dataesr/scientific-paragraphs-categorization huggingface dataset:
Blue points represent the community health site for that fokontany and shaded squares represent residential areas, with larger squares representing areas with more buildings. CHW: community health worker, PHC: primary health center. Data Sources: MMoPH (administrative boundaries), Pivot (health center locations), OpenStreetMap (transportation routes, buildings). All maps have been created from spatial datasets using the R programming language. https://doi. org/10. 1371/journal. pgph. 0001028. g001 the existing site, four had a PHC located within their borders, and the optimal location was in the same residential area as the PHC. For the remaining 30 fokontany, the optimal CHW site placement was in a different geographic location than the current CHW site or PHC (Fig  3 ). The quartile of fokontany with the highest geographic dispersion (above 3.
Discussion Geographic distance to health facilities is one of the main barriers to healthcare access, particularly in rural communities that often live several hours travel time from the nearest clinic. Community health workers (CHWs) have worked for decades within these isolated communities to provide basic health services, especially to children under five years old, and to increase geographic access to primary care. However, even at this finer spatial scale, distance to CHWs may continue to negatively impact healthcare access. A renewed push for community health has translated into updated international guidelines to optimize community health programs in order to maximize their impact, including recommendations to adapt programs to the local geography of CHW catchments. Using a comprehensive transport network dataset, we examined whether geographic barriers persist at the CHW level for child consultation rates in a rural district of Madagascar with a strengthened community health program.
Data Sources: MMoPH (administrative boundaries), Pivot (health center locations), OpenStreetMap (transportation routes, buildings). All maps have been created from spatial datasets using the R programming language. https://doi. org/10. 1371/journal. pgph. 0001028. g003 50% in the most geographically dispersed fokontany. While geographic barriers clearly play a role in limiting healthcare access in this setting, our results suggest that optimizing the location of a single community site will not reduce geographic barriers. Rather, alternative solutions, such as installing additional sites in more dispersed fokontany or the implementation of proactive care programs, may be needed to significantly reduce geographic barriers to community health access. We found that geographic barriers to healthcare access persist in Ifanadiana, even at the level of community healthcare.
The fill color represents the reduction in geographic dispersion and points represent existing site locations (gray) and second site locations (black). CHW: community health worker, PHC: primary health center. Data Sources: MMoPH (administrative boundaries), Pivot (health center locations), OpenStreetMap (transportation routes, buildings). All maps have been created from spatial datasets using the R programming language. https://doi. org/10. 1371/journal. pgph. 0001028. g005 accessibility on primary and secondary healthcare access are well studied (see Guagliardo  [52]  for a review). The findings of this study go one step further and show that geographic isolation remains a barrier for CHW access, where distances to seek care are much smaller. This contributes to a nascent, but growing literature studying the geography of community healthcare  [53, 54] , with the general consensus that geographic barriers also exist at this much finer spatial scale.
Our finding that mixed gender teams have the highest consultation rates suggests that including CHWs of multiple genders at each site is a potential way to increase gender equality without negatively impacting healthcare seeking behaviors, as has been suggested elsewhere  [61]. Additional study, particularly using qualitative methods, is needed to further explore patient perceptions of CHW gender in Ifanadiana. Here, we advance new methods to study the impact of geographic barriers at the community health level, a topic that is currently under-researched due to intrinsic challenges in the spatial resolution of data at these finer scales. Our use of a comprehensive geographic dataset with all the transportation routes and buildings in the district, publicly available on Open-StreetMap, allowed us to examine spatial patterns of healthcare access within CHW catchments with remarkable accuracy.
Ihantamalala et al. [33]  found that these methods more accurately represented residents' travel time than cost-distance maps often used to estimate potential access  [4]. Recent studies of healthcare access  [53, 64]  have made use of fine-scale geographic data from OpenStreetMap or mobile phone data to obtain more accurate estimations of populations' geographical isolation. Several of these data sources are open-source and available online. For example, OpenStreetMap is an open-source dataset of transportation networks that can be viewed via a website (https://www. openstreetmap. org/) or downloaded via the Overpass API (https://overpass-turbo. eu/). In addition, Maina et al. [65]  collated the location of over 98,000 public health facilities from 50 sub-Saharan African countries, which is available through the WHO's Global Malaria Programme (https://www.
who. int/malaria/ areas/surveillance/public-sector-health-facilities-ss-africa/en/). At the primary healthcare level, Weiss et al. [4]  estimated travel times to these health facilities. Given the increasing availability of these data, future studies should leverage this fine-scale data to examine spatial patterns in healthcare inequalities, identifying underserved communities that may be otherwise missed in analyses that aggregate spatial data to coarser scales  [66]. Our study had several limitations. CHW consultation data were reported at the level of the fokontany and individual visits are not georeferenced to each patient's residence. Therefore, our conclusions are limited to inference at the scale of the fokontany, not the individual. Collecting patient-level data would allow for the consideration of more detailed spatial patterns of healthcare utilization within the fokontany, such as identifying a distance threshold for catchment populations where CHW utilization is lower than average.
Acknowledgments We would like to thank  Pivot  staff whose work in Ifanadiana provides the infrastructure that makes this research possible. We are thankful for the contributions of  Vincent Herbreteau ,  Christophe Revilion ,  Jeremy Commin  and the team of local mappers that contributed to the cartography work on OpenStreetMap in Ifanadiana. We would also like to thank the communities of Ifanadiana for their participation in the IHOPE surveys and the community health workers and supervisors for their efforts contributing to data collection in addition to providing primary health care to their communities.
This work was supported by internal funding from  PIVOT  (https://www. pivotworks. org/), which provided salary for TR, MR, FI, LC, GC, KF, FH, LMR, AR, BR, ER, RJLR, and  MHB. It was also supported by a grant from the  Agence Nationale de la Recherche  (Project  ANR-19-CE36-0001-01 ), granted to AG, which supported AG, MR, and  MVE. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
1% TFA was applied for 7 min at a flow rate of 0. 25 mL/min at 29 • C. Detection of hippuric acid was recorded at 228 nm. Peak integration in arbitrary units (AU) was performed using the software LC-Solution ® (Shimadzu, Kyoto, Japan). A calibration curve was generated upstream by injecting increasing amounts of hippuric acid (0-12 nM).
Dynamic Light Scattering and Zeta Potential Measurements The size of the liposomes and their zeta potential were measured through dynamic light scattering (DLS) and electrophoretic light scattering (ELS) using a Zetasizer Nano ZS from Malvern Instruments ® (Malvern, UK). The protocol was adapted from Hasan et al. [39]. Empty liposome and SHH2 and SHH3 liposome samples were diluted to one-five-hundredth (1:500) in ultrapure water and then were spotted with a syringe into a zeta cell. An algorithm built into the Malvern ® Zetasizer software (Malvern Instruments, Malvern, UK) analyzed the measured intensity autocorrelation functions. All measurements were made at 25 • C.
Isolation and Culture of Human MSCs Umbilical cords were collected at the Maternity Hospital of Nancy after obtention of a consent form signed by the pregnant mothers in compliance with the French national legislation regarding human sample collection, manipulation and personal data protection. This collection was approved by the Nancy Hospital Ethics Committee and French Ministry of Higher Education, Research and Innovation (registration number no. DC2014-2114). Human umbilical cords (~20 cm) were obtained from full-term births (n = 3). Human Wharton's jelly MSCs (MSCs) were isolated and expanded in culture according to standard protocols. Briefly, umbilical cord vessels were removed from cord segments, and exposed mesenchymal tissue was cut into small pieces (1-2 mm 3 ) that were treated for 18 h with collagenase type 2 (1 IU/mL) (ThermoFisher, Walthma, USA) in alpha Minimal Eagle Medium (α-MEM) (Lonza, Basel, Switzerland) at 37 • C with 5% CO 2 and 95% humidity.
The membrane was incubated overnight (4 °C) with the primary antibody, was rinsed, and then incubated for 2 h with the appropriate horseradish peroxidase-conjugated secondary antibody (Sigma-Aldrich, Saint Quentin Fallavier, France). Peroxidase activity was revealed by using enhanced chemiluminescence (ECL) reagents (Luminata Crescendo, Millipore), Molsheim, France. The intensity of peroxidase activity was quantified using Image-J software (NIH, Bethesda, USA). Beta-tubulin (β-tub) and beta-actin were used as loading controls for GFAP and dPP2Ac quantification, respectively, whereas the total protein was used for phosphorylated protein isoforms. The following antibodies were used: mouse anti-GFAP (1/1000, Sigma Aldrich Saint Quentin Fallavier, France), mouse anti-phospho Tau (phospho-threonine 231, clone AT180, 1/1000, Thermofisher, Illkirch Graffenstaden, France), mouse anti-phospho Tau CP13 (phospho-serine 202, 1/1000 gift from Dr.
Oxylipins Quantification Non-enzymatic oxygenated polyunsaturated fatty acid metabolites (NEO-PUFAs) were extracted from prefrontal cortex tissue or microemulsions as previously published  [35]. The qualitative and quantitative profile of these non-enzymatic lipid mediators was determined in the mouse brain by micro-LC-MS/MS as previously described  [36]. Their concentration was established by calibration curves calculated from the area ratio of analytes and the internal standards. Data processing was performed using a MultiQuant 3. 0 software (Sciex Applied Biosystems, Framingham, MA, USA).
A metaanalysis concluded that the ACE2 variant rs190509934:C (a rare variant) characterized by a lower ACE2 expression in individuals carrying the C allele, reduces the risk of SARS-CoV-2 infection  (Horowitz et al. , 2021). Analysis of inter-individual ACE2 polymorphism, based on broad genomic databases reveal a link with the susceptibility to SARS-CoV-2 and the severity of COVID-19  (Brest et al. , 2020;   Cao et al. , 2020). The pioneering work by Cao and colleagues identified 15 unique expression quantitative trait loci variants (14 SNPs and 1 InDel) with a higher frequency of minor alleles in the Asian population than in the European population. For example, the rs143695310 variant among East Asian populations was found to be associated with elevated expression of ACE2. Moreover, it was reported that Asian men have a higher ACE2 mRNA expression in their lungs than women, and that Asian people express higher amount of ACE2 than Caucasian and African American populations according to single-cell RNA-seq analysis  (Zhao et al.
4 mg/kg i. v. ), was found to be well-tolerated and to have potential haemodynamic benefits in pulmonary arterial hypertension  (Hemnes et al. , 2018). A hrsACE2 clinical trial is ongoing (NCT00886353) for the treatment of cardiovascular diseases  (Ghatage et al. , 2021). It has been reported that ACE2 S 680 D gain-function knock-in mice are protected against hypoxia-induced pulmonary hypertension  (Zhang et al. , 2018). This has also opened the way to modified ACE2 for gene transfer  (Guignabert et al. , 2018). It was recently demonstrated that recombinant ACE2 is effective for treating SARS-CoV-2 RBD protein-aggravated LPS-induced acute lung injury in a mouse experimental model and that the protection occurs by acting on the ACE2-AngII-AT1R-NOX1/2 axis that is otherwise overactivated by the SARS-CoV-2 infection  (Zhang et al.
Funding This work was supported by the  French Government  under the " Investissements d'avenir" (Investments for the Future) program  managed by the  Agence Nationale de la Recherche (French ANR: National Agency for Research ; reference: Méditerranée Infection  10-IAHU-03  to  Professor Didier Raoult ), and annual funds from  Aix-Marseille university and IRD  to the MEPHI research unit (Director:  Professor Jean-Christophe Lagier ). Other funding sources were limited to the salaries of the authors ( Center National de la Recherche Scientifique for CAD, Assistance Publique Hôpitaux de Marseille for LCJ ), with no other role or involvement.
3 3 FIGURE 2 3-D model of ACE2 protein structure according to the published data PDB: 6M1D by Yan and colleagues in 2020. ACE2 is composed of 805 amino acids and characterized by an N-terminal signal peptide of 17 amino acid residues, a peptidase domain (PD; residues 19-615) with its HEXXH zinc binding metalloprotease motif, a C-terminal collectrin-like domain (CLD; residues 616-741) that includes a ferredoxin-like fold "neck" domain (616-626), that end with a hydrophobic transmembrane hydrophobic helix region (741-763) followed by an intracellular segment of 43 amino acid residues. No information is available regarding the 3-D structure of the ACE2 cytoplasmic tail (763-805), which was drawn to appear on the diagram. Some of the amino acids which are considered important for SARS-CoV-2 interaction are located in the 3-D model (amino acids in red).
The role of TRPM7 was studied by RNA silencing or by pharmacological inhibition. TRPM7 expression was found to be correlated with the activation status of PS-1 cells. TRPM7 expression was able to regulate proliferation through modulation of cell cycle regulators and most importantly p53, via the PI3K/Akt pathway, in a magnesium-dependent manner. Finally, the analysis of TCGA database showed the overexpression of TRPM7 in cancer-associated fibroblasts. Taken together, we provide strong evidences that TRPM7 can be considered as a marker of activated PSCs.
Cell Cycle Analysis Cell cycle analysis was conducted using DNA cellular content quantification by flow cytometry. 1 × 10 6 PS-1 cells were fixed with ethanol and sorted out by a cytometer (Accuri C6, BD Biosciences, Rungis, France). The percentage of cells in each phase of the cell cycle was obtained by calculating the area under the curve using the Cyflogic version 1. 2. 1 software (CyFlo Ltd. , Turku, Finland).
Analysis of CAF Datasets Analyses of relative expression of TRPM7, TRPM6 and POSTN were conducted using GEPIA tool (http://gepia. cancer-pku. cn/, accessed the 2 March 2022) that integrates a normal pancreas from GTEx (n = 171) and pancreatic cancer PAAD dataset from TCGA (n = 179). Expression of TRPM7, TRPM6 and POSTN in different cell types were analyzed using GEPIA2021 and EPIC deconvolution tools (http://gepia2021. cancer-pku. cn/, accessed the 2 March 2022)  [35, 36]. Quantitative comparisons of the cell proportions or expression in different cell types were analyzed using built-in t test (GEPIA) or ANOVA test (GEPIA2021). The GSE28735 dataset contains 45 normal pancreases (adjacent non tumoral, ANT) and 45 tumor (T) tissues from PDAC cases. mRNA levels were analyzed using the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database (http://www.
Statistical Analysis Data are presented as Mean ± S. E. M. and the number of separate experiments or the number of studied cells is represented by n. Experiments were repeated in at least three different cell passages. Data analysis and figure conception were made by using Microcal™ Origin ® (OriginLab Corporation, Northampton, MA, USA), Clampfit (Molecular Devices, Inc. , San José, CA, USA) and Inkscape software. Statistical analyses were made using Student's -test or Mann-Whitney rank sum test depending on sample normality determined by paired Wilcoxon signed rank test. A two-way analysis of variance was used followed by the Holm-Sidak method to compare two parameters or more. All the statistical analyses were conducted using Sigma-Stat 3. 0 (Inpixon, Palo Alto, CA, USA).
TRPM7 Expression Is Increased in Cancer-Associated Fibroblasts (CAFs) We investigated the expression of TRPM7, TRPM6 and periostin (POSTN), a marker of activated CSPs  [13, 44] , in pancreatic cancer samples using GEPIA tool. We observed that POSTN mRNA relative level was increased in tumor samples when compared with a normal pancreas (p < 0. 05, Figure  7A ). We also observed a trend toward an increase in TRPM7 that was not statistically significant. TRPM6 expression was not altered in PDAC. Spearman correlation analysis showed that TRPM7 and POSTN were positively correlated in tumor samples (r = 0. 24; p = 0. 0013, Figure  7B ). We also observed similar findings in an independent PDAC dataset (GSE28735), in which POSTN mRNA level was increased in tumor sample compared to adjacent non tumoral tissue (not shown).
Funding: This research was funded by The  French National Research Program for Environmental and Occupational Health of Anses  with financial support from  ITMO Cancer of Aviesan  on funds administered by  Inserm  ( 2020/01/195 );  Le Ministère de l'Enseignement Supérieur, de la Recherche et de l'Innovation; La Ligue Contre le Cancer ; and  FEDER. Julie Auwercx  and  Thibaut Lefebvre  are recipients of a PhD fellowship from  Université de Picardie Jules Verne. Silviya Radoslavova  was a recipient a PhD fellowship from  Région Hauts-de-France  ( 50% ) and from  Université de Picardie Jules Verne  ( 50% ). Alison Vanlaeys  was a recipient of a PhD fellowship from  Région Hauts-de-France. Institutional Review Board Statement: Not applicable. Informed Consent Statement: Not applicable.
Data Availability Statement: TCGA, GTEX and GSE data are available and are based upon public data extracted from TCGA Research Network. Available online: http://cancergenome. nih. gov/ (accessed the 2 March 2022), Genome Tissue Expression (GTEX) project. Available online: http:// www. GTEXportal. org/ (accessed the 2 March 2022), and Gene Expression Omnibus (GEO) database. Available online: http://www. ncbi. nml. nih. gov/geo/ (accessed the 2 March 2022).
Behavioral Evaluation For behavioral studies, QuilA-EAE mice and CTL were tested at disease onset (D17) and during the chronic phase of the disease (D30). In the 14 behavioral procedures, the number of tests for each animal was limited to 4-7 according to the severity of the procedures. Also, to take into consideration the variability of EAE induction (28-30), each behavioral evaluation was always performed in two different cohorts of EAE. Hence, seven cohorts (groups of animals immunized at the same time and evaluated with the same tests) were used (each cohort included 12 CTL and 12 EAE mice except for Cohort 4, in which only 10 animals were included in each group). This procedure is summarized in Table  1. Body weight, RR-EAE score, and von Frey evaluation were determined in all seven cohorts, but for simplicity, we show data from only two cohorts closely representative of the others.
Motor Evaluation Spontaneous locomotion activity was evaluated using the open field test. Mice were placed in the center of a white polyvinyl chloride open field apparatus (50 cm long × 50 cm wide × 45 cm high) and allowed to move freely. Virtual areas, a central square (30 cm long), and a peripheral zone were determined with videotracking software (Viewpoint, Lyon, France). For 15 min, total distance traveled and time spent in the central area were recorded with the videotracking system. Motor coordination was determined using a standard mouse rotarod (TSE, Bad Homburg Germany). We used the standard operating procedures described by the EUMORPHIA group program  (31). Briefly, after a training phase to identify mice able to stay on the rod at 4 rpm for 60 s, mice underwent the test phase. This consisted of four trials with a 15 min intertrial interval.
Mice were placed on a 50 cm long × 50 cm wide wire grid floor (wire diameter 1. 5 mm, grid mesh 1 × 1 cm) 30 cm above an infrared floor. Locomotor activity was recorded by the first camera. The experiment was stopped when the mice had traveled a distance of 300 cm. The number of times that mice slipped on the wire mesh floor was counted by an experimenter from recordings made by the second camera. Muscular strength was measured for forelimbs or both hind and forelimbs with a grip strength meter (Bioseb, Chaville, France). We used the standard operating procedure described by the EUMORPHIA group program  (31). Mice were lifted and held by their tails so that their limbs could grasp a wire grid. The mice were then gently pulled backward by their tails with their posture parallel to the surface of the bench until they released the grid.
Working memory was evaluated using the Y-maze test according to the standard operating procedure described by the EUMORPHIA group program  (31). Spontaneous alternation behavior and exploratory activity were recorded. The apparatus was made of three equal arms (40 cm long × 10 cm wide × 16 cm high) made of black polyvinyl chloride radiating at 120 • from each other. Mice were placed at the end of one arm, facing the end wall of the arm, and allowed to move freely through the maze for a 10 min session. Latency to leave the first arm and total number and sequence of entries into each arm were scored for each mouse. An arm entry was counted when the mouse had all four paws inside the arm. If mice completed fewer than eight arm entries within 10 min, they were excluded from further analysis. Spontaneous alternation was defined as entries into all three arms on three consecutive choices.
Statistical Analysis All statistical analyses were carried out using GraphPad Prism 6 software (GraphPad Inc. , San Diego, USA). Before each analysis, equality of variance and normal distribution were evaluated, and parametric or non-parametric analyses were performed according to the results observed. For body weight and EAE score longitudinal monitoring, data were analyzed using two-way ANOVA with group (CTL vs. QuilA-EAE) and day post-induction (DPI) (D0-D40) as main factors with DPI being defined as a repetitive measure. Post-hoc comparisons were made with a Sidak test for multiple comparisons between groups for each DPI. For motor and sensitivity behavioral tests, data were analyzed using two-way ANOVA with group (CTL vs. QuilA-EAE) and DPI (D17 vs. D30) as main factors, with DPI being defined as a repetitive measure.
Extended Characterization of Sensitive Behaviors In most studies evaluating somatic sensitive behaviors in EAE mice (whatever the variant used), mechanical allodynia was always assessed, whereas heat hyperalgesia was less often studied. Studies assessing the latter parameter showed divergent results for the acute EAE phase of the disease, with some demonstrating heat hypoalgesia (29,  44, 45)  and others heat hyperalgesia  (46) (47) (48). Data related to the chronic phase of the disease were more closely consistent with all the studies showing heat hyperalgesia in EAE mice. The results obtained in our study showed for the first time that QuilA-EAE mice developed heat hyperalgesia during the chronic phase of the disease. Concerning cold allodynia, in our study, it occurred in the early phase of the disease.