In adults, there are differences between male and female structural and functional brain connectivity, specifically for those regions involved in pain processing. This may partly explain the observed sex differences in pain sensitivity, tolerance, and inhibitory control, and in the development of chronic pain. However, it is not known if these differences exist from birth. Cortical activity in response to a painful stimulus can be observed in the human neonatal brain, but this nociceptive activity continues to develop in the postnatal period and is qualitatively different from that of adults, partly due to the considerable cortical maturation during this time. This research aimed to investigate the effects of sex and prematurity on the magnitude and spatial distribution pattern of the long-latency nociceptive event-related potential (nERP) using electroencephalography (EEG). We measured the cortical response time-locked to a clinically required heel lance in 81 neonates born between 29 and 42 weeks gestational age . The results show that heel lance results in a spatially widespread nERP response in the majority of newborns. Importantly, a widespread pattern is significantly more likely to occur in females, irrespective of gestational age at birth. This effect is not observed for the short latency somatosensory waveform in the same infants, indicating that it is selective for the nociceptive component of the response. These results suggest the early onset of a greater anatomical and functional connectivity reported in the adult female brain, and indicate the presence of pain-related sex differences from birth. •Noxious stimulation causes widespread pain related potentials in the neonatal brain.•This widespread pain response is more likely to occur in female babies.•Brain responses to touch do not differ between male and female babies.•Sex differences in human brain pain pathways are present from birth. Although immature, the neonatal nociceptive system has the potential to encode painful stimuli . NocicepIn adults, pain processing differs between males and females. Following experimental noxious stimulation, female subjects have higher sensitivity, lower tolerance, and reduced endogenous inhibitory control . Recent It remains unclear whether these sex differences in pain processing are present from birth, or whether they emerge later in life. Previous studies have reported conflicting results with respect to sex differences in human infant pain behaviour following acute noxious stimulation , but it Since (i) some of the structural and functional brain networks related to pain processing are sexually dimorphic in adults and (ii) those same networks undergo significant development in the third trimester and postnatal period e.g. , we hypoParticipants were 81 newborn infants, no older than 2 weeks postnatal age . The noxious stimulus was a clinically required heel lance used for blood sampling performed by an experienced research nurse and time-locked to the ongoing EEG recording. A more detailed description of the lance procedure and EEG protocol has been published elsewhere .Recording electrodes (Ambu Neuroline disposable Ag/AgCl cup electrodes) were positioned individually by a clinical physiologist according to a modified international 10/10 electrode placement system at Fp1, Fp2, F3, F4, F7, F8, FCz, Cz, C3, C4, T7, T8, CPz, CP3, CP4, TP9, TP10, P7, P8, POz, O1, and O2. Although it was not always possible to apply the full set of electrodes, in 91% of the neonates at least 11 electrodes were used, and the Cz electrode (midline vertex) was used in all recordings . The refEEG traces were analysed using EEGLAB 10.2.5.8b , which is a negative-positive waveform typically occurring within 400–700 ms post-stimulus (N3P3) . As a wiThe incidence, amplitude, and scalp distribution of both waveforms, including the scalp location of the maximal amplitude, were recorded. Since both the N2P2 and N3P3 have been defined previously at vertex channels ; 2010b, 2 omnibus test) and linear  regressions (using the ‘enter’ method). The same analysis was then repeated using gestational age at birth as the predictor variable. As EEG response amplitude is inversely related to the distance of the source from the scalp, it is plausible that a smaller response amplitude might be detected in infants with a larger head size as a result of a greater scalp thickness and distance from the source. Similarly, it may be more likely to detect an EEG response at a greater number of scalp locations in infants with a smaller head size. Therefore, for regressions including response amplitude and distribution we controlled for this using head circumference (measured at birth), which was correlated with GA (r (81) = 0.83, p < 0.001). The strong correlation between GA and head circumference (HC) at birth resulted in a degree of multicollinearity ; therefore these regressions were re-run without HC to confirm that similar results were obtained. Similarly, for the distribution analysis, the maximal response amplitude was added as an additional confounding factor. Odds ratios (Exp(b) values) are reported for logistic regressions; these represent the increased likelihood of belonging to a test category vs. a reference category , with every increase in the predictor (e.g. a 1-week increase in GA). A summary of the sample sizes for these analyses is presented in Statistical analysis was conducted in SPSS (IBM Corp.). The effect of sex on the incidence, amplitude and distribution (focused at the vertex/widespread2 (2) = 1.00, p = .606) were filled in using the maximisation estimation method  = .001, p = .981; amplitude: F  = 1.42, p = .250, OR = .14, p = .263). Similarly, the incidence and amplitude of the N3P3 response at the vertex were not significantly related to the gestational age at birth of the subjects (incidence: χ2 (1) = .35, p = .552; amplitude: F  = .90, p = .412, OR = .11, p = .611), and clear responses could be identified even in the youngest babies  in 85.2% (69/81) of the trials. t babies . Removint babies .Fig. 2Av2 (2) = .52, p = .773). There was a trend for the location of the maximal response to depend on the gestational age at birth of the subjects, as younger infants were less likely to have a peak at the vertex and peri-central electrodes compared to older infants (vertex vs “elsewhere”: Odds ratio (OR) = 1.28,2 (2) = 5.19, p = .075). The amplitude of the maximal response was not related to the sex or age of the subjects  = 2.65, p = .078, OR = .13, p = .290; age: F  = 2.23, p = .116, OR = −.12, p = .561). Removing HC at birth from the model resulted in a significant relationship between GA and maximal amplitude (2 (2) = 14.36, p = .001, vertex vs “elsewhere”: OR = 1.51, p = .001; peri-central vs. “elsewhere”: OR = 1.26, p = .145).Although a nERP could be identified at Cz or CPz in 85% of trials, this response at the vertex was not always the maximal response. The maximal N3P3 response was located at the vertex electrodes in 73.9% (51/69) of trials, at the peri-central electrodes in 8.7% , and elsewhere in 17.4% (12/69). Sex was not a predictor of the location of the maximal response  = 13.61, p = .003, OR = 7.52, p = .004; 2 (3) = 5.67, p = .129, OR = .76, p = .149). Removing HC from this analysis confirmed that there was still no effect of GA on the response distribution (2 (3) = 3.09, p = .378, OR = .67, p = .569; age: χ2 (3) = 5.01, p = .171, OR = .71, p = .133).We next explored the spatial distribution of the nERP across the scalp. We found that although the nERP could be identified at the vertex in 85% of trials and was maximal at these electrodes in 74% of trials, it was not always focused at this location see . This waribution . In contIn light of the effects of age and sex on the position of the maximal response and distribution pattern, respectively, we also investigated any age and sex interaction on the maximal response position and on the distribution pattern using separate regressions for males and females, with GA as a predictor. We did not observe an interaction between the dependent variables.Here we have shown that there are already significant sex differences in the distribution of nociceptive cortical event related activity in newborn infants, both preterm and full-term.Most neonates had a maximal nERP at the vertex, but this is less likely for neonates born at a younger gestational age. Moreover, we found that this maximal response is not always narrowly focused at the vertex, but rather 64% of infants exhibit a significant response across several electrodes. Importantly, females are more likely to exhibit such a widespread pattern compared to males, independent of their gestational age at birth or head size, an effect which is selective for the nERP, the nociceptive component of the ERP response. With limited data on sex differences in the neonatal literature, this finding provides a significant contribution to the ongoing debate about the mechanisms underlying sex differences and pain .The fact that the nociceptive response was widespread in most of the subjects has not been reported previously as earlier infant nERP analyses have been restricted to the vertex response . The N3PFemales are more likely to have a widespread response than males. This may reflect the greater overall anatomical and functional connectivity and more efficient network organisation reported in the adult female brain . As wellThe greater activation and connectivity in female brains, both overall and following noxious stimulation, may underlie the differences between males and females in pain perception and self-report. Females are more sensitive to certain noxious stimuli , or exhiPrevious behavioural studies have produced conflicting results for the existence of pain-related sex differences in neonates. Following a noxious stimulus, Grunau et al. found that males have shorter latency time to cry and onset of facial changes compared to females , while GPrevious research indicates that noxious heel lance elicits a response that is maximal at the vertex , 2010c. Potentials that peak at midline electrodes are associated with bilateral cortical sources, with activation in both hemispheres summating at the midline . This isSome trials exhibited a peak response at peri-central electrodes rather than the midline vertex. It is possible that these non-topographically organised responses reflect normal individual variability, particularly as many cortical structures are still maturing . AlternaThe incidence of a vertex response was not associated with gestational age at birth in the present study. Previous research from our own group and others has reported an age-dependent shift from non-specific delta brush responses to nociceptive-specific event-related potentials , with veTwo methodological considerations deserve mention. In the present study, we chose to control for head circumference at birth when assessing the effect of gestational age at birth on the amplitude and distribution of the responses, because we assumed that they might be modulated by head size. Unsurprisingly, GA and HC at birth were highly correlated, resulting in a degree of multicollinearity. Removing HC confirmed that results were consistent, except for the analysis of maximal response amplitude. Both GA and HC significantly predicted the amplitude of the maximal response across the scalp when tested on their own, but not when tested together. This highlights the importance of considering head circumference as a confounding factor when assessing developmental effects on response amplitude.EEG studies exploring responses to sensory stimuli typically involve multiple repetitions per participant, allowing averaging within participants to increase the signal-to-noise ratio. This is not feasible in studies assessing cortical responses to noxious tissue-damaging stimuli in neonates, and the present results are necessarily based on single-trial data. However, noxious stimuli elicit large-amplitude responses in neonates (e.g. mean 59.2 ± 26.3 μV at the vertex in the present study) that are clearly discernible relative to the ongoing EEG within single trials. Furthermore, previous research has indicated that these are reproducible in babies requiring multiple procedures , and conWhile neonatal nociceptive event related potentials elicited by a clinically required heel lance are often maximal at the vertex, the overall nERP response is more likely to be widespread across the scalp, rather than focused at the vertex. Moreover, females are more likely to exhibit a widespread response, independent of gestational age at birth. This may reflect greater activation or connectivity in female brains following noxious stimulation, from early development. As female adults are more sensitive to experimental pain, and are more likely to develop clinical pain conditions, this novel finding may reflect the presence of pain-related sex differences from birth.Conceptualization of study: M.F., L.F., J.M., M.V.; experimental design: M.F., L.F., M.V., J.M.; data collection: K.W., M.L-D., L.J., M.V.; data analysis: L.J., M.V., H.P., I.P.; data interpretation: L.J., M.V., L.F., M.F., J.M., K.W.; manuscript preparation: L.J., M.V.; discussion of results, critical comments and revision of manuscript: L.J., M.V., L.F., M.F., J.M., M.L-D., K.W.


Capsicum genotypes. Placentas of Habanero peppers can incorporate inorganic nitrogen into amino acids and have the ability to secure the availability of the required amino acids for CAP biosynthesis. In order to determine the participation of the placental tissue as a supplier of these amino acids, the effects of blocking the synthesis of Val and Phe by using specific enzyme inhibitors were analyzed. Isolated placentas maintained in vitro were used to rule out external sources′ participation. Blocking Phe synthesis, through the inhibition of arogenate dehydratase, significantly decreased CAP accumulation suggesting that at least part of Phe required in this process has to be produced in situ. Chlorsulfuron inhibition of acetolactate synthase, involved in Val synthesis, decreased not only Val accumulation but also that of CAP, pointing out that the requirement for this amino acid can also be fulfilled by this tissue. The presented data demonstrates that CAP accumulation in in vitro maintained placentas can be accomplished through the in situ availability of Val and Phe and suggests that the synthesis of the fatty acid chain moiety may be a limiting factor in the biosynthesis of these alkaloids.Capsaicinoids (CAP) are nitrogenous metabolites formed from valine  and phenylalanine (Phe) in the placentas of hot  Capsicum spp.) have long been valued worldwide as an important source of bioactive compounds that are beneficial to human health. Their fruits are known not only as an important source of vitamins, carotenoids and phenolic compounds, all of which possess important antioxidant properties, but also for their pungency [Chili plants . The ability to synthesize CAP is related to the presence of at least one functional acyltransferase allele (AT3) at the Pun1 locus [Pun1  cycle [in vitro.Capsaicinoids (CAP), the pungent principle of peppers, are exclusively produced and accumulated in placentas of the hot genotypes of peppers , the balocus C; , and evelocus C; . This sulocus C; . Moreovelocus C; ,10. InteT) cycle . In this−1 FW; [−1 FW; [C. chinense placentas are very low, suggesting that their synthesis must be triggered only when it is required to satisfy specific demands [It has been demonstrated that both amino acids have been shown to be required to increase CAP synthesis in non-differentiated cell cultures ,13. Howe−1 FW; ) and Val [−1 FW; ) cell po demands .−1·mg−1 protein, with pericarps displaying the highest activities, about 2-fold higher than that found in placentas , with pericarps and placentas presenting much higher activities over leaves (Acetolactate synthase (ALS) represents the first committed step in the synthesis of Val, whereas arogenate dehydratase (ADT), directly channels arogenate towards Phe formation . Both enlacentas A. ALS acr leaves B. In botin vitro  and ALS , respectively. Maximal ADT inhibition (86%) was attained using 500 μM of p-FP for 72 h on placentas cultured in vitro A. Maximain vitro B. Higherin vitro . Furtherin vitro .ca. 50% decrease in Phe contents , which suggests that Val synthesis could be a limiting factor in this process. The fact that by inhibiting Val synthesis, Phe content diminishes concomitantly, but that the inhibition of Phe does not affect Val content, suggests that Phe synthesis might be subordinated to the synthesis or availability of Val in placentas. Free amino acid pools are strictly controlled in plant cells and the content of one amino acid is kept in a precise balance with the others. Frequently, the internal concentration of one amino acid might control to some extent the synthesis of others, including those not biosynthetically related [Variations in soil nitrate availability affected its own contents in placentas and directly impacted CAP contents . This su sources . MoreoveAT cycle . Our resAT cycle  and Phe AT cycle  availabiAT cycle . The use related .in vitro system to maintain isolated placentas has resulted in a valuable asset, since environmental conditions can be strictly monitored, so the effects due to one sole factor can be observed [It has been proposed that in order to increase the production of secondary metabolites, the interface between primary and secondary metabolism could be genetically manipulated , so thisobserved  and the observed ,35.in vitro maintained placentas can be accomplished through in situ availability of Val and Phe and suggests that the synthesis of the fatty acid chain from Val may be a limiting factor in these alkaloids’ biosynthesis.Summarizing, our data demonstrates that CAP accumulation in C. chinense Jacq.) var. Chak k’an-iik were cultivated in a greenhouse at the Centro de Investigación Científica de Yucatán . Plants were grown over a mixture of soil, perlite, vermiculite, peat moss and coconut fiber, and watered to full field capacity every other day. They were fertilized once a week with MaxiGro® y MaxiBloon® . Unripened fruits were collected from the same plant stratum , washed with commercial soap and thoroughly rinsed in running tap water. Green pods  were collected approximately 25 days post anthesis (DPA). Placental in vitro system was established using five whole, isolated placentas (approx. 3 g−1 FW) maintained for the duration of the experiments in Erlenmeyer flasks containing 40 mL of MS medium [−1 sucrose without growth regulators. Flasks were maintained at 25 °C, with continuous light and gentle shaking (80 rpm) for the duration of the inhibition experiments.Habanero pepper plants (S medium  supplemein vitro system described above. Cln was dissolved in tetrahydrofuran (THF) to a concentration of 12 μM and diluted in 5 mM K2HPO4 (pH 7.5), whereas p-FP was dissolved in water. Both were filter-sterilized prior to application. Samples were collected, as indicated in figures, frozen with liquid nitrogen and stored at −80 °C until analysis. Control flasks were subjected to the same manipulations, except that THF, phosphate buffer or water were added instead of the inhibitors. Samples were used to determine ADT and ALS specific activities and mRNA’s levels, as well as Phe, Val and CAP contents.Phenylalanine and valine synthesis were blocked using p-FP and Cln, respectively. Inhibitor treatments (time and dose) were experimentally determined, applying up to 500 μM during 24 and 48 h for p-FP  and up tp/v) polyvinylpolypyrrolidone) in a 1:2 (p/v) ratio. The slurry was homogenized with a Polytron for 2 min and then, centrifuged at 20,400× g for 30 min. All manipulations were performed at 4 °C. Total protein extracts were used in the enzymatic assays. ALS activity was measured in a reaction mixture containing 20 mM phosphate buffer pH 7.0, 20 mM pyruvate, 0.5 mM thiamine, 0.5 mM MgCl2, 10 µM FAD+ and 1–5 µg total protein [2SO4 and then quantified at 525 nm, in the presence of 500 µL 0.5% creatine and 500 µL 5% α-naphtol, prepared in 2.5 N NaOH. In order to estimate ADT activity, arogenate had to be enzymatically prepared from prephenate [C. chinense leaves. ADT activity was measured in a reaction mixture containing 50 mM Tris-HCl (pH 7.5), 2.5 mM arogenate, 0.25 mM Tyr, 1 µM leupeptin, 1 µM pepstatin, and 0.25–1 µg total protein in a final volume of 200 µL [v/v), flux: 1 mL/min and running time: 6 min. Derivative reaction products were detected at 360 nm (excitation) and 455 nm (emission). The range for the Phe standard curve was 0–100 nmol. ALS activity was defined as nmol acetoin min−1·mg−1 protein, whereas ADT activity was expressed as nmol Phe min−1·mg−1 protein. Protein content was determined according to Peterson [Enzyme activities were assayed in protein extracts obtained from pulverized frozen placentas. Powder was mixed with the extraction buffer , 600 nM forward primer (0.6 μL), 600 nM reverse primer (0.6 μL), 75 ng RNA (~2 μL) and 6.8 μL RNAse-free water. RT-PCR conditions were 10 min, 95 °C, 40 cycles: (a) denaturation: 15 seg, 95 °C; (b) aligment/extension: 1 min, 60 °C  and (c) extension: 72 °C, 30 seg. Control samples were used to estimate differences in gene expression levels, according to the equation 2T−ΔΔC [Total RNA was extracted using the method described by Rubio-Piña and Vázquez-Flota . Primersm annuum . For ADTn 2−ΔΔCT .All of the data were subjected to analysis of variance (ANOVA), and mean comparisons were made using Tukey’s multiple-range test at a 5% level of probability.


The aim of this study was to explore the influence of workplace incivility on the emotional exhaustion of recreational sport/fitness club providers through a cross-level analysis. A total of 200 recreational sport/fitness club providers from Taiwan were selected for the repeated collection of measures and a 10-day diary method was used. The effect of workplace incivility on recreational sport/fitness club employees’ emotional exhaustion on a daily basis was analyzed at the intra-personal level, and the relationship between psychological capital and perceived service climate was studied at the inter-personal level. Five hypotheses were developed and tested using hierarchical linear modeling. The results found that employees’ emotional exhaustion and burnout highly correlated with workplace incivility and service climate. Based on the results, recommendations for employees and sport/fitness centers are proposed. Furthermore, research limitations and future directions are discussed. The market share of recreational sport/fitness clubs is consistently expanding and evolving. Over the years, private clubs, such as World Gym, BEING Sport, Curves, and Fitness Factory have become established in many different cities in Taiwan and continue to grow . AccordiThe diary method, as Hershcovis and Reich  pointed Employees at health and fitness centers are the key to service quality and customer satisfaction. However, the workplace incivility they face on a daily basis may be sabotaging their performance, and the negative effect of emotional exhaustion is largely overlooked. Therefore, the present study used the diary method to collect individual, intrapersonal perceptions of workplace incivility and emotional burnout, and analyzed the relationship between incivility and emotional burnout. In addition, the study also collected interpersonal data, such as psychological capital (PsyCap) and perceived service climate and performed a cross-sectional analysis to examine the sport/fitness clubs’ cross-level impacts on employee burnout.In an organizational context, affective event theory (AET) explains employees’ everyday mood and emotions . JohonsoHypothesis 1 (H1).Workplace incivility has positive effects on emotional burnout.PsyCap is characterized by four qualities: efficacy, optimism, hope and resilience ,14. As LHypothesis 2 (H2).PsyCap has negative effects on emotional burnout.Workplace incivility happens all the time. Interrupting phone calls, expressing a lack of respect, and unfair treatment are typical examples of workplace incivility. It produces negative emotions, resulting in employees’ no longer wanting to contribute to the organization . As PughHypothesis 3 (H3).PsyCap has a moderating effect on the relationship between workplace incivility and emotional exhaustion.The literature evidence suggested that PsyCap leads to positive moods, fosters personal well-being, increases organizational commitment, and improves job performance . SchneidHypothesis 4 (H4).Service climate has a significant positive effect on PsyCap.The job demands–resources model proposes that an organization’s service climate requires employees to engage with job resources. Their engagement not only influences their mental health, but also increases their job performance . SeveralHypothesis 5 (H5).Perceived service climate has significant negative effects on emotional burnout.The subjects of the research were members of the service staff at health and fitness centers located in different cities, including Taipei City, New Taipei City, Chiayi City, Chiayi County, Tainan City, and Kaohsiung City in Taiwan. A total of 2000 employees were selected to participate in the research. Data were collected using the 10-day diary method. In order to increase the response rate, a mobile text message was set to send to each individual participant at 17:00 every afternoon. All the subjects were informed about the data collection procedure when they were invited to participate in the research. Data were collected under a clearly informed consent agreement. The research was reviewed and approved by the Institutional Review Board. A total of 1790 valid responses were retrieved, with a valid return rate of 89.5%.Research participants were invited to complete their 10-day diary within a month. Workplace incivility and emotional burnout were assessed on a daily basis; therefore, data were retrieved for 10 consecutive days. On the other hand, PsyCap and service climate—which were individual level variables—were collected on the first day of participation and examined accordingly. The scales used in the research and demographic variables are presented as follows.Demographic variables were designed to meet the specific research needs. Demographic variables included gender , age, education level , marital status , perceived health status (ranging from 1 = “bad” to 5 = “excellent”), and length of service (in years).The workplace incivility checklist used in the present study was modified from Liu and Dai . The che2 / df = 3.33, GFI = 0.98, CFI = 0.98, NNFI = 0.95, RMSEA= 0.07). Cronbach’s α was 0.900, indicating that the reliability of the scale was acceptable.The Emotional Exhaustion Scale was modified from the measurement of experienced burnout originally introduced by Maslach and Jackson . Stateme2 / df = 3.17, GFI = 0.93, CFI = 0.96, NNFI = 0.94, RMSEA= 0.07). Cronbach’s α was 0.907, indicating that the reliability of the scale was acceptable.Psychological capital scale was adopted from Luthans, Youssef, and Avolio  and Yu e2/df = 5.16, GFI = 0.90, CFI = 0.96, NNFI = 0.93, RMSEA= 0.09). Cronbach’s α was 0.927, indicating that the reliability of the scale was acceptable.The perceived service climate scale was modified from Schneider et al. , which hThe control variables which were considered to affect employees’ psychology and behavior were gender, age, education level, health status and length of service. Control variables were selected based on relevant research ,33.Statistical analysis was performed using SPSS 21.0 for Windows   and HLM n = 1790), most of them were female (M = 1.68), married (M = 1.77), with a college degree (M = 3.04), at an age of around 30 years old (M = 30.42), and their average length of service was 5.29 years. As for perceived health status, most of them felt their health status was good . As for psychological capital and perceived service climate, most participants had high psychological capital (M = 3.46), and perceived high service climate (M = 3.88).Five research hypotheses were formulated and tested using hierarchical linear modeling. Results were presented as follows.2 = 1487.98, df = 178, p < 0.001), and that ICC (1) was 0.424, indicating that the individual factor accounted for 42.4% of variance of employees’ emotional burnout. According to James [The research intended to analyze the effects of within-subject variables and between-subject variables on the emotional exhaustion of frontline service employees at health and fitness centers. The cross-level analysis was primarily verified using null model analysis. Bliese  suggesteto James , an ICC to James  suggesteEmotional Exhaustion•Level-1 Model✓j = ψ0j + emjEmotional Exhaustion_ m•Level-2 Model✓0j = γ00 + u0jψ•Workplace incivility•Level-1 Model✓0j + emjWorkplace incivility_ mj = ψ•Level-2 Model✓0j = γ00 + u0jψ06 = −0.216). This indicated that perceived health status was negatively associated with emotional burnout.In 1ψ = 0.230), indicating that H1 was supported. PsyCap, however, had significant negative effects on emotional burnout (γ07 = −0.318); thus, H2 was supported. Furthermore, PsyCap was found to have no mediating effects on the relationship between incivility and burnout. H3 was rejected. Lastly, H5 was supported because perceived service climate had significant negative effects on burnout (γ08 = −0.118).Workplace incivility had significant positive effects on emotional exhaustion (•Level-1 Model✓0j + ψ1j* (Workplace incivility mj) + emjEmotional Exhaustion_mj = ψ•Level-2 Model✓0j = γ00 + γ01*(Gender_Mj) + γ02*(Age_Mj) + γ03*(Marrige_Mj) + γ04*(Eduation_Mj) + γ05*(length of service_Mj) + γ06* + γ07* (PsyCap l_Mj) + γ08*(Perceived service climate_Mj) + u0jψ✓1j = γ10+ γ11*(PsyCap _Mj)ψp < 0.05). Higher levels of PsyCap were associated with higher levels of perceived service climate. Results of hypothesis testing are presented in The results of regression reported the significant positive effect of PsyCap on the perceived service climate , perceived health status was highly associated with emotional exhaustion. Service providers who considered themselves as having a high perceived health status were less likely to experience emotional burnout. Similarly, health care professionals with higher physical health perception were found to have lower fatigue and stress . IndividThe 10-day diary method was utilized to collect repeated measures from service staff at health and fitness clubs. With five control variables controlled, research findings indicated that workplace incivility might cause employees’ emotional exhaustion. The findings were consistent with affective events theory (AET), that claims that workplace incivility might influence employees’ everyday emotion and behaviors . CortinaThree common causes of workplace incivility are abusive supervision, employee mistreatment or discrimination, and disrespectful customers ,49. The Throughout the cross-level analysis, the result indicated that PsyCap had no moderating effect on the relation between workplace incivility and emotional burnout. PsyCap, on the other hand, was found to have significant negative effects on emotional exhaustion, and so did perceived service climate. PsyCap, according to Luthans, Youssef, and Avolio , is compThe results showed that service climate had a negative impact on emotional burnout. An encouraging service climate plays a vital role in service quality . The finResults showed that service climate was found to be significantly positively related to PsyCap, which is consistent with Liu’s assertion  that posAccording to our findings, first, we found that among five control variables , perceived health status was positively correlated with emotional exhaustion. Second, we found that workplace incivility might cause employees’ emotional exhaustion. Third, throughout the cross-level analysis, the result indicated that PsyCap had no moderating effect on the relationship between workplace incivility and emotional burnout. PsyCap, on the other hand, was found to have significant negative effects on emotional exhaustion, and so did perceived service climate. Fourth, service climate had negative impacts on emotional burnout and, finally, service climate was found significantly positively related to PsyCap.According to the findings, suggestions have been made for managerial implications. As for frontline employees at sport and fitness centers, it is suggested to use the facilities at the workplace which can improve their health and decrease emotional exhaustion. As for workplace incivility, the human resources department may establish appropriate procedures to assist employees in coping with difficult people to decrease employees’ emotional exhaustion. As for PsyCap, people were not born with PsyCap. In fact, PsyCap can be developed through learning. Therefore, for any individual or organization, the recognition and development of PsyCap could help develop better self-awareness . IndividUpon the first day of the participants’ recruitment, all respondents promised to complete the daily investigation. However, there may have been some tired-out employees forgetting to complete the daily investigation and leaving their responses to the very next day, or even just simply using the next day’s perception. In any case, we strived to encourage the participants to complete the daily investigation to avoid this confusion.


Levels of O2•− were significantly higher in radiation exposed workers compared to control. Similarly, a significant increase in the levels of interleukin (IL)-6, IL-1α and macrophage inflammatory protein (MIP)-1α in radiation exposed workers compared to control was observed, while there was no significance difference in the other 27 screened cytokines. A significant positive correlation was found between MIP-1α and O2•− levels with no correlation in either IL-6 or IL-1α. Further, a dose-dependent relationship with significant O2•− production and immune alterations in radiation exposed workers was demonstrated. There was no statistical difference between the groups in terms of oxidative DNA damage, GSH/GSSG levels, or EcSOD activity. Although the biologic significance of cytokines alterations in radiation exposed workers is unclear, further studies are needed for determining the underlying mechanism of their elevation.Studies have shown an increased risk for a variety of cancers, specifically brain cancer, in healthcare workers occupationally exposed to ionizing radiation. Although the mechanisms mediating these phenomena are not fully understood, ionizing radiation-mediated elevated levels of reactive oxygen species (ROS), oxidative DNA damage, and immune modulation are likely involved. A group of 20 radiation exposed workers and 40 sex- and age-matched non-exposed control subjects were recruited for the study. We measured superoxide (O Ionizing radiation is used commonly in medical diagnostics, and the advancement of diagnostic imaging and interventional radiology has raised concern about the potential risk these advancements may pose to healthcare workers utilizing these technologies. Today, surveillance of healthcare workers chronically exposed to ionizing radiation only provides information on accidental overexposure, not on the real chronic risk of exposure to low dose ionizing radiation. Therefore, there is an urgent need to closely examine potential pathological changes occurring in workers chronically exposed to ionizing radiation. Although occupational exposure to ionizing radiation generally falls well below the currently accepted limits ) set by the International Commission of Radiation Protection (ICRP) , several2•−) [We previously reported that occupational exposure to ionizing radiation even within the limits of ICRP recommendations results in an alteration of redox environment with an increase in ROS, particularly superoxide (O2•−) . Further2•−) , as well2•−) ,10,11. I2•−) ,4 and di2•−) . Strong 2•−) ,4,12. It2•− levels in radiation exposed workers. In view of the above considerations, the present study was designed to assess the plasma inflammatory cytokines level, oxidative DNA damage, and antioxidants to determine their association with O2•− levels in radiation exposed workers at different occupational settings.A growing body of evidence indicates immunological changes induced by exposure to ionizing radiation. The effect of ionizing radiation on selected indices of cellular and humoral immunities in workers occupationally exposed to low levels of ionizing radiation has been studied ,14. Clusn = 12), Interventional Radiography , and Computed Tomography , while the remaining 40 samples were from unexposed individuals’ age- and gender matched  tubes. Blood plasma and red blood cells were separated by centrifugation of the whole blood at 2500× 2•− levels were assessed as described in our earlier study [Total cellular Oer study . 100 μL TM Oxidative DNA Damage ELISA Kit , and per the manufacturer’s instructions. Concentrations were expressed as ng/mL. The assay detection sensitivity ranges from 100 pg/mL to 20 ng/mL.DNA oxidation (8-hydroxydeoxyguanosine (8-OHdG)) was assessed in blood plasma using commercially available assay, OxiSelect® Human Cytokine 30-Plex Kit  per the manufacturer’s instructions. Plates were then analyzed by the Meso Scale Discovery® (MSD) QuickPlex SQ 120 and samples concentrations were calculated using the Discovery Workbench 4.0 using a 4-PL curve fit model. Samples below the lower level of detection, which was calculated by the workbench software as 2.5 standard deviations above the assay background blank, were reported as 0 pg/mL.Electrochemiluminescence-based immunoassay was used to measure cytokine levels in plasma. Samples and standards were prepared on multispot 96-well plates from the V-PLEXReduced glutathione (GSH) and oxidized glutathione (GSSG) Levels in blood plasma were measured using a commercially available assay . Activity of EcSOD in plasma was measured using a superoxide dismutase (SOD) Assay Kit from Dojindo , according to the manufacturers’ guidelines.p value < 0.05 was considered significant for all statistical analyses.GraphPad Prism version 5  was used for statistical analysis. Data are presented as mean ± standard error of the mean (SEM). Comparison between two groups was performed with the Mann- Whitney test. Comparison between three or more groups was done by One-way analysis of variance with Bonferroni post hoc tests. Pearson’s correlation test was performed to identify the relationships between variables. A Demographic characteristics of the study subjects are not statistically different between control and radiation exposed workers as shown in 2•− level in whole blood of radiation exposed workers was significantly higher compared with the control subjects, p < 0.05, p > 0.05, As we previously reported , O2•− lep < 0.05) different betweeen radiation exposed workers and control subjects , whereas both IL-6 and IL-1α did not   .p > 0.05). Moreover, there were insignificant differences in plasma 8-OHdG between CR, IR and CT subgroups, (p > 0.05). There was no correlation between 8-OHdG and O2•− levels.The results of plasma 8-OHdG in radiation workers subgroups compared to control group are illustrated in 2•− in radiation exposed workers, it would be expected to observe a difference in antioxidant levels in radiation workers compared to control group. We have previously [p > 0.05, 2•−. Similarly, no significant difference was observed between the two study groups in GSH/GSSG ratio . However, the association of GSH/GSSG with O2•− level failed to demonstrate any association .Owing to elevated levels of Oeviously  reported2•− levels using EPR spectroscopy. Our data reveal that in IR and CT subgroups (high dose radiologic procedures) compared to CR subgroup (low dose radiologic procedures), chronic exposure to ionizing radiation is associated with an increase in O2•−. Considering we used whole blood for these measurements, it is likely that lymphocytes and/or erythrocytes are the source of increased levels of O2•−.The effect of occupational exposure to low levels of ionizing radiation is a serious concern to a large number of radiation workers . Althoug2•− levels with no correlation in either IL-6 or IL-1α. Our current understanding of the importance of immune system’s role in cancer control, studies of radiation-immune system interactions have been one of the main research fields in radiation biology and radiation protection. However, there has been little research examining the dose–response relationship of ionizing radiation-induced immune alterations. Interestingly, our data has showed a significant increase in MIP-1α in the plasma of radiation exposed workers especially in IR subgroup (high-dose radiologic procedures), compared to CR subgroup (low dose radiologic procedures) and control, which suggests an association between inflammation and high radiation absorbed dose in the radiation exposed workers. Acute inflammation is the initial protective response by the body; however, chronic inflammation can lead to pathology. Proper regulation of cytokine production is critical in diseases control and prevention [2•− levels [Our body has a variety of defense mechanisms comprising antioxidant enzymes to counteract ROS-mediated oxidative damage ,26. Therevention . In our evention  as discu− levels  and shou− levels . Hrycek − levels . In addi− levels . Another− levels . Shieh e− levels . AlthougWith respect to DNA oxidation, the current study showed that the mean values of plasma 8-OHdG in all radiation exposed workers compared to control group were insignificant, with no significant values observed among IR compared to CR or CT . This fiIn general, the link between radiation-induced DNA damage and immune response has been demonstrated in many cases as reviewed in ,36. Prev2•− and lipid peroxidation [2•− into H2O2 to reduce ROS-mediated diseases such as carcinoma, inflammation, and aging [Earlier, we demonstrated that occupational radiation exposure within ICRP recommendation limits , resultsxidation . These fxidation . The curxidation . GSH is xidation ,41,42,43xidation . Therefoxidation ,46, it sxidation . Superoxnd aging ,49. Prevnd aging ,46. Our 2•−, as there is evidence that it reacts with peroxynitrite, nitrogen dioxide, and peroxyl radical. However, it should be noted that O2•− has the highest interaction constant with CMH [2•− with CMH and this supports our conclusion that there is increased oxidative stress in our studied subjects. Lastly, we posit that the increase in O2•− levels observed in whole blood is likely from lymphocytes; however, without knowing the exact white blood cells profile in our samples, it is difficult to make strong conclusions regarding the precise source of O2•−.It must be noted there are limitations in our study. A larger sample size would allow more statistical significance in the evaluation of biomarkers. Also, using 8-OHdG to estimate DNA damage in humans after ionizing radiation exposure is another limitation in our study as plasma 8-OHdG levels could be influenced by the rate of repair as well as by the rate of damage and ultimately we need to use other techniques in the future to assess DNA damage at nuclear and cellular levels. Further, CMH is not 100% specific for Owith CMH ,51. As s2•− production and immune alterations in radiation exposed workers and more specifically in high dose radiologic procedures . Although the biologic significance of changes in these cytokines is unclear, results from the current study indicate the importance to take all necessary measures to protect radiation exposed workers from radiation exposure. The underlying mechanism of their elevation needs further investigation. In view of the importance to improve understanding of the long-term health effects in workers occupationally exposed to radiation, Low dose radiation effect studies have to be one of the main research priority. Thus, follow-up evaluation of occupational health status, should be considered an integral part of quality assurance programs.In conclusion, the results presented in this study have demonstrated a dose-dependent relationship with significant O


Taken together, this joint review of public literature and omics data records points to two members of the acyl-CoA synthetase family potentially playing a role in inflammasome activation in neutrophils. Translational relevance of these observations in the context of sepsis and other inflammatory conditions remain to be investigated.A potential role for the long-chain acyl-CoA synthetase family member 1 (ACSL1) in the immunobiology of sepsis was explored during a hands-on training workshop. Participants first assessed the robustness of the potential gap in biomedical knowledge identified via an initial screen of public transcriptome data and of the literature associated with ACSL1. Increase in ACSL1 transcript abundance during sepsis was confirmed in several independent datasets. Querying the ACSL1 literature also confirmed the absence of reports associating ACSL1 with sepsis. Inferences drawn from both the literature (via indirect associations) and public transcriptome data (via correlation) point to the likely participation of ACSL1 and ACSL4, another family member, in inflammasome activation in neutrophils during sepsis. Furthermore, available clinical data indicate that levels of ACSL1 and ACSL4 induction was significantly higher in fatal cases of sepsis. This denotes potential translational relevance and is consistent with involvement in pathways driving potentially deleterious systemic inflammation. Finally, while ACSL1 expression was induced in blood  Long-chain acyl-CoA synthetases (ACSLs) are essential enzymes that activate fatty acids (FA) by converting them to FA acyl-CoA esters. This activation is required for both synthesis of cellular lipids, such as triacylglycerol (TAG), phospholipids, and cholesterol esters as part of anabolic lipid metabolism, as well as their degradation via β-oxidation as part of catabolic lipid metabolism . Deregulin vitro exposure to septic plasma, and on the other of the absence or limited number of literature reports linking it with neutrophil immunobiology or the pathogenesis of sepsis.Involvement of ACSL1 in immunological processes was examined in the context of a hands-on workshop which used available public omics data as source of training material. ACSL1 was selected among a pool of gene candidates on the basis, on one hand of its upregulation in neutrophils following Focusing a review on a research area which has been overlooked is inherently challenging given the lack of good published reports on the topic in question. However, exploring such gaps in knowledge and attempting to define avenues for future research remains a useful endeavor. Furthermore, publicly available omics data were employed as a means to, at least in part, overcome the absence of publication on the topic being covered. Indeed, in addition to initially permitting identification of the knowledge gap this approach also provided opportunities: (1) to confirm initial observation across independent datasets, (2) to draw inferences from co-expression (guilt by association). While this review does not address the identified knowledge gap in a way a research paper would, it permits to bring such a gap to the attention of the community in a timely manner and delineates potential avenues for future investigations.As described in a recent review, workshop participants for the first of three “collective omics data” (COD) training modules currently under development are tasked to assess potential gaps in biomedical knowledge for a given gene candidate .in vitro to plasma of septic patients  = text words: thus restricting the search to text included in tiles and abstracts]:ACSL1” [tw] OR “Acyl-CoA Synthetase Long Chain Family Member 1” [tw] OR “Acyl-CoA Synthetase Long Chain Family Member 1” [tw] OR “Fatty-Acid-Coenzyme A Ligase, Long-Chain 2” [tw] OR “Long-Chain Fatty-Acid-Coenzyme A Ligase 1” [tw] OR “Long-Chain-Fatty-Acid–CoA Ligase 1” [tw] OR “Long-Chain Fatty Acid-CoA Ligase 2” [tw] OR “Long-Chain Acyl-CoA Synthetase” [tw] OR “Long-Chain Acyl-CoA Synthetase 1” [tw] OR “Long-Chain Acyl-CoA Synthetase 2” [tw] OR “Lignoceroyl-CoA Synthase” [tw] OR “Palmitoyl-CoA Ligase 1” [tw] OR “Palmitoyl-CoA Ligase 2” [tw] OR “Acyl-CoA Synthetase 1” [tw] OR “LACS 1” [tw] OR “LACS 2” [tw] OR “LACS-1” [tw] OR “LACS-2” [tw] OR “FACL2” [tw] OR “FACL1” [tw] OR “LACS1” [tw] OR “LACS2” [tw] OR “ACS1” [tw] OR “LACS” [tw] OR “Fatty-Acid-Coenzyme A Ligase, Long-Chain 1” [tw] OR “Palmitoyl-CoA Ligase 1” [tw] AND “This PubMed query returned a total of 625 articles as of May of 2019. The query argument, following the boolean “AND” was introduced to minimize the rate of false positives. It excluded irrelevant articles including for instance the ACSL1 alias “LACS” but referring to something entirely different, such as “Laser-Assisted Cataract Surgery” (LACS), or “Low molecular weight Aromatic Compounds” (LACs).Next, the degree of novelty of the observation reported in in vitro to septic plasma, resulting in significant increase in levels of abundance of ACSL1 transcripts. Therefore, the most relevant topics selected for the assessment of potential gaps in biomedical knowledge were “sepsis” and “neutrophils.”The study that served as a starting point for this review consisted in exposing neutrophils One of the 625 articles constituting the ACSL1 literature contained the keywords “sepsis,” “septic,” “septicemia,” or “septiceamia,” This work examined responses to endotoxin treatment in mice and made reference to sepsis for context without actually studying septic animals . The autIt was thus possible to conclude at this stage:in vitro to septic serum is a relevant phenomenon in vivo, since it was also increased in the blood of septic patients.- First that the increase in ACSL1 abundance observed in neutrophils exposed - Second, that this phenomenon can be considered to be robust since it was observed in multiple studies across various clinical and study settings.- Third, that an increase in ACSL1 expression has not been reported in the literature in the context of sepsis to date. And that a role for ACSL1 in neutrophil immunobiology has been described, but not in the context of sepsis. Furthermore, it is an area of research which has been inactive for at least the past two decades.The next steps consisted in bringing additional context via a general review of the literature retrieved for ACSL1 and four other members of the long chain acyl-CoA synthetase family. Several reference transcriptome datasets which are available publicly were also consulted in order to complete this picture.In order to draw inferences on the possible role played by ACSL1 in sepsis, context was obtained by conducting a background literature review. As a first step, each of the workshop participants were tasked to retrieve literature for one of the five known members of the family of human ACSL genes. Once they had optimized PubMed queries designed to retrieve the literature of each respective family members, keywords identified “manually” in titles of articles were recorded and categorized . Recording the frequency at which those keywords occurred in the literature allowed in turn the identification of the main research themes for each family member.Furthermore, tissue-specific expression of ACSL family members was looked up in BioGPS, a publicly accessible data browsing portal which comprises several reference transcriptome datasets . Relativlipid metabolism, fatty acid uptake, insulin, and muscle. ACSL1 is the main ACSL family member expressed in the liver (CCL2) and chemokine (C-C motif) ligand 5 (CCL5) and the macrophage associated surface antigen cluster of differentiation 14 (CD14)  . Althoug4 (CD14) . A simil4 (CD14) . Upregul4 (CD14) , colon c4 (CD14) , and liv4 (CD14) , 33, 34.4 (CD14) , 26 (Sup4 (CD14) . Intereslipid droplet, liver, and goose fatty liver. Expression of ACSL3 has been described in various tissues including the brain   . ACSL exN = 19) . VariousN = 19) –99.Interestingly, despite the limited number of publications linking the activity of the long chain acyl-CoA synthetases to immunity, members of the family were found to be robustly expressed across various immune cell subsets . While ALegionella pneumophila, a flagellated gram negative bacteria, was the most potent inducer of ACSL4 expression in this culture system. PAM3 [Pam (3) CSK (4)] is a strong stimulator of TLR5 expression in neutrophils . A high degree of correlation was also observed among the transcripts forming M13.12 in the septic plasma neutrophil exposure dataset used as a starting point for this work . Here we took advantage of a repertoire of transcriptional modules developed by our team on the basis of co-expression analysis of whole blood transcriptome profiles obtained for 16 disease states across nearly 1000 subjects. Such repertoires of co-expressed gene sets have been in use for over 10 years , 117, anhis work . CorrelaCorrelations were also measured for two “canonical” inflammasome and fatty acid metabolism gene set (identified via a knowledge-driven approach for the purpose of offering targeted “RT2 profiler PCR arrays”). As shown on Taken together, inferences that could be drawn based on both profiling of the literature and transcript co-expression analysis confirm a likely role of ACSL1 in driving inflammasome-mediated release of pro-inflammatory factors by neutrophils during sepsis.The burden of sepsis remains high, with poor outcomes observed especially in developing countries. Despite several attempts at developing drugs that would control overt inflammation no immune-modifying treatment is currently available for the management of sepsis . Contribin vitro in response to septic serum . The panel of stimuli employed in the experiment described in  context , 27] in https://casrai.org/credit/).JR, MG, DR, AJ, DB, and DC: conceptualization. MG, MT, and DR: data curation and validation. JR, DR, and DC: visualization. JR, MG, EH, YM, PS, and DC: analysis and interpretation. JR and DC: writing of the first draft. SB, DB, and DC: funding acquisition. MG, MT, DR, and DC: methodology development. MA, MT, and SB: software development and database maintenance. JR, MG, EH, YM, PS, MT, MA, WH, SB, DR, AJ, DB, and DC: writing–review and editing. The contributor's roles listed above follow the Contributor Roles Taxonomy (CRediT) managed by The Consortia Advancing Standards in Research Administration Information (CASRAI) (The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


N = 270), including 44 participants with MH, asking participants to rate imagery, earworms, or MH on several variables. MH were reported as occurring less frequently, with less controllability, less lyrical content, and lower familiarity, than other forms of inner music. MH were also less likely to be reported by participants with higher levels of musical expertise. The findings are outlined in relation to other forms of hallucinatory experience and inner music, and their implications for psychological models of hallucinations discussed.Musical hallucinations (MH) account for a significant proportion of auditory hallucinations, but there is a relative lack of research into their phenomenology. In contrast, much research has focused on other forms of internally generated musical experience, such as earworms (involuntary and repetitive inner music), showing that they can vary in perceived control, repetitiveness, and in their effect on mood. We conducted a large online survey ( Although the most frequently reported form of AH are auditory verbal hallucinations (AVH), phenomenological surveys have also shown that a substantial minority of people also report musical hallucinations (MH): that is, the perception of music when none is playing. For example, one survey of 100 people with psychosis and AVH found that 36% also described the occurrence of MH . The mosSurveys focusing exclusively on MH have suggested that they may occur in around 16% of individuals with a diagnosis of schizophrenia , and as Furthermore, it is unclear to what extent the attributes assessed in the small amount of previous research could also be applied to other forms of ‘inner music’To our knowledge, no research has directly compared self-reported experiences of musical imagery and earworms to MH, and, in fact, the boundary between earworms and MH is somewhat unclear in much of the literature. For example, Hemming cited in  defines Research into musical imagery and earworms has also investigated their effect on mood and behavior, but again, these have not been directly investigated in comparison to MH. For example, There are, then, several large gaps in the MH literature, which we sought to address in the present study. Firstly, very little research has been conducted regarding the phenomenology of MH much further than asking about the broad style of music experienced. Secondly, there is some confusion within the literature over precise definitions as to what constitutes an MH, leading previous studies to use different inclusion criteria. As mentioned, https://doi.org/10.1016/j.concog.2018.07.009.Supplementary data associated with this article can be found, in the online version, at We sought to conduct an exploratory survey of the phenomenology of MH, to investigate potential similarities and differences between MH and other forms of inner music. Participants were asked to pick a category that they felt best described their inner musical experience  based on basic definitions see , or spec22.1http://hearingthevoice.org/2014/11/04/round-and-round-the-phenomenology-of-inner-music). Rather than aiming to recruit a sample representative of the national population, the aim was to recruit participants who reported hallucinatory experiences or regular inner music, to investigate phenomenological features of these experiences.Participants were invited to take part in an on-line survey, advertised via social media and a research project website . The majority of participants were from English-speaking countries  but there were also respondents from other European countries  and non-European countries around the world . Some form of hearing loss was reported by 15.8% of participants, with most of these being described simply as hearing loss/hypacusis (80%) and/or tinnitus (37.5%). Mean reported length of hearing impairment in these individuals was 18.3 years (SD = 17.3). 42.7% of participants reported having previously received a psychiatric diagnosis, with the largest proportion of these being for depression (60.6%) or anxiety (30.3%), with smaller numbers of participants having diagnoses of bipolar disorder, obsessive compulsive disorder, attention deficit hyperactivity disorder, or schizophrenia/schizoaffective disorder/psychosis. 9.0% of participants reported some form of neurological disorder, whilst 26.4% reported being on some form of medication for a psychiatric/neurological disorder. See There were 276 respondents to the questionnaire. From these, 7 participants were excluded who did not respond to a sufficient number of questions (<10% response rate) on the ‘Phenomenology of Inner Music Questionnaire’ (see below), whilst 14 that described an alternative form of inner music in the free text box  were excluded. Thus, the sample analyzed consisted of 255 participants , with a mean age of 39.4 , familiarity, effects on mood and behavior, amount of perceived control, triggers, and likelihood of being mistaken for an external stimulus. All questions required the participant to respond on a Likert scale, although many questions also provided a free text box to elicit more detailed responses. (See 2.2.3I have thoughts that I cannot stop) and thought suppression . For each question, participants are required to rate their agreement on a Likert scale from 1 (Strongly Disagree) to 5 (Strongly Agree). These subscales have previously been shown to have acceptable internal reliability . For each question, participants are required to indicate agreement with each question on a 4-point Likert scale, ranging from 1 (Never) to 4 (Almost Always). Total score can range from 5 to 20. It has previously shown high internal reliability (Cronbach’s α = .73)  .2.2.5I think in inner speech about what I have done, and whether it was right or not), dialogic inner speech , other people in inner speech  and condensed inner speech . Each item is scored on a Likert scale from 1 (Never) to 6 (All of the time). Each subscale has previously shown high internal reliability (Cronbach’s α > .8) and acceptable test-retest reliability (>.6)  .2.3k = .77), and any disagreements (n = 9) were resolved by discussion between authors. Thus, the sample was split into four groups  for between-group analysis.Given that our main area of interest regarded the phenomenological differences between MH and other forms of inner music , participants were categorized by the main type of experience they reported. If participants indicated in the free text box that they frequently experienced more than one form of inner music , but indicated in the free text box that one of these was much more prevalent than the other, they were categorized according to their most prevalent experience. If participants indicated that they experienced more than one type of music, but did not report relative frequencies, they were categorized in a separate ‘mixed experiences’ group. This categorization was performed separately by two of the authors  to limit the number of tests performed. Based on our areas of interest, we split the analysis into four main sections: (1) demographic and etiological information; (2) basic characteristics and acoustic details of inner music; (3) location and controllability of inner music; (4) effect of inner music on behavior and mood. Qualitative examples given by participants are included throughout as illustrative examples of different aspects of their phenomenology.  Bonferroni corrections for multiple comparisons were applied within each section  in conjunction with 95% confidence intervals. Due to mainly ordinal and non-normally distributed data, non-parametric ANOVAs  were used to test for differences between the categories of inner music, for each phenomenological attribute. Where appropriate, Mann-Whitney 33.1The first question of the Inner Music Questionnaire asked people to choose the category that best described the music they heard. Of the 255 participants, 17.3% reported musical hallucinations, 40.4% reported earworms, 27.1% reported musical imagery, whilst 15.3% were categorized in the ‘mixed experiences’ group. To investigate phenomenological differences between MH and other musical experiences, ‘inner music category’ was used as a between-subject variable. Only 1.2% of participants reported that they could ‘Never’ accurately describe their inner music (in response to Q5) but, since they continued to provide responses to the questions, these participants were not excluded from the sample. 2(4) = 4.33, p = .228, φ = .131) or neurological diagnosis (χ2(3) = 1.36, p = .714, φ = .073) and the category of inner music. There was also no significant association between presence of a hearing impairment and category of inner music (χ2(3) = 0.89, p = .828, φ = .059).A chi-square analysis indicated that there was not a significant association between presence of a psychiatric diagnosis (χ2(15) = 36.93, p = .001, φ = .381), although in this analysis 25% of cells had an expected count of < 5, due to the low number of semi-professional (n = 23) or professional (n = 9) musicians (at least compared to other groups) in the sample. This violates a key assumption of chi-square analysis (n = 110) and musicians . Again, a chi-square analysis indicated an association between musical expertise and inner music type (χ2(3) = 20.99, p < .001, φ = .287). Further analysis suggested that musicians were less likely to report MH , but more likely to report mixed experiences  compared to non-musicians. There was little difference in the proportion of musicians in either the imagery  or earworm  groups. Non-parametric ANOVAs  with inner music category as the independent variable suggested a similar pattern of results for number of hours spent practicing musical instruments per week (χ2(3) = 17.82, p < .001), with MH being associated with less music practice than imagery , although not significantly different to earworms  or mixed experiences  at the corrected alpha level. However, this pattern of results seemed to be specific to actually practising music, and did not hold for time spent listening to music (χ2(3) = 2.23, p = .526).There was a significant association between level of musical expertise and type of inner music reported , frequency (Q2), duration (Q3), familiarity (Q10), repetitiveness (Q22), and whether music was experienced in its entirety or was shortened (Q20 and 21). Participants were also asked whether their inner music included attributes such as melody, harmony, intensity, instruments, and lyrics (Q6). Classical music was the most frequently reported style of inner music in participants with MH (47.7%), whereas rock music was the most frequently reported style in the musical imagery group (64.2%), and pop music in the earworms group (66.0%). The mixed experiences group reported that both pop music and classical music were the most frequent style of their inner music (both 62.5%). See 2(3) = 33.73, p < .001), with Mann-Whitney U tests  indicating that participants in the MH group reported the experience as occurring significantly less frequently than those in the imagery group , earworm group , or the mixed experiences group . There was also a significant main effect of inner music type on familiarity (χ2(3) = 56.96, p < .001), with MH being rated as less familiar than imagery , earworms , or mixed experiences . For example, a typical response from a participant in the MH group in a free-text box was:A non-parametric  ANOVA with inner music frequency  = 21.22, p < .001), with MH being reported as less repetitive than earworms  or imagery , but not significantly different to the mixed experiences group , following corrections to the alpha level (.05/3 = .017). There was also no significant effect of inner music type on duration (χ2(3) = 11.60, p = .009), or the extent to which participants reported inner music being shortened (χ2(3) = 5.31, p = .151) or a complete song (χ2(3) = 5.77, p = .123).There was also a main effect on repetitiveness (χ2(3) = 3.02, p = .389, φ = .109), intensity (χ2(3) = 5.12, p = .163, φ = .142) or instruments (χ2(3) = 5.69, p = .128, φ = .149) were perceived, although there was an association between inner music type and whether lyrics were perceived (χ2(3) = 23.02, p = < .001, φ = .300).  Further analysis showed that MH were less likely to include lyrics than other types of inner music , whereas earworms were more likely to include lyrics . Meanwhile, the imagery  and mixed experiences , with most also being able to perceive harmony (77.4%), intensity (66.5%), instruments (77.0%) and lyrics (75.9%). Chi-square analysis indicated no association between inner music category and whether harmony (χ3.3Further questions asked whether participants felt control over their inner music (Q23), whether they could anticipate the experience (Q24), whether they knew the trigger of the experience (Q25), whether they ever mistook it for an externally located stimuli (Q26), and whether they felt like the experience was of their own creation (Q11). 2(3) = 11.52, p = .009), with participants reporting MH also reporting less control over the experience compared to participants in the imagery group . However, there was no significant difference between perceived control of MH and earworms, or between MH and mixed experiences (ps > .129). A typical comment related to the perceived effortlessness of MH, with little or no control:Imagination to me is a conscious effort – this isn’t.”“Like a radio station, I just have to wait for the next song.”“There was a main effect of category of inner music on perceived control .”“Others, meanwhile, described techniques to stop the MH, such as distracting themselves with another activity:2(3) = 95.10, p < .001), with MH being more likely to be mistaken for an external percept than imagery , earworms  and mixed experiences . For example:“There is usually a moment where I am not entirely sure if it is external or internal, but the quality of sound and the apparent feeling of “proximity” is strange when it is a hallucination. In other words, it might sound soft as if it should be coming from far away, and yet it does not sound as though it is coming through any barriers like walls... it is almost as if it is coming through earphones, closer to me than the outside world, but not exactly ‘in my head’”Remembering music is like a faint shadow of real music...whereas hallucinating it really involves hearing it”“When asked about the frequency with which their inner music was mistaken for coming from the external environment, there was a significant main effect of inner music category (χ2(3) = 39.09, p < .001), with participants in the MH group, counterintuitively, rating this attribute more highly than participants in the earworm group , although there was no difference between the MH and imagery or mixed experiences groups at the corrected alpha level (ps > .30).There was a significant main effect of inner music type on the extent to which participants reported that their inner music was their own creation , whether their body moved with the music they experienced (Q9), whether the experience reflected their own feelings (Q18), or was associated with negative affect (Q15) see . A Bonfe2(3) = 19.22, p < .001), with MH being rated lower on this attribute than imagery , earworms  or the mixed experiences group . Although there was a significant main effect of inner music category on the extent to which the experience made the participant feel excited (χ2(3) = 15.30, p = .002), further tests did not reveal any significant differences at the corrected alpha level between MH and any other categories . Notably, there was no significant effect of inner music category on the experience contributing to the participant feeling depressed (χ2(3) = 8.21, p = .042) or anxious (χ2(3) = 6.79, p = .079) after corrections to the alpha level, providing no strong evidence that MH were associated with negative affect more than other forms of inner music. For example:It’s never anything emotional playing, usually just ‘there’. I’ve gotten used to it for the most part, but the most it makes me feel is annoyed.”“The extent to which participants reported that the inner music reflected how they were feeling differed between categories (χ2(3) = 24.72, p < .001), compared to imagery , earworms , or mixed experiences , as well as being less able to hum the music after the experience (χ2(3) = 22.15, p < .001) compared to imagery , earworms , or mixed experiences . Participants also reported moving their body less to MH (χ2(3) = 28.88, p < .001), compared to imagery , earworms , or mixed experiences . One typical comment was:Unfamiliar, no lyrics, no compulsion to sing or hum along”“Participants reported being less able to hum along with MH . Unsurprisingly, there was a main effect of inner music type on hallucination-proneness (χ2(3) = 20.03, p < .001), with participants in the MH category  scoring higher than those in the imagery category  and the earworm category , although not significantly differently to the mixed experiences category .Participants also completed a small number of quantitative self-report measures relating to phenomenology of inner speech (VISQ), intrusive thoughts and thought suppression (WBSI), and auditory hallucination-proneness (LSHS). There was no main effect of inner music type on any measure of inner speech phenomenology, intrusive thoughts, or thought suppression (all 4The present study provided preliminary evidence for a number of phenomenological differences between MH and other forms of inner music (summarized in A previous systematic review by Indeed, the only demographic factor significantly associated with MH in the present data was level of musical expertise, with individuals reporting MH being less likely to classify themselves as a musician, playing fewer musical instruments, and spending less time practising music. Previous research has suggested that musical training is associated with superior performance on tasks requiring auditory imagery generation, for example to complete a musical sequence , or to eOne of the few studies to report phenomenological details of MH  reports Another clear difference between earworms and MH, in our sample, was the reported familiarity of the perceived music, with MH being rated as less familiar to the participant than other forms of inner music. It is possible that this feeling of unfamiliarity may add to a feeling of alienness typically associated with hallucinations, as opposed to musical imagery or earworms. In contrast, previous studies have reported that as many as 78% of cases of MH were experienced as familiar music . There aOur data suggested that less than half of participants reported the presence of lyrics in their MH, compared to rates of approximately 80% in other forms of inner music. This is, again, in contrast to Saba and Keshavan, who noted that MH consisting only of instrumental music was rare. It should be noted that the present questionnaire only asked about lyrical content rather than concurrent MH and AVH. Nevertheless, the low frequency of lyrical content was an unexpected finding, and highlights a key difference between MH and AVH, the most frequent form of auditory hallucination. Given that cognitive neuroscientific models of AVH specify a key role for brain networks involved in speech production and perception , this isA further unexpected, and rather counterintuitive, aspect in which MH differed from other forms of inner music was in rating the extent to which the music was one’s own creation. Although MH were rated as lower in controllability and familiarity, they were actually rated as significantly higher on this attribute, in comparison to earworms. A frequent argument is that earworms occur due to unintentional re-activation of memory representations of previously heard music . In thisThis study, then, has provided data on several aspects of the phenomenology of MH. In comparison to the most widely studied type of inner music – earworms – MH are less controllable, more likely to be experienced as coming from the external environment, less familiar in content, less repetitive, less likely to include lyrics, and more recognizable as one’s own creation. These differences highlight a need for greater clarity in the use of definitions when talking about different experiences of inner music. Our data is not consistent with previous claims that MH can simply be thought of as earworms that have reached pathological levels (e.g., Hemming, cited in Future research should further investigate some of the findings presented here. One limitation of the present study was that it required participants to classify their own experience as hallucinatory. Ideally, such experiences should be enquired about via a face-to-face clinical interview; however, online surveys can reveal experientially rich and sometimes unexpected aspects of hallucinatory phenomenology (see, for example, None.


The article “The metabolic cost of nesting: body condition and blood parameters of Caiman crocodilus and Melanosuchus niger in Central Amazonia”, written by José António Lemos Barao‑Nobrega, Boris Marioni, Robinson Botero‑Arias, António José Arsénia Nogueira, Emerson Silva Lima, William Ernest Magnusson, Ronis Da Silveira, Jaydione Luiz Marcon was originally published electronically on the publisher’s internet portal (currently SpringerLink) on June 19, 2017 without open access.With the author(s)’ decision to opt for Open Choice the copyright of the article changed on November 1, 2017_The Author(s) [2017] and the article is forthwith distributed under the terms of the Creative Commons Attribution [continuing for CC BY license].http://creativecommons.org/licenses/by/4.0/), which permits use, duplication, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.4.0 International License (The original article has been corrected.


Major depressive disorder (MDD) is a complex, heritable psychiatric disorder. Advanced statistical genetics for genome-wide association studies (GWASs) have suggested that the heritability of MDD is largely explained by common single nucleotide polymorphisms (SNPs). However, until recently, there has been little success in identifying MDD-associated SNPs. Here, based on an empirical Bayes estimation of a semi-parametric hierarchical mixture model using summary statistics from GWASs, we show that MDD has a distinctive polygenic architecture consisting of a relatively small number of risk variants (~17%), e.g., compared to schizophrenia (~42%). In addition, these risk variants were estimated to have very small effects (genotypic odds ratio ≤ 1.04 under the additive model). Based on the estimated architecture, the required sample size for detecting significant SNPs in a future GWAS was predicted to be exceptionally large. It is noteworthy that the number of genome-wide significant MDD-associated SNPs would rapidly increase when collecting 50,000 or more MDD-cases (and the same number of controls); it can reach as much as 100 SNPs out of nearly independent (linkage disequilibrium pruned) 100,000 SNPs for ~120,000 MDD-cases. Major depressive disorder (MDD) is a common, complex disorder with a high lifetime prevalence of ~15%  and almost all SNPs were within |β| = 0.04 (odds ratio = 1.04). For ASDs, effect sizes were estimated to be relatively small among the six GWASs; almost all SNPs were within |β| = 0.05. For CD, we had many disease-associated SNPs with effect sizes near or more than |β| = 0.05 or odds ratio = 1.05, and also peaks of effects around |β| = 0.1. The estimated distribution of g for SCZ lay mostly within a range of |β| ≤ 0.03, but with peaks at relatively large effects of |β| 0.05 or larger. AN had relatively large effects, particularly in the positive signed region. For T2D, while most disease-associated SNPs were within |β| = 0.03, there was a small portion of disease-associated SNPs with the effect sizes near or more than |β| = 0.05.Non-parametric estimation of pc = 5 × 10−8 . It can reach as much as 100 SNPs out of 100,000 independent SNPs for ~120,000 MDD-cases Figure . Note thP-value) are preferentially selected among “all SNPs” so that linkage disequilibrium (LD) among selected SNPs are nearly independent). The discrepancy between our prediction and the observation could be due to the difference between the PGC cohort data and data from the other six cohorts, especially, self-reported data from 23andMe with 130,664 cases and 330,470 controls, which accounted for the large proportion of the total cohorts. In fact, the SNP heritability estimates in observed scales were much smaller for 23andMe data (0.038) than for PGC data (0.128)  including PGC data with 16,823 cases and 25,632 controls β2for Park et al. ” in the discretized effect size distribution used in the estimation algorithm). We have shown that with 3–5,000 or more cases (and the same number of controls), the estimates of π and g are fairly accurate, leading to reliable estimates of the number of significant disease-associated SNPs  will be addressed in future work. Although we assumed 100,000 SNPs in the LD-pruned set from the observations in Table Me; the key determinant of the accuracy of genomic prediction) does, i.e., Me = 2.938 Ne is the effective population size . In modeling the non-null component (effect size distribution), the parametric approach, e.g., finite normal mixture models with several components, is a popular choice. Unlike such a parametric model, we assume a non-parametric distribution as a “single” non-null component to cover all such non-null components. This is the interpretation for the modeling formula given in the subsection “Semi-parametric Hierarchical Mixture Model (SP-HMM)” in the Materials and Methods section. Meanwhile, in estimation using the expectation–maximization (EM) algorithm we can see our model as that with “so many” non-null components  needed to be collected in a single cohort study, or additional cohorts (sample sets) needed to be included in a meta-analysis, and for discovering a given number of new disease-associated variants.g, non-parametrically, using GWAS summary statistics on effect sizes (genotype log-odds ratios) which often are available through Web sites. The “non-parametric estimation of g” enables us to flexibly characterize the effect-size distributions without any assumptions for forms of the distribution. The SP-HMM assumes independence among SNPs, as was justified by pruning SNP described below. The SP-HMM has been validated through various types of polygenic scenarios and the required sample size was confirmed to be around 3–5,000 or more . We estimated the priors, π and g, based on the data by applying an expectation–maximization (EM) algorithm, that is, empirical Bayes estimation. The non-parametric estimate of g was discrete with mass points p =  at a series of nonzero points b =  (b1 < b2 < ··· < bB). We set b1 = −0.3 and bB = 0.3 (0.74 and 1.35 in odds ratio). The number grid point B = 120 was used, such that b = . The initial value of π and the initial distribution of g were important and determined by a careful procedure , is given byFor the μ, 1(·) denotes the cumulative distribution function of the normal distribution with mean μ and unit variance, and zc denotes the rejection threshold determined by a significance level, pc, satisfying pc = 5 × 10−8 (the genome-wide “significant” threshold) and pc = 1 × 10−6 (the genome-wide “suggestive” threshold) were used. Under the SP-HMM, the rejection probability, i.e., the probability that the j-th SNP is significant, is given bywhere Φnr and ns be the sample sizes for cases and controls, respectively, in an existing GWAS from which we can estimate the SP-HMM. In addition, we envisage a “future” GWAS with j-th SNP in the future GWAS can be obtained through replacing nr + 1/ns) ×  × . We set m* = 100,000 for targeting 100,000 pruned SNPs. Since the term  with the selected SNP removed. This was repeated until no SNPs remained. LD information(r2) was extracted from the HapMap database  . The quality control and pruning for the SNP data were done as described in the previous subsection, “GWAS Data.” For SCZ, bipolar disorder, and coronary artery disease, there were 101314, 96681, and 79512 SNPs in the pruned sets, respectively. Those values were set as m* in the formula (2). The number of SNPs was smaller for coronary artery disease (79512), as the original GWAS summary data have been imputed using HapMap data. Table We validated our approach for predicting the number of significant SNPs using hypothetical “current” and “future” GWAS data; we fitted the SP-HMM to the “current” GWAS data with smaller sample size to predict the number of significant SNPs in the “future” GWAS data with larger sample size, and we compared the predicted value with the observed one. The three pairs of GWAS summary statistics for SCZ (for “current” data, Cross-Disorder Group of the Psychiatric Genomics Consortium, JN and SM: Conceptualization; JN: Formal analysis; TT and SM: Funding acquisition; JN and SM: Writing original draft; HO, YK and TT: Writing review and editing.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Inflammatory activity of MECN was assessed by carrageenan-induced paw oedema while analgesic activity was evaluated by acetic acid –induced writhing and formalin paw lick test. Histological analyses of the paws were also carried out. There was evaluation of the mechanism(s) of action of MECN using naloxone, a blocker of opioid receptors; atropine, blocker of muscarinic receptors; and propranolol, blocker of beta adrenergic receptors. Findings from the study revealed that MECN has both anti-inflammatory and analgesic properties. These properties were found to be dose dependent with 200 mg/kg of MECN discovered to be the most potent dose. 200 mg/kg was able to cause statistically significant reduction in paw size (p < 0.001) when compared with the carrageenan group. Histological analysis revealed that rats treated with 200 mg/kg of MECN showed no inflammatory cells in the left paw compared to other groups treated with carrageenan. In the formalin test, the number of paw licking was significantly reduced by MECN at 50 mg/kg, 100 mg/kg and 200 mg/kg in both neurogenic and inflammatory pain responses (p < 0.001) even as 200 mg/kg showed the highest percentage inhibition of 98.17% while 100 mg/kg of aspirin showed percentage inhibition of 93.66%. In acetic acid-induced writhing test, 50 mg/kg, 100 mg/kg and 200 mg/kg of MECN produced significant inhibition of writhes when compared with control as highest inhibition is observed in mice that received 200 mg/kg which is similar to aspirin. Administration of propranolol and naloxone was unable to reverse the analgesic function of MECN. However, atropine administration blocked the analgesic function of MECN. This shows that MECN exhibits its analgesic property through cholinergic pathway and not opioid and adrenergic pathways. Phytochemical screening revealed that MECN contains flavonoids, steroids, saponins, tannins, anthraquinines, terpenoids, and alkanoids. These phytochemical contents may thus be responsible for its analgesic and anti-inflammatory properties.Methanol extract of  Glacial acetic acid, formaldehyde, carrageenan, were purchased from S.D. Fine Chemical Pvt. Ltd. . The extract was dissolved in tween 20. All other chemical were of analytical graded. Aspirin, ibuprofen, naloxone hydrochloride, atropine sulphate, and propranolol were purchased from Alpha Pharmacy, Lagos, Nigeria.2.2ad libitum. The animals were kept in separate cages according to their groups. Animals were deprived of food for 16hrs before experimentation in analgesic acid model to prevent interaction of food with the visceral writhing test. All procedures involving the use of animals in this study complied with the guiding principles for research involving animals as recommended by the declaration of Helsinki and the Guiding principles in the care and use of animals.Female Wistar rats weighing 100–120 g and aged 8–10 weeks and Albino mice (15–23 g) aged 10–12 weeks were acclimatized for two weeks before the initiation of the experiment, and maintained under standard nutritional and environmental condition (12-hour light/day cycle). They were purchased from Animal House, Bowen University, Iwo. They had free access to standard pellet diet and water 2.3Cola nitida were collected in Odori market, Iwo. The identification and authentication of the seeds was carried out at Department of Botany, Obafemi Awolowo University, Ile-Ife, Nigeria on the 16th March 2016 with voucher number: FHI.17512.Fresh seeds of 2.4Cola nitida powder was soaked in several beakers, in 8.365 L of methanol for 72 h at room temperature after which it was filtered using 150 mm Whatmann filter paper then the residue was squeezed to yield more filtrate. It was then evaporated to dryness by a rotary evaporator at 40 °C. Percentage yield of Cola nitida was 3.79%. The yield weighed 240 g.The seeds were air dried at room temperature and ground to fine powder. It was then extracted, using gradient extraction method. 3.56 kg of grinded 2.5Cola nitida  was administered orally one hour before 0.1 ml of 1% carrageenan was delivered into the sub-plantar surface of the left hind paw of the rats in the five groups. The extract and ibuprofen were administered using Tween 20 and normal saline as vehicle. The paw sizes were measured at 0, 1, 2, 3, 4, 5, 6 h, and also 24 h after injection of carrageenan. After the 24 h of measuring the paw size for each group, the rats were then euthanized and the paw tissues were harvested for histological evaluationBefore the experiment began, the paw sizes of the left hind paw of the rats were determined using the automatic vernier caliper. The rats were then divided into five groups containing six animals each. Normal Saline 10 ml/kg, ibuprofen 10 mg/kg , or Cola2.62.6.1Cola nitida extract respectively through oral administration. Group 5 received 100 mg/kg of aspirin orally . 2 mg/kg doses were used for these antagonist drugs as obtained from similar studies •Cola nitida (200 mg/kg)Group II was administered with •Cola nitida (200 mg/kg)Group III was administered with Distilled Water and •Group IV was administered with Aspirin (100 mg/kg)•Cola nitida (200 mg/kg)Group V was administered with Naloxone (2 mg/kg) and For Naloxone;•Group I was administered with Distilled Water (10 ml/kg)•Cola nitida (200 mg/kg)Group II was administered with •Cola nitida (200 mg/kg)Group III was administered with Distilled Water and •Group IV was administered with Aspirin (100 mg/kg)•Cola nitida (200 mg/kg)Group V was administered with Atropine (2 mg/kg) and of For Atropine;•Group I was administered with Distilled Water (10 ml/kg)•Cola nitida (200 mg/kg)Group II was administered with •Cola nitida (200 mg/kg)Group III was administered with Distilled Water and •Group IV was administered with Aspirin (100 mg/kg)•Cola nitida (200 mg/kg)Group V was administered with Propranolol (3 mg/kg) and For Propranolol;2.8Cola nitida was subjected to preliminary screening for various active phytochemical constituents such as alkaloids, cardenoldes, anthraquinone, saponins, flavonoids and tannins. The phytochemical screening can be carried out using Drangenduff’s reagent, Meyer’s reagent or Wanger’s reagent for alkaloids; keller-killiani reagent or kedde reagent for cardenoldes; chloroform/ammonia test for free anthraquinone; frothing test for saponins; ferric chloride test for tannins and DPPH for flavonoids . The slides were examined under a light microscope with the photomicrographs taken.2.10Post-hoc testing was performed for inter-group comparison using the Bonferroni test. Results were considered to be significant at p < 0.05.Data were expressed as mean ± standard error of the mean. The data obtained were analysed using one-way ANOVA by Graphpad Prism 7.0. 33.13.23.33.43.5Cola nitida extract. There was significant reduction (P < 0.0001) in paw licking time in both early (0–5 mins) and late phase (20–30 mins) in groups 2, 3, and 4 treated with 200 mg/kg compared with the control group that received acetic acid only. Group 4 pretreated with naloxone also showed statistically significant reduction (P < 0.0001) in paw licking time in both early and late phase when compared with the control group treated with acetic acid only. This outcome shows that the mechanism of anti-nociceptive action of MECN is not through the opioid receptors.3.6Cola nitida extract. There was significant reduction (p < 0.0001) in paw licking time in both early (0–5 mins) and late phase (20–30 mins) in the all the groups treated with 200 mg/kg MECN compared with the control group that received only distilled water. Group 5 pretreated with atropine however showed no statistically significant reduction (P < 0.05) in paw licking time in both early and late phase when compared with the control group. This result suggested that cholinergic pathway plays a significant role in mediating the anti-nociceptive action of Cola nitida.3.7Cola nitida does not exert its anti-nociceptive action through the beta-adrenergic pathway  in paw licking time in both early and late phase. Group 4 pretreated with propranolol also showed statistically significant reduction (P < 0.05) in paw licking time in both early and late phase when compared with the control group. The result showed that  pathway .Table 5E4Cola nitida. The findings from the study revealed that methanol extract of Cola nitida possesses anti-inflammatory and anti-nociceptive properties. These properties were found to be dose dependent with 200 mg/kg of MECN discovered to be the most potent dose. Also, the phytochemical screening revealed that MECN contains flavonoids, steroids, saponins, tannins, anthraquinines, terpenoids, and alkanoids. This is similar to the findings of The aim of the study is to investigate the anti-nociceptive and anti-inflammatory properties of Carrageenan induced inflammation has been used for inflammatory studies to test the potency of anti-inflammatory agents . AlthougCarrageenan injection produces paw oedema in a biphasic fashion. Oedema in the first phase is mediated by histamine, serotonin and bradykinin while the second phase (3–5 h) is mediated by prostaglandins . 50 mg/kFormalin test has been used as a model for localized inflammatory pain and tonic pain . There aAcetic acid-induced writhing test is also a model that had been used to test the potency of analgesic agents . The tesWe proceeded further to investigate the mechanism of action of the analgesic property of MECN. Naloxone and propranolol are non-selective opioid receptors blocker and selective beta adrenergic blocker respectively that have been used in studies that evaluate mechanisms of action of analgesic compounds . PropranThere are several studies proving that flavonoids in plant extracts have anti-inflammatory and analgesic effects . It has 5MECN was found to have analgesic and anti-inflammatory properties in a dose dependent manner. 200 mg/kg of MECN gave a better performance than aspirin in formalin-induced paw licking test. The analgesic effect of MECN seems to be mediated through cholinergic pathway.Authors have declared that no competing interest exist.