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Marine macroalgae are a key primary producer in coastal ecosystems, but are often overlooked in blue carbon inventories. Large quantities of macroalgal detritus deposit on beaches, but the fate of wrack carbon (C) is little understood. If most of the wrack carbon is respired back to CO2, there would be no net carbon sequestration. However, if most of the wrack carbon is converted to bicarbonate (alkalinity) or refractory DOC, wrack deposition would represent net carbon sequestration if at least part of the metabolic products (e.g., reduced Fe and S) are permanently removed (i.e., long-term burial) and the DOC is not remineralised. To investigate the release of macroalgal C via porewater and its potential to contribute to C sequestration (blue carbon), we monitored the degradation of Ecklonia radiata in flow-through mesocosms simulating tidal flushing on sandy beaches. Over 60 days, 81% of added E. radiata organic matter (OM) decomposed. Per 1 mol of detritus C, the degradation produced 0.48 ± 0.34 mol C of dissolved organic carbon (DOC) (59%) and 0.25 ± 0.07 mol C of dissolved inorganic carbon (DIC) (31%) in porewater, and a small amount of CO2 (0.3 ± 0.0 mol C; ca. 3%) which was emitted to the atmosphere. A significant amount of carbonate alkalinity was found in porewater, equating to 33% (0.27 ± 0.05 mol C) of the total degraded C. The degradation occurred in two phases. In the first phase (days 0–3), 27% of the OM degraded, releasing highly reactive DOC. In the second phase (days 4–60), the labile DOC was converted to DIC. The mechanisms underlying E. radiata degradation were sulphate reduction and ammonification. It is likely that the carbonate alkalinity was primarily produced through sulphate reduction. The formation of carbonate alkalinity and semi-labile or refractory DOC from beach wrack has the potential to play an overlooked role in coastal carbon cycling and contribute to marine carbon sequestration. Graphical abstract

“They call it life, we call it pollution” is an infamous quote which ignores many facts about why carbon dioxide (CO2) poses a significant problem for the ocean. But before we get to this, let’s start at the beginning. All organisms on Earth require a particular set of elements for growth. In the case of plants, these elements are needed to synthesise organic matter in a process called primary production via photosynthesis, and in the case of animals, these elements are directly assimilated by either consuming plant material or by preying on other animals. In this respect, one of the key elements is carbon. Being the molecular backbone for a number of vital organic compounds such as sugars, proteins and nucleic acids (containing genetic information), carbon can be considered as the building block of life.

Climate change is expected to alter the intensity and frequency of upwelling in high productive coastal regions, thus impacting nutrient fluxes, primary productivity and consequently carbon cycling. However, it is unknown how these changes will impact the planktonic (phytoplankton and bacteria) community structure, which affects community respiration (CR) and hence the carbon available for sequestration or transfer to upper trophic levels. Here we present results from a 37-day mesocosm experiment where we examined the response of CR to nutrient additions by simulating upwelling events at different intensities (low, medium, high and extreme) and modes (singular and recurring additions). We also analysed the potential contribution of different plankton size classes and functional groups to CR. The trend in accumulated CR with respect to nutrient fertilisation (total nitrogen added during the experiment) was linear in the two modes. Microplankton (mostly diatoms) and nanoplankton (small flagellates) dominated under extreme upwelling intensities and high CR in both singular and recurring upwelling modes, explaining >65% of the observed variability in CR. In contrast, prokaryotic picoplankton (heterotrophic bacteria and autotrophic cyanobacteria) explained

To ensure dosage compensation for X-linked genes between the sexes, one X chromosome is silenced during early embryonic development of female mammals. This process of X-chromosome inactivation (XCI) is initiated through upregulation of the RNA Xist from one X chromosome shortly after fertilization. Xist then mediates chromosome-wide gene silencing in cis and remains expressed in all cell types except the germ line and the pluripotent state, where XCI is reversed. The factors that drive Xist upregulation and thereby initiate XCI remain however unknown. We identify GATA transcription factors as potent Xist activators and demonstrate that they are essential for the activation of Xist in mice following fertilization. Through a pooled CRISPR activation screen we find that GATA1 can drive ectopic Xist expression in murine embryonic stem cells (mESCs). We demonstrate that all GATA factors can activate Xist directly via a GATA-responsive regulatory element (RE79) positioned 100 kb upstream of the Xist promoter. Additionally, GATA factors are essential for the induction of XCI in mouse preimplantation embryos, as simultaneous deletion of three members of the GATA family (GATA1/4/6) in mouse zygotes effectively prevents Xist upregulation. Thus, initiation of XCI and possibly its maintenance in distinct lineages of the preimplantation embryo is ensured by the combined activity of different GATA family members, and the absence of GATA factors in the pluripotent state likely contributes to X reactivation. We thus describe a form of regulation in which the combined action of numerous tissue-specific factors can achieve near-ubiquitous expression of a target gene.

Earth’s Radiation Budget is partly dictated by the fragile and complex balance between biogenic volatile organic compounds (BVOCs) and greenhouse gases (GHGs), which have the potential to impose cooling or warming once emitted to the atmosphere. Whilst methane (CH4) is strictly associated with global warming due to its solar-radiation absorbing properties, dimethyl sulfide (DMS) is generally considered a cooling gas through the light scattering properties of its atmospheric oxidation products. However, DMS may also partially contribute to the Earth’s warming through a small portion of it being degraded to CH4 in the water column. Coral reefs emit both DMS and CH4 but they have not previously been simultaneously measured. Here, we report DMS and CH4 fluxes as well as aerosol particle counts at Heron Island, southern Great Barrier Reef, during the austral summer of 2016. Sea-to-air DMS and CH4 fluxes were on average 24.9 ± 1.81 and 1.36 ± 0.11 µmol m-2 d-1, whilst intermediate (< 0.5-2.5 um) and large (> 2.5 um) particle number concentrations averaged 5.51 x 106 ± 1.73 x 105 m-3 and 1.15 x 106 ± 4.63 x 104 m-3, respectively. Positive correlations were found between DMS emissions and the abundance of intermediate (R2 = 0.1669, p < 0.001, n = 93) and large (R2 = 0.0869, p = 0.004, n = 93) aerosol particles, suggesting that DMS sea-to-air emissions significantly contribute to the growth of existing particles to the measured size ranges at the Heron Island lagoon. Additionally, a strong positive correlation was found between DMS and CH4 fluxes (R2 = 0.7526, p < 0.00001, n = 93), suggesting that the emission of these volatile compounds from coral reefs is closely linked. The slope of the regression between DMS and CH4 suggests that CH4 emissions at the Heron Island lagoon represent 5% of that of DMS, which is consistent with the average sea-to-air fluxes reported in this study (i.e. 24.9 ± 1.81 µmol m-2 d-1 for DMS and 1.36 ± 0.11 for CH4). These findings provide new insights on the complexity of BVOC and GHG emissions in coral reef systems and their potential role in climate regulation.

One snapshot of the peer-review process for "Somatic XIST activation and features of X chromosome inactivation in male human cancers" (Sadagopan et al., 2022).

Oxygen minimum zones (OMZs) are characterized by enhanced carbon dioxide (CO2) levels and low pH and are being further acidified by uptake of anthropogenic atmospheric CO2. With ongoing intensification and expansion of OMZs due to global warming, carbonate chemistry conditions may become more variable and extreme, particularly in the eastern boundary upwelling systems. In austral summer (February–April) 2017, a large-scale mesocosm experiment was conducted in the coastal upwelling area off Callao (Peru) to investigate the impacts of ongoing ocean deoxygenation on biogeochemical processes, coinciding with a rare coastal El Niño event. Here we report on the temporal dynamics of carbonate chemistry in the mesocosms and surrounding Pacific waters over a continuous period of 50 d with high-temporal-resolution observations (every second day). The mesocosm experiment simulated an upwelling event in the mesocosms by addition of nitrogen (N)-deficient and CO2-enriched OMZ water. Surface water in the mesocosms was acidified by the OMZ water addition, with pHT lowered by 0.1–0.2 and pCO2 elevated to above 900 µatm. Thereafter, surface pCO2 quickly dropped to near or below the atmospheric level (405.22 µatm in 2017; Dlugokencky and Tans, 2021; NOAA/Global Monitoring Laboratory (GML)) mainly due to enhanced phytoplankton production with rapid CO2 consumption. Further observations revealed that the dominance of the dinoflagellate Akashiwo sanguinea and contamination of bird excrements played important roles in the dynamics of carbonate chemistry in the mesocosms. Compared to the simulated upwelling, natural upwelling events in the surrounding Pacific waters occurred more frequently with sea-to-air CO2 fluxes of 4.2–14.0 mmol C m−2 d−1. The positive CO2 fluxes indicated our site was a local CO2 source during our study, which may have been impacted by the coastal El Niño. However, our observations of dissolved inorganic carbon (DIC) drawdown in the mesocosms suggest that CO2 fluxes to the atmosphere can be largely dampened by biological processes. Overall, our study characterized carbonate chemistry in nearshore Pacific waters that are rarely sampled in such a temporal resolution and hence provided unique insights into the CO2 dynamics during a rare coastal El Niño event.

Nuclear Overhauser Effect (NOE) methods in NMR are an important tool for 3D structural analysis of small molecules. Quantitative NOE methods conventionally rely on reference distances, known distances that have to be spectrally separated and are not always available. Here we present a new method for evaluation and 3D structure selection that does not require a reference distance, instead utilizing structures optimized by molecular mechanics, enabling NOE evaluation even on molecules without suitable reference groups., A quantitative Nuclear Overhauser Effect (NOE) analysis approach that avoids the use of and internal reference distance to perform molecular configuration selection.

BACKGROUND Numerous studies investigated cell-based therapies for myocardial infarction (MI). The conflicting results of these studies have established the need for developing innovative approaches for applying cell-based therapy for MI. Experimental studies on animal models demonstrated the potential of fresh, uncultured, unmodified, autologous adipose-derived regenerative cells (UA-ADRCs) for treating acute MI. In contrast, studies on the treatment of chronic MI (CMI; > 4 wk post-MI) with UA-ADRCs have not been published so far. Among several methods for delivering cells to the myocardium, retrograde delivery into a temporarily blocked coronary vein has recently been demonstrated as an effective option. AIM To test the hypothesis that in experimentally-induced chronic myocardial infarction (CMI; > 4 wk post-MI) in pigs, retrograde delivery of fresh, uncultured, unmodified, autologous adipose-derived regenerative cells (UA-ADRCs) into a temporarily blocked coronary vein improves cardiac function and structure. METHODS The left anterior descending (LAD) coronary artery of pigs was blocked for 180 min at time point T0. Then, either 18 × 106 UA-ADRCs prepared at "point of care" or saline as control were retrogradely delivered via an over-the-wire balloon catheter placed in the temporarily blocked LAD vein 4 wk after T0 (T1). Effects of cells or saline were assessed by cardiac magnetic resonance (CMR) imaging, late gadolinium enhancement CMR imaging, and post mortem histologic analysis 10 wk after T0 (T2). RESULTS Unlike the delivery of saline, delivery of UA-ADRCs demonstrated statistically significant improvements in cardiac function and structure at T2 compared to T1 (all values given as mean ± SE): Increased mean LVEF (UA-ADRCs group: 34.3% ± 2.9% at T1 vs 40.4 ± 2.6% at T2, P = 0.037; saline group: 37.8% ± 2.6% at T1 vs 36.2% ± 2.4% at T2, P > 0.999), increased mean cardiac output (UA-ADRCs group: 2.7 ± 0.2 L/min at T1 vs 3.8 ± 0.2 L/min at T2, P = 0.002; saline group: 3.4 ± 0.3 L/min at T1 vs 3.6 ± 0.3 L/min at T2, P = 0.798), increased mean mass of the left ventricle (UA-ADRCs group: 55.3 ± 5.0 g at T1 vs 71.3 ± 4.5 g at T2, P < 0.001; saline group: 63.2 ± 3.4 g at T1 vs 68.4 ± 4.0 g at T2, P = 0.321) and reduced mean relative amount of scar volume of the left ventricular wall (UA-ADRCs group: 20.9% ± 2.3% at T1 vs 16.6% ± 1.2% at T2, P = 0.042; saline group: 17.6% ± 1.4% at T1 vs 22.7% ± 1.8% at T2, P = 0.022). CONCLUSION Retrograde cell delivery of UA-ADRCs in a porcine model for the study of CMI significantly improved myocardial function, increased myocardial mass and reduced the formation of scar tissue.

Diatoms, large bloom-forming marine microorganisms, build frustules out of silicate, which ballasts the cells and aids their export to the deep ocean. This unique physiology forges an important link between the marine silicon and carbon cycles. However, the effect of ocean acidification on the silicification of diatoms is unclear. Here we show that diatom silicification strongly diminishes with increased acidity in a natural Antarctic community. Analyses of single cells from within the community reveal that the effect of reduced pH on silicification differs among taxa, with several species having significantly reduced silica incorporation at CO2 levels equivalent to those projected for 2100. These findings suggest that, before the end of this century, ocean acidification may influence the carbon and silicon cycle by both altering the composition of the diatom assemblages and reducing cell ballasting, which will probably alter vertical flux of these elements to the deep ocean. Diatoms have silicate skeletons that affect their buoyancy in the ocean. Ocean acidification reduces silicification, with varying effects between species, and could alter the marine carbon and silica cycles through changes in community composition and sinking rates.

Objective assessment of surgical skill is gaining traction in a number of specialty fields. In robot-assisted surgery in particular, the availability of data from the operating console and patient-side robot offers the potential to derive objective metrics of performance based on tool movement kinematics. While these techniques are becoming established in the laparoscopic domain, current assessment techniques for robotic endovascular surgery are based primarily on observation, checklists, and grading scales. This work presents an objective and quantitative means of measuring technical competence based on analysis of the kinematics of endovascular tool tip motions controlled with a robotic interface. We designed an experiment that recorded catheter tip movement from 21 subjects performing fundamental endovascular robotic navigation tasks on a physical model. Motion-based measures of smoothness (spectral arc length and number of submovements) were computed and tested for correlation with subjective scores from a global rating scale assessment tool that has been validated for use when performing manual catheterization. Results show that the smoothness metrics that produced significant correlations with the global rating scale for manual catheterization show similar correlations for robotic catheterization. This finding is notable, since with the robotic interface, tool tip motion is commanded discretely via a control button interface, while in manual procedures the tools are controlled through continuous movements of the surgeon's hands. Logistic regression analysis using a single motion metric was capable of classifying subjects by expertise with better than 90% accuracy. These objective and quantitative metrics that capture movement quality could be incorporated into future training protocols to provide detailed feedback on trainee performance.

Ocean acidification (OA) can induce shifts in plankton community composition, with coccolithophores being mostly negatively impacted. This is likely to change particulate inorganic and organic carbon (PIC and POC, respectively) production, with impacts on the biological carbon pump. Hence, assessing and, most importantly, understanding species‐specific sensitivities of coccolithophores is paramount. In a multispecies comparison, spanning more than two orders of magnitude in terms of POC and PIC production rates, among Calcidiscus leptoporus, Coccolithus pelagicus subsp. braarudii, Emiliania huxleyi, Gephyrocapsa oceanica, and Scyphosphaera apsteinii, we found that cellular PIC : POC was a good predictor for a species’ OA sensitivity. This is likely related to the need for cellular pH homeostasis, which is challenged by the process of calcification producing protons internally, especially when seawater pH decreases in an OA scenario. With higher PIC : POC, species and strains being more sensitive to OA coccolithophores may shift toward less calcified varieties in the future.

SummaryGene-regulatory networks control establishment and maintenance of alternative gene expression states during development. A particular challenge is the acquisition of opposing states by two copies of the same gene, as it is the case in mammals for Xist at the onset of random X-chromosome inactivation (XCI). The regulatory principles that lead to stable mono-allelic expression of Xist remain unknown. Here, we uncovered the minimal Xist regulatory network, by combining mathematical modeling and experimental validation of central model predictions. We identified a symmetric toggle switch as the basis for random mono-allelic Xist up-regulation, which reproduces data from several mutant, aneuploid and polyploid murine cell lines with various Xist expression patterns. Moreover, this toggle switch explains the diversity of strategies employed by different species at the onset of XCI. In addition to providing a unifying conceptual framework to explore X-chromosome inactivation across mammals, our study sets the stage for identifying the molecular mechanisms required to initiate random XCI.

Upwelling of nutrient-rich deep waters make eastern boundary upwelling systems (EBUSs), such as the Humboldt Current system, hot spots of marine productivity. Associated settling of organic matter to depth and consecutive aerobic decomposition results in large subsurface water volumes being oxygen depleted. Under these circumstances, organic matter remineralisation can continue via denitrification, which represents a major loss pathway for bioavailable nitrogen. Additionally, anaerobic ammonium oxidation can remove significant amounts of nitrogen in these areas. Here we assess the interplay of suboxic water upwelling and nitrogen cycling in a manipulative offshore mesocosm experiment. Measured denitrification rates in incubations with water from the oxygen-depleted bottom layer of the mesocosms (via 15N label incubations) mostly ranged between 5.5 and 20 nmol N2 L−1 h−1 (interquartile range), reaching up to 80 nmol N2 L−1 h−1. However, actual in situ rates in the mesocosms, estimated via Michaelis–Menten kinetic scaling, did most likely not exceed 0.2–4.2 nmol N2 L−1 h−1 (interquartile range) due to substrate limitation. In the surrounding Pacific, measured denitrification rates were similar, although indications of substrate limitation were detected only once. In contrast, anammox (anaerobic ammonium oxidation) made only a minor contribution to the overall nitrogen loss when encountered in both the mesocosms and the Pacific Ocean. This was potentially related to organic matter C / N stoichiometry and/or process-specific oxygen and hydrogen sulfide sensitivities. Over the first 38 d of the experiment, total nitrogen loss calculated from in situ rates of denitrification and anammox was comparable to estimates from a full nitrogen budget in the mesocosms and ranged between ∼ 1 and 5.5 µmol N L−1. This represents up to ∼ 20 % of the initially bioavailable inorganic and organic nitrogen standing stocks. Interestingly, this loss is comparable to the total amount of particulate organic nitrogen that was exported into the sediment traps at the bottom of the mesocosms at about 20 m depth. Altogether, this suggests that a significant portion, if not the majority of nitrogen that could be exported to depth, is already lost, i.e. converted to N2 in a relatively shallow layer of the surface ocean, provided that there are oxygen-deficient conditions like those during coastal upwelling in our study. Published data for primary productivity and nitrogen loss in all EBUSs reinforce such conclusion.

Primary production in the Southern Ocean is dominated by diatom-rich phytoplankton assemblages, whose individual physiological characteristics and community composition are strongly shaped by the environment, yet knowledge on how diatoms allocate cellular energy in response to ocean acidification (OA) is limited. Understanding such changes in allocation is integral to determining the nutritional quality of diatoms and the subsequent impacts on the trophic transfer of energy and nutrients. Using synchrotron-based Fourier transform infrared microspectroscopy, we analysed the macromolecular content of selected individual diatom taxa from a natural Antarctic phytoplankton community exposed to a gradient of fCO2 levels (288-1263 µatm). Strong species-specific differences in macromolecular partitioning were observed under OA. Large taxa showed preferential energy allocation towards proteins, while smaller taxa increased both lipid and protein stores at high fCO2 . If these changes are representative of future Antarctic diatom physiology, we may expect a shift away from lipid-rich large diatoms towards a community dominated by smaller taxa, but with higher lipid and protein stores than their present-day contemporaries, a response that could have cascading effects on food web dynamics in the Antarctic marine ecosystem.

Developmental genes such asXist, the master regulator of X-chromosome inactivation (XCI), are controlled by complexcis-regulatory landscapes, which decode multiple signals to establish specific spatio-temporal expression patterns.Xistintegrates information on X-chromosomal dosage and developmental stage to trigger XCI at the primed pluripotent state in females only. Through a pooled CRISPR interference screen in differentiating mouse embryonic stem cells, we identify functional enhancer elements ofXistduring the onset of random XCI. By quantifying how enhancer activity is modulated by X-dosage and differentiation, we find that X-dosage controls the promoter-proximal region in a binary switch-like manner. By contrast, differentiation cues activate a series of distal elements and bring them into closer spatial proximity of theXistpromoter. The strongest distal element is part of an enhancer cluster ∼200 kb upstream of theXistgene which is associated with a previously unannotatedXist-enhancing regulatory transcript, we namedXert. Developmental cues and X-dosage are thus decoded by distinct regulatory regions, which cooperate to ensure female-specificXistupregulation at the correct developmental time. Our study is the first step to disentangle how multiple, functionally distinct regulatory regions interact to generate complex expression patterns in mammals.

BackgroundX-chromosomal genes contribute to sex differences, in particular during early development, when both X chromosomes are active in females. Double X-dosage shifts female pluripotent cells towards the naive stem cell state by increasing pluripotency factor expression, inhibiting the differentiation-promoting MAP kinase (MAPK) signaling pathway, and delaying differentiation.ResultsTo identify the genetic basis of these sex differences, we use a two-step CRISPR screening approach to comprehensively identify X-linked genes that cause the female pluripotency phenotype in murine embryonic stem cells. A primary chromosome-wide CRISPR knockout screen and three secondary screens assaying for different aspects of the female pluripotency phenotype allow us to uncover multiple genes that act in concert and to disentangle their relative roles. Among them, we identify Dusp9 and Klhl13 as two central players. While Dusp9 mainly affects MAPK pathway intermediates, Klhl13 promotes pluripotency factor expression and delays differentiation, with both factors jointly repressing MAPK target gene expression.ConclusionsHere, we elucidate the mechanisms that drive sex-induced differences in pluripotent cells and our approach serves as a blueprint to discover the genetic basis of the phenotypic consequences of other chromosomal effects.

Upwelling of nutrient-rich deep waters make Eastern Boundary upwelling systems (EBUS), such as the Humboldt Current System, hotspots of marine productivity. Associated settling of organic matter to depth and consecutive aerobic de composition results in large sub-surface water volumes being oxygen-depleted. Under these circumstances organic matter remineralisation can continue via denitrification which represents a major loss pathway of bioavailable nitrogen. Another process removing significant amounts of nitrogen in these areas is anaerobic ammonium oxidation. Here we assess the interplay of suboxic water upwelling and nitrogen cycling in a manipulative off-shore mesocosm experiment. Measured denitrification rates in the oxygen-depleted bottom layer of the mesocosms mostly ranged between 5.5–20 (interquartile range), reaching up to 80 nmol N2 L−1 h−1. However, realised in-situ rates did most likely not exceed 0.6–1.6 nmol N2 L−1 h−1 (interquartile range), due to substrate limitation in the mesocosms. This was in contrast to realised rates in the surrounding Pacific. Both in the mesocosms and the Pacific Ocean anammox made only a minor contribution to overall nitrogen loss when encountered. Over the first 38 days of the experiment, total nitrogen loss calculated from denitrification and anammox rates was comparable to estimates from a full nitrogen budget in the mesocosms and ranged between ∼ 2–6 μmol N L−1. This represents up to ∼ 20 % of the initially bioavailable inorganic and organic nitrogen standing stocks. Interestingly, this loss is comparable to the total amount of particulate organic nitrogen that was exported into the sediment traps at the bottom of the mesocosms in about 20 metres depth. Altogether, this suggests that a significant portion, if not the majority of nitrogen that could be exported to depth, is already lost, i.e. converted to N2 in a relatively shallow layer of the surface ocean, provided oxygen-deficient conditions like during coastal upwelling in our study.

Developmental genes such as Xist, which initiates X chromosome inactivation, are controlled by complex cis-regulatory landscapes, which decode multiple signals to establish specific spatiotemporal expression patterns. Xist integrates information on X chromosome dosage and developmental stage to trigger X inactivation in the epiblast specifically in female embryos. Through a pooled CRISPR screen in differentiating mouse embryonic stem cells, we identify functional enhancer elements of Xist at the onset of random X inactivation. Chromatin profiling reveals that X-dosage controls the promoter-proximal region, while differentiation cues activate several distal enhancers. The strongest distal element lies in an enhancer cluster associated with a previously unannotated Xist-enhancing regulatory transcript, which we named Xert. Developmental cues and X-dosage are thus decoded by distinct regulatory regions, which cooperate to ensure female-specific Xist upregulation at the correct developmental time. With this study, we start to disentangle how multiple, functionally distinct regulatory elements interact to generate complex expression patterns in mammals.

Pathognomonic PAX3-FOXO1 fusion oncogene expression is associated with poor outcome in rhabdomyosarcoma. Combining genome-wide CRISPR screening with cell- based functional genetic approaches, we here provide evidence that PAX3-FOXO1 induces replication stress, resulting in a synthetic lethal dependency to ATR-mediated DNA damage-response signaling in rhabdomyosarcoma. Expression of PAX3-FOXO1 in muscle progenitor cells was not only sufficient to induce hypersensitivity to ATR inhibition, but PAX3-FOXO1-expressing rhabdomyosarcoma cells also exhibited increased sensitivity to structurally diverse inhibitors of ATR, a dependency that could be validated genetically. Mechanistically, ATR inhibition led to replication stress exacerbation, decreased BRCA1 phosphorylation and reduced homologous recombination-mediated DNA repair pathway activity. Consequently, ATR inhibitor treatment increased sensitivity of rhabdomyosarcoma cells to PARP inhibitionin vitro, and combined ATR and PARP inhibition induced regression of primary patient-derived alveolar rhabdomyosarcoma xenograftsin vivo. Moreover, a genome-wide CRISPR activation screen (CRISPRa) identifiedFOSgene family members as inducers of resistance against ATR inhibitors. Mechanistically,FOSgene family members reduced replication stress in rhabdomyosarcoma cells. Lastly, compassionate use of ATR inhibitors in two pediatric patients suffering from relapsed PAX3-FOXO1-expressing alveolar rhabdomyosarcoma showed signs of tolerability, paving the way to clinically exploit this novel synthetic lethal dependency in rhabdomyosarcoma.

Large-scale dinoflagellate blooms have appeared in recent decades in the Taiwan Strait, Southeast China. To study spatial variability of phytoplankton community composition, physical and chemical environmental drivers in surface seawater of the Taiwan Strait, we conducted cruises in May and July 2019. Cell numbers of dinoflagellates were significantly higher than that of diatoms in most sampling stations during the cruise in May, whereas diatoms were the major contributor to autotrophic biomass in July. Phytoplankton community shifted from a dinoflagellate- and diatom-dominated system in May to diatom dominance in July. The dominant phytoplankton species (genera) were the harmful algal bloom dinoflagellates Prorocentrum donghaiense and Scrippsiella trochoidea and the diatoms Coscinodiscus in May, and Rhizosolenia, Pseudo-nitzschia, and Guinardia in July. Cell densities of dinoflagellates and P. donghaiense reduced exponentially with increasing seawater temperature and salinity and decreasing dissolved inorganic nitrogen (DIN) concentrations. Based on the results of our work and previous studies, it becomes obvious that harmful dinoflagellate blooms are likely to be a major component of the planktonic food web in the Taiwan Strait at a temperature of 17.0&ndash, 23.0 &deg, C, a salinity of 29.0&ndash, 33.0 psu, and a DIN concentration higher than 2.0 &mu, mol L&ndash, 1.

Eastern boundary upwelling systems (EBUS) are among the most productive marine ecosystems on Earth. The production of organic material is fueled by upwelling of nutrient-rich deep waters and high incident light at the sea surface. However, biotic and abiotic factors can modify surface production and related biogeochemical processes. Determining these factors is important because EBUS are considered hotspots of climate change, and reliable predictions of their future functioning requires understanding of the mechanisms driving the biogeochemical cycles therein. In this field experiment, we used in situ mesocosms as tools to improve our mechanistic understanding of processes controlling organic matter cycling in the coastal Peruvian upwelling system. Eight mesocosms, each with a volume of ∼55 m3, were deployed for 50 d ∼6 km off Callao (12∘ S) during austral summer 2017, coinciding with a coastal El Niño phase. After mesocosm deployment, we collected subsurface waters at two different locations in the regional oxygen minimum zone (OMZ) and injected these into four mesocosms (mixing ratio ≈1.5 : 1 mesocosm: OMZ water). The focus of this paper is on temporal developments of organic matter production, export, and stoichiometry in the individual mesocosms. The mesocosm phytoplankton communities were initially dominated by diatoms but shifted towards a pronounced dominance of the mixotrophic dinoflagellate (Akashiwo sanguinea) when inorganic nitrogen was exhausted in surface layers. The community shift coincided with a short-term increase in production during the A. sanguinea bloom, which left a pronounced imprint on organic matter C : N : P stoichiometry. However, C, N, and P export fluxes did not increase because A. sanguinea persisted in the water column and did not sink out during the experiment. Accordingly, export fluxes during the study were decoupled from surface production and sustained by the remaining plankton community. Overall, biogeochemical pools and fluxes were surprisingly constant for most of the experiment. We explain this constancy by light limitation through self-shading by phytoplankton and by inorganic nitrogen limitation which constrained phytoplankton growth. Thus, gain and loss processes remained balanced and there were few opportunities for blooms, which represents an event where the system becomes unbalanced. Overall, our mesocosm study revealed some key links between ecological and biogeochemical processes for one of the most economically important regions in the oceans.

Relative to their surface area, estuaries make a disproportionately large contribution of dissolved organic carbon (DOC) to the global carbon cycle, but it is unknown how this will change under a future climate. As such, the response of DOC fluxes from microbially dominated unvegetated sediments to individual and combined future climate stressors of temperature change (from Δ−3 to Δ+5 ∘C compared to ambient mean temperatures) and ocean acidification (OA, ∼ 2× current CO2 partial pressure, pCO2) was investigated ex situ. Warming alone increased sediment heterotrophy, resulting in a proportional increase in sediment DOC uptake; sediments became net sinks of DOC (3.5 to 8.8 mmol C m−2 d−1) at warmer temperatures (Δ+3 and Δ+5 ∘C, respectively). This temperature response changed under OA conditions, with sediments becoming more autotrophic and a greater sink of DOC (up to 4× greater than under current pCO2 conditions). This response was attributed to the stimulation of heterotrophic bacteria with the autochthonous production of labile organic matter by microphytobenthos. Extrapolating these results to the global area of unvegetated subtidal estuarine sediments, we find that the future climate of warming (Δ+3 ∘C) and OA may decrease estuarine export of DOC by ∼ 80 % (∼ 150 Tg C yr−1) and have a disproportionately large impact on the global DOC budget.

Patients with small vessel cerebrovascular disease frequently suffer from apathy, a debilitating neuropsychiatric syndrome, the underlying mechanisms of which remain to be established. Here we investigated the hypothesis that apathy is associated with disrupted decision making in effort-based decision making, and that these alterations are associated with abnormalities in the white matter network connecting brain regions that underpin such decisions. Eighty-two patients with MRI evidence of small vessel disease were assessed using a behavioural paradigm as well as diffusion weighted MRI. The decision-making task involved accepting or rejecting monetary rewards in return for performing different levels of physical effort (hand grip force). Choice data and reaction times were integrated into a drift diffusion model that framed decisions to accept or reject offers as stochastic processes approaching a decision boundary with a particular drift rate. Tract-based spatial statistics were used to assess the relationship between white matter tract integrity and apathy, while accounting for depression. Overall, patients with apathy accepted significantly fewer offers on this decision-making task. Notably, while apathetic patients were less responsive to low rewards, they were also significantly averse to investing in high effort. Significant reductions in white matter integrity were observed to be specifically related to apathy, but not to depression. These included pathways connecting brain regions previously implicated in effort-based decision making in healthy people. The drift rate to decision parameter was significantly associated with both apathy and altered white matter tracts, suggesting that both brain and behavioural changes in apathy are associated with this single parameter. On the other hand, depression was associated with an increase in the decision boundary, consistent with an increase in the amount of evidence required prior to making a decision. These findings demonstrate altered effort-based decision making for reward in apathy, and also highlight dissociable mechanisms underlying apathy and depression in small vessel disease. They provide clear potential brain and behavioural targets for future therapeutic interventions, as well as modelling parameters that can be used to measure the effects of treatment at the behavioural level., Apathy is common in patients with cerebrovascular small vessel disease. Saleh et al. show that apathy is associated with altered effort-based decision making for reward, and that these changes correlate with abnormalities in the white matter network connecting the medial frontal regions and basal ganglia

High-latitude oceans have been identified as particularly vulnerable to ocean acidification if anthropogenic CO2 emissions continue. Marine microbes are an essential part of the marine food web and are a critical link in biogeochemical processes in the ocean, such as the cycling of nutrients and carbon. Despite this, the response of Antarctic marine microbial communities to ocean acidification is poorly understood. We investigated the effect of increasing fCO2 on the growth of heterotrophic nanoflagellates (HNFs), nano- and picophytoplankton, and prokaryotes (heterotrophic Bacteria and Archaea) in a natural coastal Antarctic marine microbial community from Prydz Bay, East Antarctica. At CO2 levels ≥634 µatm, HNF abundance was reduced, coinciding with increased abundance of picophytoplankton and prokaryotes. This increase in picophytoplankton and prokaryote abundance was likely due to a reduction in top-down control of grazing HNFs. Nanophytoplankton abundance was elevated in the 634 µatm treatment, suggesting that moderate increases in CO2 may stimulate growth. The taxonomic and morphological differences in CO2 tolerance we observed are likely to favour dominance of microbial communities by prokaryotes, nanophytoplankton, and picophytoplankton. Such changes in predator–prey interactions with ocean acidification could have a significant effect on the food web and biogeochemistry in the Southern Ocean, intensifying organic-matter recycling in surface waters; reducing vertical carbon flux; and reducing the quality, quantity, and availability of food for higher trophic levels.

The brain's impotence to utilize long-chain fatty acids as fuel, one of the dogmas in neuroscience, is surprising, since the nervous system is the tissue most energy consuming and most vulnerable to a lack of energy. Challenging this view, we here show in vivo that loss of the Drosophila carnitine palmitoyltransferase 2 (CPT2), an enzyme required for mitochondrial β-oxidation of long-chain fatty acids as substrates for energy production, results in the accumulation of triacylglyceride-filled lipid droplets in adult Drosophila brain but not in obesity. CPT2 rescue in glial cells alone is sufficient to restore triacylglyceride homeostasis, and we suggest that this is mediated by the release of ketone bodies from the rescued glial cells. These results demonstrate that the adult brain is able to catabolize fatty acids for cellular energy production.

To ensure dosage compensation between the sexes, one randomly chosen X chromosome is silenced in each female cell in the process of X-chromosome inactivation (XCI). XCI is initiated during early development through upregulation of the long non-coding RNA Xist, which mediates chromosome-wide gene silencing. Cell differentiation, Xist upregulation and silencing are thought to be coupled at multiple levels to ensure inactivation of exactly one out of two X chromosomes. Here we perform an integrated analysis of all three processes through allele-specific single-cell RNA-sequencing. Specifically, we assess the onset of random XCI with high temporal resolution in differentiating mouse embryonic stem cells, and develop dedicated analysis approaches. By exploiting the inter-cellular heterogeneity of XCI onset, we identify Nanog downregulation as its main trigger and discover additional putative Xist regulators. Moreover, we confirm several predictions of the stochastic model of XCI where monoallelic silencing is thought to be ensured through negative feedback regulation. Finally, we show that genetic variation modulates the XCI process at multiple levels, providing a potential explanation for the long-known Xce effect, which leads to preferential inactivation of a specific X chromosome in inter-strain crosses. We thus draw a detailed picture of the different levels of regulation that govern the initiation of XCI. The experimental and computational strategies we have developed here will allow us to profile random XCI in more physiological contexts, including primary human cells in vivo.

Coccolithophore responses to changes in carbonate chemistry speciation such as CO2 and H+ are highly modulated by light intensity and temperature. Here, we fit an analytical equation, accounting for simultaneous changes in carbonate chemistry speciation, light and temperature, to published and original data for Emiliania huxleyi, and compare the projections with those for Gephyrocapsa oceanica. Based on our analysis, the two most common bloom-forming species in present-day coccolithophore communities appear to be adapted for a similar fundamental light niche but slightly different ones for temperature and CO2, with E. huxleyi having a tolerance to lower temperatures and higher CO2 levels than G. oceanica. Based on growth rates, a dominance of E. huxleyi over G. oceanica is projected below temperatures of 22 ∘C at current atmospheric CO2 levels. This is similar to a global surface sediment compilation of E. huxleyi and G. oceanica coccolith abundances suggesting temperature-dependent dominance shifts. For a future Representative Concentration Pathway (RCP) 8.5 climate change scenario (1000 µatm fCO2), we project a CO2 driven niche contraction for G. oceanica to regions of even higher temperatures. However, the greater sensitivity of G. oceanica to increasing CO2 is partially mitigated by increasing temperatures. Finally, we compare satellite-derived particulate inorganic carbon estimates in the surface ocean with a recently proposed metric for potential coccolithophore success on the community level, i.e. the temperature-, light- and carbonate-chemistry-dependent CaCO3 production potential (CCPP). Based on E. huxleyi alone, as there was interestingly a better correlation than when in combination with G. oceanica, and excluding the Antarctic province from the analysis, we found a good correlation between CCPP and satellite-derived particulate inorganic carbon (PIC) with an R2 of 0.73, p

Although coccolithophore physiological responses to CO2-induced changes in seawater carbonate chemistry have been widely studied in the past, there is limited knowledge on the variability of physiological responses between populations from different areas. In the present study, we investigated the specific responses of growth, particulate organic (POC) and inorganic carbon (PIC) production rates of three populations of the coccolithophore Emiliania huxleyi from three regions in the North Atlantic Ocean (Azores: six strains, Canary Islands: five strains, and Norwegian coast near Bergen: six strains) to a CO2 partial pressure (pCO2) range from 120 to 2630 µatm. Physiological rates of each population and individual strain increased with rising pCO2 levels, reached a maximum and declined thereafter. Optimal pCO2 for growth, POC production rates, and tolerance to low pH (i.e., high proton concentration) was significantly higher in an E. huxleyi population isolated from the Norwegian coast than in those isolated near the Azores and Canary Islands. This may be due to the large environmental variability including large pCO2 and pH fluctuations in coastal waters off Bergen compared to the rather stable oceanic conditions at the other two sites. Maximum growth and POC production rates of the Azores and Bergen populations were similar and significantly higher than that of the Canary Islands population. This pattern could be driven by temperature–CO2 interactions where the chosen incubation temperature (16 °C) was slightly below what strains isolated near the Canary Islands normally experience. Our results indicate adaptation of E. huxleyi to their local environmental conditions and the existence of distinct E. huxleyi populations. Within each population, different growth, POC, and PIC production rates at different pCO2 levels indicated strain-specific phenotypic plasticity. Accounting for this variability is important to understand how or whether E. huxleyi might adapt to rising CO2 levels.

Coccolithophore responses to changes in carbonate chemistry speciation such as CO2 and H+ are highly modulated by light intensity and temperature. Here, we fit an analytical equation, accounting for simultaneous changes in carbonate chemistry speciation, light and temperature, to published and original data for Emiliania huxleyi, and compare the projections with those for Gephyrocapsa oceanica. Based on our analysis, the two most common bloom-forming species in present-day coccolithophore communities appear to be adapted for a similar fundamental light niche but slightly different ones for temperature and CO2, with E. huxleyi having a tolerance to lower temperatures and higher CO2 levels than G. oceanica. Based on growth rates, a dominance of E. huxleyi over G. oceanica is projected below temperatures of 22 °C at current atmospheric CO2 levels. This is similar to a global surface sediment compilation of E. huxleyi and G. oceanica coccolith abundances suggesting temperature-dependent dominance shifts. For a future Representative Concentration Pathway (RCP) 8.5 climate change scenario (1000 µatm fCO2), we project a CO2 driven niche contraction for G. oceanica to regions of even higher temperatures. However, the greater sensitivity of G. oceanica to increasing CO2 is partially mitigated by increasing temperatures. Finally, we compare satellite-derived particulate inorganic carbon estimates in the surface ocean with a recently proposed metric for potential coccolithophore success on the community level, i.e. the temperature-, light- and carbonate-chemistry-dependent CaCO3 production potential (CCPP). Based on E. huxleyi alone, as there was interestingly a better correlation than when in combination with G. oceanica, and excluding the Antarctic province from the analysis, we found a good correlation between CCPP and satellite-derived particulate inorganic carbon (PIC) with an R2 of 0.73, p R2 of 0.85, p

Antarctic near-shore waters are amongst the most sensitive in the world to ocean acidification. Microbes occupying these waters are critical drivers of ecosystem productivity, elemental cycling and ocean biogeochemistry, yet little is known about their sensitivity to ocean acidification. A six-level, dose–response experiment was conducted using 650 L incubation tanks (minicosms) adjusted to a gradient in fugacity of carbon dioxide (fCO2) from 343 to 1641 µatm. The six minicosms were filled with near-shore water from Prydz Bay, East Antarctica, and the protistan composition and abundance was determined by microscopy during 18 days of incubation. No CO2-related change in the protistan community composition was observed during the initial 8 day acclimation period under low light. Thereafter, the response of both autotrophic and heterotrophic protists to fCO2 was species-specific. The response of diatoms was mainly cell size related; microplanktonic diatoms (> 20 µm) increased in abundance with low to moderate fCO2 (343–634 µatm) but decreased at fCO2 ≥ 953 µatm. Similarly, the abundance of Phaeocystis antarctica increased with increasing fCO2 peaking at 634 µatm. Above this threshold the abundance of micro-sized diatoms and P. antarctica fell dramatically, and nanoplanktonic diatoms (≤ 20 µm) dominated, therefore culminating in a significant change in the protistan community composition. Comparisons of these results with previous experiments conducted at this site show that the fCO2 thresholds are similar, despite seasonal and interannual differences in the physical and biotic environment. This suggests that near-shore microbial communities are likely to change significantly near the end of this century if anthropogenic CO2 release continues unabated, with profound ramifications for near-shore Antarctic ecosystem food webs and biogeochemical cycling.