N = 2,921), we found the German FFNI to align with both, two- and three-factor models. The factors display good criterion validity with other narcissism measures, (non-)clinical personality dimensions, interpersonal styles, and adaptive adjustment. Neurotic and antagonistic narcissism discriminated between individuals with/without mental disorder diagnoses, and displayed a characteristic profile in incarcerated offenders. Since the FFNI is comprehensive but long, we constructed a 30-item brief form (FFNI-BF) optimizing the internal structure and external validity using ant colony optimization. The FFNI-BF displayed good psychometric characteristics and similar, in certain aspects even advantageous criterion validity. We conclude that the German FFNI validly measures key aspects of narcissism, and the FFNI-BF captures these in a concise manner.Narcissism is a multifaceted construct commonly conceptualized as comprising grandiose and vulnerable aspects in a two-factor model. While the manifold correlates of these aspects imposed a challenge for research on the structure of narcissism, recent models converge in a three-factor structure of agentic-extraverted, antagonistic, and neurotic aspects, capturing variance in different conceptualizations and correlates of narcissism. We construct and validate a German adaptation of the Five-Factor Narcissism Inventory (FFNI), a measure assessing these aspects based on the Five-Factor Model. In eight samples ( Narcissism has fascinated humankind for a long time, putatively because of its manifold consequences for our individual and social lives. In the ancient myth, Narcissus rejects the love of Echo, and is therefore condemned to fall in love with his mirror image, which ultimately leads him to a tragic death. This myth can be seen to reflect two main themes which are evident across a wide range of psychological literature on narcissism, namely grandiosity and vulnerability . The graDiagnostic and Statistical Manual of Mental Disorders . Despite their opposing experiential and behavioral tendencies, Wink noted both share “common narcissistic characteristics of conceit, self-indulgence, and disregard for the needs of others” (p. 596), thereby already anticipating contemporary personality models (see below). Building on this distinction, Complemental to the study of grandiose narcissism, vulnerable narcissism was studied in terms of a largely independent trait in the past decades . While tAs outlined above, both grandiose and vulnerable narcissism draw upon core narcissistic features such as self-importance, but, beyond that can be characterized by other trait constellations. In their Narcissism Spectrum Model,  The model is consistent with the process-oriented NARC model assuming self-enhancing self-regulatory strategies (agentic narcissism) and self-protecting self-regulatory strategies  was constructed as a narcissistic variant of the FFM altruism facet (agreeableness); a corresponding item is “it may seem unfair, but I deserve extra ” (authoritativeness facet (agentic narcissism) was constructed as a narcissistic variant of the FFM assertiveness facet (extraversion); a sample item is “I am comfortable taking on positions of authority.” Altogether, the FFNI comprises 15 facets which were selected based on prior expert ratings and meta-analyses (see The FFNI  is the tewards)” . Similaryses see . In addiyses see . Most reyses see .r = .74) with other trait measures of grandiose narcissism such as the NPI (r ≤ .60) such as the Minnesota Multiphasic Personality Inventory  and the vulnerable subscale of the Pathological Narcissism Inventory  and the HSNS  and vulnerable (.70 ≤ r ≤ .82) subscales of the PNI  construct a German version of the FFNI, (b) test its factor structure at different hierarchical levels in a confirmatory framework, and (c) corroborate and extend its nomological network regarding different aspects of personality and psychological functioning, (d) complemented by analyses of clinically relevant criteria and groups.The FFNI has been adapted for the use in Italian , NorwegiAs our third aim, we replicate and extend findings on the nomological network of the FFNI using other narcissism scales, measures of neighboring socially aversive personality traits, general FFM measures, as well as measures of interpersonal experience and behavior and adaptive and maladaptive psychological functioning. The latter are complemented by analyses of FFNI profiles related to clinically relevant characteristics (prior diagnoses/treatment) and groups (forensic sample).On the basis of the reviewed literature, we hypothesize that the FFNI two- and three-factor model scores will show substantial convergent validity with measures of grandiose and vulnerable narcissism  and agentic as well as antagonistic narcissism . We further hypothesize to find substantial convergence with the PNI, though in this case, we expect stronger alignment of the vulnerable than the grandiose dimensions, as PNI grandiosity, on a conceptual basis, puts stronger weight on maladaptive aspects . Both, gThe FFNI three-factor model dimensions should show substantial associations with the respective dimensions from general FFM inventories. Based on the literature, higher correlations can be expected for clinical than nonclinical FFM inventories. However, this concerns only the PID-5 dimensions of antagonism and neuroticism, as the PID-5 intro-/extraversion dimension emphasizes maladaptive introversion , hubristic pride , a cold-domineering interpersonal style (particularly its antagonistic aspects), game-playing and obsessive love styles, and measures of anger and aggression (particularly its antagonistic aspects). Antagonistic aspects should further relate to attachment anxiety and avoidance, while no particularly strong associations are expected for overall grandiose narcissism. Grandiose narcissism, particularly its agentic aspects, should relate positively to measures of psychological adjustment  and negatively to measures of maladjustment/psychopathology. Vulnerable narcissism should be associated with avoidance orientation (particularly neuroticism), hubristic pride, a wide array of interpersonal problems, game-playing and obsessive love styles, anger, and aggression (mainly antagonistic aspects), and attachment anxiety and avoidance (particularly neurotic aspects). It should correlate negatively with adaptive adjustment (self-esteem and life satisfaction) and positively with maladjustment . These correlations should be mostly due to neurotic, but also antagonistic aspects.We complement validity analyses of dimensional measures of maladaptive adjustment by more objective clinical and forensic criteria. For this, we assessed prior diagnoses in two of the included samples and administered the German FFNI in a sample of incarcerated young criminal offenders. Based on the literature, we expect that individuals with prior diagnoses will show elevated scores on vulnerable narcissism, particularly its neurotic aspects, and criminal offenders will show elevated scores on antagonistic narcissism. None of our hypotheses were preregistered. We report how we determined our sample size, all data exclusions, all manipulations, and all measures in the study.N = 2,921 participants  with a mean age of 25.59 years (SD = 7.67). Among those, N = 1,823 completed the long, and N = 1,098 the short FFNI. All samples were convenience samples assessed online, with the exception of sample 8, which includes young criminal offenders incarcerated in a correctional institution in the German state of Mecklenburg-Vorpommern. All data were collected in accordance with the relevant local guidelines and regulations and with the Declaration of Helsinki. There were no missing data (incomplete data sets were excluded a-priori). Beyond the scales listed in We gathered data from eight samples of German-speaking participants, four of which completed the long, and four of which the short version of the FFNI alongside different validity measures. The rationale was to pool several samples to obtain the largest possible sample size for analyses of the factorial structure and validity of the FFNI . Altogether, the samples comprised N = 1,745 (59.74%) of participants were students, 475 (16.26%) were employed, 50 (1.71%) were apprentices or secondary / high school pupils, 90 (3.08%) retired or unemployed, and the 68 (2.33%) participants of sample 8 were incarcerated . Among the pooled sample, indifference, exhibitionism, authoritativeness, grandiose fantasies, manipulativeness, exploitativeness, entitlement, lack of empathy, arrogance, acclaim seeking, thrill seeking) and vulnerable narcissism , antagonistic narcissism , and neurotic narcissism /60 items (FFNI-SF) belonging to 15 facets which were constructed as narcissistic variants of FFM facets : https://osf.io/ae7vr/The FFNI was initially translated into German independently by several members of the research groups from Bern and Münster. Next, a final consensus version was created by three psychologists all experts in narcissism and one of whom is a bi-lingual native speaker. This process involved an iterative approach in which each translated item was evaluated and in case of deviant translations the best wording was determined through discussion until consensus was reached. Following this, a back-translation was performed by a different bi-lingual native speaker, upon which any discrepancies again were resolved through discussion. Items of the German FFNI are available in This section provides information on the validity measures included in this study. leadership/authority and grandiose exhibitionism) and antagonistic narcissism  and antagonistic  aspects of grandiose narcissism . The admThe HSNS is a 10-item measure of vulnerable narcissism covering its antagonistic and neurotic aspects , primitive defense (reactions to self-threats such as projection or splitting), and impaired reality testing , ludus (game-playing love), storge (friendship love), pragma (pragmatic love), mania (obsessive love), and agape .The LAS  assessesWe used the 10-item RSES  for the We used the five-item SWLS  as an inWe used the 53-item BSI  to assesWe used the German version of the CES-D , the 20-N = 1,823 participants who filled in the FFNI-LF and N = 1,098 who filled in the FFNI-SF. Sample sizes for validity analyses differ depending on which inventories were included in the samples .Our rationale was to conduct analyses based on the largest set of available data across the samples. We pooled samples for analyses of reliability and factor structure for the different FFNI versions, leading to pooled data of ples see  and 4. F Here, the first-order factor loadings are first estimated in ESEM. These loadings (including cross-loadings) are then used as constraints for an estimation of the higher-order model in CFA. As not all item-level data were normally distributed, we attempted to estimate models using weighted least square mean and variance adjusted (WLSMV) estimators. However, many of the models could not be estimated using WLSMV (nonconvergence); therefore, we used maximum likelihood (ML) estimation for CFA and ESEM models instead.The analysis proceeded in three steps—descriptives and internal consistencies, confirmatory examination of higher- and lower-order factor-structure, followed by convergent and discriminant validity. We first analyzed internal consistencies and intercorrelations of the German FFNI in its different versions (including the brief form presented in Study 2) at the two-factor, three-factor, and facet level. Next, we set up confirmatory factor analysis (CFA) models to analyze the factor structure at the different levels . As the FFNI items display substantial cross-loadings, we complemented traditional CFA models with exploratory structural equation models ESEMs; . ESEMs cFinally, we analyzed correlations with validity measures assessing narcissism and related constructs, nonclinical and clinical personality constructs, interpersonal experience and behavior, as well as psychological adjustment and maladjustment.The intercorrelations among the scores according to the two- and three-factor models were generally as expected and similar for the different scale versions: grandiose and vulnerable narcissism displayed no or slight positive associations  displayed the best data fit. Here, we observed good fit for the FFNI-LF in both the two-factor  and three-factor solution . Also, the FFNI-SF displayed good fit in the two-factor  and three-factor solution . Regarding the FFNI-SSF, ESEM models with cross-loadings of items on factors for the FFNI-SSF displayed largely acceptable fit in the two-factor solution  and good fit in the three-factor solution .Taken together, though many of the models did not show acceptable fit by conventional standards, hierarchical CFA and ESEM models did so for different versions of the FFNI. Fit of ESEM-within-CFA models was generally superior to those of traditional higher-order CFA models, indicating that considering cross-loadings at item-level improves model fit. However, this also shows that the item sets of existing versions might not be optimally suited to capture the intended structure in an unidimensional manner. While ESEM models can effectively capture variance related to different aspects at item-level, we also note that they may run at the risk of over-fitting the data . In Study 2, we thus developed a novel short form by selecting a set of items which best represented the intended structure of the FFNI in terms of model fit and factor loadings.indifference and arrogance did not load significantly on the grandiose narcissism factor, and the ESEM-within-CFA model for the three-factor solution (Model 14) in the FFNI-LF, in which the facets arrogance, reactive anger, and distrust did not load significantly on antagonistic narcissism. This indicates that considering cross-loadings at item-level, though improving model fit, unveils problems in the factorial structure of the FFNI-LF, in the way that these facets do no longer represent the designated higher-level factors . Such problems were not evident in the corresponding models of the FFNI-SF, where all facet loadings were significant, showing that shortened versions of the FFNI are better suited to capture the intended factor structure.Turning to factor loadings , and—to a smaller extent—the PNI grandiose scales. Correlations with vulnerable narcissism measures  were low to moderate. FFNI vulnerable narcissism, on the contrary, was not generally associated with those narcissism scales assessing primarily agentic aspects of grandiose narcissism , but was associated with antagonistic scales . FFNI vulnerable narcissism was strongly and consistently associated with other measures of vulnerable narcissism. Both, FFNI grandiose and vulnerable narcissism were related to the DAPP as a clinical narcissism scale (though vulnerable narcissism somewhat stronger), and both were associated with psychological entitlement, as predicted by theory . Within the three-factor model, as expected, FFNI agentic narcissism was primarily and strongly associated with other agentic narcissism scales, but also antagonistic narcissism scales, and displayed low-to-moderate associations with measures of vulnerable narcissism. FFNI antagonistic narcissism was primarily associated with other antagonistic narcissism scales, though also with agentic narcissism scales, substantially associated with vulnerable narcissism scales, and psychological entitlement. FFNI neurotic narcissism was slightly negatively associated with agentic narcissism scales, partially associated with antagonistic scales , and strongly associated with vulnerable narcissism scales . Associations with psychological entitlement were low to negligible. Regarding the Dark Triad traits, as assessed with the SDT, FFNI grandiose narcissism, particularly its agentic aspects, displayed the strongest convergent associations with SDT narcissism , whereas antagonistic aspects were more strongly related to SDT Machiavellianism and psychopathy.In the NEO-PI-R as a general FFM personality measure, FFNI grandiose narcissism reflected—as expected—primarily in an extraverted and disagreeable personality profile. We also observed negative correlations with facets of neuroticism as well as sporadic associations with openness and conscientiousness (particularly achievement striving). Vulnerable narcissism—as expected—reflected primarily neuroticism and disagreeableness (though the latter was not as pronounced in the SSF). Vulnerable narcissism was further considerably associated with introversion and low conscientiousness. Within the three-factor model, it is interesting to note that though FFNI agentic narcissism did show the expected associations with NEO extraversion; relations with disagreeableness were also prominent. We further observed substantial positive associations with openness and conscientiousness. FFNI antagonistic narcissism was mainly and consistently associated with disagreeableness , and FFNI neurotic narcissism was strongly and consistently associated with NEO neuroticism, and also considerably associated with introversion. Interestingly, FFNI neurotic narcissism was slightly positively associated with some aspects of agreeableness.Consistent with predictions, grandiose narcissism—particularly its agentic aspects—was associated with behavioral activation in the BIS/BAS, whereas vulnerable narcissism—particularly its neurotic aspects—was associated with behavioral inhibition. Grandiose narcissism was related mainly to hubristic but also authentic pride. Closer differentiation in the three-factor model shows that associations with hubristic pride is mostly due to antagonistic narcissism. Vulnerable narcissism—particularly neurotic narcissism—was negatively associated with authentic pride.Within the clinical FFM dimensions as assessed by the PID-5, FFNI grandiose narcissism was mainly associated with antagonism. Interestingly, also psychoticism and disinhibition displayed positive associations. Vulnerable narcissism, as expected, displayed the strongest associations with negative affect, but correlated with all of the PID-5 dimensions . Inspection within the three-factor model shows that it is particularly FFNI antagonistic narcissism which shows associations to all PID-5.DSM-based measure of borderline personality disorder (MSI-BPD), which also reflects in substantial correlations of borderline personality disorder and vulnerable and, to some extent, grandiose narcissism. All FFNI narcissism dimensions displayed substantial associations with the IPO as a measure of general personality functioning. Antagonistic narcissism was more related to primitive defenses and impaired reality testing, whereas neurotic narcissism was more related to identity diffusion.FFNI antagonistic narcissism and vulnerable narcissism displayed moderate correlations with a In the interpersonal circumplex, grandiose narcissism was mainly related to interpersonal problems related to domineering (PA) and also vindictive (BC) interpersonal styles. Both were related more to the antagonistic than the agentic aspects. Vulnerable narcissism was associated with a wide array of interpersonal problems , most strongly those related to a socially avoidant interpersonal style. This was attributable to neurotic rather than antagonistic aspects of narcissism. Relations with interpersonal problems were generally highly similar for the different FFNI versions.Both grandiose and vulnerable narcissism were related to aggression, and, as expected, the associations were most pronounced for antagonistic narcissism within the three-factor model . Grandiose narcissism was weakly related to attachment avoidance, whereas vulnerable narcissism was substantially related to both, attachment anxiety and avoidance. Within the three-factor model, it was mostly neurotic, but also antagonistic narcissism, which related to both dimensions. Finally, grandiose narcissism was related to game-playing (ludus) and also pragmatic (pragma) love styles, and the same pattern was evident for its constituent three-factor dimensions agentic and antagonistic narcissism. Vulnerable narcissism and its neurotic aspects were further associated with an obsessive love style (mania).As expected, grandiose narcissism was moderately positively associated with self-esteem and life satisfaction , whereas vulnerable narcissism was negatively associated with both. Correlations within the three-factor model show that positive associations with adaptive adjustment are due to agentic narcissism, whereas negative associations are due primarily due to neurotic, but also antagonistic narcissism.Grandiose narcissism largely displayed zero-associations with psychological maladjustment (though the LF was negatively related to depression), whereas vulnerable narcissism showed strong positive associations with symptom load and depression. Within the three-factor model, agentic aspects of narcissism were not associated with symptom load and negatively associated with depression , antagonistic aspects were slightly positively associated with both, and neurotic aspects displayed strong and consistent associations.shame, indifference [reversed], and need for admiration), as well the antagonistic narcissism facet distrust. They had lower scores on the antagonistic narcissism facet lack of empathy. This pattern of results was the same for all questionnaire versions.To investigate the validity of the German FFNI regarding more objective clinical and forensic criteria, we studied differences relating to prior/current mental disorder diagnoses  and status as incarcerated young criminal offender. authoritativeness facet by trend only in the SF), which point in the direction of a highly grandiose-antagonistic narcissism profile, with low scores on neurotic aspects (need for admiration and shame).Also as expected, the sample of young offenders had significantly higher scores on grandiose narcissism and its constituent factor antagonistic narcissism, as well as significantly lower scores on neurotic narcissism in all questionnaire versions. Lower scores on overall vulnerable narcissism were evident only in the SSF and the Brief From (presented in the following). We observed significant differences on nearly all facets , which may impede its use in certain contexts, but also displays problems in its structure, in the way that the item set might not be optimally suited to capture the facet and factor structure in an unidimensional manner. Second, available short forms—the 60-item FFNI-SF and the 15-item FFNI-SSF—have been constructed selecting items based on high Item Response Theory discrimination parameters SF;  or princWe used the item selection procedure ACO to construct a 30-item brief form of the FFNI which preserves its structure and maximizes model fit, reliability, and validity of the three factors. We then compare the different FFNI versions regarding their internal structure and validity .N = 1,823, see N = 1,098, see N = 2,921) for further tests of the factorial structure in which we also evaluated the models described in Study 1, and examined criterion validity.We used the samples and measures described in Study 1 for the ACO item selection, cross-validation of the solution, and further factorial structure and criterion validity tests of the brief form. Specifically, for the item selection process (see below), we used data from those samples in which the FFNI-LF was administered .Based on this model, we optimized (1) the model fit, more specifically RMSEA, CFI, and SRMR, (2) the reliability  of the three factors, and (3) their correlations with external criteria. Regarding the latter, we maximized the correlations between the FFNI factors of agentic narcissism with NPI leadership/authority as well as NARQ admiration, antagonistic narcissism with NPI entitlement/exhibitionism, NARQ rivalry, and the PES, and neurotic narcissism with the HSNS and PNI vulnerable dimension. to ensure that pairs of semantically similar items were not selected to increase the internal consistency of the scale at the cost of construct coverage. We further excluded two reverse-coded items from the initial pool, as those would otherwise have been the only reverse-coded items in the pool, leading to a 52-item selection pool.As item selection pool, we used the FFNI-SF . We chosN = 1,823). To evaluate the robustness of the model fit, reliability and correlations of the scale, we then estimated these values for the final brief form on the combined sample in which participants filled in the short form . Because these samples had been collected independently from each other in the context of different studies, this approach should yield a good indication of the generalizability of the selected scale properties. Finally, for consistency with the analyses presented in Study 1, we re-evaluated the factorial structure in the overall pooled sample within the models presented there.We used ACO with a comparison of 50 models per iteration  and a stopping criterion of 30 iterations without improvement to the previously best solution. Because ACO is a probabilistic algorithm, a single run may not be sufficient to find an optimal solution. We thus ran it 10 times with different random generator seeds and used the best solution of the ten runs as final solution. To ensure that the solution was robust and generalizable across samples, we used cross-validation. Specifically, we selected the brief form solely in the combined sample in which participants filled in the long form . The BF showed largely satisfactory CFA model fit on the selection sample , as well as in the validation sample , and also in the pooled sample (see next section). Factor saturation omega was on average ω = .76 in the selection sample , and ω = .77 for all three factors in the validation sample. The average correlation between the three factors and the related scales used in the optimization was comparable across the selection and validation scale . In summary, the 30-item brief form yielded satisfactory psychometric properties, which were robust across different samples, and was highly correlated with the 60-item version, thus maintaining construct coverage.The 30 items selected by ACO for the Brief Form can be found in The 30-item FFNI-BF displayed good internal consistencies ranging from .72 ≤ α ≤ .84 at factor-level (two- and three-factor models) and from .60 ≤ α ≤ .88 at facet level. Internal consistencies were generally in between those of the FFNI-SF and FFNI-SSF, as it can be expected given the different scale lengths. The pattern of intercorrelations among the factors of the two- and three-factor models was generally highly similar to those of other scale versions see . as described in Study 1 in the overall pooled sample. As Supplemental Table S1 displays loadings of the FFNI-BF .To re-evaluate the factor structure of the FFNI-BF within the hierarchical CFA, ESEM, and ESEM-within-CFA framework, we tested the same modelsr = .26) while being weak in the FFNI-SSF (r = .11), and similar differences were evident for the entitlement/exploitativeness factor of the NPI, the entitlement rage facet of the PNI, and more generally the disagreeableness/antagonism measures (NEO-PI-R and PID-5). This is likely a result of including the correlation with entitlement—a core characteristic of both forms of narcissism , which is only weakly the case in the novel BF (r = .12) or the recently developed SSF (r = .14). Similarly, agentic narcissism is less strongly related to authentic pride in the shorter versions (r = .43 vs. r = .22 / r = .23), and less strongly related to self-esteem (r = .29 vs. r = .12 / r = .14) as well as life satisfaction (r = .28 vs. r = .08 / r = .08). Taken together, this shows that shorter scales draw less strongly upon adaptive aspects of extraversion, which potentially make them more suitable measures of agentic-extraverted narcissism, rather than extraversion per se .Correlations of the FFNI-BF with other personality measures and further external validity criteria were generally highly similar to those of the other scale versions reported in Study 1, with some notable exceptions: while agentic narcissism correlates most strongly with aspects of extraversion related to social dominance (NEO-PI-R assertiveness), the longer versions also display a substantial correlations with aspects related to positive emotionality  and lower scores on vulnerable narcissism (note that this was not the case in longer versions) and its constituent factor neurotic narcissism . At facet level, incarceration status was related to higher scores in the facets reactive anger (though only by trend), indifference, grandiose fantasies, manipulativeness (by trend), exploitativeness, entitlement, lack of empathy, arrogance, thrill seeking, and distrust, and lower scores in shame and need for admiration. The trend-level difference observed for authoritativeness in Study 1 was not evident in the BF. Taken together, the FFNI-BF is well suited to capture different characteristic FFNI profiles (see Study 1) at factor and facet level.Finally, the novel BF  displayed largely the same pattern of mean differences between groups with and without mental disorders diagnoses or incarceration status see . DiagnosStudy 2 presented the construction and validation of a 30-item brief form of the FFNI using the metaheuristic ACO algorithm. Results show that the FFNI-BF is a reliable and valid measure with high correlations to the FFNI-SF , an optimized internal structure (based on non-content-redundant items) resulting in superior fit in confirmatory analyses (without the use of ESEM), and high validity with respect to a range of relevant self-report variables and more objective clinical and forensic criteria. Specifically, we found highly similar associations with external criteria as for longer scale versions using a smaller number of items, making the FFNI-BF a highly efficient measure of narcissism within both the two- and three-factor model. One particular advantage compared to the recently developed SSF might be that the vulnerable narcissism factor in the BF does capture entitlement, which is regarded as a core characteristic of narcissism. Also, the FFNI-BF allows for facet-level scoring and yields similar facet-level differences with respect to clinical and forensic criteria as does the FFNI-SF. Taken together, the FFNI-BF may be recommended as a concise measure of narcissism when scores at the two- or three-factor level are needed, and may be interpreted also at the facet level in group-level statistics.The main aims of the studies presented here were to validate a German version of the FFNI—a measure covering the spectrum of narcissism from grandiose to vulnerable including agentic, antagonistic, and neurotic aspects from an FFM perspective—and to construct a brief form optimizing its internal structure and external validity. Overall, the data provide robust evidence for the validity of the German FFNI, including the newly devised 30-item FFNI-BF.The two and three-factor model scores of the German FFNI displayed high internal consistencies across all scale versions . At facet level, coefficients were between .70 ≤ α ≤ .89 for the LF and SF, and .60 ≤ α ≤ .88 in the new BF. These are similar to the original English scales .Confirmatory factor analyses showed generally acceptable model fit for most FFNI versions when an adequate level of complexity was taken into account: fit was better for hierarchical than nonhierarchical models, which shows that considering uniqueness at item and facet level is necessary when aligning FFNI data with the intended factor structure. When modeled only at facet level, fit was comparable to a previous study involving a confirmatory test of the FFNI-SF . Model fRegarding the question whether two- or three-factor models of the FFNI should be preferred, our data show that both are generally valid, though the three-factor solution showed slightly better fit in most cases see . These dr ≤ .74) and vulnerable narcissism . Estimates were similar to those obtained for the original English version (r ≤ .69), and FFNI antagonistic narcissism was primarily associated with antagonistic narcissism scales . Neurotic narcissism correlated most strongly with PNI vulnerability (.58 ≤ r ≤ .71), particularly its contingent self-esteem facet (.67 ≤ r ≤ .79). Generally, estimates were similar to previous studies (aggressiveness). FFNI neurotic narcissism might thus not relate to core narcissistic characteristics by itself, but rather reflect personality dysfunction , but not substantially with PID-5 detachment (likely due to its focus on extreme introversion). FFNI agentic narcissism also correlated with NEO-PI-R openness and conscientiousness (most notably achievement striving), and correlated negatively with agreeableness (most notably modesty). This pattern shows that the agentic narcissism factor does capture the agency / social dominance—aspect of extraversion, and is only weakly related to the aspect more characteristic of positive emotionality  and, interestingly, moderately associated with the PID-5 dimensions psychoticism, disinhibition, and detachment. FFNI neuroticism, as expected, correlated strongly with NEO-PI-R neuroticism, and with PID-5 negative affect. Beyond that, correlations with introversion/detachment were evident, and also with conscientiousness (competence and self-discipline) / disinhibition. As described above, FFNI neurotic narcissism was not directly related to scales assessing entitlement. It was even slightly positively associated with the NEO-PI-R facets modesty and compliance. Negative associations with facets of agreeableness were only evident for trust. FFNI neurotic narcissism was also not associated with PID-5 antagonism. Together, this points to FFNI neurotic narcissism assessing an emotionally instable but not entitled, socially anxious yet conforming personality. As The FFNI three-factor scores displayed the expected pattern of associations with nonclinical (NEO-PI-R) and clinical (PID-5) FFM measures: agentic narcissism correlated most strongly with NEO-PI-R extraversion  and with a more fine-grained measure of personality functioning (IPO). This is indicative of impairments in self- and other-related perceptual and regulatory capacities  which se stimuli . This alThe impairments associated with grandiose, particularly antagonistic narcissism reflect mainly in externalizing behavior, as we found that antagonistic narcissism was associated with domineering and vindictive interpersonal problems and self-reported aggression, including physical aggression. In the offender sample, antagonistic narcissism and almost all of its facets were significantly elevated, underlining the practical relevance of antagonistic traits with respect to criminal behavior , 2022.indifference and higher empathy (lower lack of empathy) might amplify mental health problems in these individuals in terms of over-involvement and empathic distress  correlated less strongly with NEO-PI-R extraversion facets related to positive emotionality , (2) shifted somewhat more toward the maladaptive pole in correlations with measures of psychological adjustment, and (3) correlated somewhat more strongly with PID-5 psychoticism, as well as aspects of the PNI. While the differences are subtle, they suggest that short versions of the FFNI agentic narcissism factor—constructed using different approaches each selecting a best-performing set of items—emphasize maladaptive aspects slightly more than longer versions. One advantage of this might be that the short scales seem to capture agentic-extraverted narcissism, not just extraversion per se, to a larger degree.A similar point applies to the vulnerable narcissism factor of the FFNI-BF, which—compared to the recently developed FFNI-SSF—shows a more substantial correlation with measures of entitlement (PES and NPI entitlement/exploitativeness) and disagreeableness/antagonism (NEO-PI-R and PID-5), similar to the longer scale versions. Since entitlement/antagonism are regarded as core characteristics of narcissism , the strWhile the research reported here provides promising evidence for the internal consistency, factor structure, and validity of the German FFNI and the novel FFNI-BF, there are limitations. First, validity evidence presented here relies mainly on self-report measures, which can be biased in the direction of more favorable self-presentations in grandiose narcissism cf. . Future We validated the FFNI in German-speaking samples and constructed a novel 30-item brief form. The FFNI generally displayed good internal consistency and convergent validity with other narcissism measures. Regarding its internal structure, we found that existing short versions align with the hypothesized factor structure when an adequate level of model complexity is taken into account. Guided by criterion-based optimization, we constructed a novel brief form which clearly depicts the factorial structure with a small item set, yet is more strongly saturated with theoretically relevant core characteristics. Regarding external validity, grandiose narcissism—particularly its agentic aspects—displayed correlations with validity measures indicative of egotistical but largely adaptive adjustment, but also impaired personality functioning. Vulnerable narcissism—particularly its neurotic but also its antagonistic aspects—was associated with maladaptive adjustment and markedly reduced personality functioning. Antagonistic and neurotic narcissism effectively discriminate between groups with and without a clinical and forensic history. We conclude that the German FFNI is a reliable and valid instrument for assessing narcissism from an FFM perspective, major strengths of which lie in its breadth spanning both grandiose and vulnerable aspects encompassing agentic, antagonistic, and neurotic narcissism, and the compatibility with general nonclinical and clinical personality models. The FFNI provides a comprehensive tool for assessing the distinct aspects of narcissism in a variety of research and applied contexts.Click here for additional data file.Supplemental material, sj-docx-1-asm-10.1177_10731911221075761 for Validation of the German Five-Factor Narcissism Inventory and Construction of a Brief Form Using Ant Colony Optimization by Emanuel Jauk, Gabriel Olaru, Eva Schürch, Mitja D. Back and Carolyn C. Morf in Assessment


Absolute values resources as constant across time and place while relative contextualizes one’s hierarchical location in the distribution of a time and place.A classic debate concerns whether absolute or relative income is more salient. This study investigates specifically whether absolute income or relative income matters more for health and well-being.We exploit within-person, within-age, and within-time variation with higher-quality income measures and multiple health and well-being outcomes in the United States. Using the Panel Study of Income Dynamics and the Cross-National Equivalent File, we estimate three-way fixed effects models of self-rated health, poor health, psychological distress, and life satisfaction.For all four outcomes, relative income has much larger standardized coefficients than absolute income. Robustly, the confidence intervals for relative income do not overlap with zero. By contrast, absolute income mostly has confidence intervals that overlap with zero, and its coefficient is occasionally signed in the wrong direction. A variety of robustness checks support these results.Relative income has far greater predictive validity than absolute income for self-reported health and well-being.Compared to earlier studies, this study provides a more rigorous comparison and test of the predictive validity of absolute and relative income that is uniquely conducted with data on the United States. This informs debates on income measurement, the sources of health and well-being, and inequalities generally. Plausibly, these results can guide any analysis that includes income in models. Income Through two key innovations, this study provides a unique test to adjudicate this debate. First we exploit within-person, within-age, and within-time variation within the United States specifically. Second we use higher-quality, state-of-the-art income measures, incorporating taxes and transfers and adjusting for household size. Incorporating these advances with multiple health and well-being outcomes, we provide a more rigorous comparison and test of the predictive validity of absolute and relative income that is uniquely conducted with data on the United States.2.The debate between absolute and relative income is critical for understanding inequalities and for normative debates. The debate also speaks to absolute and relative poverty measures, income interventions designed to improve health and well-being, and literatures on social class. As Absolute income measures resources independent of time and place. If a person or place gets richer and living standards rise, health and well-being should mechanically improve as absolute incomes increase . This imRelative income locates one relationally within a distribution of a time and place. Income is a positional good, and its value depends on one’s hierarchical rank compared to others in a given cultural and historical context . Income 3.Four kinds of studies predominate in this debate. First, laboratory-style experiments simulate absolute and relative resources and/or consumption . Second,While all these types of studies have been valuable, each has limitations. The first type is vulnerable because of the dubious external validity of laboratory experiments. Also, laboratory simulations of income or consumption are unlikely to be as construct valid as measures of actual economic resources. For the second type, it is unclear if subjective perceptions of relative deprivation in observational studies actually track objective material deprivation. While the third type has generated a voluminous literature, many question these ecological studies’ typically macro-level cross-sectional research designs, the overreliance on bivariate correlations, the vulnerability to omitted variable bias, and the non-robustness of results .The fourth approach avoids some of these problems. However, even those analyses are unable to measure both absolute and relative income at the person level. The mean income of a place or reference group confounds two very different concepts. On one hand, it captures the prevailing standards of a place or reference group – and a negative coefficient therefore tracks relative deprivation. On the other hand, the mean of a place or group includes a wide variety of unobserved characteristics of places or groups. In particular, the mean of a place or group includes the individual characteristics driving selection into places or groups.To be concrete about the present study’s contribution, we emphasize two innovations. In the process, we contrast our study with the relatively few similar prior panel analyses.within individuals. One advantage of within-individual analyses is that one can net out stable unobserved between-person characteristics, many of which could confound the relationship between income and well-being. A second advantage is that absolute and relative income are highly correlated in cross-sectional data, while there is far greater differentiation over time and when they are differenced from within-person means , which is ideal because of its large, nationally representative sample and multiple health outcomes observed at numerous time points . Further5Our analytic sample includes all observations with valid health and income measures through the 2019 wave. The number of valid cases differs by outcome. Across outcomes, the main analyses include 12,361–17,094 individuals for 60,317–198,396 person-years across waves  from 1984 to 2019.Our main analyses include all adult respondents. This includes “heads” with self-reports and spouses with proxy reports. We analyze heads only in 4.2The variable coding and descriptive statistics are shown in self-rated health  and poor self-rated health  . Life satisfaction is measured using a single item on a five-point scale: “Please think about your life as a whole. How satisfied are you with it?”   . We alsond poor)  for psyctisfied) .Our key independent variables – relative and absolute income – use post-fisc equivalized income. These incorporate taxes, tax credits , cash transfers , and near cash transfers . They adjust for household size by dividing by the square root of household members. Thus our measures explicitly improve on the conventional income measures in this literature, such as gross (pre-fisc) non-equivalized income.absolute income, we measure post-fisc equivalized income in real inflation-adjusted 2021 dollars. Following prior studies, we convert absolute income to natural log. relative income, we measure post-fisc equivalized income in rank percentiles. These absolute and relative measures are similar to those in related studies of health and well-being  as fixed effects (FEs). Some models adjust for education and marital status as time-varying covariates . This converts absolute income into a within-time measure, which is not strictly consistent with the concept of “absolute” and becomes closer to “relative.” Hence including wave FEs changes the interpretation of absolute income. Fortunately, the results are consistent regardless of whether we include wave FEs.The use of three-way FEs does require special attention, however. Therefore we estimate all models with and without the wave FEs (βWt). Person FEs are perfectly collinear with cohort FEs, so the classic age-period-cohort problem is triggered when person, age, and wave FEs are all included. We reduce this problem because the five-year age FEs differ from yearly age FEs. In addition, with wave FEs, absolute income is demeaned from each year’s mean absolute income  would be dropped from the poor health models because their outcome does not change. We strongly prefer to retain as many individuals as possible, including those with stable health and well-being. Indeed, if relative income or absolute income changes but health or well-being does not, this is certainly relevant. Second, logit models with FEs are vulnerable to the incidental parameters problem, which could result in bias  increase in relative income is associated with 0.06 higher self-rated health and a −0.02 lower probability of poor health. By contrast, a one SD increase in absolute income is associated with only 0.01 lower self-rated health and a 0.004 higher probability of poor health. Hence relative income’s x-stdBs are about 6.2 and 5.3 times larger than those of absolute income for self-rated health and poor health.For psychological distress, the x-stdB for relative income (−0.23) is far larger (46x) than the x-stdB for absolute income (−0.005). For life satisfaction, the x-stdB for relative income (0.05) is far greater (53x) than the absolute income x-stdB (−0.001).The largest x-stdBs for relative income are for psychological distress. A one SD increase in relative income is associated with 0.23 SD lower psychological distress. The next largest stdBs are for self-rated health (0.06), life satisfaction (0.05), and poor health (−0.02).th to the 75th percentile corresponds to roughly a 1.76 SD change in relative income. If relative income moved from the 25th to the 75th percentile, self-rated health would be expected to increase by 0.10 (on mean of 3.5; see For concrete substantive interpretation, note that a one SD change in relative income corresponds to 28.4 percentage points. Therefore moving from the 255.2Finally, 6.A classic unresolved debate concerns whether absolute income or relative income is more consequential for health and well-being. While absolute income is constant across time and place, relative income is contextualized as a hierarchical location within the distribution of time and place. This debate is central to inequalities and deprivation, norms about distributive justice, poverty measurement, and income-based interventions for health. It also should inform how scholars measure income in their analyses.Most of the literature uses cross-sectional data, and comparatively few studies examine variation within individuals over time. Prior longitudinal analyses have used weaker income measures. To the best of our knowledge, there has been only one unpublished panel analysis within the United States. By contrast, we exploit higher-quality income measures and multiple measures of health and well-being in the PSID-CNEF. Our three-way FE approach scrutinizes within-person/within-age/within-time variation and better differentiates between absolute and relative income.p-values than absolute income. Indeed, the comparative magnitude of the x-stdBs overwhelmingly suggests that relative income is more important than absolute income. Our results are also robust to a wide variety of alternative model specifications and samples.Our results demonstrate that relative income is more important than absolute income to health and well-being. For all four outcomes, relative income has much larger x-stdBs and far smaller Because our three-way FE approach nets out stable between-individual, between–time period, and between-age differences in health, our study contributes further convincing evidence of long-standing claims that income is a fundamental cause of better health . UniquelOur study has limitations that future research should address. We suggest five issues. First, the PSID added a greater variety of health outcomes only recently. As more time points become available for outcomes like chronic conditions and acute events, our approach should be replicated. Second, we split the analyses only by sex and age, but others can decompose our analyses further by other characteristics, such as race/ethnicity. Third, we examine absolute and relative income across the entire income distribution. It would be worthwhile to focus on lower-income populations to assess if our conclusions apply when distinguishing between relative and absolute poverty e.g., . Fourth,Fifth, we have advocated for our three-way FE approach. We have explained that within-individual analyses can net out stable unobserved between-person characteristics, many of which could confound the relationship between income and well-being. Also, because absolute and relative income are highly correlated in cross-sectional data, our approach enables clearer differentiation between the two. Despite what we view as clear merits, we acknowledge reasonable concern about whether within-person analyses can always generalize to between-person comparisons. Therefore there is a need for further research contrasting the well-being of people who vary in terms of absolute and relative income. The perennial challenges in clearly differentiating between absolute and relative income and unobserved confounding remain. Still, to augment the generalization that relative income better predicts well-being, between-person analyses are necessary.Despite these limitations and needs for further research, we propose that relative income should be centered over absolute income in health and well-being research and maybe even inequality research generally. Fortunately, it is feasible to calculate relative income in almost any dataset, as all it requires is to locate a case within the sample’s distribution. If cross-sectional data are used or it is not possible to include both relative and absolute income in the models, relative income should be prioritized. Relative income appears to be the more salient dimension of inequality and better captures the returns to affluence that higher incomes provide. Economic resources are positional goods, and income’s value for health and well-being is relational to and conditional on others’ resources.


To test the hypothesis that early-life adversity accelerates the pace of biological aging, we analyzed data from the Dutch Hunger Winter Families Study . DHWFS is a natural-experiment birth-cohort study of survivors of in-utero exposure to famine conditions caused by the German occupation of the Western Netherlands in Winter 1944–5, matched controls, and their siblings. We conducted DNA methylation analysis of blood samples collected when the survivors were aged 58 to quantify biological aging using the DunedinPACE, GrimAge, and PhenoAge epigenetic clocks. Famine survivors had faster DunedinPACE, as compared with controls. This effect was strongest among women. Results were similar for GrimAge, although effect-sizes were smaller. We observed no differences in PhenoAge between survivors and controls. Famine effects were not accounted for by blood-cell composition and were similar for individuals exposed early and later in gestation. Findings suggest in-utero undernutrition may accelerate biological aging in later life. Insults to early-life development are predicted by theory to impact trajectories of healthy aging –4. ConsiTo explore this alternative hypothesis, we analyzed blood DNA methylation (DNAm) data collected in a natural-experiment study of in-utero famine exposure to test differences in the pace and progress of biological aging between famine-survivors and matched controls. The Dutch Hunger Winter Families Study (DHWFS) enrolled a cohort of survivors of in-utero exposure to the Dutch Famine 1944–5), matched controls born before or after the famine in the same hospitals as the survivors, and their same-sex siblings . We comp44–5, matSI Appendix, Table S1).We analyzed data for N=951 cohort members with available DNAm data  and biological age (GrimAge and PhenoAge) between famine-survivors and controls. Second, we conducted dose-response analysis to test if participants who were exposed to the famine for more weeks of gestation exhibited larger famine effects as compared with those exposed for fewer weeks of gestation. Third, to explore specificity of famine effects to exposure during specific periods of gestation, we classified famine survivors according to when in gestation they were exposed, as described in the Methods section, and computed effect estimates for each window of exposure. In each step, we conducted analysis (a) in the full DHWFS; (b) using a between-families comparison of famine-survivors to unexposed time controls born immediately before or after the famine; and (c) using a within-family comparison of famine-survivors to their unexposed same-sex siblings. We also explored whether associations of famine exposure with biological aging differed between men and women.p=.018). Differences between famine survivors and controls were of smaller magnitude for GrimAge and PhenoAge and not statistically different from zero at the alpha=0.05 level . Results were similar in analysis restricted to include only famine survivors and unrelated time controls. Results are shown in SI Appendix, Table S2; full results for all biological aging measures are reported in SI Appendix, Table S3; a correlation matrix is shown in SI Appendix, Fig. S1.Cohort members exposed to famine in utero had faster pace of aging compared with unexposed cohort members . There were no dose response effects for GrimAge and PhenoAge . Results are reported in SI Appendix, Table S4; full results for all biological aging measures are reported in SI Appendix, Table S5.In dose-response analysis, cohort members who were exposed to the famine for more weeks of gestation had faster biological aging . Findings from sex-stratified dose-response analysis showed similar results . In sex-stratified analysis of gestational timing, results were different for women and men . For women, Dunedin PACE effect-sizes were similar across gestational time windows (effect-sizes ranged from β=0.07–0.25). In contrast, for men, Dunedin PACE effect-sizes were largest for later-gestational exposures and smaller for exposure in early gestation (effect-sizes ranged from β=−0.16–0.19). This pattern was similar for GrimAge. There was no consistent pattern for PhenoAge. Results are shown in We conducted exploratory analysis of sex differences in famine effects using sex-stratified regressions and analysis of effect-measure modification. In stratified analysis, famine effects were consistently larger for women and were near zero for men , famine survivors tended to be aging faster than their unexposed same-sex siblings; however, effect-sizes were attenuated by roughly half as compared with the full-sample analysis and were not statistically different from zero. In sex-stratified analysis, differences between sisters discordant for famine exposure (n=129 pairs) were nearly identical to famine-effect estimates from our original models whereas, among brothers (n=98 pairs), effect estimates were near zero or in the opposite direction of our original analysis. Results are reported in SI Appendix, Table S7–9).We repeated our analysis with additional covariates for estimated proportions of white blood cell types. In cell-count-adjusted analysis, effect-sizes were similar for DunedinPACE, GrimAge, and PhenoAge . We sampled an equal number of births in each month, allocated across the three institutions according to their size, to obtain 890 singleton births.same-sex sibling born before or after the famine would be available to participate. A total of N=547 of the famine-exposed group, N=176 of the time-control group, and N=308 same-sex unexposed siblings consented to participate and underwent a computer-assisted structured interview by telephone.Of the total famine-exposed and time-control births, current addresses were able to be traced for 70%. These individuals were invited by mail to join the study and were additionally asked if a Data Collection was conducted in 2003–2005, approximately six decades after the famine. Of the N=1,031 participants who were interviewed, N=971 also participated in a clinic exam . FollowiWe defined the period of famine from archival records of weekly ration distributions, as described previously . BrieflyFamine-exposed participants experienced an average of 17 weeks of gestation during which ration distributions were <900kcal/day. Following prior work with the cohort , 40, parBiological aging is the progressive loss of system integrity with advancing chronological age . BiologiSI Appendix, Supplementary Methods.Briefly, DNA methylation (DNAm) was measured from blood collected at the clinic exam using the Illumina Infinium Human Methylation 450k BeadChip and preprocessed as previously described . Furtherhttps://github.com/danbelsky/DunedinPACE). We calculated high-technical-reliability “PC” versions of the GrimAge and PhenoAge clocks developed by the Levine Lab (https://github.com/MorganLevineLab/PC-Clocks). The clocks are described in detail in the SI Appendix, Supplementary Methods.Our primary analysis focused on three epigenetic clocks for which validation data across multiple studies establish robust associations with healthspan and lifespan and sensitivity to exposures known to hasten aging-related health decline: the DunedinPACE clock, which measures pace of aging, and the GrimAge and PhenoAge clocks, which measure biological age. We calculated DunedinPACE using the R code available on GitHub . For comparison purposes, we report results in the d clocks , 48, 49.d clocks , 50, 51.d clocks  and PythThe analysis sample for this study was formed from participants in the clinic examination who provided a blood sample from which DNA was extracted and stored at LUMC. For our analysis, DNA were available for N=960 individuals. After quality controls, DNA methylation datasets were available for N=951. These individuals formed our analysis sample.We used regression analysis to test associations between in-utero famine exposure and DNAm measures of biological aging. First- and second-generation epigenetic clock values have high correlations with chronological age. For analysis and interpretation, the standard approach is to regress clock values on participants’ chronological age values and predict residual values. These values, often referred to as “age acceleration residuals”, aim to quantify the difference between how much aging a person has actually experienced relative to the expectation based on their chronological age. No residualization was performed for DunedinPACE, which is a rate measure and shows only moderate correlation with chronological age. To account for the non-independence of measurements taken from siblings, we used generalized estimating equation (GEE) regressions . Our mod


Rats administered microwave-water have shown a significant down-regulation in GST, Metallothionein and CYP genes’ expression, however, Amylase and HDAC3 genes were significantly up-regulated, compared to the other groups. The intake of microwave-water resulted in notable histopathological changes in liver tissues. Rats administered magnetic-water showed no clear changes in their liver tissues. In summary, microwave-water induced stress and epigenetic effects compared with magnetic-water and tap-water. Also, magnetic-water produced from the higher magnetic power had no side effect on liver tissues.Physical and chemical changes in the natural of water may affect biological organisms. In this study, we highlight the effect of magnetized-water and microwave-water on rats’ liver tissues. Three groups of albino rats were separated. The first, rats were administered tap-water. The second, rats were administered magnetized-water. The third, rats were administered microwave-water. After two months, the results revealed a significant increase in liver functioning enzymes’ levels and bilirubin in rats administered microwave-water, compared to tap- and magnetic-water. In relation to oxidative stress, there was a significant increase and decrease in oxidative and antioxidant parameters respectively in liver tissues of rat's administrated microwave-water. At the molecular level, there was a significant down-regulation  Due to its crucial vital rule to most living organisms, numerous physical and chemical treatments have been developed to enhance drinking water quality2. Magnetic-water is created by passing tap-water through magnetic field (magnetic tubes). As a consequence, water properties become extremely fertile and active; resulting in an increasement of oxygen ratio, velocity of salts and amino acid dissolution3. Unlike dead water, the pH of magnetic-water changes from acidity to alkalinity. Also, magnetic-water’s weight and salinity increase4. Magnetic-water improves the circulation of blood and oxygen, transportation of nutrients in blood, activation of enzymes, elimination of internal toxins resulted from metabolism, absorption of internal toxins and improves the blood picture and sexual hormones3.Water is the most abundant substance on planet Earth. It is the primary component of most living organisms. Approximately, 65% of human body weight consists of water. In all practical respects, water is the only solvent in human body5. Millions of people worldwide rely on microwave ovens on daily basis for food cooking and drinks preparation.It has been reported that microwave ovens usage has led to the formation of cancer-causing substances due to; (a) releasement of toxic chemicals from food packaging into food; (b) the destruction of nutrients; (c) decreased bioavailability of vitamins , essential minerals and lipotropic factors in food; (d) loss of natural food antioxidants. Symptoms of direct exposure to microwave ovens radiation include nervous disorders such as dizziness, headache, irritability, sleeplessness, nervous tension, anxiety, and inability to concentrate6. The FDA advises consumers to avoid the risk induced by boiling water in the microwave oven.7The most important polar molecule in foods is water. Water molecules change direction with the effect of microwavesThis study aims to evaluate the effect of water treated with physical activity, such as magnetic and microwave apparatuses, on rats’ liver tissues at the molecular, epigenetic, histological levels and oxidative stress. The novelty of this manuscript lies in factoring magnetic and microwave treatments on the level of stress and epigenetic effects on liver tissues of rats.8. According to the current study, liver enzymes levels  did not significantly change in rats administered tap-water as well as rats administered magnetic-water , globulin ratio, antioxidant status, water quality, semen quality, blood picture, liver, and kidney functions. Consequently, improved buck fertility. Regarding water treated with microwave, our results may agree with Zhang14. They suggested that the non-ionizing electromagnetic radiation effect (microwave oven) is linked to an increased risk of cancer, particularly in residential exposed populations within the United States. Additionally, genetic material in body tissues of those exposed to non-ionizing electromagnetic radiation from sources such as microwave ovens will change over time8. According to reports, microwave oven usage can result in the formation of substances that cause cancer, release of toxic chemicals from food packaging and destruction of nutrients5. Vitamins B complex, C, and E reduced bioavailability, as well as essential minerals and lipotropic factors in food and loss of antioxidants5. Microwave water treatment deleteriously impacted Japanese quails’ development and numerous physiological traits5. In our investigation, protein fractions' findings revealed little variations in plasma concentrations of total protein, albumin, and globulin in rats administered tap- magnetic- and microwave-water, separately.Recent evidence confirmed that magnetic-water has an advantageous impact on growth parameter. Tissues of the human body are altered by exposure to non-ionized electromagnetic radiation, such as that from microwave ovenser Table . However13. The authors found that magnetized water significantly increased antioxidant enzymes and decreased lipid peroxidation biomarkers in rabbit bucks. This might be due to applying 4000 G, whereas in our study we applied higher magnetic power (14500 G) for water treatment. This led us to conclude that utilizing high magnetic power reduced the positive effect . When rabbit bucks were administered water treated with 3600 G for 90 days, blood markers of oxidative stress  decreased, while total antioxidant capacity and antibody titer increased15. According to several studies, magnetized water can have a significant impact on the oxidant-antioxidant balance such as reducing Malondialdehyde and nitric-oxide levels, boosting superoxide dismutase enzyme activity in the heart, kidney and liver; all of which reduce the oxidative stress16.Our results in Table  revealedGST, Metallothionein, CYP, Amylase and HDAC3 genes were investigated. Authors of several articles investigated the increase, and decrease, in genes expression in response to exposure to magnetic field, not magnetic water. In Shewanella oneidensis, a magnetic field of 14,100 G accelerated the transcription of 21 genes, while slowing the rate of transcription in 44 other genes17. The accelerated genes include unknown-function genes (seven genes) and known-function genes. Two genes related to the transport and binding proteins, Methyl-accepting chemotaxis protein for cellular process function, UmuD protein for DNA metabolism, and OmpA family protein for cell envelope, were accelerated. The regulatory function includes transcriptional regulator, LuxR, LysR, and MarR family. For transcription function, there are two genes; pnp and Polyribonucleotide nucleotidyltransferase. In addition, deoC and Deoxyribose-phosphate aldolase were accelerated. There are two genes related to amino acid biosynthesis; Homoserine kinase (thrB) and, Diaminopimelate decarboxylase(lysA). Fatty acid and phospholipid metabolism function include Acyl-CoA dehydrogenas family; Other genes include Iron-containing alcohol dehydrogenase were accelerated. On the other hand, downregulated genes include five genes related to energy metabolism  and all genes related to Amino acid biosynthesis . Bacterioferritin subunit 1 gene (bfr1) related to transport and binding proteins function downregulated. Also, for the cellular process; fic and mine, and DNA metabolism function; Type I restriction-modification system and M subunit downregulated. Three cell envelope genes include chain length determinant protein, Polysaccharide biosynthesis protein and pile dowenregulated. For regulatory function; DNA-binding response regulator and Transcriptional regulator LysR family downregulated. In addition, there was an ATP-dependent RNA helicase and DEAD box family for downregulated transcription genes. For Purines, pyrimidines, nucleosides, and nucleotides there were carB and hpt-2 genes. Amino acid biosynthesis includes asnB gene. Peptidase, putative, and Heme exporter protein CcmD are genes related to protein fate functions that are downregulated. Other genes downregulated include Glyoxalase family protein, OmpA-like transmembrane domain protein, TPR domain protein and ISSod3, transposase.In order to confirm the effect of magnetic- and microwave-water at the molecular level, expression profiles from c-myc gene in mice and humans18. It was reported that magnetic field interferes with normal genome function at the level of gene expression (magnetic field-induced expression)19. In our study, magnetic-water significantly reduced gene expression of Metallothionein “Fig. 20. The authors found that female rats' plasma Metallothionein activities were boosted in response to short-term exposure to a magnetic field; and suggested that such increases in Metallothionein activities indicating that biosynthesis of Metallothionein might have been induced by stress20. The fact that magnetic-water, in our results, did not induce the stress that was induced in presence of direct magnetic-field agrees with Chater et al.20. Previous findings indicated that magnetic-water had no negative effect on liver’s health. We can clearly see that the indirect exposure to magnetic-field (magnetic-water) was not harmful. Also, down-regulation of CYT gene disagrees with the increased levels of CYT in the study reported by Patruno21 in response to magnetic-field, not magnetic-water in our study as discussed earlier. This is the first time to study magnetic-water and its effect on the expression of genes investigated. In contrast, there was no significant expression differences, between magnetic-water and tap-water, in the remainder of the investigated genes . When biological material is exposed to magnetic radiation, it absorbs a certain amount of energy, which depends on the dielectric properties of the biological material22. This non-ionizing electromagnetic radiation is absorbed, at the molecular level, and shows up as changes in the molecules’ vibrational energy or heat. Microwaves cause different biological effects; through adding ions in different locations. As a consequence, the speed and/or direction of biochemical reactions is modified23. Cell treatment with and without water is different as described by Szopińska and Dorna24. In addition, the different results show that water "the different factor in two experiment" had an obvious effect on the living cell. Significant down-regulation of GST, Metallothionein and CYP genes and up-regulation of Amylase and HDAC3 genes induced by microwave-water, compared to tap-water, indicated the deleterious effects of microwave treatment. These effects could be due to the changes in the water properties such as pH, conductivity and mobility of water molecules25. We also have noticed the precipitation of salt after boiling water in the microwave oven; this precipitation due to attaining supercritical condition26 and it could be the main reason that causes the negative effects of microwave treatment. Along with other changes in the normality of water that were mentioned by other authors led to vital changes induced in living cells. The fact that water is highly dipolar is what makes it able to dissolve salts. This makes it possible for water molecules to form spheres around the separated salt ions. As the temperature goes up, structure of the solution (hydrogen-bonding network and the clustering of water molecules around solutes) slowly changes. This is caused by an increase in the molecules’ kinetic energy and a corresponding decrease in their molecular interaction. The effective electrostatic force, associated with ion charges, diminishes with temperature at a much slower pace than the water dipole moment does. As a result, the hydrogen-bonded network weakens and cannot provide adequate protection from the salt ions26. These alterations in tap-water properties could lead to epigenetic effects on the genes. This is very clear in the highly induced regulation of HDAC3 gene by microwave-water. HDAC3 belongs to genes that play a major role in epigenetic effects. HDAC3 encodes for a protein that belongs to histone deacetylase. Microwave-water induced up-regulation in this gene, compared to tap- and magnetic-water. Therefore, we can conclude that microwave-water had a role in epigenetic effect.Weak magnetic fields, ranging from 0.8 to 8.0 G, promoted the transcription of the 27; they reported negative effects on liver tissues of Cyprinus carpio fish. Studies on the histological effects of magnetized water on animals, especially rat liver, appear to be scarce. However, Hassan et al.28 reported that prolonged exposure to magnetized water at 0.2 Tesla may result in liver damage of Scortum barcoo fish; based on histopathological observations of the liver. Aboulfotoh et al.29 demonstrated the impact of water (treated with either a magnetic field or microwave oven) on bone minerals and concluded that magnetized water has a beneficial impact on bone minerals. However, they also reported that microwave-treated water had an unfavorable effect on bone minerals; leading to decreased minerals than those reported in the control and magnetic water groups.Histological examination of liver tissues in rats administered tap-water revealed the presence of normal hepatocytes and dilatation in the central vein Fig. A. SimilaIn the last, we can conclude that some physical, not all, can change the nature of water and consequently affect the organism. This study concludes that microwave-treated water changed the liver function enzymes, oxidative stress, and antioxidant parameters. In addition, it changed the gene expression for important genes. Magnetically treated water had no side effects compared with normal water.Sprague Dawley albino male rats  were housed in cages under standard conditions . Rats were fed on pelleted diet containing the following nutritional ingredients: soybean meal (11%), sunflower oil (15%), yellow corn (49%), concentrate mixture (10%), wheat bran (10%), common salt (0.5%), molasses (3%), dicalcium phosphate (0.1%), ground limestone (0.2%), dl-methionine (0.7%), lysine (0.2%) and mineral-vitamin premix (0.3%)30. Rats had free access to water and food. Upon rats’ adaptation, they were randomly distributed into three groups; each group contained seven rats. Rats of the first group (negative control) administered natural tap-water for the entire duration of the experiment (two months). The second group of rats (magnetic-water group) administered magnetic-water produced from natural drinking water by passing through a 14,500 Gauss (G) magnetic field within a device (magnetizer) that was purchased from Delta Water Company .  Third group of rats (microwave-water group) administered tap-water treated with microwave oven  for 10 min in a glass beaker until boiling and then cooled at room temperature29 (the time was approximately ten minutes from put the water into the oven till boiling).This study was conducted at the animal house of Delta University of Science and Technology. The experimental design was approved by Delta University's ethical committee. Twenty-one By the end of the experiment timeline (second month after adaptation), rats went through overnight fasting process. Rats were then euthanized by exsanguination. Blood samples were collected from the portal veins of the liver. Sera were separated by centrifugation at 4000 rpm for 10 min. The liver was immediately removed, instantly washed with isotonic saline, and weighed. Then, liver was split into three parts; (a) first part was kept in − 80 °C for further RNA and molecular analysis; (b) second part was fixed for further histopathological examination in formalin 10%; (c) third part was homogenized for further antioxidant analysis.31.Alkaline phosphatase (ALP), aspartate transaminase (AST) and alanine transaminase (ALT) activities were measured using kits from Diamond-Diagnostics . Kits from BioMed-Diagnostics were utilized to measure gamma-glutamyl transpeptidase (GTP), albumin and bilirubin 32. NO, Superoxide dismutase (SOD) and Catalase (CAT) were estimated according to Archer33, Paoletti,34 and Aebi35, respectively.Antioxidants and oxidative markers activity measurement took place on homogenized tissues from liver. One gram of liver tissues was homogenized in 9 ml ice-cold phosphate-buffered saline, followed by centrifugation at 10,000 rpm under cooling (4 °C). Estimating various antioxidants levels took place on the supernatant via colorimetric and ELISA methodology, using Rayto ELISA microplate reader . Lipid peroxidation via thiobarbituric acid-reactive substances (TBARS) methodology36) utilizing sequences published from the rat genome project (Table GAPDH)37.Total RNA from samples representing each individual in each group was extracted, using Direct-zol™ RNA MiniPrep Plus kit  in three technical replicates. Extracted RNA quality and concentrations were evaluated using NanoDrop™ (ND-1000) spectrophotometer . The SuperScript™ IV One-Step RT-PCR kit  was used to generate cDNA from extracted RNA; followed by diluting all cDNA samples to the same concentrations using NanoDrop™ (ND-1000) spectrophotometer. Real-time qPCR reactions included cDNA as a template, QuantiTect™ SYBR Green qPCR Master Mix and gene-specific primers. Primers were designed using web-tool Primer3web™  and Duncan tests were utilized to conduct statistical analysis . Statistical results were reported as the mean ± standard error (SE). Differences between means are deemed statistically significant at The experiment was carried out under Egyptian ethical codes for studies on experimental animals and approved by the Animal Ethical Committee, Delta University’s ethical committee, Egypt  and all experiments were performed by relevant guidelines and regulations. The study was carried out in compliance with the ARRIVE guidelines.


The Kyrgyz Republic is a high-burden country for rifampicin resistant/multi-drug resistant tuberculosis (RR/MDR-TB). TB control efforts rely on early diagnosis and initiation of people on effective regimens. We studied the interval from diagnosis of RR-TB to starting treatment and risk factors for unsuccessful outcomes among people who started RR/MDR-TB treatment in 2021. We conducted a cohort study using country-wide programme data and used binomial regression to determine associations between unsuccessful outcomes and predictor variables. Of the 535 people included in the study, three-quarters were in the age category 18–59 years, and 68% had past history of TB. The median (IQR) time from onset of TB symptoms to diagnosis was 30 (11–62) days, 1 (0–4) days from diagnosis to starting treatment, and 35 (24–65) days from starting treatment to receipt of second-line drug susceptibility test (SL-DST) results. Overall, 136 (25%) had unsuccessful outcomes. Risk factors for unsuccessful outcomes were being homeless, fluroquinolone resistance, having unknown HIV status, past TB treatment, male gender and being unemployed. Treatment outcomes and the interval from diagnosis to starting treatment were commendable. Further reductions in unsuccessful outcomes by be achieved through ensuring timely diagnosis and access to SL-DSTs and by reducing the proportion of people who are lost to follow-up. Tuberculosis (TB) is a global public health challenge and, until the coronavirus, was the leading cause of death from a single infectious disease (ranking above HIV/AIDS). An estimated 10.6 million people fell ill with TB globally in 2021, and 1.6 million (15.1%) of them died [The Kyrgyz Republic, with an estimated incidence of RR/MDR-TB of 49 per 100,000 population, is among the 30 countries with a high-burden of RR/MDR-TB . BiomediResistance to fluoroquinolones (FQs) is associated with unsuccessful outcomes among people on RR/MDR-TB treatment ,7. AccesTB has biomedical and socioeconomic dimensions. Hence, the role of social factors in influencing unsuccessful outcomes cannot be ignored. Studies have reported poorer outcomes among key populations  ,12,13. WCohort study using programme data.2 and a population of 6,747,300 people [The Kyrgyz Republic is a middle-income country with an area of 198,500 km0 people . The couThe proportion of RR/MDR-TB cases is 27% among new cases and 59% among previously treated TB cases . There aTB treatment is provided free of charge. Patients receiving treatment as outpatients have to be registered with the Family Medicine Center in order for them to benefit from a package of free medical services. They also have the right to apply to local authorities for material assistance.All people who are presumptive of TB are tested for TB using smear microscopy and Xpert MTB/RIF assay , as per national guidelines. Once resistance to rifampicin is detected, a patient is started on second-line treatment. At that time, a second sputum specimen is sent to the national reference laboratory for both culture and phenotypic DST on MGIT and Lowenstein Jensen (LJ) media, and genotypic first-line and second-line DST using Hain Line Probe Assay .All patients with RR-TB are immediately started on the SHORRT regimen 4–6 (Bdq-Mfx-Cfz-E-H-(HD)-Pto/5 (Mfx-Cfz-E-Pto) pending DST results. Treatment regimen is decided on a case-by-case basis by DR-TB Consilia. The Consilia determine the composition of regimens, including number of drugs that are considered effective to maximize treatment success, reduce treatment failure and adverse drug events (ADEs). During the study period, there were no reports of stock outs of medicines to treat RR-TB. Patients in whom FQ resistance is presumed are started on a bedaquiline, pretomanid and linezolid (BPaL) regimen and, upon receiving second-line DST results, may be switched to individual regimens as shown in Patients are managed by either video directly observed therapy (VDOT), or family or facility DOT. They undergo regular clinical and laboratory tests to monitor both effectiveness of treatment and potential ADEs. Drugs that lead to serious ADEs are stopped and data on ADE are reported systematically using appropriate reporting forms (yellow forms).Patient level data are recorded in three interlinked electronic management systems: the Laboratory Management Information System, the electronic TB register and the electronic medical card. This makes it possible for requesting clinicians to access RR-TB results as soon as they are uploaded by laboratories. Treatment outcomes are documented as per WHO guidelines, as shown in Some patients in pilot sites who are on ambulatory treatment receive USD 12 monthly, which is transferred to their bank accounts. The money is meant to cover expenses on transport and nutrition, and is conditional on patients adhering to scheduled visits. The Red Cross Society provides monthly food packages to all vulnerable TB patients including migrants, ex-prisoners, HIV co-infected, low-income people and the homeless. People with TB are cushioned against transport costs to medical facilities using local funds.The study population is all patients who started second-line drug regimens for RR/MDR-TB and XDR treatment in Kyrgyz Republic from January to December 2021.The following variables were collected for the purposes of the study. Age in years; sex; migrant status; comorbidities ; alcohol use; smoking status; drug use status; homeless status; employment status; type of TB ; region ; place of residency ; whether sputum specimen was sent to reference laboratory (Yes/No); whether second-line DST was performed (Yes/No); fluoroquinolone resistance profiles; dates of (i) onset of TB symptoms, (ii) TB diagnosis, (iii) TB treatment initiation, (iv) receipt of second-line DST results; and outcomes of RR/MDR and XDR-TB treatment . Data sources were laboratory management information system (LMIS), the electronic TB register and electronic medical card.Patient-level data were entered into various electronic databases as a part of routine work. Each patient record had a unique identifier that was used to track the patient in the three databases. The censor date for ascertaining outcomes was 28 February 2023. Electronic data were exported from various databases to MS Excel. Once in Excel, data were cleaned to remove duplicates and fuzzy lookup was used to merge the three Excel databases into a single database comprising patient and laboratory data.p ≤ 0.05.Patient-level data were cleaned in MS Excel and were imported into Stata v13  for further cleaning and analysis. A new binary variable for treatment outcome (success or unsuccessful) was created. Categorical variables were summarised using frequencies and percentages. Continuous variables were assessed for normality using the Shapiro–Wilk test. Skewed data were presented as medians and interquartile ranges (IQRs). Time taken to receive second-line DST results were obtained by subtracting date of starting treatment from date of receiving DST results and were presented as medians and IQRs. We used the median cut-off of one day for time to start standardized second-line treatment to create a binary variable indicating either early or delayed treatment. Binomial regression was then used to test the association between unsuccessful outcomes and sociodemographic and clinical factors and results were presented as relative risks (RRs) and 95% confidence intervals. Level of significance was set at The sociodemographic characteristics of the 535 people with RR/MDR-TB who were enrolled in the study are presented in p = 0.99.Of the 496 people with valid dates, the median time from onset of TB symptoms to TB diagnosis was 30 days. Migrants and homeless people had median times (IQR) of 35 days (21–58) and 56 days (16–176), respectively. There was no difference in diagnostic delays by history of TB, Overall, 256 (55%) people started treatment within a day of RR/MDR-TB diagnosis, while 75% started treatment within 4 days of RR/MDR-TB diagnosis. Using the median turnaround time of one day as a cut-off, 55% of the people received treatment within a day. Half of the people received second-line DST results within 35 days, as shown in Treatment outcomes are shown in The factors associated with unsuccessful outcomes are presented in In this study we found longer time intervals from onset of TB symptoms to RR/MDR-TB diagnosis and from starting treatment to accessing second-line DST results, but shorter intervals between diagnosis of RR-TB and starting treatment. There were huge gaps in documenting HIV status. The proportion of unsuccessful outcomes was remarkably low and risk factors for unsuccessful outcomes were being homeless, FQ resistance, having unknown HIV status, past TB treatment, male gender and being unemployed.A previous study on prison inmates in the Kyrgyz Republic reported median delays from diagnosis to starting treatment of 7 days and further delays in people with RR-TB . In otheThere were marked delays among the homeless and migrants. Reasons for delays in being diagnosed with RR-TB (diagnostic delays) may be multifactorial. On the one hand, diagnostic delays may be due to person related factors such as delayed/poor health-seeking behaviour. People may delay health-seeking when they think the illness is due to other causes  or when FQ resistance is one of the risk factors for unsuccessful outcomes ,7. In thHIV is one of the risk factors for TB and TB/HIV co-infection is associated with poor treatment outcomes. The high proportion of undocumented HIV status is worrisome and has been reported previously . Though Several studies have documented treatment outcomes in the Kyrgyz Republic that range from 50% to 83%, depending on type of regimen . The outOur study is strengthened by the fact that we used nationwide programme data for all people who started on treatment in 2021. As a result, our findings reflect the situation on the ground and were free from selection bias.We acknowledge some limitations. Firstly, as with most programme data, we had some missing data. Secondly, we could not calculate delays in starting individualised treatment since dates of starting individualised treatment were not captured. Thirdly, the data we had made it difficult to tease out the people who were switched to individualised regimens. This made it difficult to disaggregate analysis by type of regimen. Shorter regimens are associated with successful outcomes as compared to standard of care ,5. ThirdDespite these limitations, our results have important considerations for both policy and practice. Firstly, delays in receiving diagnoses of RR-TB may be reduced by increasing awareness about TB symptoms within communities and the benefits of early health seeking for TB, the use of TB self-check apps and enhanced collaborations between the public sector and the private sector. Public–private partnerships may increase case notifications by tapping into people who first visit private facilities. Secondly, there is need to strengthen collaborations for TB/HIV and other comorbidities including diabetes mellitus. This ensures that those diagnosed with the comorbidities are linked to care to improve treatment outcomes. Both HIV and diabetes are risk factors for TB, and are responsible for worse outcomes as observed in this study and elsewhere. Thirdly, the long turnaround time for DSTs, including FQ, implies that people take suboptimal therapies for a long period before they are switched to effective and individualised therapies. From an operational point of view, there is need to improve efficiency of existing systems to shorten the time to accessing second-line DSTs. The TB programme may also consider investing in point-of-care rapid diagnostic technology such as the Gene Xpert MTB/XDR  that offers resistance profiles for isoniazid, rifampicin, FQs and second-line injectables ,33. ThisBoth treatment outcomes and the interval from diagnosis to starting treatment were commendable. Further reductions in unsuccessful outcomes in the Kyrgyz Republic are possible through ensuring timely diagnosis and access to SL-DSTs and by reducing the proportion of people who are not evaluated or lost to follow-up.


Single-cell chromatin accessibility sequencing (scCAS) technology provides an epigenomic perspective to characterize gene regulatory mechanisms at single-cell resolution. With an increasing number of computational methods proposed for analyzing scCAS data, a powerful simulation framework is desirable for evaluation and validation of these methods. However, existing simulators generate synthetic data by sampling reads from real data or mimicking existing cell states, which is inadequate to provide credible ground-truth labels for method evaluation.cis-regulatory interaction inference. We anticipate simCAS will be a reliable and flexible simulator for evaluating the ongoing computational methods applied on scCAS data.We present simCAS, an embedding-based simulator, for generating high-fidelity scCAS data from both cell- and peak-wise embeddings. We demonstrate simCAS outperforms existing simulators in resembling real data and show that simCAS can generate cells of different states with user-defined cell populations and differentiation trajectories. Additionally, simCAS can simulate data from different batches and encode user-specified interactions of chromatin regions in the synthetic data, which provides ground-truth labels more than cell states. We systematically demonstrate that simCAS facilitates the benchmarking of four core tasks in downstream analysis: cell clustering, trajectory inference, data integration, and https://github.com/Chen-Li-17/simCAS.simCAS is freely available at  Rapid advances in single-cell sequencing technologies have enabled the characterization of cellular heterogeneity and identification of disease-specific processes at the single-cell level . A rangecis-regulatory inference methods  low-dimensional embeddings generation, specifically in cell-wise and peak-wise; (ii) estimation for distributions of statistics, including the library sizes of the cells, the non-zero proportion of the cells, and the count summation of the peaks; (iii) parameter matrix correction, by leveraging information from the aforementioned estimated distributions of statistics; and (iv) synthetic peak-by-cell matrix generation, i.e. generating the final count matrix from a Poisson distribution. Note that a number of peak-by-cell count matrices are routinely binarized for downstream analyses, we thus provide an adapted framework with a Bernoulli assumption . Batch eThe cell- and peak-wise embeddings are first generated to represent the simulated cells and peaks, respectively. The properties of cells and peaks can be partly adjusted with different operations of generating embeddings. Given the number of simulated cells as The generation procedure of the homogeneous CEM For the heterogeneous CEM, we denote .,. is the element in where Given a real peak-by-cell count matrix, simCAS estimates distributions of three core statistics from real data: library size , cell non-zero proportion , and peak summation . The estimate distributions of these three statistics are used to correct the parameter matrix, which generates simulating count matrix, and a precise estimation improves the resemblance of synthetic data to real data.For the pseudo-cell-type mode, the distributions of three statistics are estimated from the input peak-by-cell matrix for each cell type. In this mode, similar to simATAC , the library size and the zero-peak proportion in each cell (cell sparsity) as with simATAC, and measure the diversity between the simulated data and real data by calculating median absolute deviation (MAD), mean absolute error (MAE), root mean square error (RMSE), Pearson correlation coefficient (PCC), Jensen–Shannon divergence (JSD), and Kolmogorov–Smirnov statistic (KSS). As a side note, for the library size, we perform log-transformation before comparison as with the recent benchmark studies , Homogeneity score (Homo), and adjusted Rand index (ARI) . In this section, simCAS was benchmarked against two baseline methods, simATAC and scMultiSim. Using the peak-by-cell matrix and the cell-type labels of each dataset as input, simCAS and baseline methods simulated the peak-by-cell matrix for each cell type with the same number of peaks and cells as in the real datasets.For statistical evaluation, we first performed comparisons of three properties, namely peak mean, library size, and cell sparsity of synthetic data to real by cell type (Section 2). For biological evaluation, we assessed the similarity between simulated cells and real cells for each dataset by calculating the miLISI value of mixed cells (Section 2). The results of data visualization and biological evaluation are shown in The cell types in most real scCAS data are obtained by unsupervised cell clustering and manual annotation, which highly rely on the investigator’s background knowledge, and may be not accurate enough as the ground truths for quantitative evaluation to analysis methods . Therefond σ=0.7 . We set  dataset . To furt dataset .K-means clustering, and HC  of discrete data with different number of cells (500–3000) and different number of populations (3–7) to evaluate three commonly used unsupervised clustering methods . First, we extended gene regions in Chromosome 1 upstream by 50 kb and downstream by 50 kb to obtain peak hubs. We then filtered the hubs containing <50 peaks and the hubs overlapped with others. Fourteen peak hubs (referred to as hub_1 to hub_14) are retained with the number of peaks ranging from 51 to 92 and the hub length ranging from 337 196 to 1 622 989 bps . FinallyF1 score for each pair of peak sets between predefined peak hubs and predicted CCANs, and evaluated the performance by the highest F1 score for each peak hub across all the CCANs. As shown in F1 score when setting the co-accessibility score cutoff to 0.5, which is reasonable because the possible interactive peaks will be discarded with higher cutoffs and more false positive samples may be predicted with lower cutoffs. With the penalization on the correlations by the distance, Cicero failed to annotate the regions of length significantly exceed the setting window, such as the hub_1 and hub_7 (cis-regulatory interaction inference.With the simulated data, we performed Cicero with the window parameter of 200 000 to predict the co-accessibility scores for peak–peak pairs in the simulated data, and the CCANs are constructed by the predicted interactions. Considering the interactive peaks as positive samples and non-interactive peaks as negative samples, we calculated a nd hub_7 . We furtWe developed a Python package simCAS, an embedding-based simulator of scCAS data, to simulate data from user-defined low-dimensional embeddings. With statistical evaluation and biological evaluation using multiple datasets with different protocols, and with different sizes, dimensions, and qualities, we illustrated that simCAS not only preserve cell- and peak-wise properties, but also capture biological signals. By testing various analysis methods on simulated data, we also demonstrated the capability of simCAS for benchmarking analysis, suggesting that simCAS has the potential to accelerate development of computational methods for scCAS data analysis. Besides, for the binarized input peak-by-cell matrix, the adapted simCAS framework with a Bernoulli assumption also showed the satisfactory capability to conduct benchmarking analysis .We also describe several avenues for improving simCAS. First, we can develop an R version of simCAS to offer users the convenience for benchmarking methods programmed in different languages. Second, we can focus on higher-level properties in real scCAS data, such as cell–cell correlation or variance at cell-wise and peak-wise. Third, we can incorporate external batch information from real data with data modeling in simCAS, to capture the characteristics of real batch effects. Fourth, as suggested by EpiAnno (btad453_Supplementary_DataClick here for additional data file.