dataesr/scientific-paragraphs-categorization huggingface dataset:
This resulted in reduced levels of AMPK abundance/activity (based on phosphorylation of Threonine 172) and increased CCL2 mRNA expression/secretion (Fig. 6a-d ). In contrast, the pan-AMPK activator PF-739 stimulated AMPK activity without affecting the levels of CK-B, and abrogated the induction of CCL2 transcription by tumour necrosis factor alpha (TNF-α), a canonical positive regulator of inflammation (Extended Data Fig. 6a, b )  36, 37. After establishing a causal link between AMPK and CCL2 expression in human white adipocytes, we next measured AMPK phosphorylation in cells transfected with siCKB or incubated with phosphocreatine. Compared to control cells, AMPK phosphorylation was reduced in both conditions (Fig. 6e, f ). To test if the effects of CKB depletion on CCL2 expression depend on reduced AMPK activity, we incubated siCKB-and siC-transfected cells with and without PF-739.
Murine in vivo models corroborate human findings. To assess the relevance of our findings in vivo, we analysed published datasets and investigated different mouse models. First, we mined single-nucleus transcriptomics of epididymal WAT  38  and observed that, similar to humans, Ckb is the most highly expressed creatine kinase-encoding gene in white adipocytes (Extended Data Fig. 7a ). As obese humans display decreased CKB expression and increased phosphocreatine levels, we next determined if a short (five weeks) high fat diet (HFD) regimen recapitulated this phenotype in mice. This intervention length was chosen to determine if alterations in phosphocreatine/creatine metabolism are early events in the development of obesity. As expected, HFD-feeding resulted in increased body weight and fat mass (Extended Data Fig.
7b, c ), impaired glucose tolerance (Extended Data Fig. 7d ) and WAT inflammation with increased gene expression of Ccl2, as well as the macrophage markers Cd68 and Adgre1 (F4/80), where the latter was confirmed by immunofluorescence analysis (Extended Data Fig. 7e, f ). Following this general characterization, we measured CK-B mRNA/protein and phosphocreatine/creatine levels in WAT. We found that mice fed HFD exhibited lower levels of Ckb mRNA (Fig. 7a ) and CK-B protein, where the latter was observed both in total tissue lysates and in intact tissue by immunofluorescence analysis (Fig. 7b, c ). This was accompanied by higher phosphocreatine levels and phosphocreatine/creatine ratio compared to mice on chow diet (CD; Fig. 7d ). In concordance with the findings in our clinical cohorts, these results confirm that the phosphocreatine/creatine pathway is perturbed by weight gain in mice.
To determine if elevating phosphocreatine levels was sufficient to enhance WAT inflammation in vivo, we administered phosphocreatine (3 mg g -1 ) or phosphate buffered saline (PBS) intraperitoneally in chow-fed mice daily for seven days. This short-term treatment resulted in significant increases in WAT phosphocreatine levels (Fig. 7e ) but did not influence body weight, WAT weight, fasting plasma glucose and insulin (Extended Data Fig. 7g-j ) or Ckb mRNA and protein levels (Fig. 7f, g ). However, we observed an increase in the expression of Ccl2 and Cd68 (with a trend for an increase in Adgre1) (Fig. 7f ) and increased immunofluorescence staining of F4/80 following phosphocreatine injections (Fig. 7h ). As intraperitoneal injections with high doses of phosphocreatine may affect multiple organs, we determined the effects of Ckb depletion on WAT inflammation in vivo by comparing adipocyte-specific Ckb deleted (Ckb Adipoq-Cre ) with control (Ckb fl/fl ) mice fed a HFD for 16 weeks.
This results in altered ATP/ADP levels, which in turn attenuate AMPK activity leading to increased transcription of multiple pro-inflammatory genes including the chemokine CCL2. Collectively, our data unravel a link between energy shuttling and inflammation in white adipocytes and suggest that this balance is perturbed in obesity. Fig. 5  | Phosphocreatine is taken up by SLC6A8 in white adipocytes and promotes CCL2 production. a, Human adipocytes were incubated with PBS or different concentrations of phosphocreatine for 24 h and effects on CCL2 mRNA expression were determined (three replicates per condition, repeated twice). Overall P = 0. 0021. b, The ratio of phosphocreatine/creatine in human in vitro differentiated adipocytes incubated with PBS or 30 mmol l -1 phosphocreatine for 24 h (three replicates per condition, repeated twice).
For each section, the total number of F4/80 positive cells were counted in three to four random ×10 fields using a Axio Observer. Z1 inverted fluorescence microscope (Zeiss) and the AxioVision software. Adipocyte progenitors were stained for DPP4. In brief, cells were seeded on glass coverslips (thickness no. 1. 5) and fixed in 4% PFA for 15 min at room temperature. Cells were washed twice with PBS, permeabilized using 0. 1% Triton-X100 for 10 min and blocked for 30 min in PBS containing 10% normal goat serum. Subsequently, cells were incubated for 1 h with an antibody targeting DPP4 in blocking buffer. Cells were washed three times with PBS and incubated with secondary Alexa Fluor-conjugated antibodies for 1 h. Following three washes with PBS, cells were stained with Hoechst (diluted 1:5,000) for 15 min and mounted in fluorescence mounting medium (Fluoromount Aqueous Mounting Medium, refractive index 1.
4 oil objective was used to acquire images. The ×1. 5 tube lens of the Ti2 was inserted to reach Nyquist sampling in xy. A zstack was acquired at Nyquist sampling in z and the stack was deconvolved using Nikon NIS element software and the fast deconvolution algorithm. Mitochondrial content in differentiated adipocytes was transferred to a fresh reaction tube and evaporated to dryness using a centrifugal evaporator. Proteins were extracted from the residue cell debris pellet and quantified using a BCA Kit (Sigma-Aldrich). Metabolite profiling was performed using a Sciex Exion AD LC coupled to a Sciex X500R Q-ToF-MS (Sciex). Articles NATurE METABOLISM Articles NATurE METABOLISM Extended Data Fig.
Acknowledgements We thank  L. Kazak ,  B. Samborska  and  J. Rahbani  ( McGill University, Montreal ) for providing samples from the adipocyte-specific Ckb knockout animals and for constructive comments on the manuscript together with B. M. Spiegelman  ( Dana-Farber Cancer Institute, Boston ). This work was supported by grants from  Margareta af Uggla 's foundation (M. R. ), the  Swedish Research Council  (M. R. and N. M. ),  ERC-SyG SPHERES  ( 856404  to M. R. ), the  NovoNordisk Foundation  (including the Tripartite Immuno-metabolism Consortium grant number  NNF15CC0018486 , the  MSAM consortium NNF15SA0018346  and the  MeRIAD consortium  grant number  0064142  to M. R. and  NNF20OC0061149  to N. M. ),  Knut and Alice Wallenbergs Foundation  (A. K. , J. R. Z, M. R. and T. M. , including  Wallenberg Clinical Scholar  to M.
Data availability Microarray data generated or retrospectively analysed in this study are publicly available in the NCBI Gene Expression Omnibus repository under the accession numbers GSE25401 (transcriptomics of cohort 2), GSE59034 (transcriptomics of cohort 3), GSE100795 (transcriptomics of different resident cells of human WAT) and GSE192361 (transcriptomics of siCKB-versus siC-transfected cells). Metabolomics data from cohort 1 are provided in ref. 19. RNA sequencing data at multiple time points during adipocyte differentiation are available from these cells within the FANTOM5 project  63  and was used to extract information on genes within the phosphocreatine/creatine pathway presented in Fig. 2b. Source data are provided with this paper. Additional data that support the findings of this study are available from the corresponding authors upon reasonable request.
Applying the sensor temperatures instead of ambient air temperature for the parameterization provides a linear correction function (200-600 W m -2 K -1 s), that depends on the mounting position of the radiometer on HALO. Furthermore, BACARDI measurements from the EUREC 4 A (Elucidating the role of clouds-circulation coupling in climate) field campaign are analyzed to characterize the performance of the radiometers and to evaluate all corrections applied in the data processing. Vertical profiles of irradiance measurements up to 10 km altitude show that the thermal offset correction limits the bias due to temperature changes to values below 10 W m -2. Measurements with BACARDI during horizontal, circular flight patterns in cloud-free conditions demonstrate that the common geometric attitude correction of the solar downward irradiance provides reliable measurements in this typical flight sections of EUREC 4 A, even without active stabilization of the radiometer.
For the installation of BACARDI during EUREC 4 A, two test flights are analyzed, one performed before the campaign in Oberpfaffenhofen, Germany, and one during the campaign in Barbados. In both cases, Θ 0 = +0. 3 • and Φ 0 = +2. 5 • are obtained, so it can be assumed that the offset angles are stable once BACARDI is installed on HALO. To account for the limitations of the attitude correction, the downward solar irradiance is filtered before publishing the data set. Data are assumed to be valid, when the attitude correction factors are less than 25 %. For larger correction factors, roll and pitch angles need to be smaller than 5 •. The excluded data corresponds to turns with large roll angles or conditions with low Sun.
, 2021) , and the frequent dropsonde measurements from HALO  (George, 2021). The absorption by ozone, which becomes relevant for the typical flight altitude of 10 km, is determined by satellite estimates of the atmospheric ozone column. The sea surface albedo is parameterized on the basis of  Cox and Munk (1954) , using the 10 m wind speed obtained from the lower most wind speed value of the HALO dropsondes. A statistically more robust comparison of measured and simulated F ↓ sol is performed merging 12 EUREC 4 flights and filtering the data for cloud-free conditions and reliable attitude corrections. Data are assumed to be valid when the attitude correction factors are less than 25 %. For larger correction factors, roll and pitch angles need to be smaller than 5 •. A one-to-one comparison is shown in Fig.
org); and the 18S rRNA gene sequence database is increasingly large to support operational taxonomic units (OTUs) classification for microbial eukaryotes  (Banos et al. , 2018). For each sample, a 25-µl PCR reaction comprised 1 × EmeraldAmp R GT PCR Master Mix (TaKaRa, Shiga, Japan), 0. 3 µM of each primer, and 75 ng of the metagenome DNA. The 18S rRNA gene PCR also included 10 µM of a mammal-blocking primer (5 -GCCCGTCGCTACTACCGATTGGIIIIITTAGTGAGGCCCT3S pC3-3 )  (Caporaso et al. , 2012). The 16S rRNA gene PCR conditions were 94 • C for 3 min, and 30 cycles of 94 • C for 45 s, 50 • C for 60 s, and 72 • C for 1 min 30 s, followed by 72 • C for 10 min; and the 18S rRNA gene PCR conditions were 94 • C for 3 min, and 30 cycles of 94 • C for 45 s, 65 • C for 15 s, 57 • C for 30 s, 72 • C for 90 s, followed by 72 • C for 10 min.
Bioinformatics and Statistical Analyses Sequences were processed according to Mothur's standard operating procedure (SOP)  (Schloss et al. , 2009). For quality screening, reads containing (i) ambiguous bases; (ii) >1 mismatch base in the reverse primer region, (iii) >10 homopolymer, (iv) sequence length <100 bp, and (v) chimera sequence, were removed. Silva databases (version 1. 32) were used to align the sequences and remove contaminated sequences, such as mitochondria and chloroplast sequences. For taxonomic classification into OTUs, the quality 16S and 18S rDNA sequences were determined using Mothur  (Schloss et al. , 2009)  and Greengenes (version 13. 8); and Good's coverage index was computed to estimate a sequencing coverage (sequencing depth) of each sample. Then, these data samples for 16S and 18S rRNA genes sequences were normalized at 9,045 and 38,429 sequences per sample to prevent a bias from various sequencing depth across samples when comparisons among samples and groups were performed; noted a data sample with fewer sequences than the normalization (samples CAH2-3, CCH1-3, and CRH1-3) was pooled with other independently replicate sample followed by subsampling to normalization or using all those sequences.
Participants were randomly assigned (1:1:1:1) to receive subcutaneous placebo or ianalumab (5 mg, 50 mg, or 300 mg) every 4 weeks for 24 weeks using a secure, online randomisation system. Randomisation was stratified by the ESSDAI score at baseline (≥10 or <10). Study personnel and patients were masked to treatment assignment. The primary outcome was the change in ESSDAI score from baseline to 24 weeks in all randomly assigned patients. Dose-related change in disease activity (ESSDAI) from baseline at week 24 was assessed by multiple comparison procedure with modelling analysis. Safety was measured in all patients who received at least one dose of study drug. This trial is registered with ClinicalTrials. gov, NCT02962895. Findings Between June 27, 2017, and Dec 06, 2018, 293 patients were screened, 190 of whom were randomly assigned (placebo n=49, ianalumab 5 mg n=47, ianalumab 50 mg n=47, ianalumab 300 mg n=47).
Continuous efficacy data were analysed as the change from baseline with a linear mixed effect model for repeated measures, which assumes missing data were missing at random. Binary efficacy data were analysed as the proportion of responders (Clopper-Pearson method), with missing responses treated as non-responders. Efficacy data are shown as least-squares mean with 95% CI. Other data are shown as mean (SD) or mean (SE), or as frequency in percentage. Analyses of baseline features and participation included all randomly assigned patients. Efficacy analyses included all patients assigned to treatment, grouped by the assigned treatment. Safety analyses included all patients who received at least one dose of study drug, grouped by the received treatment. Statistical analyses were done by Novartis using SAS (version 9.
Acknowledgments We thank all patients, investigators, and study personnel for their willingness to participate in the study. The study was funded by  Novartis. Medical writing support was funded by  Novartis. June Mason  drafted the manuscript,  Gurleen Kaur  developed figures, provided editorial support, and managed the submission. Preliminary results of this trial were presented in 2019 at the  American College of Rheumatology , and in 2020 at the  EULAR 2020 Congress  and the  Spanish Society of Rheumatology.
In a second step, this GAM, which represents natural conditions, is applied to predict natural streamflow, i. e. streamflow that would be expected in the absence of the reservoir, for the regulated period. The difference between the observed regulated streamflow signal and the predicted natural baseline should correspond to the reservoir operation signal. We apply this approach to reconstruct the seasonality of reservoir regulation, i. e. information on when water is stored in and released from a reservoir, from a dataset of 74 catchments in the Central Alps with a known reservoir construction date. We group these reconstructed regulation seasonalities using functional clustering to identify groups of catchments with similar reservoir operation strategies. We find that reservoir management varies by catchment elevation, with seasonal redistribution from summer to winter being strongest in high-elevation catchments.
During the unregulated pre-reservoir period, the GAM learns the nonlinear relationship between streamflow time series and corresponding climatological parameters. Then, the estimated transfer function f j calibrated on unregulated periods will be applied via (1) to regulated periods. The difference between the observed regulated streamflow signal and the predicted natural baseline according to our definition corresponds to the reservoir operation signal. We extract this reservoir signal from observed time series of 74 catchments in the Central Alps for which streamflow data are available before and after a known date of reservoir construction (Section 2. 2). From this database of 74 extracted signals, we identify groups of catchments with similar reservoir operation strategies using functional data clustering (Section 2.
Dataset The Central Alps are an interesting region to study different reservoir regulation patterns because this region is characterized by diverse hydro-climatic regimes  (Bard et al. , 2015) , which are often heavily influenced by reservoirs  (Lehner et al. , 2005; Brunner et al. , 2019a). Therefore, we identify a large sample of 74 regulated catchments in the headwater regions of the four major European rivers originating in the Central Alps, namely, the Rhine, Rhône, Danube, and Po for which the date of reservoir construction is known and for which observed daily streamflow data are available for both a period before and a period after reservoir construction (Figure  1 ). The observed streamflow time series were obtained from national agencies in Switzerland (Federal Office for the Environment, FOEN), Austria (Austrian Ministry of Sustainability and Tourism), and eastern France (Banque HYDRO) and regional agencies in southern Germany (regions Bavaria [Bayerisches Landesamt für Umwelt] and Baden-Württemberg [Landesanstalt für Umwelt Baden-Württemberg]).
The streamflow records of the different catchments do not necessarily cover the same time period, however, each catchment has streamflow data for at least 10 years before and after reservoir construction (see Figure  2  for an example time series in the Swiss Alps). Northern Italy was excluded from the analysis because streamflow records provided by the regional agencies did not cover the pre-reservoir construction period. In addition to streamflow, we derive daily meteorological time series (precipitation and temperatures) for each catchment from the gridded E-OBS dataset at 25 km spatial resolution for the period 1950-2020  (Cornes et al. , 2018).
The later signal was never observed but it can be estimated by applying the learning GAM link to post-reservoir meteorological explanatory variables. These covariates include the following three climatological drivers: (1) smoothed daily temperatures, (2) smoothed daily precipitation, and (3) day of the year (seasonality), and interpolated daily glacier mass balance changes (for details on datasets see 2. 1). The last variable takes into account non-stationarities induced by changing glacier melt. Discharges during the natural and regulated period can have different magnitudes as a result of water diversions, e. g. in the case of hydropower production. Therefore, we standardize both the natural and regulated streamflow time series by subtracting the mean. Such standardization makes natural and regulated flow magnitudes comparable.
As positive and skewed random variables, it is unlikely that streamflow time series follow a Gaussian distribution given the four covariates. To handle this issue, we choose a Gamma family within the GAM approach and study the following additive link f 1 (p t ) + f 2 (h t ) + f 3 (d t ) + f 4 (g t ), (2) where p t corresponds to smoothed precipitation, h t to smoothed temperature, d t to the day of the year, and g t to the interpolated glacier mass balance changes (for the implementation, we used the R-package mgcv (Simon  Wood, 2022; Wood, 2017) ). We assess the model's performance by comparing observed with predicted streamflow values (Figure  3 ). ) and used to predict streamflow with precipitation, temperature, day of year, and glacier mass balance changes as predictors. (a) Observed vs.
by averaging over all grid cells within a catchment. Temperature and precipitation time series are smoothed over a moving time window of 5 days to remove noise and NA values are replaced by the mean flow across the whole time period. Furthermore, data on reservoir locations and construction dates are also obtained from national agencies (Switzerland: FOEN, Austria: Austrian Ministry of Sustainability and Tourism, France: Comité Francais des Barrages et Reservoirs (https://www. barrages-cfbr. eu/)) and open source databases (Germany: Speckhann et al. (2021)). To account for changes in glaciermelt contributions over time, we compute annual glacier mass balance changes for each of the selected catchments using simulated mass balance changes over the period 1951-2020 for the glaciers in the Randolph Glacier Inventory (RGI Consortium, 2017; Compagno et al.
inform hydrological model development and calibration. Furthermore, the reconstructed signals could inform the representation of reservoir operation in hydrological models. Improving such representation is crucial to advance the field of change attribution as it will allow for a better separation of climate and regulation signals, which both influence streamflow characteristics. edge the E-OBS dataset from the EU-FP6 project UERRA (https://www. uerra. eu) and the Copernicus Climate Change Service, and the data providers in the ECA&D project (https://www. ecad. eu). Philippe Naveau also acknowledges the support of the French Agence Nationale de la Recherche (ANR) under reference ANR-20-CE40-0025-01 (T-REX project), and the ANR-Melody (ANR-19-CE46-0011). We thank Loris Compagno for providing the simulated glacier mass balance time series.
Methods and findings We conducted a 3-arm cluster-randomized trial (n = 53 clusters) that evaluated the efficacy of (1) prenatal multiple micronutrient supplementation (MMS; n = 18 clusters) and (2) lipid-basedAU : Pleasenotethat}lipid À basedmicronutrientsupplementation}hasbeenchangedto}lipid À basednu nutrient supplementation (LNS; n = 18 clusters) as compared to (3) routine iron-folic acid (IFA) supplementation (n = 17 clusters) among pregnant women in the rural district of Madarounfa, Niger, from March 2015 to August 2019 (ClinicalTrials. gov identifier NCT02145000). Children were followed until 2 years of age, and the Bayley Scales of Infant and Toddler Development III (BSID-III) were administered to children every 3 months from 6 to 24 months of age. Maternal report of WHO gross motor milestone achievement was assessed monthly from 3 to 24 months of age.
Author summary Why was this study done? • It is estimated that over 250 million children under 5 years of age do not reach their developmental potential in low-and middle-income country (LMIC) settings. • Observational studies suggest that poor diet, anemia, and micronutrient deficiencies in pregnancy are associated with poor child development outcomes; however, the effect of macronutrient and micronutrient supplementation in pregnancy on child development outcomes remains inconclusive due to a limited number of randomized trials that have assessed developmental outcomes. What did the researchers do and find? • We conducted a cluster-randomized controlled trial of prenatal multiple micronutrient supplementation (MMS) and lipid-based nutrient supplementation (LNS) as compared Funding: Funding was provided to Epicentre by Medecins Sans Frontières (MSF)-Operational Center Geneva (https://www.
msf. org/) and the Kavli Foundation, Norway (https://www. kavlifoundation. org/). The funding was provided to Epicentre and not individuals. The funders helped to design the study but had no role in data collection or analysis, decision to publish, or preparation of the manuscript. Serum Institute of India Pvt Limited provided vaccine and placebo in kind. Competing interests: I have read the journal's policy and the authors of this manuscript have the following competing interests: RFG is an Academic Editor on PLOS Medicine's editorial board.