Transgene-free, virus-based gene silencing in plants by artificial microRNAs derived from minimal precursors.

Artificial microRNAs (amiRNAs) are highly specific, 21-nucleotide (nt) small RNAs designed to silence target transcripts. In plants, their application as biotechnological tools for functional genomics or crop improvement is limited by the need of transgenically expressing long primary miRNA (pri-miRNA) precursors to produce the amiRNAs in vivo. Here, we analyzed the minimal structural and sequence requirements for producing effective amiRNAs from the widely used, 521-nt long AtMIR390a pri-miRNA from Arabidopsis thaliana. We functionally screened in Nicotiana benthamiana a large collection of constructs transiently expressing amiRNAs against endogenous genes and from artificially shortened MIR390-based precursors and concluded that highly effective and accurately processed amiRNAs can be produced from a chimeric precursor of only 89 nt. This minimal precursor was further validated in A. thaliana transgenic plants expressing amiRNAs against endogenous genes. Remarkably, minimal but not full-length precursors produce authentic amiRNAs and induce widespread gene silencing in N. benthamiana when expressed from an RNA virus, which can be applied into leaves by spraying infectious crude extracts. Our results reveal that the length of amiRNA precursors can be shortened without affecting silencing efficacy, and that viral vectors including minimal amiRNA precursors can be applied in a transgene-free manner to induce whole-plant gene silencing.

Predictability of the community-function landscape in wine yeast ecosystems.

Predictively linking taxonomic composition and quantitative ecosystem functions is a major aspiration in microbial ecology, which must be resolved if we wish to engineer microbial consortia. Here, we have addressed this open question for an ecological function of major biotechnological relevance: alcoholic fermentation in wine yeast communities. By exhaustively phenotyping an extensive collection of naturally occurring wine yeast strains, we find that most ecologically and industrially relevant traits exhibit phylogenetic signal, allowing functional traits in wine yeast communities to be predicted from taxonomy. Furthermore, we demonstrate that the quantitative contributions of individual wine yeast strains to the function of complex communities followed simple quantitative rules. These regularities can be integrated to quantitatively predict the function of newly assembled consortia. Besides addressing theoretical questions in functional ecology, our results and methodologies can provide a blueprint for rationally managing microbial processes of biotechnological relevance.

Allergic contact dermatitis to woods of the genus Dalbergia in musical instruments.

BACKGROUND: The woods of trees of the genus Dalbergia are a frequent cause of allergic contact dermatitis. OBJECTIVES: To describe the different patterns of symptoms and sensitisation to woods of the genus Dalbergia in four patients whose regular work is linked to music, either as performers (two patients) or instrument makers. RESULTS: The patients showed extensive cross-reactivity between the different Dalbergia woods. The two musicians exhibited symptoms exclusively in the contact areas, while the instrument makers showed a pattern of airborne symptoms. The period from sensitisation to the development of symptoms varied greatly among the patients. CONCLUSIONS: The symptoms of allergic contact dermatitis to Dalbergia woods differ markedly depending on the route of contact. Cross-reactivity between the woods of this genus is very wide. The sensitising potential varies between the various Dalbergia woods, which require different contact periods depending on their sensitising power.

Comparative Analysis of Psychophysiological Responses in Fibromyalgia Patients: Evaluating Neuromodulation Alone, Neuromodulation Combined with Virtual Reality, and Exercise Interventions.

BACKGROUND AND OBJECTIVES: Fibromyalgia, a chronic condition, manifests as widespread musculoskeletal pain, fatigue, sleep disturbances, autonomic and cognitive dysfunction, hypersensitivity to stimuli, and various somatic and psychiatric symptoms. This study, a controlled and randomized experiment, aimed to evaluate and compare the immediate effects of different treatments on fibromyalgia patients. MATERIALS AND METHODS: The treatments included the EXOPULSE Mollii suit, a combination of the EXOPULSE Mollii suit with a virtual reality (VR) protocol, and a physical exercise regimen. A cohort of 89 female fibromyalgia patients was randomly assigned to one of four groups: Control (n = 20), Suit only (n = 22), Suit combined with VR (n = 21), and Exercise (n = 26). RESULTS: This study found notable differences across the groups in several key parameters. In the Control group, significant changes were observed in Forced Expiratory Volume (FEV 1/FEV 6), the Numeric Rating Scale (NRS) for pain, Pressure Pain Threshold (PPT) at the epicondyle, cortical arousal levels, the 10 m up-and-go test, and in all measured variables related to temperature and muscle oxygenation. For the group using the suit alone, there were significant differences noted in the NRS, the chair stand test, palm temperature, and all muscle oxygenation parameters. The Suit + VR group showed significant changes in the NRS, PPT at the knee, handgrip strength test, the 10 m up-and-go test, one-leg balance test with the right leg, muscle oxygen saturation (SmO2), deoxygenated hemoglobin (HHb), and oxygenated hemoglobin (O2Hb). Finally, the Exercise group exhibited significant differences in FEV 1/FEV 6, chest perimeter difference, NRS, PPT at both the epicondyle and knee, cortical arousal, the chair stand test, the 10-m up-and-go test, and in SmO2, HHb, and O2Hb levels. CONCLUSIONS: combining neuromodulation with VR and targeted exercise regimens can effectively alleviate fibromyalgia symptoms, offering promising avenues for non-pharmacological management.

LRRK2 Inhibition by BIIB122 in Healthy Participants and Patients with Parkinson's Disease.

BACKGROUND: Leucine-rich repeat kinase 2 (LRRK2) inhibition is a promising therapeutic approach for the treatment of Parkinson's disease (PD). OBJECTIVE: The aim of this study was to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamics of the potent, selective, CNS-penetrant LRRK2 inhibitor BIIB122 (DNL151) in healthy participants and patients with PD. METHODS: Two randomized, double-blind, placebo-controlled studies were completed. The phase 1 study (DNLI-C-0001) evaluated single and multiple doses of BIIB122 for up to 28 days in healthy participants. The phase 1b study (DNLI-C-0003) evaluated BIIB122 for 28 days in patients with mild to moderate PD. The primary objectives were to investigate the safety, tolerability, and plasma pharmacokinetics of BIIB122. Pharmacodynamic outcomes included peripheral and central target inhibition and lysosomal pathway engagement biomarkers. RESULTS: A total of 186/184 healthy participants (146/145 BIIB122, 40/39 placebo) and 36/36 patients (26/26 BIIB122, 10/10 placebo) were randomized/treated in the phase 1 and phase 1b studies, respectively. In both studies, BIIB122 was generally well tolerated; no serious adverse events were reported, and the majority of treatment-emergent adverse events were mild. BIIB122 cerebrospinal fluid/unbound plasma concentration ratio was ~1 (range, 0.7-1.8). Dose-dependent median reductions from baseline were observed in whole-blood phosphorylated serine 935 LRRK2 (≤98%), peripheral blood mononuclear cell phosphorylated threonine 73 pRab10 (≤93%), cerebrospinal fluid total LRRK2 (≤50%), and urine bis (monoacylglycerol) phosphate (≤74%). CONCLUSIONS: At generally safe and well-tolerated doses, BIIB122 achieved substantial peripheral LRRK2 kinase inhibition and modulation of lysosomal pathways downstream of LRRK2, with evidence of CNS distribution and target inhibition. These studies support continued investigation of LRRK2 inhibition with BIIB122 for the treatment of PD. © 2023 Denali Therapeutics Inc and The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.

Influence of different Lachancea thermotolerans strains in the wine profile in the era of climate challenge.

The study performed sequential fermentations of red grape juice using several strains of Lachancea thermotolerans and one strain of Saccharomyces cerevisiae. Due to the new conditions imposed by climate change, wine acidity must be affected as well as the volatile profile. Non-Saccharomyces yeasts such as L. thermotolerans are real alternatives to soften the impact of climate change in winemaking. The L. thermotolerans strains included three commercially available strains and two wine-related natural isolates. L. thermotolerans showed significant statistical differences in basic chemical parameters such as lactic acid, malic acid, or ethanol concentrations as well as in the volatile profile. S. cerevisiae clearly produced some volatile compounds in higher amounts than the studied L. thermotolerans strains while others showed the opposite effect. Sequential fermentations involving any of the studied strains of L. thermotolerans with S. cerevisiae showed an increased volatile profile compared to the S. ceresisiae single fermentation, highlighting the synergic effect between the studied species.

Impact of rare yeasts in Saccharomyces cerevisiae wine fermentation performance: Population prevalence and growth phenotype of Cyberlindnera fabianii, Kazachstania unispora, and Naganishia globosa.

Saccharomyces cerevisiae is a highly fermentative species able to complete the wine fermentation. However, the interaction with other non-Saccharomyces yeasts can determine the fermentation performance of S. cerevisiae. We have characterised three rare non-Saccharomyces yeasts (Cyberlindnera fabianii, Kazachstania unispora and Naganishia globosa), studying their impact on S. cerevisiae fitness and wine fermentation performance. Using a wide meta-taxonomic dataset of wine samples, analysed through ITS amplicon sequencing, we show that about a 65.07% of wine samples contains Naganishia spp., a 27.21% contains Kazachstania spp., and only a 4.41% contains Cyberlindnera spp; in all cases with average relative abundances lower than 1% of total fungal populations. Although the studied N. globosa strain showed a limited growth capacity in wine, both K. unispora and C. fabianii showed a similar growth phenotype to that of S. cerevisiae in different fermentation conditions, highlighting the outstanding growth rate values of K. unispora. In mixed fermentations with S. cerevisiae, the three yeast species affected co-culture growth parameters and wine chemical profile (volatile compounds, polysaccharides and proteins). K. unispora DN201 strain presents an outstanding capacity to compete with S. cerevisiae strains during the first stage of wine fermentation, causing stuck fermentations in both synthetic and natural grape musts.

Mitochondrial Transfer as a Novel Therapeutic Approach in Disease Diagnosis and Treatment.

Mitochondrial dysfunction is a hallmark of numerous diseases, including neurodegenerative disorders, metabolic disorders, and cancer. Mitochondrial transfer, the transfer of mitochondria from one cell to another, has recently emerged as a potential therapeutic approach for restoring mitochondrial function in diseased cells. In this review, we summarize the current understanding of mitochondrial transfer, including its mechanisms, potential therapeutic applications, and impact on cell death pathways. We also discuss the future directions and challenges in the field of mitochondrial transfer as a novel therapeutic approach in disease diagnosis and treatment.

New Insights and Potential Therapeutic Interventions in Metabolic Diseases.

Endocrine homeostasis and metabolic diseases have been the subject of extensive research in recent years. The development of new techniques and insights has led to a deeper understanding of the mechanisms underlying these conditions and opened up new avenues for diagnosis and treatment. In this review, we discussed the rise of metabolic diseases, especially in Western countries, the genetical, psychological, and behavioral basis of metabolic diseases, the role of nutrition and physical activity in the development of metabolic diseases, the role of single-cell transcriptomics, gut microbiota, epigenetics, advanced imaging techniques, and cell-based therapies in metabolic diseases. Finally, practical applications derived from this information are made.

Non-immediate hypersensitivity reactions to iomeprol: Diagnostic value of skin tests and cross-reactivity with other iodinated contrast media.

BACKGROUND: Iodinated contrast media produce non-immediate hypersensitivity reactions (NIHR). The goal of this prospective study was to determine the utility of skin tests and the subsequent tolerance to negative skin-tested iodinated contrasts in patients with NIHR caused by iomeprol. METHODS: Prick and intradermal tests with iomeprol, iopamidol, iopromide, and iobitridol were performed in all patients. IV challenge with the causative contrast (iomeprol in 90%) was made if skin tests were negative. In case of a positive skin test with the causal contrast, or a positive challenge test with it, IV challenge test with an alternative, negative skin-tested contrast was performed in all patients. RESULTS: Skin tests were positive in 47.6% (20/42) of patients with NIHR induced by iomeprol. Of the 66 challenge tests performed with negative skin-tested iodinated contrasts, tolerance was confirmed in 35 (53%): 32 iomeron, 2 iobitridol, 1 iopamidol. Cross-reactivity between iomeprol and iopamidol was 22% (4/20 in patients with positive skin tests and 5/21 in patients with negative skin tests). CONCLUSIONS: Sensitivity of the skin tests was less than 50% NIHRs due to iomeprol, while the negative predictive value of skin tests in patients who tolerated challenges with alternative contrasts (mainly iopamidol) was 53% (35 tolerated out of 66 performed). The cross-reactivity between iomeprol and iopamidol is high.

Expectable diversity patterns in wine yeast communities.

Wine fermentations are dominated by Saccharomyces yeast. However, dozens of non-Saccharomyces yeast genera can be found in grape musts and in the early and intermediate stages of wine fermentation, where they co-exist with S. cerevisiae. The diversity of non-Saccharomyces species is determinant for the sensorial attributes of the resulting wines, both directly (by producing aroma impact compounds) and indirectly (modulating the performance of Saccharomyces). Many research groups worldwide are exploring the great diversity of wine yeasts to exploit their metabolic potential to improve wine flavor or to prevent wine spoilage. In this work, we share a new data set from a wide ITS amplicon survey of 272 wine samples, and we perform a preliminary exploration to build a catalogue of 242 fungal and yeast genera detectable in wine samples, estimating global figures of their prevalence and relative abundance patterns across wine samples. Thus, our mycobiome survey provides a broad measure of the yeast diversity potentially found in wine fermentations; we hope that the wine yeast research community finds it useful, and we also want to encourage further discussion on the advantages and limitations that meta-taxonomic studies may have in wine research and industry.

Molecular signatures of silencing suppression degeneracy from a complex RNA virus.

As genomic architectures become more complex, they begin to accumulate degenerate and redundant elements. However, analyses of the molecular mechanisms underlying these genetic architecture features remain scarce, especially in compact but sufficiently complex genomes. In the present study, we followed a proteomic approach together with a computational network analysis to reveal molecular signatures of protein function degeneracy from a plant virus (as virus-host protein-protein interactions). We employed affinity purification coupled to mass spectrometry to detect several host factors interacting with two proteins of Citrus tristeza virus (p20 and p25) that are known to function as RNA silencing suppressors, using an experimental system of transient expression in a model plant. The study was expanded by considering two different isolates of the virus, and some key interactions were confirmed by bimolecular fluorescence complementation assays. We found that p20 and p25 target a common set of plant proteins including chloroplastic proteins and translation factors. Moreover, we noted that even specific targets of each viral protein overlap in function. Notably, we identified argonaute proteins (key players in RNA silencing) as reliable targets of p20. Furthermore, we found that these viral proteins preferentially do not target hubs in the host protein interactome, but elements that can transfer information by bridging different parts of the interactome. Overall, our results demonstrate that two distinct proteins encoded in the same viral genome that overlap in function also overlap in their interactions with the cell proteome, thereby highlighting an overlooked connection from a degenerate viral system.

Psychophysiological and Psychosocial Profile of Patients Attending Drug Addiction Centers.

Drug treatment centres provide the highest level of rehab services for patients diagnosed with drug addictions. Most inpatient drug rehab programs focus on medical detox and mental health interventions. However, how to optimize the later remains a challenge. The aim of this study was to examine the psychophysiological and psychosocial profile of patients attending drug addiction centres in comparation with the general population. A total of 105 inpatient drug rehab patients and 50 participants from the general population were compared based on standardized psychophysiological and psychosocial measures. Results of this study suggest that patients attending drug addiction centers differ from general population in several different psychophysiological and psychosocial factors. Patients reported significantly lower levels of physical activity and increased sympathetic responsiveness, and significantly higher levels in loneliness, psychologically inflexibility and neuroticism. The results of this study highlight the importance of address healthy lifestyle behaviors such as sport practice and psychological variables such as loneliness, psychological (in)flexibility and neuroticism to improve current programs aim to prevent or reduce problematic drug consumptions.

Inflammation in COVID-19 and the Effects of Non-Pharmacological Interventions during the Pandemic: A Review.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused the coronavirus disease 2019 (COVID-19) pandemic that hit the health systems worldwide hard, causing a collapse of healthcare systems. One of the main problems of this new virus is the high inflammatory response it provokes, which is the cause of much of the symptoms. Different pharmacological approaches tried to stop the advance of the pandemic, but it seems that only vaccines are the solution. In this line, different nonpharmacological approaches have been made in order to improve symptomatology, contagion, and spread of COVID-19, the principal factors being the physical activity, nutrition, physiotherapy, psychology, and life patterns. The SARS-CoV-2 virus produces a disproportionate inflammatory response in the organism of the guest and causes complications in this that can end the life of the patient. It has been possible to see how different nonpharmacological interventions based on physical activity, nutritional, psychological, and physical therapy, and lifestyle changes can be functional tools to treat this inflammation. Thus, in the present review, we aim to provide an overview of the role of inflammation in COVID-19 and the nonpharmacological interventions related to it.

Etoricoxib-induced fixed drug eruption: Report of seven cases.

BACKGROUND: Fixed drug eruption (FDE) is a characteristic form of intraepidermal CD8+ T cell-mediated drug reaction, with repeated appearance of isolated or multiple skin lesions in the same location after receiving the offending drug. Non-steroidal anti-inflammatory drugs (NSAID) are the most common cause. Selective inhibitors of inducible cyclooxygenase 2 (COX-2) provoke a lesser degree of allergic or idiosyncratic adverse reactions than conventional NSAID, but they can cause skin reactions of variable severity. OBJECTIVE: Etoricoxib has been related to a variety of unusual skin reactions, including several reports of FDE. METHODS: We perfomed epicutaneous test to diagnose patients with suspected etoricoxib fixe drug rash due to clinical features and reproducibility on at least two occasions. RESULTS: We present seven new cases of etoricoxib-induced fixed drug eruption, with a diagnosis based on clinical presentation. This diagnosis was confirmed by an etoricoxib-positive lesional patch test in six cases and by a positive low-dose oral challenge in the other one. Two patients showed negative patch tests with celecoxib (10% in pet.) on the residual lesions, and oral tolerance was confirmed in one. CONCLUSION: To our knowledge, this is the largest series on FDE induced by etoricoxib reported to date.

A Novel Machine Learning-Based Methodology for Tool Wear Prediction Using Acoustic Emission Signals.

There is an increasing trend in the industry of knowing in real-time the condition of their assets. In particular, tool wear is a critical aspect, which requires real-time monitoring to reduce costs and scrap in machining processes. Traditionally, for the purpose of predicting tool wear conditions in machining, mathematical models have been developed to extract the information from the signal of sensors attached to the machines. To reduce the complexity of developing physical models, where an in-depth knowledge of the system being modelled is required, the current trend is to use machine-learning (ML) models based on data from the tool wear. The acoustic emission (AE) technique has been widely used to capture data from and understand the real-time condition of industrial assets such as cutting tools. However, AE signal interpretation and processing is rather complex. One of the most common features extracted from AE signals to predict the tool wear is the counts parameter, defined as the number of times that the amplitude of the signal exceeds a predefined threshold. A recurrent problem of this feature is to define the adequate threshold to obtain consistent wear prediction. Additionally, AE signal bandwidth is rather wide, and the selection of the optimum frequencies band for feature extraction has been pointed out as critical and complex by many authors. To overcome these problems, this paper proposes a methodology that applies multi-threshold count feature extraction at multiresolution level using wavelet packet transform, which extracts a redundant and non-optimal feature map from the AE signal. Next, recursive feature elimination is performed to reduce and optimize the vast number of predicting features generated in the previous step, and random forests regression provides the estimated tool wear. The methodology presented was tested using data captured when turning 19NiMoCr6 steel under pre-established cutting conditions. The results obtained were compared with several ML algorithms such as k-nearest neighbors, support vector machines, artificial neural networks and decision trees. Experimental results show that the proposed method can reduce the predicted root mean squared error by 36.53%.

High Potential of Pichia kluyveri and Other Pichia Species in Wine Technology.

The surfaces of grapes are covered by different yeast species that are important in the first stages of the fermentation process. In recent years, non-Saccharomyces yeasts such as Torulaspora delbrueckii, Lachancea thermotolerans, Metschnikowia pulcherrima, and Pichia kluyveri have become popular with regard to winemaking and improved wine quality. For that reason, several manufacturers started to offer commercially available strains of these non-Saccharomyces species. P. kluyveri stands out, mainly due to its contribution to wine aroma, glycerol, ethanol yield, and killer factor. The metabolism of the yeast allows it to increase volatile molecules such as esters and varietal thiols (aroma-active compounds), which increase the quality of specific varietal wines or neutral ones. It is considered a low- or non-fermentative yeast, so subsequent inoculation of a more fermentative yeast such as Saccharomyces cerevisiae is indispensable to achieve a proper fermented alcohol. The impact of P. kluyveri is not limited to the grape wine industry; it has also been successfully employed in beer, cider, durian, and tequila fermentation, among others, acting as a promising tool in those fermentation processes. Although no Pichia species other than P. kluyveri is available in the regular market, several recent scientific studies show interesting improvements in some wine quality parameters such as aroma, polysaccharides, acid management, and color stability. This could motivate yeast manufacturers to develop products based on those species in the near future.

Number of finishers and performance of age group women and men in long-distance running: comparison among 10km, half-marathon and marathon races in Oslo.

The aim of the present study was to examine the number of finishers and performance trends in 10 km, half-marathon and marathon races in Oslo. Data (total 115,725 finishers; women, n = 50,595; men, n = 65,130) from 10 km, half-marathon and marathon races in Oslo from 2008 to 2018 were analysed considering number, sex, age and running speed of finishers. The total men-to-women ratio was the smallest in the 10 km race (0.60) and the largest in the marathon (3.86) (p < 0.01, φ = 0.28). In both women and men, the slowest running speed was shown in the older age groups (p < 0.01). Based on the findings of the present study, it was concluded that relatively more women finished a 10 km and less a half-marathon and a marathon. Our results indicated that the sex difference in performance was attenuated in the longer race distances and older age groups.

Balanitis Xerotica Obliterans Associated With Differentiated Penile Intraepithelial Neoplasia and "Bramble-Bush" Elastic Fibers Accumulation: A Finding Not Specific for Penicillamine Intake.

We describe the association of balanitis xerotica obliterans and differentiated penile intraepithelial neoplasia (PeIN) with dermal elastosis and "bramble-bush" elastic fibers in a diabetic patient with no history of penicillamine intake. An 84-year-old man presented with urethral obstruction syndrome that required postectomy, meatotomy, and foreskin excision. Histopathological examination revealed changes of lichen sclerosus and differentiated PeIN. Orcein staining showed striking dermal elastosis and transepidermal elimination of elastic fibers. At higher magnification, multiple serrations and buds arising from the borders of the elastic fibers were observed giving the so-called bramble-bush appearance. Balanitis xerotica obliterans is considered a premalignant disease, and 14% of patients have evidence of PeIN. The differentiated subtype can be difficult to diagnose because of the minimal basal cell atypia, so a striking dermal elastosis, which may even occur before the neoplasm becomes invasive, may facilitate its diagnosis. The term "acquired perforating dermatosis" seems appropriate to describe those cases of perforating elastosis that occurs in adults with systemic diseases. The bramble-bush appearance of elastic fibers is not specific for penicillamine-induced elastopathy, and it may occur in other diseases, such as diabetes mellitus. This peculiar morphology of elastic fibers may be related to the enzymatic imbalance between matrix metalloproteinases and lysyl oxidase, an enzyme required for the cross-linking of elastic fibers.

Portable Biosensors for Psychophysiological Stress Monitoring of a Helicopter Crew.

This study aims to analyze the psychophysiological stress response of a helicopter crew using portable biosensors, and to analyze the psychophysiological stress response differences of experienced and non-experienced crew members. We analyzed 27 participants (33.89 ± 5.93 years) divided into two different flight maneuvers: a crane rescue maneuver: 15 participants (three control and 12 military) and a low-altitude maneuver: 12 participants (five control and seven military). Anxiety, rating of perceived exertion, subjective perception of stress, heart rate, blood oxygen saturation, skin temperature, blood lactate, cortical arousal, autonomic modulation, leg and hand strength, leg flexibility, spirometry, urine, and short-term memory were analyzed before and after both helicopter flight maneuvers. The maneuvers produced a significant increase in stress and effort perception, state of anxiety, and sympathetic modulation, as well as a significant decrease in heart rate, blood oxygen saturation, leg and inspiratory muscle strength, and urine proteins. The use of biosensors showed how a crane rescue and low-altitude helicopter maneuvers produced an anticipatory anxiety response, showing an increased sympathetic autonomic modulation prior to the maneuvers, which was maintained during the maneuvers in both experienced and non-experienced participants. The crane rescue maneuver produced a higher maximal heart rate and decreased pulmonary capacity and strength than the low-altitude maneuver. The psychophysiological stress response was higher in the experienced than in non-experienced participants, but both presented an anticipatory stress response before the maneuver.