Goal pursuit shapes people’ everyday experiences and is deeply embedded within close relationships. Several studies have shown that goal support from romantic partners facilitates goal progress, and individual goal progress contributes to wellbeing. However, few pieces of research have examined the whole process, how efficient goal coordination in a romantic relationship contributes to life satisfaction through goal progress. In these studies, short time frames were used and only one aspect of goal coordination was examined. To generate more complex, long-term understanding we collected data from 148 married or cohabitating Hungarian heterosexual couples  in a two-wave longitudinal study with a year-long time window. Both partners individually completed an adapted version of the Personal Project Assessment and evaluated four chosen projects associated with project coordination  at baseline, and project attainment  in the follow up. Life satisfaction was assessed during both waves. Results from the actor–partner interdependence mediation modeling revealed complete mediation, where project coordination increased project attainment one year later, and consequently associated with higher life satisfaction for both partners. The direct effect between project coordination and life satisfaction remained nonsignificant. This association indicates that for long-term life satisfaction, it is crucial to experience better goal outcomes as the result of the couple’s collaborative effort. Goal pursuit shapes how people experience their lives. They may strive to be promoted, organize a vacation, or lose some weight, while also trying to be good partners to their loved ones. Progressing with or attaining personal goals increases individual well-being , conceptualizes people in close relationships as one self-regulating unit based on the strong interdependence of members’ goal setting, pursuit, and outcomes  was identified as positively related to goal progress over time, but autonomy support did not predict change in subjective wellbeing, even though significant associations were found in the other studies reported in the article with non-romantic dyads  increased goal commitment over time  have been cohabitating for at least one year, 2) be between 25 and 65 years old, 3) have no previous history of psychiatric disorders in the past five years, and that 4) at least one spouse has active working status. Two couples were excluded from the analysis for not meeting at least one of the criteria. For our final sample, 148 couples were available for the second wave of the study, while 65 couples refused or were unavailable.The mean age was 39.72 years for men (SD = 10.40) and 38.57 years for women (SD = 10.00). The average relationship length was 17.10 years (SD = 9.99 years). Regarding relationship status, 64 couples (43.24%) were cohabiting without marriage, and 84 were married (56.75%). More than two-thirds (70.27%) of men and 68.92% of women reported to having at least one biological child . 18.07% of participants had earned college diplomas (16.89% of men and 19.26% of women), 63.85% had earned a high-school diploma (66.22% of men and 61.49% of women) and 16.89% had completed only primary education (20.95% of men and 12.84% of women).Prior to the data assessment we acquired ethical approval from Semmelweis IRB (SE TUKEB). Potential participants were contacted at their homes. After both partners provided their informed consent, which included their inclusion in a one-year follow up, the interviewer administered the questionnaire pack and explicitly instructed the spouses to complete the assessment procedure separately. Then the interviewer left participants to fill out the survey by themselves, and recollected completed questionnaire pack at a later date agreed by phone. All data collected were handled confidentially. All partners that participated in the study voluntarily and received a book voucher for their contribution  in each wave of the study. Data sampling occurred from 2013 to 2014, which makes the sample unaffected by COVID-19 pandemic.t(182) = -2.236, p = 0.027).The same procedure was repeated one year later. To examine potential drop-out bias, we compared the sample of retained to lost couples. There were no differences in the goal-related or well-being variables under analysis at baseline. Regarding the demographic variables, we detected no difference in age, relationship status, numbers of children, or level of education, but couples who withdrew from the study had relationships shorter by three years on average , using three items – based upon communication (‘How frequently do you communicate with your partner about this project?’), cooperation (‘How frequently do you cooperate with your partner on this project?’) and the partner’s perceived emotional support . The endpoints of the Likert scales were noted as 1 = ‘very rarely’ and 7 = ‘very often’. The internal consistency of this scale was acceptable (α = 0.754 for men and 0.781 for women partners).During the second wave (Time 2), participants were presented the four projects they selected at the first wave and they were instructed to evaluate, how much they attained from each project using the following items: ‘How far have you progressed with this project?,’ How successful were you in the accomplishment of this project?’  and ‘Taken together, how satisfied are you with the way things have gone with this project over the past year?’ . The internal consistency of this scale was excellent for both genders (α = 0.972 for men and 0.965 for women partners).p < 0.001) and medium correlation for women partners .The Satisfaction With Life Scale  , which Because of the high complexity of the model, a sensitivity analysis was conducted for the final APIMeM, where we used the sum scores instead of latent factors for SWLS’, based on its high internal consistency  was used for all analyses. We evaluated model fit for baseline models and hypothesis testing based on CFI, RMSEA, and the Standardized Root-Mean-square Residual (SRMR). The model was considered to fit the data well with a CFI value of 0.90 or higher, an RMSEA value of 0.06 or lower, and a SRMR value of 0.08 or lower Kline, . All anap < 0.001), while project attainment items were positively related as well . Following the hypothesized pattern, items related to project attainment showed significant positive correlation with SWLS at the second wave . Women’s SWLS was significantly associated with support  and items related to project attainment . Correlation between items of project coordination and project attainment was non-significant in most cases with the exception of men’s communication and success . In general, couples had similar expressions in project-related experiences  and life satisfaction  similarly to their partners.Descriptive statistics and correlations of the Personal Project Assessment variables and SWLS are shown in Table 2 (42) = 38.257, CFI = 1.000, RMSEA < 0.001 for PPA variables and χ2 (154) = 258.59, CFI = 0.956, RMSEA = 0.068 for SWLS). The results provide evidence for metric invariance across gender and timepoints for PPA variables (Δχ2 (4) = 3.507, p = 0.476, ΔCFI < -0.001, ΔRMSEA < 0.001 for the metric against the configural model; Δχ2 (5) = 529.785, p < 0.001, ΔCFI = -0.346, ΔRMSEA = 0.263 for the scalar against the metric model) and SWLS (Δχ2 (11) = 14.798, p = 0.191 ΔCFI = -0.001, ΔRMSEA = -0.001 for the metric against the configural model; Δχ2 (16) = 471.335, p < 0.001, ΔCFI = -0.191, ΔRMSEA = 0.078 for the scalar against the metric model).Table 2 (8) = 10.43, p = 0.236), and the resulting model had a good fit to the data (χ2 (450) = 638.070, CFI = 0.954, TLI = 0.949, RMSEA = 0.053, SRMSR = 0.080), which was retained as final for the interpretation.First, we constrained all direct effects across the dyads to test for gender invariance. We found no evidence for gender effects , which in turn was associated with higher life satisfaction . The direct path between project coordination and life satisfaction was not significant . As for partner effects, only the partner’s project attainment was associated with higher life satisfaction . However, it was not significant between the partner’s project coordination and project attainment  and between the partner’s project coordination and life satisfaction . All effects for life satisfaction between the first and the second timepoint were significant .As Fig. b = 0.052, p = 0.038). and the actor-partner indirect effects  were significant. All other indirect effects remained nonsignificant.The results from testing the indirect effects supported the two patterns of complete mediation  might enrich the research question and expand on the dyadic aspects of goal coordination and attainment. Lastly, not only romantic couples can be conceptualized as one self-regulating unit, but any interdependent close relationships, including coworkers, friends and family members as well. Further studies could broaden the scope and examine the connection between goal coordination, goal attainment and life satisfaction in other types of close relationships.Our results shed light on the additional benefit of accounting for the relationship context of individual goal pursuit in understanding life satisfaction. It emphasizes the importance of efficiency in goal coordination, as the effort only might be insufficient for life satisfaction. If partners feel that their goals are supported by their spouses, they might temporarily feel better, but this will preserve as long-term life satisfaction only if their goal coordination efforts are fruitful: when they can genuinely flourish together. In praxis, it might be highly beneficial to examine not only whether the couple coordinates their resources surrounding each other’s goals, but also facilitate the efficiency of their efforts.


Hand‐foot skin reaction may influence the effectiveness of patients' treatment, patient quality of life, and the economics of health care. An effective prophylactic dermatological cream for preventing sorafenib‐induced hand‐foot skin reaction (HFSR) is yet to be identified.The aim of this study is validated the prophylactic efficacy of urea‐based creams on sorafenib‐induced hand‐foot skin reaction in patients with advanced hepatocellular carcinoma.This was a randomised double‐blind experimental study. A total of 129 patients with advanced HCC were randomly assigned to three groups. The comparison group received best supportive care (BSC), group A received BSC plus a moisturising cream, and group B received BSC plus a 10% urea‐based cream. Incidence of HFSR and cutaneous wetness were assessed 3 days before starting sorafenib and each week after starting sorafenib for 8 weeks.p > .05; comparison group/B group, p > .05). Group B reported significantly better cutaneous wetness of hands in the seventh week after starting sorafenib (p < .05) and of feet during the first 6 weeks (p < .05–.001).No significant difference was observed in the incidence density of sorafenib‐induced HFSK (comparison group/A group, This study found a nut size amount of a 10% urea‐based cream applied twice a day can maintain patients' cutaneous wetness in the first 6 weeks after starting sorafenib than moisturising‐alone cream. But it cannot reduce the occurrence of HFSR. Thus, the result supports nut‐size dose of the 10% urea‐based cream three times a day may be an appropriate dose to prevent HFSR.Clinical Trail Registration Number: NCT04568330. The sample size was estimated using G. power software version 3.1, which was established using logistic regression, odds ratio: 3.8, and power: 0.80.2.2This study population comprised patients with the following characteristics: (1) HCC by proof of pathology; (2) the presence of a tumour thrombus in the main trunk of the portal vein or the first‐order branches of the portal vein with minimal or no ascetics by abdominal CT scan; (3) Child‐Pugh liver function class A; (4) plans to receive oral sorafenib 400 mg twice per day; (5) age of 20 years or above; and (6) ability to communicate in Chinese, Taiwanese, or Hakka. Patients with (1) encephalopathy, psychosis, cognition impairment, blindness, or a hearing impairment; (2) allergic history to urea; (3) ulceration, blisters, infective problems on the palms or soles; or (4) who had undergone previous surgery, systematic chemotherapy, or frequent radial ablation were excluded.2.3The comparison group who received BSC alone were (1) informed of potential presentations of HFSR, (2) asked to wear waterproof gloves before undertaking household work or working with water, (3) provided with a method of contacting health care specialists for confirmation of an early diagnosis of HFSR, and (4) asked to self‐report when they experienced symptoms of HFSR. Group A received the same interventions as the comparison group but were additionally given the moisturising cream  for use on nine occasions and were instructed how to use the cream. The instructions were to (1) use the cream twice a day from 3 days prior to starting sorafenib and each week thereafter, (2) scoop out a nut‐sized amount with a unique spoon each time, (3) gently apply the cream evenly on symmetrical palms below the wrists and symmetrical soles below the ankles each time, and (4) wear unique cotton gloves for 30 min immediately after application of the cream. Group B received interventions similar to those of group A, except that they were given a cream container with a different component . The external appearance of the containers with the two kinds of cream was the same. The creams were white and grey in colour.2.4Data on confounders and two end‐points, incidence density of HFSR and cutaneous wetness, were collected. The confounders included gender, age, number of chronic illnesses, number of metastatic regions as well as levels of white count, haemoglobin, bleeding time, liver enzymes, albumin, and electrolyte.r = .43, p < .05). An endpoint for incidence of HFSR is patients with grade 1 symptoms.The grades of HFSR were assessed using NCI‐CTCAE version 4. This is an available psychometric patient‐reported instrument developed by Dueck et al.The level of cutaneous wetness was analysed using a skin scanner , which comprised bioelectrical‐impedance analysis (BIA) equipment. BIA is a safe, noninvasive, rapid, reproducible, portable, and inexpensive method of using simple equations for safely and accurately estimating the body water with a correlation coefficient of 0.996.2.5Data analysis was conducted using SPSS software . Descriptive statistics, including means, SDs, and frequency distributions, were used to describe patient demographic data. Analysis of variance was performed to compare the confounders between groups. Cox regression was employed to compare the incidence density of HFSR and a generalised estimating equation was used to compare the percentage of cutaneous wetness between groups, factors, and multiple time points.33.1As noted previously, 129 patients were randomly assigned to three groups. Table 3.2The incidence density of HFSR is presented in Table p > .05) as well as between the comparison group and group B .A comparison of the incidence density of HFSR among the groups is presented in Table 3.3p > .05). No significant difference was found between the varying time points of comparison groups (p > .05) as well as between the comparison group and group A (p > .05). Nevertheless, a significant difference was observed between the comparison group and group B after starting sorafenib. On the hands, group B exhibited better cutaneous wetness than the comparison group in week 7 (p < .05). On the feet, group B exhibited better cutaneous wetness than the comparison group in week 1 (p < .05), week 2 (p < .05), week 4 (p < .05), week 5 (p < .05), and week 6 (p < .001).The majority of patients had dry skin on their hands or feet at different time points. The cutaneous wetness was 32.35%–33.31% . A case manager obtained informed consent from all patients prior to their participation in the study. The study protocol was explained to the patients and they were taught self‐care skills by the case manager. All were aware that their rights would be protected, that the risk of participation would be the lowest possible, and that the most suitable form of management would be arranged when they develop sorafenib‐induced HFSR.This study was approved by the Institutional Review Board of the Taipei Veterans General Hospital (2013‐13‐009B) and has been registered on website of R.‐Y.L.: Conceptualization; data curation; formal analysis; investigation; methodology. H.‐H.T.: Conceptualization; methodology; writing – review and editing. S.‐L.W.: Conceptualization; data curation; formal analysis. H.‐L.H.: Conceptualization; writing – review and editing. S.‐F.L.: Data curation; investigation.


Sebastes contains numerous species pairs that are depth‐segregated and can provide a better understanding of the mode and tempo of genomic diversification. Here, we present exome data on two species pairs of rockfishes that are depth‐segregated and have different degrees of divergence: S. chlorostictus–S. rosenblatti and S. crocotulus–S. miniatus. We were able to reliably identify “islands of divergence” in the species pair with more recent divergence (S. chlorostictus–S. rosenblatti) and discovered a number of genes associated with neurosensory function, suggesting a role for this pathway in the early speciation process. We also reconstructed demographic histories of divergence and found the best supported model was isolation followed by asymmetric secondary contact for both species pairs. These results suggest past ecological/geographic isolation followed by asymmetric secondary contact of deep to shallow species. Our results provide another example of using rockfish as a model for studying speciation and support the role of depth as an important mechanism for diversification in the marine environment.Speciation in the marine environment is challenged by the wide geographic distribution of many taxa and potential for high rates of gene flow through larval dispersal mechanisms. Depth has recently been proposed as a potential driver of ecological divergence in fishes, and yet it is unclear how adaptation along these gradients' shapes genomic divergence. The genus  Sebastes) that are depth‐segregated. We found shared islands of divergence between the two species pairs and found neurosensory genes enriched in these islands for one species pair. Demographic histories were similar for the two species pairs, suggesting similar modes of diversification.To determine the genomic responses and demographic history of speciation, we sequenced exome‐enriched sequences in species pairs of rockfish (genus  The two species pairs occur along a continuum of divergence, with S. chlorostictus–S. rosenblatti diverging approximately 0.21 Mya and S. crocotulus–S. miniatus diverging approximately 2.3 Mya , the Japanese medaka , and green‐spotted puffer . Importantly, certain genetic regions may be absent or highly divergent within the genome of this sculpin but remain conserved in other Perciformes. To account for these regions and ensure their capture, we iteratively designed capture baits from the genomes of the shorthorn sculpin Myoxocephalus scorpius (ASM90031295v1)  (7), the golden redfish S. norvegicus (ASM90030265v1) , the rougheye rockfish S. aleutianus (ASM191080v2), the European perch Perca fluviatilis (ASM90030264v1) . For each species, we included new capture baits if the targeted elements were either not identified , or had <85% identity to an existing capture bait. As a result of this iterative addition of sequence information from new species, there should be oligonucleotide capture baits of at least 85% identity to each clade included in the capture design. This multi‐species design enables efficient enrichment across distantly related perciform fishes. The final species composition of the capture baits: C. rheanus (62.0%), M scorpius (6.7%), A. fimbria (6.4%), S. norvegicus (5.9%), S. rubrivinctus (2.0%), S. aleutianus (1.8%), P. fluviatilis (5.3%), and G. aculeatus (9.8%).We used BLASTN  to identify each targeted element within multiple perciform genome assemblies. The majority of capture baits were designed against the genome of the Chabot de Rhénanie SeqCap EZ Developer (cat #06471684001) capture oligos were designed in collaboration with the Nimblegen design team to standardize oligo annealing temperature, reduce probe redundancy, and remove low complexity DNA regions. The capture design contained sequence from 492,506 regions  across all eight perciform reference genomes. Accounting for probe redundancy between the perciform reference genomes, the final capture design comprised 407,084 distinct elements, including 285,872 protein‐coding exons, 118,406 conserved non‐coding elements, 2508 UCNEs, and 298 miRNAs  to reduce the signal from false positives.To identify divergence islands, we applied an approach similar to Holliday et al.  and Rena2.6D. rerio genome from the Gene Ontology Consortium . We explored seven simple two‐population models in δaδi: no migration, symmetric migration, asymmetric migration, symmetric migration followed by isolation, asymmetric migration followed by no migration, secondary contact with symmetric migration, and secondary contact with asymmetric migration. We hypothesize that if ecological speciation has occurred in these species pairs, due to adaptation to different depths, we would expect to observe a demographic history with at least some gene flow. We utilized the δaδi optimization procedure from Portik et al. (https://github.com/dportik/dadi_pipeline). We ran four iterations of optimizations for each model with 10, 20, 30, and 40 replications, respectively. For each species pair, we compared models using AIC and Δ AIC. We did not convert parameters from the best fit model into biologically meaningful values, as our goal was simply to reconstruct a reasonable demographic scenario for each species pair.We utilized folded site frequency spectra in δaδi  and 33 windows for S. chlorostictus–S. rosenblatti . Eighteen of the 200 candidate genes in this species pair were enriched within these GO terms. Of the 18 genes: five are related to ATP binding, ATP synthase or ATPase activity , four are solute carriers , and two are integrin beta subunits (itb4r and itb4r2a). The remaining seven genes have functions related to melanophore production stimulation, germ cell migration, behavioral, ectodermal placode development, cell proliferation inhibition, and MHC class I binding activity  (Table Genes found within significant t) Table .FST windows for S.crocotulus–S. miniatus were enriched for pathways related to bicellular tight junction, positive regulation of actin filament polymerization, histone lysine methylation and SWI/SNF superfamily‐type complex (Bonferroni <0.05). Alone, 12 candidate genes from a list of 126 candidate genes were enriched for these GO terms and provided functional and cellular pathway insight. Of the 12, three are involved in chromatin remodeling , three are involved in methylation , four are related to cytoskeletal organization , one to GTP binding (cdc42ep4a), and one to intracellular membrane organization (rab13) (Table Genes found within significant tion rab1 (Table 33) Table .3.4S. rosenbaltti–S. chlorostictus and S. miniatus–S. crocotulus species pairs, respectively. For both species pairs, we found the secondary contact with asymmetric gene flow from the deep to shallow species to be the best model  that has a recent divergence (similar to S. chlorostictus–S. rosenblatti). A more direct comparison of divergence islands with our study is not entirely possible, as Behrens et al.  and driven by changes in sea level during the Pleistocene. Support for the secondary contact model found in this study is also supported by other studies of speciation history in marine organisms  is followed by a period of isolation in these species pairs. Models of ecological speciation predict that gene flow should persist upon invasion of the new ecological space. However, a recent study on depth‐segregated ecomorphs in 4.3(S. crocotulus–S. minatus), and future work in this area should focus on more recently derived species.Our work is limited in that we utilized a pool‐seq approach and only focused on enriched exome, CNE and UCE sequences. The main advantage of the pool‐seq approach is that it reduces the overall cost of sequencing , which has been observed in other marine taxa. These results build on the growing knowledge of speciation history in the genus Sebastes and suggest Sebastes will continue to be a valuable model in understanding mechanisms of speciation in temperate marine fishes.Our exome scan of two Daniel Olivares‐Zambrano: Data curation (lead); formal analysis ; investigation ; methodology ; validation ; writing – original draft ; writing – review and editing . Jacob Daane: Methodology ; writing – review and editing . John Hyde: Data curation ; resources ; writing – review and editing . Michael Sandel: Resources ; writing – review and editing . Andres Aguilar: Conceptualization ; formal analysis ; funding acquisition (lead); project administration (lead); supervision (lead); writing – original draft ; writing – review and editing .None declared.Appendix S1Click here for additional data file.


Although much has been written on the topic of social behavior, many terms referring to different aspects of social behavior have become inappropriately conflated and the specific mechanisms governing them remains unclear. It is therefore critical that we disentangle the prosocial and antisocial elements associated with different forms of social behavior to fully understand the social brain. The lateral septum (LS) mediates social behaviors, emotional processes, and stress responses necessary for individuals to navigate day-to-day social interactions. The LS is particularly important in general and selective prosocial behavior (monogamy) but its role in how these two behavioral domains intersect is unclear. Here, we investigate the effects of chemogenetic-mediated LS activation on social responses in male prairie voles when they are 1) sex-naïve and generally affiliative and 2) after they become pair-bonded and display selective aggression. Amplifying neural activity in the LS augments same-sex social approach behaviors. Despite partner preference formation remaining unaltered, LS activation in pair-bonded males leads to reduced selective aggression while increasing social affiliative behaviors. These results suggest that LS activation alters behavior within certain social contexts, by increasing sex-naïve affiliative behaviors and reducing pair bonding-induced selective aggression with same-sex conspecifics, but not altering bonding with opposite-sex individuals. Selective DREADD-mediated activation of the lateral septum reveals its importance in enabling male prairie voles to adjust selective and nonselective social behavior to account for important life-history shifts linked with reproductive success. For as long as people have studied social behavior, an underlying goal has been to understand the mechanisms that govern them. Neuroscientists with different backgrounds and expertise have attempted to define the core neural mechanisms that underlie social behavior, but defining a “social brain” is as complicated as the behaviors that are presumably under its control. This is because social behavior takes so many different forms , each of which involves many different behavioral elements and interactions. Nevertheless, determining how the brain processes and shapes behavior in response to social factors is of great importance if we are to truly understand the nature and universality of social behavior.1 provided an expanded view of the neural control and modulation of social behavior, and introduced the idea of the social decision-making network (SDMN). The SDMN is comprised of interconnected neural structures that are heavily involved in the regulation of social behavior2. Contained within the SDMN are sub-circuits that are implicated in coordinating different aspects of social behavior, including social grouping5, aggression6, parental care7, and pair bonding9. For example, work in prairie voles has defined a neural circuit that governs pair bonding, which has formed the basis for understanding the neurobiology of mammalian social affiliation and monogamy (reviewed in refs. 13). Notably, the entirety of the pair bond neural circuit is encompassed by the SDMN. In contrast to affiliation and sociability, aggression and territoriality can be considered examples of the “dark side” of social behavior. Nevertheless, aggression is a prevalent and fundamental aspect of social behavior that enables defense of resources, offspring, and mating partners. Not surprisingly, neural structures within the SDMN also play a prominent role in the regulation of aggression, and aggressive responses to intruders can be precisely modified through manipulations of sub-units of the SDMN6. The lateral septum (LS) is a central node of the SDMN and plays a prominent role in the regulation of social behavior across vertebrates1. For instance, manipulations of the LS modify social grouping preferences in finches5, aggression in mice6, kin preferences in rats14, and pair bonding in prairie voles16. Recently, Kelly et al.17 demonstrated that social approach is associated with early-life social experience and epigenetic modification of the vasopressin receptor within the LS. Ultimately, the LS appears to impact prosocial and antisocial aspects of social behavior across contexts and species.O’Connell and Hoffmann18. Prairie voles (Microtus ochrogaster) form long-term pair bonds with mates, form intense attachments with offspring, display selective aggression to defend territories and mates, and exhibit bi-parental care for their young23. Importantly, sexually inexperienced male and female prairie voles are rarely aggressive toward conspecifics and typically spend most of their time engaging in affiliative and investigative behaviors24. Mating in prairie voles facilitates the formation of a pair bond and creates a dramatic shift from engaging in general affiliation with familiar and novel conspecifics, to exhibiting selective affiliation with familiar conspecifics and aggression toward strangers. Specifically, pair bonded voles preferentially engage in partner- and offspring-directed affiliative behaviors, become profoundly territorial, and exhibit high levels of aggression toward unrelated males and females that trespass their territory. Thus, despite the common caricature that prairie voles are “highly social”, they display a wide range of prosocial and antisocial behaviors according to their physiological and environmental states. Thus, the range of context-dependent prosocial and antisocial behavior in prairie voles raises the question: what is the neural basis for this flexibility in social behaviors that enables effective reproduction?Prosocial and antisocial behaviors profoundly impact social monogamy, parental investment, and group structure26. The ability to achieve these adaptive behavioral responses to social stimuli and environmental constraints relies upon efficient transmission, appraisal, and processing of information via dynamic neural and molecular mechanisms27. Neuromodulation of the LS could serve this function. Sheehan et al.28 have proposed that the LS appears to play a regulatory function for social behavior. They argue that it does so by integrating sensory stimuli and assessing their affective relevance and valence. The LS then conveys this information to other SDMN brain regions known to promote emotional states and/or for directing motivated behaviors. Thus, the LS can orchestrate behavioral responses by appropriately adjusting behavior to meet environmental demands28.Capitalizing on resources and social interactions requires animals to appropriately adjust their behavioral output by calibrating the intensity and duration of prosocial and antisocial responses targeted at familiar or novel conspecifics29, we aim to understand the modulatory role of the LS on regulating male prairie vole social behavior in the context of mating decisions. Here, we hypothesize that the LS serves as a point of convergence for the contextual regulation of numerous aspects of social behavior central to reproduction. To this end, we assess the impact of LS chemogenetic stimulation on affiliation, aggression, and partner preference formation in male prairie voles. We evaluate the function of the LS before and after sexual experience with females to assess how LS activation impacts the transition from being generally sociable to socially selective, a shift that characterizes pair bonded prairie voles. We conclude that the LS is positioned to integrate social context-dependent information to produce flexible and appropriate behavioral responses.As an often-overlooked component of the social brain30, a second group of control adult males did not express the hM3 receptor in the LS and were injected with 3 mg/kg C21  into the LS, followed by intraperitoneal (i.p.) injections of 3 mg/kg of compound C21  = 5.35, p = 0.007; Fig. t35.3 = −2.76, p = 0.009). LS activation (hM3 + C21) did not alter the latency to social approach  = 0.97, p = 0.39; Fig. F = 0.004, p = 0.10; Fig. F = 3.19, p = 0.06; Supplementary Fig. F = 3.21, p = 0.06; Supplementary Fig. We first assessed behavioral responses to LS activation by evaluating social approach behaviors in sex naïve males on day 16 Fig. . We founF,64 = 5.3F,64 = 5.3F = 20.98, p = 2.51 × 10−5, Fig. t32.4 = 2.53, p = 0.008; sham + C21, t32.4 = 3.09, p = 0.002), as did males whose LS was activated . Moreover, partner preference indices [(time with partner − time with stranger]/time with partner + time with stranger]) for each treatment group  = 0.18, p = 0.84; Fig. F = 1.04, p = 0.37; Fig. We investigated whether LS activation would alter the ability of males to form socially selective partner preferences. Across treatments, males spent more time in contact with the partner than the stranger  = 0.16, p = 0.85; Supplementary Fig. Importantly, cage transitions were not affected by LS activation  and after (day 20) pair bond formation in the resident-intruder test  = 7.59, p = 0.0001; Fig. t35.3 = 2.03, p = 0.05; and sham + C21: t35.3 = 2.97, p = 0.005) engaged in a greater duration of attack behaviors when they were pair bonded compared to when they were sex naïve . In contrast, LS activation significantly inhibited pair bond-induced attack behaviors directed at the intruder when males were pair bonded compared to when they were sex naïve .Duration of attacks increased after forming a pair bond compared to when males were sex naïve  = 5.75, p = 0.007) and bond status  = 19.03, p = 0.0001). The interaction effect fell short of our significance threshold  = 2.92, p = 0.07). Post hoc analysis revealed that, as expected, sex naïve males did not differ in latency to attack . However, once pair bonded, hM3 + saline males significantly decreased their latencies to attack in comparison to when they were sex naïve , showing the expected pattern of post-bond selective aggression23. The sham + C21 males also tended to decrease attack latency, but this decrease was not significant . In contrast, LS activation inhibited post-bond selective aggression; the latency to attack the intruder remained long in hM3 + C21 males when they were sex naïve and pair bonded . Moreover, LS-activated males (hM3 + C21) were slower to attack the intruder than hM3 + saline control males once they were pair bonded , and tended (non-significantly) to be slower to attack than pair bonded sham + C21 males .The latency to attack the intruder was affected by LS activation  = 9.41, p = 0.004; Fig. t35.3 = 2.71, p = 0.01; sham + C21: t35.3 = 2.05, p = 0.05). However, LS activation significantly inhibited pair bond-induced intruder chasing behaviors. That is, hM3 + C21 males did not differ in time spent chasing intruders when they were pair bonded compared to when they were sex naïve . Duration of pouncing did not yield any significant main effects  = 1.07, p = 0.35; bond status, F = 0.40, p = 0.53) or a significant interaction of treatment × bond status  = 0.69, p = 0.50).Subject males increased the instances of aggressive chases overall as they transitioned from sex naïve to pair bonded  = 10.63, p = 0.003; Fig. t35.3 = 2.75, p = 0.009; sham + C21: t35.3 = 2.78, p = 0.009). Unlike the two control groups, LS activation inhibited the normal expression of pair bond-induced aggressive behaviors in hM3 + C21 males . Taken together, these results indicate that LS activation inhibited selective aggression in pair bonded males.Finally, we examined how LS activation impacted the proportion of time subjects spent engaging in total aggressive behaviors . Linear mixed-model (LMM) analysis revealed a significant main effect of bond status  = 4.46, p = 0.02; Fig. t35.3 = 2.60, p = 0.01). In contrast, control males did not differ in their duration of huddling when they were sex naïve or pair bonded . Moreover, the duration of huddling in pair bonded LS-activated (hM3 + C21) males was significantly longer than pair bonded hM3 + saline control males , and pair bonded sham + C21 control males .The reduction of aggressive behaviors in LS-activated males was accompanied by an increase in social behaviors. Specifically, we found a significant interaction between treatment and bond status  = 21.81, p = 5.17 × 10−5; Fig. F = 4.19, p = 0.02). Post hoc analysis revealed that LS activation maintained the high instances of sex naïve anogenital sniffing after the bond was formed . Control males, however, demonstrated the anticipated reduction in prosocial anogenital sniffing with the intruder once they had established a bond .Anogenital sniffing, a common marker of prosocial investigation, during the RIT also showed a significant main effect of bond status  = 9.77, p = 0.004; Fig. t35.3 = −2.73, p = 0.0001; sham + C21: t35.3 = −2.51, p = 0.02). In contrast, LS activation maintained the expression of prosocial behaviors with a stranger after bonding . Taken together, results from comparisons of aggressive and prosocial behaviors in the resident-intruder paradigm indicate that LS activation interferes with the selective aggression typical of bonded male prairie voles, and actually promotes prosocial behaviors with intruders.Lastly, we examined how LS activation impacted the total time subjects spent engaging in social behaviors  when males were sex naïve and pair bonded. Our results demonstrated a significant main effect of bond status  are territorial, appear to mate guard, and become  selectively aggressive to conspecifics other than their partner and offspring. The reproductive success of wanderers is less than, or possibly equal to, that of their pair bonded resident counterparts34 suggesting that males benefit reproductively to a greater extent when bonded. Notably, the putative reproductive advantage for residents is directly associated with this behavioral shift, in which males go from being indiscriminately social and relatively non-aggressive, to being selectively social with partners and selectively aggressive to other adults. Our study provides insight into the mechanistic control of this specific behavioral shift, implicating the LS as a major node in the network that modulates these forms of social behavior. Specifically, we believe our data support the hypothesis that the LS functions to shift the balance between general affiliation and social selectivity in a context-dependent manner36. In this case, such a shift enables the behavioral transition associated with being single to being bonded.The transition from being a single (un-bonded) male to a pair bonded socially monogamous male is accompanied by a suite of characteristic changes in behavior. Single males (referred to as “wanderers” in nature) are non-territorial and typically inhabit expansive home ranges that intrude into the home ranges of many other conspecifics37, but mechanisms that control prairie vole bonding (including the actions of vasopressin and oxytocin within the LS15) seem to parallel the mechanisms that govern human social attachment38. The LS has long been appreciated as a central node within pair bonding neural circuitry13. Yet the role of the LS in this process has not been terribly clear.The neurobiology of pair bonding has benefited greatly from studies using prairie voles to uncover the specific mechanisms that modulate this important process. Pair bonding is rare among mammals39, where males exposed to females showed elevated Fos induction in the LS in comparison to males that never interacted with a female, irrespective of mating. Additionally, males that had prior experience with a female exhibited higher Fos staining in the LS in response to a male intruder relative to males that had no prior experience with a female39. At the time, it was difficult to determine the physiological significance of Fos induction in the LS . Evidence from our chemogenetic manipulations likely indicates that the LS was inhibited in the males that showed mating-induced aggression toward male intruders and Fos induction in the LS.One of the first pieces of evidence linking LS neural activation with reproductive history and mating-induced aggression in male prairie voles was demonstrated by Wang et al.40, which can be paired with highly valanced social reward during mating, thereby enabling animals to associate selective preferences for a particular partner over all others41. Indeed, the LS (and oxytocin and vasopressin acting therein) have been frequently associated with differences in social recognition and discrimination, supporting this interpretation. Furthermore, like the other behaviors just discussed, prairie vole social recognition is altered after a bond has formed. Zheng et al.42 showed that sex naïve adult male prairie voles fail to distinguish female conspecifics but effectively discriminate between males. This indiscriminate social recognition of females but clear ability to discriminate between males occurs when males are single. In nature, single males are described as wanderers—a pivotal period of life when it is necessary to be indiscriminate of mate choice  while taking note of and being equipped to avoid potentially aggressive resident males and wanderer competitors. Once males establish a bond and form territories, a shift in cognition occurs where they now discriminate between females43. The cognitive shift for social discrimination that is associated with pre- and post-bonding reproductive status presumably enables males to distinguish among conspecifics and act appropriately in prosocial (selective affiliation) or antisocial (selective aggression) interactions. Taken together, the LS is well positioned to enable numerous shifts in cognition and behavior that facilitate this key change in life-history stages.One hypothesis aimed at explaining the functional role of the LS in bonding is that it facilitates social recognition5 elegantly demonstrated that the medial bed nucleus of the stria terminalis (BSTm) and the LS modulate social grouping based on the impact of vasotocin . In fact, the neural expression and function of VT directly relates to sociality in solitary and gregarious species of Estrildid finches5. Manipulation of the BSTm-LS circuit in Estrildid finches alters social grouping behaviors, suggesting that VT impacts preferences for social grouping independently of mating system3. These results in birds strongly parallel our results in voles, in which DREADD-mediated LS activation eliminated selective aggression and promoted prosocial behaviors in both the social approach and resident-intruder paradigms, while leaving the pair bond intact. This important caveat highlights the notions that (1) the LS impacts some general aspects of prosocial and antisocial behavior independently of other specific forms of social affiliation , (2) that it has functions that are generally similar across species but also specific to the life-history of a particular species , or (3) both.The shift between prosocial and antisocial behaviors within prairie voles that we have reported is consistent with previous work that has focused on Estrildid finches, an all-monogamous genus of birds. Kelly et al.14, social attachment to mates44, social memory46, social fear during lactation47, juvenile play48, social approach17, and aggression50. These studies have beautifully demonstrated the modulatory role the LS plays in many discrete aspects of social behavior. For example, lesioning the LS and LS-GABAA receptor activation cause irritability and aggression in mice52 and hamsters54. Further, Clemmens et al.14 showed that lesioning the LS disrupts age-dependent kin preferences in young rat pups. Instead of lesioning or inactivating the LS, we took a complimentary approach to chemogenetically activate the LS and examine its effect on prosocial behaviors and aggression in sex naïve male prairie voles, and later after they have become pair bonded. Notably, suppression of pair bond-induced aggression and promotion of social behaviors via LS activation in male prairie voles is congruous with previous work using optogenetics to activate LS neurons in sexually experienced male mice during resident intruder testing6. Wong et al.6 demonstrated that optogenetic stimulation of the LS in male mice significantly decreases the latency to attack, decreases the duration of attack behaviors, and increases the amount of time spent investigating a male intruder. Moreover, when the intruder is a female, LS optogenetic activation suppressed aggression, decreased the latency to stop mounting, and reduced the duration of time spent mounting6.Yet, to assume that the LS is so specialized to control only social grouping behaviors or social recognition, for example, ignores the evidence that has implicated it in numerous forms of social behaviors and closely associated cognitive behaviors, such as kin preference36. Indeed, the LS might function to modulate behavioral responses appropriate to particular environmental stimuli or internal life-history states through its connections within a larger network of brain areas, due to its cellular heterogeneity, or both55. Several excellent candidate circuits have been characterized, most of which are directly or indirectly associated with the general control of social behavior59. We believe that selective DREADD-mediated activation of the LS in our study tapped into and disrupted the dynamics within such circuitry and cellular dynamics that normally function to enable prairie voles to adjust behavior appropriately to account for important life-history shifts necessary to maximize their reproductive success. In this way, we have potentially provided evidence supporting the notion that the LS modulates context-specific motivational states in prosocial and antisocial behavior that are dictated by bonding status.A broader view of the function of the LS is that it plays a key role in modulating context-specific motivational states2. Notably, the LS is critical for the expression of both monogamous pair bonding and sociability/aggression, suggesting that it could serve as a point of communication between the sub-networks that govern different forms of social behavior57 to facilitate the greater function of the social brain9. In prairie voles, the shift from general sociability to social selectivity is a natural consequence of mating, which serves to guard mates and is an essential element of pair bonding. To our knowledge this is the first study to examine how chemogenetic control of the LS can shift the balance between general affiliation and social selectivity in male prairie voles as they transition between being sexually inexperienced and generally affiliative, to forming a pair bond by displaying social selectivity and territorial aggression. Our study advances understanding of the potential role that the LS takes on altering both prosocial and antisocial behaviors during a life history transition in reproductive state within the same individuals.In sum, we leveraged the social bonding nature of prairie voles to examine the impact that the LS has on altering both prosocial and antisocial behaviors during a life-history transition from being single to being paired within the same individuals. Our study suggests that DREADD-mediated activation of the LS promotes prosocial behaviors and inhibits pair bonding-induced selective aggression, but does not affect the critical ability to form selective bonds and partner preferences. The LS may act as a nexus of prosocial and antisocial behaviors in male prairie voles, permitting physiological states and environmental demands to influence social phenotypes and reproductive decision-making. We previously argued that mating systems could be viewed as an independent “behavioral axis” that is orthogonal to sociabilityMale and female prairie voles used in this study were produced in our breeding colony at Cornell University, from breeding pairs that were offspring of wild caught animals captured in Champagne County, Illinois, USA. All subjects were unrelated, and sexually mature virgin males between 90 and 120 days old. Animals were weaned and housed with littermates on postnatal day (PND) 21, and then housed with same-sex littermates after PND 42-45. All animals received rodent chow  and water ad libitum, and were maintained under standard laboratory conditions  in transparent polycarbonate cages (46.5 × 25 × 15.5 cm) lined with Sani-chip bedding and provided nesting material. All experimental procedures were conducted and approved by the Institutional Animal Care and Use Committee of Cornell University (2013-0102) and were in accordance with the guidelines set forth by the National Institutes of Health.12 vg/ml in sterile 0.1 M PBS and stored in 5 µl aliquots at −80 °C until the day of use. Before surgery, male subjects were anesthetized with 1.5–2% isoflurane mixed with pure oxygen (1 l//min) and fixed in a stereotaxic apparatus (Kopf Instruments). The scalp area was scrubbed with povidone-iodine (Purdue Products), and ophthalmic ointment (Henry Schein) was applied to the eyes. Subjects received either bilateral injections of the excitatory DREADD or sterile 0.1 M PBS into the LS . The virus suspended in PBS or PBS alone was delivered at a volume of 300 nl/side using a 1.0 µl syringe  at a rate of 75 nl/min -mCherry excitatory DREADD (hM3 for short), a gift from Bryan Roth (Addgene plasmid # 50474), was diluted to 1 × 10min Fig. . Followi61. Thirty minutes before each behavioral test began , subjects were treated with an acute injection of either 3 mg/kg C21 (hM3 + C21) or saline . A third group of subjects received 0.1 M PBS injections into the LS to control for the injection of the hM3 DREADD viral vector and were acutely injected with 3 mg/kg C21 (sham + C21) 30 min before each behavioral test. This control group enabled assessment of potential C21-induced off-target effects in the absence of the hM3 receptor30. C21 was dissolved in sterile saline and injected intraperitoneally (i.p.) through a 26-gauge needle and 1 ml syringe. Sample sizes for each behavioral test are defined below.Sixteen days after administering DREADDs to the LS (day 0), all subjects underwent a SAT to measure sex naïve same-sex affiliation. Four days later (20 days post-surgery), animals were subjected to a partner preference test (PPT) to measure their ability to form pair bonds with opposite-sex conspecifics. Two RITs were administered to subjects to measure sex naïve aggression  and pair bond-induced aggression . Figure N = 32) were introduced and allowed to acclimate to the SAT apparatus (20 × 40 × 28 cm) for 30 min. After acclimation, a doorway separating the testing chamber from a stimulus presentation box (10.06 cm3) containing an unfamiliar, age-matched, same-sex conspecific was unblocked, exposing the subject to the stimulus male conspecific. The stimulus chamber was separated from the testing chamber with a wall containing 13 holes (1.27 cm diameter) allowing for visual, auditory, and olfactory contact between the two animals. Tests were video recorded for a 10 min trial and the duration in the social zone and non-social zone, latency to approach the stimulus chamber, zone transitions, distance moved, and velocity were quantified. We quantified the latency to social approach as the difference in time from the start of the test until the nose of the subject was within 3 cm of the stimulus chamber.Animals (N = 29) were pair-housed with sexually primed females for 48 h before the PPT was performed , comprised of a central “neutral chamber” (45.7 × 50.8 × 30.5 cm) and two “side chambers” (27.9 × 50.8 × 30.5 cm). Off-set doorways allow the stimulus animal unrestricted access to the entire apparatus. The female used for cohabitation  is secured to one side chamber and a novel female  is secured to the other side chamber using lightweight chains attached to zip-tie neck collars that are attached to the wall of each side chamber. Stimulus animals adapt quickly to the tethers and collars, and can engage in the full range of natural behaviors. Male subjects are allowed to freely explore the apparatus and interact with either of the two females for 3 h while being video recorded. A trained experimenter blind to the treatment groups quantified the contact duration between the subjects and both their partners and strangers. Contact duration was calculated as time spent huddling, sniffing, and grooming the partner or stranger. This measure was used to calculate the total contact duration and a partner preference index . A partner preference is defined as when the subject spends more time more time in side-by-side contact with the partner compared to the stranger13. Due to technical difficulties with recording equipment, two hM3 + saline subjects and one sham + C21 subject were excluded from analysis from the partner preference data analysis  were examined using the RIT on testing days 17 and 21, respectively  in PBS under deep anesthesia. Brains were extracted and post-fixed for 24 h in 4% PFA, cryoprotected in 30% sucrose for 48 h, then frozen and stored in cryoprotectant at −80 °C. Coronal sections (40 μm) containing the LS were cryosectioned (Leica Cryostat CM 1950) and collected for confirmation of DREADD expression via visualization of the mCherry fluorescent tag under a 10x objective (Leica DM550 B).64, emmeans65, and lmerTest66. Significant interactions or significant main effects (α ≤ 0.05) were followed by two-tailed Tukey’s post hoc test. For the PPT, a one-tailed (right sided) Tukey’s post hoc test was used because of our a priori assumption that subjects would exhibit a partner preference. For comparisons between treatment, data were tested for normality (Shapiro–Wilk) and equal variance. When data were found to be normally distributed, an unpaired t-test (two-tailed) was performed. If data were not normally distributed, a Wilcoxon rank sum test was performed. Figures were created using Prism version 9.0.1.151  and all data are presented as the means ± standard error of mean.All behavioral procedures were manually scored by an observer blind to treatment using Noldus Ethovision XT 13 , Noldus Observer XT 11, or BORIS 7.9.7. Data were analyzed with RStudio (version 1.2.1335) using a LMM framework with the packages lme4Behaviors from the SAT were analyzed to compare the effects of treatment between subjects  on zone duration , latency to social approach, zone transitions, distance moved, and velocity. Behaviors from the PPT were analyzed to compare the effects of treatment between subjects  on contact duration (partner vs. stranger), partner preference index, total contact duration, and cage transitions. Behaviors from the resident-intruder tests were analyzed to compare the effects of treatment between subjects  and bonding status within subjects (sex naïve vs. pair bonded) on social and aggressive behaviors.Further information on research design is available in the Supplemental MaterialDescription of Additional Supplementary FilesSupplementary Dataset 1Supplementary Dataset 2Supplementary Dataset 3Supplementary Dataset 4Supplementary Dataset 5Supplementary Dataset 6Supplementary Dataset 7Supplementary Dataset 8Supplementary Dataset 9Supplementary Dataset 10Supplementary Dataset 11Supplementary Dataset 12Supplementary Dataset 13Supplementary Dataset 14Supplementary Dataset 15Reporting Summary


N = 60) was randomly assigned to one of three conditions representing MIs across the warmth‐competence stereotype space. Patterns of self‐completed measures for stereotypes, emotions and behaviors differed significantly between conditions. The association between stereotypes and behaviors were largely mediated by emotions. Systematic patterns of stereotypic perceptions, emotional prejudices and behavioral discrimination are present for individuals with different types of MIs. Hence, generic measures of discrimination, such as social distance scales, may be misleading. Intervention strategies should consider the systematic variation of the factors involved in stigma, differentiating by type of MI and discriminatory behaviors.Mental illness (MI) stigma has been characterized as multi‐dimensional including problems of knowledge (stereotypes), attitudes (prejudice) and behavior (discrimination); however, most research practice is predominantly applying unidimensional behavioral measures such as social distance scales. Moreover, specific types of MIs and different forms of discriminatory behaviors are not being differentiated. The Stereotype Content Model predicts that group stereotypes  are linked with different forms of discriminatory behaviors (harm or facilitation) via emotional prejudices . The present study sought to establish how differential stereotypic perceptions of MI subgroups elicit distinct forms of behavioral discrimination via emotional prejudices. A community sample ( Furthering this understanding may open targeted pathways to reducing prejudice and discrimination towards individuals with MI. The Stereotype Content Model  and whether they are competent or able of enacting them (competence). As a result, two fundamental dimensions have been proposed to systematically explain the stereotypic perceptions of different social groups – warmth  and competence . Consequently, the content of the stereotypes about all social groups are represented as a combination of these two dimensions of warmth and competence. Four different quadrants of the warmth‐competence space have been identified: a positive quadrant of high warmth and high competence (HW/HC), a negative quadrant of low warmth and low competence (LW/LC) and two quadrants of mixed or ambivalent stereotypes , one low in warmth and high in competence (LW/HC) and one high in competence but low in warmth (HC/LW). Social structural variables are thought to causally predict the warmth and competence ratings of groups. People infer (low) warmth from competition (vs. cooperation) and competence from status. Numerous studies have confirmed the warmth‐competence space as well as their socio‐structural antecedents, for example, in representative samples  and elicits emotions of pride . The LW/LC quadrant contains societal outcasts  and is associated with emotions such as contempt and disgust. The two other quadrants, referred to as ambivalent, contain the LW/HC stereotype, which contains successful outsiders, such as entrepreneurs  and linked with emotions of envy and jealousy, as well as the HW/LC quadrant, which contains groups, such as the elderly or disabled, and elicits emotions of pity and sympathy Fiske, , 2018.et al., The BIAS map has been designed  was employed to probe transferability of previous findings across samples (Cuddy Hypothesis 1(a) social structures of high competition and low status;(b) stereotypic ratings of low warmth and low competence;(c) emotional prejudices of contempt; and(d) behavioral discrimination of active and passive harm.The schizophrenia subgroup (LW/LC), will be primarily associated with:Hypothesis 2(a) social structures of low competition (cooperation) and low status;(b) stereotypic ratings of high warmth and low competence;(c) emotional prejudice of pity; and(d) behavioral discrimination of active facilitation and passive harm.The Alzheimer's subgroup (HW/LC)will be primarily associated with:Hypothesis 3(a) social structures of low competition (cooperation) and high status;(b) stereotypic ratings of high warmth and high competence;(c) emotional prejudice of admiration; and(d) behavioral tendencies of active and passive facilitation.The control group (HW/HC)will be primarily associated with:Firstly, the current research sought to examine whether social structure, stereotypes, emotional prejudice and behavioral discrimination for each of the proposed subgroups would confirm the predictions of the SCM in line with their proposed relative location in the warmth‐competence space. That is, it was hypothesized that compared to the other subgroups:et al., Hypothesis 4Warmth (high) is hypothesized to predict active facilitation mediated via pity and admiration.Hypothesis 5Warmth (low) is hypothesized to predict active harm mediated via contempt and envy.Hypothesis 6Competence (high) is hypothesized to predict passive facilitation mediated via envy and admiration.Hypothesis 7Competence (low) is hypothesized to predict passive harm mediated via contempt and pity.Second, it was tested whether the proposed associations of stereotypes with behavioral discrimination via emotional prejudice would hold in line with the BIAS map  were recruited via systematic sampling from a retail center in a metropolitan area in the United Kingdom, that is, every 6th person passing by was approached for participation. The majority, 56.7% indicated to be White British, 16.7% Black/Black British, 16.6% Asian/Asian British, 6.7% White Other, and 3.3% mixed heritage. Participants were screened prior to being randomly allocated to one of the three conditions . Participants were taken into a quieter corner within the retail center and out of earshot from the regular footfall. This was facilitated by the questionnaire itself only taking a few minutes per participant to complete, ensuring that participant confidentiality was recognized. Upon completion of the questionnaire, participants were debriefed and handed information flyers on schizophrenia and Alzheimer's disease which included useful support contacts if needed. After completion, a chocolate bar was given to participants as a reward. Those reporting a personal or family history of mental health illness were excluded from the remainder of the study (n < 10). That is, a screening sheet was given, that was presented as a short demographic survey to the willing participants. Those affirming as having a historical background, or any personal experience of MI(es) were thanked for their participation, and given their chocolate bar as a reward upon the screening sheets completion. Ethical approval was obtained by ethics committee of the University of West London and participants provided informed consent prior to participation.Sixty individuals , passive facilitation , passive harm . Scales were tested for skewness and kurtosis ; hence, the inverse of the scale, multiplied by ‘−1’ to maintain the direction of effects, was used for the remainder of the analyses. A small number of participants (max. n = 3) did not complete all items of each scale. That is, one participant did not complete the status scale, two participants did not complete the competition scale and one participant did not complete the active harm scale. All analyses were conducted on complete data only. Given the small number of omissions, procedures for dealing with missing data were not considered.Questionnaires were similar across conditions except for the group that scale items referred to, that is, either the two MI subgroups  or the control group . Questionnaire items from research on SCM and the BIAS Map were used  = 8.57, p = 0.001, η2p = 0.24, and structure, F = 8.43, p = 0.005, η2p = 0.13, as well as a significant interaction effect, F = 5.27, p = 0.008, η2p = 0.16. Between‐group contrasts showed no significant differences for competition, all t(55)s < 1.3, however, the control group was perceived as significantly higher in status compared to both, the Alzheimer's and the schizophrenia subgroups, t(56)s = 4 and 4.69, ps < 0.001).A 3 (condition) by 2  analysis of variance (ANOVA) revealed a significant main effects for condition, F = 29.05, p < 0.001, η2p = 0.51, but not for stereotype, F = 1.75, p = 0.19, η2p = 0.03, however, there was a significant interaction effect, F = 22.07, p < 0.001, η2p = 0.44. Between‐group contrasts showed that all three groups differed significantly from one another in warmth, t(57)s = 2.92–6.46, all ps < 0.005) with Alzheimer showing the highest ratings, followed by the control and schizophrenia subgroups. Both, the Alzheimer's and the schizophrenia subgroups were also significantly lower from the control group in competence, t(57)s = 6.06 and 7.46, all ps < 0.001) whilst they did not differ from one another. Moreover, planned comparisons testing for ambivalent stereotypes (high on one dimension but low on the other) revealed that the Alzheimer's as well as the control subgroups showed a significant difference between warmth and competence, t(19)s = 3.50 and 4.74, ps < 0.001) whereby the Alzheimer's group was seen as higher in warmth and the control group as higher in competence.A 3 (condition) by 2 (stereotypes) ANOVA showed a significant main effect for condition, F = 6.01, p = 0.004, η2p = 0.18, and emotions, F = 40.85, p < 0.001, η2p = 0.42, as well as a significant interaction effect, F = 24.51, p < 0.001, η2p = 0.46. Between‐group contrasts showed that a higher amount of contempt was expressed towards the schizophrenia group when compared to the control as well as the Alzheimer groups, t(57)s = 4.97 and 6.14, ps < 0.001, more pity was expressed towards the Alzheimer group when compared to the control as well as the schizophrenia groups, t(57)s = 4.11 and 5.21 2.86–7.3, ps < 0.001, and lastly, the control group elicited both, more admiration as well as more envy, compared to the Alzheimer and the schizophrenia groups, t(57)s = 2.87–7.32, all ps < 0.005. Planned comparisons within subgroups revealed that pity was significantly higher for the Alzheimer's subgroup, t(19)s = 7.07–16.21, all ps < 0.001, and contempt was significantly higher for the schizophrenia subgroup when compared to the other three emotions, t(19)s = 4.26–13.03, all ps < 0.001. However, no significant differences between emotions were found for the control group, all t(19)s < 2.A 3 (condition) by 4 (emotions) ANOVA revealed significant main effects for condition, F = 1.58, p = 0.22, η2p = 0.05, however, significant effects were revealed for behaviors, F = 7.66, p < 0.001, η2p = 0.12, as well as the interaction effect, F = 20.41, p < 0.001, η2p = 0.42. Between‐group contrasts revealed that all groups differed significantly from one another on active harm with schizophrenia showing the highest ratings followed by the control and Alzheimer's groups, t(56)s = 2.32–5.28, all ps < 0.05. The schizophrenia group was also significantly higher in passive harm compared to the Alzheimer's as well as the control groups, t(56)s = 3.61 and 4.53, ps < 0.001. All groups differed significantly from one another on active facilitation with Alzheimer's showing the highest ratings followed by the control and schizophrenia groups, t(56)s = 2.21–6.00, all ps < 0.05. All groups also differed significantly from one another on passive facilitation with the control group showing the highest ratings followed by the Alzheimer's and schizophrenia groups, t(56)s = 2.67–6.85, all ps < 0.01. Planned comparisons within subgroups revealed that for the Alzheimer's subgroup the two expected behavioral tendencies were significantly higher than others, insofar as active facilitation was significantly higher than active harm and passive facilitation, whilst passive harm was significantly higher than active harm whilst additionally active facilitation was significantly higher than passive harm, t(19)s = 2.21–6.42, all ps < 0.05. For the schizophrenia subgroup also the two expected behavioral tendencies, active harm and passive harm, were significantly higher than all other behavioral tendencies whilst passive harm was significantly higher than active harm in addition, t(19)s = 2.65–7.42, all ps < 0.01. For the control group, one of the expected behavioral tendency, active facilitation, was not significantly higher than any other whilst the other expected behavioral tendency, passive facilitation, showed to be significantly higher than active harm as well as the expected behavior of active facilitation, t(19)s = 3.09–4.28, all ps < 0.01.A 3 (condition) by 4 (behaviors) ANOVA showed no significant effect for condition, et al., R2, see below for each model). Sufficient power was provided for each of the analyses  which was mediated by pity, total indirect effect = 0.13, 95% CI = , R2 = 0.44  which was mediated by contempt, total indirect effect = −0.14,, 95% CI = , R2 = 0.33 , which was mediated by both envy and admiration, total indirect effect = −0.29, 95% CI = , R2 = 0.66  was found, this was neither mediated via contempt nor pity, total indirect effect = −0.05, 95% CI = −0.22, 0.06], R2 = 0.38 .Except for passive harm, one emotion or both significantly mediated the direct effect of the stereotype on the behavioral tendency see Fig. . Warmth .44 Fig. . Warmth .33 Fig. . Compete.66 Fig. . Finally.38 Fig. ; howeveret al., et al., The present research explored how differential stereotypic perceptions of MI subgroups elicit differential emotional prejudices and behavioral discrimination. The work built on previous studies applying the SCM  and schizophrenia (low) as well as for the Alzheimer's group to emerge as an ambivalent stereotype (HW/LC). Against predictions, no group differences in competition emerged. Mirroring the unexpected findings for competition (the antecedent of warmth), the control group was unexpectedly seen as lower on warmth than the Alzheimer's group and emerged as an ambivalent stereotype (LW/HC) due to lower than expected warmth ratings. Given the high prevalence of MI in the population of about 1 in 3  when comparing between groups as well as between emotions. When comparing between groups, the control group showed the expected emotion of admiration; however, unexpectedly envy also emerged. Envy is associated with ambivalent stereotypes in the LW/HC quadrant . This finding was substantiated by the control group receiving lower ratings for active facilitation and higher ratings in active harm compared to the Alzheimer's group. It appears that in terms of the warmth‐competence space the lower than expected warmth ratings for the control group may have moved it from the expected HW/HC quadrant towards the LW/HC quadrant, which subsequently enhanced its association with envy and decreased behavioral tendencies for active facilitation whilst increasing those of active harm.All behavioral tendencies were associated with the stereotypes as predicted and, with the exception of passive harm, were mediated via the respective emotions put forward by the SCM; confirming that emotions were shown to be a stronger predictor of behavioral tendencies than stereotypes. The missing mediation as well as lack of association of contempt and pity with passive harm is puzzling. In particular, as both MI subgroups did show the predicted dominance of the behavioral tendency passive harm, along with the emotional responses . Although, contempt was significantly correlated with passive harm, warmth (entered as a control in the mediation) was correlated with both, contempt and passive harm, and may have overridden the effect of contempt in the mediation. This may have been exacerbated by the fact that passive harm ratings for the Alzheimer's group were slightly lower than expected (see above), further explaining the lack of associations with pity, the dominant emotion for this group.One of the main limitations evident in the results of the current study is the portrayal of the control group. The control group was intended to serve as a prototypical ingroup but, probably due to its description in terms of lack of a MI instead of representing a societal ingroup in its own right, it was seen to show somewhat lowered ratings in warmth, triggering the emotion of envy and passive but not the anticipated active facilitation, along with general undifferentiated feelings towards this group. Future research in this area should ensure the use of a label which can serve as a true prototypical ingroup.et al., A further limitation is the small number of MI subgroups used in this study. The current study was a first step in exploring the systematic patterns put forward by the BIAS for MI subgroups. The current selection of MI subgroups was based on their stability across previous studies in terms of their location in the warmth‐competence stereotype space and therefore to represent other MIs in these two main clusters  thereby suggesting that those findings may be more generalizable.et al., et al., et al., et al., The study's findings suggest that discriminatory behavioral responses systematically differ by type of MI thereby implying that generic measures of discrimination, such as social distance scales, may be misleading as they may not be able to capture the full range of discriminatory behaviors (Corrigan, et al., et al., et al., et al., et al., Emotional prejudices can take on the forms of contempt, pity or envy and accordingly discriminatory behaviors can vary from overt attacks to social exclusion or paternalizing behaviors. All of which can negatively impact on the well‐being of individuals with MI as well as their economic and social burden on society as a whole (Corbière This article builds on the second author's Undergraduate dissertation at the University of West London Ryan, .Ethical approval was obtained by the Ethics Committee at the University of West London and participants provided informed consent prior to participation.


Glioma stem cells (GSCs) play an important role in glioma recurrence and chemo‐radiotherapy (CRT) resistance. Currently, there is a lack of efficient treatment approaches targeting GSCs. This study aimed to explore the potential personalized treatment of patients with GSC‐enriched gliomas.Single‐cell RNA sequencing (scRNA‐seq) was used to identify the GSC‐related genes. Then, machine learning methods were applied for clustering and validation. The least absolute shrinkage and selection operator (LASSO) and COX regression were used to construct the risk scores. Survival analysis was performed. Additionally, the incidence of chemo‐radiotherapy resistance, immunotherapy status, and tumor treating field (TTF) therapy response were evaluated in high‐ and low‐risk scores groups.Two GSC clusters exhibited significantly different stemness indices, immune microenvironments, and genomic alterations. Based on GSC clusters, 11‐gene GSC risk scores were constructed, which exhibited a high predictive value for prognosis. In terms of therapy, patients with high GSC risk scores had a higher risk of resistance to chemotherapy. TTF therapy can comprehensively inhibit the malignant biological characteristics of the high GSC‐risk‐score gliomas.Our study constructed a GSC signature consisting of 11 GSC‐specific genes and identified its prognostic value in gliomas. TTF is a promising therapeutic approach for patients with GSC‐enriched glioma. We divided glioma patients into the high‐ and low‐glioma stem cell (GSC) groups. GSC‐enriched patients had higher chemo‐radiotherapy resistance and worse prognosis than low‐GSC patients. Tumor treating field (TTF) therapy can effectively inhibit malignant biological characteristics of GSC‐enriched gliomas, including mitosis, angiogenesis, migration and invasion potential. TTF also could break the original DNA balance in GSC‐enriched gliomas. Taken together, TTF is a promising therapeutic approach for GSC‐enriched glioma patients. Self‐renewal endows GSCs with differentiated progenies that constitute the bulk of glioma mass.2+ influx; and transforming membrane permeability.Tumor treating fields (TTFs) is a novel noninvasive antitumor treatment that uses low‐intensity, intermediate frequency, and alternating electric fields.Single‐cell RNA sequencing (scRNA‐seq) provides an encouraging method to identify marker genes of specific cells. In this study, scRNA‐seq was used to identify GSC‐related marker genes. Accordingly, machine learning methods, including non‐negative matrix factorization (NMF) and random forest (RF), were used to divide and validate patient clustering by referring these genes. Subsequently, least absolute shrinkage and selection operator (LASSO) regression and Cox regression were performed to construct risk scores, using the differently expressed GSC genes between the stratified groups. Finally, we applied GSCs risk scores to predict TTFs therapeutic sensitivity comparing to the chemo‐radiotherapy resistance and immunotherapy. This study expectantly developed personalized therapeutic approaches of TTFs targeting GSCs and served as a basis for future research on GSCs in gliomas.22.1https://www.ncbi.nlm.nih.gov/geo/) and CGGA (http://www.cgga.org.cn/index.jsp) databases, separately. The single‐cell transcriptome was used to screen for GSCs marker genes. The bulk tumor transcriptome data and clinical information of 698 and 1018 glioma samples were downloaded from The Cancer Genome Atlas  and CGGA databases, separately. Bulk tumor transcriptome was performed to divide subgroups based on marker genes. Of the 1728 patients enrolled in our study, GEO and TCGA data were used as the training set and CGGA data were used as the validation sets. To further investigate the genetic features of the subgroups, somatic mutations and somatic copy number alternations (CNAs), which corresponded to the patients with bulk tumor transcriptome data, were downloaded from the TCGA database. Bioinformatics analysis in our study utilized public databases, where the original studies obtained ethical approval. Informed consent was obtained from all the patients.Single‐cell transcriptome files of four glioma samples from GSE84465 and eight glioma samples from Chinese Glioma Genome Atlas (CGGA) scRNA‐seq were downloaded from the Gene Expression Omnibus  and “harmony” (version 0.1.0).2.3p‐value, precision, and F‐measure.Based on 146 intersected maker genes extracted from scRNA‐seq, the NMF algorithm was applied to identify robust clusters of TCGA patients.2.4The stemness indices of TCGA glioma samples were obtained from Malta's research.2.5The Xcell algorithm was performed to evaluate the enrichment level of 64 types of immune and stromal cells.2.6https://cloud.genepattern.org/). The variant types, variant classifications, and single‐nucleotide polymorphism were compared. Significant tumor‐mutated genes  across the two clusters were identified using the MutSigCV algorithm.To investigate the genetic features of the two clusters, arm‐ and focal‐level somatic CNAs were calculated using the GISTIC 2.0 analysis (2.7p < 0.001) between the two clusters. The GSC risk score of each patient was constructed by weighting the LASSO penalty coefficient. The median value of risk scores was set as the cutoff to divide glioma patients into low and high risk.LASSO and univariate COX regressions were applied to identify the differentially expressed GSC genes with non‐zero penalty coefficient and prognostic significance  curves were used to evaluate the predictive accuracy and performance of the nomogram model.2.950 value represented higher sensitivity.The R package “oncoPredict” was conducted to predict patient sensitivity to temozolomide (TMZ), and a lower ICIn addition, we also correlated the GSC risk score and TMZ sensitivity in our previously proposed glioma model integration system (patient‐derived glioma organoids and xenografts).2.10Neoantigens and six types of immune subtypes, obtained from Thorsson's research,2.11TTF therapy sensitivity was evaluated by five malignant biological features of gliomas: mitosis, angiogenesis, DNA repair, DNA damage, migration, and invasion.2.12Written informed consent was obtained from all patients. The study methodologies were approved by the Ethics Committee of Xiangya Hospital, Central South University . Patients enrolled in the trials received TTFs (150 kHz for >18 hours/day) by ASCLU‐300 TTF, which was designed by Hao Wu et al. and has been approved by the China Institute of Food and Drug verification .2.132 test and Fisher's exact test were used to compare the two groups. Spearman's correlation was performed to calculate correlation coefficients between continuous variables. Data were visualized using the R package “ggplot2”. Heatmap was generated based on the R package “pheatmap”. Survivorship curves were drawn with the R package “survminer”. NS indicated not statistically significant; *p < 0.05; **p < 0.01; ***p < 0.001. p < 0.05 was considered statistically significant.Statistical analysis was carried out using R software (version 4.1.2). Machine learning was performed by Weka software (version 3.8.4). Kaplan–Meier curves with the log‐rank test were used to compare the survival rates between the two groups. Univariate and multivariate Cox regression was applied to explore prognostic factors. Normal distribution was examined by Shapiro–Wilk normality tests. For continuous variables, the two‐tailed t‐test  and Mann–Whitney test  were used to compare the two groups. For categorical variables, χ33.1The design flow chart of this study was presented in Figure 3.2F‐measures of the grouping were 0.935, 0.734, and 0.725 in TCGA, CGGA325, and CGGA693, separately  with wild‐type IDH and a methylated MGMT promoter. The patient received standard radiotherapy and concomitant chemotherapy. TTF therapy was administered 4 months after surgery. No recurrence was detected on routine surveillance scans until 12 months after surgery Figure . Patient3.7In addition, we investigated the tumor microenvironment of the two clusters using Xcell, CIBERSORT, and estimate algorithms. Xcell algorithms showed the disparity of 64 immune and stromal cell types between the two clusters Figures  and S8. The clinical traits of the two clusters were also different. The cluter1 had significantly higher pathological grades than the cluster 2 Figure . Samples3.8Somatic mutations and CNAs were investigated between the two clusters, based on the TCGA dataset. Cluster1 had significantly higher arm‐and focal‐level amplification frequencies in chromosomes 7p and 7q and higher deletion frequencies in chromosomes 10p and 10q Figures . MutatioThe Waterfall plot showed mutation landscapes with significantly different frequencies in the two clusters Figure . Cluster3.9Considering the apparent disparities in the immune microenvironment between the two clusters, immunotherapy that targets GSCs was investigated. The high‐risk group exhibited higher expression levels of immune checkpoints, including antigen presence, cell adhesion, co‐stimulator, ligand, receptor, co‐inhibitor, and others Figures . Moreove4As the “ethnic minority” of glioma‐initiating cells, GSCs exerts its crucial role in chemo‐radiotherapy resistance. During chemo‐radiotherapy procedure, pGSCs can be transformed into mGSCs that demonstrates stronger resistance.Currently, in an in silico study, we novelty initiated and extracted the GSC‐related marker genes in gliomas using scRNA‐seq. Based on these marker genes, machine learnings were adopted for patient grouping and validation. An extended annotation of the stemness index, function enrichment, immune microenvironment, and genomic alterations was carried out for GSC‐related patient groups. Risk scores, based on the differentially expressed GSC genes between GSC‐stratified groups, were generated by Lasso and Cox regressions. Then, the risk scores were used to construct a nomogram prognostic model. Furthermore, chemo‐radiotherapy resistance, immunotherapeutic response, and TTF sensitivity were predicted based on the GSC risk scores.The two GSCs clusters had distinct biological features. Patients in cluster 1 with worse prognosis had higher proportions of MGMT promoter unmethylation, 1p19q noncodeletion, IDH wildtype, CL and ME subtypes, all of which correlated with the malignant phenotype. Besides, cluster1 had higher stemness indices, which indicated increased GSCs enrichment. Notably, despite the relatively high immune scores, patients in cluster 1 had higher expression of the immune checkpoint molecule programmed death‐ligand 1 (PD‐L1) and immunosuppressive cells, such as regulatory T cells (Tregs) and M2 macrophages. Previous studies have found that GSCs could recruit anti‐inflammatory M2 macrophages and induce Treg expansion to suppress both innate and adaptive immune responses.Based on the profiles of differentially expressed GSCs genes between the two clusters, we calculated GSCs‐related risk scores. The risk scores exhibited high efficiency to predict glioma patients' survival outcomes. Among the 11 genes with risk scores, ANXA1, encoding a membrane‐localized protein, plays an important role in stem cell maintenance and growth.GSCs’ resistance to chemoradiotherapy was increasingly recognized. GSCs overexpressed ATP‐binding cassette transporters (ABC) and MGMT to achieve multidrug resistance.TTFs is an emerging treatment in the field of tumor treatment, with a low side‐effect and resistance. Different from the systematic toxicity of chemotherapeutics, the side‐effect of TTFs mainly focused on local adverse skin effects, like dermatitis, erosin, and others.TTFs can effectively inhibit glioma growth and improve the survival outcomes, especially for the GBM.Therefore, we boldly speculated that TTF is a promising therapy targeting GSC‐enriched gliomas. TTFs can prevent the occurrence and progression of GSCs‐enriched gliomas by inhibiting mitosis, angiogenesis, migration, and invasion. Besides, TTFs disrupted the original DNA balance in GSC‐enriched gliomas by decreasing DNA repair and increasing DNA damage Figure .Considering the apparent disparities of the immune microenvironment between the two GSCs clusters, we also explore the immunotherapy response. Although, some immunotherapies have been identified to target GSCs,Although our study initiated some innovative perspectives, its limitations urgently require further evaluations. One major limitation was the lack of external real‐world data to confirm and support our findings. Another limitation was the lack of large glioma cohorts to predict the TTFs therapy response. In addition, the detailed mechanism of GSCs in TTF responses remained to be elucidated.5Our study constructed a GSCs signature consisting of 11 GSCs‐specific genes and identified its prognostic value in gliomas. Our results proved that TTFs was a promising therapeutic approach for GSCs‐enriched glioma patients.B.Chen, LY Zhang and CT Li designed and drafted the manuscript, TTF clinical organization; B. Chen, LY. Zhang and CT Li organized figures and edited legends. B Chen, XX Zhou, LT Yang, HS Zhou, M Meng and CT Li revised the article; B. Chen, LY. Zhang and CT Li conducted data analysis; XX Zhou, CT Li, H Wu, and ZX Liu organized patients with the treatment of TTF.This work was supported by the Nature Science Foundation of China (no. 81402249 to L.Y. Zhang), the Natural Science Foundation of Hunan Province (no. 2019JJ50963 to L.Y. Zhang), and Fundamental Research Funds for the Central Universities of Central South University (no. 160171016 to B. Chen).All authors declare no conflict of interest.Figure S1Click here for additional data file.Figure S2Click here for additional data file.Figure S3Click here for additional data file.Figure S4Click here for additional data file.Figure S5Click here for additional data file.Figure S6Click here for additional data file.Figure S7Click here for additional data file.Figure S8Click here for additional data file.Figure S9Click here for additional data file.Figure S10Click here for additional data file.Figure S11Click here for additional data file.Figure S12Click here for additional data file.Figure S13Click here for additional data file.Figure S14Click here for additional data file.