Pan troglodytes verus. We examined whether emotional closeness, defined as attachment history with the yawning model, affected the strength of contagion, and compared the contagiousness of yawning to nose-wiping. Thirty-three orphaned chimpanzees observed an unfamiliar and familiar human (their surrogate human mother) yawn, gape and nose-wipe. Yawning, but not nose-wiping, was contagious for juvenile chimpanzees, while infants were immune to contagion. Like humans and dogs, chimpanzees are subject to a developmental trend in susceptibility to contagious yawning, and respond to heterospecific yawn stimuli. Emotional closeness with the model did not affect contagion. The familiarity-biased social modulatory effect on yawn contagion previously found among some adult primates, seem to only emerge later in development, or be limited to interactions with conspecifics. The influence of the ‘chameleon effect’, targeted vs. generalised empathy, perspective-taking and visual attention on contagious yawning is discussed.Contagious yawning has been reported for humans, dogs and several non-human primate species, and associated with empathy in humans and other primates. Still, the function, development and underlying mechanisms of contagious yawning remain unclear. Humans and dogs show a developmental increase in susceptibility to yawn contagion, with children showing an increase around the age of four, when also empathy-related behaviours and accurate identification of others’ emotions begin to clearly evince. Explicit tests of yawn contagion in non-human apes have only involved adult individuals and examined the existence of conspecific yawn contagion. Here we report the first study of heterospecific contagious yawning in primates, and the ontogeny of susceptibility thereto in chimpanzees,  Pan troglodytes spp.Pan paniscusTheropithecus geladaCanis lupus familarisMelopsittacus undulatusMacaca arctoides, elevated levels of concomitant self-directed scratching, suggested that the yawns derived from tension, rather than contagion Geochelone carbonariaContagious yawning  is well-established in humans feel and to imagine others’ emotional experiences. The ability to feel others’ emotional experiences (‘affective empathy’) derives from a phylogenetically old social contagion system, whereby one is viscerally affected by another’s emotional or arousal state , who seem primarily impaired in cognitive, but not affective, empathy cognitive empathy and theory-of-mind capacities  affiliate more with humans, who have previously imitated them per se. The ‘chameleon effect’ may nonetheless modulate CY, once the capacity is in place.Alternatively, CY may be unrelated to empathy An even lower level mechanism, a fixed action pattern familiar conspecifics , and emotional closeness to the yawning model. While attention levels are difficult to certify and quantify, we ensured that yawns were perceivable, by presenting them dependent on the chimpanzees’ attentional focus . We sought to increase ecological validity and the chance of evoking CY in younger subjects by using live rather than videoed models. For practical reasons we consequently used human models, and the study thus represents the first test of interspecies yawn contagion in primates. We hypothesised that, if CY is related to the development of empathy and emotional understanding in humans, a similar developmental effect might be found in chimpanzees. We therefore predicted that (1) juveniles would be more susceptible to CY than infants. As model identity and empathy may facilitate social behaviours, such as non-conscious facial mimicry Pan troglodytes verus), 12 infants  and 21 juveniles  that contained natural tropical forest foliage, edible plants and trees for climbing. The chimpanzees were housed in a number of smaller outdoor cages (25 m2) during the night. The remaining 21% (n = 7) were infants that had arrived at the sanctuary within 12 months and were housed either alone (n = 5) or in a dyad (n = 2). All the chimpanzees were experimentally naïve, and only animals, that did not exhibit overt signs of stress from the separation from conspecifics, participated in trials. One participant was excluded due to fussiness and one due to refusal to engage with the models.Participants were 33 orphaned chimpanzees  and two independent variables: model familiarity , and model behaviour . Each participant received seven 5-min phases presented in immediate succession . A trial sequence consisted of a baseline phase followed by three experimental phases, where the model repeatedly either yawned, gaped or nose-wiped. Each phase was followed by a five-minute post-stimulus observation interval, during which social interaction continued, without the inclusion of the key behaviours . The chimpanzees were tested, individually, between 8.30 h and 16.30 h in an outdoor cage, that otherwise served as their sleeping quarters. In all phases the model encouraged the participant to engage in calm interactions  through the bars of the enclosure.Yawning phase: The model repeatedly yawned while being within the participant’s ‘full’ or ‘peripheral’ field of vision . Models yawned as naturally as possible, with yawning defined as opening the mouth fully, drawing in air, lifting the shoulders, tilting the head and body backwards, closing the eyes, and producing a vocalisation during exhalation, for a total duration of 5–10 s.  mouth openings, i.e., opening the mouth widely and closing it, without audible inhalation and exhalation of air (approx. 4 s duration), while within the participant’s ‘full’ field of vision.Nose-wiping phase: The model repeatedly wiped her hand over her nose (approx. 3 s duration), while within the participants’ ‘full’ field of vision. = 0.13, P = 0.90), nose-wipes  = 1.34, P = 0.18), and gapes  = 0.57, P = 0.57) made by the two models did, however, not differ significantly, nor was there a relationship between the number of presented and elicited yawns  and nose-wipes . . All conditions (except for the baseline phase) were counterbalanced across participants, including trials with the familiar and unfamiliar models . The number of yawns . Values reported are mean ± SEM, and all tests were two-tailed and significance levels set at α = 0.05.Both individual phases  and the subsequent five-minute post-observation phases were used as means of comparisons . Trials from 21% of a random selection of participants were scored for interrater reliability, which was perfect with respect to number of chimpanzee yawns (100% agreement) and high with respect to nose-wipes (Cohen’s kappa = 0.94). We used independent t-tests to assess differences in the number of yawns, nose-wipes and gapes produced by models, and Spearman’s correlations to assess the relationship between numbers of presented and elicited yawns and nose-wipes. Data were analysed at both individual level  and group level. For the latter, the number of yawns and nose-wipes per minute were used, as the duration of the compared baseline, yawn, nose-wipe and gape conditions differed. Generalised linear mixed models (GLMM) were used to assess the effect of model behaviour, model familiarity and participant age on yawns and nose-wipes, and interaction effects. We controlled for participant identity (random effect) and used robust covariances and a Satterhwaite approximation . Friedman’s ANOVA and Wilcoxon signed ranks tests were used for post-hoc comparisons of number of yawns per minute across the baseline, yawn, gape and nose-wipe phases. Cochran’s Q and McNemar tests were used for binomial analyses. Moreover, Mann–Whitney Experimental procedures were non-invasive and complied with the ethical guidelines of the Pan African Sanctuary Alliance and Animal Behavior Society Guidelines for the Use of Animals in Research. The ethics of the experimental setup was evaluated by the Regional Ethics Committee at Uppsala District Court in Sweden, and the board of Tacugama Chimpanzee Sanctuary (in Sierra Leone) reviewed and approved the study.U = 66, z = 2.77, P = 0.006). Analysis of the number of yawns per minute revealed a significant effect of age , and an interaction between age and model behaviour , while model familiarity was non-significant . A difference in the number of juvenile yawns per minute across the baseline (0.00), yawn (0.06), nose-wipe (0.02) and gape phases  = 16.64, P = 0.001), reflected that juveniles were more likely to yawn in the yawn phase than all other phases . Both the familiar (Friedman test: X2(3) = 8.43, P = 0.038) and unfamiliar model (X2(3) = 13.02, P = 0.004) elicited significant differences in juvenile yawning across the conditions, though with slight differences in the patterns of results . Overall, across conditions, juvenile chimpanzees yawned at a higher frequency than infants . Moreover, juveniles produced twice as many yawns in the yawn phase  compared to the post-yawn phase , although the difference did not reach statistical significance .The yawn condition elicited 24 yawns from the juvenile chimpanzees and none from the infants, a statistically significant difference (Mann-Whitney U test: ults see . Juvenil(2) = 9.46, P = 0.009). Pairwise comparisons showed that more juvenile chimpanzees yawned in the yawn than gape phase (Q(1) = 2.95, P = 0.009), while the difference between the yawn and nose-wipe phase only approached significance (Q(1) = 2.22, P = 0.080). An equivalent number of juveniles yawned in the gape and nose-wipe conditions (Q(1) = 0.74, P = 1). The behaviour  of both the familiar and unfamiliar models elicited differences in the number of juveniles that yawned (Cochran’s Q: Familiar: Q(2) = 7.00, P = 0.03; Unfamiliar: Q(2) = 6.33, P = 0.042), though with slight differences. Pairwise comparison showed that the familiar model evoked yawns from more juveniles when she yawned than nose-wiped (Q(1) = 2.50, P = 0.037), while the unfamiliar model evoked more yawns when she yawned than gaped (Q(1) = 2.50, P = 0.037). In contrast, there was no difference in the number of juveniles that yawned in the yawn and gape phase when the behaviours were performed by a familiar model (Q(1) = 2.00, P = 0.137), and in the yawn and nose-wipe phases when the behaviours were performed by an unfamiliar model (Q(1) = 1.00, P = 0.952). Moreover, both the familiar and unfamiliar models evoked yawns in a similar number of juveniles in the gape and nose-wipe phases (familiar: Q(1) = 0.50, P = 1; Unfamiliar: Q(1) = 1.50, P = 0.401). An equal number of juveniles yawned to familiar and unfamiliar yawn stimuli (McNemar test: P = 1). Finally, yawn frequency did not differ between the sexes  and there were no order effects .Analysis of the number of chimpanzees the yawned (rather than the number of yawns per minute across the conditions) also revealed a significant difference in the number of juvenile chimpanzees that yawned across the yawn (48%), nose-wipe (19%) and gape phases , model familiarity , or age , although age approached significance . Moreover, juveniles produced more nose-wipes in the nose-wipe (mean = 9.43±1.16) than post-nose-wipe phase , while there was no similar effect for infants . With respect to the number of chimpanzees the nose-wiped across conditions (rather than the number of nose-wipes per minute), all but one chimpanzee (an infant), nose-wiped in response to the models’ yawns, nose-wipes and gapes, respectively. Moreover, equal numbers of chimpanzees nose-wiped when the familiar and unfamiliar models, respectively, yawned, nose-wiped and gaped (Cochran’s Q: familiar: Q(2) = 2.00, P = 0.368; unfamiliar: Q(2) = 2.00, P = 0.368).Analysis of the number of chimpanzee nose-wipes per minute revealed no effect of either model behaviour  is sufficient to explain the distribution of yawn contagion, ontogenetically and phylogenetically. The comparatively late emergence of CY relative to evidence of affective empathy in children below 4 years of age, suggests that CY may rely on the development of other and interacting capacities . While CY co-emerges temporally with cognitive empathy in children cognitive empathy in a species. A lower-level hypothesis suggests that susceptibility to yawn contagion relies on attentional states and biases Overall, the existence of CY in non-human species, and the development of CY in humans and other species, is consistent with the notion that the development of On a proximate level, the immunity of infant chimpanzees to CY may indicate a developmental immaturity of socio-cognitive skills and/or neural networks involved in processing social information In contrast to yawning, there was no contagion effect for nose-wiping. Juvenile chimpanzees were, however, more likely to wipe their nose during the 5-min interval, where they viewed a model nose-wipe, than in the subsequent 5 minutes . There was however, a (non-significant) trend for juvenile chimpanzees to nose-wipe at a higher frequency across all conditions than infants. Thus, while nose-wiping might be (low-level) contagious in humans (similar to itching unfamiliar (than familiar) model may suggest that the ‘chameleon effect’ model familiarity . Further research is thus required to ascertain the mechanisms underlying the variable results in studies of CY across ages and species.Contrary to prediction, but consistent with findings for human children Table S1Participant details .(DOCX)Click here for additional data file.Table S2Order of presentation of the conditions (model familiarity and model behaviour).(DOCX)Click here for additional data file.Video S1Baseline, yawning, nose-wiping and gaping conditions with the familiar and unfamiliar models .(M4V)Click here for additional data file.


Epidemiological studies tend to correlate a PUFA-rich diet with a reduced incidence of cancer, including breast cancer. However, the molecular and cellular mechanisms supporting the effect of PUFAs in breast cancer cells remain relatively unknown. Here, we review some recent progress in understanding the impact that PUFA may have on breast cancer cell proliferation, apoptosis, migration, and invasion. While most of the results obtained with docosahexaenoic acid and/or eicosapentaenoic acid show a decrease of tumor cell proliferation and/or aggressivity, there is some evidence that other lipids, which accumulate in breast cancer tissues, such as arachidonic acid may have opposite effects. Finally, lipids and especially PUFAs appear as potential adjuvants to conventional cancer therapy.For several years, lipids and especially  Epidemiological studies have shown that the rate of breast cancer is 4 to 5 times higher in Western countries that in Japan , 2 suggemergence –9.n − 3 PUFAs on breast cancer incidence yielded mixed results, and most of them did not show a significant association between n − 3 PUFAs consumption and breast cancer risk [However cohort studies that examined the effect of cer risk , 11. Nevcer risk . Then, in − 3 PUFAs, namely, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) have demonstrable anticancer properties both in vitro and in vivo. However, the mechanisms behind the benefits are not clear [Several authors have shown that ot clear –16. ThisSeveral studies showed that DHA and EPA together or alone inhibit the growth of breast cancer cell lines –19. SomeActivation of the p44/42 mitogen-activated protein kinase (MAPK) pathway plays a major role in regulating cell growth and survival in breast cancer cells  and is pn − 3 PUFAs in membranes decreased arachidonic acid (AA) content and n − 6/n − 3 PUFA ratio in the membranes, without modifying the unsaturation index [n − 3 PUFAs [n − 3 PUFA-treated cancer cells associated with growth arrest and apoptosis [n − 3 PUFA-induced apoptosis suggesting the involvement of lipid peroxidation-derived ROS [Incorporation of on index . Consequ 3 PUFAs , 30. Whe 3 PUFAs , severalpoptosis –38. Somepoptosis . Glutathpoptosis , 35. Thepoptosis  whereas poptosis , 37. Theived ROS –38. Then PUFAs appeared as proliferation inhibitors and apoptosis inducers in breast cancer cell lines and animal models, at least in part through MAPK and ROS pathways , 39.n − 3 PUFAs may exert their growth inhibitory effects on cancer cells by altering the plasma membrane composition and associated signaling events [The g events . One emeg events –45. Modeg events , 46, 47.g events . n − 3 PUFAs on lipid rafts and their signaling pathways has been studied in several cell types with cancerous origin or not [n − 3 PUFAs exposure resulted in a decreased level of the chemokine receptor CXCR4, which requires intact lipid rafts for signaling [The effect of n or not . Particun or not , 50. In ignaling . Schley ignaling . These aignaling . Increasignaling –55, and ignaling , 56. Furignaling . These dignaling , 57.Corsetto et al. have recently examined the PUFA incorporation in breast cancer lipid rafts and showed that PUFA are incorporated preferentially in phosphatidylinositol, phosphatidylserine, and phosphatidylcholine that may be relevant to the formation of biologically active metabolites such as prostaglandins, prostacyclins, leukotrienes, resolvines, and protectines . These an − 3 PUFA-mediated alteration of lipid rafts was linked to oxidative stress modulation. The treatment of rat hepatocytes with EPA increases ethanol-induced oxidative stress via lipid raft aggregation and subsequent phospholipase Cγ translocation into these microdomains [n − 3 PUFA-mediated remodeling of lipid rafts in breast cancer cells with respect to oxidative stress? Recently odomains . EPA incodomains . In a diodomains . Then whn − 3 PUFA [β-cyclodextrin reduced uPAR and matrix metalloproteinase-9 (MMP-9) colocalization in lipid rafts and inhibited breast carcinoma cell migration and invasion. The decreased expression of uPAR and MMP-9 was reversed by cholesterol supplementation [+ channels [+ channels  in a dose-dependent manner, and tetrodoxin, a compound that specifically blocks this type of channels, reduces MDA-MB-231 cell migration at the same level that observed in the presence of DHA [+ channel mRNA and functional protein expression [+ channels localization in lipid rafts, such as shown in cardiac cells [Because metastasis is the leading cause of death from breast cancer, reducing the invasive potential of breast cancer cells is almost as important as destroying in the primary tumor. Recently, we showed that DHA reduces the invasive potential of the MDA-MB-231 breast cancer cell line . Interes− 3 PUFA , are cri− 3 PUFA . In thisentation . In addichannels , 64. Indac cells , may be ac cells .n − 3 PUFAs results in decreased surface levels of CXCR4 in a time- and dose-dependent manner [n − 3 PUFA treatment [One of the hallmarks for breast cancer metastasis is the over expression of chemokine receptors, which leads to migration of the cancerous cells to surrounding tissues –68. The t manner . Migratireatment . These dreatment . −) breast cancer cells [+ MCF-7 cell line leads to the investigation of neutral lipid composition and metabolism in these cells [+ cells [−. Interestingly, LDL was shown to stimulate proliferation of ER-cells in a manner dependent from ACAT1 activity [Some recent studies focused on the negative effect lipids may have on breast cancer evolution and/or emergence. Both LDL and unsaturated fatty acids have been demonstrated to increase proliferation of estrogen receptor alpha negative  in correlation with PPARα expression and activity [β-activated kinase-1 [α, Src, Erk1/2, and lipoxygenase activities [Examining fatty acid composition of breast cancer tissues Chang et al. observed higher levels of AA, stearic acid, and DHA in tumoral tissue by comparison with healthy tissue of the same patient . They al tissues . Further tissues –84. Besiactivity . Invasivkinase-1  and ubiqkinase-1 . Treatmetivities . The mamtivities . This mTtivities . In additivities .κB activation, matrix metalloproteinase-2 and -9 secretion, and finally cell migration and invasion [γ and E-cadherin/β-catenin pathway and/or estrogen receptor and protein phosphatase-2A depending of the cell type [n − 3 PUFAs.Like AA, linoleic acid promotes the proliferation and the migration of cancer cells. Linoleic acid induces expression of plasminogen activator inhibitor-1, proliferation, migration, and invasion of breast cancer cells , 91. Lininvasion . By continvasion . CLA hasinvasion –97. Breaell type , 99. Cavell type , leadingn − 6 PUFAs, such as linoleic acid and AA have a tumor promoting effect [n − 6 PUFAs is through the production of proinflammatory eicosanoids such as prostaglandin E2, which promotes angiogenesis and hinders apoptosis. Alternatively, n − 3 PUFAs, such as EPA and DHA are the precursor molecules to eicosanoids that are less inflammatory when compared to AA-derived prostanoids [n − 6 PUFAs may be contributing to the high risk of breast cancer in the US, and that n − 3 PUFAs may have a protective effect [n − 6 PUFA intake, n − 3 PUFA intake, and breast cancer risk. Women with lower intake of n − 3 PUFA and higher intake of n − 6 PUFA had an increased risk for breast cancer compared to women with higher intake of n − 3 PUFAs and lower intake of n − 6 PUFAs [n − 6 fatty acid consumption among subjects who consumed low levels of marine n − 3 fatty acids [n − 6 PUFA to n − 3 PUFAs may be more important for breast cancer risk than individual dietary amounts of these fatty acids.Animal models have demonstrated that long chain PUFAs have differential effects on mammary tumorigenesis based on double bond position and meta-analyses of mouse mammary tumor models have suggested that g effect –103. A pstanoids , 104. Ine effect . A recen 6 PUFAs . Accordity acids . To concn − 3 PUFA exert little or no cytotoxic effects on normal breast cells [n − 3 PUFA may now be considered as powerful nontoxic adjuvant of canonical anticancer treatments [in vitro observed effects will be translated in vivo. Also, it will be important to determine the ratio of n − 3 versus n − 6 PUFAs needed to be consumed for the beneficiary effect to be achieved.High fat diets, obesity, and metabolic syndrome leading to high circulating level of lipoproteins are harmful for more than one aspect, but they may also be aggravating for breast cancer by stimulating tumor proliferation and metastasis. On the contrary PUFAs are generally regarded as safe compounds that are well tolerated and produce few side effects. Their effects on nontumorigenic cells have not been fully elucidated, but some studies suggest that when provided at concentrations that inhibit tumor cell growth, st cells , 18, 108st cells , 110. DHst cells . Then n eatments , especia


This paper explores self-perceived mate value (SPMV), and its association with self-esteem, in eight cultures. 1066 participants, from 8 cultural groups in 7 countries, rated themselves on 24 SPMVs and completed a measure of self-esteem. Consistent with evolutionary theory, women were more likely to emphasise their caring and passionate romantic nature. In line with previous cross-cultural research, characteristics indicating passion and romance and social attractiveness were stressed more by respondents from individualistic cultures, and those higher on self-expression  values; characteristics indicative of maturity and confidence were more likely to be mentioned by those from Traditional, rather than Secular, cultures. Contrary to gender role theory, societal equality had only limited interactions with sex and SPMV, with honesty of greater significance for male self-esteem in societies with unequal gender roles. These results point to the importance of cultural and environmental factors in influencing self-perceived mate qualities, and are discussed in relation to broader debates about the impact of gender role equality on sex differences in personality and mating strategies. A great deal of previous research has examined the partners we seek for a romantic relationship Work on perceived mate values has been influenced by two theoretical traditions. From an evolutionary perspective, each species has a genetically organized set of strategies and tactics for survival, growth and reproduction A second theoretical perspective, social structural theory From both theoretical perspectives, perceived qualities have significant implications for self-esteem. According to sociometer theory At present, the great majority of the research on SPMV, and SPMV and self-esteem, has been conducted in the U.S. However, as noted above, both social structural theorists and evolutionary theorists recognize that social and ecological conditions influence the choice and evaluation of partners We complement this with a second set of cultural dimensions - Survival values vs. Self-Expression values and Traditional vs. Secular values - from the World Values Survey Previous work on SPMV has focused on research conducted in Western cultures, and has failed to explore cross-cultural variations in these values. In addition, most research has focused on the exchange of female beauty and male resources, but SPMV is a multi-faced concept that includes more than women's beauty or male resources M age 24.0, SD = 7.7) from eight different cultural groups. Two samples were collected in Britain, from British White and British Asian participants in the same London University. Further data were collected in major urban Universities in Ghana, Portugal, Poland, China, Hungary and Spain. All participants were recruited on the campuses during class time or following classes. Participants were 1066 students . Self-esteem was assessed using the 10-item Rosenberg Self-Esteem Scale, the most widely used and validated self-report measure of global self-esteem Pilot work at a London University generated 24 commonly occurring qualities that thought they possessed that a romantic partner might find attractive. The 24 item qualities obtained were then used to create an ‘SPMV index’. Participants were asked the extent to which they possessed each of the 24 characteristics ; socially attractive ; passionate romantic  adventurer  and mature confident .Principal components analysis (with varimax rotation) reduced the 24 SPMV characteristics to five factors, which explained together 45% of the variance: socially attractive  = 4.22, p<.04; with men higher on this dimension) and passionate romantic  = 12.99, p<.001, with women higher on this dimension). There were also unique culture grouping effects for the SPMV dimensions of caring  = 54.97, p<.001), socially attractive  = 102.05, p<.001), mature confident  = 26.46, p<.001), and passionate romantic  = 63.77, p<.001). Participants from individualistic cultures were higher on passionate romantic  vs. 6.84 (collectivist)), caring  vs. 22.89 (collectivist culture)), socially attractive  vs 19.22 (collectivist culture)) and mature confident  vs. 19.84 (collectivist)). In addition, there was a sex x cultural grouping interaction on the mature confident dimension, with this dimension score highest amongst males from individualist cultures (M = 21.47), and lowest for males from collectivist cultures (M = 19.38)We first considered the impact of culture and sex on the five SPMV factors. To meaningfully examine these cultural effects we employed the three cultural dimensions described above . Because these cultural dimensions overlap conceptually, we conducted three separate MANOVAs, with the cultural dimensions and sex as the independent variables and the five SPMV factors as criterion variables. Below we report the significant effects. In our first analysis, with Individualism/Collectivism as the cultural grouping, there were sex effects for the SPMV dimensions of caring dimension , F = 5.26, p<.02) and for passionate romantic scores  = 19.95, p<.001), with females higher on caring (M = 24.41 (women) vs. 23.88 (men))) and passionate romantic (M 7.73 (women), 7.23 (men)). There were unique culture effects for mature confident  = 32.37, p<.001) and passionate romantic  = 10.21 p<.001). In both cases traditional cultures scored higher on these dimensions  vs. 20.03 (secular culture); for passionate romantic  vs. 7.30 (secular)). There were also cultural grouping x sex effects for caring  = 14.71 p<.001), socially attractive  = 5.36 p<.02), mature confident  = 9.40 p<.002) and passionate romantic  = 20.19, p<.001). Highest on caring were women from secular cultures (M = 24.75), lowest on this dimension were men from secular cultures (M = 23.32). Highest on socially attractive were men from traditional culture (M 21.35), lowest were men from secular cultures (M = 20.68). Men from traditional cultures scored highest on mature confident (M = 21.70), men from secular cultures lowest on this dimension (M = 19.63). Passionate romantic was highest amongst females from secular cultures (M = 7.80), lowest for males from these cultures (M = 6.80).We then turned to the Tradition-Secular dimension and Security-Self Expression cultures. Here there were unique sex effects for the caring  = 5.25 p<.02, M = 24.55 (women) vs. 24.02 (men)), and passionate romantic  = 17.93, p<.001, M = 7.77 (women) vs. 7.30 (men)). There were unique culture effects on caring  = 68.92, p<.001), socially attractive  = 10.17, p<.001), mature confident  = 101. 91, p<.001) and passionate romantic  = 48.55, p<.001). Self expressive cultures scored higher on caring (Ms = 25.24 (self expressive) vs. 23.33 ), socially attractive (respective Ms = 21.40 vs. 20.63), mature confident (respective Ms = 22.03 vs 19.69) and passionate romantic (respective Ms = 7.92 vs. 7.15). There were no sex x culture interactions.For our final dimension, security-self expression, there were unique sex effects, with women higher on r = .38). We conducted three multiple regressions, one for each cultural dimension , with self-esteem as the criterion variable. Predictors were the five centered SPMV factors, cultural grouping  and the interaction between SPMV dimension and cultural group. Below we report the significant effects. Socially attractive, mature confident, and passion romantic all significantly correlated with self-esteem . There was no unique effect for Individualism/Collectivism on self esteem but an interaction between SPMV and cultural group, with mature confident more highly correlated with self-esteem for respondents from individualistic, versus collectivist, cultures . Those in Secular cultures enjoyed a greater degree of self-esteem  vs. 3.60  cultures), with caring more strongly correlated with self-esteem in traditional cultures . Comparing survival and self-expressive cultures, there was a unique effect for cultural grouping on self-esteem , with those in self-expressive cultures enjoying higher self-esteem (Ms = 3.95 (expressive) vs. 3.52 ). Those who scored more highly on caring in survival–oriented cultures also scored higher on self-esteem .Turning to the relationship between the SPMV factors and self-esteem, all eight cultural samples demonstrated a significant moderate correlation between total SPMV scores and self-esteem (ranging from .23 (White British respondents) to .49 (Polish sample), average sample p<. 01) and there was a unique effect for gender role differentiation, with the more equal cultures higher on self-esteem than the most unequal cultures . There was a two way interaction for SPMV x sex for the caring factor , with men more strongly correlating caring with their self-esteem. However, a three-way interaction between caring x sex x sex role grouping also demonstrated greater sex differences in the relationships between esteem and this attribute in the more sex role unequal cultures . Hence while the correlation between self-esteem and caring was small for both men and women in the more sex-role equal societies (r (99) = .03 for men; r (164) = .07 for women), men were more likely to correlate esteem and caring in the more unequal sex-role cultures (r (99) = .49 and r (102) = .16 for men and women, respectively). A further analysis of the caring factor demonstrated that it is on the single SPMV attribute ‘honesty’ that the sex/culture groups most clearly differ. In the unequal sex-role societies it was men who most clearly related this attribute to their self-esteem (r (103) = .34 for men, r (104) = .05 for women, in the most unequal societies). In contrast, honesty was not closely correlated with self-esteem for either sex in the more equal gender-role societies (r (85) = −.04 for men, r (182) = −.10 for women). Finally, there was a three-way interaction between sex, the adventurer category of SPMV, and the contrast between gender egalitarian and ‘moderately’ gender equal cultures. In moderately egalitarian cultures, women's score on adventurer was more strongly correlated with their self-esteem (r (231) = .17 for men, r (354) = .33 for women).Finally, we predicted a sex x cultural sex role egalitarianism interaction in the relationship between SPMV and self esteem. We created two new dummy variables, the first contrasting the more sex role egalitarian cultural groups  with the most unequal cultures ; the second contrasting the gender equal and the other ‘medium’ scores on GDI . We ran a multiple regression with self-esteem as the criterion variable and with the five SPMV factors as predictors, alongside sex, the dummy codes for sex role grouping, sex x sex role grouping, each SPMV x sex, each SPMV by sex role group, and the three way interactions for each SPMV by sex by sex role group. Men scored higher on self-esteem overall  across eight cultural groups in seven countries, and the relationship between perceived mate value and self-esteem by gender and culture. When asked to rate key attributes for attracting a mate, women were more likely to emphasise their caring and passionate romantic side in their self-ratings of attractiveness factors. These finding are largely consistent with evolutionary theories, which emphasise women's greater caring role and emotional investment in relationships caring dimension demonstrated that it was a self-perception of faithfulness which was more important for male self esteem (respective rs of .24 (men), .09 (women)), while supportiveness was more significant for female esteem ). There was no consistent significant sex difference in self-ratings of the broader factor of socially attractive, which consisted of both appearance but also a range of personality qualities. This failure to find sex differences in self-perceived attractiveness has been reported elsewhere First, let us consider sex differences in the attributes emphasized by our participants. As predicted, women were keen to emphasise their passionate romantic and socially attractive  were less stressed by respondents from collectivist cultures. Mature confident was also more likely to be mentioned by those from Traditional, rather than Secular, cultures, as hypothesized. The BSD evolutionary model socially attractive and passionate romantic was more frequently mentioned as attractive characteristic possessed by our respondents. While our participants were more likely to describe themselves as caring in individualist and self-expressive societies, caring was positively correlated with self-esteem in the more traditional and security-orientated cultures. Thus while being humorous, cultured or sociable may be particularly important in richer societies, where survival obligations are less prominent, being able to care for others may be important for esteem where economic survival is threatened.As discussed above, both social role and evolutionary perspectives recognize that there can be important cultural variations in the qualities valued in a partner. As anticipated, participants in individualist societies were more likely to highly rate qualities associated with emotional investment in a relationship caring scoring higher on self-esteem in less egalitarian cultures. This finding resulted primarily from differences in the single attribute of ‘honesty’, with honesty of greater significance to men in sex unequal societies than elsewhere. Given that the sexual infidelity of men may of particular issue in cultures where there is greater gender inequality (such as Ghana), the issue of honesty in relationships may be more predominant in such societies caring dimension (reported above), this finding suggests that ‘caring’ as an attribute has a number of different dimensions, with both culture and gender likely to influence their relative importance. Further, this lack of wider systematic differences between egalitarian and more sex-role unequal societies in our data is perhaps less surprising given apparently conflicting recent evidence on societal equality and sex differences in sociosexual orientation and personality traits From a social role perspective, we would expect role equalities in the division of labour to influence the qualities emphasized by individuals as attractive to mates, and the relationship between these qualities and self-esteem. Here we received only mixed support for the impact of societal gender-role differentiation on the relationship between self-perceived mate attributes and esteem. Contrary to our expectations, there was only one interaction effect between perceived attributes, societal equality and sex, with men who scored more highly on Xiao) is an important factor in mate choice The present study had several limitations. Our respondents were largely young students from urban areas. Across the lifespan, mate value generally declines for women and increases for men Lastly, what are the broader implications of SPMV across cultures? The findings of our study make new contributions to our understanding of our self-perceived mate value, those qualities that make us feel positive about ourselves, and how this varies across sex and cultural context. However, just as particular beliefs might seek to aid or abet such relationships in particular settings


Generally, chemical tissue clearing is performed by a solution consisting of two parts benzyl benzoate and one part benzyl alcohol. However, prolonged exposure to this mixture markedly reduces the fluorescence of GFP expressing specimens, so that one has to compromise between clearing quality and fluorescence preservation. This can be a severe drawback when working with specimens exhibiting low GFP expression rates. Thus, we screened for a substitute and found that dibenzyl ether  can be applied as a more GFP-friendly clearing medium. Clearing with dibenzyl ether provides improved tissue transparency and strikingly improved fluorescence intensity in GFP expressing mouse brains and other samples as mouse spinal cords, or embryos. Chemical clearing, staining, and embedding of biological samples mostly requires careful foregoing tissue dehydration. The commonly applied tissue dehydration medium is ethanol, which also can markedly impair GFP fluorescence. Screening for a substitute also for ethanol we found that tetrahydrofuran (CAS 109-99-9) is a more GFP-friendly dehydration medium than ethanol, providing better tissue transparency obtained by successive clearing. Combined, tetrahydrofuran and dibenzyl ether allow dehydration and chemical clearing of even delicate samples for UM, confocal microscopy, and other microscopy techniques. In the recent years, GFP has become a common reporter gene in molecular biology, medicine, and cell biology Light microscopy techniques, as confocal microscopy THF is a colorless, flammable, cyclic ether with an ethyl ether like odor Figure 1To assess the compatibility of GFP with THF dehydration, the left hemispheres of two GFP-expressing paraformaldehyde fixed mouse brains  is slightly higher than the refractive index of BABB (n = 1.559).Phenylmethoxymethylbenzene   is a colorless cyclic ether  mice) with THF. The two left hemispheres were cleared with peroxide free DBE and the corresponding right hemispheres were cleared with peroxide free BABB as a control. As illustrated by We further prepared isolated mouse hippocampi n = 1.518), benzyl alcohol (n = 1.540), benzyl benzoate (n = 1.569), trans-anethole (n = 1.561), isoeugenol (n = 1.578), isosafrol (n = 1.573), 1,5-dibromopentane (n = 1.513), and bromobenzene (n = 1.560) . As in the experiments before, the left hemispheres of mice brains were treated with the respective test substance. The corresponding right hemispheres were treated with ethanol/BABB as a control.In order to find a more GFP-friendly tissue dehydration medium, we further checked methanol, acetone, 2-butoxyethanol, dimethylformamide, dimethylsulfoxide (DMSO), and dioxane. For GFP-friendly chemical clearing, we further tested methylsalicylate  until all blood was removed, followed by 100 ml 4% Paraformaldehyde in 0.1 M PBS for perfusion-fixation. We removed the brains from the skull, and placed them in the fixative for overnight at 4°C, rinsed them 3 times in PBS (30 min each). Animal care and euthanasia was done in accordance with the ethic guidelines of the German Max-Planck society and with the Austrian animal protection law. According to §2 of the Austrian animal experiments act, special approval by an ethics committee or an approval number was not required, since all experiments were performed on isolated organs and not on living animals.Six mouse brains were obtained from 3–5 weeks old thy-1 EGFP-M (c57/bl6) mice The fixed brains were split along the median line into their left and right hemispheres. The hemispheres were differently incubated in ascending concentration series of a) ethanol, or b) peroxide free THF . The applied concentration steps were 50 Vol%, 70 Vol%, 80 Vol%, 96 Vol%, 3×100 Vol% for ethanol as well as for THF (12 hours per step).The dehydrated left or right hemispheres were incubated at room temperature in separate glass vials, each containing ∼10 ml peroxide free BABB or peroxide free DBE, respectively. DBE was obtained from Sigma-Aldrich, Austria (Order-No. 108014). The specimens were left in the clearing solution until they became transparent (about 1–2 days). During incubation, the clearing medium was exchanged three times. For better diffusion, the specimen vials were slightly shaken on a vibrating table placed under an extraction hood.Mouse brains were prepared as described above. The hippocampi were dissected and stored in 4% PFA for post fixation overnight. Afterwards they were rinsed three times in PBS (15 min each) and immediately dehydrated.The dissected and fixated hippocampi were incubated in ascending concentration series of peroxide free THF. The applied concentration steps were 50 Vol%, 80 Vol%, 96 Vol%, 100 Vol% THF .The dehydrated dissected mouse hippocampi were incubated in BABB or DBE until they became transparent (about 3–6 hours).Peroxide removal in THF was done by column absorption chromatography (Due to their higher viscosity and boiling points, removal of peroxides in BABB and DBE can be done using the setup described in


A receptors and spillover onto perisynaptic GABAA receptors, resulting in increased tonic inhibition. Recent studies show that the brain can engage in a limited process of neural repair after stroke. Changes in cortical sensory and motor maps and alterations in axonal structure are dependent on patterned neuronal activity. It has been assumed that changes in neuronal excitability underlie processes of neural repair and remapping of cortical sensory and motor representations. Indeed, recent evidence suggests that local inhibitory and excitatory currents are altered after stroke and modulation of these networks to enhance excitability during the repair phase can facilitate functional recovery after stroke. More specifically, dampening tonic GABA inhibition can afford an early and robust improvement in functional recovery after stroke.An attempt to find pharmacological therapies to treat stroke patients and minimize the extent of cell death has seen the failure of dozens of clinical trials. As a result, stroke/cerebral ischemia is the leading cause of lasting adult disability. Stroke-induced cell death occurs due to an excess release of glutamate. As a consequence to this, a compensatory increased release of GABA occurs that results in the subsequent internalization of synaptic GABA A, GABAB, and GABAC receptors [ARs are ligand-gated chloride channels [AR subunits exist: α1–6, β1–3, γ1–3, δ, ε, φ, π, and ρ1–3, with slice variants also contributing to variations in receptor functions [α, 2β, and γ, with the γ-subunit being able to be substituted for either an ε- or a δ-subunit [GABA is the major inhibitory neurotransmitter within the mammalian brain. Twenty to 50% of all synapses within the CNS use GABA as a neurotransmitter, mediating both fast and slow inhibitory synaptic transmission . GABA iseceptors , and thechannels , 3 formeunctions –9. The m-subunit –9. AR's respond to ambient levels of GABA present in the extracellular space to regulate baseline pyramidal neuron excitability and show reduced desensitization remaining active for long periods of time [AR's in the hippocampus and cortex contain either α5 or δ-subunits [α5 or δ-subunits enhances pyramidal neuron firing to afferent inputs [AR activity [AR's, leading to persistent or tonic inhibition [AR's exhibit distinct pharmacological and biophysical properties that differentially influence brain physiology and behavior [Depolarization of inhibitory interneurons produces a phasic release of GABA and inhibition of postsynaptic neurons. Extrasynaptic GABA of time . Tonic Gsubunits , 10. Redt inputs , enhancet inputs , and fact inputs –17. GABAactivity  with thehibition , 20. Synbehavior .AR's are composed of α1–3, β1–3, and γ1–3, subunits, and the site of action for a variety of clinically important drugs, such as benzodiazepines, neurosteroids, and anesthetics. Where as extrasynaptic GABAAR's are composed of subunit combinations containing α4–6, β1–3, and γ2- or δ-subunits. Of these receptors, the δ-containing GABAAR's coassembled as α4βδ—located in the cortex, hippocampus and thalamus—or α6βδ—located in the cerebellum—that are emerging as unique and fundamental players in GABAergic neurotransmission [δ-containing GABAAR's having a functional role in the cortex, the α5-containing GABAAR's coassembled primarily as α5βγ2 have also been implicated in poststroke repair [α5-subunit is low in the cortex compared to the δ-subunit, greater functional improvements in motor recovery are seen following modulation of the α5-subunit [Synaptic GABAsmission . In addie repair . Even th-subunit . The pha-subunit  and has -subunit . Conflic-subunit , 24. DetStroke is the leading cause of death and long-term disability in adults worldwide. Stroke-induced sensory and motor loss of limb function, in particular, prevents patients from returning to work and accounts for the statistic that almost one-third of stroke survivors become institutionalized after having a stroke –28. Receγ-aminobutyric acid GABA activity within the motor cortex could facilitate structural changes [Functional recovery within the peri-infarct cortex involves changes in neuronal excitability. Clinical studies using direct current stimulation of the peri-infarct cortex, with protocols that boost local neuronal excitability, have been shown to improve use of the affected limb in stroke patients , 33. Fur changes  and prom changes . Alterat changes  such as  changes . Further changes . These dα-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor currents can promote poststroke function gain [The processes of neurorehabilitation involve physical, occupational, and cognitive therapies , 28. Furion gain , 39. Thiion gain , 41. In ion gain .α5GABAAR negative allosteric modulators are part of a broad class of drugs that boost learning and memory function by influencing key elements in neuronal memory storage, such as LTP [α5GABAAR negative allosteric modulators, and indeed any mechanism that dampens tonic GABA signaling, could significantly improve poststroke recovery [Modulation of learning and memory pathways have previously been shown to promote functional recovery and poststroke axonal sprouting following administration of pharmacological agents such as amphetamines and phosphodiesterase type-4 inhibitors that boost cAMP/CREB signaling and learning and memory function , 44. Theh as LTP , 16. α5Grecovery . This suMost strategies that promote functional recovery after stroke, such as axonal sprouting, neurogenesis, or angiogenesis, focus or rely on inducing structural changes in the brain as a means to promote functional recovery after stroke –53. In oAs with stroke recovery, the processes of learning and memory can be enhanced by manipulations that increase neuronal excitability. However, unlike the stroke recovery field, basic science studies in learning and memory have defined specific cellular pathways that lead to enhanced neuronal excitability and improved function.α5 and δ-subunit containing GABAAR's. Recent evidence using α5GABAAR “knock-out”, and point-mutated mice have clearly shown that the α5-subunit plays a key role in cognitive processing [in vitro and in vivo work has shown that α5GABAAR negative allosteric modulators can enhance cognition within the Morris water maze, enhance hippocampal LTP and do not have any proconvulsant effects [AR's, we have shown marked improvements in functional recovery when starting treatments from 3 days after the stroke [AR's in processes involving synaptic plasticity and learning and memory and more recently poststroke recovery.Recent work has shown that enhanced neuronal excitability occurs following the dampening of the baseline level of inhibition in neurons. This baseline inhibition is in part set by a tonic, always present, degree of inhibitory signaling from the major inhibitory neurotransmitter, GABA. Unlike the phasic nature of synaptically released GABA, the action of GABA via extrasynaptic receptors is to tonically suppress neuronal excitability and to help regulate neuronal action potential firing. These extrasynaptic GABA receptors consist of ocessing , 17. In  effects , 16. Usie stroke . These dNeuronal inhibition and network function is disturbed in peri-infarct tissue during periods of cortical plasticity, re-mapping, and recovery. The increase in tonic inhibition in cortical pyramidal neurons reported by Clarkson and colleagues  occurs aAR neurotransmission provides an alternative means for neuroprotection. Indeed, over recent years, changes in GABA function following cerebral ischemia and possible protective benefits of GABAergic drugs have been extensively assessed [α1 subunit GABAAR modulator, can result in transiently improves in aphasia in chronic stroke survivors [A large body of work has been devoted to developing and exploring neuroprotectants that act to block glutamate-mediated neurotransmission in animal models of cerebral ischemia , 58. Incassessed –65. Evenassessed –62, 65, assessed –65. Howeassessed , 67. Theurvivors .ARs to high concentrations of GABA agonists in vitro has routinely been shown to become desensitized and/or downregulated [AR is also downregulated in the gerbil hippocampus following transient cerebral ischemia [AR's resulting in an increase in tonic inhibition. Indeed, recent evidence showing an increase in tonic inhibition after stroke supports this notion [During situations of cerebral ischemia, it has been shown that the extracellular concentrations of GABA increase  to the micromolar range , 69 and egulated –72. Simiischemia . In thisischemia . This ins notion . This inChanges in neuronal excitability, loss of GABAergic inhibition, enhanced glutamatergic transmission, and synaptic plasticity all contribute to neuronal reorganization after stroke. Studies that promote an increase in local brain excitability result in improved function , 39, 45 AR's to tonically suppress neuronal excitability and regulate neuronal action potential firing. Therefore, in order to facilitate functional recovery, an increase in brain excitability is required to overcome this hypofunctionalism [AR's, implicating α5 or δ-containing GABAAR's as novel targets for developing agents to help stroke sufferers.Tonic or continuous signaling from GABA sets baseline inhibition. GABA acts via extrasynaptic GABAionalism . RecentlARs have been shown to mediate excitation as well as play an important role in neural migration and synaptogenesis [A receptor desensitization and/or downregulation [α1, α2, α3, α5, and γ2 subunits following photothrombotic stroke and freeze-lesion-induced cortical injury [GABA has been shown to mediate both fast and slow inhibitory synaptic transmission . During ogenesis , 74. Durogenesis , 69, resgulation , 71. Thil injury –77.α5GABAARs and results in an increase/substitution of other GABAAR's with a subsequent increase in rather than suppression of tonic inhibitory currents [α4δ-mediated tonic currents has been seen in the α5 knockout mice within region CA1 of the hippocampus [α5 or δ-mediated increase in tonic GABA currents, results in early and marked improvements in functional recovery [Recent work has shown that epileptogenesis results in the suppression of functionally active currents . A similpocampus . Extracepocampus . Consistpocampus . This porecovery .α1 and β2 GABAAR subunits are densely localized within the cortex and coassembly with the γ2-subunit accounts for about 40% of all GABAARs within the cortex [ARs containing α1β2γ2 has been shown to be enriched at synaptic sites throughout the cortex [δ subunit can coassemble with α1 subunits to form functional recombinant receptors[δ subunits can associate with α1 subunits [ARα1 subunits have also been found extrasynaptically [δ-containing GABAARs8181. These data could suggest an alternative method for why zolpidem was having an effect in chronic stroke patients to alleviate the burden of aphasia. However, further studies are needed, as one previous study would suggest that the γ2-subunit is required in order for zolpidem to have an effect [Understanding the profile for which cortical plasticity occurs, altered after a stroke, is critical for fully determining when to start treatments and with what therapeutic compound to use. Based on our findings, we have clearly shown that dampening of tonic GABA currently from 3 days results in robust functional improvements of motor recovery . These ie cortex . Assemble cortex  and invoreceptors, 83. Fursubunits , and GABptically , 86 consn effect .AR negative allosteric modulator, L655-708, exacerbated the lesion [Disinhibition of cortical connections within the peri-infarct or regions associated with the peri-infarct cortex have been argued as either occurring as a direct consequence of the stroke or as a potential compensatory mechanism related to the recovery . This are lesion . Furthere lesion , providee lesion . Given te lesion . This ise lesion . The pere lesion , which we lesion .AR antagonists [ex vivo and promotes recovery of motor function in vivo. These effects occur through blockade of α5 or δ-containing GABAAR's. This data indicates a novel role for tonic GABAAR function in promoting poststroke recovery most likely via cortical disinhibition [Therapies that promote functional recovery after stroke are limited to physical rehabilitation measures. While specific measures, such as constraint-induced therapies, promote recovery of motor function, no pharmacological therapies are available that aid in recovery. Functional recovery after stroke follows psychological learning rules  that indagonists . Recent hibition , 92, 93


The skin provides an effective physical and biological barrier against environmental and pathogenic insults whilst ensuring tolerance against commensal microbes. This protection is afforded by the unique anatomy and cellular composition of the skin, particularly the vast network of skin-associated immune cells. These include the long-appreciated tissue-resident macrophages, dendritic cells, and mast cells, as well as the more recently described dermal γδ T cells and innate lymphoid cells. Collectively, these cells orchestrate the defense against a wide range of pathogens and environmental challenges, but also perform a number of homeostatic functions. Here, we review recent developments in our understanding of the various roles that leukocyte subsets play in cutaneous immunobiology, and introduce the newer members of the skin immune system. Implications for human disease are discussed. The skin is a unique organ that serves as an interface between the host and the environment, providing a mechanical and biological barrier against chemical, physical and pathogenic insults. Anatomically, the skin comprises two distinct compartments: the epidermis, an avascular layer mainly composed of keratinocytes, and the dermis, a fibroblast-rich network of collagen and elastin fibers that provides the skin with strength and elasticity. The dermis also contains capillary and lymphatic vessels, which serve as the entry and exit portals for immune cells. Additional skin appendages such as hair follicles, sebaceous glands and sweat glands, as well as nerve endings are also found in the dermis . The skiMacrophages represent a key sentinel population for invading pathogens and tissue damage. These cells also perform developmental and homeostatic functions . Much ofMacrophages comprise a remarkably diverse and heterogeneous population that is found in most tissues, including the skin . During A number of transgenic macrophage reporter mice have proven particularly useful for studying macrophage biology , and havAnother layer of complexity lies in the pronounced phenotypic and functional diversity of macrophage subsets. Despite their developmental similarities, the function of macrophages in different organs varies. Red pulp macrophages, for example, are responsible for red blood cell clearance , while tWithin the skin, macrophages have long been thought to be a homogeneous group, primarily serving as a first line of defense against potentially invading pathogens. However, it is likely that skin-resident macrophages are more heterogeneous than previously appreciated. Certainly, macrophages within the skin exhibit a diverse distribution anatomically, where they may be associated with blood vessels , lymphatic vessels, or may reside in the intervascular space , 35. It Dendritic cells (DCs) represent a subpopulation of antigen-presenting cells that are largely defined by their capacity to present antigens to naïve T cells, for both the generation of immunity against invading pathogens, as well as tolerance to self-antigens and commensal bacteria . With reLangerhans cells (LC) reside in the basal and suprabasal epidermis, where they form a network between keratinocytes. Intravital experiments have shown that LCs remain sessile during steady-state conditions, but that their dendrites extend between keratinocytes in a repetitive fashion –45, whicDevelopmentally, LCs have more in common with macrophages than dermal dendritic cells. LCs develop from a primitive macrophage population during embryogenesis, which can be yolk-sac or fetal liver-derived ••, and i+ dermal dendritic cells, which further confounded interpretations of the LC-depletion models [Despite being discovered over 100 years ago, the exact immunological role of LCs remains controversial , 49. Earn models –65. Moren models , 67. Remn models –71, unden models –75.hiCD103-), but there are also CD11bloCD103+Langerin+ and CD11b-CD103- subsets [Dermal dendritic cells (DDCs), in contrast to LCs, are a more heterogenous population that relies upon continuous resupply from bone marrow-derived progenitors. In both mice and men, the dermis contains multiple DDC subsets, although the functional specialization of each subset remains largely unknown. In mice, the majority of DDCs express CD11b but not CD103 , based on their location and morphology . DETCs a−/− mice, which has been predicated upon the assumption that these cells represented the sole γδ T cell population within the skin. The recent discovery of an additional population of skin-resident γδ T cells, namely the dermal γδ T cells, necessitates a revision of these studies. For instance, while the increased keratinocyte apoptosis and delayed wound healing observed in TCRδ−/− mice is consistent with the role for DETCs in the epidermis [Staphylococcus aureus may be attributed to functions shared by dermal γδ T cells, in particular IL-17 production [DETCs form tight associations with E-cadherin expressed on keratinocytes , and thipidermis –114, theoduction .− γδ T cells, both within the dermis and epidermis, with only a few DETC. These γδ T cells could be elicited from IL-7-stimulated skin organ culture, suggesting they were normally skin-resident. More recently, we and others have extensively characterized this dermal γδ T cell subset [Despite the long-held view that DETCs were the sole γδ T cell population within the mouse skin, previous studies had indicated the presence of an additional skin-resident population of γδ T cells that do not express Vγ5 , 117. Inl subset , 119, coDermal γδ T cells constitute 50 % of the total dermal T cell population in mice, 30–50 % of which express the Vγ4 TCR. They are round or amoeboid in morphology, and a significant proportion are migratory •, albeit+ cells. Whether this population is the human equivalent to the murine dermal γδ T cell remains unclear, although the evidence to date suggests that the two populations do not equate. In contrast to murine γδ T cells, which are pre-committed to IL-17 production in the embryonic thymus [+ T cell lines produce TNFα and IFNγ when stimulated in culture [+Vδ2+ subset expressing cutaneous lymphocyte-associated antigen (CLA), the skin-homing receptor. Importantly, these circulating Vγ9+Vδ2+ cells produced IL-17 after bacterial infection and have been identified in psoriatic lesions [Human skin also contains a population of dermal γδ T cells, most of which express Vδ1 TCR, contrasting with the γδ T cells found in human peripheral blood that comprise largely Vδ2c thymus , 128, hu culture , 130. Hu lesions •. UndersInnate lymphoid cells (ILCs) are a family of newly described cells derived from a common lymphoid progenitor, and are identified by their lack of lineage marker expression  and their lymphoid morphology . They ar+ cells isolated from murine skin. Like other ILCs, dermal ILC2s required IL-7 for their development and survival, and could be replenished by bone marrow-derived cells following irradiation. Dermal ILC2s constitutively produced IL-13, and could upregulate IL-5 and IL-13 production when activated by systemic IL-2 treatment [Although originally described in mucosal tissues, significant numbers of ILC2s have been identified in the dermis of both mice and humans ••, 134••reatment ••, or byreatment ••. InterAlthough there is no evidence yet to date that ILC1s or ILC2s reside within normal skin, ILC3s were recruited to imiquimod-induced psoriasiform lesions •. WhetheResearch over the past few years has shed new light on the complex functions of skin-resident immune cells in homeostasis and inflammation. The discovery of novel cell populations, such as dermal γδ T cells and ILC2s, has expanded our knowledge of innate immune sensing and early responses towards pathogens entering the skin. How exactly epidermal and dermal inhabitants interact with each other and the environment and coordinate downstream adaptive immunity is still largely unexplored. The development of transgenic reporter mice with fluorescently tagged immune cell subsets in combination with advanced imaging approaches provides a unique opportunity for furthering our understanding of cutaneous biology in the steady-state and during disease conditions in the years to come.


Linguistic evolution mirrors cultural evolution, of which one of the most decisive steps was the "agricultural revolution" that occurred 11,000 years ago in W. Asia. Traditional comparative historical linguistics becomes inaccurate for time depths greater than, say, 10 kyr. Therefore it is difficult to determine whether decisive events in human prehistory have had an observable impact on human language. Here we supplement the traditional methodology with independent statistical measures showing that following the transition to agriculture, languages of W. Asia underwent a transition from biconsonantal (2c) to triconsonantal (3c) morphology. Two independent proofs for this are provided. Firstly the reconstructed Proto-Semitic fire and hunting lexicons are predominantly 2c, whereas the farming lexicon is almost exclusively 3c in structure. Secondly, while Biblical verbs show the usual Zipf exponent of about 1, their 2c subset exhibits a larger exponent. After the 2c > 3c transition, this could arise from a faster decay in the frequency of use of the less common 2c verbs. Using an established frequency-dependent word replacement rate, we calculate that the observed increase in the Zipf exponent has occurred over the 7,500 years predating Biblical Hebrew namely, starting with the transition to agriculture. Between 15,000 and 10,000 years ago, a major transition in human sustenance was instigated in W. Asia, which set the stage for modern human society: the transition to agriculture . The high frequency words obeying the original Zipf law constitute a "kernel lexicon", whereas the vast low frequency part consists of more specific terminology.The Zipf exponent α is about 1 in natural languages, while l corpus . A simill corpus . Eq. (1)k(f), that is larger for less frequently used words [k(f), their Zipf exponent would increase with time. The fact that α is always about 1 suggests that new words are formed with a similar k(f), and these balance the death of the old ones.  Word frequency is intimately connected to language history. It was already noted e.g., in Chap. 3 of Zipf's book , that hied words ,29. If wk(f) determined by Pagel et al. [ It follows that for any non-productive, morphologically distinguishable lexical subset (consisting of word types that stopped being created), α should increase with time. Here we analyze the frequency of verbal roots in an ancient Semitic text, the Hebrew Bible. We find that while the total verbal corpus shows the expected Zipf behavior, the 2c verbal roots  exhibit l et al. , the incl et al. -32, and DAE). Borrowing is excluded based on expert opinions from the linguistic literature. Details and discussions of all reconstructions are given in the EA compiled by Yigal Bloch , the "hollow" II-w roots , and those with a reduplicated last consonant. By direct counting in a Biblical Concordance we find that this group corresponds to about 12% of BH nouns. These possibly originate from an early phase of the 2c language.  Next, we suggest two definitions of 2c roots. The narrow definition includes only strictly 2c nouns, such as *n and I-w plus most I-y and III-w/y roots [y and III-w/y roots . We avoid most plant and animal names that could have been introduced either before or after their domestication. This leaves mainly terms related to fire and hunting, whose inclusion in the lexicon is justified by archeological data discussed below. "The manufacture of stone tools and the manipulation of fire are the most important extrasomatic milestones in our early evolutionary trajectory" . "Fire pa priori probability for such 2c nouns in Semitic languages, say 12% in BH. It could be explained if most of the fire synonyms originate from an older proto-language that had an abundance of 2c lexemes. Normally, words are replaced approximately every 3,000 years, but some survive considerably longer [ The multifunctional role of fire is manifested by several PS synonyms for the noun “fire” . We can y longer ,29. ThesND [RPN [ Corroboration of our conclusions can indeed be obtained from older proto-languages. Preceding PS on the linguistic genealogical tree is PAA , from which the Afroasiatic (AA) language families  have evolved. Unlike Semitic and Old Egyptian, the other African languages have only recently been documented. Consequently, there is yet no consensus over the PAA lexicon, and whether it originated before or immediately after the transition to agriculture, i.e. between 9,000  to 12,00ND  and RPN ND [RPN . It encoND [RPN .nabl) does not. Indeed, “fire” was found to be one of the "ultraconserved words" in the Nostratic macrofamily [ Because our starting point is PS, we do not require that every item in these dictionaries be correct, only that they are sufficiently comprehensive to include the predecessors of most PS lexemes. The last two columns in rofamily . Thus alA sample space of 5 items might be too small for statistical inference. Hence we add the 10 synonyms for the verb “set afire, burn” in māy, whereas drinking (water) is depicted by the two PS/2c verbal roots *šty and *šqy [DAE entries 999, 1878 and 1209), hence of pre-agricultural origin.like fire, is one of the "bare essentials" required to sustain life. Thus water vocabulary should also be immune to replacement. The PS noun for “water” is 2c *nd *šqy . All thrbow (#2.3) with which to shoot (#2.4) an arrow (#2.1), a small bag (#2.2) for collected items, perhaps a water bottle. All these PS terms are 2c and all are attested in PAA. The prehistory of the bow is difficult to determine because most components (except arrowheads) are perishable. Ballistic arrowhead analysis concluded [qaš-t, “bow”, is agreeably PAA, and there is no obvious linguistic trace for “atlatl”, suggests that Natufians have utilized predominantly bows and arrows and/or that the term for atlatl has undergone a semantic shift to indicate the bow.oncluded  that litoncluded ,38. The ṣd (#2.5). While in BH it means “to hunt”, in Akk. duṣâ means “to prowl, turn about” [turn around, completing an elliptic annual trajectory.In addition to the overwhelming 2c vs. 3c statistics, we note an interesting polysemy (multiple meanings) of the PS root *n about” . Prowlinn about” . Hunter-provisions (#2.6). In PS, these must have been considered as homonyms (see EA), because for the farmer there was no connection between “hunting” and “provisions” (the latter coming mainly from his domesticated fauna and flora). For his hunter-gatherer predecessor, however, these must have been strongly associated, because provisions carried on hunting journeys might have included dried/smoked meat of hunted animals. Such a connection between hunting, prowling and provisions is thus indicative of a nomadic hunter-gatherer society. The pre-agricultural origin of this verb is supported by a possible pre-agricultural connection between hunting (#2.5) and  Evidently, more farming than hunting terms survived in PS, and nearly all have 3c morphology. farmer (#3.1) lived in a large village (#3.15), constructed of square houses [straw (#3.26) reinforced [bricks (#3.17). Indeed, straw became readily available after the Pre-Pottery Neolithic A (PPNA) wheat domestication [ As of the Pre-Pottery Neolithic B (PPNB), ca. 10,500 BP, the farmer #.1 lived inforced  sun-bakefield , which he would plow (#3.12), sow (#3.27) or plant (#3.25). Furrow tracts from W. Europe date to 5,500 BP [canal (#3.19), over 7 kyr old, was discovered in Choga Mami, 110 km E. of Baghdad: "It is conceivable, indeed probable, that plough cultivation accompanies irrigation agriculture in the earlier Samarra period" [ The farmer would work in an agricultural 5,500 BP  and must5,500 BP . This ma period" . granaries (#3.2 and 3.9), about 11,300 years old, were unearthed in the Dead-Sea region near Dhraʽ, Jordan [ Tilled fields can be sown only if grain from the previous year is stored under adequate conditions. PPNA , Jordan . These rwells (#3.4) were dug by Neolithic farmers on the coast of Cyprus ca. 9,200 BP [ˀ morphology of *biˀr “well” (#3.4) is also attested in the PS/3c verbal root for drawing water , possibly because they have originated in the same period. The first attested 9,200 BP . The old9,200 BP ,46. A Po9,200 BP . Thus wecooking (#3.5 and 3.24), leading to a "culinary revolution" nearly 9,000 years ago [ The earliest mineral-tempered ceramics from Tell Sabi Abyad (N. Syria) was likely introduced for ears ago , when (Pears ago  and “roaLivestock (#3.8) was often composed of mixed sheep and goat herds that optimize vegetation exploitation. This contrasts with their non-overlapping habitats in Nature [ˀn “livestock” (#3.8) is a post-agricultural innovation. The herd was lead by a stockbreeder (#3.18) to a drinking trough (#3.21). Due to lactose intolerance, milk utilization has begun rather late, at the end of the Chalcolithic or the Early Bronze [butter (#3.14), a low lactose milk product. Herding began after goat and sheep domestication, either in the Neolithic or as late as the Chalcolithic . Liveston Nature , suggesty Bronze . Howevery Bronze . In agreFermenting wine (#3.13) was made from grapes (#3.3) already in the Neolithic: jars from Georgia (in the Caucasus), dating to ca. 8,000 BP, were shown to contain resinated wine deposits, as have 7,300 BP sherds from the Zagros Mountains in Iran ([terebinth tree (#3.6), Pistacia atlantica [karm, *karān “vineyard” (#3.16): Mt. Karmel in N. Israel and Karānā in Upper Mesopotamia . Although a dry wasteland today, the high concentration of archeological mounds suggests it has once been fertile land .Beer (#3.23a) was the most popular intoxicating (#3.23b) drink in Mesopotamia. Until recently, the earliest evidence for beer  was found in the Sumerian trading post of Godin Tepe in Iran [ in Iran . But rec in Iran . Millet (#3.7) was domesticated in N.E. China about 10,000 years ago [ears ago . It madeears ago , just inExceptions to the regularity demonstrated in marr “a hoe”, derived from the 2c root *mrr “to hoe”, originates within a PAA farming lexicon [ND #1482 [mrr has itself been borrowed by Sumerian from PAA [herding, *rˁy, is 2c although herding postdates ungulate domestication that occurred after the transition to agriculture. However, in some Chadic dialects it means “to chase, follow”, DAE #663 [ lexicon . The nouND #1482 , or a "wND #1482 . If the from PAA , then itfrom PAA , an actiDAE #663 . This maDAE #663 , includiDAE #663 . Thus iff(r), for the BH (non-Aramaic) verbal roots with f ≥ 10 [α0=1.07and α1≈2 (dashed lines). The switchover occurs around f = 20, so that the kernel of BH is characterized by f ≥ 20.  The study thus far focused on statistics of culturally specific terms (hunting vs. farming) that could be correlated with archeology. These are mostly low frequency lexemes, hence not belonging to the "kernel lexicon". We now analyze the kernel lexicon of an Ancient Semitic text, the Hebrew Bible, bisecting it into its 2c vs. 3c components. We consider verbs, because their 2c vs. 3c origin can be determined rather mechanically (see Methods), allowing processing a large number of verbs. Yet they constitute the only part of speech whose Zipf plot is similar to that of the whole corpus . The blah f ≥ 10 . It indeh f ≥ 10 , with αmeanings) are replaced (by other words of the same meaning) less often than the less frequent ones. Thus if the 2c stratum indeed predated the 3c one, the frequently used 2c lexemes may have simply survived replacement during the subsequent 3c era. This is supported by their frequency-rank dependence in α2c=1.28), whereas the high frequency 3c verbs have a smaller α3c=0.82(linear fits not shown). This might be explainable by the 2c > 3c transition: while the 2c language was alive, 2c words of a given meaning were depleted at the same rate as alternate 2c lexemes were generated, and the language maintained its steady-state with the usual exponent α ≈ 1. After the 2c era has ended, 2c roots were no longer created only eliminated. Because less frequently used words decay faster, α2c increased with time.  As recently shown , frequent = 0) they started to be replaced with new 3c words. Assume that (up to a constant) the frequency of use of a certain verbal meaning (at least in the kernel lexicon),f0(r) , is an inherent property of human language and hence not strongly time-dependent. We thus equate it with the frequency of the total verb distribution =A2cf0(r), where A2c is some constant. We expect A2c>1 if the 2c corpus was once used more frequently than today . One may turn this into a quantitative method for dating the 2c > 3c transition. Suppose that once there were only 2c words, and at some time  , is a unique function of the (time-independent) verb meaning frequency,f0(r). Eventually, after some time t that we opt to determine,f2c reached the values observed in the Biblical lexicon  is required. We adopt Pagel et al. [ To proceed, a functional form for l et al.  power-lal et al. :B = 0.55 kyr-1 and β = 0.13. We do not vary these parameters in fitting our data. However, in Ref. [From the correlation line for English verbs in their Figure 3a, one estimates  in Ref.  the freqt and A2c to fit the 2c data (blue triangles), we obtain the blue line agreeing with the data over the whole frequency range, even where it deviates from Zipf's law, t = 7.8 kyr. Adding the presumed age of BH, ca. 3 kyr, gives 10.8 kyr for the 2c > 3c transition, agreeing nicely with the onset of agriculture. Adjusting growth rather than decay: An analogous model may describe the 3c verbs, which experience exponential same parameters as above (excepting A3c) we obtain the red line in Of course, such growth cannot go on indefinitely, but we assume the time-depth is not large enough to observe saturation. With exactly the f0), or the full line (when the circles are used asf0). Thus the "burning verbs" behave like the entire BH verb population, both yielding the same date for the 2c > 3c transition. As a check for the robustness of this analysis, we return to the "burning verbs" discussed in Subsec. I(b) above. We find 10 such verbs in BH  Most of them are also PAA and/or Nostratic  and (b) of 2c morphology. A similar trend is observed for PS hunting terms, which are all 2c.all have 3c morphology, and only rarely possess PAA cognates. We were able to find very few exceptions to this rule, and these represent secondary use of existing 2c roots. Hence PS hunting vs. farming lexicons have, on average, different time-depths and morphologies. Likely, then, a 2c-enriched hunter-gatherer language has evolved into a 3c-dominated farmer language with the transition to agriculture in W. Asia.  The farming terms collected in  This suggestion is corroborated by a frequency analysis of BH verbs. While the total verbal corpus exhibits a Zipf plot with the expected exponent of about unity, its 2c subset has an observably larger exponent. This can be understood if the creation of new 2c roots ceased sometime in prehistory, and thereafter the use of the low frequency 2c verbs decreased faster than those of higher frequencies. This was turned into a novel quantitative method for dating the 2c > 3c transition. The date obtained, nearly 11 kyr BP, indeed marks the transition from hunting to farming. Thus two independent methods, applied to different parts of the Semitic lexicon ("specific" vs. "kernel"), lead to the same conclusion namely, that a major change in human lifestyle (the transition to agriculture) correlates, in W. Asia, with a major linguistic change.Text S1Explains how the Proto-Semitic word reconstruction was achieved.(PDF)Click here for additional data file.Table S1Etymological Appendix for (PDF)Click here for additional data file.Table S2Etymological Appendix for (PDF)Click here for additional data file.Table S3Etymological Appendix for (PDF)Click here for additional data file.Table S4Etymological Appendix for (PDF)Click here for additional data file.Text S2List of references for the Etymological Appendix.(PDF)Click here for additional data file.Text S3The complete Etymological Appendix composed of the above six supporting files.(PDF)Click here for additional data file.Table S5Ranking of “weak”  verbal roots in the Hebrew Bible by frequency. The 2c roots were extracted from the list in Ref. 31 according to the "broad definition" in the Methods section. (PDF)Click here for additional data file.Table S6Transliteration: Proto-Semitic consonant phonemes with their Hebrew and Arabic equivalents.(PDF)Click here for additional data file.