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The hydrogenase maturase HydG produces multiple equivalents of free CO and CN - during catalysis. This production correlates with the activation of hydrogenase, supporting a model in which free CO and CN - are relevant to maturation.
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Abstract In plastids, conversion of light energy into ATP relies on cytochrome f, a key electron carrier with a heme covalently attached to a CXXCH motif. Covalent heme attachment requires reduction of the disulfide-bonded CXXCH by CCS5 and CCS4. CCS5 receives electrons from the oxidoreductase CCDA, while CCS4 is a protein of unknown function. In Chlamydomonas reinhardtii, loss of CCS4 or CCS5 yields a partial cytochrome f assembly defect. Here, we report that the ccs4ccs5 double mutant displays a synthetic photosynthetic defect characterized by a complete loss of holocytochrome f assembly. This defect is chemically corrected by reducing agents, confirming the placement of CCS4 and CCS5 in a reducing pathway. CCS4-like proteins occur in the green lineage, and we show that HCF153, a distant ortholog from Arabidopsis thaliana, can substitute for Chlamydomonas CCS4. Dominant suppressor mutations mapping to the CCS4 gene were identified in photosynthetic revertants of the ccs4ccs5 mutants. The suppressor mutations yield changes in the stroma-facing domain of CCS4 that restore holocytochrome f assembly above the residual levels detected in ccs5. Because the CCDA protein accumulation is decreased specifically in the ccs4 mutant, we hypothesize the suppressor mutations enhance the supply of reducing power through CCDA in the absence of CCS5. We discuss the operation of a CCS5-dependent and a CCS5-independent pathway controlling the redox status of the heme-binding cysteines of apocytochrome f.
Blockchain technology is a relatively novel technology that can be used to develop more decentralized, autonomous and tamper-evident solutions. A feature that can aid Transactive Energy Systems to reach their goals by enabling individual actors to communicate and reach consensus with other participants in a more decentralized fashion. However, technical barriers to evaluate and adopt this type of technology within the electrical domain still exist. To facilitate this task, BLOSEM Unified Testing Platform (UTP), a DOE-sponsored, multi-lab effort intends to accelerate the development of solutions by offering a common set of reusable services that can be used to interconnect existent grid tools with blockchain services. UTP is intended to serve as development platform that can provide application engineers with the technical means to evaluate potential blockchain solutions, by enabling them to concentrate on the actual application functionalities while at the same time abstracting the connectivity and performance measurement tasks. The use of BLOSEM UTP is further demonstrated by implementing two potential use cases that are intended to validate both the feasibility of implementing these applications as blockchain-based solutions while also demonstrating the features provided by UTP.
Young exoplanets can offer insight into the evolution of planetary atmospheres, compositions, and architectures. We present the discovery of the young planetary system TOI 451 (TIC 257605131, Gaia DR2 4844691297067063424). TOI 451 is a member of the 120 Myr old Pisces–Eridanus stream (Psc–Eri). We confirm membership in the stream with its kinematics, its lithium abundance, and the rotation and UV excesses of both TOI 451 and its wide-binary companion, TOI 451 B (itself likely an M-dwarf binary). We identified three candidate planets transiting in the Transiting Exoplanet Survey Satellite data and followed up the signals with photometry from Spitzer and ground-based telescopes. The system comprises three validated planets at periods of 1.9, 9.2, and 16 days, with radii of 1.9, 3.1, and 4.1 R {sub ⨂}, respectively. The host star is near-solar mass with V = 11.0 and H = 9.3 and displays an infrared excess indicative of a debris disk. The planets offer excellent prospects for transmission spectroscopy with the Hubble Space Telescope and the James Webb Space Telescope, providing the opportunity to study planetary atmospheres that may still be in the process of evolving.
The pore structure characteristics of shales are controlled by their mineralogical and organic matter (OM) composition. However, the contributions by different components in shales at varying thermal maturities remain poorly understood. In this study, Devonian New Albany Shale and Marcellus Shale samples spanning a thermal maturity from marginally mature (vitrinite reflectance R o 0.55%) to post-mature (R o 2.41 %) were selected to study the control of composition on the pore structure properties of shales at different stages of thermal maturation. Scanning electron microscope (SEM) imaging was used to examine pore types in shales, and low-pressure N 2 and CO 2 adsorption analyses were used to quantitatively characterize the mesopore and micropore characteristics of bulk shales and major components in shales. The results show that matrix-associated pores including interparticle pores between silt-sized mineral grains, phyllosilicate framework pores, and intraparticle pores within mineral grains exist in all samples but become less common with increasing maturity, which is likely caused by elevated compaction, cementation, and occlusion with bitumen. Secondary organic pores were not observed under SEM at marginal maturity but were detected in the condensate–wet gas and dry gas windows, with more organic pores in the dry gas window. At marginal maturity, OM has large amounts of mesopores and micropores as demonstrated by low-pressure N 2 and CO 2 adsorption analyses of OM isolated from shales, even though no OM-hosted pores were observed under SEM. With increasing thermal maturity, the mesopore and micropore specific surface area of OM increase and make greater contributions to the pore structure properties of bulk shales. The mesopore and micropore properties of shales are controlled by OM content and maturity as well as by clay mineral type and content, and they can be estimated from the contribution of each component at different stages of thermal maturation. In conclusion, accurate evaluation of the pore volume and SSA of shales will have important implications for assessing gas adsorption and transport in shales.
Temporal partitioning of life history traits among syntopic reptiles can facilitate co-occurrence, but may be influenced by environmental factors and evolutionary history. We used 24 years of continuous capture data in the Florida sandhills to evaluate the timing and duration of hatching, maturation and/or surface activity for ten reptile species, spanning multiple clutch strategies, taxonomic relationships, and habits. We hypothesised: i) species would differ in seasonal timing of hatching and maturation; ii) hatching and maturation periods would be more seasonally-synchronised in fossorial than terrestrial or semi-aquatic reptiles; iii) monthly and annual temperature anomalies would be positively related to hatching, maturation, and surface activity anomalies, and iv) groupings of reptiles by clutch strategy, taxonomic relationship, and habit, would explain more variation in the timing and duration of hatching and maturation than species alone. Seasonal timing of response variables varied widely among species. Hatching peaked for > 1 species during most calendar months. Maturation and surface activity periods ranged from aseasonal to highly-seasonal among species. Hatching began 1.5 months earlier and was more prolonged for terrestrial than fossorial species overall. Hatching peaked in early to mid-summer for terrestrial and fossorial species, and winter for the semi-aquatic Kinosternon subrubrum. Terrestrial and fossorial species did not differ in average timing, duration, or overlap of maturation periods; semi-aquatic Liodytes pygaea matured more consistently across all seasons than other species. Monthly temperature anomalies were negatively correlated with monthly maturation for Plestiodon egregius. Annual temperature and precipitation anomalies were related to annual hatching, maturation, and surface activity trends for several species. Taxonomic relationship, habit, and species explained some variation in hatching and maturation timing and duration. Our results illustrate the influence of environment and evolutionary relationships on the timing of important life history traits. Keywords: Age class, Drift fence, Community, Interspecific, Life history
Maturity models can be used to provide government agencies, industry associations, and organizations operating nuclear facilities with the ability to quickly evaluate the maturity of their cybersecurity programs and identify areas to prioritize for improvement. The Cybersecurity Capability Maturity Model (C2M2) was developed by the U.S. Department of Energy to allow organizations in the energy sector to evaluate the programmatic capabilities of their cybersecurity programs in a consistent manner, communicate programmatic maturity information, prioritize cybersecurity investments in targeted areas of concern, and track how the maturity of their cybersecurity program evolves over time. The C2M2 is designed for use by any critical infrastructure organization regardless of ownership, structure, or size. The C2M2 can be easily fine-tuned to access the maturity of nuclear cybersecurity programs and their application at individual facilities. Built on a foundation of existing cybersecurity standards, frameworks, programs, and initiatives, the model features 10 security domains. Performance in each security domain is characterized using a structured set of cybersecurity practices that represent activities an organization can perform to improve cybersecurity in their domain. Each practice can be quickly evaluated as being either fully, largely, partially, or not implemented. Once practices are evaluated for each security domain, the model defines four maturity indicator levels that apply independently to each domain in the model. To earn a maturity level in a given domain, an organization must adequately perform all the practices for that maturity level and its predecessor level(s). A small assessment team can conduct a C2M2 assessment in a single day. A screening version of the C2M2 allows an initial look at the maturity of nuclear cybersecurity programs that could be completed in under an hour.
The purpose of this paper is to review existing smart manufacturing (SM) maturity models' dimensions and maturity levels to assess their applicability and drawbacks. There are many maturity models available but many of them have not been validated or do not provide a useful guide or tool for applications. This gap creates the need for a review of the existing maturity model's applicability. Nineteen peer-reviewed maturity models related to “Digital Transformation,” “Industry 4.0” or “Smart Manufacturing” were selected based on a systematic literature review and five consulting firm models were selected based on the author's industry knowledge. The chosen models were analyzed to determine 10 categories of dimensions. Then they are assessed on a 1–5 scale for how applicable they are in the 10 categories of dimensions. The five “consulting firm” models have a first-mover advantage, are more widely used in industry and are more applicable, but some require payment, and they lack published details and validation. The 19 “peer reviewed” models are not as widely used, lack awareness in the industry and are not as easy to apply because of no web tool for self-assessment, but they are improving. The categories defined to characterize the models and facilitate comparisons for users include “Information Technology (IT) and Cyber-Physical System (CPS) and Data,” “Strategy and Organization,” “Supply Chain and Logistics,” “Products and Services,” “Culture and Employees,” “Technology and Capabilities,” “Customer and Market,” “Cybersecurity and Risk,” “Leadership and Management” and “Governance and Compliance.” The analyzed maturity models were particularly weak in the areas of cybersecurity, leadership and governance. Researchers and practitioners can use this review with consideration of their specific needs to determine if a maturity model is applicable or if a new model needs to be developed. The review can also aid in the development of maturity models through the discussion of each of the dimension categories. Finally, compared to existing reviews of SM maturity models, this research determines comprehensive dimension categories and focuses on applicability and drawbacks.
Coastal communities are facing increased risks due to climate change, as evidenced by heightened shocks such as hurricanes and wildfires, and stressors including sea-level rise and drought. To confront these risks, many coastal communities are planning and designing their built environments in new ways, with the overall goal of increasing their resilience. However, resilience practices have been widely guided by an event-based timeframe, which may not be sufficient to confront the multiple and overlapping threats expected over the coming decades. To address this concern, this study investigated a long-term perspective on resilience progression, termed resilience maturity. To better understand resilience maturity in practice, this study drew directly on community perspectives, thereby identifying common challenges. Furthermore, variation in resilience maturity across communities provided an opportunity to identify existing successful strategies. Inductive qualitative analysis was used to examine perspectives from 15 interviews with local practitioners across 12 coastal communities. The analysis revealed that communities in early stages of resilience maturity often struggle with local stakeholder alignment and minimal dedicated resilience resources. In contrast, those further along the maturity path often face technology barriers and challenges in aligning regional stakeholders. Few examples of a fully mature resilience culture were found within the communities studied, underscoring systemic barriers to achieving advanced resilience maturity. Overall, these findings supported development of a practical framework for classifying community resilience efforts, identifying common obstacles, and informing localized resilience advancement.
The organometallic H-cluster of the [FeFe]-hydrogenase is assembled in vivo through a complex process requiring the action of three dedicated maturation enzymes, HydG, HydE, and HydF, as well as the aminomethyl-lipoyl-H-protein (H met ) of the glycine cleavage system (GCS). Here we probe the role of HydF and its [4Fe-4S] cluster in [FeFe]-hydrogenase maturation by using a defined semisynthetic approach in which [Fe I 2 (μ-SH) 2 (CO) 4 (CN) 2 ] 2– ([2Fe] E ) is used to bypass HydE and HydG, and GCS components are used in place of cell lysate. We show that inclusion of the iron–sulfur carrier protein NfuA and the high-CO-affinity myoglobin variant Mb H64L provides dramatically improved hydrogenase activities up to 828 μmol/min/mg, equivalent to the best reported activities for Chlamydomonas reinhardtii [FeFe]-hydrogenase isolated from the native organism. Apo-HydF lacking a [4Fe-4S] cluster provides very little hydrogenase activity; however, full maturation is restored with the addition of NfuA, which we demonstrate reconstitutes the [4Fe-4S] cluster of HydF. In addition, a HydF variant lacking a [4Fe-4S] cluster by changing two cysteine ligands to alanine is completely unable to support either semisynthetic maturation using [2Fe] E , or full maturation using HydG and HydE, even in the presence of NfuA, demonstrating that the HydF [4Fe-4S] cluster is absolutely essential for [FeFe]-hydrogenase maturation. The possibility that the HydF [4Fe-4S] cluster plays a role in direct binding of [2Fe] E is negated by our results with the HydF D311C variant, which demonstrate that the labile Asp311 cluster ligand is not essential for [2Fe] E binding and HydA maturation. We therefore conclude that [2Fe]E binds HydF adjacent to, but not directly coordinated to, the [4Fe-4S] cluster. The HydF [4Fe-4S] cluster is proposed to be essential due to its impact on the [2Fe] E binding orientation and the ability of the HydF/[2Fe] E complex to form productive interactions with H met or the H met /T-protein complex during DTMA ligand biosynthesis.
Earned value management systems (EVMS) are used by industry practitioners to successfully manage projects and programs; however, there has been no past study to characterize and evaluate the impact of EVMS maturity on project performance. This paper introduces a novel framework, developed to measure the EVMS maturity, composed of 56 attributes arrayed across ten subprocesses, which together make up an integrated project/program management system. This paper describes how the framework was conceived and developed, based on literature review and data collected from industry workshops. Then, it examines the relationship between EVMS maturity and ten project performance metrics through statistical analyses. The study analyzed project data from 35 projects and programs worth over $21.8 billion in total cost. For this sample, results showed statistically significant differences between projects with high and low EVMS maturity; on average, projects exhibiting high EVMS maturity have 54% less cost overruns and 19% less change orders versus their performance measurement baseline. Higher maturity projects and programs also achieve better customer satisfaction, meet business objectives more often, and are more often in compliance with EVMS standards and guidelines. This paper contributes to the body of knowledge by providing a practical and novel framework for assessing the maturity of earned value management systems. Practitioners can use the framework to improve their integrated project and program management.
The [FeFe]-hydrogenase employs an active-site 6Fe H-cluster to catalyze the reversible reduction of protons to H 2 . A [4Fe-4S] subcluster of the H-cluster is synthesized by housekeeping iron-sulfur cluster assembly machinery, and then dedicated hydrogenase maturation enzymes, together with components of the glycine cleavage system, build and deliver a [2Fe] subcluster to generate the full H-cluster. Here, we report that theEscherichia coliiron-sulfur carrier protein NfuA supports in vitro maturation of fully active [FeFe]-hydrogenase, with H 2 production rates comparable to that of the in vivo-maturedChlamydomonas reinhardtii[FeFe]-hydrogenase (CrHydA). Inclusion of NfuA in the in vitro maturation process improves its efficacy by delivering the iron essential for formation of the [Fe II (cys)(CN)(CO) 2 ] – synthon at the dangler iron site of the HydG auxiliary cluster. NfuA serves an additional role in reconstituting and maintaining the catalytically essential iron-sulfur clusters on the maturase enzymes HydE, HydF, and HydG. Further inclusion of a high CO affinity myoglobin variant (Mb H64L ) sequesters free CO generated during the maturation process, minimizing formation of the CO-inhibited H ox -CO enzyme state, significantly increasing hydrogenase activity. The addition of NfuA and Mb H64L to the fully defined maturation system thus results in an in vitro [FeFe]-hydrogenase maturation system that generates highly active enzyme while providing insights into factors important to in vivo maturation.
Maturation of [FeFe]-hydrogenase (HydA) involves synthesis of a CO, CN - , and dithiomethylamine (DTMA)-coordinated 2Fe subcluster that is inserted into HydA to make the active hydrogenase. This process requires three maturation enzymes: the radical S-adenosyl-l-methionine (SAM) enzymes HydE and HydG, and the GTPase HydF. In vitro maturation with purified maturation enzymes has been possible only when clarified cell lysate was added, with the lysate presumably providing essential components for DTMA synthesis and delivery. We report maturation of [FeFe]-hydrogenase using a fully defined system that includes components of the glycine cleavage system (GCS), but no cell lysate. Our results reveal for the first time an essential role for the aminomethyl-lipoyl-H-protein of the GCS in hydrogenase maturation and the synthesis of the DTMA ligand of the H-cluster. In addition, we show that ammonia is the source of the bridgehead nitrogen of DTMA.
Insulin secretory granule (ISG) maturation is a crucial aspect of insulin secretion and glucose homeostasis. The regulation of this maturation remains poorly understood, especially how secretory stimuli affect ISG maturity and subcellular localization. In this study, we used soft X-ray tomography (SXT) to quantitatively map ISG morphology, density, and location in single INS-1E and mouse pancreatic β cells under the effect of various secretory stimuli. We found that the activation of glucokinase (GK), gastric inhibitory polypeptide receptor (GIPR), glucagon-like peptide-1 receptor (GLP-1R), and G protein-coupled receptor 40 (GPR40) promotes ISG maturation. Each stimulus induces unique structural remodeling in ISGs, by altering size and density, depending on the specific signaling cascades activated. These distinct ISG subpopulations mobilize and redistribute in the cell, altering the overall cellular structural organization. Our results provide insight into how current diabetes and obesity therapies impact ISG maturation and may inform the development of future treatments that target maturation specifically.
Abstract Maturation of [FeFe]‐hydrogenase (HydA) involves synthesis of a CO, CN − , and dithiomethylamine (DTMA)‐coordinated 2Fe subcluster that is inserted into HydA to make the active hydrogenase. This process requires three maturation enzymes: the radical S‐adenosyl‐ l ‐methionine (SAM) enzymes HydE and HydG, and the GTPase HydF. In vitro maturation with purified maturation enzymes has been possible only when clarified cell lysate was added, with the lysate presumably providing essential components for DTMA synthesis and delivery. Here we report maturation of [FeFe]‐hydrogenase using a fully defined system that includes components of the glycine cleavage system (GCS), but no cell lysate. Our results reveal for the first time an essential role for the aminomethyl‐lipoyl‐H‐protein of the GCS in hydrogenase maturation and the synthesis of the DTMA ligand of the H‐cluster. In addition, we show that ammonia is the source of the bridgehead nitrogen of DTMA.
Shewanella oneidensis, a metal reducer and facultative anaerobe, expresses a large number of c-type cytochromes, many of which function as anaerobic reductases. All of these proteins contain the typical heme-binding motif CXXCH and require the Ccm proteins for maturation. Two c-type cytochrome reductases also possess atypical heme-binding sites, the NrfA nitrite reductase (CXXCK) and the SirA sulfte reductase (CX12NKGCH). S. oneidensis MR-1 encodes two cytochrome c synthetases (CcmF and SirE) and two apocytochrome c chaperones (CcmI and SirG). SirE located in the sir gene cluster is required for the maturation of SirA, but not NrfA. Here we show that maturation of SirA requires the combined function of the two apocytochrome c chaperones CcmI and SirG. Loss of either protein resulted in decreased sulfte reductase. Furthermore, SirA was not detected in a mutant that lacked both chaperones, perhaps due to misfolding or instability. These results suggest that CcmI interacts with SirEFG during SirA maturation, and with CcmF during maturation of NrfA. Additionally, we show that CRP regulates expression of sirA via the newly identifed transcriptional regulatory protein, SirR.
The concept of the Ad-Mat approach is to leverage existing adjacent markets across a broad scope of technologies in order to reduce the manufacturing learning curve and ultimately accelerate redox flow battery (RFB) deployment at scale. Lithium-ion batteries (LIBs) are currently the dominant energy storage technology, and they came to technological maturity under unique market conditions when there was no meaningful competition in the consumer electronic and electric vehicle (EV) space. Today, alternative chemistries that may be technologically better-suited for long-duration storage applications are experiencing a high barrier to entry. This is in large part due to the substantial bias towards the scaled-up production and supply chain that now exists for LIBs. In the case of RFBs in particular, numerous analyses have suggested that RFBs should theoretically have a much lower system cost than LIBs - however, this relies on a mature and competitive manufacturing landscape, which has been extremely challenging to achieve for both flow batteries and other LIB competitors. At the moment, LIB alternatives tend to have isolated small-scale manufacturing pathways, which preclude the economies of scale that would be required to compete with the mature LIB industry. In the present state of the industry, niche manufacturing tools and approaches have evolved to support each alternative technology, such that there is substantial replication and duplication in effort. Continuing to pursue a strategy of isolated manufacturing processes/approaches for each LIB-alternative may never allow for at-scale deployment. In order for RFBs to meaningfully compete with LIBs in the realm of LDES, a new disruptive approach based on cross-industry learning and coordination is needed - and this is exactly what our Ad-Mat concept aims to tackle. In this re-envisioned manufacturing landscape, tools and processes from mature industries can be adapted and deployed across the range of alternative energy storage technologies. Adapting tools, equipment, processes, and industrial learning from mature industries to meet the technological requirements of RFBs would open new markets for existing OEMs in adjacent industries, would prevent unnecessary duplication and re-development, would improve efficiency across the manufacturing chain, and would ultimately support reduced costs and accelerated deployment of RFBs at scale.