Drosophila melanogaster model, we report gain-of-function phenotypes due to misexpression of insect codon-optimized version of human wild-type TDP-43 (CO-TDP-43) using both the binary GAL4/UAS system and direct promoter fusion constructs. The CO-TDP-43 model showed robust tissue specific phenotypes in the adult eye, wing, and bristles in the notum. Compared to non-codon optimized transgenic flies, the CO-TDP-43 flies produced increased amount of high molecular weight protein, exhibited pathogenic phenotypes, and showed cytoplasmic aggregation with both nuclear and cytoplasmic expression of TDP-43. Further characterization of the adult retina showed a disruption in the morphology and function of the photoreceptor neurons with the presence of acidic vacuoles that are characteristic of autophagy. Based on our observations, we propose that TDP-43 has the propensity to form toxic protein aggregates via a gain-of-function mechanism, and such toxic overload leads to activation of protein degradation pathways such as autophagy. The novel codon optimized TDP-43 model is an excellent resource that could be used in genetic screens to identify and better understand the exact disease mechanism of TDP-43 proteinopathies and find potential therapeutic targets.Transactive response DNA binding protein-43 (TDP-43) is known to mediate neurodegeneration associated with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). The exact mechanism by which TDP-43 exerts toxicity in the brains, spinal cord, and lower motor neurons of affected patients remains unclear. In a novel  TARDBP gene in the human genome, has been identified as a major component for the pathology of motor neuron diseases and related neurodegenerative diseases , encoded by diseases . TDP-43 diseases . In the diseases . Mutatiof TDP-43 , and havf TDP-43 . Proteinf TDP-43 . In addif TDP-43 . In ordef TDP-43 .Drosophila melanogaster has been widely utilized to study neurodegenerative diseases in an in vivo model system  defects, and decreased dendritic branching of DA neurons  was added upstream of the start codon for the TDP43 gene. These constructs were subcloned into the Not1-Xba1 site of the modified fly upstream activation sequence (UAS) expression (pEx-UAS) and glass (pEx-gl) vectors . The expression vectors containing the CO-TDP-43 gene were then microinjected into the flies to obtain transgenic flies . We generated multiple transgenic fly lines as noted in UAS-TDP-43_6 line for our subsequent experiments with various GAL4 lines and the gl-CO-TDP-43_4 line for subsequent experiments using the eye as a model. The expression of non-CO-TDP-43 is driven in the fly eye by the glass multimer reporter, GMR-GAL4 on the X-chromosome  (SevEP-GAL4 driver (expressed in R7 and R8 photoreceptor neurons) was recombined with UAS-TDP-43CO to obtain stable transgenic flies expressing w1118;SevEP-GAL4,UAS-TDP-43CO/CyO; +. The GMR-GAL4 on the X-chromosome was placed in trans to the gl-TDP-43CO line to generate GMR-GAL4;gl-TDP-43CO/CyO transgenic flies. The following stocks were obtained from Bloomington Drosophila Stock Center : w1118;UAS-LacZ (BDSC 3955), w1118,GMR-myr-mRFP (BDSC 7121), y1,w1118;Sp/CyO; eGFP-ATG5 (recombined from BDSC 59848), y1,w1118;UAS-GFP-mCherry-ATG8 (BDSC 37749), GMR-GAL4(X) (eye specific) (BDSC 79572), w1118;SevEP-GAL4 (R7 and R8 in photoreceptor cells) (BDSC 5793), w1118,beadexMS1096-GAL4 (wing driver) (BDSC 8860), w1118;Scabrous-GAL4 (sensory organ precursor and wing discs driver) (BDSC 6479), and y1,w*; CCAP-GAL4  (BDSC 25685). Eq-GAL4 (bristle driver) and y1,w1118;Rh1-GAL4/CyO (expressed in R1-R6 photoreceptor cells) were obtained from Dr. Hugo J. Bellen . All crosses were set and flies were maintained at room temperature (22°C) in standard D. melanogaster Jazzmix medium .Codon optimized TDP-43 gene was synthesized from DNA2.0 . The complete sequence of the codon optimized TDP-43 is provided in the romosome . All traAmerica) . SevEP-GAdult retina and imaginal eye discs from third instar larvae were dissected and fixed in 4% paraformaldehyde for 1 h on ice. Adult retina was washed in 0.5% PTX for 3 h to reduce autofluorescence. The tissues were blocked in 0.8% PBS + Triton-X + BSA for 2 h and incubated with primary antibody overnight at 4°C. The tissues were incubated in secondary antibody for 2 h at room temperature, washed in 0.1% PBS + Triton-X and mounted on glass slides with Vectashield . Tissues were stained with the following antibodies: mouse monoclonal anti-TDP-43 antibody , rabbit polyclonal anti-TDP-43 antibody , rat monoclonal anti-Elav , mouse monoclonal anti-GFP , Alexa Fluor 633-conjugated Phalloidin , Alexa Fluor 488 conjugated chicken anti-rat  Alexa Fluor 568 conjugated goat anti-rabbit  and Alexa Fluor 568 conjugated goat anti-mouse .Overexpression of CO-TDP-43 in the fly eye was used to measure total protein levels by immunoblotting. Approximately 50 fly heads were decapitated and homogenized for 1 min in homogenization buffer  along with 1X PhosSTOP phosphatase and 1X cOmplete protease buffer . The homogenized samples were centrifuged at 4 °C for 15 min at 18,000 g. The supernatant was collected and equal parts of the supernatant and Laemmle sample loading buffer with β-mercaptoethanol  was added for each sample. Following a brief pulse centrifugation, samples were loaded on 4%–20% SDS-PAGE gels  for electrophoresis. For higher molecular weight species detection, the fly heads were homogenized in 1X PBS along with the same protease and phosphatase inhibitors. Non-reducing sample loading buffer  was added to the supernatant without β-mercaptoethanol. The blots were blocked in 5% milk, incubated with primary antibodies overnight at 4 °C, washed in 1X TBS + Tween, and incubated with secondary antibody for 1 h at room temperature. The following antibodies were used: Mouse monoclonal anti-TDP-43 antibody , Rabbit Polyclonal anti-TDP43 antibody , Rabbit monoclonal anti-Phospho-TDP43 , Mouse monoclonal anti-tubulin antibody , Mouse monoclonal anti-beta Actin antibody , secondary anti-mouse IgG-HRP , and anti-rabbit IgG-HRP .For Lysotracker staining, imaginal eye discs from the third instar larvae were dissected in 1X PBS solution without fixative. The eye discs were then stained with 100 nM LysoTracker Red DND-99 (Invitrogen) for 2 min, followed by a 1-min wash in 1X PBS. The tissues were mounted on a glass slide with a drop of 1X PBS solution; no Vectashield was added. The coverslip was sealed with nail polish and visualized immediately using a confocal microscope. The z-stack images were analyzed using the ImageJ software .ERG was recorded in 1-day old flies using the same methods as previously described . BrieflyThe adult eye, wing and bristle pictures were taken with a Nikon AZ100 M microscope equipped with a Nikon DS-Fi1 digital camera . Extended depth of focus (EDF) and volumetric images were taken using the Nikon NIS-Elements AR 3.0 software as previously described . The scat-test was performed between the Control vs non-CO-TDP-43 vs CO-TDP-43 eyes for statistical analysis. Western Blot data comparing non-CO-TDP-43 and CO-TDP-43 was estimated by paired t-test. The histograms represent mean ± SEM and are analyzed using Graphpad prism software. Quantification of Lysotracker staining was performed using the NIH Image J software  promoter direct fusion construct specifically generated to study TDP-43 mediated effects on the fly retina (Sevenless (SevEP-GAL4), that only expresses in a subset of photoreceptor neurons (R7 and R8) and cone cells  driver leads to loss of bristles in the adult fly , sca-GAL4, equate-GAL4 (eq-GAL4), and CCAP-GAL4. We found that non-CO-TDP-43 transgene expressed using bxMS1096-GAL4 led to viable adults with shriveled wings, with some flies having wings that were either necrotic or had areas of hyperpigmentation  compared to non-CO-TDP-43 flies  recordings to measure the functionality of active photoreceptor neurons by measuring their response to light stimulus . In factecording . Using tcontrols . The gl-eclosion . These eAn autophagic dysfunction has been implicated in many neurodegenerative diseases, including ALS . TherefoTo further characterize the large vacuoles, we coexpressed CO-TDP-43 and a tagged autophagy protein, Atg5-GFP, which is responsible for the formation of the autophagosomes. We found that these vacuoles were positive for both Atg5 and TDP-43 . During D. melanogaster resource to better understand TDP-43 mediated neurodegeneration. There is great potential for the codon optimized TDP-43 model, as it exhibits robust and sensitive phenotypes ideal for genetic manipulations that allow us to understand its pathogenic mechanisms in an in vivo system. Our results suggest that this model has important utility in understanding the TDP-43 mediated pathology in neurodegenerative disorders.To date, little is known about the exact mechanism of action of TDP-43 mediated toxicity. Here, we report a novel transgenic Previous studies using wild-type TDP-43 fly models helped us understand how the misfolded proteins contributed to neurotoxicity and eventual neurodegeneration . HoweverDrosophila eye can be diverse. For example, in a polyglutamine-expanded human huntingtin transgenic model, the expanded huntingtin protein was shown to form nuclear inclusions and cause severe degeneration of photoreceptor cells (gl-CO-TDP-43 line compared to SevEP-GAL4 driven CO-TDP-43. This is perhaps not surprising considering that SevEP-GAL4 is expressed only in a subset of photoreceptor neurons. Based on our observations using a wide array of eye-specific drivers, we find that TDP-43 pathology is not restricted to photoreceptor neurons but is most likely widespread among different cell types in the Drosophila retina. In fact, TDP-43 is known to be present and shows disease-related pathology across different types of cells both in humans and in animal models (Drosophila model of TDP-43 has been shown to exhibit individual responses in motor neurons and glial cells (In addition, our CO-TDP-43 transgenic model affected multiple different cell types in the fly retina, as evident by depigmentation of pigment cells, irregularities in interommatidial bristle cells, disruption in rhabdomeres morphology, disruption of photoreceptor neuron morphology, and necrosis of the cone cells. The effects of neurodegenerative proteins on the or cells . Furtheror cells . It is wl models . Moreoveal cells . The CO-There is growing evidence of increased levels of TDP-43 protein in the plasma of patients with ALS . More reProtein misfolding is one of the salient features of neurodegenerative diseases. Hyperphosphorylated and aggregated forms of Tau protein are found in the brains of patients with Alzheimer’s Disease . Similarin vitro studies have shown disease-specific mutant or truncated TDP-43 can form toxic aggregates of oligomeric species, very few studies in wild-type TDP-43 animal models have demonstrated a similar robust production of TDP-43 aggregates (CO-TDP-43 induced neurotoxicity was validated by the electrophysiological readouts in our study, which showed that toxic aggregates of wild-type TDP-43 reduced functional activity in photoreceptor neurons in the adult eye. The cytoplasmic mislocalization and presence of toxic TDP-43 aggregates have been well characterized in human patient samples of ALS/FTLD . While igregates . Our stugregates . Previougregates . In keepLastly, in our CO-TDP-43 model, we observed an increase in acidic vacuoles, as evident by lysotracker staining, that are positive for autophagic proteins ATG5 and ATG8 known to be involved in the formation of early and late stage autophagosomes. At this stage our data is too preliminary to infer whether these vacuoles imply an autophagic induction to clear misfolded TDP-43 aggregates or a complete blockage of this process. Autophagic impairment is a common feature of many neurodegenerative diseases, including Alzheimer’s disease, Parkinson’s disease, Huntngton’s disease, and ALS . In patiDrosophila Tauopathy models have shown that misexpression of Tau leads to severe autophagic dysfunction by decreasing lysosomal acidification (All of this evidence strongly suggests that misfolded TDP-43 aggregates might play a role in impairing autophagic processes thereby causing more toxic aggregates to accumulate. Interestingly, our previous studies in fication . FurtherDrosophila TBPH on the codon-optimized human TDP-43 needs to be examined. Several studies have shown that the knock-down of TBPH in fruit flies causes locomotor deficits. The compensation of misexpressed codon-optimized human TDP-43 by endogenous TBPH remains an interesting area of study for a comprehensive understanding of the pathways leading to neurodegeneration. The locomotor and longevity assays following the pan-neuronal expression of CO-TDP-43 is another important area that needs substantial research to open new vistas for developing future therapeutics. Despite these limitations, our codon-optimized model displaying a robust neurodegenerative phenotype compared to the non-codon-optimized transgenics is a promising start. In this study, we have shown that unlike other fly models that rely on pathogenic mutations, CO-TDP-43 model recapitulates many of the known phenotypes of TDP-43 proteinopathies observed in humans.However, our model has a few limitations. A significant aspect to consider for future studies is whether the intrinsic structure and stability of the TDP-43 protein changes due to codon-optimization. There are numerous reports that phosphorylation of TDP-43 results in its mislocalization in the cytoplasm, thus hindering its ability to shuttle back to the nucleus and bind to nucleic acids . Interesin vivo platform to characterize and validate some of these modifiers to better understand the TDP-43-dependent disease mechanism in ALS/FTLD. In conclusion, the robust phenotypes observed in the external organs of the eye, wing, and notum of these flies can be scored easily, offering an excellent model for high-throughput screens of modifiers genes that will help elucidate the molecular mechanism of toxicity due to TDP-43. Targeted genetic screens that identify effectors of TDP-43 will allow us to further identify and pursue novel mechanisms for disease pathology.Furthermore, TDP-43 is an RNA-binding protein that is involved with RNA metabolism and regulation. As a result, much effort has been devoted to identifying the RNA targets of TDP-43 using cell culture models, animal models, and ALS and FTLD patient brain samples. Recently, TDP-43 was shown to bind approximately 30% of the mouse transcriptome, identifying a vast number of possible interactors that can associate with TDP-43 to regulate RNA processing and splicing . Many of


Eucommia ulmoides polysaccharide (EUP), modified SeNP to get EUP-SeNP with a size of about 170 nm, and its effect on 3% dextran sulphate sodium (DSS) induced colitis was explored. Our results showed that colon intestinal histology, intestinal mucosal barrier, inflammatory cytokines and intestinal microbiome composition were changed after EUP-SeNP treatment in colitis mice. Specifically, it was also shown that oral treatment of EUP-SeNP could relieve the degree of DSS-induced colitis in mice by restoring weight loss, reducing disease activity index (DAI), enhancing colon antioxidant capacity and regulating intestinal microbiome composition. In addition, we verified the mechanism in intestinal epithelial cell lines, showing that EUP-SeNP inhibited LPS-induced activation of the TRL-4/NF-κB signaling pathway in intestinal epithelial cell lines. To some extend, our study provides therapeutic reference for the treatment of IBD.Ulcerative colitis (UC) is currently the most common inflammatory bowel disease (IBD). Due to its diverse and complex causes, there is no cure at present, and researchers are constantly exploring new therapies. In recent years, nano-selenium particle(SeNP) has attracted wide attention due to excellent biological activities. Therefore, in this study, for the first time, we used a natural polysaccharide,  Ulcerative colitis (UC) is a chronic, non-specific inflammation of the colon and rectum, mainly manifested as pathological intestinal mucosal injury, ulceration, contraction of the intestine, diarrhea, blood in the stool , 2. It iSelenium(Se), an essential trace element, was thought to be toxic when it was first discovered, and its biological role was gradually discovered . It has In addation, NF-κB is usually related to inflammatory and in the cytoplasm it is inactive but phosphorylated in response to extracellular factor stimulation. Phosphorylated NF-κB can enter the nucleus to bind to target genes and regulate transcription . TLR-4 iIn this study, we used EUP as a surface modifier to modify SeNP to obtain a stable SeNP with anti-inflammatory and antioxidant functions, and then explored its effect on DSS-induced colitis. Finally, we investigated the effect of EUP-SeNP on the LPS-activated TRL-4/NF-κB signaling pathway in intestinal epithelial cell lines so as to provide a reference for the treatment of UC.Eucommia ulmoides polysaccharide   was added to stir for 12 h to obtain EUP-SeNP solution. Excess sodium selite and ascorbate were subsequently removed by dialysis with Mili-Q water in regenerated cellulose tubes  until the Se regenerated cellulose tubes were undetectable by inductively coupled plasma emission spectrometry (ICP-OES) in external solution. After dialysis, some of the reaction products were lyophilized. The remaining liquid is stored at 4 ℃.SeNP was prepared by reacting sodium selenite  with ascorbic acid  in a certain proportion. In brief, 10 mL sodium selenite (20 mM) was added to 10 mL ascorbic acid (80 mM) for mixing and stirring for 1 h under dark environment, and then The Se content in EUP-SeNP was quantitated by ICP-OES on an ICP spectrometer at a specific wavelength of 196.08 nm. The Fourier transform infrared (FTIR) spectra of EUP and SeNP and EUP-SeNP were determined. Briefly, 5 mg of each sample was uniformly mixed with dry KBr solid powder. The powder was sufficiently ground with a mortar and pressed by a vacuum tablet machine. The infrared absorption spectra of these samples were determined on a Nicolet 6700 FTIR (Thermoelectric (Shanghai) Technology Instrument Company, China). The EUP-SeNP were characterized by transmission electron microscope and the energy dispersive X ray spectrometer (TEM-EDX) . A small amount of the sample was placed on the membrane surface of the copper mesh used for electron microscopy. Then, the copper mesh was dried naturally in the oven to avoid the contamination of dust and other impurities. The dried copper mesh was observed with a TEM, and the elemental composition of the composite particles in the target area was observed using a spot sweep method together with an EDX. Particle size was determined by Zetasizer Nano ZS . Vitro antioxidant capacity was evaluated using 1,1-diphenyl-2-picrylhydrazyl(DDPH), hydroxyl radical(·OH), and 2, 2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS) free radical scavenging kit. A total of 60 6–8 week old male C57BL/6JNifdc mice (20-21 g) were purchased from Vital River Laboratory Animal Technology Company . All animals were raised in a controlled environment: relative humidity was 40 ± 10% and temperature was 20 ± 5 ℃, with a 12/12 h light–dark cycle. After 1 week of adaptation, the mice were randomly divided into 6 groups, namely control group, 3%DSS group, 3%DSS + EUP group, 3%DSS + SeNP group, 3%DSS + EUP-SeNP group and 3%DSS + selenite group, each with 10 mice. Except the control group, the other groups were given 3% w/v DSS drinking water for 7 consecutive days, and the control group was given drinking water without DSS. From the 8th day, in the treatment group, 200 µL of the drug was intragastric for 5 days, and the selenium content in the EUP-SeNP treatment group was 0.5 ppm. The control group and DSS group were given the same amount of PBS. Body weight and disease activity index (DAI) scores of mice in each group were observed and recorded daily. The length of the colon was measured when the mice were sacrificed. All animal procedures were approved by the China Agricultural University Institutional Animal Care and Use Committee (AW80212202-2-1).After administration of DSS drinking water, the disease activity index score was recorded every day. The DAI score is a composite score of weight change, stool consistency and stool bleeding. Colon tissues were removed and fixed with 4% paraformaldehyde. The colons were cut into 5 μm sections and stained with hematoxylin & eosin (H&E) and Alcian blue-periodic acid-Schiff (AB-PAS) according to standard procedures.  Histopathological score is a comprehensive measure of inflammatory cell infiltration and changes in tissue damage. The changes in goblet cells in colon tissue were observed by AB-PAS staining. Detailed scoring criteria for DAI and histology are based on previous articles , 29.Proteins were extracted from colon tissues with physiological saline, then detected with a BCA protein analysis kit . IL-6, IL-10, IL-12, IL-17, IL-1β, TNF-ɑ , were determined by an ELISA kit. The results were normalized to the protein concentration of each sample. A microplate reader  was used for detection at 450 nm. Indicators of intestinal antioxidant capacity: Total antioxidant capacity (T-AOC), Superoxide Dismutase (SOD), Catalase (CAT), glutathione peroxidase(GPX), Malondialdehyde(MDA), Glutathione/oxidized Glutathione(GSH/GSSG), myeloperoxidase(MPO) levels were determined by using commercial kits according to the manufacturer’s instructions .Proteins extracted from the colon tissue were separated by 8–12% SDS-PAGE after the concentration was detected by the BCA protein assay kit . They were then transferred to 0.2 μm polyvinylidene fluoride membranes . Membranes were blocked with 5% skim milk for 1.5 h. After blocking, the membranes were incubated with different primary antibodies at 4 ℃ overnight. Claudin 1 , Claudin 3 , Occludin , ZO-1 , TRL-4, IKB, p65, p-IKB, pp65, β-actin. Then, the membranes were washed with Trisbuffered saline Tween (TBST) and then incubated with horseradish peroxidase-conjugated goat anti-mouse IgG, or goat anti-rabbit IgG  for 1.5 h. The membranes were imaged with a Tanon 5200 imaging system. Colon tissue sections were dewaxed with xylene and hydrated in gradient ethanol. Then, the antigen was repaired with 0.01 M sodium citrate buffer. Tissue sections were washed with phosphate buffer  3 times for 5 min each time. Endogenous peroxidase activity was blocked by 3% hydrogen peroxide for 30 min, and non-specific staining was blocked by 5% goat serum for 30 min. Tissue sections were incubated with Ki67 antibody (1/200) and anti-MUC2  at 4 ℃ overnight. On the 2 day, after being washed with PBS, they were incubated with biotin-conjugated goat anti-rabbit IgG  for 2 h and then incubated with horseradish peroxidase (HRP)-streptavidin  for 2 h. DAB chromogenic reagent kit  was used for chromogenic reagent detection. Hematoxylin was used for nuclear re-staining. The primary antibody of the negative control group was replaced with PBS. Images were captured using a microscope .Through the one step TUNEL apoptosis assay kit , terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was used to detect the apoptotic level of colon, IEC-6 and Caco-2. The images were taken with upright DP72 microscope. The contents of the mouse colon were collected and stored in liquid nitrogen. An MN Nucleo Spin 96 Soi DNA extraction kit  was used to extract total bacterial DNA from the sample. The primers were designed according to the conserved region of micro-organism V3 + V4. The primers were used for PCR amplification, and the products were purified, quantified and homogenized to form a sequencing library. The constructed library was first subjected to quality inspection, and the qualified library was sequenced with Novaseq 6000 . Sequence similarity greater than 97% was classified as an operational taxonomic unit (OTU). The OTU composition of different samples was analyzed using principal component analysis (PCA), principal coordinates analysis (PCoA) and non-metric multidimensional scaling (NMDS) based on Bray Curtis analysis. Line discriminant analysis (LDA) effect size, also known as LEfSe, was used to analyze the significance of differences between groups from the phylum to genus level. LEfSe analysis required an LDA score > 4.We evaluated short-term and long-term toxicity by orally administering EUP-SeNP to mice for 7 days and 30 days, respectively, and then detected Aspartate transaminase (AST) and Alanine transaminase (ALT) of serum.  Finally, H&E staining was performed on heart, liver, spleen, lung, kidney and colon.IEC-6 and Caco-2 were cultured in DMEM medium supplemented with 10% FBS, 100 U/mL of penicillin, and 100 µg/mL of streptomycin at 37 °C with 5% CO2 in a humidified atmosphere. Cells were incubated with 50 µg/mL LPS for 36 h, and EUP-SeNP was incubated with 10 µg/mL, 20 µg/mL and 40 µg/mL for pretreatment and post-treatment for 12 h, respectively.IEC-6 was incubated into a 96-well plate, and then incubated for 24 and 48 h by adding different concentrations of EUP-SeNP. Next, the medium was discarded and washed three times with PBS. Finally, 100 µL PBS and 10 µL CCK8  were added to each well and incubated for one hour at 37℃, the optical density was measured at a wavelength of 570 nm. IEC-6 was seeded into a 6-well plate, labeled with fluorescence EUP-SeNP using coumarin-6 as a fluorescence probe, cell nucleus were stained with DAPI  and cell membranes were stained with Dil . Cell uptake was observed within 1 h, 2 h, and 4 h, and photographs were taken with fluorescence microscope.Data analysis was performed with GraphPad Prism . Data are expressed as the means ± standard errors of the mean (SEM). All comparisons of variant parameters between groups were made with one-way analysis of variance, with statistical significance as follows: * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, #P < 0.05, ##P < 0.01, ###P < 0.001, ####P < 0.0001. “*” represents the significance between the control group and the DSS model group; “#” represents the significance between the DSS model group and the treatment group.− 1 to 3397 cm− 1. The C-O-C bond shifts from 1412 cm− 1 to 1383 cm− 1, which indicates that hydroxyl groups are adsorbed on the EUP-SeNP surface. In addition, the peak caused by C-H vibration also changed from 2929 cm− 1 to 2926 cm− 1, which further confirmed the combination of EUP and SeNP. Analysis of the elemental composition of EUP-SeNP using EDX (Fig. The preparation flow chart is shown in Fig. Experimental design as shown in Fig. Low and high power images of colon H&E staining in different treatment groups Fig. A. After DSS-induced colitis is accompanied by a decrease in the number of goblet cells and a decrease in mucin, which was ameliorated by increasing the number of goblet cells and mucin secretion in the treatment group compared with the DSS group Fig. A–B. The TUNEL and Ki67 staining were used to measure the number of apoptotic cells and epithelial cell proliferation, respectively Fig. A–B. The 16 S rRNA gene sequencing was used to detect the colonic microbiota of mice in control, DSS and EUP-SeNP treatment group. OTUs Veen diagram showed that there were 319 common OTUs in the colonic microbiota of mice in each group (Fig. In the control group, there was no significant change in body weight Fig. A and no The different concentrations of EUP-SeNP did not affect the activity of IEC-6 within 24 and 48 h (Fig. IEC-6 protein expression Fig. A and CacDue to the complex and diverse causes of UC, the current treatment is troublesome , 28, 29.Many years ago, Se was widely considered to be biologically inert, but recently, comparing with other forms of Se , researcDSS induced mouse colitis model has been widely used to study animal models of UC because of its similar clinical symptoms to ulcerative colitis , 36. AftSubsequently, We analyzed the colon microbiome and it was noteworthy that although there was no difference in α-diversity analysis among groups, PCoA and NMDS results showed changes in intestinal microbiome composition among groups. And we suspected that this might be due only to the timing of the mold, which also has been seen in previous studies . Our stuFinally, we verified the mechanism of action of EUP-SeNP at the cellular level. LPS is a good inducer of cellular inflammatory models. LPS can destroy intestinal mucosal barrier and reduce the expression of tight junction protein , 49. TheWe prepared a uniform spherical shape EUP-SeNP with a particle size of about 170 nm, which has certain antioxidant function in vitro and in vivo. It can reduce DSS-induced colitis by enhancing intestinal mucosal barrier function, reducing inflammation level, improving antioxidant function, promoting proliferation and reducing apoptotic cells. EUP-SeNP can also regulate the composition of intestinal flora, increase the abundance of beneficial bacteria, reduce the abundance of harmful bacteria and improve colon injury. In addition, EUP-SeNP protects against LPS-induced cell damage by inhibiting the activation of TRL-4/NF-κB signaling.


Stenotrophomonas maltophilia has become one of the major gram-negative pathogens causing nosocomial infections. However, comprehensive analysis of the clinical characteristics and molecular epidemiology of patients with S. maltophilia remains limited.S. maltophilia were collected from April 2019 to March 2022 through the Multi-Drug Resistant organisms clinical research network (MDRnet), consisting of 12 tertiary care hospitals in Japan. The clinical characteristics, outcomes, antimicrobial susceptibility and molecular epidemiology of cases with S. maltophilia colonization and infection were investigated and compared.All patients with a clinical culture growing In total, 78 cases, 44 with colonization and 34 with infection, were included. The median age was 72.5 years (IQR: 61–78), and males accounted for 53 cases (67.9%). The most common comorbidity was localized solid malignancy (38.5%). Almost half of the patients (43.6%) were immunosuppressed, and the most common reason was antineoplastic chemotherapy (30.8%). Respiratory tract was the most common site of colonization (86.4%), whereas bacteremia accounted for more than half of infection cases (55.9%). The overall 30-day all-cause mortality rate was 20.5%, which was significantly higher in infection cases than in colonization cases . Susceptibility rates to ceftazidime, levofloxacin, minocycline, and sulfamethoxazole-trimethoprim were 14.1%, 65.2%, 87.2%, and 100%, respectively. Multilocus sequence typing of the 78 strains revealed that they belonged to 62 different sequence types (STs), of which 42 were known STs and 36 were novel STs.Kaplan-Meier survival analysisS. maltophilia frequently colonizes the respiratory tract, and the mortality is significantly higher in infection cases. Sulfamethoxazole-trimethoprim remains active, but susceptibility to levofloxacin appear to be declining. The strains were genetically highly diverse, consistent with the likely environmental origin.Masahiro Suzuki, PhD, KANTO Chemical co., inc.: Grant/Research Support Sho Saito, MD, PhD, Shionogi & Company, Limited: Grant/Research Support Yasufumi Matsumara, MD, PhD, Beckman Coulter: Grant/Research Support|Presicion System Science: Grant/Research Support|Toyobo: Grant/Research Support David van Duin, MD, PhD, Entasis: Advisor/Consultant|Merck: Advisor/Consultant|Merck: Grant/Research Support|Pfizer: Advisor/Consultant|Pfizer: Honoraria|Qpex: Advisor/Consultant|Roche: Advisor/Consultant|Shionogi: Advisor/Consultant|Shionogi: Grant/Research Support|Union: Advisor/Consultant|Utility: Advisor/Consultant Yohei Doi, MD, PhD, bioMerieux: Advisor/Consultant|FujiFilm: Advisor/Consultant|Gilead: Advisor/Consultant|Gilead: Honoraria|GSK: Advisor/Consultant|Meiji Seika Pharma: Advisor/Consultant|Moderna: Advisor/Consultant|Moderna: Honoraria|MSD: Advisor/Consultant|MSD: Honoraria|Shionogi: Advisor/Consultant|Shionogi: Grant/Research Support|Shionogi: Honoraria


We also analyse the effect of the COVID-19 pandemic on LTx in these populations. (2) Methods: A retrospective, population-based observational study was conducted using the Spanish National Hospital Discharge Database. Multivariable adjustment was conducted with logistic regression to analyse the IHM. (3) Results: We identified 1777 admissions for LTx during the study period, of which 573 (32.2%) were performed in patients with IPF. The number of hospital admissions for LTx rose from 2016 to 2020, both in patients with and without IPF, but a marked reduction was observed from year 2019 to year 2020. Over time, the proportion of single LTx decreased and bilateral LTx increased significantly in both groups. The incidence of LTx complications increased significantly over time along with the increase in the incidence of IPF. No significant differences in the incidence of complications or in the IHM between patients with and without IPF were found. Suffering any complication of the LTx and pulmonary hypertension were conditions positively associated with IHM in patients with and without IPF. The IHM remained stable from 2016 to 2020 in both study populations and was not affected by the COVID pandemic. (4) Conclusions: Patients with IPF account for almost a third of all lung transplants. The number of LTx increased over time in patients with and without IPF, but a marked reduction was observed from 2019 to 2020. Although the proportion of LTx complications increased significantly over time in both groups, the IHM did not change. IPF was not associated with increased complications or IHM after LTx. Lung transplantation (LTx) has been shown to improve the expectations and quality of life of patients with advanced lung disease . The IntLTx can be single or double. In recent years, an increase in the number of double-lung transplants has been detected in the United States and Europe. Although single-LTx means less time on the surgical waiting list, shorter procedure time, and lower risk of complications, double-LTx has shown greater survival and fewer chronic graft-versus-host rejections. Median post-transplant survival is 50% at 5 years ,16,17.The objectives of our study were as follows: (a) to assess the temporal trends in the incidence of LTx in Spain from 2016 to 2020 among subjects suffering or not from IPF; (b) to describe and compare the clinical conditions, such as comorbidities, complications of the transplanted lung, and post-transplant infections of these two populations; (c) to analyse the temporal trends in the length of hospital stay (LOHS) and the in-hospital mortality (IHM) among IPF and non-IPF patients who received a LTx; and (d) to assess whether the COVID 19 pandemic has affected the number or the outcomes of LTx in these patients.We conducted an observational study with data obtained from the Registro de Actividad de Atención Especializada-Conjunto Mínimo Básico de Datos (RAE-CMBD). This is a hospital discharge database that collects information on all admissions to public and private hospitals in Spain . We analThe variables collected by this registry include sex, data of birth, dates of admission and discharge, and if the patient has died in the hospital. Additionally, for each patient, and using the International Classification of Disease 10th version (ICD-10), up to 20 diagnoses and procedures can be recorded. The study population included subjects who had a code for LTx ; LTx were classified as single or bilateral LTx using the ICD-10 codes shown in We grouped admissions by IPF status. We considered IPF patients as those with any ICD10 codes J84.1xx in any diagnostic position, and the rest as non-IPF patients.To assess the presence of comorbidities, we used the Charlson comorbidity index (CCI), using ICD10 codes described by Sundararajan et al. . The CCIProcedures such as haemodialysis, extracorporeal membrane oxygenation, and tracheostomy were identified in any of the procedures fields of the database .The following complications of LTx were considered if recorded in any diagnostic position: lung transplant rejection, lung transplant failure, lung transplant infection, and other or unspecified complications of lung transplant . ComplicIn addition, we evaluated the presence of pneumonia and ventilator-associated pneumonia as complications .Staphylococcus bacteraemia, Gram-negative bacteraemia, Pseudomonas aeruginosa infection, Aspergillus infection, and Cytomegalovirus infection were identified in the database using the corresponding ICD10 diagnosis codes  for continuous variables and total frequencies with proportions for categorical variables. To assess changes over time we used Cochran–Armitage tests for categorical variables, and the test for linear trend or the Jonckheere–Terpstra test for continuous variables. 2 test and continuous variables with the t-test or the Mann–Whitney test, as required.We compared categorical variables using the χWe conducted three multivariable logistic regression models  with IHM as the dependent variable. To construct the multivariable models, the steps were as follows: 1. Bivariate analysis to assess the association of independent variables with the IHM. 2. We selected for the multivariable analysis those variables with a significant association in the bivariate tests, as well as other study variables that have been found relevant in previous investigations. 3. The Wald statistic was used to decide, one by one, if variables were included in the model. Using the likelihood ratio test, consecutive models were compared with the previous ones as new variables were added. 4. For the final model, linearity and interactions between variables in the model was checked to identify which study variables were independently associated with IHM. Odds ratios (ORs) are provided as measure of association along with their 95% confidence interval (95%CI).p value < 0.05 (2-sided) was considered significant.All statistical analysis was performed with Stata version 14 . A The anonymized database of the RAE-CMBD was provided to us by the Spanish Ministry of Health upon request, and after scientifical and ethical evaluation of our study protocol . In SpaiFrom 2016 to 2020, we identified 1777 admissions for LTx in Spain. Over the study period, 573 (32.2%) of all admissions for LTx were performed in IPF-patients. Of these, 291 were single (50.7%) and the remaining bilateral LTx. The number hospital admissions for LTx  in IPF-patients can be seen in p < 0.001); however, bilateral LTx increased significantly in both groups . Over the entire study period, IPF patients underwent more single LTx than non-IPF patients ; however, they underwent less bilateral LTx .The proportion of single LTx decreased significantly over time in patients with and without IPF . p < 0.001). Among the patients who underwent LTx, there were fewer women than men and proportionally fewer women in the IPF group vs. the non-IPF group . Only in non-IPF patients was mean age increased significantly (p = 0.025) between 2016 and 2020.The age ranged from 0 to 72 years for those without IPF and from 13 to 71 years for those with this condition. Over the entire study period, the mean age of IPF patients who underwent LTx was almost 7 years higher than that of those patients without IPF . According to the mean CCI, patients without IPF had more comorbidities than those with IPF (p < 0.001). However, the IHM was similar, 11.5% for IPF and 11.9% for non-IPF patients. Neither LOHS nor IHM changed significantly from 2016 to 2020 in any study group.The total mean LOHS was significantly shorter in the IPF patients (44.7 days) than the non-IPF patients . In the non-IPF group, a significantly increase in the frequency of moderate/severe liver disease and hemiplegia/paraplegia between 2016 and 2020 was observed. However, the prevalence of renal disease and cancer/metastatic solid tumour decreased overtime. The prevalence of rheumatoid disease was significantly higher in the IPF group than in the non-IPF group . Patients without IPF had a significantly higher prevalence of COPD (56.1% vs. 13.1%) and asthma (2.3% vs. 0.9%) than those with IPF. Current tobacco use was not codified in any IPF patient and only three non-IPF. Regarding comorbidities, in patients with IPF, the prevalence of pulmonary hypertension increased over time .As can been seen in Around 25% of the patients undergoing LTx suffered LTx rejections, and another 11% LTx infection. If any possible complications were considered, 51.0% and 49.8% of patients with and without IPF were affected. None of the LTx complications showed significant differences when patients with and without IPF were compared.p = 0.004).The proportion of LTx complications showed a significant increase over time in both groups analysed, specifically lung transplant rejection, lung transplant failure, other/unspecified complications of lung transplant, and any complication of lung transplant. Furthermore, the proportion of lung transplant infection in the IPF group increased significantly, almost four times, between 2016 and 2020 , followed by Pseudomonas aeruginosa (8.5%), and Gram-negative bacteraemia (6.6%). A very similar prevalence was codified among non-IPF patients. Regarding isolated pathogens, those most frequently found among IPF patients were Shown in p < 0.001). Crude IHM in patients with IPF was higher in single LTx, in women, in patients with higher CCI, and in those with any complications of lung transplant; however, all these differences were not significant.IHM was higher in patients with IPF presenting with pulmonary hypertension than those without this condition . IHM in patients with IPF was slightly higher in 2020 (14.0% in 2020 and 12.0% in 2019), but this difference was not significant. No differences were found between 2020 and 2019 in the IHM after stratification by type of procedure, sex, age, CCI, pulmonary hypertension, and any complication of lung transplant.Patients operated on in year 2020 had pulmonary hypertension coded in a higher proportion that those who underwent LTx in year 2019  and pulmonary hypertension  were conditions positively associated with IHM. These two same variables showed a significant association among those without IPF and in the entire population of LTx patients . As founFinally, from 2016 to 2020, and after adjusting by all study variables, IPF was not associated with IHM after LTx .In our study, we identified 1777 patients who received LTx in Spain during the period 2016–2020. Of them, 32.5% were patients with IPF, evidencing an increase in transplants in patients with this disease over time. However, in 2020 there was a marked reduction, both in the group of patients with IPF and in those without IPF. This reduction was due to the SARS-CoV-2 pandemic, and it has also been detected in other series. Thus, in England, the average number of transplants in the first months of the pandemic was 4 per month, while in the months of January and February 2020 it was 12.5 per month . In addiLTx can be single or double, with the advantages of the former being shorter surgical time, less trauma, shorter ischemia time, and shorter waiting time on the surgical list . The eviIn advanced stages, lung diseases are associated with many comorbidities, both respiratory and non-respiratory, which affect the quality of life of patients and may interfere with the transplant prognosis ,34. In oAs in our series, the use of extracorporeal membrane oxygenation is becoming widespread. However, the results obtained in the different series are contradictory and inconclusive. Thus, there are authors who have associated its use with a higher rate of complications , while oAnother complication in our series, which has been multiplied by four from 2016 to 2020, are infections . More specifically, pneumonia occurred in 6.1% of patients with IPF in 2016, compared to 8.1% in 2020. Infections are one of the main post-transplant complications, having been described as the second cause of death. Of these, the most common are respiratory infections, mostly bacterial. In the Spanish series by Aguilar et al., bacterial pneumonia 4 weeks after transplantation occurred in up to 44% of transplanted patients . In our Overall, it has been previously described that the survival of transplanted patients is 89% at 3 months, 80% at one year, and 31% at 10 years. The highest number of reported cases of mortality occur in the first 30 days, due to high rejection or infections, mainly by cytomegalovirus ,49. The In our series, we did not observe an increase in IHM over time in either of the two groups. The overall IHM of patients with IPF was 12.7% in patients with a single-lung transplant versus 11.0% in those with a double-lung transplant. These mortality results are similar to those described in the Spanish series by Prudencio-Ribera et al., who detected a figure of 13.4% in the group of patients with diffuse interstitial lung disease, with IPF being the most frequent disease, detected in 79.6% of patients . It is wEarly IHM in single-LTx has shown contradictory results, with an increase in the series by Arango et al.  and ForcOur study has several limitations. The main one is that the data have been obtained from an administrative database, in which the pathologies and procedures are coded according to the ICD-10-CM. The validity of the RAE-CMBD in identifying LTx has not been assessed; however, studies conducted in the US, using the National Inpatient Sample, concluded that discharge databases are reliable and accurate for national epidemiological investigations . FurtherFuture works, including more detailed clinical and biological data and information on long term mortality after LTx, should be conducted in our country to improve the knowledge and management of patients with IPF.In conclusion, patients with IPF account for almost a third of all lung transplants. The number of LTx increased over time in patients with and without IPF, but a marked reduction was observed from 2019 to 2020. The proportion of single LTx decreased significantly, while bilateral LTx increased significantly among both groups. Although the proportion of LTx complications increased significantly over time, the IHM did not change. IPF was not associated with IHM after LTx.


The Orthopaedic In-Training Examination (OITE) is a multiple-choice examination developed by the American Academy of Orthopaedic Surgeons annually since 1963 to assess orthopaedic residents' knowledge. This study's purpose is to analyze the 2017 to 2021 OITE trauma questions to aid orthopaedic residents preparing for the examination.The 2017 to 2021 OITEs on American Academy of Orthopaedic Surgeons' ResStudy were retrospectively reviewed to identify trauma questions. Question topic, references, and images were analyzed. Two independent reviewers classified each question by taxonomy.Trauma represented 16.6%  of OITE questions. Forty-nine percent of trauma questions included images (100/204), 87.0% (87/100) of which contained radiographs. Each question averaged 2.4 references, of which 94.9% were peer-reviewed articles and 46.8% were published within 5 years of the respective OITE. The most common taxonomic classification was T1 (46.1%), followed by T3 (37.7%) and T2 (16.2%).Journal of Orthopaedic Trauma (24.7%), Journal of the American Academy of Orthopaedic Surgeons (10.1%), and Journal of Bone and Joint Surgery, American Volume (9.3%) remain the most cited sources.Trauma represents a notable portion of the OITE. Prior OITE trauma analyses were published greater than 10 years ago. Since then, there has been an increase in questions with images and requiring higher cognitive processing. The  Trauma is reported to represent approximately 15% of the examination content annually.3The Orthopaedic In-Training Examination (OITE) is a multiple-choice examination developed by the American Academy of Orthopaedic Surgeons (AAOS) annually since 1963 to assess orthopaedic residents' knowledge during training.4-6 For program directors, the OITE is the only national test to objectively assess a resident's orthopaedic knowledge both as a benchmark on the efficacy of the residency education curriculum and on the progress of their individual residents.6 Given the importance of the examination, most residents are motivated to perform well, and most rely on an online learning platform, most commonly ResStudy and Orthobullets, to aid in preparation for the OITE. Only a minority of residents use journal reading as their primary learning material.7The OITE is an important examination for both residents and program directors. For residents, the OITE scores, particularly in the last year of postgraduate training, correlate with the American Board of Orthopaedic Surgery part I scores.8-10 These prior studies analyze questions preceding the conversion to a computer-based examination in 2009, which expanded the opportunity to include more advanced imaging and videos as part of the examination and thus could have influenced the examination content and the types of images included.9 Recently, multiple studies have published an updated analysis of OITE questions in other subspecialties, including pediatrics,11 oncology,12 hand,13 hip and knee reconstruction,14 basic science,15 spine,16 and sports.17 To our knowledge, no update on the analysis of the OITE trauma section has been published.Prior analyses of the trauma section of the OITE were published greater than 10 years ago and studied the OITE between 2004 and 2009.The primary purpose of this study is to determine the representation of trauma questions on the OITE between 2017 and 2021 and to analyze these questions for topic, references, imaging, and question taxonomy. Study results will help inform current orthopaedic residents as they prepare for the annual examination and residency program directors as they develop their program's educational curriculum.18 Taxonomy I questions (T1) assess information recognition and recall. Taxonomy II questions (T2) test data comprehension and interpretation through diagnosis or image interpretation. Taxonomy III questions (T3) test knowledge application for problem solving, such as multistep management or decision making. Chi-squared tests were used to compare categorical data. P < 0.05 was considered statistically significant.The 2017 to 2021 OITE questions published on AAOS's ResStudy online platform were retrospectively reviewed. Questions testing concepts related to musculoskeletal trauma were compiled for analysis. Questions related to traumatic injuries that could otherwise be categorized as spine or hand were excluded. This exclusion criterion aims for the questions analyzed to best reflect the type of injuries that are typically managed by a trauma-focused general orthopaedic surgeon and test orthopaedic trauma principles. By contrast, traumatic injuries to the hand and spine are typically treated by their respective subspecialists, and the authors believe that questions related to these topics are better analyzed in studies of the hand and spine domains of the OITE, respectively. For each question, all associated references, figures, and images were compiled. The percentage of trauma questions for each OITE examination was calculated. The topic of each question was categorized as shoulder and elbow, hip and femur, knee and leg, foot and ankle, pelvis and acetabulum, basic science, and general trauma principles. The number of references for each question was recorded. References were classified by publication year. The year of publication was compared with the year of the OITE, and the time elapsed between the reference publication and the OITE was recorded. For questions with figures, both the number of images included for each question and the imaging modality  were identified. Two independent reviewers classified each question by taxonomy, as previously defined by Buckwalter et al.Trauma questions represented 16.6%  of OITE questions between 2017 and 2021, ranging from 14.0 to 20.9% annually. Topics tested include shoulder and elbow (19.1%), hip and femur (18.6%), knee and leg (12.7%), foot and ankle (14.7%), pelvis and acetabulum (14.2%), basic science (10.8%), and general trauma principles (9.8%), as summarized in Table Images were included in 49.0% 100/204) of questions and 87.0% (87/100) of these contained radiographs, 67.0% of which included radiographs alone, and 20.0% included radiographs in combination with other imaging modalities. Other imaging modalities included were CT (21.0%), clinical images (8.0%), and diagrams (4.0%). Table 04 of queJournal of Orthopaedic Trauma (24.7%), the Journal of the American Academy of Orthopaedic Surgeons (10.1%), and the Journal of Bone and Joint Surgery, American Volume (9.3%). These top three journals account for 44.1% of references. Table On average, there are 2.4 references per question (ranging from 2 to 6). Peer-reviewed journal articles represent 94.9% of all references (ranging from 92.0% to 97.2%), whereas the remaining 5.1% of references were composed of textbooks, websites, and instructional course lectures. The three most common journals cited were the Volume 9.%. These On average, the most common taxonomic classification was T1 (46.1%), followed by T3 (37.7%) and T2 (16.2%). Table 3 but no difference was found compared with prior studies of the OITE that analyzed only questions defined by the AAOS as trauma questions between 2005 and 200910 (P = 0.240). These studies reported that trauma questions accounted for between 18.3% and 19.3% of questions on the examination each year,10 averaging 18.8% of the examination between 2005 and 2009.9 By contrast, another study on 2004 to 2008 OITE questions reported that 28.3% of OITE questions were trauma-related questions,8 which is significantly more than that found in our study (P < 0.001). This difference is likely a reflection of different inclusion and exclusion criteria for trauma questions across studies. Many questions related to traumatic injuries can be categorized as both a trauma question and a question in another orthopaedic subspecialty. For example, a traumatic ankle fracture could be categorized as both a foot and ankle and a trauma question. In our study, we opted to exclude questions related to hand and spine trauma because these injuries are typically managed by their respective subspecialist as opposed to a general orthopaedic surgeon.This study provides an updated analysis of the OITE musculoskeletal trauma section to better inform both residents as they prepare for the annual examination and program directors as they develop their program's educational curriculum. Trauma questions continue to account for a substantial proportion of the examination, representing 16.6% of questions between 2017 and 2021. This is slightly more than the 15% that the AAOS aims for trauma to represent,9 and 30.2%10 of questions. This represents a significant increase in trauma questions incorporating imaging in the past decade . The authors hypothesize that the increased incorporation of images may reflect the transition from a paper-based examination to a computer-based examination in 2009,9 which allows the inclusion of higher quality imaging studies and makes it both easier and less expensive to include images. Of the questions containing images in our study, 87.0% contained radiographs either alone or in combination with other imaging modalities. By contrast, 95% of questions with images contained plain radiographs between 2005 and 2009.10 Residents preparing for the OITE should aim to become proficient in the interpretation of both advanced imaging modalities and plain radiographs, but given the clear emphasis on plain radiographic imaging on the examination, test takers should be aware that plain radiographs are still the most frequently tested.Imaging studies, including plain radiographs, CT, and magnetic resonance imaging, are important in diagnostic and treatment decisions in orthopaedics. This study found that images were included in nearly half of all trauma questions tested. In comparison, prior studies from over a decade ago reported that images were included in only 28.3%2 Le et al2 highlighted that the AAOS recommends references should be published within the past 10 years, which held true for 72.1% of all references cited in our study. The majority (94.9%) of references were primary literature sources, an increase from the 75% that was reported in a prior study of trauma questions.8 The three most frequently cited sources in our study, the Journal of Orthopaedic Trauma, Journal of the American Academy of Orthopaedic Surgeons, and Journal of Bone and Joint Surgery, American Volume, were also among the most frequently cited sources identified in the prior reviews of trauma OITE questions.8-10 Interestingly, only 6% of residents report using journals as their primary study resource, but journals were also rated by the same group of surveyed residents as the most useful for preparation for the OITE.7 In contrast to reading journals, Theismann et al7 report that most residents use Orthobullets (53%), ResStudy (26%), JBJS Clinical Classroom (3%), textbooks (8%), or other resources (4%) in preparation for the OITE. The preference toward utilization of online learning platforms for OITE preparation is supported by program directors, 85% of which recommend the use of Orthobullets and 65.7% recommend the use of ResStudy.19 This highlights that residents should be aware of consistently high-yield journals such as the Journal of Orthopaedic Trauma, Journal of the American Academy of Orthopaedic Surgeons, and Journal of Bone and Joint Surgery, American Volume to supplement the secondary references they often use to prepare for the OITE.OITE question writers are encouraged to cite 2 to 6 high-impact journals for each examination question.18 in 1981, the OITE aims to test the complex cognitive processes necessary for the practice of orthopaedics. Questions are thus designed to fall into one of three different taxonomic classifications based on the cognitive processes required to correctly answer them. Taxonomy 1 questions test recognition and recall of information, taxonomy 2 questions test comprehension and interpretation of information, and taxonomy 3 questions test problem solving.18 Our study found that slightly less than half of questions (46.1%) were T1 questions, which represents a slight decrease from the prior analysis of AAOS-defined trauma questions that reported 60.6%9 and 69.0%10 of questions to be T1 (P < 0.001). The next most frequently tested questions are T3 questions that represent 38% of trauma questions, increased compared with 19.7%9 and 21.6%10 on prior analysis of AAOS-defined trauma questions (P < 0.001). Together, these results suggest that although nearly half of questions still test recognition and recall, there is a slight shift toward inclusion of questions requiring higher levels of cognitive reasoning to correctly answer in our study relative to prior studies of AAOS-defined trauma questions. Residents can focus on case-based practice questions, which are more likely to require higher-order cognitive processing, to adequately prepare for the increased proportion of these questions.20 Of note, there was no difference in the taxonomic distribution of questions that our study reported relative to Seybold et al8's analysis of 2004 to 2008 musculoskeletal trauma OITE questions (P = 0.18), which reported 50.8% T1, 11.0% T2, and 38.2% T3 questions. This study by Seybold et al included both AAOS-defined trauma questions and questions defined by AAOS under different domains, which the authors believed also tested musculoskeletal trauma concepts. The difference in inclusion criteria for this study limits our ability to directly compare it with our findings.As first described by Buckwalter et al8-10 The inability to directly compare the inclusion and exclusion criteria across studies limits our ability to compare our study findings with those reported in previous studies. As the AAOS stopped publishing the complete study guide with the official categorization of all questions in 2016,14 we are unable to compare the questions we categorized under the domain of trauma with those that the AAOS intended to test trauma principles. In addition, our study examines the five most recent OITE examinations that are available on AAOS's ResStudy. Greater understanding of trends in questions may have been possible if we were able to examine all questions since the most recent trauma question analysis in 2009.There are several limitations to our study. The trauma domain is broad and overlaps with nearly all 10 other domains on the OITE. In the design of this study, the authors opted to define trauma questions based on injuries typically treated by a general trauma-focused orthopaedic surgeon and thus excluded questions that could be considered traumatic spine injuries and traumatic hand injuries. Although the authors believe that this definition most accurately reflects the intended domain of question writers, the AAOS does not publish their definition of the musculoskeletal trauma domain for comparison nor do prior studies of the OITE trauma questions.This study highlights that trauma continues to account for a notable proportion of the OITE, which has increased somewhat in recent years. The development of a digital examination has resulted in a substantial increase in the use of images for the trauma questions. Consistent with prior reports, high-impact journals remain notable sources of information for the chosen questions, and questions emphasize higher-level cognitive processes . The results suggest that strategies for residents should include interpretation of images, development of higher-level cognitive processes through case-based practice questions, and awareness of high-impact journals.


Sarcoidosis has been a well-recognised risk factor for pulmonary hypertension (PH) for a long time, but still, the knowledge about this concatenation is incomplete. Sarcoidosis-associated PH (SAPH) is an uncommon but serious complication associated with increased morbidity and mortality among sarcoidosis patients. The real epidemiology of SAPH remains unknown, and its pathomechanisms are not fully explained. Sarcoidosis is a heterogeneous and dynamic condition, and SAPH pathogenesis is believed to be multifactorial. The main roles in SAPH development play: parenchymal lung disease with the destruction of pulmonary vessels, the extrinsic compression of pulmonary vessels by conglomerate masses, lymphadenopathy or fibrosing mediastinitis, pulmonary vasculopathy, LV dysfunction, and portal hypertension. Recently, it has been recommended to individually tailor SAPH management according to the predominant pathomechanism, i.e., SAPH phenotype. Unfortunately, SAPH phenotyping is not a straightforward process. First, there are gaps in our understanding of undergoing processes. Second, the assessment of such a pivotal element as pulmonary vasculature on a microscopic level is non-feasible in SAPH patients antemortem. Finally, SAPH is a dynamic condition, multiple phenotypes usually coexist, and patients can switch between phenotypes during the course of sarcoidosis. In this article, we summarise the basic knowledge of SAPH, describe SAPH phenotypes, and highlight some practical problems related to SAPH phenotyping. Pulmonary hypertension (PH) is a haemodynamic state of elevated blood pressure in the pulmonary artery shared by many conditions, and right heart catheterisation (RHC) remains the gold standard for its diagnosis . For decAnother commonly used PH classification is the World Health Organisation’s (WHO) clinical classification. It consists of five main PH groups in which clinical conditions associated with PH are categorised on the basis of similar PH pathophysiological mechanisms, clinical presentation, haemodynamic characteristics, and management .Sarcoidosis has been long recognised as a risk factor for PH. First, sarcoidosis-associated pulmonary hypertension (SAPH) was simply considered to be secondary to lung disease , but latSarcoidosis is a rare disease, and its incidence and prevalence vary broadly depending on geographic region, race, age, and sex. The highest rates are reported in Northern Europe and in African American individuals, and the lowest in Asia. The estimated incidence in Europe ranges between 1 and 19 cases per 100,000 persons/year ,17. RestIt remains unknown why some patients develop PH and some do not, despite the same demographic and sarcoidosis-related characteristics . The rolSAPH is an independent risk factor for mortality in sarcoidosis and carries up to a 10-fold increase in the risk of death ,26,27,28There are no strong recommendations for SAPH treatment, as evidence from randomised clinical trials (RCT) is limited ,10. GiveUnfortunately, SAPH phenotyping is not a straightforward process. It requires sophisticated diagnostic modalities, and the assessment of some elements, such as vascular remodelling, pulmonary angiitis, or microangiopathy, remains beyond the capability of non-invasive tools. Mathijssen et al. have shared the outcome of their attempt to assess the prevalence of different SAPH phenotypes in a single-centre cohort of 40 patients .Sarcoidosis affects the lungs and thoracic lymph nodes in more than 90% of patients . In the Pulmonary involvement increases the risk of PH almost five times compared to non-pulmonary sarcoidosis. In the population from the Nationwide Inpatient Sample 2016–2018, the prevalence of PH was 19% in cases with pulmonary sarcoidosis and 3.5% with non-pulmonary sarcoidosis; however, there was no information on pulmonary sarcoidosis staging . In publ18F-fluorodeoxyglucose (18F-FDG) uptake was found on positron emission tomography (PET) in the pulmonary artery wall in 19% of 175 sarcoidosis patients, and these patients had higher PAP and PVR compared to patients without the uptake [The ablation, compression, and distortion of pulmonary vessels, together with hypoxic pulmonary vasoconstriction (HPV), are believed to be the first elements of the pre-capillary PH development pathway in the parenchymal phenotype of SAPH by analogy to other chronic lung diseases ,37. HPV e uptake . The meae uptake .The distinctive features of sarcoid inflammation and fibrosis, including a specific lymphangitic fashion of distribution, can be responsible for differences observed between SAPH and PH due to other interstitial lung diseases (ILDs). For example, the same degree of fibrosis is associated with higher PAP in sarcoidosis than in idiopathic pulmonary fibrosis (IPF) and other ILDs ,49. The 1) ≤ 60%pred) or restrictive disease  ≤ 70%pred). They found the parenchymal phenotype in 73% of the patients from their SAPH cohort. Interestingly, not all stage 4 patients with pre-capillary PH were classified as the parenchymal phenotype despite the used definition; 17% of them were assigned to a compression phenotype, and one was diagnosed with chronic thromboembolic PH. Patients with stage 2 pulmonary sarcoidosis also fulfilled the criteria of the parenchymal phenotype. Additionally, one patient with stage 2 disease was categorised as having a parenchymal phenotype despite not fulfilling the criteria, but it was decided that this category was the best fit [A clear definition of a parenchymal SAPH phenotype is lacking. Mathijssen et al. adopted the recommendations regarding PH associated with chronic lung diseases from the latest World Symposium on PH to define the criteria of a parenchymal SAPH. These criteria include pre-capillary PH and moderate to severe pulmonary parenchymal disease; the latter was defined as stage 3 or 4 pulmonary sarcoidosis or severe obstructive  .18F-FDG-PET as a tool predicting response to the treatment is limited and shows not satisfying accuracy [Immunosuppressive treatment was reported to be beneficial in some patients with extrinsic pulmonary vessel compression but ineffective in others ,96,98,99accuracy ,8,96. Puaccuracy ,98; howeaccuracy ,97. It iaccuracy .Mathijssen et al. defined a vasculopathy phenotype as a pre-capillary PH with PVR > 3 Wu, no or mild pulmonary disease, and excluded other causes of PH. Based on that, they classified only 1 patient of the group of 40 as a suspected vasculopathy phenotype . Yet, thImmunosuppressive agents are suggested as the first-line treatment when sarcoidosis vasculopathy is suspected, with the option of the pulmonary vasodilators used in some patients . In patiIt is believed that post-capillary SAPH results from cardiac involvement in the course of sarcoidosis that causes LV function impairment. This hypothetical cause-and-effect relationship has not been validated practically, though no studies addressing this topic can be found. Cardiac involvement has been observed in up to 27% of Caucasian and African American sarcoidosis patients and even up to 80% of Japanese patients; in the majority of them, it was clinically silent ,108,109.In post-capillary SAPH due to cardiac sarcoidosis, glucocorticoids and other immunosuppressive agents are recommended. In addition, the standard treatment for cardiac failure should be applied as appropriate ,10,51. TThe liver involvement can be found in 50–75% of sarcoidosis patients, but symptomatic disease occurs in only 5–15% ,111. PorGlucocorticoids and other immunosuppressive agents are recommended for PH caused by sarcoid liver disease . PulmonaThe true prevalence of SAPH and the prevalence of its phenotypes based on predominant cause remain unknown. In the era of precise and personalised medicine, such phenotyping is recommended to lead the management. However, SAPH phenotyping is not a straightforward process. Apart from lacking criteria and gaps in knowledge, multiple phenotypes usually coexist and patients can switch between phenotypes during the course of sarcoidosis. Sarcoid pulmonary vasculopathy is probably the most under-recognised pathophysiological factor of SAPH development, and post-capillary phenotype seems the least studied. Whether genotypes play a role in SAPH development or SAPH phenotype needs further studies ,112,113.18F-FDG-PET seems to be the most helpful tool to achieve this goal; however, it has not been validated and standardised in this indication. There is a large need for well-planned research projects to properly characterise the epidemiology and phenotypes of SAPH and to establish an evidence-based management for this condition. It is also very important to take sarcoidosis into the differential workup of PH, remembering that it may cause PH without visible involvement of the lungs or thoracic lymph nodes.A transthoracic echocardiogram (TTE) should be performed as an initial assessment in all sarcoidosis patients presenting features indicative of PH ,114,115.


Beyond the acute illness caused by SARS-CoV-2, about one-fifth of infections unpredictably result in long-term persistence of symptoms despite the apparent clearance of infection. Insights into the mechanisms that underlie post-acute sequelae of COVID-19 (PASC) will be critical for the prevention and clinical management of long-term complications of COVID-19. Several hypotheses have been proposed that may account for the development of PASC, including persistence of virus or the dysregulation of immunity. Among the immunological changes noted in PASC, alterations in humoral immunity have been observed in some patient subsets.To begin to determine whether SARS-CoV-2 or other pathogen specific humoral immune responses evolve uniquely in PASC, we performed comprehensive antibody profiling against SARS-CoV-2, a panel of endemic pathogens, and a panel of routine vaccine antigens using Systems Serology in two independent cohorts of patients with pre-existing systemic autoimmune rheumatic disease (SARD) who either developed or did not develop PASC.A distinct qualitative shift observed in Fcγ receptor binding was observed in individuals with PASC. Specifically, individuals with PASC harbored weaker Fcγ receptor binding anti-SARS-CoV-2 antibodies and a significantly stronger Fcγ receptor binding antibody response against endemic Coronavirus OC43. Individuals with PASC, further, generated more avid IgM responses and developed an OC43 S2-specific antibody response with stronger Fcγ-receptor binding, linked to cross reactivity across SARS-CoV-2 and common coronaviruses.These findings from two independent cohorts implicate previous common Coronavirus imprinting as a marker for the development of PASC in individuals with SARDs.Douglas A. Lauffenburger, PhD, Sanofi: Board Member|Sanofi: Honoraria|Sanofi: Honoraria Zachary Wallace, MD, MSc, Bristol-Myers Squibb: Grant/Research Support|Horizon: Advisor/Consultant|MedPace: Advisor/Consultant|Principia/Sanofi: Grant/Research Support|Sanofi: Advisor/Consultant|Shionogi: Advisor/Consultant|Viela Bio: Advisor/Consultant|Zenas BioPharma: Advisor/Consultant Jeffrey Sparks, MD, AbbVie: Advisor/Consultant|Amgen: Advisor/Consultant|Boehringer Ingelheim: Advisor/Consultant|Bristol Myers Squibb: Advisor/Consultant|Bristol Myers Squibb: Grant/Research Support|Gilead: Advisor/Consultant|Inova Diagnostics: Advisor/Consultant|Janssen: Advisor/Consultant|Optum: Advisor/Consultant|Pfizer: Advisor/Consultant Galit Alter, PhD, Leyden Labs: Ownership Interest|Moderna Therapeutics: Employee|Seromyx Systems: Ownership Interest


Previous studies in conversational pragmatics have showed that the information people share with others heavily depends on the confidence they have in the correctness of a candidate answer. At the same time, different social contexts prompt different incentive structures, which set a higher or lower confidence criterion to determine which potential answer to report. In this study, we investigated how the different incentive structures of several types of social contexts and how different levels of knowledge affect the amount of information we are willing to share. Participants answered easy, intermediate, and difficult general-knowledge questions and decided whether they would report or withhold their selected answer in different social contexts: formal vs. informal, that could be either constrained (a context that promotes providing only responses we are certain about) or loose (with an incentive structure that maximizes providing any type of answer). Overall, our results confirmed that social contexts are associated with different incentive structures which affects memory reporting strategies. We also found that the difficulty of the questions is an important factor in conversational pragmatics. Our results highlight the relevance of studying different incentive structures of social contexts to understand the underlying processes of conversational pragmatics, and stress the importance of considering metamemory theories of memory reporting. This measure, along with the respective experimental procedures, was introduced by Notably, there is an experimental measure that combines both report choices and their confidence, and thus allows objective quantitative study of the thresholds that individuals use to base their decisions on: the so-caleld Report-Criterion Probability , showing that they were willing to report information even though they had low confidence in it. Thus, reporting some information  is perceived as more relevant than the accuracy of the retrieved memory. In these contexts, the answering strategy is better described as “loose” and the incentive structure would favor informativeness, even though the overall context is still very formal.As mentioned, previous research studied the reporting strategies in formal and informal contexts , Exp. 3.A parallel case can be made for informal contexts. For instance, a “first date” situation is an example of an informal context likely prompting a constrained answering strategy, because there are no explicit rules and people probably want to only share what they are confident in. In turn, an informal context with a loose answering strategy may be the situation of chatting with friends. Based on these definitions, it is possible to draw some similarities between the incentive structure at a job interview and at a date despite one context being formal and the other informal. In both cases, people would not report a lot of information unless they are reasonably certain about its correctness, that is, they favor accuracy. Likewise, there are similarities between the incentives when testifying in a trial and chatting with friends. In both loose contexts the incentive would be to report maximum information, that is, they both favor informativeness, even though they differ in formality. To verify this suggestion, the present research tested the different patterns of reporting derived from the different incentive structure of each context.Another issue, which has been treated differently in studies on conversational pragmatics, is the difficulty of the material. In some studies, participants had access to the information during the main task  or the iIn the present experiment, we studied the effects of the incentive structure prompted by different social contexts. We manipulated both the formality of the context  and the answering strategy the context elicits  in communicating individual responses to easy, intermediate, and difficult questions. We particularly focused on the decision to report or withhold information in a conversational exchange as a window onto the information the speaker wants to provide. In line with the previous research  recruited on social media took part in the experiment for a small monetary compensation. Most of the participants were in the process of completing their university studies and the rest had completed high-school education. All were native Russian speakers. It is worth to mention that in Russia it is compulsory to pass the Unified State Exam for obtaining the high-school diploma. This guarantees that all of our participants have a similar basic general-knowledge. The sample size was decided on the basis of previous research on the topic (n = 24). Although such sample could in principle be considered limited with respect to the statistical power, the homogeneity of the sample  and the high number of items from different topics (see below) and varying levels of difficulty lead to smaller inter-participant variability than inter-trial variability. In that case, the large number of items used in the experiment increases power  were used in the experiment. Four hundred questions were used in the main experiment and two were used for practice. The questions were selected from a previously normed and validated database of questions . The GKQThe experiment was programmed using Experiment Builder 2.3.38 , and consisted of one practice session with two questions and eight experimental blocks with 50 questions each. The order of the questions in the experimental blocks, the placement of the alternatives on the screen, and the social context for each question were fully counterbalanced. No more than two questions were placed in the same context in a row. All participants had the same practice questions, which were not used in the main experiment.rcP (Report-Criterion Probability) analyses  and Cohen’s avd as measures of effect size. Question difficulty was manipulated a posteriori. Each participant answered 100 questions for each context. To ensure a similar amount of questions per difficulty level, we chose the cut-off points of 0.40 and 0.70. These limits divided all questions in each of the contexts in three levels with approximately one third of the questions per level.Unless otherwise mentioned, we report 2 formality  × 2 answering strategy  × 3 question difficulty  analysis of variance (ANOVA), and pairwise comparisons using the Student’s Our main hypotheses pertained to the proportion of reported answers. However, differences in accuracy and confidence could affect the decision to report or withhold an answer. Thus, we first conducted analyses to check whether accuracy and confidence were similar between conditions. A full report of these analyses is presented in the ez package  with the three variables, formality, answering strategy, and difficulty, using the  package  in R CorIn addition, entering confidence as a covariate also changed the meaning of our measure. As confidence was equated across all conditions, this measure is better interpreted as showing whether participants were conservative or liberal with respect to their own confidence level. Thus, the absolute values of the reporting rates adjusted by the covariate, reported in M = 0.70, SD = 0.15) than in the formal contexts , F = 18.56, p < 0.001, ηp2 = 0.44, and no differences by answering strategy, F = 2.40, p = 0.135, ηp2 = 0.09, or by question difficulty, F = 0.17, p = 0.844, ηp2 = 0.01. This latter result indicates that any effect of question difficulty on actual reporting rates was due to differences in confidence. Descriptive statistics of proportions adjusted by the covariate as well as unadjusted values are presented in F = 3.33, p = 0.044, ηp2 = 0.12, and answering strategy x difficulty, F = 3.95, p = 0.026, ηp2 = 0.14, were both significant.The 3 × 2 × 2 ANCOVA showed a more liberal reporting strategy in the informal  than in the formal context , t(71.9) = 2.98, p = 0.003, avd = 0.18, and also in the loose  than in the constrained context , t(71) = 2.97, p = 0.004, avd = 0.21. For the intermediate questions, participants also applied a more liberal policy in the informal  than in the formal context , t(71.9) = 3.93, p < 0.001, avd = 0.23, and there were no differences for easy questions.To ensure that pairwise comparisons were made over the response means adjusted by the confidence covariate and not over the raw, unadjusted, means, we used the  package  in R CorF = 4.90, p = 0.037, ηp2 = 0.17 and F = 7.11, p = 0.014, ηp2 = 0.23, respectively. Consistent with the more liberal criterion found in the analyses with the covariate, participants reported more answers in informal  than in formal contexts  and in loose  than in constrained contexts . For intermediate questions, participants also reported more answers in the informal  than in the formal context , F = 7.54, p = 0.011, ηp2 = 0.24. For easy questions, none of the main effects were significant, but the interaction was, F = 12.16, p = 0.002, ηp2 = 0.34. The interaction showed that participants reported more answers in the formal loose context than in the formal constrained, t(24) = 3.47, p = 0.002, avd = 0.35, but that they also reported more answers in the informal constrained than in the informal loose context, t(24) = 2.12, p = 0.044, avd = 0.30 . As in the previous analyses, a higher rcP value means that fewer answers are reported and a more conservative strategy is employed, that is, the focus is on accuracy; the opposite is true for lower rcP. We computed rcP for each participant on each of the four social contexts. For completeness, we report rcP for each context and difficulty level in rcP value for the four contexts. When the effect of social context is stronger for a given difficulty level, then there should be more responses falling between the highest and lowest rcP.Using the classical approach to data analysis, the above results showed that the incentives structure of social contexts only affected reporting strategy for intermediate and difficult items. To further verify statistically this conclusion that participants decided whether a response should be reported or withheld based on the confidence threshold, we employed the rcP per participant and difficulty level and entered the results into a one-way ANOVA within-participants, using difficulty as a factor. The results showed significant differences, F = 24.72, p < 0.001, η2 = 0.51. Pairwise comparisons showed that the number of responses between the minimum and maximum rcP was higher for the difficult  and intermediate  questions than for the easy ones , t(24) = 4.49, p < 0.001, avd = 0.95 and t(24) = 6.34, p < 0.001, avd = 1.14, respectively, without differences between difficult and intermediate questions, t(24) = 1.54, p = 0.137, avd = 0.15. In sum, this analysis confirmed that the incentives structure of social contexts has a stronger effect in reporting strategies when answering intermediate and difficult questions than when answering easy questions.Following that logic, we computed the number of responses rated with confidence between the minimum and maximum In this study, we examined context-dependent conversational pragmatics from the perspective of the speaker. In particular, we investigated the effect of the incentive structure subjacent to different social contexts and levels of question difficulty on the amount of information the speakers share. Participants answered general-knowledge questions covering all levels of difficulty  and decided whether to report or withhold their answers in a particular social context. The main results showed that the incentive structure of social contexts, categorized as formal or informal or as constrained or loose, affected reporting rates and thus the amount of information speakers shared in these contexts. That is, the results confirmed our hypotheses regarding the influence of the formality of the context and, crucially, on the strategy that the context elicits. In particular, we expected that in the informal social contexts and those that elicit a loose pattern of answers, participants would favor informativeness and consequently report more information than in the other conditions. This shows that social context influences reporting strategies, thus confirming suggestions made by previous studies of conversational pragmatics .rcP as well as the unadjusted analyses. That is, we analyzed participants’ reported answers separately from confidence in the unadjusted analyses and jointly in the adjusted ANOVA and rc,. PrcP was purposely designed to assess changes in the criterion of memory reporting based on the confidence ratings and thus allows to study threshold changes. Thus, rcP provides stronger evidence than the adjusted ANOVA on the relationship of the reporting strategies implement by participants, confirming the role of confidence in these decisions. In general, the results are highly consistent with the framework of the strategic regulation of memory reporting by The results also showed that the influence of social context vanishes as the certainty regarding the answers’ correctness increases, in particular with easy questions. This supports our third hypothesis regarding the relationship between the level of difficulty of the questions and the incentives structure of social context. We hypothesized that if the question difficulty is more relevant than the social context’s incentives structure, then no differences in memory reporting across social contexts should be found. However, we did find differences for intermediate and difficult answers, supporting the idea that the incentives structure of the social contexts plays a bigger role in memory reporting for these more difficult questions as opposed to answering easy ones. These differences were found in all three types of analyses conducted: the adjusted ANOVA and This research expands and better characterizes the influence of social contexts on memory reporting. Previous research manipulated only one context dimension, formal vs. informal . The resThis research also showed that the strategic regulation of the informativeness-accuracy trade-off can be investigated with general-knowledge questions of all difficulties. Previous research in conversational pragmatics and memory reporting was conducted with difficult questions only  for easy questions social context did not affect reporting, whereas (2) for intermediate questions only formality affected reporting, and, finally, (3) for difficult questions both types of context influenced reporting. These results show that the effect of social context on memory reporting is more complex than previously suggested. When questions are intermediate or difficult, threats to our self-image and the way people present themselves may be more evident, which could increase the perceived strength of the rewards and penalties for focusing on accuracy or informativeness in a given situation. However, for easy questions participants are more prone to ignore the context and its incentive structure, maybe because the chances of a mistake are low and the perceived consequences to self-image are also minimal.We expected that social context may not affect reporting strategies for easy questions because most of them would be rated with high confidence and will thus be reported. However, a similar argument could be made for answers to difficult questions. If they are rated with low confidence, then they should be mostly withheld. From this perspective, only the condition with intermediate questions should be sensitive enough to show the effect of social context. However, this idea was not supported because the results showed an effect of both contexts for difficult questions. One explanation for this is that, as mentioned, the incentives and penalties for focusing on accuracy or informativeness may be perceived as being stronger when questions are difficult. Another explanation is that difficult questions were not difficult enough. Indeed, confidence for difficult questions was near 50%, which suggests that participants did not perceive questions as that much difficult. Arguing for the contrary, however, accuracy for difficult questions was only around 0.25, which is the baseline/chance level in a four-alternative test. If participants were in fact overconfident and considered difficult questions as intermediate, that could explain the effect of social context for these questions. Future research should try to test both of these alternatives to further advance our understanding of the effect of perceived incentives of social situations on memory reporting strategies.Finally, another novel aspect of the present research is the use of multiple-choice task instead of cued-recall or free-recall questions. In previous studies, cued-recall questions were used as a proxy to what a conversation is, that is, a sequence of comments or questions and answers. This notwithstanding, when we are involved in a conversation, sometimes we also provide options along with our questions. Recognition tests are known to be easier than cued-recall tests because they are affected by familiarity , and at While showing novel findings, this study also has some limitations. First, the computerized administration of the questions is not particularly representative of how a natural conversation or interview unfolds. Previous research used different methods to try to mimic conversational exchanges. For example, In line with these limitations, a more naturalistic procedure could have allowed to collect other conversational markers such as silences, hesitations, etc. These markers convey a large amount of valuable information that may help further understand participant’s conversational behavior. Future research on memory reporting would be greatly improved by the inclusion of such conversational markers in the context of a more naturalistic conversation.The aforementioned limitations stem from the selection of a method with a restricted ecological validity. Yet, this particular paradigm and procedure were chosen to be able to isolate the influence of the specific experimental variables. This is particularly important in research lines that are not widely developed and for which it is necessary to set the grounds to build up knowledge.Another important factor to consider is that each individual may react and behave in a particular social context in a rather varied way. Reactions can vary depending on the incentive structure. However, we did not directly manipulate incentives, which was instead done by proxy through the modeled social situations. One possibility to directly manipulate the incentive structure could have been to introduce monetary incentives, but that manipulation would have made the experimental set-up even further removed from a typical conversation situation. In addition, reactions can vary even depending on the emotional, psychological, or cognitive state of the same person. The present research was not intended to study such individual differences and that is why we framed the social contexts with the most prototypical and representative situations. Participants’ responses to prototypical situations inform us about what they perceive to be the incentive structure in these situations and what they think is the most appropriate reporting strategy. Thus, the results presented in this research are informative about general patterns of memory reporting, the perceived expectations in a given social context, and what the usual behavior in response to social challenges could be, and should not be interpreted as indicative of individual responses in particular situations. In line with this, further studies could also use this approach to study communicative behavior of different social and professional groups.Finally, we want to stress that these results were found in a within-participant design; to the best of our knowledge, this procedure has not yet been tested in a between-participant design. There are metamemory phenomena that depend on this important feature. For example, font size only affects judgments of learning in within-participants designs . Thus, iThis study showed the importance of the incentives structure of a given social context for deciding on the best reporting strategy, the one focused on accuracy and reporting fewer answers or the one focused on informativeness and hence reporting more answers. In addition, this research also showed that the difficulty of the materials is relevant for understanding reporting strategies, with easy questions being seemingly less sensitive to variations in social context. This helps advance our understanding of how social situations shape and affect communication flow, in particular how much information people are willing to share. Future research should study other ways people can use to modify their discourse behavior to further reveal the underlying principles guiding human communication.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by ethics committee of the National Research University Higher School of Economics, Moscow. The patients/participants provided their written informed consent to participate in this study.BML and YS acquired funding for research. BML conducted the data collection and completed the initial writing. BML and KL analyzed the data collection. All authors conducted the conceptualization of the research study, and revised and contributed to the final version.This article is an output of a research project implemented as part of the Basic Research Program at the National Research University Higher School of Economics (HSE University) and was carried out using HSE Automated system of non-invasive brain stimulation with the possibility of synchronous registration of brain activity and registration of eye movements.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.https://www.frontiersin.org/articles/10.3389/fpsyg.2023.1004524/full#supplementary-materialThe Supplementary material for this article can be found online at: Click here for additional data file.