dataesr/scientific-paragraphs-categorization huggingface dataset:
The study included 541 patients with GPP and reported a GPP case estimate of 7. 46 patients per million, with 2. 87 patients per million presenting with acute GPP  [33]. A recent analysis of the Japanese national database of health insurance claims suggests this figure could be higher. The database covers approximately 90% of Japanese residents and recorded 4636 codings of pustular psoriasis, which would indicate over 30 cases per million people  [35]. A review of a national insurance claims database in the Republic of Korea conducted between 2011 and 2015 estimated a prevalence of 1. 2 cases per 10,000 persons. In patients with psoriasis (range 219,429-233,909 across study years), 2. 0-2. 7% were categorized as having GPP. The authors equated prevalence in the database to national prevalence because of compulsory enrollment in the insurance database for all residents of the Republic of Korea  [34].
A US insurance claims review was conducted between 2016 and 2019 using the IQVIA PharMetrics® Plus database. Patients were required to be continuously enrolled in the medical and pharmacy benefits for 6 months before and at least 2 months after the index date (date of diagnosis). This database analysis identified 990 patients with GPP over the study period  [36]. It should be noted that the large variations reported across the published studies do not necessarily transcribe an epidemiological reality, and that available data should perhaps be considered as indicative of prevalence only. The variability is likely driven by challenges related to the study of a rare and relatively unknown disease, as well as differences in the distribution of genetic variants and cofactors in separate populations, and reported estimates represent a range that must be interpreted while considering methodological differences.
As described, the considerable interstudy variation in methodologies and populations means that physicians have an incomplete view of the epidemiology of GPP. It is likely that determining national population-level prevalence estimates will remain challenging for some time. However, database assessments can provide specific insights into certain aspects of GPP, including regional variations, and there is value to the pustular psoriasis community in continuing to generate insights into aspects of GPP epidemiology. While database studies cannot act as proxies for accurate estimates of disease prevalence in the wider population, they can contribute to disease understanding. In Japan, data were collected for 718 patients with GPP and 27,773 patients with plaque psoriasis between January 2015 and December 2019 using the Japanese Medical Data Vision database.
As with many rare diseases, forming a clear picture of the clinical characteristics and disease prevalence is challenging. Key barriers include limited disease awareness and the different approaches to assessing prevalence, which are often performed using database studies or small hospital audits. In addition, the data gained from these initiatives are not commonly extrapolated to a national population-level prevalence. Despite the inherent limitations of medical database analyses, these studies can contribute up-to-date insights into various aspects of GPP, including disease characteristics and prevalence in defined cohorts. medical writing, editorial and/or formatting support, which was contracted and funded by Boehringer Ingelheim. Boehringer Ingelheim was given the opportunity to review the manuscript for medical and scientific accuracy as well as intellectual property considerations.
Kinetic analysis Kinetic rate constants (k a and k d ) of monoclonal AIGAs to IFN-γ were measured by BLI (FortéBio Octet RED96). The running buffer used contained 0. 1% BSA, 0. 1% Tween-20, 250 mM NaCl, 2. 7 mM KCl, 10 mM Na 2 HPO 4 , and 1. 8 mM KH 2 PO 4 in sterile water. Monoclonal AIGAs (1 μg/ml) were loaded onto antihuman IgG Fc capture biosensors (FortéBio) to reach a wavelength shift range of 0. 5∼2. 0 nm and were then exposed to various concentrations of IFN-γ in running buffer for the association step, before dissociation in running buffer. Rate constants were calculated by fitting a curve (1:1 Langmuir model) of binding response, with the 5-min association and 15-min dissociation interaction times, with Octet Data Analysis software v9. 0. In-tandem cross-competition assay Biotinylated IFN-γ (biotin-IFN-γ, 2 μg/ml) was loaded onto the streptavidin biosensor (FortéBio) to trigger a saturated response.
HLA expression of THP-1 cells Monoclonal AIGAs or control IgG (3 μg/ml) were incubated with or without IFN-γ (20 ng/ml) for 30 min. We then cultured 1 × 10 5 THP-1 cells (BCRC-60430) in 96-well culture plates, in 200 µl of RPMI-1640 medium supplemented with 10% FBS, and 1% penicillin/streptomycin for 24 h, with or without stimulation. The cells were washed twice with 1% FBS in PBS. The stimulated and unstimulated THP-1 cells were stained with anti-human HLA-DR-PE antibody (BD Pharmigen) and incubated on ice in the dark for 1 h. The stained cells were washed with 2 ml of 1% FBS in PBS and resuspended in 500 μl of 1% FBS in PBS. The fluorescence intensity of the stained cells was acquired with a FACSVerse flow cytometer (BD Biosciences) and analyzed with FlowJo software v10 (BD Biosciences).
Cell-based antibody-binding assay We incubated 1 × 10 5 THP-1 cells with 2. 5 μg Fc Block (BD Biosciences) in 100 μl RPMI-1640 supplemented with 10% FBS and 1% penicillin/streptomycin for 15 min. We incubated monoclonal AIGAs or control IgG (40 nM) with or without IFN-γ (20 nM) at 25°C for 15 min. The mixtures were then added to the THP-1 cells, which were incubated at 37°C for 15 min. The cells were washed with 2 ml of 1% FBS in PBS, stained by incubation with anti-human IgG-PE (BD Pharmigen) for 30 min at 4°C, and washed with 2 ml of 1% FBS in PBS. The cells were resuspended in 200 μl of 1% FBS in PBS, and the data were collected with a FACSVerse flow cytometer and analyzed with FlowJo software v10.
Acknowledgments We thank the patients' families for their trust and cooperation. We thank  S. -C. Jao  and  W. -C. Huang  ( Academia Sinica ),  Z. -Q. Gu  and  Y. -S. Chu  ( National Yang Ming Chiao Tung University ), and  C. -L. Hsiao  and  Y. -T. Kuo  ( Elixiron Immunotherapeutics ) for providing advice concerning experiments. We thank the  Flow Cytometric Analyzing and Sorting Core Facility at National Taiwan University Hospital. We thank  Mark R. Walter  ( University of Alabama at Birmingham, Birmingham, AL ) for providing a plasmid for producing IFN-γ. The study was funded by  Chang Gung Memorial Hospital  ( CMRPD1K0681-2  and  BMRPB98 ), the  Taiwan Ministry of Science and Technology  ( 105-2628-B-182-002-MY3 ) and the  Deutsche Forschungsgemeinschaft  ( PI 405/14-1  and  INST   190/ 146-3 ).
Background Inebilizumab is an anti-CD19 antibody approved for the treatment of neuromyelitis optica spectrum disorder (NMOSD) in adults with aquaporin-4 autoantibodies. The relationship between B-cell, plasma-cell (PC), and immunoglobulin depletion with longitudinal reductions in NMOSD activity after inebilizumab treatment was characterised post hoc in an exploratory analysis from the N-MOmentum study (NCT02200770). Methods Peripheral blood CD20+ B cells, PC gene signature, and immunoglobulin levels were assessed throughout N-MOmentum (follow-up ≥2. 5 years); correlations with clinical metrics and magnetic resonance imaging (MRI) lesion activity were assessed. Findings Inebilizumab induced durable B-cell and PC depletion within 1 week versus placebo. Although no association was observed between B-cell counts at time of attack and NMOSD activity, depth of B-cell depletion after the first dosing period correlated with clinical outcomes.
Study design and participants The N-MOmentum study was an international, multicentre, double-blind, randomised, placebo-controlled, phase 2/3 trial with an open-label extension (ClinicalTrials. gov identifier: NCT02200770). 19 Adults (aged ≥18 years) with NMOSD were eligible for inclusion if they had an Expanded Disability Status Scale (EDSS) score of 8. 0 or less, and a history of at least one attack requiring rescue therapy in the previous year, or of at least two attacks requiring rescue therapy in the previous 2 years. AQP4-IgG serostatus was determined centrally using the Euroimmune cell-based assay as described. 19 Patients who were AQP4-IgG seropositive and those who were AQP4-IgG seronegative according to the 2006 neuromyelitis optica diagnostic criteria were eligible. 20 Further study details can be found in the Supplementary Methods; full study details were published previously.
Outcomes Data were captured in a validated electronic system, and processed (including cleaning, reconciling, and quality control processing) and stored in a controlled access environment. Clinical data were analysed according to the approved statistical plan, available at https:// clinicaltrials. gov/ProvidedDocs/70/NCT02200770/SAP_ 001. pdf. The primary efficacy endpoint was the time to onset of an adjudicated NMOSD attack during the RCP. An attack was defined as the presence of one or more new symptoms, or worsening of one or more existing symptoms related to NMOSD, that met at least one of the 18 protocol-defined neurological examination criteria for an attack. These criteria comprised myelitis, optic neuritis, and the brain/brainstem domain symptoms. 21 The study investigator evaluated all potential attacks within 72 hours.
Evaluation of B-cell pharmacodynamics During the RCP, blood samples were collected during study visits at baseline and weeks 1, 2, 4, 8, 12, 16, 22, and 28 to assess B-cell counts; at baseline and weeks 2, 4, 8, 12, 16, and 28 to evaluate plasma-cell-specific gene expression; and at weeks 12 and 28 to evaluate Ig levels. During the OLP, blood samples were collected every 13 weeks for assessment of B-cell counts, plasma-cellspecific gene expression, and Ig levels. Data on T-cell subsets were not collected. Blood samples to assess Bcell counts and plasma-cell-specific gene expression were also collected during any assessment visit for new or worsening NMOSD symptoms during the RCP and OLP. Samples for attack assessments were taken before initiation of treatment for NMOSD attacks. B-cell levels in peripheral blood were measured by flow cytometry using fluorescence-activated cell sorting (FACS), conducted at a central laboratory in the 72 hours after sample collection.
Statistics The analyses presented in this article are exploratory and p values are provided for hypothesis generation only. Descriptive statistics were calculated for changes in B-cell subset counts over time. The statistical analyses were conducted in R 4. 1. 3. Differences in B-cell counts, total Ig levels, and plasma-cell signatures were assessed using the Mann-Whitney U test. Relationships between depletion groups and treatment outcomes for summarisation were estimated by negative binomial regression (details are included in the Supplement) or Poisson regression when appropriate, and included an offset term to account for the varying intervals of time for which participants were included in the RCP. Glm. nb from MASS package in R. 4. 1. 3 was used to fit the model. Differences in gene expression, including FCGR3A F/F genotype between depletion groups, were assessed using Fisher's exact test.
These values are well above the mutation significance cutoff (MSC), with a 99% confidence interval of 25. 1 for PRKCD. The variants are private and have never been reported in public databases such as gnomADv2. 1. 1 and v3. 1. 2, ExAC, BRAVO/Topmed, or ATAV or in patients with AR PKCδ deficiency (Fig. 1e ). Target panel sequencing also identified a private intronic variant, c. 376 + 6A > G, in the same allele as p. D126Y. This variant had a low CADD score (15. 28), was not predicted to abolish splicing, and is, therefore, probably not disease-causing. However, biallelic variants of PRKCD have been reported to underlie susceptibility to infections similar to those observed in patients with CGD  [5]. Taken together, these results suggest that the patient is suffering from AR PRKδ deficiency.
The Patient's Variants Impair Splicing and Are Loss-of-Expression Both potentially deleterious variants are located in the N-terminal part of the PKCδ protein, between the C2-like and C1 domains (Fig. 1f ). The p. D126Y variant is located at the end of exon 5 and is predicted, by GeneSplicer, NNSPLICE, and MaxEntScan, to disrupt the 5′ splice donor site. We studied the potential impact of the variants on mRNA splicing by performing TOPO-TA cloning and sequencing on cDNA from MDMs from the patient and a healthy control. Three different transcripts were detected for the patient, all of which were absent from the healthy control: 30% of the transcripts corresponded to the nonsense variant p. C95*, whereas 60% of transcripts displayed a skipping of exon 5 (c. 316_376del) due to aberrant splicing and were predicted to encode a truncated protein (p.
CD49f, EGFR, PD-L1, HLA-G, TGFB and IL-10 expression profiles were analyzed using an Astrios cell-sorter (Beckman Coulter, Villepinte, France) or Attune NxT (Thermo Fisher, Les Ulis, France) or MACSquant (Miltenyi, Paris, France) analyzer. Data were analyzed using FlowJo software (BD Biosciences, Le Pont de Claix, France).
Keratinocyte Precursors Overexpress the Immune Checkpoints HLA-G and PD-L1 We then investigated the mechanisms by which keratinocyte precursors inhibit CD4 + T-cell proliferation. By high content single-cell image analysis, we observed that 7-days amplified keratinocytes expressed HLA-G and PD-L1 immune checkpoints (Figures  2A, C ). When HLA-G and PD-L1 were analyzed according to CD49f expression, we found that amplified keratinocytes that had the highest levels of CD49f overexpressed both HLA-G and PD-L1 (mean ± SEM, p<0. 0001, n=3) (Figures  2B, D ). These data were confirmed by flow cytometry (Figures  2E, G ). Furthermore, keratinocytes were analyzed for expression of the immune suppressive cytokines IL-10 and TGFB. No IL-10 expression was observed. TGFB expression was found in all keratinocyte subpopulations, but no difference in expression levels was observed.
Exploration was defined as the orientation of snout of the animals toward the object, sniffing or touching  (Antunes and Biala, 2012; Ettcheto et al. , 2016). In those cases, when the total time exploring both objects was less than 5 min, the mouse was excluded. Data were measured by discrimination index (DI), which indicates the difference in exploration time in seconds between familiar and novel object, using the followingt equation:.
Finally, results were obtained through chemiluminescence detection using the Pierce ® ECL Western Blotting Substrate ( # 32106, Thermo Scientific, United States), a Bio-Rad Universal Hood II Molecular Imager, and the Image Lab v5. 2. 1 software (Bio-Rad laboratories). Measurements were expressed in arbitrary units and all results were normalized with the corresponding loading control (Glyceraldehyde-3phosphate dehydrogenase; GAPDH).
Statistical Analysis All results are presented as MEAN ± SD. Groups were compared against each other using two-way ANOVA. When variables independently were significant, we denoted # p < 0. 05, ## p < 0. 01, ### p < 0. 001, and #### p < 0. 0001 for the diet factor and $ p < 0. 05, $$ p < 0. 01, $$$ p < 0. 001, and $$$$ p < 0. 0001 for the genotype factor. When the interaction between two analyzed factors in ANOVA was significant, Tukey's post hoc test was performed for comparison among groups (pp < 0. 05. ppp < 0. 01, pppp < 0. 001, and ppppp < 0. 0001). All analyses and GAPDH representations were obtained using Graph Pad Prism software for Mac version 6. 01; Graph Pad Software, Inc.
In our study, the dendritic spine detection and subsequent quantification were performed where the obtained results correlated with those of the behavioral test. Specifically, our data demonstrated a significant effect of diet and genotype (p < 0. 01 and p < 0. 0001, respectively) with the interaction between both variables (p < 0. 01). Following, Tukey´s post hoc was performed and our results showed a significant decrease of 30% in hippocampal spine number in WT mice after being fed with HFD (p < 0. 0001). By contrast, animals with PPARβ/ δ deficiency also exhibited a significant reduction compared to WT, but HFD feeding did not induce a synergic effect (WT CT vs. PPAR β/δ -/-CT 22%, p < 0. 001; WT CT vs. PPAR β/δ -/-HFD 30%; p < 0. 0001) (Figures  3A, B ). Moreover, as can be observed in Figure  3A , the reduction in spine number was accompanied by shorter and smaller dendritic spines (qualitative evaluation).
Simultaneously, we assessed by GC-MS the concentration of 87 metabolites detectable by our platform of metabolomics across most of our plasma samples with satisfactory quality controls. The whole work was made as an ancillary study of the TopNivo trial (NCT03226756). We found that galectin and metabolite distribution was dependent on disease categories defined according to the primary tumor site and the HPV status. High concentrations of gal-9 and kynurenine (Kyn) were recorded in plasma samples related to hypopharyngeal carcinomas. In contrast, high concentrations of fatty acids were recorded in samples related to carcinomas of the oral cavity. This is a reminder that inside the vast group of HPV-negative HNSCCs, there are distinct types of tumor macroenvironments. High plasma levels of gal-9 appear to be characteristic of advanced hypopharyngeal carcinomas.
Patients and healthy controls TopNivo is a safety multicentric study of Nivolumab in patients with recurrent and/or metastatic platinum-refractory HNSCCs. Nivolumab injections were given every 2 weeks, up to 12 cycles (1 cycle = 28 days). A total of 351 patients have been enrolled in TopNivo (NCT03226756). In the last stage of the clinical trial recruitment, EDTA blood samples were obtained from 83 consecutive patients, at baseline, prior to the onset of Nivolumab. Plasma samples from anonymous healthy donors were used as controls. They were purchased from Zenbio (NC, USA )(ages comprised between 19 and 65). The HPV status of oropharyngeal carcinomas was determined according to p16 expression detected by immunohistochemistry.
Samples were then transferred in vials and directly injected into the GC-MS system. Sample analysis. GC-MS/MS was performed with a 7890A gas chromatograph (Agilent Technologies, Waldbronn, Germany) coupled with a triple quadrupole 7000C (Agilent) equipped with a high sensitivity electronic impact source (EI) operating in positive mode. This method was inspired from developments reported in  (Tsugawa, Tsujimoto et al. 2014 ). The scan mode was the MRM for biological samples. Peak detection and integration of the analytes were performed using the Agilent Mass Hunter quantitative software (B. 07. 01)  (Viltard, Durand et al. 2019). All targeted treated data were merged and cleaned with a dedicated R package (version 4. 0)(@Github/Kroemerlab/GRMeta).
The same plasma samples used for galectin assays were subjected to this procedure. After preliminary runs, 87 out of 130 metabolites were selected for subsequent analyses, based on their detection in the vast majority of plasma samples with satisfactory quality control (list in Supplementary Table  5 ). A correlation map including the 87 metabolites and the 5 proteins (gal-1, -3, -9, CRP and CXCL9) was built using the MetaboAnalyst software (Figure  4 ). Five main clusters were delimited on this map according to the boundaries of color contrasts and, to a lesser extent, the arborescence of the dendrogram. They were designated A, B, C, D, E (details on their content are provided in the legend of Figure  4 ). Briefly, the two biggest clusters, A and B, mainly consisted of free proteinogenic amino-acids.
proteins in connection with tumor/disease categories. This heatmap was generated using the "heatmap" function of the MetaboAnalyst software. The color scale is a function of biomolecule concentrations (m/z peak areas for metabolites and pg/ml or ng/ml for proteins, all normalized to the corresponding maximal concentration recorded in the 83 sample series). For better view and reading, the heatmap was split in two parts. "Oro" stands for oropharyngeal carcinomas. Overall, according to Anova statistical analysis, none of the metabolites on this map showed a significant difference of distribution between the 5 disease categories. However, it is noteworthy that a large number of metabolites (38/87) were at higher relative concentrations in plasma samples related to hypopharyngeal carcinomas. Most components of cluster E identified in Figure  4 , especially one protein (gal-9; bold blue arrow) and several metabolites (kynurenine, arabitol, threonic acid; thin blue arrows, part 2)) were abundant in samples from hypopharyngeal carcinomas.
Single-cell RNA-sequencing Single-cell RNA-sequencing (scRNAseq) was performed on peripheral blood mononuclear cells (PBMCs) from the index patient (R291X) and 4 controls using the 103 Genomics Chromium Controller with the Chromium Single Cell 3  9  Reagent Kit (v3 Chemistry) following the manufacturer's instructions. After quality control, libraries were sequenced on the Illumina HiSeq 4000 platform in 2 3 75 bp paired-end mode. Supplemental Table  1  includes an overview of sequenced samples. More details are provided in the data supplement.
Clinical phenotype and identification of IKZF2 variant We studied a 16-year-old girl born to healthy nonconsanguineous parents (Figure  1A ) who at 10 years of age developed skin lesions, photosensitivity, polyarthritis, and mild ITP with positive antiplatelet, anti-DNA, and antinuclear autoantibodies, prompting a diagnosis of SLE. The patient was in clinical remission upon treatment with hydroxychloroquine and nonsteroidal anti-inflammatory drugs (supplemental Table  1 ). To investigate a potential monogenic etiology of the disease, we performed whole-exome sequencing and identified a paternally inherited heterozygous variant in IKZF2 located after the DNAbinding domain that results in a premature stop codon in exon 8 (c. 871C. T, p. Arg291Ter; Figure  1B ). Stringent filtering revealed that the variant was ultrarare (Genome Aggregation Database [gno-mAD] minor allele frequency, 4 3 10 26 ) and predicted to be deleterious (Combined Annotation Dependent Depletion score of 38; supplemental Table  2 ).
Identification of additional patients with Helios mutation Stimulated by the effect of the Helios R291X variant, we revisited our database of patients with IEIs. We identified 5 additional patients from 4 nonconsanguineous families carrying 4 different rare (gnomAD allele frequency ,0. 001) heterozygous missense variants in IKZF2, which were predicted to be damaging (c. 1076A. G, p. Y359C; c. 1038G. A, p. V347M; c. 316C. T, p. R106W; and c. 659A. G, p. N220S, respectively; supplemental Figures  1B-C ). Intriguingly, these patients presented with profound immune dysregulation, manifesting in SLE or susceptibility to Epstein-Barr virus-associated HLH, similar to the clinical phenotype of the Helios R291X patient (supplemental Table  2  and data supplement for detailed clinical information). Importantly, in family D, the Helios R106W variant was also present in 2 family members who were asymptomatic, indicating incomplete penetrance.