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The development of curative treatments for mitochondrial diseases, which are often caused by mutations in mitochondrial DNA (mtDNA) that impair energy metabolism and other aspects of cellular homoeostasis, is hindered by an incomplete understanding of the underlying biology and a scarcity of cellular and animal models. Here we report the design and application of a library of double-stranded-DNA deaminase-derived cytosine base editors optimized for the precise ablation of every mtDNA protein-coding gene in the mouse mitochondrial genome. We used the library, which we named MitoKO, to produce near-homoplasmic knockout cells in vitro and to generate a mouse knockout with high heteroplasmy levels and no off-target edits. MitoKO should facilitate systematic and comprehensive investigations of mtDNA-related pathways and their impact on organismal homoeostasis, and aid the generation of clinically meaningful in vivo models of mtDNA dysfunction.

Internal variability is the dominant cause of projection uncertainty of Arctic sea ice in the short and medium term. However, it is difficult to determine the realism of simulated internal variability in climate models, as observations only provide one possible realization while climate models can provide numerous different realizations. To enable a robust assessment of simulated internal variability of Arctic sea ice, we use a resampling technique to build synthetic ensembles for both observations and climate models, focusing on interannual variability, which is the dominant time scale of Arctic sea ice internal variability. We assess the realism of the interannual variability of Arctic sea ice cover as simulated by six models from phase 5 of the Coupled Model Intercomparison Project (CMIP5) that provide large ensembles compared to four observational datasets. We augment the standard definition of model and observational consistency by representing the full distribution of resamplings, analogous to the distribution of variability that could have randomly occurred. We find that modeled interannual variability typically lies within observational uncertainty. The three models with the smallest mean state biases are the only ones consistent in the pan-Arctic for all months, but no model is consistent for all regions and seasons. Hence, choosing the right model for a given task as well as using internal variability as an additional metric to assess sea ice simulations is important. The fact that CMIP5 large ensembles broadly simulate interannual variability consistent within observational uncertainty gives confidence in the internal projection uncertainty for Arctic sea ice based on these models. Significance Statement The purpose of this study is to evaluate the historical simulated internal variability of Arctic sea ice in climate models. Determining model realism is important to have confidence in the projected sea ice evolution from these models, but so far only mean state and trends are commonly assessed metrics. Here we assess internal variability with a focus on the interannual variability, which is the dominant time scale for internal variability. We find that, in general, models agree well with observations, but as no model is within observational uncertainty for all months and locations, choosing the right model for a given task is crucial. Further refinement of internal variability realism assessments will require reduced observational uncertainty.

PurposeThe purpose of this study is to examine whether differences in occupational burnout between Black and White officers can be explained by differences in areas of worklife and differences in indicators of reform.Design/methodology/approachUsing a modified version of the Maslach Burnout Inventory, information on emotional exhaustion and depersonalization was collected from 910 Black and 8,240 White officers through the National Police Research Platform. Structural equation models with bootstrapped confidence intervals were used to test mediation hypotheses and estimate indirect effects.FindingsBlack officers reported significantly less emotional exhaustion and depersonalization than White officers. Approximately 46% of the difference in emotional exhaustion between Black and White officers was accounted for by workload, workplace control, job satisfaction, alignment with agency values, inclusivity in the department, and the rejection of a hostile mindset toward the community. Approximately 28% of the difference in depersonalization was accounted for by workload, workplace control, job satisfaction, alignment with agency values, and the rejection of a hostile mindset.Originality/valueThis study reinforces research that demonstrates that Black officers report less emotional exhaustion and depersonalization than White officers and shows that some but not all differences in burnout can be accounted for by racial differences in areas of worklife and the rejection of a hostile mindset toward the community. More research should be focused on occupational stress in law enforcement and how Black officers are adapting to and thriving in the policing profession.

Mitochondrial activity differs markedly between organs, but it is not known how and when this arises. Here we show that cell lineage-specific expression profiles involving essential mitochondrial genes emerge at an early stage in mouse development, including tissue-specific isoforms present before organ formation. However, the nuclear transcriptional signatures were not independent of organelle function. Genetically disrupting intra-mitochondrial protein synthesis with two different mtDNA mutations induced cell lineage-specific compensatory responses, including molecular pathways not previously implicated in organellar maintenance. We saw downregulation of genes whose expression is known to exacerbate the effects of exogenous mitochondrial toxins, indicating a transcriptional adaptation to mitochondrial dysfunction during embryonic development. The compensatory pathways were both tissue and mutation specific and under the control of transcription factors which promote organelle resilience. These are likely to contribute to the tissue specificity which characterizes human mitochondrial diseases and are potential targets for organ-directed treatments.

Improving the efficiency of internal combustion engines can result in lower operating costs and enable reduced total cost of vehicle ownership. On the other hand, it is of utmost importance that the development of engine technologies is also directed toward curbing climate-altering and harmful emissions. One such technology is gasoline compression ignition, which can offer increased fuel efficiency and reduced emissions and criteria pollutants. By means of modal analysis methods, this study focuses on characterizing the internal flow behavior of a production, eight-hole, medium-duty diesel injector operating with gasoline-like fuels under typical diesel injection conditions. High-resolution tomographic reconstructions of the injector tip geometry, along with needle motion measurements, were obtained via X-ray measurements. This information was combined with a previously validated computational setup to perform a series of in-nozzle flow computational fluid dynamics (CFD) simulations. The CFD results were analyzed to identify the connection between the injector’s global behavior and relevant flowfields such as velocity and vorticity. The results obtained with the X-ray geometry were then compared against those obtained with a nominal geometry to investigate the effect of the injector’s geometric features on its behavior. Variations to the K-factor and inlet ellipticity of the orifices were explored, and these parameters’ ability to reduce in-orifice cavitation was assessed by analyzing the predicted in-orifice fuel vapor volume fraction. Relatively large and persistent flow structures arising within the injector sac were identified as the main driver for orifice-to-orifice variability, as their location and size impacted the amount of fuel delivered by each orifice. Assessing the extent to which these energetic structures impact the injector performance is of interest; thus, it is desirable to investigate the most energetically significant flowfield structures as a function of modifications to specific features of the injector geometry. Modal decomposition tools, such as space-only proper orthogonal decomposition (POD) and spectral proper orthogonal decomposition (SPOD) were applied to gain insight into dominant coherent structures that persist over the early phase of the needle opening event. It was found that the geometrical modifications applied resulted in variations of the modal energy content, as well as in the leading dominant spatial mode features. Additionally, it was found that the dominant frequency associated with the most significant energetic content was largely invariant to the geometrical modifications, with the difference between realistic and modified geometries being mostly found in the modal energy content.

Accurate fuel injection modeling remains of critical importance to the simulation of gas turbine engines as the predicted spray structure dictates fuel-air mixing, combustion, and emissions in the combustor. The prefilming airblast atomizer relies on the formation of a thin conical sheet of fuel, which breaks up due to its interaction with the counter-swirling airstreams. Recent x-ray imaging performed at Argonne National Laboratory’s Advanced Photon Source of a prefilming airblast atomizer revealed the formation of a frothy film, which is comprised of both liquid and bubbles. This finding challenges the inherent assumption for existing spray models, which assume that the film is only comprised of liquid fuel, and motivates the detailed investigation of the film formation process. To investigate the physics governing the impingement of a liquid jet in crossflow on a plate, a computational study was carried out. Large Eddy Simulations (LES) coupled with an algebraic volume-of-fluid (VOF) approach were performed to model a liquid water jet interacting with a subsonic crossflow and subsequently impinging on an aluminum plate. For the condition studied, a grid convergence study revealed that a minimum cell size of 25 μm was sufficient to adequately resolve the jet-wall interaction and film formation process, with good agreement with the x-ray measurements. These simulations also predicted the formation of bubbles within the film due to the entrainment of air. The influence of adhesion on the film characteristics and bubble distribution was explored using two different contact angles representing water on polished and unpolished aluminum plates. Although an increase in contact angle was not observed to affect the average film thickness, the lower interfacial tension reduced the number of bubbles that were formed, but increased the average size.

Aircraft atomizers generally rely on multiple physical phenomena to introduce, distribute, and mix liquid fuel with the continuous air flow. Many such devices use plain jets for high liquid flow rates and film shear atomization to encourage droplet formation. To further characterize the near-field of processes, experiments were conducted to examine multiphase development of a liquid jet issuing into a subsonic crossflow. The jet impinged on a wall and the liquid subsequently filmed and convected downstream until reaching the end of the splitter plate, where it broke up by shear forces. X-ray diagnostics were used to interrogate different regions of the flow at the Advance Photon Source at Argonne National Laboratory 9-ID beamline. The projected liquid mass distribution was measured through x-ray absorption correlations using the collimated beam. The light was conditioned for Ultra-Small-Angle-X-ray-Scattering measurements, which were collected simultaneously with the transmission signal. These transmission and scattering signals were used to compute the projected mass and path-specific surface area of the spray, which were combined to calculate Satuer mean diameter. Though the transmission is typically collected using focused beam with a finer spatial resolution during a separate experimental campaign, the near-simultaneous acquisition allowed for more accurate registration between the signals and control of the operating condition. The transmission mapping confirmed the liquid path length of the unbroken liquid jet issuing into the domain and the interaction point with the wall. The spreading and flowing of the liquid down the plate and shedding from the trailing edge of the plate revealed an order of magnitude greater liquid path length, indicating the spreading of the film. X-ray scattering results indicate surface wave formation on the liquid jet and the initial stripping of droplets from the column. Furthermore, the scattering was enhanced for droplets rebounding due to splashing from the plate. The combined signals were used to calculate the droplet diameters in the shear breakup region trailing the splitter plate. These combined measurements provide detailed breakup information to inform inputs required to initialize Lagrangian spray calculations, as well as validate high-fidelity atomization simulations.

We present recent results toward the quantification of spray characteristics at engine conditions for an eight-hole counter-bored (stepped) GDI injector – Spray G in the ECN denomination. This computational study is characterized by two novel features: the detailed description of a real injector's internal surfaces via tomographic reconstruction; and a general equation of state that represents the thermodynamic properties of homogeneous liquid-vapor mixtures. The combined level-set moment-of-fluid approach, coupled to an embedded boundary formulation for moving solid walls, makes it possible to seamlessly connect the injector's internal flow to the spray. The Large Eddy Simulation (LES) discussed here presents evidence of partial hydraulic flipping and, during the closing transient, string cavitation. Results are validated by measurements of spray density profiles and droplet size distribution.

This study aimed (1) to provide estimates of total mean retention times of milk replacer (MR), concentrates, and roughage in veal calves fed a mixed diet; (2) to determine the effect of level and type of solid feed (SF) on passage kinetics of MR, concentrates, and roughages in veal calves; and (3) to compare passage kinetics in veal calves using the fecal excretion curves of indigestible markers and a noninvasive 13C tracer breath test approach to determine whether the latter technique can serve as an alternative. At the start of the trial, 48 Holstein-Friesian calves (6 wk of age; 68 ± 7.7 kg of body weight; BW) were assigned to 1 of 4 dietary treatments (for statistical analysis, only 39 calf observations were used). Three treatments contained chopped wheat straw as roughage in the SF mixture in a concentrate:roughage ratio of 90:10 (dry matter basis). The SF level was 20 g/kg of metabolic BW per day (low straw), 30 g/kg of metabolic BW per day (middle straw), or 40 g/kg of metabolic BW per day (high straw). The fourth treatment (high hay) contained long perennial ryegrass hay as roughage in the SF mixture in a concentrate:roughage ratio of 70:30 (dry matter basis, at 40 g/kg of metabolic BW per day). The quantity of MR was fixed for the high straw treatment, whereas the amount of MR for the other treatments during the adaptation period was adjusted based on a pair gain strategy (i.e., exchanging ration components but keeping similar net energy). At the end of the adaptation period, calves ranged from 12 to 15 wk of age with an average BW of 123 ± 8.6 kg. Passage kinetics of concentrates were estimated by measuring 13C enrichment excess of CO2 in breath from a pulsed-dose of [1-13C]octanoate. Passage kinetics of roughage, concentrates, and MR were also estimated using fecal excretion curves obtained after ingestion of chromium-mordanted roughage, Yb2O3, and Co-EDTA, respectively. We conclude that [1-13C]octanoate cannot serve as a measure for oro-duodenal transit of concentrates because of unrealistic estimates. Based on the fecal excretion curves, we concluded that the total mean retention time of MR (i.e., time to peak; the moment that the excretion curve reaches peak concentration) was, on average, 12.4 h, and that the passage kinetics of MR was not affected by the level or type of SF. The mean retention time of concentrates was shorter (21.4 h) than that of both straw (59.1 h) and hay (36.8 h), and was not affected by the level or type of SF. Also, the mean retention time of the slowest compartment (i.e., the rumen) was shorter for concentrates (39.6 h) than that of straw (110.0 h) and hay (59.2 h). Contrary, the passage of roughage was affected by level and type of SF. Long hay increased time to peak by 22.3 h and decreased ruminal mean retention time by 50.8 h relative to chopped straw, indicating that the passage rate of long hay is faster than that of chopped straw. We conclude that the level and type of SF only affects the passage kinetics of roughage and not that of MR and concentrates.

Maintaining the viscosity and density of drilling fluids within their optimal performance margins is of utmost importance in running a safe and efficient operation when drilling oil and gas wells. Safety and efficiency may be adversely affected by current measurement practices of these two key properties as they still mostly rely on onsite personnel to carry out periodic manual measurements. These laborious measurement protocols limit the quantity and quality of the produced data. Here we describe an automated system for viscosity/density measurements of drilling fluids based on a tuning fork electromechanical resonator. The resonator works as the frequency-defining element of an oscillator circuit, and a digital gain control algorithm is used to control the feedback loop and actuate the device. The self-oscillating circuit allows the measurement to be repeated several times per second, enabling higher measurement precision and real-time monitoring in changing flows. The resonator was integrated into an in-tank system, which performed continuous measurements at the suction tank of the mud circulating system during drilling operations. Measurements in three different wells are presented. The results were in very good agreement with manual measurements reported from standard mud reports and further highlighted opportunities in detecting operational problems from the real-time data streams obtained.

Twelve patients who had undergone costal osteochondral graft reconstruction of the proximal pole of scaphoid were evaluated with clinical examination, patient-reported outcome scores and radiographs with an average follow-up of 10 years (range 3.5–18). The range of wrist motion was not significantly changed compared with the preoperative range of motion and functional outcomes scores were acceptable. The patients reported low pain scores despite the universal presence of radiographic changes of reduced carpal height and arthritis of the midcarpal and radiocarpal joints. Costal osteochondral graft reconstruction of the proximal pole of scaphoid offers good long-term pain relief and function. Level of evidence: IV

The internal details of fuel injectors have a profound impact on the emissions from gasoline direct injection engines. However, the impact of injector design features is not currently understood, due to the difficulty in observing and modeling internal injector flows. Gasoline direct injection flows involve moving geometry, flash boiling, and high levels of turbulent two-phase mixing. In order to better simulate these injectors, five different modeling approaches have been employed to study the engine combustion network Spray G injector. These simulation results have been compared to experimental measurements obtained, among other techniques, with X-ray diagnostics, allowing the predictions to be evaluated and critiqued. The ability of the models to predict mass flow rate through the injector is confirmed, but other features of the predictions vary in their accuracy. The prediction of plume width and fuel mass distribution varies widely, with volume-of-fluid tending to overly concentrate the fuel. All the simulations, however, seem to struggle with predicting fuel dispersion and by inference, jet velocity. This shortcoming of the predictions suggests a need to improve Eulerian modeling of dense fuel jets.

It is well known that cavitation erosion in fuel injectors can prevent reliable engine performance after only several thousand hours of operation. However, current simulation tools lack the ability to link flow predictions within the fuel injector to both the efficacy of combustion strategies and lifetime of the injector. Multiphase flow simulation predictions were studied and compared between an informed baseline injector geometry and an x-ray scanned eroded injector geometry. Overall, erosion was found to decrease the fuel mass delivery and injection velocities. A two-stage static coupling approach was employed to link the predicted injection conditions from non-eroded and eroded injectors with the external spray simulations under reacting conditions. Combustion modeling in this coupled approach was carried out using the Unsteady Flamelet Progress Variable approach with a detailed chemical mechanism for n-dodecane, comprising of 2,755 species and 11,173 reactions. Erosion in the injectors led to lower rates of spray penetration in comparison to the baseline configurations. Analysis of the reacting spray simulations revealed an insensitivity of ignition to erosion, yet shorter lift off lengths, higher levels of the soot, and lower levels of NOx were predicted in the eroded injector.

SummaryThe Tricarboxylic Acid Cycle (TCA) cycle is arguably the most critical metabolic cycle in physiology and exists as an essential interface coordinating cellular metabolism, bioenergetics, and redox homeostasis. Despite decades of research, a comprehensive investigation into the consequences of TCA cycle dysfunction remains elusive. Here, we targeted two TCA cycle enzymes, fumarate hydratase (FH) and succinate dehydrogenase (SDH), and combined metabolomics, transcriptomics, and proteomics analyses to fully appraise the consequences of TCA cycle inhibition (TCAi) in kidney epithelial cells. Our comparative approach shows that TCAi elicits a convergent rewiring of redox and amino acid metabolism dependent on the activation of ATF4 and the integrated stress response (ISR). Furthermore, we also uncover a divergent metabolic response, whereby acute FHi, but not SDHi, can maintain asparagine levels via reductive carboxylation and maintenance of cytosolic aspartate synthesis. Our work highlights an important interplay between the TCA cycle, redox biology and amino acid homeostasis.HighlightsTCA cycle inhibition promotes GSH synthesis and impairs de novo aspartate and proline synthesisDisruption of mitochondrial thiol redox homeostasis phenocopies TCA cycle inhibition by promoting GSH synthesis and impairing proline and aspartate synthesisAcute FHi, but not SDHi, can maintain asparagine levels via reductive carboxylation and maintenance of cytosolic aspartate synthesisTCA cycle inhibition mimics an amino acid deprivation-type response and activates ATF4 via the integrated stress response to maintain redox and amino acid homeostasis

Empirical growth models have widespread application in the field of aquaculture. These models allow estimates of harvest size and waste outputs in addition to nutrient and feed requirements. In an effort to increase the ability to predict shrimp growth, the specific growth rate (SGR) and thermal‐unit growth coefficient (TGC) models were fitted to 15 datasets encompassing growth of Pacific whiteleg shrimp (Litopenaeus vannamei). Shrimp were reared under commercial conditions in Southeast Asia with weights ranging from 0.01 g to 34 g. Growth rates were regressed against body weights to identify changes in growth pattern across life stages. Analysis identified two distinct patterns of growth, with a break point between stanzas at 7.5 g. The body weight exponent of the TGC model, traditionally assumed to be (1 − b) = 1/3, was solved for iteratively in each identified growth stanza in an effort to improve the goodness of fit of the TGC model. Average body weight exponents in the first and second stanzas were 0.416 and 0.952 respectively. Projected growth trajectories using these exponents resulted in significantly better fits in comparison to the traditional TGC and the SGR on the basis of statistical measures of goodness of fit.

Post-transcriptional RNA modifications, the epitranscriptome, play important roles in modulating the functions of RNA species. Modifications of rRNA are key for ribosome production and function. Identification and characterization of enzymes involved in epitranscriptome shaping is instrumental for the elucidation of the functional roles of specific RNA modifications. Ten modified sites have been thus far identified in the mammalian mitochondrial rRNA. Enzymes responsible for two of these modifications have not been characterized. Here, we identify METTL15, show that it is the main N4-methylcytidine (m4C) methyltransferase in human cells and demonstrate that it is responsible for the methylation of position C839 in mitochondrial 12S rRNA. We show that the lack of METTL15 results in a reduction of the mitochondrial de novo protein synthesis and decreased steady-state levels of protein components of the oxidative phosphorylation system. Without functional METTL15, the assembly of the mitochondrial ribosome is decreased, with the late assembly components being unable to be incorporated efficiently into the small subunit. We speculate that m4C839 is involved in the stabilization of 12S rRNA folding, therefore facilitating the assembly of the mitochondrial small ribosomal subunits. Taken together our data show that METTL15 is a novel protein necessary for efficient translation in human mitochondria.

Increasing evidence suggests that variability in hepatic glucuronidation is a critical determinant of mammalian species-specific sensitivity to the Fusarium mycotoxin deoxynivalenol (DON). Concurrently, ongoing work regarding the effects of DON on fish has led to broad classifications concerning species-specific sensitivity (e.g. rainbow trout are highly sensitive to DON and channel catfish are highly tolerant to DON). This study was designed to determine if sensitivity to DON in fish is related to inherent differences in UDP-glucuronosyltransferase (UDPGT) activity and to evaluate if increased dietary digestible starch content could be an effective strategy to increase glucuronidation capacity. Rainbow trout or Nile tilapia (initial average body weight = 21.9 and 8.1 g/fish, respectively) were fed two series (12 or 24% digestible starch) of three diets containing graded levels of naturally occurring DON (0.1, 0.7 and 1.3 ppm) for 10 weeks. Significant linear decreases in weight gain, thermal-unit growth coefficient (TGC, P ≤ .05), feed efficiency (P .05) with the exception of a quadratic increase in NRE (P .05, DON × digestible starch). A significant quadratic decrease (P ≤ .05) in UDPGT activity was associated with increasing levels of DON in rainbow trout fed the diet containing 12% digestible starch, whereas trout and tilapia fed the diet containing 24% digestible starch experienced significant quadratic or linear increases (P ≤ .05) in UDPGT activity, respectively. However, UDPGT activity was approximately 10-fold higher in rainbow trout than in Nile tilapia. This suggests that UDPGT activity is not responsible for the observed species-specific sensitivity to DON or it is not an appropriate biomarker of DON glucuronidation. A significantly higher relative fold expression of CYP1A in tilapia compared to trout (P = .0001) may offer potential opportunity for future study. We demonstrate for the first time that Nile tilapia are unaffected by practically relevant levels of DON which are detrimental to rainbow trout. This study also extends the current knowledge of DON metabolism and represents an important contribution to the development of efficacious nutritional mycotoxin mitigation strategies.

Since particulate emissions control technologies are dependent on filtration technologies, development of porous materials with optimized pore structures is crucial to improve filtration efficiency and pressure drop across filters. Despite increasing attention to 3D measurements of porous materials, there are few reports of pore structure investigations of diesel/gasoline particulate filters (D/GPFs) using 3D visualization techniques. In this work, GPF 3D pore structures were examined using X-ray tomography (XRT) to identify the impacts of catalyst coating or ash loading. Voxel resolution of 2.93 µm made it impossible to distinguish coating or ash materials from the cordierite substrate or to recognize smaller pores than the voxel resolution. However, pores up to 200 µm, which are responsible for the most pore volume, were successfully analyzed. Coating and ash loading resulted in lowering average pore diameter, total porosity, and open porosity, as the peak density of pore diameter at 60 µm decreased, while pores below 20 µm increased. Also, the visualized closed pores, which were homogeneously distributed throughout the bare filter, tended to get larger from inlet to outlet sides and more likely to be on the surface inlet due to coating and ash loading, indicating gas pathways originally existing in open pores were blocked due to coating and ash loading, leading to increased pressure drop. The ash impact was found to be more noticeable on mid and back positions than on front position. In addition, the investigation of areal porosity along the direction of gas flow suggested that while ash penetration could reach the outlet side as noted from increased closed pore population, most ash particles would be contained up to 150 µm. The 2D crosscut microscopic analysis that requires destructive procedures with the limited examination area could provide underestimation of ash penetration, whereas the 3D XRT analysis seems to provide more accurate information of pore structure changes due to ash loading at different locations.

One of the major concerns in combustion engines is the sensitivity of engine performance to fuel properties. Recent works have shown that even slight differences in fuel properties can cause significant changes in performance and emission of an engine. In order to design the combustion engines with multi-fuel flexibilities, the precise assessment of fuel sensitivity on liquid jet atomization process is a prerequisite since the resulting fuel/air mixture is critical to the subsequent combustion process. The present study is focusing on the effect of physical fuel properties, mostly viscosity difference, on the breakup process of the liquid jet injected into still air. Two different jet fuels, CAT-A2 and CAT-C3, are considered here as surrogates for a fossil-based fuel and a bio-derived high-viscosity alternative fuel. The simulations are performed using the volume-of-fluid (VoF) interface tracking method coupled to Lagrangian particle method in order to capture the breakup instabilities of jets and the resulting droplets. The investigations take the actual geometry of the injector into account to resolve the unsteady flow phenomena inside the nozzle that impact the turbulence transition and atomization. The simulation results are compared to the experimental measurement using X-ray radiography. Both simulation and X-ray measurements consistently describe the effects of different fuels on the fundamental properties of atomization including the breakup length, transverse liquid volume fraction and the droplet sauter-mean-diameter. The application of a Detailed Numerical Simulation approach complemented by unique X-ray diagnostics is novel and providing new understanding and research directions in engine spray dynamics.

A new diagnostic for the quantification of Sauter mean diameter in high-pressure fuel sprays has been recently developed using combined optical and X-ray measurements at the Georgia Institute of Technology and Argonne National Laboratory, respectively. This diagnostic utilizes liquid scattering extinction measurements from diffuse back-illumination imaging, conducted at Georgia Tech, and liquid absorption measurements from X-ray radiography, conducted at Argonne’s Advanced Photon Source. The new diagnostic, entitled the scattering–absorption measurement ratio, quantifies two-dimensional distributions of path-integrated Sauter mean diameter, enabling the construction of the spatial history of drop size development within practical fuel sprays. This technique offers unique benefits over conventional drop-sizing methods in that it can be more robust in optically dense regions of the spray, while also providing high spatial resolution of the corresponding droplet field. The methodology for quantification of Sauter mean diameter distributions using the scattering–absorption measurement ratio technique has been previously introduced and demonstrated in diesel sprays using the Engine Combustion Network Spray D injector; however, a more detailed treatment of measurement uncertainties has been needed. In this work, we present a summary of the various sources of measurement uncertainty in the scattering–absorption measurement ratio diagnostic, like those due to the experimental setup, data processing methods, and theoretical assumptions, and assess how these sources of uncertainty affect the quantified Sauter mean diameter. The spatially resolved Sauter mean diameter measurements that result from the scattering–absorption measurement ratio diagnostic will be especially valuable to the engine modeling community for the quantitative validation of spray submodels in engine computational fluid dynamics codes. Careful evaluation and quantification of measurement uncertainties are important to support accurate model validation and to ensure the development of more predictive spray models.

We describe six persons from three families with three homozygous protein truncating variants in PUS7: c.89_90del (p.Thr30Lysfs(∗)20), c.1348C>T (p.Arg450(∗)), and a deletion of the penultimate exon 15. All these individuals have intellectual disability with speech delay, short stature, microcephaly, and aggressive behavior. PUS7 encodes the RNA-independent pseudouridylate synthase 7. Pseudouridylation is the most abundant post-transcriptional modification in RNA, which is primarily thought to stabilize secondary structures of RNA. We show that the disease-related variants lead to abolishment of PUS7 activity on both tRNA and mRNA substrates. Moreover, pus7 knockout in Drosophila melanogaster results in a number of behavioral defects, including increased activity, disorientation, and aggressiveness supporting that neurological defects are caused by PUS7 variants. Our findings demonstrate that RNA pseudouridylation by PUS7 is essential for proper neuronal development and function.

Mitochondria contain a specific translation machinery for the synthesis of mitochondria-encoded respiratory chain components. Mitochondrial tRNAs (mt-tRNAs) are also generated from the mitochondrial DNA and, similar to their cytoplasmic counterparts, are post-transcriptionally modified. Here, we find that the RNA methyltransferase METTL8 is a mitochondrial protein that facilitates 3-methyl-cytidine (m

Arctic summer sea ice has decreased dramatically over the last few decades, particularly in the summer months. The observed decline is faster than most CMIP5 models, but if internal variability is considered, models and observations are not inconsistent. With only one realization of reality in observations, it is difficult to disentangle the role of internal variability from the forced response. We directly compare one metric of internal variability by resampling both observations and models. So far we have compared five CMIP5 models from the CLIVAR multi-model large ensemble archive (CanESM2, CESM1, CSIRO MK36, GFDL ESM2M, and MPI ESM1). For the pan-Arctic, these models were found to have higher internal variability than observed by approximately 10-50% across models and seasons. Spatially, we find the variability in ice edge region is consistently modelled well in March. In September, although the member mean of the models shows both smaller absolute declines and smaller variation of such declines with resampling, the models have at least one member consistent with observations. This allows us to conclude that the models’ representation of this specific metric of internal variability is consistent with observations.

14 páginas, 2 tablas, 6 figuras., Gas production profiles typically show a monotonically increasing monophasic pattern. However, atypical gas production profiles exist whereby at least two consecutive phases of gas production or additional extraneous features that distort the typical profile are present. Such profiles are more likely to occur with the use of a fecal inoculum and are much less well described. The presence of multiple phases or non-descript extraneous features makes it difficult to apply directly recommended modeling approaches such as standard response functions or classical growth functions. To overcome such difficulties, extensions of the Mitscherlich equation and a numerical modeling option also based on the Mitscherlich are explored. The numerical modeling option uses an estimate of relative rate obtained from the smoothed data profile and an estimate of maximum gas produced together with any lag time information drawn from the raw data to construct a simple Mitscherlich equation. In summary, this article illustrates the analysis of atypical gas production profiles obtained using a fecal inoculum and explores the methodology of numerical modeling to reconstruct equivalent typical growth-like trends., This research was funded in part by The Canada Research Chairs program, grant number 045867 (Natural Sciences and Engineering Research Council of Canada, Ottawa).

Mitochondria contain a specific translation machinery for the synthesis of mitochondria-encoded respiratory chain components. Mitochondrial tRNAs (mt-tRNAs) are also generated from the mitochondrial DNA and, similar to their cytoplasmic counterparts, are post-transcriptionally modified. Here, we find that the RNA methyltransferase METTL8 is a mitochondrial protein that facilitates 3-methyl-cytidine (m3C) methylation at position C32 of the mt-tRNASer(UCN) and mt-tRNAThr. METTL8 knockout cells show a reduction in respiratory chain activity, whereas overexpression increases activity. In pancreatic cancer, METTL8 levels are high, which correlates with lower patient survival and an enhanced respiratory chain activity. Mitochondrial ribosome profiling uncovered mitoribosome stalling on mt-tRNASer(UCN)- and mt-tRNAThr-dependent codons. Further analysis of the respiratory chain complexes using mass spectrometry revealed reduced incorporation of the mitochondrially encoded proteins ND6 and ND1 into complex I. The well-balanced translation of mt-tRNASer(UCN)- and mt-tRNAThr-dependent codons through METTL8-mediated m3C32 methylation might, therefore, facilitate the optimal composition and function of the mitochondrial respiratory chain.

Arctic summer sea ice has decreased dramatically over the last few decades, with a substantial part of this decline attributed to internal variability. However, models show large differences in the...

Expression of human mitochondrial DNA is indispensable for proper function of the oxidative phosphorylation machinery. The mitochondrial genome encodes 22 tRNAs, 2 rRNAs and 11 mRNAs and their post-transcriptional modification constitutes one of the key regulatory steps during mitochondrial gene expression. Cytosine-5 methylation (m5C) has been detected in mitochondrial transcriptome, however its biogenesis has not been investigated in details. Mammalian NOP2/Sun RNA Methyltransferase Family Member 2 (NSUN2) has been characterized as an RNA methyltransferase introducing m5C in nuclear-encoded tRNAs, mRNAs and microRNAs and associated with cell proliferation and differentiation, with pathogenic variants in NSUN2 being linked to neurodevelopmental disorders. Here we employ spatially restricted proximity labelling and immunodetection to demonstrate that NSUN2 is imported into the matrix of mammalian mitochondria. Using three genetic models for NSUN2 inactivation—knockout mice, patient-derived fibroblasts and CRISPR/Cas9 knockout in human cells—we show that NSUN2 is necessary for the generation of m5C at positions 48, 49 and 50 of several mammalian mitochondrial tRNAs. Finally, we show that inactivation of NSUN2 does not have a profound effect on mitochondrial tRNA stability and oxidative phosphorylation in differentiated cells. We discuss the importance of the newly discovered function of NSUN2 in the context of human disease.

The differences between a center-mounted and a side-mounted injector for gasoline direct injection (GDI) applications are analyzed through computational fluid dynamics (CFD). The Engine Combustion Network’s (ECN) axisymmetric 8-hole Spray G injector is compared to a 6-hole injector designed to be side-mounted in an engine. Nozzle-flow simulations are carried out with the commercial CFD software CONVERGE, injecting Euro 5 certification gasoline into a constant volume chamber. Low-load operating conditions are targeted, setting the injection pressure at 50 bar and the ambient pressure to be representative of very early pilot injections. The phase change is handled with the Homogeneous Relaxation Model (HRM), which is assessed and adapted to gasoline flash-boiling conditions. The simulation domains are generated leveraging real injector internal geometries obtained by micron-resolution X-ray tomographic measurements, which introduce manufacturing tolerances and surface roughness in the computational study. Steady needle lift conditions are analyzed. The near-field fuel density distributions and plume morphologies are evaluated, validated and compared to X-ray radiography measurements. A computational best practice is defined and single plume characteristics and variability trends are highlighted as functions of the geometry of the orifices. The plume-plume interaction dynamics are identified and assessed, underlining differences from center- to side-mounted injectors at strong flashing conditions. The obtained numerical framework allows the identification of near-nozzle injection characteristics such as single plume direction, cone angle, spray initial velocity and spatial fuel density distribution. The presented results represent a unique dataset for the initialization of more-affordable Lagrangian spray models, which differentiate the behavior of side-mounted and center-mounted injectors.

Reversible infantile respiratory chain deficiency (RIRCD) is a rare mitochondrial myopathy leading to severe metabolic disturbances in infants, which recover spontaneously after 6‐months of age. RIRCD is associated with the homoplasmic m.14674T>C mitochondrial DNA mutation; however, only ~ 1/100 carriers develop the disease. We studied 27 affected and 15 unaffected individuals from 19 families and found additional heterozygous mutations in nuclear genes interacting with mt‐tRNAGlu including EARS2 and TRMU in the majority of affected individuals, but not in healthy carriers of m.14674T>C, supporting a digenic inheritance. Our transcriptomic and proteomic analysis of patient muscle suggests a stepwise mechanism where first, the integrated stress response associated with increased FGF21 and GDF15 expression enhances the metabolism modulated by serine biosynthesis, one carbon metabolism, TCA lipid oxidation and amino acid availability, while in the second step mTOR activation leads to increased mitochondrial biogenesis. Our data suggest that the spontaneous recovery in infants with digenic mutations may be modulated by the above described changes. Similar mechanisms may explain the variable penetrance and tissue specificity of other mtDNA mutations and highlight the potential role of amino acids in improving mitochondrial disease., Heterozygous mutations in nuclear genes interacting with mt‐tRNAGlu induce the integrated stress response and metabolic rearrangements, reducing penetrance and promoting spontaneous recovery in a rare mitochondrial myopathy.

The multiphase flow inside a diesel injection nozzle is imaged using synchrotron X-rays from the Advanced Photon Source at Argonne National Laboratory. Through acquisitions performed at several viewing angles and subsequent tomographic reconstruction, in-situ 3D visualization is achieved for the first time inside a steel injector at engine-like operating conditions. The morphology of the internal flow reveals strong flow separation and vapor-filled cavities (cavitation), the degree of which correlates with the nozzle’s asymmetric inlet corner profile. Micron-scale surface features, which are artifacts of manufacturing, are shown to influence the morphology of the resulting liquid-gas interface. The data obtained at 0.1 ms time resolution exposes transient flow features and the flow development timescales are shown to be correlated with in-situ imaging of the fuel injector’s hydraulically-actuated valve (needle). As more than 98.5% of the X-ray photon flux is attenuated within the steel injector body itself, we are posed with a unique challenge for imaging the flow within. Time-resolved imaging under these low-light conditions is achieved by exploiting both the refractive and absorptive properties of X-ray photons. The data-processing strategy converted these images with a signal-to-noise ratio of ~ 10 into a meaningful dataset for understanding internal flow and cavitation in a nozzle of diameter 200 μm enclosed within 1–2 millimeters of steel.

X-ray Computed Tomography has gained popularity as a metrology technique for components with detailed internal features. However achieving micron-scale resolution using X-ray CT is challenging. Synchrotron X-ray sources are highly collimated and brilliant, allowing high resolution tomography in metallic components. The X-ray Fuel Spray research at Argonne National Lab is aimed at utilizing synchrotron X-ray diagnostics for providing insights into automotive fuel injection. We present a case study of micro-CT for automotive fuel injectors, with orifices smaller than 100 micrometers. These orifices are imaged with 1 micrometer voxel size with minimal resolvable features of 2 micrometer. Tomographic analysis on large datasets must preserve resolution while being computationally efficient, which is facilitated by deep learning techniques for segmentation developed in-house. The accuracy of segmentation is evaluated using synthetic data. As results, we show high-quality iso-surface extraction and measurements of orifice features for 7 identical fuel injectors, indicating the extent of manufacturing variability. The capabilities developed in our team have potential applicability in the field of metal additive manufacturing, specifically for millimeter-sized components with micron-scale features.

Reversible infantile respiratory chain deficiency (RIRCD) is a rare mitochondrial myopathy leading to severe metabolic disturbances in infants, which recover spontaneously after 6 months of age. RIRCD is associated with the homoplasmic m.14674T>C mitochondrial DNA mutation, however only ∼1/100 carriers develop the disease. We studied 27 affected and 15 unaffected individuals from 19 families and found additional heterozygous mutations in nuclear genes interacting with mt-tRNAGluincludingEARS2andTRMUin the majority of affected individuals, but not in healthy carriers of m.14674T>C, supporting a digenic inheritance. The spontaneous recovery in infants with digenic mutations is modulated by changes in amino acid availability in a multi-step process. First, the integrated stress-response associated with increasedFGF21andGDF15expression enhances catabolism via β-oxidation and the TCA cycle increasing the availability of amino acids. In the second phase mitochondrial biogenesis increases via mTOR activation, leading to improved mitochondrial translation and recovery. Similar mechanisms may explain the variable penetrance and tissue specificity of other mtDNA mutations and highlight the potential role of amino acids in improving mitochondrial disease.