Current and novel approaches in yeast cell death research.

The study of cell death mechanisms in fungi, particularly yeasts, has gained substantial interest in recent decades driven by the potential for biotechnological advancements and therapeutic interventions. Examples include the development of robust yeast strains for industrial fermentations and high-value compound production, novel food preservation strategies against spoilage yeasts, and the identification of targets for treating fungal infections in the clinic. In this review, we discuss a wide range of methods to characterize cellular alterations associated with yeast cell death, noting the advantages and limitations. We describe assays to monitor reversible events versus those that mark a commitment to cell death (point-of-no-return), as these distinctions are important to decipher the underlying regulatory mechanisms. Several well-known challenges remain, including the varied susceptibilities to death within a cell population and the delineation of detailed cell death mechanisms. The identification and characterization of morphologically distinct subsets of dying yeast cells within dynamic yeast populations provides opportunities to reveal novel vulnerabilities and survival mechanisms. Elucidating the intricacies of yeast regulated cell death (yRCD) will contribute to the advancement of scientific knowledge and foster breakthrough discoveries with broad-ranging implications.

The fungicide cymoxanil impairs respiration in Saccharomyces cerevisiae via cytochrome c oxidase inhibition.

Cymoxanil (CYM) is a widely used synthetic acetamide fungicide, but its biochemical mode of action remains elusive. Since CYM inhibits cell growth, biomass production, and respiration in Saccharomyces cerevisiae, we used this model to characterize the effect of CYM on mitochondria. We found it inhibits oxygen consumption in both whole cells and isolated mitochondria, specifically inhibiting cytochrome c oxidase (CcO) activity during oxidative phosphorylation. Based on molecular docking, we propose that CYM blocks the interaction of cytochrome c with CcO, hampering electron transfer and inhibiting CcO catalytic activity. Although other targets cannot be excluded, our data offer valuable insights into the mode of action of CYM that will be instrumental in driving informed management of the use of this fungicide.

Psycho-emotional intervention with parents of very preterm babies during the first year: A single-arm pilot study.

Studies of intervention programs that aim to improve the emotional state of parents of children admitted to the neonatal intensive care units (NICU) are scarce in Spain. The aims of this single-arm pilot study are to get to know the emotional profile of parents of high-risk preterm newborns, and to explore parents' patterns of emotional well-being before and after a psychological program called the Parental Empowerment Program, to increase parental readiness levels. The sample was made up of 100 parents (50 couples) who participated in the program. Measurements were taken of post-traumatic stress, depression, and resilience at 1 month and 12 months. Repeated measurements and dyadic data analyses were performed. One month after the birth of the baby and prior to the start of the program, mothers show more symptoms of stress and depression than fathers. After the intervention, both parents experienced improvements in their mood levels. The evidence obtained seems to show that high resilience levels and low post-traumatic stress symptoms are associated with reduced depression levels after implementing the program. However, the heterogeneity of the responses obtained, the observed associations between stress, resilience, and maternal depression, along with the reciprocal influence between maternal and paternal depression 1 year after the intervention, highlight the need for a more in-depth exploration of the interplay between risk and protective factors in this population. Despite the identified potential threats to validity, further work in this direction is recommended, including the implementation of clinical trials to demonstrate intervention efficacy. The adaptation of the parents' mutual emotional adjustment at each stage would allow them to participate more actively in the baby's care.

Enhancement of Acetate-Induced Apoptosis of Colorectal Cancer Cells by Cathepsin D Inhibition Depends on Oligomycin A-Sensitive Respiration.

Colorectal cancer (CRC) is a leading cause of death worldwide. Conventional therapies are available with varying effectiveness. Acetate, a short-chain fatty acid produced by human intestinal bacteria, triggers mitochondria-mediated apoptosis preferentially in CRC but not in normal colonocytes, which has spurred an interest in its use for CRC prevention/therapy. We previously uncovered that acetate-induced mitochondrial-mediated apoptosis in CRC cells is significantly enhanced by the inhibition of the lysosomal protease cathepsin D (CatD), which indicates both mitochondria and the lysosome are involved in the regulation of acetate-induced apoptosis. Herein, we sought to determine whether mitochondrial function affects CatD apoptotic function. We found that enhancement of acetate-induced apoptosis by CatD inhibition depends on oligomycin A-sensitive respiration. Mechanistically, the potentiating effect is associated with an increase in cellular and mitochondrial superoxide anion accumulation and mitochondrial mass. Our results provide novel clues into the regulation of CatD function and the effect of tumor heterogeneity in the outcome of combined treatment using acetate and CatD inhibitors.

Novel yeast-based biosensor for environmental monitoring of tebuconazole.

Due to increasing demand for high and stable crop production, human populations are highly dependent on pesticide use for growing and storing food. Environmental monitoring of these agrochemicals is therefore of utmost importance, because of their collateral effects on ecosystem and human health. Even though most current-use analytical methods achieve low detection limits, they require procedures that are too complex and costly for routine monitoring. As such, there has been an increased interest in biosensors as alternative or complementary tools to streamline detection and quantification of environmental contaminants. In this work, we developed a biosensor for environmental monitoring of tebuconazole (TEB), a common agrochemical fungicide. For that purpose, we engineered S. cerevisiae cells with a reporter gene downstream of specific promoters that are expressed after exposure to TEB and characterized the sensitivity and specificity of this model system. After optimization, we found that this easy-to-use biosensor consistently detects TEB at concentrations above 5 µg L-1 and does not respond to realistic environmental concentrations of other tested azoles, suggesting it is specific. We propose the use of this system as a complementary tool in environmental monitoring programs, namely, in high throughput scenarios requiring screening of numerous samples. KEY POINTS: • A yeast-based biosensor was developed for environmental monitoring of tebuconazole. •The biosensor offers a rapid and easy method for tebuconazole detection ≥ 5 µg L-1. •The biosensor is specific to tebuconazole at environmentally relevant concentrations.

The Methodological Quality Scale (MQS) for intervention programs: validity evidence.

Introduction: A wide variety of instruments are used when assessing the methodological quality (MQ) of intervention programs. Nevertheless, studies on their metric quality are often not available. In order to address this shortcoming, the methodological quality scale (MQS) is presented as a simple and useful tool with adequate reliability, validity evidence, and metric properties. Methods: Two coders independently applied the MQS to a set of primary studies. The number of MQ facets was determined in parallel analyses before performing factor analyses. For each facet of validity obtained, mean and standard deviation are presented jointly with reliability and average discrimination. Additionally, the validity facet scores are interpreted based on Shadish, Cook, and Campbell's validity model. Results and discussion: An empirical validation of the three facets of the MQ (external, internal, and construct validity) and the interpretation of the scores were obtained based on a theoretical framework. Unlike other existing scales, MQS is easy to apply and presents adequate metric properties. In addition, MQ profiles can be obtained in different areas of intervention using different methodologies and proves useful for both researchers doing meta-analysis and for evaluators and professionals designing a new intervention.

Methodological procedure based on quantitizing/liquefying: a case study to assess work climate in an emergency department.

Introduction: In the assessment of health organizations, results-based indicators are mainly used, with no consideration of internal work dynamics. This type of assessment forfeits much of the rich, useful information needed to make decisions on improving the organization. In order to address this, a rigorous procedure based on mixed methods is laid out here on gathering, analyzing, and interpreting data associated with the implementation process. Methods: A 55-year-old doctor was selected at random from among the staff who volunteered to be interviewed at the emergency department at a public hospital located in southern Spain for an interview. Qualitative data obtained from the in-depth interview (indirect observation) were progressively systematized (liquefied and quantitized) based on a theoretical framework until a code matrix was obtained, without losing or distorting any information. Afterwards, data quality was controlled using Cohen's kappa (κ) coefficient. A quantitative polar coordinate analysis was then carried out using the free software HOISAN (v. 1.6.3.3) to obtain robust results, vectorizing the relationships between codes and specifying whenever such relationships were statistically significant (and if they resulted in behavior activation or inhibition). Finally, a supplementary quantitative and qualitative assessment was carried out. Results and discussion: The proposed method was applied to the needs assessment of teams in order to evaluate that work climate in the hospital's emergency department Health Services of a hospital. Data quality control yielded an adequate result (κ = 0.82). Significant activation and inhibition of behaviors occurred, both prospectively and retrospectively. For instance, We seek to understand the needs of our clients and We readily adapt to new circumstances showed a significant activation (vector length = 3.43, p < 0.01) both prospectively (Zsum = 0.48) and retrospectively (Zsum = 3.4).An adequate method to obtain detailed information about group dynamics in a work environment is presented, based on an in-depth interview. Practical applications for implementations to improve the functioning of organizations are presented.

Monitoring yeast regulated cell death: trespassing the point of no return to loss of plasma membrane integrity.

Acetic acid and hydrogen peroxide are the most common stimuli to induce apoptosis in yeast. The initial phase of this cell death process is characterized by the maintenance of plasma membrane integrity in cells that had already lost their viability. As loss of plasma membrane integrity is typically assessed by staining with propidium iodide (PI) after exposure of cells to a stimulus and cell viability is determined 48 h after plating, the percentage of cells with compromised plasma membrane integrity and c.f.u. counts often do not correlate. Herein, we developed a simple method to explore at what point after an apoptotic stimulus and plating cells do non-viable cells die as result of plasma membrane disruption, i.e., when cells surpass the point-of-no-return and undergo a secondary necrosis. The method consisted in washing cells and re-suspending them in stimulus-free medium after acetic acid and hydrogen peroxide treatments, to mimic transfer to plating, and then assessing plasma membrane integrity through PI staining. We show that, after the stimuli are removed, cells that had lost proliferative capacity but still maintained plasma membrane integrity continue the cell death process and later lose plasma membrane integrity when progressing to secondary necrosis. After exposure to hydrogen peroxide, cells undergo secondary necrosis preceded by Nhp6Ap-GFP cytosolic localization, in contrast to acetic acid exposure, where Nhp6Ap-GFP cytosolic localization mainly occurs simultaneously with an earlier emergence of secondary necrosis. In conclusion, the developed method allows monitoring the irreversible loss of plasma membrane integrity of dying apoptotic cells after the point-of-no-return is trespassed, and better characterize the process of secondary necrosis after apoptosis.

Emotional Self-Regulation in Everyday Life: A Systematic Review.

Emotional self-regulation in childhood and adolescence constitutes a growing interest in the scientific community, highlighting in recent years the need to observe its development in their daily life. Therefore, the objective of this systematic review is to characterize publications referring to the development of emotional self-regulation of people under 18 years-old, in natural contexts. Based on the PRISMA guidelines, searches are carried out in the Web of Science, Scopus and PsycINFO databases, and in Google Scholar until May 2020. After reviewing the full text of 376 publications, 14 works are selected that are observed in their extrinsic, substantive and methodological characteristics based on the GREOM and MQCOM guidelines, by two independent evaluators. Most of the studies correspond to the last 20 years, increasing the interest in observing older children, in interaction with adults and/or in different cultures. They apply mixed methodologies, not always ascribing to a low intensity design. Strengths are observed regarding the collection and analysis of the quality of the data; and weaknesses related to the failure to record the duration and sequence of behaviors, highlighting the use of guidelines as guides for future research.

Acetic acid triggers cytochrome c release in yeast heterologously expressing human Bax.

Proteins of the Bcl-2 protein family, including pro-apoptotic Bax and anti-apoptotic Bcl-xL, are critical for mitochondrial-mediated apoptosis regulation. Since yeast lacks obvious orthologs of Bcl-2 family members, heterologous expression of these proteins has been used to investigate their molecular and functional aspects. Active Bax is involved in the formation of mitochondrial outer membrane pores, through which cytochrome c (cyt c) is released, triggering a cascade of downstream apoptotic events. However, when in its inactive form, Bax is largely cytosolic or weakly bound to mitochondria. Given the central role of Bax in apoptosis, studies aiming to understand its regulation are of paramount importance towards its exploitation as a therapeutic target. So far, studies taking advantage of heterologous expression of human Bax in yeast to unveil regulation of Bax activation have relied on the use of artificial mutated or mitochondrial tagged Bax for its activation, rather than the wild type Bax (Bax α). Here, we found that cell death could be triggered in yeast cells heterologoulsy expressing Bax α with concentrations of acetic acid that are not lethal to wild type cells. This was associated with Bax mitochondrial translocation and cyt c release, closely resembling the natural Bax function in the cellular context. This regulated cell death process was reverted by co-expression with Bcl-xL, but not with Bcl-xLΔC, and in the absence of Rim11p, the yeast ortholog of mammalian GSK3ß. This novel system mimics human Bax α regulation by GSK3ß and can therefore be used as a platform to uncover novel Bax regulators and explore its therapeutic modulation.

Saccharomyces cerevisiae Cells Lacking the Zinc Vacuolar Transporter Zrt3 Display Improved Ethanol Productivity in Lignocellulosic Hydrolysates.

Yeast-based bioethanol production from lignocellulosic hydrolysates (LH) is an attractive and sustainable alternative for biofuel production. However, the presence of acetic acid (AA) in LH is still a major problem. Indeed, above certain concentrations, AA inhibits yeast fermentation and triggers a regulated cell death (RCD) process mediated by the mitochondria and vacuole. Understanding the mechanisms involved in AA-induced RCD (AA-RCD) may thus help select robust fermentative yeast strains, providing novel insights to improve lignocellulosic ethanol (LE) production. Herein, we hypothesized that zinc vacuolar transporters are involved in vacuole-mediated AA-RCD, since zinc enhances ethanol production and zinc-dependent catalase and superoxide dismutase protect from AA-RCD. In this work, zinc limitation sensitized wild-type cells to AA-RCD, while zinc supplementation resulted in a small protective effect. Cells lacking the vacuolar zinc transporter Zrt3 were highly resistant to AA-RCD, exhibiting reduced vacuolar dysfunction. Moreover, zrt3Δ cells displayed higher ethanol productivity than their wild-type counterparts, both when cultivated in rich medium with AA (0.29 g L-1 h-1 versus 0.11 g L-1 h-1) and in an LH (0.73 g L-1 h-1 versus 0.55 g L-1 h-1). Overall, the deletion of ZRT3 emerges as a promising strategy to increase strain robustness in LE industrial production.

Predictive Factors of Mental Health in Survivors of Intimate Partner Violence in Chile.

Studies about violence against women specific to the Chilean population are scarce. As a result, government treatment programs lack a local perspective. Predictor variables were analyzed in the mental health of Chilean women who have survived intimate partner abuse. Two hundred and two women who made regular visits to public Women's Centers participated in the study; on average, they had survived 11 years of abuse. Logistic regression analyses were conducted to determine what variable/s in the history of violence best predicted the mental health variables. Among other protective factors, an increase in both resilience and the time since the last violent episode yielded a reduction in levels of general psychological distress (B = -1.836, p < .001 and B = 1.117, p < .001 respectively), post-traumatic stress disorder (B = -1.243, p = .002 and B = 1.221, p < .001 respectively), and depression (B = -1.822, p < .001 and B = 1.433, p < .001 respectively). The study also noted risk factors such as a high level of additional stressors, which in turn led to increased levels of general psychological distress (B = 1.007, p = .005), post-traumatic stress disorder (B = 0.928, p = .013), and depression (B = 1.061, p = .016). The Women's Center is the place where women feel most supported. To improve the effectiveness of treatments at these centers and aid in the recovery of women who have suffered from intimate partner violence, the predictive factors significantly related to mental health should be taken into account. This means prioritizing cases where the last episode of violence was more recent, addressing additional stressors, and promoting resilience.

The PALliative MUlticenter Study in Intensive Care (PalMuSIC). Results From a Multicenter Study Addressing Frailty and Palliative Care Interventions in Intensive Care Units in Portugal.

Objective: Frailty is a clinically recognizable state of increased vulnerability common in critical medicine. When underrecognized, it may lead to invasive treatments that do not serve the patients' best interest. Our aim was to evaluate the use of both palliative care consultation and invasive interventions in frail patients admitted to Intensive Care Units in Portugal. Methods: This was a prospective, observational study. All consecutive adult patients admitted for more than 24 h, over a 15-day period were enrolled. Twenty-three Portuguese Intensive Care Units were included. Informed consent was obtained from all patients or their surrogate. The doctor and nurse in charge calculated the Clinical Frailty Score as well as the reference family member Results: A total of 335 patients were included in the study (66% male). Mean age was 63.2 ± 16.8 and SAPS II score was 41.8 ± 17.4. Mean Clinical Frailty Score value was 3.5 ± 1.7. Frailty prevalence (mean score ≥ 5) was 20.9%. Frail patients were offered organ support therapy (64,3% invasive mechanical ventilation; 24,3% renal replacement therapy; 67,1% vasopressors) more often than non-frail patients. Nevertheless, limitation of therapeutic effort or a do not resuscitate order (p < 0.001) were more common in frail patients. Mortality rate by 6 months was higher among frail patients (50% vs. 32.3%, p < 0.001). Palliative Care was offered to only 15% of frail patients (3.9% overall). Conclusions: The authors suggest that palliative care should be universally consulted once frailty is identified in critical patients.

Severe Acute Respiratory Syndrome Coronavirus 2 Antibodies Among Healthcare Workers After Vaccine Administration in an Intensive Care Unit.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and coronavirus disease 2019 (COVID-19) emerged in China in December 2019. Healthcare workers (HCWs) are one of the high-risk groups of infection and knowledge of the seroprevalence of SARS-CoV-2 antibodies among this class is very important, not only to understand the spread of COVID-19 among health institutions but also to assess the success of public health interventions. The objective of this prospective study was to determine the seroprevalence of COVID-19 immunoglobulin G (IgG) antibodies after vaccine administration and assess the symptomatology associated with the number of IgG antibodies. A total of 75 HCWs from an intensive care unit were studied three and six months after the second administration of the COVID-19 vaccine. They were divided into three groups: IgG antibodies between 4,160 and 6,350 (group one), greater than 6,350 (group two), and less than 4,160 (group three). After the first administration of the vaccine, 80% had symptoms in both groups one and two, whereas only 13.8% had symptoms in group three. After the second dose of the vaccine, all elements of group one and 80% of group two developed symptoms, but only 40% of group three manifested symptoms. With the exception of one, all professionals showed a decrease in the number of IgG antibodies from three to six months. Our findings show that professionals with a higher number of IgG antibodies had more symptoms and that these rapidly declined over the three-to-six-month period.

From randomized control trial to mixed methods: a practical framework for program evaluation based on methodological quality / Del ensayo controlado aleatorizado a los métodos mixtos: un marco práctico para la evaluación de programas basado en la calidad metodológica

The evidence used when making decisions about the design, implementation and evaluation in intervention programs should be methodologically sound. Depending on the context of the intervention, different methodologies may apply. Nonetheless, the intervention context is often unstable and, to adapt to changing circumstances, it sometimes becomes necessary to modify the original plan. The framework proposed herein draws on approaches that can be considered two extremes of a continuum (experimental/quasi-experimental designs and studies based on observational methodology). In unstable intervention context conditions, this enables decisions from a methodological quality approach regarding design, measurement, and analysis. Structural dimensions, i.e., units (participants, users), treatment (program activities), outcomes (results, including decisions about the instruments to use and data gathering), setting (implementation context) and time will be detailed as part of the practical framework. The present study aims to specify the degree of correspondence/complementarity between components in these structural dimensions of a program evaluation from a practical complementarity perspective based on methodological quality.(AU)

La evidencia utilizada al tomar decisiones sobre el diseño, implementación y evaluación en los programas de intervención debe ser metodológicamente sólida. Dependiendo del contexto de la intervención, se pueden aplicar diferentes metodologías. Sin embargo, el contexto de la intervención es a menudo inestable y, para adaptarse a las circunstancias cambiantes, en algunas ocasiones se hace necesario modificar el plan original. El marco propuesto en este documento se basa en enfoques que pueden considerarse dos extremos de un continuo (diseños experimentales / cuasiexperimentales y estudios basados ​​en metodología observacional). En condiciones de contexto de intervención inestable, esto permite tomar decisiones desde un enfoque de calidad metodológica en cuanto a diseño, medición y análisis. Las dimensiones estructurales, i.e., las unidades (participantes, usuarios), el tratamiento (actividades del programa), los resultados (incluidas las decisiones sobre los instrumentos a utilizar y la recopilación de datos), el entorno (contexto de implementación) y el tiempo se detallarán como parte del marco práctico. El presente estudio tiene como objetivo especificar el grado de correspondencia / complementariedad entre componentes en estas dimensiones estructurales de la evaluación de un programa desde una perspectiva de complementariedad práctica basada en la calidad metodológica.(AU)

A Training Intervention through a 360° Multisource Feedback Model.

Physicians and other health sciences professionals need continuous training, not only in technical aspects of their activity but also in nontechnical, transversal competencies with a cost-efficient impact on the proper functioning of healthcare. The objective of this paper is to analyze the behavioral change among health professionals at a large public hospital following a training intervention on a set of core nontechnical competencies: Teamwork, Adaptability-Flexibility, Commitment-Engagement, Results Orientation, and Leadership Skills for Supervisors. The 360° Multisource Feedback (MSF) model was applied using three sources of information: supervisors, co-workers, and the workers themselves (self-assessment). A quasi-experimental pretest-post-test single-group design with two points in time was utilized. The training intervention improved the scores of only one of the trained competencies-the "Results Orientation" competency-although the scores were slightly inflated. Moreover, significant discrepancies were detected between the three sources, with supervisors awarding the highest scores. The magnitude of behavioral change was related to certain sociodemographic and organizational variables. The study was not immune to the ceiling effect, despite control measures aimed at avoiding it. The empirical evidence suggests that the 360° MSF model must be maintained over time to enhance and reinforce an evaluation culture for better patient care.

Effect of a Ketogenic Diet on the Nutritional Parameters of Obese Patients: A Systematic Review and Meta-Analysis.

The effect of a ketogenic diet (KD) on biochemical parameters and nutritional status has been the subject of debate over the years, as several randomized clinical trials (RCTs) obtained different results. METHOD: A systematic review and random-effects meta-analysis of RCTs comparing KD with a balanced diet was performed by means of a search of PubMed, Cochrane Library, Scopus, and Web of Science. Trials where the method for measuring the response variables was unclear, those that considered pathologies other than chronic non-communicable diseases and those with participants receiving pharmacological treatment for obesity were excluded from the comparison. RESULTS: Of the studies included in the meta-analysis, no statistically significant standardized mean differences were observed for body mass index (BMI) (d = -0.457, p = 0.359), total cholesterol, COL-T (d = 0.230, p = 0.591), high-density lipoprotein, HDL (d = -0.028, p = 0.934), low-density lipoprotein, LDL (d = 0.528, p = 0.173), or triglycerides, TG (d = -0.283, p = 0.222), with high values of heterogeneity. The percentage of women included in the studies is a significant moderating variable in terms of BMI ratio (z = -6.68, p < 0.001) and TG (z = -2.27, p = 0.023). CONCLUSION: A KD shows no more benefits on nutritional parameters than a balanced diet, and adverse effects of being on the diet are sometimes reported.

Spanish Adaptation of the Inventory Brief Child Abuse Potential and the Protective Factors Survey.

Child maltreatment is a public health problem with different consequences depending on the form of abuse. Measuring risk and protective factors has been a fertile ground for research, without involving instruments with sufficient evidence of validity. The aim of the study was to gather evidence of validity and reliability of the Inventory Brief Child Abuse Potential (IBCAP) and Protective Factors Survey (PFS) in the Mexican population. The instruments were translated into Spanish. In a non-probabilistic sample of 200 participants, the 7-factor model for the IBCAP [comparative fit index (CFI) = 0.984; root mean square error of approximation (RMSEA) = 0.067] and the 4-factor model for the PFS (CFI = 0.974; RMSEA = 0.061) were confirmed, showing adequate fit indices. Reliability was estimated and evidence of convergent, divergent, and discriminant validity was collected, controlling for effects of social desirability. We also report interpretability statistics of the scores. We achieved solid progress in the development of instrumentation that allows determining the presence or absence of protective and risk factors for child abuse.