On social networking sites, users are continuously exposed to a variety of posts from the networked individuals. Such information may often influence recipients’ perceptions of what is important and goal pursuits such as materialism. Even though several studies have examined the negative consequences of using social networking sites, less attention has been paid to the role of friends’ number and its impact on people’s life goal pursuits. This study aimed to investigate the dark side of online friends and explored why and when more friends in social networking sites would promote materialism. Based on a sample of 264 WeChat users, study 1 discovered that friends’ number positively impacted materialism through extrinsic goal . Additionally, such association was moderated by social comparison orientation and self-esteem. Importantly, self-esteem buffers the detrimental effect of friends’ number on materialism while social comparison orientation increases it. Study 2 further tested the causal relationship and showed that friends’ number on SNS might become a signal to indicate materialism via an experiment. In conclusion, our findings add to the understanding of psychological processes regarding the dark side of online friends’ number and render suggestions for developing positive personal value. Social networking sites (SNSs), such as Facebook, WhatsApp and WeChat, are regarded as a powerful tool for connecting and information sharing. For instance, active Facebook users reached 2.91 billion in the United States ,4. In thFriends in SNSs refer to the connections based on SNSs that share profiles, information and maintain relationship ,8. ClassAlthough recent studies started to focus on some materialistic behaviors by SNSs’ usage such as impulsive buying ,21,22 anTherefore, this paper attempted to explore the impact of SNSs’ friends’ number on materialism by means of a survey of WeChat users and an experiment. The current research contributed to the literature in the following ways. First, prior studies discovered the positive outcomes of friends’ number such as trust and security . We explSNSs provide a convenient venue for social connection and information sharing among online friends . DiffereFriends on social media have some unique properties compared to the traditional offline interactions. On the one hand, social media facilitate communications and interactions among networked friends . More frBesides, some studies explored the negative aspects of number of SNSs’ friends including unfavorable emotional consequences like SNSs’ fatigue –16, mentThis paper argued that friends’ number would lead to high materialism. Materialism can be seen as an internal value concerning the extent to which they believe happiness comes from possessions and acquisitions are the central goal in life, and success in life is measured by material achievement . Holt exH1: The more friends on SNSs, the greater materialism.According to self-determined theory , extrinsFrom displaying self-images to one’s possessions, friends’ posts provide a variety of external cues to activate the external goals ,42. For On the other hand, the majority of information posted by friends is idealized and positively-skewed . BrowsinH2: The effect of number of SNSs’ friends on materialism is mediated by the emphasis on extrinsic goal.We further explored social comparison orientation and self-esteem as two important characteristics that may influence the detrimental effects of friends’ number on materialism. Social comparison orientation refers to individual differences in people’s tendency to compare themselves with others . IndividThis is because first, individuals with high social comparison orientation have high social comparison frequency in SNSs ,50–53. TTherefore, we hypothesized the following:H3: Social comparison orientation moderates the effect of SNSs’ friends’ number on materialism: for consumers with high social comparison orientation, the number of friends is positively associated with the materialism; for consumers with low social comparison orientation, the positive impact of friends’ number on materialism is attenuated.In addition, people high in social comparison orientation are chronically sensitive to what others think about them . GreaterH4: Social comparison orientation moderates the effect of SNSs friends’ number on materialism via extrinsic goal: for consumers with high social comparison orientation, the number of friends is positively associated with extrinsic goal, which subsequently results in high materialism; for consumers with low social comparison orientation, the positive impact of friends’ number on materialism via extrinsic goal is attenuated.Self-esteem constitutes a perception or evaluation of self-worthiness ; that isIn the context of SNSs, for people with high self-esteem, they are less likely to be influenced by friends’ number since they rely less on external information for self-evaluations . It servH5: Self-esteem moderates the effect of SNSs friends’ number on materialism: for consumers with low self-esteem, the number of friends is positively associated with materialism; for consumers with high self-esteem, the positive impact of friends’ number on materialism is attenuated.In addition, according to the previous literature, people with high self-esteem tend to establish their self-worth in the internal contingencies, while those with low self-esteem establish their self-worth in the external contingencies such as appearance and approval from others . On the H6: Self-esteem moderates the mediating route of number of friends on materialism via extrinsic goal: for consumers with low self-esteem, the number of friends is positively associated with extrinsic goal, which subsequently results in high materialism; for consumers with high self-esteem, the positive impact of friends’ number on materialism via extrinsic goal is attenuated.Based on the above theoretical foundations, we derived the theoretical framework for this research in WeChat as the target type of SNS in the current research because WeChat is the most representative SNS among Chinese users, and has been the main data source for studies about SNSs in China ). Hence, hypothesis 2 was supported.To further verify the mediation effect, we conducted a mediation analysis using bootstrapping . Resultβ = -0.64, t = -2.17, p < 0.05), a non-significant main effect of social comparison orientation , and a significant interaction effect , which meant that social comparison orientation served as a moderator to boost the positive impact of friends’ number on materialism. This moderating effect was further verified with bootstrapping .Next, to test the moderating role of social comparison orientation on the relationship between friends’ number and materialism, we conducted a moderated mediation model. The results indicated a significant main effect of friends’ number , compared to people with low levels of social comparison orientation . Specifically, for people with high social comparison orientation, extrinsic goal significantly mediated the relationship between friends’ number and materialism , a non-significant main effect of self-esteem , and a significant interaction effect . The results showed that self-esteem weakened the effect of WeChat friends on materialism .Similarly, we performed a moderated mediation model with friends’ number as independent variable, materialism as dependent variable, extrinsic goal as mediator, and self-esteem as the moderator variable. In support of H4, there was a significant main effect of friends’ number , compared to people with high levels of self-esteem . Specifically, for people with low levels of self-esteem, extrinsic goal significantly mediated the relationship between friends’ number and materialism  factorial between-subjects experimental design. Specifically, we designed the experiment from the recipients’ perspective since as users of the SNS, they know how they were affected by friends, and thus to ensure a positive inferences of materialism in terms of friends’ number.http://www.credamo.com). 160 participants completed the survey in exchange for 2 CNY . 88.8% of the participants were between the ages of 21 and 40.The study was approved by the Institutional Review Board (IRB) of Fujian Agricultural and Forestry University. Participants finished the survey via an online survey platform in China , extrinsic goal . Next, participants were asked to complete the manipulation checks with the following two questions, “to what extent do you think Sunnie’s friends’ number on that SNS is ”, “to what extent do you think Sunnie’s friends’ size on that SNS is ” (α = 0.94). Finally, they provided their demographic information including gender and age.high = 4.69, SD = 1.57; Mlow = 4.10, SD = 1.66; t(158) = -2.31, p < 0.05). This suggested that friends’ number exerts a significant influence on inferences of materialism. Participants thought Sunnie had higher materialism when he had more friends on SNS, which supported the positive relationship between friends’ number and materialism in H1.The analysis of the manipulation check confirmed that participants assigned to the high number condition perceived that the blogger had more friends  than those assigned to the low number condition  = -15.15, p<0.001). Thus, the manipulation of the number of SNS’s friends was successful. Besides, compared to participants in low friends’ number condition, participants who saw a profile with high friends’ number tended to perceive the blogger as more materialism . Besides, materialism was positively associated with extrinsic goal . When we controlled for extrinsic goal, friends’ number was no longer a positive influence on materialism , which suggested a full mediation effect.To test the mediating role of extrinsic goal for the relationship between friends’ number and materialism, we performed a multiple-step mediation analysis . As preds.e. = 0.20, 95% CI = ). The study demonstrated that more friends on SNS led to higher inferences in terms of poster’s extrinsic goal pursuit, which in turn enhanced the extent to which this blogger is perceived as materialism. This could also suggested that for users on SNS, friends’ number serve as a signal of materialism since exposing to more positively-skewed information posted by friends enhanced the extrinsic goal for the users, ultimately leading to high materialism. Hence, hypothesis 2 was supported.To further verify the mediation effect, we conducted a mediation analysis using bootstrapping Click here for additional data file.


A growing body of evidence suggests potential sex differences in the clinical outcomes of COVID-19. In this study we evaluated whether sex and baseline clinical variables were associated with treatment effect after receipt of immunomodulators vs placebo in a study population derived from ACTIV-1 IM clinical trial.ACTIV-1 IM is a master protocol that evaluated infliximab, abatacept and cenicriviroc in addition to standard of care in adult patients hospitalized with moderate or severe SARS-CoV-2 infection through randomized, double-blind comparisons to shared placebo. We performed a subgroup analysis of the modified intention-to-treat cohort. Primary outcome was association of sex with time-to-recovery at day 28, key secondary outcomes were 28-day mortality and clinical status at day 14.A total of 1381 participants were included. There was no difference in treatment effect for primary endpoint of time-to-recovery based on sex for infliximab , abatacept  or cenicriviroc  substudies. There was no difference in treatment effect by sex for secondary outcomes for infliximab and abatacept substudies. The cenicriviroc substudy was terminated early for futility and found no difference in treatment effects based on sex for secondary outcomes either.There was no difference in treatment effect for infliximab, abatacept or cenicriviroc among males vs females. This suggests a more complex interplay of pathogen and host factors deserving further research to identify therapeutic targets for COVID-19.Radica Alicic, MD, MSc, Boehringer Ingelheim Pharmaceuticals, Inc.: Advisor/Consultant Caryn Morse, MD, Gilead Sciences Inc.: Grant/Research Support|Janssen: Grant/Research Support|Laurent Pharma: Grant/Research Support|Moderna: Grant/Research Support|Ridgeback Bio: Grant/Research Support William G. Powderly, MD, Merck: Advisor/Consultant


Rapid and accurate diagnosis of acute respiratory infection (ARI) in children is crucial for proper patient management, and nucleic acid amplification tests are the gold standard. Nasal swabs are preferred for surveillance studies due to their ease of use, mainly in children. Our study compares the performance of two sample-to-answer (STA) platforms for detecting multiple viruses in self-collected nasal swabs from symptomatic children.Between January 2023 and March 2023, self-collected nasal swabs in UTM from children and staff with ARI symptoms were collected as part of an ongoing school-based surveillance study in a large Missouri school district. Samples were tested on the QIAstat-Dx Respiratory SARS-CoV-2 Panel and the fully automated Hologic Panther Fusion system, which can detect a total of 21 respiratory viruses. Percent Agreement analysis was performed to compare the performance of each platform.Of 200 samples collected, QIAstat detected viruses in 111 samples (55.5%) and Hologic in 115 samples (57.5%). Both assays showed high overall agreement (90%), with a PPA (Positive percentage agreement) of 89%, an NPA (Negative percentage agreement) of 94%. Parainfluenza (PIV) showed 100% PPA and NPA followed by adenovirus (Adv)  and rhinovirus (RV) (PPA: 92% and NPA: 94%). Human metapneumovirus (hMPV) had the lowest PPA (47%) but a high NPA (99%) (Table 1). The median CT values for QIAstat and Hologic were 28.8 (IQR 23.65 - 32.5) and 29.3 (IQR 24.2 – 34.5), respectively. QIAstat missed 19 detections, including hMPV , coronavirus NL63 , and RV , with a corresponding median Ct of 39.1 (IQR 34.5 – 41.2) on Hologic. Hologic missed 12 detections, with RV being the most common target , and a corresponding median Ct of 32.9 (IQR: 30.3-35.5) on QIAstat.QIAstat and Hologic exhibit comparable performance  for detecting viral respiratory pathogens from self-collected nasal swabs. Viral detections missed by QIAstat assay had low viral loads, while targets missed by Hologic assay had moderate viral load; additional investigation is needed. Our findings suggest that both STA platforms are suitable for diagnosing respiratory viral infections in self-collected nasal swabs in school-setting.Rangaraj Selvarangan, BVSc, PhD, D(ABMM), FIDSA, FAAM, Abbott: Honoraria|Altona Diagnostics: Grant/Research Support|Baebies Inc: Advisor/Consultant|BioMerieux: Advisor/Consultant|BioMerieux: Grant/Research Support|Bio-Rad: Grant/Research Support|Cepheid: Grant/Research Support|GSK: Advisor/Consultant|Hologic: Grant/Research Support|Lab Simply: Advisor/Consultant|Luminex: Grant/Research Support


T) ventilation. We hypothesized that in a ‘tidal volume challenge’ with a transient increase in tidal volume from 6 to 8 ml Kg− 1, the changes in PVI could predict fluid responsiveness reliably.The plethysmography variability index (PVI) is a non-invasive, real-time, and automated parameter for evaluating fluid responsiveness, but it does not reliably predict fluid responsiveness during low tidal volume  were recorded at baseline VT of 6 ml Kg− 1, 1 min after the VT challenge (8 ml Kg− 1), 1 min after VT 6 ml Kg− 1 reduced back again, and then 5 min after crystalloid fluid bolus 6 ml kg− 1  administered over 10 min. The fluid responders were identified by SVI rise ≥ 10% after the fluid bolus.We performed a prospective interventional study in adult patients undergoing hepatobiliary or pancreatic tumor resections and receiving controlled low V6–8) after increasing VT from 6 to 8 ml Kg− 1 was 0.86 , P < 0.001, 95% sensitivity, 68% specificity, and with best cut-off value of absolute change (ΔPVI6–8) = 2.5%.The area under the receiver operating characteristic curve for PVI value change (ΔPVIIn hepatobiliary and pancreatic surgeries, tidal volume challenge improves the reliability of PVI for predicting fluid responsiveness and changes in PVI values obtained after tidal volume challenge are comparable to the changes in SVI. T) values, could significantly influence PVI values ; (X = 50 for men and = 45.5 for women). GE Avance® CS 2 anesthesia machines .On the day of surgery, intravenous access was obtained. Demographic and anthropometric data including age, sex, height, actual body weight (ABW), PBW, body mass index (BMI), body surface area (BSA), smoking history, comorbid diseases, and use of vasopressors were recorded. All patients were monitored using 5-lead electrocardiography, non-invasive blood pressure monitoring, and pulse oximetry. General anesthesia was induced with propofol  was greased with a lubricating gel and passed nasally into the mid-esophagus until the aortic blood flow signals were best identified. The Masimo Pulse Co-Oximeter probe  was placed on the index finger of patients and covered to avoid light interference. The Stroke volume (SV) and stroke volume variation (SVV) were measured directly by the trans-esophageal doppler probe. The stroke volume index (SVI) was calculated by dividing the value of SV by BSA . The PVI and Perfusion index (PI) values were reported by the aid of The Masimo Pulse Co-Oximeter monitor. Throughout surgery, packed red blood cells (300 ml) were transfused when the hematocrit level was < 25%. Fresh frozen plasma (200 ml) was administrated when the fibrinogen level was < 2 g dl− 1 or the international normalized ratio was > 2. Patients were extubated either in the operating room or postoperatively in the ICU after completion of surgical intervention.A 7 F triple-lumen central venous catheter was inserted in the right internal jugular vein with real-time ultrasound guidance and complete aseptic precaution. A pressure transducer was connected to the 16-gauge lumine to continuously monitor the central venous pressure (CVP). The left radial artery was cannulated for measuring mean arterial pressure (MAP) invasively. The pressure transducers for both CVP and invasive arterial blood pressure were placed on the midaxillary line, fixed to the operating table to keep the sensor at the atrial level, and zeroed to atmospheric pressure. An indwelling urinary bladder catheter was inserted to monitor urinary output. Head and extremity wrap and forced warming system were applied to maintain body temperature. A trans-esophageal doppler probe (− 1 VT ventilation (baseline 1 time-point) were recorded. After baseline measurement, VT was increased to 8 ml kg− 1 PBW for 1 min , and the hemodynamic variables were obtained. The VT has been reduced back to 6 ml kg− 1 PBW again and after 1 min, the hemodynamic measurements were recorded (baseline 2 time-point). After these hemodynamic measurements, volume expansion was performed for 10 min using an infusion of a balanced crystalloid solution (6 ml kg− 1 ABW). The same hemodynamic parameters were measured under ventilation with a VT of 6 ml kg− 1 five minutes after fluid bolus administration (after fluid bolus time-point). Then, the absolute change (ΔPVI6–8)  and percentage change (%ΔPVI6–8)  between the PVI at 6 ml kg− 1 of PBW (PVI6) and that at 8 ml kg− 1 of PBW (PVI8)  were calculated. The percentage change in SVI, according to volume loading, was used as the principal indicator of fluid responsiveness. Responders or non-responders were determined when the increase in SVI was ≥ 10% or < 10% after fluid bolus, respectively , or percentage (%). Distribution normality was assessed using the Shapiro-Wilk test. The changes in continuous variables from 6 to 8 mg kg− 1 PBW  and the changes before and after fluid bolus were compared using the paired t-test or Wilcoxon signed rank-sum test. Group comparisons between responders and non-responders were performed using the independent t-test or Mann–Whitney U test. Categorical variables were analyzed using the chi-square test or Fisher exact test when indicated. To test the abilities of the dynamic preload indices to predict fluid responsiveness, the AUCs of responders were calculated and compared using the Hanley–McNeil test   9.5% [9–12 (4–29)] at VT 6 ml Kg− 1 baseline 1 time-point to 18% [12–19 (7–35)] at VT 8 ml Kg− 1 after VT challenge, P < 0.001. This difference was not significant in non-responders. , 0.33–0.66), P = 0.99 showing non-significant prediction of fluid responsiveness. The AUC of PVI8 was 0.79 , P = 0.001 showing a sensitivity of 85% and a specificity of 61% for a best cutoff value of 11.5%. The AUC of ΔPVI6–8 was 0.86 , P < 0.001 showing a sensitivity of 95% and a specificity of 68% for a best cutoff value of a 2.5%.  were within the inconclusive grey-zone range of the test (1–6%). The fluid bolus cause PVI values reduction in fluid responders from median [IQR (range)] 11.5 [9.2–16 (5–29)] at VT 6 ml Kg− 1 baseline 2 time-point to 9 [5.5–10 (3–17)] at VT 6 ml Kg− 1 after fluid bolus, P < 0.001. This difference was not significant in non-responders, P = 0.12. The differences between fluid responders and non-responders were comparable at the same time-points.  reliably predicts fluid responsiveness with a cutoff value of 2.5% with excellent prediction of most of fluid responders, 95% sensitivity and with uncertain prediction of fluid non-responders (could miss one in every three fluid non-responders) (30% of non-responders recognized imperfectly as fluid responders), specificity 68%. The PVI6 has no potential to predict fluid responsiveness efficiently and the inconclusive grey-zone range of ΔPVI6–8 lies between 1% and 6% indicate uncertain predictive value. It is a quite wide range, but the total number of patients involved within the inconclusive grey-zone was small. There is associated intrathoracic pressure rise with providing mechanical ventilation using higher VT of 8 ml kg− 1. The subsequent cardiopulmonary interactions significantly lower the cardiac preload status in certain groups of patients (fluid responders). This effect is masked when using a low VT ventilation that minimizes these cardiopulmonary interactions and makes it difficult to differentiate between the fluid responders and non-responders. Transient increase of VT during mechanical ventilation uncover this effect and help to identify the fluid responders [T 8–10 ml Kg− 1 continually throughout data acquisition time and they found PVI was reduced from 14 to 9% after 500 ml hetastarch 6% fluid bolus. They determined a cutoff value of PVI = 14% for fluid responsiveness with 81% sensitivity and 100% specificity. They highlighted a significant relationship between PVI values change and cardiac index variations after the fluid bolus. They waited equilibrium for four minutes (3 min after volume expansion and one minute after no stimulation) after fluid bolus to collect the hemodynamic variables to assess the effects of volume expansion on cardiac output and other hemodynamic variables [T ≥ 8 ml kg− 1 as compared with 6 ml Kg− 1 in prediction of hemodynamic instability before applying sudden positive PEEP value of 10 cmH2O with acceptable sensitivity and specificity in a group of patients who were sedated and mechanically ventilated postoperatively [T ventilation (6 ml kg− 1) could not predict the hemodynamic instability efficiently following sudden PEEP application [− 1 VT for mechanical ventilation after anesthesia induction to compare three pulse oximeter sensors sites . They reported that the three sensor sites were able to predict the fluid responsiveness efficiently and comparably between the three sites [T challenge during mechanical ventilation under anesthesia was used efficiently to improve the capabilities of the dynamic indices for fluid responsiveness prediction [T challenge using 8 ml Kg− 1 VT could increase the predictability of the pulse pressure variation (PPV) and SVV for fluid responsiveness with favorable sensitivity and specificity to avoid unnecessary intravenous fluid administration of such critical group of patients [T challenge could not improve the reliability of SVV to predict fluid responsiveness as there were insignificant differences between the responders and non-responders at all time-points of assessment in addition to that all comparisons before, during, and after VT challenge were comparable. This discrepancy could be attributed to the different methods of measurement of SVV or different cohort of patients between the current trial and that trials of Messina and colleagues [T challenge 8 ml Kg− 1 with sensitivity 75% and specificity 76% in a group of ICU patients who were sedated, mechanically ventilated, diagnosed with septic shock, and with possibly defective lung compliance. They obtained SVV values by transpulmonary thermodilution method [T challenge, before and after fluid bolus, and between fluid responders and non-responders at all time points of assessment. These findings emphasized the poor ability of these hemodynamic variables to differentiate between fluid responders and non-responders. There was statistically significant difference for CVP values before and after the fluid bolus for the fluid non-responders only but of minute clinical effect. This difference may highlight that an increase in preload in fluid non-responders will not augment cardiac output and could impair the cardiac function by increasing the preload of right ventricle. CVP values before and after VT challenge for both fluid responders and non-responders were comparable and the CVP values before and after fluid bolus for responders were comparable denoting the inability of this parameter to predict fluid responsiveness efficiently. The current study utilized the independent SVI variables as a cardiac output surrogate to differentiate between fluid responders and non-responders. It is calculated by dividing the SV value  by BSA. It is not as accurate as the values obtained by the more invasive pulmonary artery catheter with standard thermodilution method [r2). The radius of the descending aorta  is derived from published nomograms based on age, sex, weight, and height  [T challenge for both fluid responders and non-responders were comparable. On the other hand, there were statistically significant differences of PI values before and after fluid bolus for both the fluid responders, P = 0.03 and non-responders, P = 0.01. This effect could be interpreted as PI is not a good predictor of fluid responsiveness, but fluid bolus administration could increase the values of PI in the fluid responders and non-responders similarly without consequent impact on the cardiac output. The PI value changes may possibly suggest intravascular volume status changes that not based on respiratory alterations and cardiopulmonary interactions. These findings go in line with the findings of Cannesson and colleagues [This study reveals the ability of Vsponders . The inhsponders , 21 decrsponders  might beariables . The finratively . They delication . In the ee sites . The VT ediction , 13. Mespatients . In contlleagues , 13. In n method . The valn method . but pron method . or valun method . The mincal.com) . The rescal.com) –30. The cal.com) . The vallleagues .The current trial is a non-randomized study recruiting 64 successive patients for hepatobiliary and pancreatic tumor resection. Its findings should be manipulated and interpreted carefully as data acquisition done under rigorous controlled circumstances in selected group of patients. We did unplanned additional patients’ exclusion before and after enrolment process to keep this non-randomized cohort of patients homogenous under same standard circumstances to minimize the risk of bias. Before enrollment, one laparoscopic hepatobiliary tumor resection case was excluded as the pneumoperitoneum could increase intraabdominal pressure with subsequent affection of respiratory alteration and heart-lung interactions that could interfere with PVI values changes and accuracy of fluid responsiveness prediction. Tissue dissection, third space fluid losses, fluid shifts, and intraoperative blood loss were supposed to be less in laparoscopic surgery than in open type procedures. An additional factor to exclude this case is frequent position changing (head up and down positions). There were four cases of irresectable tumors, and one case of undiagnosed metastasis that were excluded from the study after enrolment as these inoperable cases were subjected to less negligible tissue dissection, fluid shifts, and blood loss than the patients undergone complete tumor resection who may require more volume expansion, blood transfusion, or vasoactive drugs administrations. There was one case of massive blood loss excluded after enrollment as the patient needed massive blood transfusion with high dose vasopressors and experienced marked hemodynamic instability that precluded protocol completion. The five minutes interval between completion of crystalloid fluid bolus (over ten minutes) and acquisition of hemodynamic variable after fluid bolus was relatively long. This delay might result in fading and possibly underestimating the assumed clinical effects of volume expansion and the final outcome.T challenge improves the prediction accuracy of PVI to reliably identify fluid responders and non-responders as changes in PVI values obtained by transient increase of VT to 8 ml Kg− 1 are more predictive than PVI values that are reported during low tidal volume ventilation (6 ml Kg− 1).For optimizing intraoperative fluid administration in hepatobiliary and pancreatic surgeries, V


Precision radiotherapy refers to the ability to deliver radiation doses with sub-millimetre accuracy. It does not however consider individual variation in tumour or normal tissue response, failing to maximise tumour control and minimise toxicity. Combining precise delivery with personalised dosing, through analysis of cell-free DNA, would redefine precision in radiotherapy. Around 50% of cancer patients require radiotherapy at some point in their treatment. The probability of treatment success is dependent upon the dose delivered and the relative radio-sensitivity of the tumour. The maximum dose delivered is calculated to ensure that no more than 10% of patients suffer life-changing late radiotherapy toxicity with an additional 20% reporting toxicity that still has some impact on quality of life. The current “one dose fits all” approach to radiotherapy treatment contrasts with the ethos of precision oncology in other branches of cancer medicine, which increasingly harness the power of genetic and genomic analyses to stratify patient care, monitor tumour response and detect disease progression.Liquid biopsy - the analysis of cell-free DNA (cfDNA) and other cellular components released from dying or damaged cells into the circulation or other bodily fluids - has utility because the cellular components carry the characteristics of the tissues from which they are derived. Circulating tumour-derived DNA (ctDNA) retains the genetic and epigenetic hallmarks of the originating tumour, including somatic mutation and DNA methylation. Attractive because it is minimally invasive and repeatable, liquid biopsy is gaining traction in early detection and diagnosis, patient stratification, detection of minimal residual disease and prediction of recurrence after primary treatment –5.Analysis of cfDNA has been little applied to management of radiotherapy, especially dosing or scheduling. However, the ability to quantify ctDNA and to define cfDNA tissue-of-origin in real time have the potential both to permit assessment of tumour response to radiation during a course of radiotherapy treatment and, at the same time, to measure the extent of damage to healthy tissues surrounding the targeted tumour. These advances, technically possible, would allow truly personalised adjustment to radiotherapy dose per fraction or number of fractions, based on how the patient is responding to their treatment and would transform how radiotherapy is currently delivered.Here, we review recent data that define the mechanism and timing of cfDNA release arising from tissue damage and cell death after exposure to ionising radiation and the extent to which this can be quantified and ascribed to tissue-of-origin. Finally, we consider the work still required to validate and clinically translate these observations to new precision in radiation therapy.Cellular DNA is released into the circulation primarily from dividing, damaged or dying cells undergoing apoptosis or necrosis. In radiotherapy, ionising radiation causes direct and indirect DNA damage predominantly through single and double strand breaks . If thisIn one of the first studies investigating the kinetics of ctDNA release during radiotherapy, Lo et al. quantified Epstein-Barr virus (EBV) ctDNA in patients undergoing radiotherapy for nasopharyngeal cancer. Having observed that 2 out of 10 patients had a transient rise in circulating EBV DNA one week after initiation of radiotherapy, a further 5 patients were recruited, in whom EBV DNA levels were measured daily during the first week. All 5 patients exhibited a transient rise in EBV DNA in the first week of radiotherapy, followed by a fall . More reThese studies suggest a pattern of release of ctDNA from tumour cells, in which damaged cells undergo a combination of immediate and delayed cell death, with an initial rise of ctDNA release into the circulation following exposure to radiation, and detectable ctDNA concentrations that subsequently fall as tumour bulk decreases.ctDNA in cancer patients usually constitutes a small, frequently tiny proportion of the total circulating cfDNA. In early-stage cancer, ctDNA generally accounts for less than 1% of total cfDNA and only in late-stage cancer and in a subset of high-secreting tumours, does the proportion of ctDNA rise above 1% –12. MethTumour-derived somatic mutation in cfDNA can be detected by targeted or genome-wide sequencing and by multiplex and allele-specific PCR. A range of methods have been developed to maximise sensitivity and specificity including barcoding, amplification, error correction and deep sequencing –13. SuchIn virus-associated cancers, detection of viral cfDNA by droplet digital PCR (ddPCR) or sequencing has been used to screen for virus-associated tumours in healthy populations and for characterisation of viral status at diagnosis. In longitudinally collected samples post-treatment, cfDNA viral titre has been successfully used as a prognostic marker and early marker of disease progression. Such approaches have shown particular value in EBV-associated nasopharyngeal cancer and human papillomavirus (HPV)-associated oropharyngeal cancers –24.Advances in radiotherapy technology have improved the accuracy of radiation delivery to the tumour and clinical trials have refined dosing schedules with resultant improvements in outcomes, as reviewed elsewhere , 25, 26.In EBV-associated nasopharyngeal cancer, Lo et al. defined the kinetics of ctDNA release through measurement of EBV cfDNA , leadingOf perhaps even greater novelty and applicability for radiotherapy would be the development of biomarkers of radiation-induced damage to tissues surrounding the targeted tumour. In the past five years, it has become clear that DNA methylation analysis has this capability. Extensive methylation atlases of healthy tissues and tumour types have been developed with the power to define the tissue composition of cfDNA samples and the ability to detect with high sensitivity the presence of DNA from a wide range of tissues and tumours, with demonstrated or potential application in the context of autoimmune disease, transplant rejection and early stage cancer , 18–20. These proof-of-concept studies offer the prospect of direct tests both for tumour cell death and off-target tissue damage during radiotherapy treatments, as has also been noted in other recent reviews , 36. SucWhile evidence for the value of ctDNA analysis following primary treatment with surgery, chemotherapy and radiotherapy is accumulating rapidly, the use of cfDNA analysis to guide radiotherapy dosing and scheduling during treatment is at an early stage. The existing proof-of-concept studies need confirmation in further clinical studies for example on the kinetics of cfDNA release in different tumour types and with different radiotherapy protocols. Technical advances are also required, such as development of protocols to measure DNA methylation directly in real-time, as may now be achievable by single molecule sequencing –39. Howe


To determine whether human hydrosalpinx fluid might have a deleterious effect on thefertilization rate and embryonic development of the exposed mouse oocytes.Mouse cumulus-oocyte complexes (COCs) were randomly allocated for exposure to purehydrosalpinx fluid , EBSS containing 50% of hydrosalpinx fluid and pure EBSS .p=0.031). Theblastocyst formation rate was also lower in the 100% HSF group than 50% HSF and thecontrol group , but not statisticallysignificant (p=0.275). There was no significant difference in the meannumbers of cells in the ICM, TE, and total cell number in blastocysts from the controlgroup and two hydrosalpinx fluid exposure groups.The results showed that the fertilization rate in the 100% HSF group was significantlylower than the control group (64.0% versus 73.0%, Human hydrosalpinx fluid has a negative effect on the fertilization rate of the exposedmouse oocytes. However, this effect was found only in undiluted concentration and doesnot affect the subsequence of embryonic development and blastocyst cell number. In addition to infertility, the presence of hydrosalpinx has adverse effects on assistedreproductive technology (ART) outcomes  was collected in an aseptic manner from hydrosalpinx of sixpatients undergoing laparoscopic surgery for infertility. After the presence ofhydrosalpinx was confirmed laparoscopically, a sterile needle was used to aspiratehydrosalpinx fluid through the fallopian tube lumen. The hydrosalpinx fluid specimen wasimmediately stored at-70°C until used. Before use, hydrosalpinx fluid was warmed to 37°C,and then 2 ml of hydrosalpinx fluid was used for routine bacterial culture, pH andosmolarity check. The pH valued was determined by a pH meter . The osmolarity was determined by a digital osmometer . The remaining hydrosalpinx fluid was usedin the experiment. Mouse embryo assay was repeated six times, each time using hydrosalpinxsample from one patient.Outbreed female and male International Cancer Research (ICR) mice were obtained from theNational Animal Institute, Mahidol University, Bangkok, Thailand. They were cared for atAnimal Husbandry Unit, Faculty of Medicine, Chiang Mai University. All procedures relatedto mice followed the international and national guidelines for ethical conduct in the careand use of animals for research. The room was adequate ventilation at 25±2°C, underhumidity of 60-70%, and controlled 12-hour light/12-hour dark cycles. The mice wererested, not disturbed for seven days before the experiment to avoid the effect of stressfrom transportation. The Animal Ethics Committees of the Faculty of Medicine, Chiang MaiUniversity approved the use of mice in our study under approval no. 28/2563.Five- to nine-week-old female mice were super-ovulated by an intraperitoneal (IP)injection of 10 IU pregnant mare serum gonadotropin ,followed 48 hours later by an IP injection of 10 IU human chorionic gonadotropin . Sixteen hours after the second injection, the mice werekilled by cervical vertebrae dislocation. The peritoneal cavity was exposed, and the twooviducts were aseptically removed and placed in Earle’s Balanced Salts Solution , containing 0.5% bovine serum albumin. Cumulus-oocyte complexes (COCs) were removed from the oviductand separated into three groups for the experiment.2at 37°C.To assess the effect of hydrosalpinx fluid on oocytes, two experimental groups and acontrol group were studied;100% HSF group: pure hydrosalpinx fluid, 50% HSF group: EBSScontaining 50% of hydrosalpinx fluid, and control group: pure EBSS. The COCs were exposedto the assigned condition for five minutes. Following the exposure, COCs were washed inEBSS and transferred to 50 µl drops of fertilization medium  under mineral oil  and use for IVF. All steps were donein an IVF chamber  under an atmosphere of 6% CO2, 5% O2,and 89% N2 at 37°C. The spermatozoa were transferred to COCs drops forinsemination at a final motile sperm concentration of 2.5x105 /ml. Allexperiment groups from the same hydrosalpinx sample were used spermatozoa obtained fromthe same male. Two hours later, MII oocytes were transferred to culture in 10 µldrop of cleavage medium  under mineral oil(IrvineScientific). The fertilization rate was determined the next day by counting thenumber of two-cell embryos. Seventy-two hours post insemination, the embryos weretransferred to blastocyst medium  under mineral oil(Irvine Scientific) and cultured in similar conditions for 48 hours. Embryo developmentwas evaluated under an inverted microscope every 24 hours until completion of 120 hours.Mouse blastocysts were classified as early, partial, full, expanding, hatching and hatchedblastocysts, using the criteria proposed by Male ICR mice aged 10-12 weeks were killed by cervical vertebrae dislocation. Both caudaepididymis was removed and placed in 1 ml of fertilization medium (Cook). Capacitation wasallowed to proceed for 30 minutes in an atmosphere of 6% CODifferential staining was performed on all expanding, hatching, and hatched blastocysts,using the protocol described by p<0.05 wasconsidered statistically significant.Statistical analysis was performed using SPSS program version 16. The Chi-square test inR × C list data was used to compare fertilization rate and blastocyst formationrate in three groups. The mean numbers of ICM and TE cells were compared by one-wayanalysis of variance (ANOVA) when data distribution was normal or the Kruskal Wallis testwhen normality could not be confirmed. A two-tailed versus 73.0%,p=0.031, versus 73.0%, p=0.394). The blastocyst formation ratewas lower in the 100% HSF group than the 50% HSF and the control group , but notstatistically significant , TE , and total cell number in blastocysts from the control group andtwo HSF exposure groups (p=0.479).There was no significant difference in the mean numbers of cells in the ICM , whereas the COCs exposed toother hydrosalpinx fluid samples did not affect the fertilization rate . Furthermore, we continued to observe and found that theblastocyst formation rates and their cell number are also comparable. This the first reported study on the effect ofpure HSF on the exposed oocytes. The results show a significantly lower fertilization rateof oocytes exposed to pure HSF for five minutes before IVF compared to the control , and found a tendency of lowerblastocyst formation rate although not statistically significant. Therefore, hydrosalpinx fluid may have a directcytotoxic effect on oocytes, and this effect can be obscured by the effect of thedilution.This study was designed to use the mouse model to assess the influence of humanhydrosalpinx fluid on the ability of oocytes to fertilized and subsequence embryonicdevelopment. We designed to exposed oocytes to hydrosalpinx fluid for five minutes. Thismethodology mimics the real condition in clinical practice that accidental exposure ofoocytes to hydrosalpinx fluid during oocyte retrieval procedure was not longer than fiveminutes. Likewise, in terms of the hydrosalpinx fluid concentration, we designed to exposethe oocytes with both pure hydrosalpinx fluid to see the real effect, and 50% dilutedhydrosalpinx fluid mimics the real condition in which the follicular fluid from theretrieved oocyte will dilute the effect of the hydrosalpinx fluid. The current evidencesupports that hydrosalpinx reduces the IVF success rate, but the exact mechanism remainsunclear. One proposes mechanism is a direct cytotoxic effect on embryos from the compositionof inflammatory substances, microorganisms and free radicals  impaired the essential nutrients for the cytoplasmicmaturation of the oocytes. Hydrosalpinx fluid contains a composition similar to those inserum but the steroid hormones, glucose, and proteins that necessary for development arelower than follicular fluid (Further analysis on the effect of hydrosalpinx fluid on each oocyte was conductedindependently. We found that only one out of six human hydrosalpinx fluid samples have adeleterious effect on the fertilization rate of the exposed mouse oocytes, while the otherfive samples were not affected. Human hydrosalpinx fluid comprises a heterogenous naturewith an individual variation in the components such as electrolytes, cytokines, nutrients,as well as endotoxin (In conclusion, human hydrosalpinx fluid has a negative effect on the fertilization rate ofthe exposed mouse oocytes. However, this effect was found only in undiluted concentrationand does not affect the subsequent embryonic development and blastocyst cell number.


Control of tumour development and growth by the immune system critically defines patient fate and survival. What regulates the escape of colorectal tumours from destruction by the immune system remains currently unclear. Here, we investigated the role of intestinal synthesis of glucocorticoids in the tumour development during an inflammation‐induced mouse model of colorectal cancer. We demonstrate that the local synthesis of immunoregulatory glucocorticoids has dual roles in the regulation of intestinal inflammation and tumour development. In the inflammation phase, LRH‐1/Nr5A2‐regulated and Cyp11b1‐mediated intestinal glucocorticoid synthesis prevents tumour development and growth. In established tumours, however, tumour‐autonomous Cyp11b1‐mediated glucocorticoid synthesis suppresses anti‐tumour immune responses and promotes immune escape. Transplantation of glucocorticoid synthesis‐proficient colorectal tumour organoids into immunocompetent recipient mice resulted in rapid tumour growth, whereas transplantation of Cyp11b1‐deleted and glucocorticoid synthesis‐deficient tumour organoids was characterized by reduced tumour growth and increased immune cell infiltration. In human colorectal tumours, high expression of steroidogenic enzymes correlated with the expression of other immune checkpoints and suppressive cytokines, and negatively correlated with overall patients' survival. Thus, LRH‐1‐regulated tumour‐specific glucocorticoid synthesis contributes to tumour immune escape and represents a novel potential therapeutic target. Extra‐adrenal glucocorticoid synthesis in the intestinal epithelium controls intestinal homeostasis, inflammation and colitis‐associated tumour development in a Liver Receptor Homolog‐1 (LRH‐1/NR5A2)‐regulated manner. LRH‐1 drives proliferation and tumour‐autonomous glucocorticoid synthesis, which suppresses anti‐tumour immune responses and promotes immune escape. AOMazoxymethaneCcnd1cyclin D1Ccne1cyclin E1CTLA‐4cytotoxic T lymphocyte‐associated antigen 4CYPcytochrome p450CYP11B111β‐hydroxylaseDSSdextran sodium sulphateILinterleukinLRH‐1liver receptor homolog‐1 (NR5A2)MHCmajor histocompatibility complexNKnatural killerPD1programmed death 1SF‐1steroidogenic factor‐1 (NR5A1)SHPsmall heterodimer partner (NR0B2)TGFβtransforming growth factor βTNFtumour necrosis factorregTregulatory T cells1+ T cells has been associated with increased patient survival [+ regulatory T cells (Treg), which may limit protective immune surveillance [The control of tumour development and growth by the immune system has been first suggested more than 50 years ago ; howeversurvival , 3. Thissurvival ). The tueillance ).These findings suggest that the ability of tumours to condition their microenvironment may largely determine whether or not the immune system is able to control tumour growth and metastases. Thus, tumour‐derived factors, which limit the infiltration, activation or even survival of tumour‐specific T lymphocytes, likely substantially support tumour growth and immune evasion. The “tool kit” of cancer cells in this regard is rather extensive. Tumour cells downregulate major histocompatibility complex (MHC) class I to prevent recognition by cytotoxic T cells , releaseYet, not all patients respond to immune checkpoint inhibitors, i.e., anti‐PD‐L1 and anti‐CTLA‐4 antibodies. This suggests that different immunoregulatory mechanisms exist, which prevent the destruction of tumours by the immune system. Glucocorticoids are steroid hormones with potent immunosuppressive activities. Since their discoveries in the 1950s, they could not be missed in the treatment of numerous devastating inflammatory and autoimmune diseases. Endogenous glucocorticoids are primarily produced in the adrenal glands in response to emotional, physical and inflammatory stress. In the last two decades, various alternative sources of glucocorticoids, such as thymus, skin, lung, vasculature and intestine, have been discovered Recently, we have shown that human colorectal tumours have maintained the capability to produce immunoregulatory glucocorticoids in an LRH‐1‐dependent manner . This su22.1The cell line HEK293T had been originally obtained from Inder Verma . The authentication of the cell line was tested by PCR confirming the presence of the SV40 large T antigen. The murine intestinal epithelial cell line mICcl2 was obtained from J.‐P. Kraehenbuehl (Lausanne). The authentication of the cell line was confirmed by flow cytometry confirming E‐cadherin expression. All experiments were conducted with mycoplasma‐free cell lines.2.2ad libitum. All mice were bred under the C57BL/6 genetic background. SHP‐deficient mice (SHP−/−) [Rag1−/−) [fl/fl, LRH‐1IEC KO) were generated by crossing LRH‐1fl/fl mice with villin‐Cre transgenic mice [fl/fl) were used as controls. LRH‐1IEC KO and SHP‐deficient mice were kindly provided by K. Schoonjans . For all experiments, sibling littermates were used as controls, with the exception of SHP−/− mice, for which wild‐type C57Bl/6 mice, housed in the same room, were used.All animal experiments were approved by the local ethics committee  and conducted according to ARRIVE standards. Age‐ and sex‐matched 7‐ to 11‐week‐old mice were used for the experiments. Mice were housed in the animal facility of the University of Konstanz under specific pathogen‐free conditions with a standard diet and water (SHP−/−)  and RAG1ag1−/−)  had been2.3Cyp11b1 gene with loxP sites [fl/fl mouse line was crossed with villin‐Cre transgenic mice to generate the intestine‐specific Cyp11b1‐deficient mouse line (Cyp11b1IEC KO). The expression of Cre and the successful deletion of floxed exons 3–5 in the small and large intestines was confirmed by PCR  . Five days later, 2.2% (w/v) dextran sulphate sodium (DSS)  was given via the drinking water for five consecutive days, followed by 16 days of normal drinking water. This DSS cycle was repeated 2, resp. 3 times. Body weights were monitored. At time points, indicated mice were sacrificed, and colons were removed for macroscopic examination of tumour numbers and size, measurement of colon length, total RNA isolation and histological or immunohistochemical analysis. Tumour diameter was measured with a sliding calliper and the volume was calculated using the following formula: V = 4/3πr3 .The AOM/DSS colon carcinogenesis model was performed as described previously . Briefly2.6−/− mice were treated with 2.2% (w/v) DSS in the drinking water for 5 days followed by normal drinking water. Control mice received normal drinking water. Body weights were recorded. Mice were sacrificed on days 5, 7 and 10 after DSS treatment. Colons were longitudinally divided for RNA extraction, histological analysis (“Swiss rolls”) and ex vivo culture for measurement of colonic glucocorticoid synthesis [Female wild‐type and SHPynthesis .2.7Colonic mouse tissues were fixed in 10% formalin and embedded in paraffin. Paraffin‐embedded “Swiss roll” sections were stained with H&E for histopathological evaluation and scored microscopically for colitis using a scoring method described previously , ranging2.8Analysis of colonic glucocorticoid synthesis was done as described previously , 18, 32 2.9−/−, LRH‐1fl/fl, LRH‐1IEC KO, Cyp11b1fl/fl and Cyp11b1IEC KO mice were generated as described previously [Tumour organoids from wild‐type, SHPeviously , 34 witheviously . The culFor the analysis of tumour organoid growth, equal numbers of cells were seeded into a 96‐well plate in a drop of 8 μL BME diluted 3 : 4 with the medium. Tumour organoids were grown for 4 days, subsequently stained with Hoechst 33342, and the DNA content was analysed on a plate reader Infinite® 200 PRO  as well as by fluorescent microscopy  [2.10g, 1 h, 4 °C). Virus particles were resuspended in Advanced DMEM/F12 supplemented with 10 mm HEPES, 1× Glutamax, 1× penicillin/streptomycin and stored at −80 °C. Lentiviral transduction of organoids was performed as described [g, 1 h, 32 °C) with 8 μg·mL−1 polybrene. After incubation for 3 h, cells were seeded in BME and selection was started after 48 h with 1 μg·μL−1 puromycin.Production of lentiviral vectors was performed as described . In brieescribed . Briefly2.115 cells were resuspended in organoid basal medium, mixed with 50% BME to a final volume of 200 μL and injected subcutaneously into the flanks of C57BL/6 wild‐type mice. In some experiments, also lymphopenic Rag1−/− recipient mice were used. Tumour sizes were measured using a sliding calliper and tumour volume was calculated as 0.523 × length × width × width. At day 12 or 24 after subcutaneous transplantation, mice were euthanized, subcutaneous tumours were collected, individual tumour weight and volume were measured and tumours were used for histological and flow cytometry analysis. In some experiments, wild‐type recipient mice were treated with 0.28 mg·mL−1 dexamethasone in the drinking water ad libitum as described previously [Subcutaneous transplantation of tumour organoids was performed as described previously  with mineviously .2.12−1 collagenase IV (Sigma‐Aldrich), 40 μg·mL−1 DNase I  and 3 mm CaCl2. Digestion was stopped using 30 μL 0.5 m EDTA and cells were filtered through a 40 μm cell strainer to obtain single‐cell suspension. Dead cell staining was performed before blocking and staining steps using the Fixable Viability Dye eFluor 455UV . Cells were blocked afterwards with 3% bovine serum albumin (BSA) and anti‐CD16/32 TruStain FcX™ PLUS antibody , and stained using the following surface marker detecting, fluorochrome‐conjugated antibodies purchased from BioLegend: anti‐Ly‐6G‐BV605 (127639), anti‐NK‐1.1‐BV711 (108745), anti‐Ly‐6C‐BV785 (128041), anti‐CD26‐FITC (137805), anti‐XRC1‐PE (148204), anti‐CD64‐AF‐647 (139321), anti‐CD45‐AF‐700 (103128), anti‐CD11b‐PE‐Cy7 (101216); from eBioscience: anti‐CD11c‐PE‐eFluor 610 (61‐0114‐80), anti‐MHCII‐APC‐eFluor 780 (47‐5321‐80), anti‐CD3‐eFluor 506 (69‐0032‐80), anti‐CD19‐eFluor 506 (69‐0193‐80) and anti‐Siglec‐F‐BV421 . Flow Cytometry experiments were performed with LSRFortessa (BD Biosciences) and data were preprocessed using flowjo software .Tumours were isolated 9 or 12 days after s.c. transplantation of tumour organoids, and digested in DMEM medium containing 10% FCS, 0.75 mg·mL2.13flowjo software (BD Life Sciences). Exported FCS files were uploaded into the rstudio environment  and data transformation was performed by applying the inverse hyperbolic arcsinh function with individually set cofactors. Data were further transformed with the matrixstats r package to normalize all marker expressions between values of 0 and 1 with low (1%) and high (99%) percentiles as a limit. Each FCS sample was downsampled to 10 000 cells and combined for the Uniform Manifold Approximation and Projection (UMAP) projection using the umap r package [consensusclusterplus package with k = 27 [High‐dimensional flow cytometry analysis of tumour‐infiltrating immune cells was performed according to the workflow as previously described , 40. Com package . FlowSOMh k = 27 . Cluster2.14® Green PCR Master Mix on a One‐Step‐Plus real‐time PCR machine  and normalized to β‐actin. Primers used are listed in Table Total RNA from murine tissues and tumours was isolated using peqGOLD TriFast™  or RNA isolation kits  according to the manufacturer's instructions, and RNA was reverse transcribed. cDNA was analysed for gene expression by real‐time PCR using SYBR2.15Cyp11a1 and Cyp11b1 [The murine intestinal epithelial cell line mICcl2 was transfected with LRH‐1 expression vector, an LRH‐1 luciferase reporter (mLRH‐1 5xRE) and increasing concentrations of SHP expression vector . LRH‐1 t Cyp11b1 .2.16m Hepes, 10 mm KCl, pH 7.9, 1 mm DTT, universal protease inhibitor (Roche), 1% NP‐40) for 10 min at RT. Nuclei were pelleted at 20 000 g for 4 min at 4 °C, and lysed in buffer B  at 4 °C for 30 min. After centrifugation at 1700 g, nucleoplasm was isolated and incubated with Ni2+‐sepharose beads  for 30 min at RT. Subsequently, beads were washed 10 times in PBS, 1% Tween, 10 mm Imidazole, pH 8.0. Protein complexes were eluted in 1× PBS, 500 mm Imidazole, pH 8.0 and analysed by western blotting using anti‐Myc‐tag and GFP‐tag antibodies.HEK293T cells were co‐transfected with Myc/His‐tagged β‐galactosidase or Myc/His‐tagged SHP, and GFP‐tagged LRH‐1 expression plasmid using the calcium phosphate method. Cells were then lysed in buffer A . We next stratified the patients into two groups according to their TG expression . The Kaplan–Meier estimator provided in the R language package was used for comparing the overall survival distributions between the high versus low groups. The TCGA data and R script used to perform the Kaplan–Meier analysis are available within the Clinical data as well as gene expression (FPKM – Fragments Per Kilobase of transcript per Million) for target genes (TG) of 379 Colon Adenocarcinoma (COAD) patients with any stage ) of disease were downloaded from TCGA Data portal . Statistical tests used were unpaired Student's t‐test when comparing two groups and two‐way ANOVA followed by Tukey's multiple comparison test when comparing multiple groups. The Kaplan–Meier method was used to estimate survival distribution between the groups and log‐rank tests were applied to compare survival rates.The statistical analysis of mouse experiments was performed using For the analysis of human gene expression data, the Kaplan–Meier estimator provided in the R‐language package was used for comparing the overall survival distributions between the high versus low groups. Lastly, the comparison of immune gene expression between the high versus low groups was performed using the non‐parametric Wilcoxon's signed rank test.P‐value < 0.05 was set as the level of significance.A 33.1Lrh‐1 (LRH‐1IEC KO), suggesting increased intestinal inflammation after DSS treatment , in the intestinal tissue and the intestinal crypts ‐deficient mice  expression was only slightly increased  [We next assessed whether genetic deletion of SHP would unleash the oncogenic activity of LRH‐1 resulting in increased AOM/DSS‐induced intestinal tumours. We first confirmed that SHP expression was undetectable in the intestine of SHP   was up t3.4Cyp11a1 and Cyp11b1   Fig. . As expe1b1 Fig.  and immu1b1 Fig. , while t1b1 Fig. . These dIEC KO mice showed significantly increase body weight loss in the first two cycles of DSS, suggesting increased intestinal inflammation in the absence of immunoregulatory glucocorticoids, Cyp11b1IEC KO mice recovered faster after the third cycle of DSS  Fig. . Althougtro Fig. . When tutro Fig. . Interestro Fig. . As expetro Fig. .3.7IEC KO tumoroid transplants , but increased expression of the pro‐inflammatory cytokine IL‐6 in Cyp11b1‐deficient transplanted tumoroids. While IL‐10 expression was not significantly different, there was a tendency towards lower expression in Cyp11b1‐deficient tumours (P = 0.0837)  Fig. . These cent Fig. . To furtent Fig. . These aent Fig. .3.8CYP11A1, HSD11B1, HSD11B2, NR5A2 (LRH‐1) and NR0B2 (SHP) in tumours from 379 colorectal cancer patients, and correlated their overall survival to the expression of these steroidogenic factors. CYP11A1 (p450 side chain‐cleaving enzyme) is the rate‐limiting and thus essential enzyme in the de novo synthesis of glucocorticoids from cholesterol, whereas HSD11B2 (hydroxysteroid dehydrogenase 11B2) deactivates cortisol to cortisone, and HSD11B1 reactivates cortisone to cortisol and thus contributes to the regulation of local glucocorticoid levels , CD274 (PD‐L1) and CD152 (CTLA‐4), and the M2‐like macrophage markers LAYN, CCR4 and ARG1 [NR0B2 expression correlated with a significantly lower expression of these immune markers . Wh0B2 Fig. , but not1B2 Fig. . Interesand ARG1 . In conters Fig. . These f4It is currently well accepted that the immune system critically regulates tumour development. A limiting aspect in tumour immune surveillance, however, are all the factors that control self‐tolerance and thus limit the activation of tumour‐specific immune cells. Currently best studied are the immune checkpoint molecules PD‐1/PD‐L1 and CLTA‐4. The recognition of the importance of these immune checkpoints for controlling the immune system in general, and anti‐tumour responses in particular, has led to the development of neutralizing antibodies against these checkpoint molecules and their successful use as therapeutic agents. The results in the treatment of some but not all tumours are astonishing, in particular in combination with conventional chemotherapy. Yet, the unsuccessful treatment of many patients clearly indicates that other potential immune checkpoints may exist.in vitro deletion of the Cyp11b1 gene in tumour organoids by lentiviral Cre transduction resulted in similar reduced tumour growth upon s.c. transplantation. To what extent this immune surveillance is mediated by tumour‐specific T lymphocytes, innate immune cells, such as NK cells and neutrophils, or a combination thereof [Here, we provide evidence that tumour‐derived glucocorticoids may indeed represent such a novel “checkpoint”, and that inhibition of tumour glucocorticoid synthesis could develop as an effective therapeutic approach. Although we transplanted syngeneic tumours into syngeneic recipient mice, in the absence of dominant tumour model antigens, our data point out that inhibition of glucocorticoid synthesis in tumours alone is sufficient to promote a protective immune response, causing a strong reduction in tumour growth. Importantly, differences in genetic mutations of the transplanted tumours could be excluded as  thereof , 49, 50,de novo synthesis of glucocorticoids from cholesterol, and HSD11B1, initiating reactivation of glucocorticoids from inactive precursors , siges Fig. . A similges Fig. . In suppCyp11b1, abrogating intestinal glucocorticoid synthesis, resulted in increased inflammation and increased tumour development during the early phases of chronic colitis  deletion of SHP was able to promote the oncogenic activity of LRH‐1, resulting in increased growth of transplanted tumoroids (Fig. A major aim of this study has been to investigate the relative contribution of LRH‐1‐regulated glucocorticoid synthesis in the inflammation‐induced development of colorectal tumours. Our results point out that intestinal glucocorticoid synthesis has a prominent dual role in this process. At the early phase, LRH‐1‐regulated intestinal glucocorticoid synthesis appears to suppress intestinal inflammation and thus associated tumour development. Along these lines, we have seen that intestine‐specific deletion of tis Fig. . In contids Fig. . While tids Fig. , indicatids Fig. .CYP11A1 appears to be frequently downregulated [CYP11A1 expression results in increased tumour growth and reduced patient survival. Interestingly, certain CYP11A1 polymorphisms have also been associated with an increased risk to develop breast cancer. In addition, CYP11B1, encoding 11β‐hydroxylase, has been reported to be frequently mutated in different tumours, including colorectal tumours [CYP11A1 expression (Fig. CYP11B1, as expression was not reported for all tumour tissues summarized in the TCGA database. One of the underlying reasons may well be that basal expression levels of CYP11B1 may be lower than those of CYP11A1. This notion is in line with our previous observation in the murine intestine, where Cyp11b1 was barely detectable in control mice, but strongly upregulated upon immune cell activation or by inflammation (Fig. Cyp11b1 deletion abrogated it (Fig. While the oncogenic role of LRH‐1 has been well documented in a variety of tumours, the role of steroidogenic enzymes in the development of tumours, and colorectal tumours in particular, is less clear. In colon cancer, egulated . Nonethe tumours . Althougion Fig. , we haveion Fig.  [18, 32, it Fig. .Based on the here discussed role of tumour‐derived glucocorticoids in immune escape, LRH‐1‐regulated steroid synthesis in tumour cells may represent an attractive new therapeutic approach. However, a general suppression of glucocorticoid synthesis will likely result in severe side effects, given the widespread roles of glucocorticoids in numerous physiological processes. Thus, a detailed understanding of the tumour‐specific processes controlling glucocorticoid synthesis in colorectal cancer may be instrumental to develop tumour‐specific therapies. In this regard, LRH‐1 seems to be the most promising target.5We have previously shown that human colorectal tumour cells are capable of producing immunoregulatory glucocorticoids in an LRH‐1‐regulated manner. In this study, we now specifically addressed how tumour‐derived glucocorticoids regulate anti‐tumour immune responses and tumour growth in a murine model of inflammation‐driven colorectal tumour development. Our results show that tumours lacking LRH‐1 expression or its transcriptional target Cyp11b1 fail to produce tumour‐autonomous glucocorticoids, which results in better control of the tumour growth by tumour‐infiltrating immune cells. Thus, tumour‐derived glucocorticoids appear to be an important immune checkpoint. Comparative gene expression and survival data analysis indicate that similar immune escape mechanisms exist also in human colorectal cancer patients.The authors declare no conflict of interest.IEC KO mouse model; JK helped with the AOM/DSS tumour model; KB, CM and HFF helped with the tumoroid system; APP contributed to the figure generation; ITS and KJG analysed the gene expression databases; EK advised on clinical samples; TB supervised the study and wrote the manuscript.AA planned and conducted most experiments, and generated the figures; CR, EB, CU, MN, TSP, LD, VMM, FK and FN conducted some experiments; FK and RS characterized the Cyp11b1Fig. S1. Generation and characterization of intestine‐specific Cyp11b1‐deficient mice.Fig. S2.In vitro growth of tumour organoids.Fig. S3. Effect of dexamethasone treatment or Cyp11b1 deletion on in vivo growth of s.c. transplanted tumours.Fig. S4. Characterization of tumour immune infiltrates.Fig. S5. Effect of NR5A2 and HSD11B2 expression on colorectal patient survival and immune marker expression.Click here for additional data file.Table S1. List of PCR and qRT‐PCR primers.Click here for additional data file.


Dishonesty has received increased attention from many professionals in recent years for its relevance in many social areas such as finance and psychology, among others. Understanding the mechanisms underlying dishonesty and the channels in which dishonesty operates could enable the detection and even prevention of dishonest behavior. However, the study of dishonesty is a challenging endeavor; dishonesty is a complex behavior because it imposes a psychological and cognitive burden. The study of this burden has fostered a new research trend that focuses on cognition’s role in dishonesty. This paper reviews the theoretical aspects of how such cognitive processes modulate dishonest behavior. We will pay special attention to executive functions such as inhibitory processes, working memory, or set-shifting that may modulate the decision to be (dis)honest. We also account for some frameworks in cognitive and social psychology that may help understand dishonesty, such as the Theory of Mind, the role of creative processes, and discourse analyses within language studies. Finally, we will discuss some specific cognitive-based models that integrate cognitive mechanisms to explain dishonesty. We show that cognition and dishonest behavior are firmly related and that there are several important milestones to reach in the future to advance the understanding of dishonesty in our society. Dishonesty has been extensively studied over the past decade, as it reflects a significant interest in human behavior and has a considerable impact on professional structures such as companies and organizations . Many stWe need to suppress the truth when we lie”, or “We got to monitor whether the receptor of our scheme is suspecting something”), elements and terms related to the cognitive literature, such as suppression or monitoring, can be easily identified. Such decompositions have led to a new body of research that assumes that dishonesty has a cognitive disadvantage/burden over honesty. Several works have emerged to test for the extra resources demanded by dishonest behavior [By breaking down the concept of dishonesty , among others. Dishonesty is a complex phenomenon, as well. Some of the most frequent forms of dishonesty are cheating, lying, fabrication, sabotage, impersonation, etc. Concepts such as deception, which includes omitting information, misreporting, or misleading messages, among other similar behaviors, are also part of what we understand as dishonest behavior while not strictly being considered as lies. Recently, Srour and Py  proposedSecond, recent research has shown that dishonest behavior should not be dichotomized as honest/dishonest. Dishonesty seems to be within a continuum or “grey scale” of different levels of dishonest behaviors ,9. TradiHence, we have structured the present review while trying to answer the following questions: Which are the main cognitive processes related to dishonesty? How does social cognition modulate dishonest behavior? How are other cognitive processes related to deception? Which cognitive models can help us to understand dishonesty?According to the literature, executive functions (EF) are the most significant cognitive processes for understanding dishonest behavior. Most research studying cognition and dishonesty has focused on EF. Thus, we will first examine the most important studies on EF and dishonesty, specifically on inhibition, working memory, and task switching—three of the most relevant cognitive processes defining EF. Although we will discuss at some points how these cognitive processes are implemented in our neural structures and circuits, our focus will be on the cognitive processes themselves, paying less attention to structural brain implementation or connectivity. There are other works for readers interested in an extensive review of the neuroscience of dishonesty ,12. WhilDishonesty is essentially a social behavior. It depends on others’ perceptions and how they interact with our world understanding. Therefore, a critical question is how dishonesty may interact with social cognition. The cognitive Theory of Mind (ToM) is the ability to understand others’ beliefs. Thus, it seems to be essential to understanding dishonesty. Furthermore, although discussing how personality traits can explain dishonest behavior is beyond the scope of this review (and would warrant a separate review in itself), we will talk about some related factors, such as creativity. We will review some interesting work on dishonesty and creativity, as well as other work reviewing the role of language in dishonest behavior.Finally, we will review several cognitive models that can provide insights into understanding dishonesty and how the different cognitive processes previously described are interconnected. To summarize, as the field of dishonesty continues to evolve, this review aimed to explore the literature and examine some of the key insights and major breakthroughs made thus far in understanding the relationship between cognition and dishonesty. We will finish the review by discussing limitations and the new challenges to come.Dishonesty is exhausting. Those who have tried to uphold a lie for a long time likely understand how exhausting and cognitively demanding it can be. Studying the relationship between cognition and dishonesty has become critical to understanding this demanding and complex behavior. As previously mentioned, one of the earliest works to suggest this relationship was Zuckerman et al. , which eVrij  notes thIn the study of dishonesty and working memory (WM), most researchers in the field have employed dual-task paradigms to investigate how dishonesty is affected by increased cognitive loads. The underlying assumption is that maintaining a lie or engaging in deceptive behavior requires additional cognitive effort, and this should be reflected in changes in behavior and performance under increased WM load conditions. For example, Vrij et al.  found thTo address this question, Van’t Veer et al.  used theIn a recent study, Speer et al.  showed tFurther evidence on the impact of working memory (WM) on dishonesty is derived from studies on child development. These studies are particularly intriguing because executive functions are not fully developed during childhood. In fact, depending on their age, differences in WM capacity can play a crucial role . AllowayTherefore, WM load appears to influence or moderate dishonesty. New theoretical models including WM as an explanatory factor suggest that WM’s role in dishonesty can be twofold: it can come from the need to retrieve information required for the task, but also to update new information, items, and targets as the behavior evolves . In thisDeliberate actions require cognitive control to overcome instinctive responses . This prHowever, other researchers using different types of inhibitory tasks have not found significant differences between lying and telling the truth. Fenn et al.  reportedIn their study, they first measured inhibition with a stop-signal task, and in another session, participants performed a Sheffield Lie Test. However, they did not replicate the previous findings and found no differences between conditions of lying and telling the truth, nor any relation with the function of the stop-signal task score among individuals. Aïte et al.  attempteHowever, more research is needed to understand the fundamental mechanisms of inhibitory processes in dishonesty. The study of task switching (set-shifting), which involves changing the mental set between tasks, could provide more clues about the functioning of inhibitory processes in dishonesty.Dishonesty is not always a clear-cut binary or radical action; there are several levels at which dishonest behavior can occur. For instance, one can choose to not divulge the complete truth in a given situation, resulting in the omission of relevant information, but not as a direct lie. Furthermore, not all acts of dishonesty are completely dishonest. In Pascual-Ezama et al.’s study , particiIn a meta-analysis, Christ et al.  demonstrSimilarly, Foerster et al.  studied These studies provide evidence for the cognitive burden of dishonesty and highlight the importance of studying executive functions in dishonest settings to understand and potentially prevent cheating, lying, and other forms of dishonesty. However, as dishonesty is a complex behavior, it cannot be fully understood by a single approach. Recent models of dishonesty have considered executive functions and other important processes, such as social cognition and creativity, to explain the mechanisms of dishonesty. The social nature and transgressive character of unethical behavior have led to linking dishonesty to paradigms like the Theory of Mind, with executive functions playing a role in these theories ,41. In tDishonesty has a social dimension, which, with its cognitive nature, makes social cognition processes crucial for understanding it. Dishonest behavior often occurs in a social context and thus requires an understanding of both the context and the intentions of the individuals involved . Models ToM is the ability to reason about others’ mental states and beliefs . It usuaThere are also studies of the ToM and dishonesty with adults. Apperly et al.  studied From another perspective, Johnson et al.  carried Thus, empirical evidence supports ToM as being necessary to perform dishonest behavior. However, some questions remain open to understanding how social cognition, together with other cognitive processes, works in dishonesty.The literature reviewed so far has emphasized the significance of executive functions in conjunction with social development to explain dishonest behavior. However, there may be other cognitive processes to consider to better comprehend dishonest behavior, such as creativity and language. The well-known Zuckerman’s model analyzes dishonest discourses and compares the paraverbal elements in language between honest and dishonest behaviors to detect dishonesty. The literature has identified several significant differences in the production of dishonest speech, including shorter length, more pauses and hesitations, more errors, and vaguer details ,60 . Language is also explicit in the first assumption, as it particularly applies to verbal and non-verbal behavior. Emotions considered in the third assumption can perfectly imply social aspects, as they involve others to be guilty with, for instance, and probably ways to control self-representations and self-awareness. The increase in arousal postulated in the second assumption could also be related to attentional and inhibitory aspects, as they imply focusing on something to avoid something else. Thus, in general, the model proposes an interrelation of different aspects explaining dishonesty that involves several of the cognitive processes described in the previous lines of this review.The Interpersonal Deception Theory (IDT)  posits tSimilarly, the Truth-Default Theory (TDT)  uses conAnother relevant model in the field is the Activation–Decision–Construction–Action Theory (ADCAT) proposed by Walczyk et al. . This moThe Information Manipulation Theory 2 (IMT2)  is a varIn contrast to previous models that focused on contextual and discourse approaches, Sporer  proposedMoreover, the “schemata” concept raised in several models is critical in Sporer’s model. According to the schemata approach , sequencSimilarly, Lane and Wegner  proposedThis review of evidence on dishonesty that has been collected over the past few decades shows the significance of cognitive processes explaining and understanding dishonest behavior. A key assumption underlying the link between dishonesty and cognition is the effort required to sustain dishonest behavior in our cognitive system. This assumption is backed by empirical data on the need to suppress automatic honest responses, the constant monitoring of one’s behavior and discourse while being dishonest, and the evaluation of context and inferences about other people’s involvement. All these assumptions support a theoretical framework based on human capacity limitations, where dishonesty is a complex behavior that demands constant activation and deactivation of information and requires sufficient resource availability to occur .However, as we have seen, there are some contradictory results among studies. As in many other research fields, these differences could result from the diversity of manipulations and paradigms among studies. More research is needed to disentangle the puzzle of how cognition modulates dishonesty. However, despite the variations among models and studies, there are some common agreements. Most suggest that dishonesty is a process that can be broken down into stages that involve monitoring (working memory), inhibition, and set-shifting of information. The specifics of how this occurs vary between models and studies and the empirical data supporting each of them. For instance, certain models postulate that specific general cognitive processes, such as inhibition, might affect behavior differently depending on the dishonest task manipulation or contextual variables ,31. InhiIt is crucial to note that incorporating new experimental tasks may enhance our understanding of how cognition operates in dishonest behavior. Pascual-Ezama et al.  developeHowever, although the present work has a significant contribution and strength in its multidisciplinary approach to addressing cognition involvement in dishonesty, that also brings some limitations. Given dishonest behavior’s complex and extensive nature, we aimed to review most breakthroughs using diverse experimental paradigms. The paradigms reviewed here are summarized in In conclusion, the growing research interest in the cognitive mechanisms underlying dishonest behavior has resulted in valuable insights into how dishonesty operates. These advances in scientific understanding may even pave the way for detecting and preventing dishonest behavior. A key aim of studying dishonesty is to find ways to mitigate its impact on professional structures, companies, organizations, politics, and daily human interactions. The investigation of cognition in dishonesty can help us achieve this goal.