Septins and K63 ubiquitin chains are present in separate bacterial microdomains during autophagy of entrapped Shigella.

During host cell invasion, Shigella escapes to the cytosol and polymerizes actin for cell-to-cell spread. To restrict cell-to-cell spread, host cells employ cell-autonomous immune responses including antibacterial autophagy and septin cage entrapment. How septins interact with the autophagy process to target Shigella for destruction is poorly understood. Here, we employed a correlative light and cryo-soft X-ray tomography (cryo-SXT) pipeline to study Shigella septin cage entrapment in its near-native state. Quantitative cryo-SXT showed that Shigella fragments mitochondria and enabled visualization of X-ray-dense structures (∼30 nm resolution) surrounding Shigella entrapped in septin cages. Using Airyscan confocal microscopy, we observed lysine 63 (K63)-linked ubiquitin chains decorating septin-cage-entrapped Shigella. Remarkably, septins and K63 chains are present in separate bacterial microdomains, indicating they are recruited separately during antibacterial autophagy. Cryo-SXT and live-cell imaging revealed an interaction between septins and LC3B-positive membranes during autophagy of Shigella. Together, these findings demonstrate how septin-caged Shigella are targeted for autophagy and provide fundamental insights into autophagy-cytoskeleton interactions.

Localization of all four ZnT zinc transporters in Dictyostelium and impact of ZntA and ZntB knockout on bacteria killing.

Professional phagocytes have developed an extensive repertoire of autonomous immunity strategies to ensure killing of bacteria. Besides phagosome acidification and the generation of reactive oxygen species, deprivation of nutrients and the lumenal accumulation of toxic metals are essential to kill ingested bacteria or inhibit the growth of intracellular pathogens. Here, we used the soil amoeba Dictyostelium discoideum, a professional phagocyte that digests bacteria for nutritional purposes, to decipher the role of zinc poisoning during phagocytosis of nonpathogenic bacteria and visualize the temporal and spatial dynamics of compartmentalized, free zinc using fluorescent probes. Immediately after particle uptake, zinc is delivered to phagosomes by fusion with 'zincosomes' of endosomal origin, and also by the action of one or more zinc transporters. We localized the four Dictyostelium ZnT transporters to endosomes, the contractile vacuole and the Golgi complex, and studied the impact of znt knockouts on zinc homeostasis. We show that zinc is delivered into the lumen of Mycobacterium smegmatis-containing vacuoles, and that Escherichia coli deficient in the zinc efflux P1B-type ATPase ZntA are killed faster than wild-type bacteria.

The ESCRT and autophagy machineries cooperate to repair ESX-1-dependent damage at the Mycobacterium-containing vacuole but have opposite impact on containing the infection.

Phagocytic cells capture and kill most invader microbes within the bactericidal phagosome, but some pathogens subvert killing by damaging the compartment and escaping to the cytosol. To prevent the leakage of pathogen virulence and host defence factors, as well as bacteria escape, host cells have to contain and repair the membrane damage, or finally eliminate the cytosolic bacteria. All eukaryotic cells engage various repair mechanisms to ensure plasma membrane integrity and proper compartmentalization of organelles, including the Endosomal Sorting Complex Required for Transport (ESCRT) and autophagy machineries. We show that during infection of Dictyostelium discoideum with Mycobacterium marinum, the ESCRT-I component Tsg101, the ESCRT-III protein Snf7/Chmp4/Vps32 and the AAA-ATPase Vps4 are recruited to sites of damage at the Mycobacterium-containing vacuole. Interestingly, damage separately recruits the ESCRT and the autophagy machineries. In addition, the recruitment of Vps32 and Vps4 to repair sterile membrane damage depends on Tsg101 but appears independent of Ca2+. Finally, in absence of Tsg101, M. marinum accesses prematurely the cytosol, where the autophagy machinery restricts its growth. We propose that ESCRT has an evolutionary conserved function to repair small membrane damage and to contain intracellular pathogens in intact compartments.

The relationship with the voices as a dialogical experience: The role of self-focused attention and dissociation.

OBJECTIVE: The purpose of this article was to study the relationship of self-focused attention and dissociation with the dialogical relationship persons diagnosed with psychosis have with their voices. METHOD: The DAIMON Scale was applied to 62 persons diagnosed with psychosis to measure the dialogical relationship with their voices, and the Cambridge depersonalization scale, the Tellegen absorption scale, and the self-focused attention scale. RESULTS: The results showed that the dialogical relationship with the voices was associated with high levels of self-focused attention (private and public), depersonalization, and absorption. It was also found that absorption mediated significantly between public self-focused attention and the dialogical relationship with the voices. CONCLUSIONS: The role of dissociation and self-focused attention in forming the dialogical relationship a person with psychosis has with the voices is discussed and approaches to treatment are suggested.

Mycobacterium marinum antagonistically induces an autophagic response while repressing the autophagic flux in a TORC1- and ESX-1-dependent manner.

Autophagy is a eukaryotic catabolic process also participating in cell-autonomous defence. Infected host cells generate double-membrane autophagosomes that mature in autolysosomes to engulf, kill and digest cytoplasmic pathogens. However, several bacteria subvert autophagy and benefit from its machinery and functions. Monitoring infection stages by genetics, pharmacology and microscopy, we demonstrate that the ESX-1 secretion system of Mycobacterium marinum, a close relative to M. tuberculosis, upregulates the transcription of autophagy genes, and stimulates autophagosome formation and recruitment to the mycobacteria-containing vacuole (MCV) in the host model organism Dictyostelium. Antagonistically, ESX-1 is also essential to block the autophagic flux and deplete the MCV of proteolytic activity. Activators of the TORC1 complex localize to the MCV in an ESX-1-dependent manner, suggesting an important role in the manipulation of autophagy by mycobacteria. Our findings suggest that the infection by M. marinum activates an autophagic response that is simultaneously repressed and exploited by the bacterium to support its survival inside the MCV.

Childhood memories of threatening experiences and submissiveness and its relationship to hallucination proneness and ideas of reference: The mediating role of dissociation.

Recent studies have emphasized the importance of childhood memories of threatening experiences and submissiveness in a diversity of psychological disorders. The purpose of this work was to study their specific relationship with hallucination proneness and ideas of reference in healthy subjects. The ELES scale for measuring memory of adverse childhood experiences, the DES-II scale for measuring dissociation, the LSHS-R scale for measuring hallucination proneness, and the REF for ideas of reference were applied to a sample of 472 subjects. A positive association was found between childhood memories of adverse experiences and hallucination proneness and ideas of reference, on one hand, and dissociation on the other. A mediation analysis showed that dissociation was a mediator between the memory of adverse childhood experiences and hallucination proneness on one hand, and ideas of reference on the other. When the role of mediator of the types of dissociative experiences was studied, it was found that absorption and depersonalization mediated between adverse experiences and hallucination proneness. However, this mediating effect was not found between adverse experiences and ideas of reference. The relationship between these last two variables was direct. The results suggest that childhood memories of adverse experiences are a relevant factor in understanding hallucination proneness and ideas of reference. Similarly, dissociation is a specific mediator between adverse childhood experiences and hallucination proneness.

Relationship between inner dialog and ideas of reference and the mediating role of dissociation.

Recent lines of research have begun to concentrate on internal dialogue and its relationship to a diversity of psychopathological phenomena present in psychotic disorders. This study was intended as a preliminary exploration of the relationship of internal dialogue, dissociation and ideas of reference. To do this, a sample of 318 students filled in an internal dialogue scale (the VISQ, McCarthy-Jones & Fernyhough), one for dissociation (DES-II, Carlson & Putnam) and another for ideas of reference (REF, Lenzenweger, Bennett & Lilenfeld). The results confirm the hypothesis posed in the sense that internal dialogue was positively associated with dissociation and with ideas of reference. A partial mediation effect of dissociation was also found between inner speech and ideas of reference. Lines of future research this study opens and its possible integration in a model on ideas of reference are discussed.

Dissociation and mindfulness in patients with auditory verbal hallucinations.

The very few studies relating mindfulness and dissociation have found a negative association between them (depersonalization and absorption). However, all of these studies have been done in nonclinical populations, and there are no data on the relationship between these variables in psychiatric patients with auditory hallucinations. This study was designed to study the relationship between mindfulness and the two dissociative variables, absorption and depersonalization, as well as their predictive power for the severity of auditory hallucinations and the distress they cause in a clinical population. A total of 55 psychiatric patients with hallucinations were given the following tests: the Mindful Attention Awareness Scale (K. Brown & R. Ryan, 2003), the Tellegen Absorption Scale (A. Tellegen & G. Atkinson, 1974), the Cambridge Depersonalization Scale (M. Sierra & G. E. Berrios, 2000), the Psychotic Symptom Rating Scales (G. Haddock, J. McCarron, N. Tarrier, & E. B. Faragher, 1999), and the Positive and Negative Syndrome Scale (S. R. Kay, L. A. Opler, & J.-P. Lindenmayer, 1988). A significant negative correlation was found between mindfulness and the dissociative variables and between mindfulness and the distress caused by the hallucinations. A positive correlation was found between absorption and distress caused by hallucinations and between depersonalization and the severity of hallucinations. Finally, the variable with the most predictive power for severity of the voices was depersonalization, and the variable with the most predictive power for distress caused by the voices was mindfulness. Interventions addressing training in mindfulness techniques could diminish the distress associated with hearing voices.

Relationship between childhood trauma, mindfulness, and dissociation in subjects with and without hallucination proneness.

The purpose of this study was to examine the relationship between childhood traumas, mindfulness, and dissociation (more specifically, absorption and depersonalization) in healthy subjects with and without hallucination proneness. A sample of 318 subjects was given the Launay-Slade Hallucination Scale-Revised (R. P. Bentall & P. Slade, 1985). From this sample, 2 groups were formed: one with high and the other with low hallucination proneness. Furthermore, all participants were given the Tellegen Absorption Scale (A. Tellegen & G. Atkinson, 1974), the Cambridge Depersonalization Scale (M. Sierra & G. E. Berrios, 2000), the Southampton Mindfulness Questionnaire (P. D. J. Chadwick et al., 2008), and the Trauma Questionnaire (J. R. E. Davidson, D. Hughes, & D. G. Blazer, 1990). The results showed that in the group with high hallucination proneness, there were significantly more subjects with traumatic experiences than in the group with low predisposition, although no significant difference in the mean number of traumatic experiences undergone in childhood was found between the 2 groups, although there was a trend toward significance. A correlation analysis showed a significant negative association between mindfulness on the one hand and absorption and depersonalization on the other. A positive relationship was also found between childhood traumas and absorption and depersonalization. Finally, multiple mediation analysis showed that the absorption and depersonalization variables acted as mediators between childhood traumas and hallucination proneness. We discuss the importance of the relationship between the variables studied and hallucination proneness and suggest some approaches for their treatment.

Relationship of absorption, depersonalisation, and self-focused attention in subjects with and without hallucination proneness.

INTRODUCTION: The purpose of this work was to study the relationship of absorption, depersonalisation, and self-focused attention in subjects prone to hallucination. METHODS: A sample of 218 healthy subjects was given the LSHS-R Hallucination Scale (Bentall & Slade, 1985). Three groups, subjects with high, medium, and low hallucination proneness, were formed from this sample. The Tellegen Absorption Scale (TAS; Tellegen & Atkinson, 1974), Cambridge Depersonalisation Scale (CDS; Sierra & Berrios, 2000), and Self-Absorption Scale (SAS; McKenzie & Hoyle, 2008) were also given to all the participants. The Metacognitions Questionnaire (MCQ-30; Wells & Cartwright-Hatton, 2004) was used as a covariant to control for the effects of emotional vulnerability on the dependent variables studied. RESULTS: The results showed that subjects highly prone to hallucinations had significantly higher absorption, depersonalisation, and self-focused attention than the subjects in the other two groups. A hierarchical regression analysis showed that absorption and depersonalisation predict hallucination proneness. CONCLUSIONS: The importance of the absorption, depersonalisation, and self-focused attention variables for understanding the aetiology of hallucinations is discussed in the Conclusions, where some approaches to its treatment are also suggested.

An automated microscopy workflow to study Shigella-neutrophil interactions and antibiotic efficacy in vivo.

Shigella are Gram-negative bacterial pathogens responsible for bacillary dysentery (also called shigellosis). The absence of a licensed vaccine and widespread emergence of antibiotic resistance has led the World Health Organisation (WHO) to highlight Shigella as a priority pathogen requiring urgent attention. Several infection models have been useful to explore the Shigella infection process; yet, we still lack information regarding events taking place in vivo. Here, using a Shigella-zebrafish infection model and high-content microscopy, we developed an automated microscopy workflow to non-invasively study fluorescently labelled bacteria and neutrophils in vivo. We applied our workflow to antibiotic-treated zebrafish, and demonstrate that antibiotics reduce bacterial burden and not neutrophil recruitment to the hindbrain ventricle. We discovered that nalidixic acid (a bactericidal antibiotic) can work with leukocytes in an additive manner to control Shigella flexneri infection and can also restrict dissemination of Shigella sonnei from the hindbrain ventricle. We envision that our automated microscopy workflow, applied here to study the interactions between Shigella and neutrophils as well as antibiotic efficacy in zebrafish, can be useful to innovate treatments for infection control in humans.

Interplay between septins and ubiquitin-mediated xenophagy during Shigella entrapment.

Septins are cytoskeletal proteins implicated in numerous cellular processes including cytokinesis and morphogenesis. In the case of infection by Shigella flexneri, septins assemble into cage-like structures that entrap cytosolic bacteria targeted by autophagy. The interplay between septin cage entrapment and bacterial autophagy is poorly understood. We used a correlative light and cryo-soft X-ray tomography (cryo-SXT) pipeline to study septin cage entrapment of Shigella in its near-native state. Septin cages could be identified as X-ray dense structures, indicating they contain host cell proteins and lipids consistent with their autophagy links. Airyscan confocal microscopy of Shigella-septin cages showed that septins and lysine 63 (K63)-linked ubiquitin chains are present in separate bacterial microdomains, suggesting they are recruited separately. Finally, Cryo-SXT and live-cell imaging revealed an interaction between septins and microtubule-associated protein light chain 3B (LC3B)-positive membranes during autophagy of Shigella. Collectively our data present a new model for how septin-caged Shigella are targeted to autophagy.

Acquisition of a large virulence plasmid (pINV) promoted temperature-dependent virulence and global dispersal of O96:H19 enteroinvasive Escherichia coli.

Enteroinvasive Escherichia coli (EIEC) and Shigella are closely related agents of bacillary dysentery. It is widely viewed that EIEC and Shigella species evolved from E. coli via independent acquisitions of a large virulence plasmid (pINV) encoding a type 3 secretion system (T3SS). Sequence Type (ST)99 O96:H19 E. coli is a novel clone of EIEC responsible for recent outbreaks in Europe and South America. Here, we use 92 whole genome sequences to reconstruct a dated phylogeny of ST99 E. coli, revealing distinct phylogenomic clusters of pINV-positive and -negative isolates. To study the impact of pINV acquisition on the virulence of this clone, we developed an EIEC-zebrafish infection model showing that virulence of ST99 EIEC is thermoregulated. Strikingly, zebrafish infection using a T3SS-deficient ST99 EIEC strain and the oldest available pINV-negative isolate reveals a separate, temperature-independent mechanism of virulence, indicating that ST99 non-EIEC strains were virulent before pINV acquisition. Taken together, these results suggest that an already pathogenic E. coli acquired pINV and that virulence of ST99 isolates became thermoregulated once pINV was acquired. IMPORTANCE Enteroinvasive Escherichia coli (EIEC) and Shigella are etiological agents of bacillary dysentery. Sequence Type (ST)99 is a clone of EIEC hypothesized to cause human disease by the recent acquisition of pINV, a large plasmid encoding a type 3 secretion system (T3SS) that confers the ability to invade human cells. Using Bayesian analysis and zebrafish larvae infection, we show that the virulence of ST99 EIEC isolates is highly dependent on temperature, while T3SS-deficient isolates encode a separate temperature-independent mechanism of virulence. These results indicate that ST99 non-EIEC isolates may have been virulent before pINV acquisition and highlight an important role of pINV acquisition in the dispersal of ST99 EIEC in humans, allowing wider dissemination across Europe and South America.

Dissociative experiences as mediators between childhood trauma and auditory hallucinations.

The purpose of this work was to study the relationship between reported traumatic experiences in childhood and positive psychotic symptoms. We hypothesized that dissociative experiences were potential mediators between childhood trauma and hallucinations, but not delusions. The sample comprised 71 patients diagnosed with psychoses. They were assessed with the Dissociative Experiences Scale (DES-II; Carlson & Putnam, 1993), a questionnaire on trauma (TQ; Davidson, Hughes, & Blazer, 1990), and the Positive and Negative Syndrome Scale (PANSS; Kay, Opler, & Lindenmayer, 1988) delusions and hallucinations items. The results showed that childhood trauma was positively associated with the dissociation scale scores (r = .40) and also the hallucination (r = .36) and delusions scale scores (r = .32). Furthermore, it was demonstrated that the dissociation variable was a potential mediator between childhood trauma and hallucinations, but not between childhood trauma and delusions. Of the 3 DES-II factors, only depersonalization showed a mediating relationship between childhood trauma and hallucinations. The main conclusion is that the impact of childhood trauma on hallucinations may not simply be direct, but mediated by dissociative experiences, especially depersonalization. Clinical implications are also briefly discussed.

Job satisfaction and depression in the Spanish Society of Periodontology and Research (SEPA) members, and their relation to the burnout syndrome. Creation of a structural model.

OBJECTIVE: This study is aimed at getting to know the existing relationship between the dimensions of the burnout syndrome and job satisfaction, on one hand, and depressive feelings on the other through the creation of a structural model aimed at relating all these concepts on a sample of Spanish periodontists. STUDY DESIGN: The initial sample comprised 284 individuals, who represented 20% of the members of the Spanish Society of Periodontology and Research (www.SEPA.es). These individuals were chosen randomly by means of stratified sampling with proportional affixation by their autonomous community of residence. All participants were sent by post the MBI, CET and job-satisfaction questionnaires. The software package used for data analysis was LISREL v. 8.7 by checking models of structural equations so as to prove the proposed model's adjustment. RESULTS: The total number of answered questionnaires was 170 (59.85%). A positive relation was observed between emotional tiredness and depersonalization and depression. However, this dimension correlated negatively with job satisfaction and self-realization. CONCLUSIONS: The obtained results show that, in this sample of periodontists, job satisfaction acts as a modulator in the transition from emotional tiredness to depression.

Spanish adaptation of the scale of psychological empowerment in the workplace.

The objective of this study is to adapt and translate into Spanish Spreitzer's Psychological Empowerment Scale (1995a). A process of translation and reverse-translation was applied to the scale's items, whose psychometric properties were then examined using a sample of 272 professional nurses at public hospitals in the province of Seville. The data were subjected to confirmatory factor analysis. The significance of the factor loadings demonstrated the need to create a new model eliminating one item. The 11-item model was shown to possess adequate construct validity and internal consistency. The results confirm the original, four-factor structure obtained by Spreitzer, with the exception of item 10, and support the utilization of the Spanish version of this scale in the workplace. Future research should more extensively investigate its construct validity, and test the nomological network of the operationalized construct within the field of psychological well-being and in the context of the workplace.

Elevated monocyte distribution width in trauma: An early cellular biomarker of organ dysfunction.

INTRODUCTION: Traumatic injury elicits an inflammatory response such as the one occurring during systemic infection. Monocyte distribution width (MDW) has been found to distinguish sepsis in a pool of patients with suspected infection. We hypothesized that an elevated MDW in trauma patients would be associated with the development of multiple organ dysfunction syndrome (MODS) and an increased mortality. MATERIALS AND METHODS: Observational study in a dedicated trauma Intensive Care Unit (ICU) in Madrid during 2019-2020. Patients were classified according to their first MDW value on admission, as greater or lesser than 21 U. Clinical data was obtained and univariate and multivariate analysis were realized, as well as a test performance analysis. RESULTS: 354 patients were studied, with a median age of 46 years, 78% male. Half presented with severe trauma ISS > 15, mostly with a blunt mechanism of injury. A MDW ≥ 21 U on admission was found in 17% of cases. These patients were more likely to present with hemodynamic instability and MODS. They had a higher length of stay (3.8 vs 2 days) and higher mortality (21 vs 5%) compared to the low MDW group. These findings remained statistically significant in the multivariate analysis, with an OR 4.6 (IC 95% 1.7-12) for MODS and 3.1 (IC 95% 1.2-8.3) for mortality. CONCLUSIONS: In trauma patients, a MDW ≥ 21 U on admission was independently associated with a greater risk of MODS, a higher mortality and a higher length of stay. This biomarker could be useful in predicting severity in the initial evaluation of trauma patients.

Emerging technologies and infection models in cellular microbiology.

The field of cellular microbiology, rooted in the co-evolution of microbes and their hosts, studies intracellular pathogens and their manipulation of host cell machinery. In this review, we highlight emerging technologies and infection models that recently promoted opportunities in cellular microbiology. We overview the explosion of microscopy techniques and how they reveal unprecedented detail at the host-pathogen interface. We discuss the incorporation of robotics and artificial intelligence to image-based screening modalities, biochemical mapping approaches, as well as dual RNA-sequencing techniques. Finally, we describe chips, organoids and animal models used to dissect biophysical and in vivo aspects of the infection process. As our knowledge of the infected cell improves, cellular microbiology holds great promise for development of anti-infective strategies with translational applications in human health.

[Minimal important clinical difference in health-related quality of life.] / Diferencia mínima clínica importante en calidad de vida relacionada con la salud.

Una de las limitaciones para el uso de los Cuestionarios de Calidad de Vida Relacionada con la Salud y de los resultados reportados por pacientes es la dificultad de interpretar los valores obtenidos. La diferencia mínima clínica importante es una medida que ayuda a comprender los resultados de estos cuestionarios y valorar la relevancia clínica del efecto conseguido por la intervención realizada. En este trabajo revisamos el concepto de diferencia mínima clínica importante, describimos los métodos utilizados para su obtención y exponemos sus dificultades, limitaciones y aplicabilidad. Dentro de los resultados reportados por pacientes y, particularmente, en los de calidad de vida, la diferencia mínima clínica importante es una herramienta que ayuda a los clínicos a utilizar correctamente las escalas de medida e interpretar el efecto de las intervenciones. Con este artículo esperamos facilitar la implantación y uso de la diferencia mínima clínica importante y los Cuestionarios de Calidad de Vida Relacionada con la Salud en la práctica habitual con nuestros pacientes.

One of the limitations for the use of Health-related Quality of Life Questionnaires and the results reported by patients is the difficulty of interpreting the values obtained. The minimal important clinical difference is a measure that helps to understand the results of these questionnaires and assess the clinical relevance of the effect achieved by the intervention performed. In this paper, we review the concept of minimal important clinical difference, describe the methods used to obtain it, and expose its difficulties, limitations, and applicability. Within the results reported by patients, and particularly in quality of life, the minimal important clinical difference is a tool that helps clinicians to correctly use the measurement scales and interpret the effect of the interventions. With this article, we hope to facilitate the implementation and use of the minimal important clinical difference and the Health-related Quality of Life Questionnaires in routine practice with our patients.