Mitochondrial proteolytic stress induced by loss of mortalin function is rescued by Parkin and PINK1.

The mitochondrial chaperone mortalin was implicated in Parkinson's disease (PD) because of its reduced levels in the brains of PD patients and disease-associated rare genetic variants that failed to rescue impaired mitochondrial integrity in cellular knockdown models. To uncover the molecular mechanisms underlying mortalin-related neurodegeneration, we dissected the cellular surveillance mechanisms related to mitochondrial quality control, defined the effects of reduced mortalin function at the molecular and cellular levels and investigated the functional interaction of mortalin with Parkin and PINK1, two PD-related proteins involved in mitochondrial homeostasis. We found that reduced mortalin function leads to: (1) activation of the mitochondrial unfolded protein response (UPR(mt)), (2) increased susceptibility towards intramitochondrial proteolytic stress, (3) increased autophagic degradation of fragmented mitochondria and (4) reduced mitochondrial mass in human cells in vitro and ex vivo. These alterations caused increased vulnerability toward apoptotic cell death. Proteotoxic perturbations induced by either partial loss of mortalin or chemical induction were rescued by complementation with native mortalin, but not disease-associated mortalin variants, and were independent of the integrity of autophagic pathways. However, Parkin and PINK1 rescued loss of mortalin phenotypes via increased lysosomal-mediated mitochondrial clearance and required intact autophagic machinery. Our results on loss of mortalin function reveal a direct link between impaired mitochondrial proteostasis, UPR(mt) and PD and show that effective removal of dysfunctional mitochondria via either genetic (PINK1 and Parkin overexpression) or pharmacological intervention (rapamycin) may compensate mitochondrial phenotypes.

Hyperventilation and attention: effects of hypocapnia on performance in a stroop task.

This study aimed to investigate the effect of hypocapnia on attentional performance. Hyperventilation, producing hypocapnia, is associated with physiological changes in the brain and with subjective symptoms of dizziness, concentration problems and derealization. In this study (N=42), we examined cognitive performance on a Stroop-like task, following either 3 min of hypocapnic or normocapnic overbreathing. Both overbreathing trials were run on separate days, each preceded by a baseline trial with the same task during normal breathing. More and other symptoms were reported after hypocapnia compared to normocapnia. Also, more errors were made and progressively slower reaction times (RT's) were observed during recovery from hypocapnia. These performance deficits were only found in participants characterized by apneas. The number of symptoms did not correlate with RT's or errors. The pattern of data suggested that hypoxia, as a result of apneas during recovery from hypocapnia, caused the cognitive performance deficit.

Psychosomatic symptoms and breathing pattern.

OBJECTIVE: Study of the links between breathing pattern, negative affectivity, and psychosomatic complaints at rest and following hyperventilation. METHODS: In 819 patients with anxiety and somatoform disorders and 159 healthy subjects, self-reported symptoms, breathing pattern, and end-tidal CO(2) concentration (FetCO(2)) were recorded during rest and following a hyperventilation provocation test (HVPT). The relationship between disorder category, symptoms, age, and score of STAI-trait (as a measure of negative affectivity) on the one hand, and breathing pattern on the other was investigated, separately in men and women. RESULTS: Anxiety disorders, and to a lesser extent, somatoform disorders, were characterized by breathing instability (progressive decrease of FetCO(2) at rest during mouthpiece breathing, delayed recovery of FetCO(2) following HVPT), the mean values of respiratory frequency, and FetCO(2) being modulated by STAI-trait. After grouping the symptoms into independent factors, links were observed between symptoms and breathing pattern, independently from the presence of an anxiety or somatoform disorder. CONCLUSION: Some symptom factors appeared to be related to a lower FetCO(2) during hyperventilation, others likely directly influenced the breathing pattern. Among those, mainly respiratory symptoms were accompanied by a reduction of FetCO(2) at rest, with slower recovery of FetCO(2) following HVPT. The latter was observed also in the presence of marked anxiety. In contrast, subjects complaining of dizziness, fainting, and paresthesias in daily life presented higher values of FetCO(2) following HVPT, probably due to a voluntary braking of ventilation during HVPT.

Generalization of acquired somatic symptoms in response to odors: a pavlovian perspective on multiple chemical sensitivity.

OBJECTIVE: Somatic symptoms that occur in response to odors can be acquired in a pavlovian conditioning paradigm. The present study investigated 1) whether learned symptoms can generalize to new odors, 2) whether the generalization gradient is linked to the affective or irritant quality of the new odors, and 3) whether the delay between acquisition and testing modulates generalization. METHODS: Conditional odor stimuli (CS) were (diluted) ammonia and niaouli. One odor was mixed with 7.4% CO2-enriched air (unconditional stimulus) during 2-minute breathing trials (CS+ trial), and the other odor was presented with air (CS- trial). Three CS+ and three CS- trials were conducted in a semirandomized order (acquisition phase). The test phase involved one CS+-only (CS+ without CO2) and one CS- test trial, followed by three trials using new odors (butyric acid, acetic acid, and citric aroma). Half of the subjects (N = 28) were tested immediately, and the other half were tested after 1 week. Ventilatory responses were measured during and somatic symptoms were measured after each trial. RESULTS: Participants had more symptoms in response to CS+-only exposures, but only when ammonia was used as the CS+. Also, generalization occurred: More symptoms were reported in response to butyric and acetic acid than to citric aroma and only in participants who had been conditioned. Both the selective conditioning and the generalization effect were mediated by negative affectivity of the participants. The delay between the acquisition and test phases had no effect. CONCLUSIONS: Symptoms that occur in response to odorous substances can be learned and generalize to new substances, especially in persons with high negative affectivity. The findings further support the plausibility of a pavlovian perspective of multiple chemical sensitivity.

Acquisition and extinction of somatic symptoms in response to odours: a Pavlovian paradigm relevant to multiple chemical sensitivity.

OBJECTIVES: Multiple chemical sensitivity is a poorly understood syndrome in which various symptoms are triggered by chemically unrelated, but often odorous substances, at doses below those known to be harmful. This study focuses on the process of pavlovian acquisition and extinction of somatic symptoms triggered by odours. METHODS: Diluted ammonia and butyric acid were odorous conditioned stimuli (CS). The unconditioned stimulus (US) was 7.4% CO2 enriched air. One odour (CS+) was presented together with the US for 2 minutes (CS+ trial), and the other odour (CS-) was presented with air (CS-trial). Three CS+ and three CS-exposures were run in a semi-randomised order; this as the acquisition (conditioning) phase. To test the effect of the conditioning, each subject then had one CS+ only--that is, CS+ without CO2--and one CS- test exposure. Next, half the subjects (n = 32) received five additional CS+ only exposures (extinction group), while the other half received five exposures to breathing air (wait group). Finally, all subjects got one CS+ only test exposure to test the effect of the extinction. Ventilatory responses were measured during and somatic symptoms after each exposure. RESULTS: More symptoms were reported upon exposure to CS+ only than to CS-odours, regardless of the odour type. Altered respiratory rate was only found when ammonia was CS+. Five extinction trials were sufficient to reduce the level of acquired symptoms. CONCLUSION: Subjects can acquire somatic symptoms and altered respiratory behaviour in response to harmless, but odorous chemical substances, if these odours have been associated with a physiological challenge that originally had caused these symptoms. The conditioned symptoms can subsequently be reduced in an extinction procedure. The study further supports the plausibility of a pavlovian conditioning hypothesis to explain the pathogenesis of MCS.

Fear-relevant images as conditioned stimuli for somatic complaints, respiratory behavior, and reduced end-tidal pCO2.

Two fear-relevant imagery scripts were used as conditioned stimuli (CSs) in a differential learning paradigm with 5.5% CO2-enriched air as unconditioned stimulus (US). In another condition, 2 neutral scripts were used as CSs (N = 56). Within each condition, one script was imagined while breathing the CO2-enriched air (CS+/US trial), the other while breathing room air (CS- trial). Three CS+ and 3 CS- trials were run in an acquisition phase, followed by 2 CS+ and 2 CS- test trials (imagining the scripts while breathing air). Respiratory behavior, end-tidal CO2, and heart rate were measured throughout the experiment; subjective symptoms were measured after each trial. The type of imagery had strong effects on symptoms and physiological responses. A selective conditioning effect was also observed: CS+ imagery produced more symptoms and altered respiratory behavior compared with CS- imagery, but only in the fear-relevant script condition. The findings are discussed as to their relevance for panic and agoraphobic anxiety.

Subjective symptoms and breathing pattern at rest and following hyperventilation in anxiety and somatoform disorders.

The purpose of the present study was to investigate the diagnostic specificity of bodily symptoms and respiratory behavior at rest and after a hyperventilation provocation test (HVPT) in patients that were either grouped according to the DSM classification or diagnosed as suffering from hyperventilation syndrome. Nine hundred three anxiety and somatoform patients, showing symptoms supposedly caused by psychogenic hyperventilation, and 170 healthy subjects, were studied. Breathing pattern and end-tidal CO2 concentration were recorded during breathing at rest and following a HVPT. Subjective symptoms in daily life and after HVPT were measured. A principal-components analysis was performed on both the symptoms and breathing variables and their specificity levels were compared in the two classifications of patients. Some symptoms in daily life were grouped together with the same symptoms after the HVPT, other symptoms were not. This suggests that the HVPT elicited partly specific symptoms, and partly reproduced the symptoms experienced in daily life. Similar findings were observed with respect to the breathing variables. Patients with panic differed from other patients with anxiety disorders by an increased level of symptoms and a FETCO2 decline at rest. The HVPT may be informative for diagnosis because it provokes some of the typical somatic and psychological symptoms, and it identifies the breathing instability that is characteristic of both patients with HVS and with anxiety. The same symptoms and breathing variables characterized the patients, whatever their classification. Overall, the specificity of breathing variables is rather low.

Negative affect, respiratory reactivity, and somatic complaints in a CO2 enriched air inhalation paradigm.

Subjects scoring high on negative affectivity (NA) are known to report more psychosomatic complaints than subjects scoring low. According to the symptom perception hypothesis, high NA subjects attend more to somatic sensations and interpret these as more threatening. We investigated the relationship between NA and psychosomatic complaints in a group of high and low NA subjects (N = 72) in, (a) a questionnaire study, and (b) in a laboratory setting. The latter involved the inhalation of three different gas mixtures (room air, 5.5% and 7.5% CO2-enriched air) while respiratory responses were registered. Subjective complaints were measured after each trial. High NA subjects reported more complaints than low NA subjects in the questionnaire study. However, NA had no main effects on complaints in the laboratory study and did not interact with the effects of gas mixture on complaints. During room air trials, NA correlated only with general arousal complaints when a strong respiratory challenge had not been given before. The pattern of results suggests that experimental inductions of complaints may largely wipe out NA-related differences in attentional/interpretative processes that may mediate the NA complaints link.

Memory effects on symptom reporting in a respiratory learning paradigm.

With odors as conditioned stimuli (CSs) and CO2-enriched air as the unconditioned stimulus, participants learned to exhibit respiratory responses and somatic complaints on presentation of only the odor CS+. Studied was whether complaints during CS+-only trials were inferred from the conditioned somatic responses or were based on activated memory of the complaints during acquisition. Participants (N = 56) were either attentionally directed away or not from the complaints during acquisition, and the effects on somatic complaints during test were studied. Respiratory responses, heart rate, and somatic complaints were measured. No physiological conditioning effects were found. However, more complaints were reported to the CS+ than to the CS- odor, but only when the CS+ was foul smelling. This effect was modulated by the attention manipulation, showing that the learned complaints during the test phase were based on memory of the acquisition complaints and not on physiological responses during the test.

Influence of awareness of the recording of breathing on respiratory pattern in healthy humans.

This study was designed to test whether awareness of the measurement of breathing influences the breathing pattern in healthy subjects under routine laboratory conditions. Seventy four subjects (40 females and 34 males), aged 21-63 yrs, were studied under three different conditions whilst their breathing was being recorded for 5 min by means of inductance plethysmography (Respitrace): 1) subjects were misled into believing that their breathing was not being recorded but that they had to wait for 5 min whilst equipment was calibrated; 2) subjects were instructed that their breathing pattern was being recorded for 5 min; 3) the subject's breathing was recorded for 5 min with mouthpiece and pneumotachograph. The first two conditions were randomized. The Respitrace was calibrated by means of multiple linear regression carried out during the 5 min period of quiet breathing through a mouthpiece. Awareness of the recording of breathing caused prolongation of inspiratory (tI) and expiratory time (tE). Breathing through the mouthpiece resulted in an increase of tI, tE and tidal volume (VT). The breathing irregularities (sighs and end-expiratory pauses) decreased when subjects were aware of the recording of breathing and nearly disappeared when subjects breathed through the mouthpiece. The end-tidal carbon dioxide concentration was not significantly different between the three conditions. Mouthpiece breathing often induced some respiratory discomfort and even anxiety, particularly in females. Awareness by the subject that his or her breathing was being recorded altered the spontaneous breathing pattern, mainly the breathing frequency. In addition, use of a mouthpiece markedly increased tidal volume, particularly in females in whom mouthpiece breathing induced more complaints than in males.

Unsteadiness of breathing in patients with hyperventilation syndrome and anxiety disorders.

The breathing pattern of 399 patients with hyperventilation syndrome (HVS) and/or with anxiety disorders and that of 347 normal controls was investigated during a 5 min period of quiet breathing and after a 3 min period of voluntary hyperventilation. The diagnosis of HVS was based on the presence of several suggestive complaints occurring in the context of stress, and reproduced by voluntary hyperventilation. Organic diseases as a cause of the symptoms were excluded. The anxiety disorders were diagnosed by means of an abbreviated version of the Anxiety Disorders Interview Schedule (ADIS). There was a large overlap between the two diagnoses. Simply breathing via a mouthpiece and pneumotachograph made end-tidal CO2 fractional concentration (FET,CO2) decrease progressively both in hyperventilators and in patients with anxiety disorders, but not in normals. At the start of the measurement the FET,CO2 was not different between patients and healthy subjects. In patients < or = 28 yrs, the decrease of FET,CO2 resulted from a higher tidal volume, and in patients > or = 29 years from an increase in frequency. After voluntary hyperventilation, the recovery of FET,CO2, was delayed in patients, due to a slower normalization of respiratory frequency in females and in older males, and of tidal volume in younger males, and also due to less frequent end-expiratory pauses. When breathing was recorded first by means of inductive plethysmography (Respitrace), the progressive decline of FET,CO2 seen in patients was not observed: from the onset of the recording, FET,CO2 was reduced in patients. It did not change further when, immediately afterwards, the subject switched to mouthpiece breathing. The finding that breathing through a mouthpiece induces hyperventilation in patients and that recovery of FET,CO2 is delayed after voluntary hyperventilation, suggests that the respiratory control system is less resistant to challenges (mouthpiece or voluntary hyperventilation) in those patients. On the other hand, the lower values of FET,CO2 measured during recording by means of a Respitrace probably result from a challenge, prior to the recordings, induced by the fitting of the measuring device to the patient. This unsteadiness of breathing characterizes patients with hyperventilation syndrome and those with anxiety disorders, but is not sufficiently sensitive to be used for individual diagnosis.

Learning to have psychosomatic complaints: conditioning of respiratory behavior and somatic complaints in psychosomatic patients.

OBJECTIVE: Assuming a subjective similarity between the experience of a hyperventilation episode and inhaling CO2-enriched air, we tested whether a respiratory challenge in association with a particular stimulus could result in altered respiratory behavior and associated somatic complaints upon presenting the stimulus only. METHOD: Psychosomatic patients (N = 28) reporting hyperventilation complaints participated in a differential conditioning paradigm using odors with a positive or negative valence as conditioned stimuli (CS+ or CS-) and 7.4% CO2-enriched air as the unconditioned stimulus (US). Three CS+ and three CS-acquisition trials were run. During the test phase, two CS(+)- and two CS(-)-only trials were run, followed by two new test odors (with a positive or negative valence). Respiratory frequency, tidal volume, end-tidal fractional concentration of CO2, and heart rate were measured throughout the experiment. Somatic complaints were registered after each trial. RESULTS: We observed a) increased respiratory frequency and an elevated level of somatic complaints upon presenting the CS+ only; b) a selective association effect: conditioning was only apparent with the negatively valenced CS+ odor; (c) no generalization of respiratory responses and complaints to the new odors; (d) no conditioning effect on dummy complaints that are usually not reported when inhaling CO2; (e) in exploratory comparisons with normal subjects, stronger conditioning effects on typical hyperventilation complaints in patients, and, in female subjects, on respiratory frequency. CONCLUSION: Respiratory responses and psychosomatic complaints can be elicited by conditioned stimuli in a highly specific way. The findings are relevant for disorders in which respiratory abnormalities and/or psychosomatic complaints may play a role and for multiple chemical sensitivity.

Influence of breathing therapy on complaints, anxiety and breathing pattern in patients with hyperventilation syndrome and anxiety disorders.

The effect of breathing therapy was evaluated in patients with hyperventilation syndrome (HVS). The diagnosis of HVS was based on the presence of several suggestive complaints occurring in the context of stress, and reproduced by voluntary hyperventilation. Organic diseases as a cause of the symptoms were excluded. Most of these patients met the criteria for an anxiety disorder. The therapy was conducted in the following sequence: (1) brief, voluntary hyperventilation to reproduce the complaints in daily life: (2) reattribution of the cause of the symptoms to hyperventilation: (3) explaining the rationale of therapy-reduction of hyperventilation by acquiring an abdominal breathing pattern, with slowing down of expiration: and (4) breathing retraining for 2 to 3 months by a physiotherapist. After breathing therapy, the sum scores of the Nijmegen Questionnaire were markedly reduced. Improvements were registered in 10 of the 16 complaints of the questionnaire. The level of anxiety evaluated by means of the State-Trait Anxiety Inventory (STAI) decreased slightly. The breathing pattern was modified significantly after breathing retraining. Mean values of inspiration and expiration time and tidal volume increased, but end-tidal CO2 concentration (FETCO2) was not significantly modified except in the group of younger women (< or = 28 years). A canonical correlation analysis relating the changes of the various complaints to the modifications of breathing variables showed that the improvement of the complaints was correlated mainly with the slowing down of breathing frequency. The favorable influence of breathing retraining on complaints thus appeared to be a consequence of its influence primarily on breathing frequency, rather than on FETCO2.