[Conditioning of the immune system-Already clinically usable?] / Konditionierung des Immunsystems ­ Schon klinisch nutzbar?

The brain and the immune system permanently exchange information via various neuronal and humoral signaling pathways. This communication network forms the basis for controlling peripheral immune functions via associative learning or conditioning processes. Establishing a learned immune reaction, an immunomodulatory drug that represents the unconditioned stimulus (US) is paired with a new odor or taste stimulus. Re-presentating this previously neutral odor or taste stimulus, its now functions as a conditioned stimulus (CS) and triggers reactions in the immune system similar to those formerly induced by the drug used as US. Using different learning protocols, it was possible to condition immunopharmacological effects in animal disease models, such as lupus erythematosus, contact allergy or rheumatoid arthritis, thereby reducing disease symptoms. Preliminary experimental studies in healthy volunteers and patients confirmed a possible clinical use of learned immune responses with the aim of using associative learning protocols as complementary measures to pharmacological interventions in clinical practice in order to reduce drug doses and thus undesirable drug side effects while maintaining therapeutic efficacy. However, there is still a great need for further research to understand the mechanisms of learned immune responses in preclinical studies and to optimize the associative learning processes for using them in the clinical routine in studies with healthy volunteers and patients.

[The effect of treatment expectations on pruritus and skin pain]. / Einfluss der Behandlungserwartung auf Pruritus und Hautschmerzen.

BACKGROUND: Patients' expectations in terms of the benefit of a treatment are key determinants of placebo responses and can affect the development and course of medical conditions as well as the efficacy and tolerability of active medical treatment. The mechanisms mediating these placebo and nocebo effects have been best described in the field of experimental pain and placebo analgesia. However, also in dermatology experimental and clinical studies demonstrate that different skin symptoms such as itch, skin pain and dermatologic diseases can be modulated by patients' expectations. OBJECTIVES: The aim of this review is to provide a current overview of the empirical evidence for the effects of patients' expectations in the field of dermatology with a focus on different skin symptoms such as itch and pain. Finally, the relevance of this topic for physicians who treat patients with dermatologic symptoms is discussed. MATERIALS AND METHODS: The article is a narrative review. RESULTS: Steadily growing evidence from experimental and clinical studies in healthy volunteers and dermatologic patients suggests that patients' positive treatment expectations can reduce skin disease symptoms and enhance treatment efficacy, while negative treatment expectations can induce a nocebo effect associated with increased symptomatology. Patients' prior treatment experiences as well as the quality and quantity of doctor-patient communication play a central role in shaping treatment expectations. CONCLUSIONS: Techniques aimed at maximizing positive expectation effects in patients should be implemented in daily clinical routine.

[The learned placebo response in the immune system]. / Die gelernte Placeboantwort im Immunsystem.

The learned placebo response of the immune system is based on the mutual interaction between the brain and the immune system; both systems continually exchange information via humoral and neural communication pathways. This communication network enables the modification, i.e. suppression or stimulation, of peripheral immune functions by classical or Pavlov's conditioning. The present article provides an overview of the results of recent experimental animal studies, which also document the potential clinical relevance of learned immune responses. Learned immunological responses mediated by classical conditioning have also been demonstrated in humans. The knowledge gained from experimental data and clinical observations paves the way for a potential implementation of learned immune responses as supportive measures to standard immunopharmacological treatment strategies to reduce drug dosage as well as adverse side effects while simultaneously maximizing the therapeutic effect.

Immunological effects of behavioral activation with exercise in major depression: an exploratory randomized controlled trial.

Major depression (MD) is associated with peripheral inflammation and increased cardiovascular risk. Regular physical exercise can have anti-inflammatory effects. The present study examined whether behavioral activation with exercise affects inflammatory processes in MD. Ninety-eight patients with MD were randomly assigned to cognitive-behavioral therapy (CBT) emphasizing exercise during behavioral activation (CBT-E), CBT with pleasurable low-energy activities as an active control condition (CBT-C) or a passive waiting list control group (WL). Plasma levels of C-reactive protein (CRP), interleukin (IL)-6, IL-10, lipopolysaccharide (LPS)-stimulated IL-6 production, and blood immune cell counts were analyzed at baseline and weeks 8 (post-behavioral activation) and 16 (post-treatment). Thirty non-depressed age- and sex-matched controls were included to examine potential immunological alterations in MD at baseline. Patients with MD exhibited higher levels of CRP, higher neutrophil and monocyte counts, lower IL-10 levels and reduced LPS-stimulated IL-6 production compared to controls (P<0.001-0.045). Multilevel modeling indicated that CBT-E was associated with increased anti-inflammatory IL-10 at weeks 8 and 16 compared to CBT-C and WL (P=0.004-0.018). CBT-E did not significantly affect other immunological makers in the total sample. A subgroup analysis including patients with potentially higher cardiovascular risk (CRP ⩾1 µg ml-1) indicated that CRP was reduced in CBT-E compared to CBT-C (P<0.007) and marginally reduced compared to WL (P<0.085) after week 16. The present findings provide new insights into immunological effects of behavioral treatments against depression. Behavioral activation in conjunction with exercise may have the potential to reverse, in part, immunological alterations in MD.

Selective increase of cerebrospinal fluid IL-6 during experimental systemic inflammation in humans: association with depressive symptoms.

Systemic inflammation is accompanied by profound behavioral and mood changes that resemble symptoms of depression. Findings in animals suggest that pro-inflammatory cytokines released by activated immune cells in the periphery evoke these behavioral symptoms by driving inflammatory changes in the brain. However, experimental data in humans are lacking. Here we demonstrate in healthy male volunteers (10 endotoxin treated, 8 placebo treated) that intravenous administration of low-dose endotoxin (0.8 ng/kg body weight), a prototypical pathogen-associated molecular pattern that activates the innate immune system, not only induces a significant increase in peripheral blood cytokine concentrations (that is, tumor necrosis factor-α, interleukin (IL)-6, IL-10) but also results, with some latency, in a robust and selective increase of IL-6 in the cerebrospinal fluid (CSF). Moreover, we found a strong association between the endotoxin-induced increase of IL-6 in the CSF and the severity of mood impairment, with larger increases in CSF IL-6 concentration followed by a greater deterioration in mood. Taken together, these findings suggest that the appearance of depressive symptoms in inflammatory conditions might be primarily linked to an increase in central IL-6 concentration, identifying IL-6 as a potential therapeutic target in mood disorders.

What Makes You Feel Sick After Inflammation? Predictors of Acute and Persisting Physical Sickness Symptoms Induced by Experimental Endotoxemia.

We aimed to identify statistical predictor variables of lipopolysaccharide (LPS)-induced physical sickness symptoms during the acute and late inflammatory phases using multivariate regression analyses. Data from N = 128 healthy volunteers who received i.v. LPS injection (0.4 or 0.8 ng/kg) or placebo were pooled for analyses. Physical sickness symptoms experienced during the acute (0-6h postinjection) and late (6-24h postinjection) phases were assessed with the validated General-Assessment-of-Side-Effects (GASE) questionnaire. LPS-treated subjects reported significantly more physical sickness symptoms. Physical symptoms during the acute phase were associated with LPS-induced mood impairments and interleukin (IL)-6 increases, explaining 28.5% of variance in GASE scores. During late phase, LPS-induced increases in cortisol and IL-6 plasma concentrations and baseline depression were significant predictor variables, explaining 38.5% of variance. In patients with recurrent or chronic inflammatory states, these factors may act as risk factors ultimately contributing to an exacerbation of sickness symptoms, and should be considered as potential targets for therapeutic strategies.

Preventive effects of minocycline in a neurodevelopmental two-hit model with relevance to schizophrenia.

Maternal immune activation can increase the vulnerability of the offspring to develop neuroimmune and behavioral abnormalities in response to stress in puberty. In offspring of immune-challenged mothers, stress-induced inflammatory processes precede the adult onset of multiple behavioral dysfunctions. Here, we explored whether an early anti-inflammatory intervention during peripubertal stress exposure might prevent the subsequent emergence of adult behavioral pathology. We used an environmental two-hit model in mice, in which prenatal maternal administration of the viral mimetic poly(I:C) served as the first hit, and exposure to sub-chronic unpredictable stress during peripubertal maturation as the second hit. Using this model, we examined the effectiveness of the tetracycline antibiotic minocycline (MINO) given during stress exposure to block stress-induced inflammatory responses and to prevent subsequent behavioral abnormalities. We found that combined exposure to prenatal immune activation and peripubertal stress caused significant deficits in prepulse inhibition and increased sensitivity to the psychotomimetic drugs amphetamine and dizocilpine in adulthood. MINO treatment during stress exposure prevented the emergence of these behavioral dysfunctions. In addition, the pharmacological intervention blocked hippocampal and prefrontal microglia activation and interleukin-1ß expression in offspring exposed to prenatal infection and peripubertal stress. Together, these findings demonstrate that presymptomatic MINO treatment can prevent the subsequent emergence of multiple behavioral abnormalities relevant to human neuropsychiatric disorders with onset in early adulthood, including schizophrenia. Our epidemiologically informed two-hit model may thus encourage attempts to explore the use of anti-inflammatory agents in the early course of brain disorders that are characterized by signs of central nervous system inflammation during development.

Effects of Neurexan ® in an experimental acute stress setting--An explorative double-blind study in healthy volunteers.

AIMS: The objective of this study was to assess the efficacy profile of Nx4 (Neurexan ®) in an acute experimental stress setting. An acute stress reaction is a biopsychological condition arising in response to an event that is individually regarded as emotionally stressful. Medications can mitigate stress perception and stress reactions, but may also have side effects. MATERIALS AND METHODS: Sixty-four healthy male and female volunteers participated in this prospective two-arm two-site study following an explorative randomized placebo-controlled double-blind study design. Participants took six tablets of either Nx4 or placebo during a time period of 2.5h before exposure to an acute psychological stressor (Trier Social Stress Test), and were subsequently monitored for 1.5h. Subjective stress ratings as well as cardiovascular and neuroendocrine parameters were analyzed before and after stress exposure. KEY FINDINGS: All changes in primary and secondary efficacy parameters corresponded well with the experimental acute stress setting. Nx4 did not affect subjective stress ratings but significantly diminished stress-induced increases in salivary cortisol and plasma adrenaline. Nx4 was as safe as placebo and very well tolerated. SIGNIFICANCE: The results suggest an attenuated neuroendocrine stress response in healthy volunteers induced by Nx4. However, further investigations are needed to confirm these observations as well as to better understand why some parameters were affected while others were not. Future investigations should be extended to chronically stressed individuals with a greater disposition to experience stress in everyday life. ClinicalTrials.gov Identifier: NCT01703819.

Extinction of conditioned taste aversion is related to the aversion strength and associated with c-fos expression in the insular cortex.

Taste aversion learning is a type of conditioning where animals learn to associate a novel taste (conditioned stimulus; CS) with a stimulus inducing symptoms of poisoning or illness (unconditioned stimulus; US). As a consequence animals later avoid this taste, a reaction known as conditioned taste aversion (CTA). An established CTA extinguishes over time when the CS is repeatedly presented in the absence of the US. However, inter-individual differences in CTA extinction do exist. Using a model of behavioral conditioning with saccharin as CS and the immunosuppressant cyclosporine A as US, the present study aimed at further elucidating the factors underlying individual differences in extinction learning by investigating whether extinction of an established CTA is related to the strength of the initially acquired CS-US association. In addition, we analyzed the expression of the neuronal activation marker c-fos in brain structures relevant for acquisition and retrieval of the CTA, such as the insular cortex and the amygdala. We here show that animals, displaying a strong CS-US association during acquisition, maintained a strong CTA during unreinforced CS re-exposures, in contrast to animals with moderate CS-US association. Moreover, the latter animals showed increased c-fos mRNA expression in the insular cortex. Our data indicate that CTA extinction apparently depends on the strength of the initially learned CS-US association. In addition, these findings provide further evidence that the memory for the initial excitatory conditioning and its subsequent extinction is probably stored in those structures that participate in the processing of the CS and the US.

Are there sex differences in placebo analgesia during visceral pain processing? A fMRI study in healthy subjects.

BACKGROUND: We explored sex differences in the neural mechanisms mediating placebo analgesia in an established visceral pain model involving painful rectal distensions in healthy volunteers. METHODS: N = 15 men and N = 15 women underwent three consecutive functional magnetic resonance imaging sessions during which cued painful rectal distensions were delivered. After an adaptation session, positive expectations were induced with deceptive instructions regarding administration of an analgesic drug (placebo session). In the other session (control), truthful information about an inert substance was given. Sex differences in placebo-induced modulation of neural activation during anticipation and pain were analyzed along with ratings of expected and perceived pain intensity. KEY RESULTS: Placebo-induced reductions in pain ratings were comparable between men and women. At the level of the brain, group comparisons with respect to differences between the placebo and control conditions revealed greater modulation of the posterior insula (regions-of-interest analysis: pFWE < 0.05) and dorsolateral prefrontal cortex (whole-brain analysis: p < 0.001, uncorrected) during pain anticipation in women. During pain, placebo-induced down-regulation of the insula was altered in women compared to men (ROI analysis: pFWE < 0.05). CONCLUSIONS & INFERENCES: Our data provide first evidence supporting sex differences in pain-induced neural modulation during visceral placebo analgesia despite similar placebo-induced reductions in perceived pain intensity. These preliminary findings might contribute to elucidating mechanisms mediating placebo effects in clinical conditions associated with chronic abdominal pain such as in irritable bowel syndrome.

Preserving learned immunosuppressive placebo response: perspectives for clinical application.

Akin to other physiological responses, immune functions can be modified through behavioral conditioning as part of a learned placebo response. However, like every learning process, learned immune responses are subject to extinction. We analyzed the extinction of learned immunosuppression in healthy male volunteers, using an established conditioning paradigm with the immunosuppressive drug cyclosporin A (CsA) as unconditioned stimulus (US) and a gustatory stimulus as conditioned stimulus (CS). We observed a learned suppression of T-cell function after two and four reexposures to the CS, which was extinguished after 14 unreinforced CS reexposures. However, administration of "subtherapeutic" CsA dosages together with the CS counteracted the extinction of the learned immunosuppression. These findings provide the basis for a potentially successful implementation of conditioning paradigms as supportive therapy to immunopharmacological regimens in clinical settings. The aim is to reduce the required amount of medication while maximizing the therapeutic outcome for the patient's benefit.

Perceived treatment group affects behavioral and neural responses to visceral pain in a deceptive placebo study.

BACKGROUND: To assess effects of perceived treatment (i.e. drug vs placebo) on behavioral and neural responses to rectal pain stimuli delivered in a deceptive placebo condition. METHODS: This fMRI study analyzed the behavioral and neural responses during expectation-mediated placebo analgesia in a rectal pain model. In N = 36 healthy subjects, the blood oxygen level-dependent (BOLD) response during cued anticipation and painful stimulation was measured after participants were informed that they had a 50% chance of receiving either a potent analgesic drug or an inert substance (i.e., double-blind administration). In reality, all received placebo. We compared responses in subjects who retrospectively indicated that they received the drug and those who believed to have received placebo. KEY RESULTS: 55.6% (N = 20) of subjects believed that they had received a placebo, whereas 36.1% (N = 13) believed that they had received a potent analgesic drug. Subjects who were uncertain (8.3%, N = 3) were excluded. Rectal pain-induced discomfort was significantly lower in the perceived drug treatment group (P < 0.05), along with significantly reduced activation of the insular, the posterior and anterior cingulate cortices during pain anticipation, and of the anterior cingulate cortex during pain (all P < 0.05 in regions-of-interest analyses). CONCLUSIONS & INFERENCES: Perceived treatment constitutes an important aspect in placebo analgesia. A more refined understanding of individual treatment expectations and perceived treatment allocation has multiple implications for the design and interpretation of clinical trials and experimental studies on placebo and nocebo effects.

How positive and negative expectations shape the experience of visceral pain: an experimental pilot study in healthy women.

BACKGROUND: In order to elucidate placebo and nocebo effects in visceral pain, we analyzed the effects of positive and negative expectations on rectal pain perception, rectal pain thresholds, state anxiety and cortisol responses in healthy women. METHODS: Painful rectal distensions were delivered at baseline, following application of an inert substance combined with either positive instructions of pain relief (placebo group, N = 15), negative instructions of pain increase (nocebo group, N = 17), or neutral instructions (control, N = 15). Perceived pain intensity, unpleasantness/aversion and urge-to-defecate, state anxiety and serum cortisol were determined at baseline, immediately following group-specific instructions and on a second study day after the same instructions (test day). Rectal pain thresholds were determined at baseline and on the test day. KEY RESULTS: Whereas perceived pain intensity was significantly decreased in the placebo group, the nocebo group revealed significantly increased pain intensity ratings, along with significantly greater anticipatory anxiety on the test day (all P < 0.05 vs controls). Cortisol concentrations were significantly increased in the nocebo group following treatment but not on the test day. CONCLUSIONS & INFERENCES: The experience of abdominal pain can be experimentally increased or decreased by inducing positive or negative expectations. Nocebo effects involve a psychological stress response, characterized by increased anticipatory anxiety. These findings further underscore the role of cognitive and emotional factors in the experience of visceral pain, which has implications for the pathophysiology and treatment of patients with chronic abdominal complaints.

Behavioural and neural correlates of visceral pain sensitivity in healthy men and women: does sex matter?

INTRODUCTION: We assessed sex differences in behavioural and neural responses to rectal pain stimuli in healthy subjects. METHODS: In age- and body mass index-matched healthy subjects (n = 15 men, 15 women), rectal sensory and pain thresholds were assessed with a pressure-controlled barostat device. The blood oxygen level-dependent response during cued anticipation and painful stimulation was measured using functional magnetic resonance imaging (fMRI). Retrospective pain evaluations were accomplished with visual analogue scales. For fMRI data, region-of-interest (ROI) analyses and additional whole-brain analyses were carried out. RESULTS: There were no sex differences in rectal thresholds or pain ratings. ROI analyses revealed comparable distension-induced activation of the thalamus, somatosensory cortex, insula and dorsolateral prefrontal cortex (DLPFC). Only in additional whole-brain analyses did we find increased activation in women in DLPFC and middle temporal gyrus during pain anticipation and in the cerebellum and medial frontal gyrus during pain. A significant inverse association between rectal pain threshold and distension-induced activation in virtually all ROIs was found in women. In men, pain thresholds and insula activation were positively correlated, as were pain ratings and anterior cingulate cortex activation. CONCLUSIONS: Healthy men and women do not differ in behavioural measures of visceral pain sensitivity. The pattern of neural activation is comparable in the majority of pain-processing brain regions, although women may differ in the activation of DLPFC which could reflect sex differences in cognitive-emotional pain regulation. Women with lower pain thresholds showed greater neural responses, which may be relevant in the pathophysiology of visceral hyperalgesia.

Plasma noradrenaline and state anxiety levels predict placebo response in learned immunosuppression.

Large interindividual differences exist in the presence and extent of placebo responses in both experimental and clinical studies, but little is known about possible predictors of these responses. We employed a behaviorally conditioned immunosuppression paradigm in healthy men to analyze predictors of learned placebo responses. During acquisition, the subjects received either the immunosuppressant cyclosporin A (n = 32) or a placebo (n = 14) (unconditioned stimuli (US)) together with a novel-tasting drink (conditioned stimulus (CS)). During evocation, the subjects were reexposed to the CS alone. In responders (n = 15), the CS alone caused a significant inhibition of interleukin (IL)-2 production by anti-CD3-stimulated peripheral blood T cells, closely mimicking the drug effect. Nonresponders (n = 17) did not show responses different from those of the controls. Multiple-regression analyses showed that baseline IL-2, plasma noradrenaline, and state anxiety predicted nearly 60% of the variance in the conditioned IL-2 response. These data provide first evidence for putative biological and psychological predictors of learned placebo responses.

Subclinical depressive symptoms affect responses to acute psychosocial stress in healthy premenopausal women.

Subclinical depressive symptoms constitute a primary risk factor for major depression as well as for cardiovascular conditions, which may be mediated by endocrine or immune alterations. The aim of this study was to assess the association between the extent of subclinical depressive symptoms and neuroendocrine and immune cell responses to acute psychosocial stress in healthy females. In N = 33 healthy premenopausal women, state anxiety, plasma adrenocorticotropic hormone and serum cortisol, and interleukin-6 (IL-6) concentration responses to public speaking stress were assessed. Beck depression inventory (BDI) scores were entered as a covariate in the analyses. The IL-6 response was significantly associated with BDI scores (p < 0.05). Secondary analyses revealed that women with more subclinical depressive symptoms demonstrated a reduced stress-induced increase in circulating IL-6 level (p < 0.05). By contrast, stress-induced neuroendocrine activation was not associated with depressive symptoms. Hence, subclinical depressive symptoms were associated with IL-6 responses to stress in young, healthy women. Unexpectedly, there was a reduced increase of serum IL-6 level in response to stress. Effects of depressive symptoms on the IL-6 response to stress may differ between subclinical and major depression.

Affective disturbances modulate the neural processing of visceral pain stimuli in irritable bowel syndrome: an fMRI study.

OBJECTIVE: To address the role of anxiety and depression symptoms in altered pain processing in irritable bowel syndrome (IBS). DESIGN: In this functional magnetic resonance imaging study, the blood oxygen level-dependent (BOLD) response to rectal distensions delivered at previously determined individual discomfort thresholds was assessed. PATIENTS: 15 female patients with irritable bowel syndrome (IBS) and with normal rectal pain thresholds, and 12 healthy women. MEASURES: The correlation of anxiety and depression symptoms, measured with the Hospital Anxiety and Depression Scale (HADS), with subjective pain ratings and the BOLD response during distension-induced brain activation were analysed within IBS. Group differences in pain-induced brain activation with and without controlling for HADS scores were evaluated. RESULTS: Patients with IBS experienced significantly more pain and discomfort upon rectal distensions in the scanner, despite unaltered rectal sensory thresholds. Anxiety and depression scores were associated with these subjective stimulus ratings, but not with rectal sensory thresholds. Anxiety symptoms in IBS were significantly associated with pain-induced activation of the anterior midcingulate cortex and pregenual anterior cingulate cortex. Depression scores correlated with activation of the prefrontal cortex (PFC) and cerebellar areas within IBS. Group comparisons with the two-sample t test revealed significant activation in the IBS versus controls contrast in the anterior insular cortex and PFC. Inclusion of anxiety and depression scores, respectively, as confounding variables led to a loss of significant group differences. CONCLUSIONS: Altered central processing of visceral stimuli in IBS is at least in part mediated by symptoms of anxiety and depression, which may modulate the affective-motivational aspects of the pain response.

Effects of psychological stress on the cerebral processing of visceral stimuli in healthy women.

The aim of the study was to analyse effects of psychological stress on the neural processing of visceral stimuli in healthy women. The brain functional magnetic resonance imaging blood oxygen level-dependent response to non-painful and painful rectal distensions was recorded from 14 healthy women during acute psychological stress and a control condition. Acute stress was induced with a modified public speaking stress paradigm. State anxiety was assessed with the State-Trait-Anxiety Inventory; chronic stress was measured with the Perceived Stress Questionnaire. During non-painful distensions, activation was observed in the right posterior insular cortex (IC) and right S1. Painful stimuli revealed activation of the bilateral anterior IC, right S1, and right pregenual anterior cingulate cortex. Chronic stress score was correlated with activation of the bilateral amygdala, right posterior IC (post-IC), left periaqueductal grey (PAG), and right dorsal posterior cingulate gyrus (dPCC) during non-painful stimulation, and with activation of the right post-IC, right PAG, left thalamus (THA), and right dPCC during painful distensions. During acute stress, state anxiety was significantly higher and the acute stress - control contrast revealed activation of the right dPCC, left THA and right S1 during painful stimulation. This is the first study to demonstrate effects of acute stress on cerebral activation patterns during visceral pain in healthy women. Together with our finding that chronic stress was correlated wit the neural response to visceral stimuli, these results provide a framework for further studies addressing the role of chronic stress and emotional disturbances in the pathophysiology of visceral hyperalgesia.

Prevalence and implications of anxiety in polycystic ovary syndrome: results of an internet-based survey in Germany.

BACKGROUND: Comparatively little attention has been paid to the symptoms of anxiety in polycystic ovary syndrome (PCOS), although anxiety disorders constitute the most common psychiatric diagnoses among endocrine patients and in the general population. Therefore, our goal was to address the prevalence, determinants and implications of anxiety alone or anxiety in combination with depression in German women with PCOS. METHODS: In this nation-wide, internet-based survey, anxiety and depression (Hospital Anxiety and Depression Scale, HADS) and quality of life (SF-12) were assessed together with sociodemographic information and clinical PCOS symptoms in 448 PCOS women. RESULTS: Of the patients, 34% showed clinically relevant HADS anxiety scores and 21% had clinically relevant HADS depression scores. Quality of life was significantly impaired in PCOS women with anxiety (P < 0.001), in particular, in women with comorbid anxiety and depression (P < 0.001). The risk for clinically relevant HADS anxiety scores was significantly enhanced in PCOS women with acne (odds ratio (OR) = 1.52; 95% confidence interval (CI) = 1.03-2.52) and an unfulfilled wish to conceive (OR = 1.50; 95% CI = 1.01-2.23). CONCLUSIONS: PCOS women may be at an increased risk for clinically relevant anxiety, and comorbid anxiety and depression is also very common. Anxiety contributes to impaired quality of life in PCOS. Given the high prevalence and the serious implications, and the availability of effective treatment options given proper diagnosis, clinicians should be more aware of anxiety disorders in women with PCOS.