation within the cellular proteome under physiological experimental conditions

Anti-money laundering and counter-terrorist financing in the Luxembourg investment fund market

The field of Anti-Money Laundering and Counter-Terrorist Financing in their current form have had an important impact on the financial world for almost half a century. Today, Money Laundering and Terrorist Financing pose a threat to the integrity of the financial markets and systems worldwide. The intention behind implementing a regulatory Anti-Money Laundering and Counter-Terrorist Financing framework is to cut off the financial resources of criminals and to follow back the traces that financial transactions leave to the backers of the criminal organizations. Luxembourg, being the second largest center for investment funds in the world and the leading one in Europe, is dependent on its good reputation, a cornerstone of its success. Based on this, the risk exposure of the Luxembourg Fund Market with respect to Money Laundering and Terrorist Financing shall be assessed and a risk assessment for an investment fund established. Firstly, a general introduction to Money Laundering and Terrorist Financing is provided, which is complemented by a chapter specifying details of each criminal offence, differences and similarities, and the threats and harms both crimes pose. Secondly, the legal definitions and local obligations of the market participants are presented. Thirdly, the Luxembourg Fund Market in terms of figures, products and its participants in investment fund structure are addressed. In a next step, the summary of three expert interviews is presented. The questions asked are based on the information, statements and findings in the previous chapters. All information obtained is then used to establish a general risk assessment of an investment fund and to draw the final conclusion. The results show that the inherent risk of the Luxembourg Fund Market is concentrated on Money Laundering rather than on Terrorist Financing. The residual risk is completely dependent on the measures implemented by the single structures and therefore cannot really be quantified. It is very much dependent on the risk appetite of the market participants, their controls in place and the enforcement of the rules, especially when it comes to cross-border business

Polypropylene pipe interface strength on marine sandy soils with varying coarse fraction

The interface shear strength of polypropylene pipeline coatings and marine sandy soils was investigated through direct and surface-over-soil interface shear box testing. Polypropylene specimens were acquired by removal from existing manufactured steel pipes and test soils were fabricated to closely resemble typical compositions and particle size distributions of North Sea marine sediments. The test sands varied according to their coarse particle fractions, with 0, 15 and 35% being retained on a 0·4 mm sieve. Testing was carried out at the very low stresses pertinent to pipeline interfaces, between 2·5 and 37·5 kPa, in both loose and dense states. The experimental results suggest a dependency of the interface shear strength on the stress level and relative density, with the coarse particle fraction playing a modest role. Surface characterisation and lack of volumetric deformation suggests that the shearing kinematic is predominantly grain sliding rather than rolling. Interface efficiency was largely constant despite some scatter due to variability in surface specimens. The distinct seams apparent on some of the polypropylene surfaces as inherent manufacturing artefacts had a negligible influence on interface strength. The relationship between interface strength, normalised roughness and Shore D hardness was assessed, discussed and compared with results from other works

Regulation of gene expression by photosynthetic signals triggered through modified CO(2 )availability

BACKGROUND: To coordinate metabolite fluxes and energy availability, plants adjust metabolism and gene expression to environmental changes through employment of interacting signalling pathways. RESULTS: Comparing the response of Arabidopsis wild-type plants with that of the mutants adg1, pgr1 and vtc1 upon altered CO(2)-availability, the regulatory role of the cellular energy status, photosynthetic electron transport, the redox state and concentration of ascorbate and glutathione and the assimilatory force was analyzed in relation to the transcript abundance of stress-responsive nuclear encoded genes and psaA and psbA encoding the reaction centre proteins of photosystem I and II, respectively. Transcript abundance of Bap1, Stp1, psaA and psaB was coupled with seven metabolic parameters. Especially for psaA and psaB, the complex analysis demonstrated that the assumed PQ-dependent redox control is subordinate to signals linked to the relative availability of 3-PGA and DHAP, which define the assimilatory force. For the transcripts of sAPx and Csd2 high correlations with the calculated redox state of NADPH were observed in pgr1, but not in wild-type, suggesting that in wild-type plants signals depending on thylakoid acidification overlay a predominant redox-signal. Strongest correlation with the redox state of ascorbate was observed for 2CPA, whose transcript abundance regulation however was almost insensitive to the ascorbate content demonstrating dominance of redox regulation over metabolite sensing. CONCLUSION: In the mutants, signalling pathways are partially uncoupled, demonstrating dominance of metabolic control of photoreaction centre expression over sensing the redox state of the PQ-pool. The balance between the cellular redox poise and the energy signature regulates sAPx and Csd2 transcript abundance, while 2CPA expression is primarily redox-controlled

Improving Inference of Gaussian Mixtures Using Auxiliary Variables

Expanding a lower-dimensional problem to a higher-dimensional space and then projecting back is often beneficial. This article rigorously investigates this perspective in the context of finite mixture models, namely how to improve inference for mixture models by using auxiliary variables. Despite the large literature in mixture models and several empirical examples, there is no previous work that gives general theoretical justification for including auxiliary variables in mixture models, even for special cases. We provide a theoretical basis for comparing inference for mixture multivariate models with the corresponding inference for marginal univariate mixture models. Analytical results for several special cases are established. We show that the probability of correctly allocating mixture memberships and the information number for the means of the primary outcome in a bivariate model with two Gaussian mixtures are generally larger than those in each univariate model. Simulations under a range of scenarios, including misspecified models, are conducted to examine the improvement. The method is illustrated by two real applications in ecology and causal inference

Contrastive Language-Image Pretrained Models are Zero-Shot Human Scanpath Predictors

Understanding the mechanisms underlying human attention is a fundamental challenge for both vision science and artificial intelligence. While numerous computational models of free-viewing have been proposed, less is known about the mechanisms underlying task-driven image exploration. To address this gap, we present CapMIT1003, a database of captions and click-contingent image explorations collected during captioning tasks. CapMIT1003 is based on the same stimuli from the well-known MIT1003 benchmark, for which eye-tracking data under free-viewing conditions is available, which offers a promising opportunity to concurrently study human attention under both tasks. We make this dataset publicly available to facilitate future research in this field. In addition, we introduce NevaClip, a novel zero-shot method for predicting visual scanpaths that combines contrastive language-image pretrained (CLIP) models with biologically-inspired neural visual attention (NeVA) algorithms. NevaClip simulates human scanpaths by aligning the representation of the foveated visual stimulus and the representation of the associated caption, employing gradient-driven visual exploration to generate scanpaths. Our experimental results demonstrate that NevaClip outperforms existing unsupervised computational models of human visual attention in terms of scanpath plausibility, for both captioning and free-viewing tasks. Furthermore, we show that conditioning NevaClip with incorrect or misleading captions leads to random behavior, highlighting the significant impact of caption guidance in the decision-making process. These findings contribute to a better understanding of mechanisms that guide human attention and pave the way for more sophisticated computational approaches to scanpath prediction that can integrate direct top-down guidance of downstream tasks

Function and Regulation of Chloroplast Peroxiredoxin IIE

Peroxiredoxins (PRX) are thiol peroxidases that are highly conserved throughout all biological kingdoms. Increasing evidence suggests that their high reactivity toward peroxides has a function not only in antioxidant defense but in particular in redox regulation of the cell. Peroxiredoxin IIE (PRX-IIE) is one of three PRX types found in plastids and has previously been linked to pathogen defense and protection from protein nitration. However, its posttranslational regulation and its function in the chloroplast protein network remained to be explored. Using recombinant protein, it was shown that the peroxidatic Cys121 is subjected to multiple posttranslational modifications, namely disulfide formation, S-nitrosation, S-glutathionylation, and hyperoxidation. Slightly oxidized glutathione fostered S-glutathionylation and inhibited activity in vitro. Immobilized recombinant PRX-IIE allowed trapping and subsequent identification of interaction partners by mass spectrometry. Interaction with the 14-3-3 υ protein was confirmed in vitro and was shown to be stimulated under oxidizing conditions. Interactions did not depend on phosphorylation as revealed by testing phospho-mimicry variants of PRX-IIE. Based on these data it is proposed that 14-3-3υ guides PRX‑IIE to certain target proteins, possibly for redox regulation. These findings together with the other identified potential interaction partners of type II PRXs localized to plastids, mitochondria, and cytosol provide a new perspective on the redox regulatory network of the cell

Reactive oxygen species and redox regulation in mesophyll and bundle sheath cells of C4 plants

Turkan I, Uzilday B, Dietz K-J, Bräutigam A, Ozgur R. Reactive oxygen species and redox regulation in mesophyll and bundle sheath cells of C4 plants. Journal of Experimental Botany. 2018;69(14):3321-3331.Redox regulation, antioxidant defence, and reactive oxygen species (ROS) signalling are critical in performing and tuning metabolic activities. However, our concepts have mostly been developed for C3 plants since Arabidopsis thaliana has been the major model for research. Efforts to convert C3 plants to C4 to increase yield (such as IRRI’s C4 Rice Project) entail a better understanding of these processes in C4 plants. Various photosynthetic enzymes that take part in light reactions and carbon reactions are regulated via redox components, such as thioredoxins as redox transmitters and peroxiredoxins. Hence, understanding redox regulation in the mesophyll and bundle sheath chloroplasts of C4 plants is of paramount importance: it appears impossible to utilize efficient C4 photosynthesis without understanding its exact redox needs and the regulation mechanisms used during light reactions. In this review, we discuss current knowledge on redox regulation in C3 and C4 plants, with special emphasis on the mesophyll and bundle sheath differences that are found in C4. In these two cell types in C4 plants, linear and cyclic electron transport in the chloroplasts operate differentially when compared to C3 chloroplasts, changing the redox needs of the cell. Therefore, our focus is on photosynthetic light reactions, ROS production dynamics, antioxidant defence, and thiol-based redox regulation, with the aim of providing an overview of our current knowledge