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Liquid chromatography paired with tandem mass spectrometry (LC-MS/MS) is the gold standard in measurement of endocannabinoid concentrations in biomatrices. We conducted a systematic review of literature to identify advances in targeted LC-MS/MS methods in the period 2017-2024. We found that LC-MS/MS methods for endocannabinoid quantification are relatively consistent both across time and across biomatrices. Recent advances have primarily been in three areas: (1) sample preparation techniques, specific to the chosen biomatrix; (2) the range of biomatrices tested, recently favoring blood matrices; and (3) the breadth of endocannabinoid and endocannabinoid-like analytes incorporated into assays. This review provides a summary of the recent literature and a guide for researchers looking to establish the best methods for quantifying endocannabinoids in a range of biomatrices.
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Linoleic acid (LA) is required for neuronal development. We have previously demonstrated sex-specific changes in cardiovascular and hepatic function in rat offspring from mothers consuming a high-LA diet, with some effects associated with reduced LA concentration in the postnatal diet. At this time, the impact of a high-maternal-LA diet on offspring brain development and the potential for the postnatal diet to alter any adverse changes are unknown. Rat offspring from mothers fed low- (LLA) or high-LA (HLA) diets during pregnancy and lactation were weaned at postnatal day 25 (PN25) and fed LLA or HLA diets until sacrifice in adulthood (PN180). In the offspring's brains, the postnatal HLA diet increased docosapentaenoate in males. The maternal HLA diet increased LA, arachidonate, docosapentaenoate, C18:0 dimethylacetal (DMA), C16:0 DMA, C16:0 DMA/C16:0, and C18:0 DMA/C18:0, but decreased eoicosenoate, nervoniate, lignocerate, and oleate in males. Maternal and postnatal HLA diets reduced oleate and vaccenate and had an interaction effect on myristate, palmitoleate, and eicosapentaenoate in males. In females, maternal HLA diet increased eicosadienoate. Postnatal HLA diet increased stearate and docosapentaenoate. Maternal and postnatal HLA diets had an interaction effect on oleate, arachidate, and docosahexaenoic acid (DHA)/omega (n)-6 docosapentaenoic acid (DPA) in females. Postnatal HLA diet decreased DHA/n-6 DPA in males and females. Postnatal HLA diet increased plasma endocannabinoids (arachidonoyl ethanolamide and 2-arachidonoyl glycerol), as well as other N-acyl ethanolamides and testosterone. HLA diet alters brain fatty acids, plasma endocannabinoids, and plasmalogen concentrations in a development-specific and sex-specific manner.
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Encéfalo , Endocanabinoides , Ácidos Graxos , Ácido Linoleico , Plasmalogênios , Feminino , Animais , Masculino , Gravidez , Ratos , Encéfalo/metabolismo , Ácidos Graxos/sangue , Ácidos Graxos/metabolismo , Endocanabinoides/sangue , Endocanabinoides/metabolismo , Ácido Linoleico/sangue , Plasmalogênios/sangue , Plasmalogênios/metabolismo , Efeitos Tardios da Exposição Pré-Natal/sangue , Caracteres Sexuais , Fatores SexuaisRESUMO
The replicability of fear conditioning research has come under recent scrutiny, with increasing acknowledgment that the use of differing materials and methods may lead to incongruent results. Direct comparisons between the main two unconditional stimuli used in fear conditioning - an electric shock or a loud scream-are scarce, and yet these stimuli are usually used interchangeably. In the present study, we tested whether a scream, a shock, or an unpredictable combination of the two affected fear acquisition, extinction, and return of fear amongst healthy participants (N = 109, 81 female). We also collected hair samples and tested the relationship between fear conditioning and hair endocannabinoid levels. Our findings suggest that, although subjective ratings of pleasantness, arousal, and anxiety were similar regardless of the unconditional stimuli used, skin conductance responses were significantly lower for stimuli paired with the scream compared to a shock alone. Further, reducing the predictability of the unconditional stimulus reduced habituation of skin conductance responses during acquisition and reacquisition, but did not produce stronger conditioning compared to shock alone. Exploratory analyses suggested that hair endocannabinoids were associated with overall physiological arousal during fear conditioning, as well as higher return of fear to the threat cue, but not to the safety cue. These findings have multiple implications for the design and replicability of fear conditioning research and provide the first evidence for an association between hair levels of endocannabinoids and human fear conditioning.
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Endocanabinoides , Extinção Psicológica , Humanos , Feminino , Extinção Psicológica/fisiologia , Resposta Galvânica da Pele , Condicionamento Clássico/fisiologia , Medo/fisiologia , CabeloRESUMO
BACKGROUND: Transcranial direct current stimulation (tDCS) has previously been shown to improve fear extinction learning and retention when administered prior to or during extinction learning. This study investigates whether tDCS immediately following extinction learning improves efficacy of extinction memory retention. METHODS: 30 participants completed a 2-day fear learning and extinction paradigm, where they acquired fear of a stimulus conditioned to an aversive electric shock on day 1. Extinction learning occurred on day 1, with tDCS or sham tDCS administered immediately following the learning phase. Participants returned for a second day test of extinction memory recall. Skin conductance was measured as the primary outcome. RESULTS/CONCLUSIONS: Participants in the tDCS group showed impaired fear extinction retention on day 2, marked by significant generalisation of fear to the safety stimulus. This contrasts with earlier studies showing improved extinction retention when stimulation occurred during encoding of extinction learning, compared to immediate consolidation as in our study. These findings may have important implications for the use of tDCS during exposure therapy for anxiety and trauma disorders.
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Extinção Psicológica , Medo/fisiologia , Retenção Psicológica , Estimulação Transcraniana por Corrente Contínua , Adulto , Extinção Psicológica/fisiologia , Medo/psicologia , Feminino , Resposta Galvânica da Pele , Humanos , Masculino , Retenção Psicológica/fisiologia , Fatores de Tempo , Estimulação Transcraniana por Corrente Contínua/métodos , Adulto JovemRESUMO
BACKGROUND: The endocannabinoid system is gaining increasing attention as a favorable target for improving posttraumatic stress disorder (PTSD) treatments. Exposure therapy is the gold-standard treatment for PTSD, and fear extinction learning is a key concept underlying successful exposure. METHODS: This study examined the role of genetic endocannabinoid polymorphisms in a fear extinction paradigm with PTSD compared to healthy participants (N = 220). Participants provided saliva for genotyping, completed a fear conditioning and extinction task, with blood samples taken before and after the task (n = 57). Skin conductance was the outcome and was analyzed using mixed models. RESULTS: Results for cannabinoid receptor type 1 polymorphisms suggested that minor alleles of rs2180619 and rs1049353 were associated with poorer extinction learning in PTSD participants. The minor allele of the fatty acid amide hydrolase (FAAH) polymorphism rs324420 was associated with worse extinction in PTSD participants. Subanalysis of healthy participants (n = 57) showed the FAAH rs324420 genotype effect was dependent on plasma arachidonoyl ethanolamide (AEA) level, but not oleoylethanolamide or 2-arachidonoyl glycerol. Specifically, higher but not lower AEA levels in conjunction with the minor allele of FAAH rs324420 were associated with better extinction learning. CONCLUSIONS: These findings provide translational evidence that cannabinoid receptor 1 and AEA are involved in extinction learning in humans. FAAH rs324420's effect on fear extinction is moderated by AEA plasma level in healthy controls. These findings imply that FAAH inhibitors may be effective for targeting anxiety in PTSD, but this effect needs to be explored further in clinical populations.
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Canabinoides , Endocanabinoides , Extinção Psicológica , Medo , Humanos , AprendizagemRESUMO
Background: PTSD is characterised by severe sleep disturbances, which is increasingly recognised to in many cases consist of similar symptomology to sleep disorders such as REM Behaviour Disorder (RBD). The present study aimed to investigate whether different aspects of sleep quality influence intrusive memory development and whether PTSD status moderates this relationship. Participants and Methods: 34 PTSD, 52 trauma-exposed (TE) and 42 non-trauma exposed (NTE) participants completed an emotional memory task, where they viewed 60 images (20 positive, 20 negative and 20 neutral) and, two days later, reported how many intrusive memories they had of each valence category. Participants also completed three measures of sleep quality: the Pittsburgh Sleep Quality Index, the REM Behaviour Disorder Screening Questionnaire and total hours slept before each session. Results: The PTSD group reported poorer sleep quality than both TE and NTE groups on all three measures, and significantly more negative intrusive memories than the NTE group. Mediation analyses revealed that self-reported RBD symptomology before the second session mediated the relationship between PTSD status and intrusive memories. Follow-up moderation analyses revealed that self-reported RBD symptomology before the second session was only a significant predictor of intrusion in the PTSD group, though with a small effect size. Conclusions: These findings suggest that RBD symptomology is an indicator of consolidation of intrusive memories in PTSD but not trauma-exposed or healthy participants, which supports the relevance of characterising RBD in PTSD.
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Rememoração Mental , Transtorno do Comportamento do Sono REM/psicologia , Autorrelato , Transtornos de Estresse Pós-Traumáticos/psicologia , Adulto , Emoções , Humanos , Masculino , Sono , Adulto JovemRESUMO
Immunoassay overestimates progesterone in blood, but no studies have tested whether this occurs in saliva. We measured progesterone in saliva using immunoassay and mass spectrometry. We tested the immunoassay for cross reactivity with dehydroepiandrosterone sulfate (DHEA-S) and 17α-hydroxyprogesterone (17α-OHP). Progesterone was significantly higher in immunoassay compared to mass spectrometry. Immunoassay progesterone levels increased in when incremental levels of 17α-OHP standard was added. This effect was not observed with the addition of DHEA-S. Research using salivary progesterone immunoassay techniques should be wary, particularly with individuals taking steroid supplementation or with high levels of progesterone metabolites.
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Imunoensaio , Progesterona/análise , Saliva/química , Espectrometria de Massas em Tandem , 17-alfa-Hidroxiprogesterona/análise , 17-alfa-Hidroxiprogesterona/normas , Adulto , Cromatografia Líquida de Alta Pressão , Sulfato de Desidroepiandrosterona/análise , Sulfato de Desidroepiandrosterona/normas , Feminino , Humanos , Limite de Detecção , Masculino , Pessoa de Meia-Idade , Padrões de Referência , Reprodutibilidade dos Testes , Adulto JovemRESUMO
Renewal is a 'return of fear' manipulation in human fear conditioning to investigate learning processes underlying anxiety and trauma. Even though renewal paradigms are widely used, no study has compared the strength of different renewal paradigms. We conduct a systematic review (N = 80) and meta-analysis (N = 23) of human fear conditioning studies assessing renewal. Our analysis shows that the classic ABA design is the most effective paradigm, compared to ABC and ABBA designs. We present evidence that conducting extinction in multiple contexts and increasing the similarity between acquisition and extinction contexts reduce renewal. Furthermore, we show that additional cues can be used as safety and 'protection from extinction' cues. The review shows that alcohol weakens the extinction process and that older adults appear less sensitive to context changes and thus show less renewal. The large variability in approaches to study renewal in humans suggests that standardisation of fear conditioning procedures across laboratories would be of great benefit to the field.
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Extinção Psicológica , Medo , Medo/fisiologia , Humanos , Extinção Psicológica/fisiologia , Condicionamento Clássico/fisiologia , Sinais (Psicologia) , Condicionamento Psicológico/fisiologiaRESUMO
The endogenous cannabinoid (ECB) system is a small molecule lipid signalling system that is involved in stress response activation and is associated with PTSD, but it is unclear whether salivary ECBs are part of the sympathetic nervous system response to stress. We conducted an adapted trauma film paradigm, where participants completed a cold pressor test (or control) while watching a 10-minute trauma film. We also collected saliva and hair samples and tested them for ECBs, cortisol, and salivary alpha amylase (sAA). As hypothesised, there were significant positive correlations between sAA activity and salivary ECB levels, particularly 2-arachidonoyl glycerol (2-AG), though ECBs were not correlated with sAA stress reactivity. Participants who had a significant cortisol response to the trauma film/stressor reported less intrusive memories, which were also less distressing and less vivid. This effect was moderated by arachidonoyl ethanolamide (AEA), where decreases in AEA post-stress were associated with more intrusive memories in cortisol non-responders only. This study provides new evidence for the role of ECBs in the sympathetic nervous system.
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Ácidos Araquidônicos , Hidrocortisona , alfa-Amilases Salivares , Humanos , Endocanabinoides , Alcamidas Poli-Insaturadas , SalivaRESUMO
Presenting unpaired unconditional stimuli (US) during extinction training reduces the renewal of conditional fear due to context change and slows re-acquisition. The present study investigated whether this reduced return of fear is mediated by Pavlovian inhibitory conditioning to the conditional stimulus paired with the US during acquisition (CS+) that is acquired when this stimulus is presented without the US in an excitatory extinction context. Using an ABA renewal paradigm that trained extinction in a context different from acquisition and renewal test, participants either received no USs (Standard), five unsignalled US presentations (Unsignalled) or five presentations of the US preceded by a novel, third CS (Signalled) during extinction training. Extinction was followed by tests for renewal and re-acquisition. Replicating previous results, renewal of electrodermal conditional responses was observed in group Standard, but not in group Unsignalled. Signalling the additional USs, and thus reducing context conditioning and the potential for inhibitory conditioning, did not reduce their effect in that renewal was absent in group Signalled. These results are inconsistent with an inhibitory conditioning account of the effects of unpaired US presentations during extinction. A trial sequence learning account or an arousal account may explain the effects of unpaired presentations of the US during extinction.
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The emotional conflict task measures emotional conflict resolution and adaptation, but some studies are unable to find resolution or adaptation effects using this task. We examined boundary conditions and replicability of the emotional conflict resolution and adaptation effects through secondary data analysis, systematic review, and meta-analysis of studies in the field. In our data, we were unable to fully replicate the emotional conflict resolution or adaptation effects and found that most studies using this task (n = 94) do not report analysis of emotional conflict resolution, with only 28% (n = 26) studies doing so. Our meta-analysis suggests that studies reporting emotional conflict resolution and adaptation analyses overall report significant but small effects, suggesting the effect is difficult to consistently replicate. Our meta-analysis revealed that controlling for contingency learning may impact the ability of studies to identify conflict resolution. These findings have implications for assessment and interpretation of the emotional conflict task. (PsycInfo Database Record (c) 2024 APA, all rights reserved).
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Adaptação Psicológica , Conflito Psicológico , Emoções , Humanos , Emoções/fisiologia , Reprodutibilidade dos Testes , Negociação/psicologia , Análise de Dados SecundáriosRESUMO
Meta-analyses indicate differences in Pavlovian fear responses between anxious and non-anxious individuals using electrodermal activity (EDA). Recent research, however, has cast doubt on whether these effects are robust to different analytic choices. Using the multiverse approach conceived by Steegen et al. (2016), we surveyed analytic choices typically implemented in clinical fear conditioning research by conducting 1240 analyses reflecting different choice permutations. Only 1.45% of our analyses produced theoretically congruent statistically significant effects, and the strength and direction of the estimated effects varied substantially across EDA processing methods. We conclude that EDA-estimated fear learning differences are vulnerable to researcher degrees of freedom and make recommendations regarding which analytical choices should be approached with a high degree of caution.
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Recent studies have shown that fear conditioning experiments can be successfully conducted online. However, there is limited evidence that measures other than subjective ratings of threat expectancy can be collected, which means that online research may not be able to adequately replace laboratory experiments. In the current study, we conducted an online fear conditioning experiment consisting of habituation, acquisition, extinction and 48 h delayed extinction recall using ratings of threat expectancy and conditional stimulus pleasantness, and probe reaction time as outcome measures. The conditional stimuli were categories of words and a levels of processing manipulation explored whether words that were processed at a deeper level during extinction evoked smaller differential threat responses during extinction recall. Although the levels of processing manipulation did not produce a significant outcome, we found that extinction recall was successfully operationalised in our study. Reaction time indicated differential responding during both acquisition and extinction recall, and age of participants was correlated in one of two experiments with differential threat expectancy and reaction time, such that older participants showed better safety learning. The outcomes of this experiment provide multiple novel tools for researchers to explore fear conditioning, especially in an online environment.
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Extinção Psicológica , Medo , Humanos , Tempo de Reação , Extinção Psicológica/fisiologia , Emoções/fisiologia , Resposta Galvânica da Pele , Avaliação de Resultados em Cuidados de SaúdeRESUMO
Fear conditioning is a significant area of research that has featured prominently among the topics published in Biological Psychology over the last 50 years. This work has greatly contributed to our understanding of human anxiety and stressor-related disorders. While mainly conducted in the laboratory, recently, there have been initial attempts to conduct fear conditioning experiments online, with around 10 studies published on the subject, primarily in the last two years. These studies have demonstrated the potential of online fear conditioning research, although challenges to ensure that this research meets the same methodological standards as in-person experimentation remain, despite recent progress. We expect that in the coming years new outcome measures will become available online including the measurement of eye-tracking, pupillometry and probe reaction time and that compliance monitoring will be improved. This exciting new approach opens new possibilities for large-scale data collection among hard-to-reach populations and has the potential to transform the future of fear conditioning research.
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Condicionamento Clássico , Medo , Humanos , Medo/psicologia , Ansiedade/psicologia , Transtornos de Ansiedade , Tempo de ReaçãoRESUMO
On the basis of substantial preclinical evidence, the endogenous cannabinoid system has been proposed to be closely involved in stress reactivity and extinction of fear. Existing human research supports this proposal to some extent, but existing studies have used only a narrow range of tools and biomatrices to measure endocannabinoids during stress and fear experiments. In the present study we collected hair and saliva samples from 99 healthy participants who completed a fear conditioning and intrusive memory task. Subjective, physiological and biological stress reactivity to a trauma film, which later served as unconditional stimulus during fear conditioning, was also measured. We found that salivary endocannabinoid concentrations predicted subjective responses to stress, but not cortisol stress reactivity, and replicated previous findings demonstrating a sex dimorphism in hair and salivary endocannabinoid levels. Hair 2-arachidonoyl glycerol levels were significantly associated with better retention of safety learning during extinction and renewal phases of fear conditioning, while hair concentrations of oleoylethanolamide and palmitoylethanolamide were associated with overall physiological arousal, but not conditional learning, during fear conditioning. This study is the first to test the relationship between hair and salivary endocannabinoids and these important psychological processes. Our results suggest that these measures may serve as biomarkers of dysregulation in human fear memory and stress.
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Endocanabinoides , Extinção Psicológica , Humanos , Extinção Psicológica/fisiologia , Condicionamento Clássico/fisiologia , Medo/fisiologia , CabeloRESUMO
INTRODUCTION: Preclinical and experimental research have provided promising evidence that medicinal cannabis may be efficacious in the treatment of posttraumatic stress disorder (PTSD). However, implementation of medicinal cannabis into routine clinical therapies may not be straightforward. AREAS COVERED: In this review, we describe some of the clinical, practical, and safety challenges that must be addressed for cannabis-based treatment of PTSD to be feasible in a real-world setting. These issues are especially prevalent if medicinal cannabis is to be combined with trauma-focused psychotherapy. EXPERT OPINION: Future consideration of the clinical and practical considerations of cannabis use in PTSD therapy will be essential to both the efficacy and safety of the treatment protocols that are being developed. These issues include dose timing and titration, potential for addiction, product formulation, windows of intervention, and route of administration. In particular, exposure therapy for PTSD involves recall of intense emotions, and the interaction between cannabis use and reliving of trauma memories must be explored in terms of patient safety and impact on therapeutic outcomes.
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Cannabis , Alucinógenos , Maconha Medicinal , Transtornos de Estresse Pós-Traumáticos , Humanos , Transtornos de Estresse Pós-Traumáticos/tratamento farmacológico , Transtornos de Estresse Pós-Traumáticos/psicologia , Maconha Medicinal/efeitos adversos , Austrália , Psicoterapia/métodos , Alucinógenos/uso terapêuticoRESUMO
Dysregulated consolidation of emotional memories is a core feature of posttraumatic stress disorder (PTSD). Brain Derived Neurotrophic Factor (BDNF) influences synaptic plasticity and emotional memory consolidation. The BDNF Val66Met polymorphism has been associated with PTSD risk and memory deficits respectively, although findings have been inconsistent, potentially due to a failure to control for important confounds such as sex, ethnicity, and the timing/extent of previous trauma experiences. Furthermore, very little research has examined the impact of BDNF genotypes on emotional memory in PTSD populations. This study investigated the interaction effects of Val66Met and PTSD symptomatology in an emotional recognition memory task in 234 participants divided into healthy control (n = 85), trauma exposed (TE: n = 105) and PTSD (n = 44) groups. Key findings revealed impaired negative recognition memory in PTSD compared to control and TE groups and in participants with the Val/Met compared to the Val/Val genotype. There was a group × genotype interaction showing no Met effect in the TE group despite significant effects in PTSD and controls. Results suggest that people previously exposed to trauma who do not develop PTSD may be protected from the BDNF Met effect, however more research is needed to replicate findings and to explore the epigenetic and neural processes involved.
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Transtornos de Estresse Pós-Traumáticos , Humanos , Fator Neurotrófico Derivado do Encéfalo/genética , Emoções , Genótipo , Transtornos da Memória , Polimorfismo de Nucleotídeo Único , Transtornos de Estresse Pós-Traumáticos/genéticaRESUMO
The endocannabinoid (ECB) system has recently been considered a potential treatment target for various clinical disorders. However, research around age- and sex-related changes within the ECB system is relatively limited. To improve our understanding of these changes, the current study measured arachidonoyl ethanolamide (AEA), 2-arachidonoyl glycerol (2-AG), oleoylethanolamine (OEA), palmitoylethanolamine (PEA), arachidonic acid (AA), cortisol, and progesterone in pooled serum samples stratified by sex (male and female) and age groups (5-15; 15-30; 30-45; 45-60; 60-75; 85+), using liquid-chromatography tandem mass spectrometry. Serum progesterone levels significantly increased in females of the 15-30 and 30-45 age groups, before declining. Significantly higher cortisol, AEA, 2-AG, OEA, and PEA were found in males and in older age, while significantly higher AA was found in females. Our results indicate that ECBs and related hormones exhibit sexual dimorphism in the age ranges that correspond with female pregnancy, menopause, and post menopause. Male testosterone levels most likely influences male ECB changes throughout the lifespan. Future research could capitalise on these findings by performing repeated measurements in individuals in a longitudinal style, to further refine the temporal profile of age-specific changes to the ECB system identified here.
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Endocanabinoides , Caracteres Sexuais , Gravidez , Humanos , Masculino , Feminino , Longevidade , Progesterona , Hidrocortisona , EtanolaminasRESUMO
A growing literature has sought to combine fear conditioning paradigms with the trauma film paradigm to study the associative learning properties of intrusive re-experiencing in PTSD. We review this innovative approach and the recent findings by highlighting their relevance to cognitive and conditioning theories of PTSD. We also conduct a meta-analysis of the available studies to demonstrate that, for most outcome measures, fear learning using a traumatic film clip unconditional stimulus yields results similar to those seen with an electro-tactile unconditional stimulus, which implies that the combined paradigm shares at least some properties of more standard fear conditioning paradigms. We argue that careful use of this combined paradigm will provide important new insights into the mechanisms underlying memory symptoms of PTSD and will allow rigorous testing of cognitive theories of the disorder.