Protocol to identify protein-protein interaction networks in Solanum tuberosum using transient TurboID-based proximity labeling.

Protein-protein interactions (PPIs) in crop plants remain largely unexplored. Here, we provide a protocol for identifying PPIs in potato (Solanum tuberosum) using TurboID-mediated proximity labeling. We transiently expressed constructs for a nucleus-located transcription factor and a plasma membrane-localized receptor-like kinase fused to TurboID to identify PPIs in potato leaves. We describe the plasmid construction, plant material, agroinfiltration, biotin treatment, protein isolation, free biotin removal, western blot analysis, and enrichment of biotinylated proteins for mass spectrometry analysis.

Effects of in utero exposure to fluoxetine on the gastrointestinal tract of rat offspring.

Perinatal exposure to selective serotonin reuptake inhibitors (SSRIs) has been shown to disrupt the development of serotonergic signaling pathways in the brain and enteric nervous system. Serotonin (5-hydroxytryptamine; 5-HT) signaling is critical for gastrointestinal homeostasis; changes in 5-HT expression and regulation have been associated with gastrointestinal diseases of motility and inflammation. We tested the hypothesis that perinatal exposure to the SSRI fluoxetine can influence the development of the gastrointestinal tract in exposed offspring. Female nulliparous Wistar rats were given fluoxetine (10 mg/kg) or vehicle control from 2 wk before mating until weaning; small and large intestines of female and male offspring were collected at postnatal days 1, 21 (P1, P21, respectively), and 6 mo of age. In histological preparations, the proportion of serotonergic neurons significantly increased in the colons of both female and male fluoxetine-exposed compared with control offspring at P21, a time point that signifies maximal exposure to fluoxetine. At 6 mo of age, male but not female fluoxetine-exposed offspring had a significant increase in circulating 5-HT, with a significant decrease in transcripts encoding the 5-HT2A receptor and monoamine oxidase as compared with control offspring. Measurement of spatiotemporal mapping of contractile activity of the small and large intestine at 6 mo of age revealed no changes in motility in the small bowel of fluoxetine-exposed offspring but revealed a significant increase in the frequency of colonic contractions in the female fluoxetine-exposed compared with control animals. Susceptibility to inflammation was examined at 6 mo using the dextran sulfate sodium model of acute colitis. In utero exposure to fluoxetine was not found to exacerbate colitis severity. These findings suggest that fluoxetine exposure during fetal and early postnatal development can lead to changes in serotonergic neurons at the peak of exposure with sex-specific changes in 5-HT signaling and colonic motility in adulthood.NEW & NOTEWORTHY There is increasing recognition of the relevance of in utero and early postnatal exposures in the developmental programming of the gastrointestinal tract. Perinatal exposure to selective serotonin reuptake inhibitors and antidepressant medications is of particular relevance as they are commonly prescribed during pregnancy, and serotonergic pathways play key roles during gastrointestinal development and in postnatal homeostasis. Here, we provide a comprehensive evaluation of clinically relevant outcomes of gastrointestinal motility and susceptibility to colitis in fluoxetine-exposed offspring and highlight changes in colonic serotonergic neurons at the peak of perinatal fluoxetine exposure with sex-dependent changes in serotonin signaling and colonic motility in adulthood.

Indoor air quality in a training centre used for sports practice.

Background: One of the measures for controlling the coronavirus disease 2019 (COVID-19) pandemic was the mass closure of gyms. This measure leads us to determine the differences between indoor and outdoor air quality. That is why the objective of this study was to analyse the indoor air quality of a sports centre catering to small groups and rehabilitation. Methods: The study was conducted in a single training centre, where 26 measurements were taken in two spaces (indoors and outdoors). The air quality index, temperature, relative humidity, total volatile compounds, carbon monoxide, ozone, formaldehyde, carbon dioxide, and particulate matter were measured indoors and outdoors using the same protocol and equipment. These measurements were taken twice, once in the morning and once in the afternoon, with all measurements made at the same time, 10 am and 6 pm, respectively. Additionally, four determinations of each variable were collected during each shift, and the number of people who had trained in the room and the number of trainers were counted. Results: In the different variables analysed, the results show that CO2 and RH levels are higher indoors than outdoors in both measurement shifts. Temperatures are higher outside than inside and, in the evening, than in the morning. TVOC, AQI and PM show less variation, although they are higher outdoors in the morning. CO is highest indoors. HCHO levels are almost negligible and do not vary significantly, except for a slight increase in the afternoon outside. Ozone levels are not significant. All the variables showed practically perfect reliability in all the measurements, except for ozone measured outside in the morning. On the other hand, the variables exhibit variations between indoors and outdoors during the morning and afternoon, except for the three types of PM. Also, the data show that all the main variables measured inside the sports training centre are similar between morning and afternoon. However, outside, temperature, relative humidity and HCHO levels show significant differences between morning and afternoon while no differences are observed for the other variables. Conclusion: The indoor air quality of the training centre assessed was good and met current regulations; some of its components even exhibited better levels than fresh air. This article is the first to measure indoor air quality in a sports training centre catering to rehabilitation and small groups.

The Clinical Utility of lncRNAs and Their Application as Molecular Biomarkers in Breast Cancer.

Given their tumor-specific and stage-specific gene expression, long non-coding RNAs (lncRNAs) have demonstrated to be potential molecular biomarkers for diagnosis, prognosis, and treatment response. Particularly, the lncRNAs DSCAM-AS1 and GATA3-AS1 serve as examples of this because of their high subtype-specific expression profile in luminal B-like breast cancer. This makes them candidates to use as molecular biomarkers in clinical practice. However, lncRNA studies in breast cancer are limited in sample size and are restricted to the determination of their biological function, which represents an obstacle for its inclusion as molecular biomarkers of clinical utility. Nevertheless, due to their expression specificity among diseases, such as cancer, and their stability in body fluids, lncRNAs are promising molecular biomarkers that could improve the reliability, sensitivity, and specificity of molecular techniques used in clinical diagnosis. The development of lncRNA-based diagnostics and lncRNA-based therapeutics will be useful in routine medical practice to improve patient clinical management and quality of life.

Transformative learning for a sustainable and healthy future through ecosystem approaches to health: insights from 15 years of co-designed ecohealth teaching and learning experiences.

This paper presents insights from the work of the Canadian Community of Practice in Ecosystem Approaches to Health (CoPEH-Canada) and 15 years (2008-2022) of land-based, transdisciplinary, learner-centred, transformative learning and training. We have oriented our learning approaches to Head, Hands, and Heart, which symbolise cognitive, psychomotor, and affective learning, respectively. Psychomotor and affective learning are necessary to grapple with and enact far-reaching structural changes (eg, decolonisation) needed to rekindle healthier, reciprocal relationships with nature and each other. We acknowledge that these approaches have been long understood by Indigenous colleagues and communities. We have developed a suite of teaching techniques and resources through an iterative and evolving pedagogy based on participatory approaches and operating reciprocal, research-pedagogical cycles; integrated different approaches and ways of knowing into our pedagogy; and built a networked Community of Practice for continued learning. Planetary health has become a dominant framing for health-ecosystem interactions. This Viewpoint underscores the depth of existing scholarship, collaboration, and pedagogical expertise in ecohealth teaching and learning that can inform planetary health education approaches.

Efectos de la quiebra de la conducta sedentaria sobre parámetros fisiológicos y cognitivos en pediatría

Introducción: La actividad física resulta relevante para la salud física y mental. La conducta sedentaria diaria afecta la salud, y realizar actividad física contribuye al mantenimiento de la salud mental. Objetivo: Analizar, en población pediátrica, la evidencia científica actualizada relacionada con los efectos de las quiebras de la conducta sedentaria sobre marcadores cardiometabólicos y la función cognitiva. Métodos: Revisión sistemática de estudios publicados en bases de datos científicas: PubMed, Cochrane, Science Direct, Medline. Los límites de la búsqueda se ubicaron en estudios publicados entre 2016 y 2022, realizados en niños y niñas entre 4 y 13 años. Análisis y síntesis de la información: Se identificaron 127 artículos con 2522 pacientes. Se incluyeron en el análisis final 10 artículos y se consideraron dos subgrupos; a) pacientes con quiebres de la conducta sedentaria y su efecto sobre marcadores cardiometabólicos y b) pacientes con quiebras de la conducta sedentaria y su efecto sobre funciones cognitivas. Conclusiones: Los resultados obtenidos indicaron que las intervenciones con protocolos de quiebra de la conducta sedentaria constituyen un método eficaz para mejorar indicadores de salud cardiometabólica e indicadores de funciones cognitivas en la población pediátrica.

Introduction: Physical activity is relevant to physical and mental health. Daily sedentary behavior affects health, and physical activity contributes to the maintenance of mental health. Objective: To analyze, in a pediatric population, the updated scientific evidence related to the effects of sedentary behavioral breaks on cardiometabolic markers and cognitive function. Methods: Systematic review of studies published in scientific databases like: PubMed, Cochrane, Science Direct, Medline. The search limits were placed on studies published between 2016 and 2022, and conducted in boys and girls aged 4 to 13 years. Analysis and synthesis of information: 127 articles with 2522 patients were identified. Ten articles were included in the final analysis and two subgroups were considered: a) patients with breaks in sedentary behavior and their effect on cardiometabolic markers and b) patients with breaks in sedentary behavior and their effect on cognitive functions. Conclusions: The results obtained indicated that interventions with sedentary behavior breakdown protocols constitute an effective method to improve cardiometabolic health markers and cognitive function markers in the pediatric population.

EDS1 complexes are not required for PRR responses and execute TNL-ETI from the nucleus in Nicotiana benthamiana.

Heterodimeric complexes incorporating the lipase-like proteins EDS1 with PAD4 or SAG101 are central hubs in plant innate immunity. EDS1 functions encompass signal relay from TIR domain-containing intracellular NLR-type immune receptors (TNLs) towards RPW8-type helper NLRs (RNLs) and, in Arabidopsis thaliana, bolstering of signaling and resistance mediated by cell-surface pattern recognition receptors (PRRs). Increasing evidence points to the activation of EDS1 complexes by small molecule binding. We used CRISPR/Cas-generated mutant lines and agroinfiltration-based complementation assays to interrogate functions of EDS1 complexes in Nicotiana benthamiana. We did not detect impaired PRR signaling in N. benthamiana lines deficient in EDS1 complexes or RNLs. Intriguingly, in assays monitoring functions of SlEDS1-NbEDS1 complexes in N. benthamiana, mutations within the SlEDS1 catalytic triad could abolish or enhance TNL immunity. Furthermore, nuclear EDS1 accumulation was sufficient for N. benthamiana TNL (Roq1) immunity. Reinforcing PRR signaling in Arabidopsis might be a derived function of the TNL/EDS1 immune sector. Although Solanaceae EDS1 functionally depends on catalytic triad residues in some contexts, our data do not support binding of a TNL-derived small molecule in the triad environment. Whether and how nuclear EDS1 activity connects to membrane pore-forming RNLs remains unknown.

Comprehensive and systematic characterization of multi-functionalized cisplatin nano-conjugate: from the chemistry and proteomic biocompatibility to the animal model.

BACKGROUND: Nowadays, nanoparticles (NPs) have evolved as multifunctional systems combining different custom anchorages which opens a wide range of applications in biomedical research. Thus, their pharmacological involvements require more comprehensive analysis and novel nanodrugs should be characterized by both chemically and biological point of view. Within the wide variety of biocompatible nanosystems, iron oxide nanoparticles (IONPs) present mostly of the required features which make them suitable for multifunctional NPs with many biopharmaceutical applications. RESULTS: Cisplatin-IONPs and different functionalization stages have been broadly evaluated. The potential application of these nanodrugs in onco-therapies has been assessed by studying in vitro biocompatibility (interactions with environment) by proteomics characterization the determination of protein corona in different proximal fluids (human plasma, rabbit plasma and fetal bovine serum),. Moreover, protein labeling and LC-MS/MS analysis provided more than 4000 proteins de novo synthetized as consequence of the nanodrugs presence defending cell signaling in different tumor cell types (data available via ProteomeXchanges with identified PXD026615). Further in vivo studies have provided a more integrative view of the biopharmaceutical perspectives of IONPs. CONCLUSIONS: Pharmacological proteomic profile different behavior between species and different affinity of protein coating layers (soft and hard corona). Also, intracellular signaling exposed differences between tumor cell lines studied. First approaches in animal model reveal the potential of theses NPs as drug delivery vehicles and confirm cisplatin compounds as strengthened antitumoral agents.

Prenatal nicotine exposure leads to decreased histone H3 lysine 9 (H3K9) methylation and increased p66shc expression in the neonatal pancreas.

Prenatal exposure to nicotine, tobacco's major addictive constituent, has been shown to reduce birth weight and increases apoptosis, oxidative stress, and mitochondrial dysfunction in the postnatal pancreas. Given that upregulated levels of the pro-oxidative adapter protein p66shc is observed in growth-restricted offspring and is linked to beta-cell apoptosis, the goal of this study was to investigate whether alterations in p66shc expression underlie the pancreatic deficits in nicotine-exposed offspring. Maternal administration of nicotine in rats increased p66shc expression in the neonatal pancreas. Similarly, nicotine treatment augmented p66shc expression in INS-1E pancreatic beta cells. Increased p66shc expression was also associated with decreased histone H3 lysine 9 methylation. Finally, nicotine increased the expression of Kdm4c, a key histone lysine demethylase, and decreased Suv39h1, a critical histone lysine methyltransferase. Collectively, these results suggest that upregulation of p66shc through posttranslational histone modifications may underlie the reported adverse outcomes of nicotine exposure on pancreatic function.

Trends in breast-conserving surgery in Mexico after the implementation of a public health insurance system.

Although breast conserving surgery is the standard of care for patients with localized breast cancer in high-income countries, little is known about its use in developing countries, where disparities in access to treatment may lead to an increased use of mastectomy. We examined the use of breast conserving surgery at a Mexican cancer center after the implementation of a public insurance program aimed at providing coverage for previously uninsured patients. Between 2006 and 2016, 4519 women received surgical treatment for breast cancer, of which 39% had early-stage disease. The proportion of patients treated with breast conserving surgery increased from 10% in the 2006-2009 period to 33% in the 2013-2016 period, with most of this increase occurring among women with early-stage disease (17-52%). Improving access to care and reducing the financial burden of breast cancer in developing countries may lead to an increased use of breast conserving surgery.

Red light imaging for programmed cell death visualization and quantification in plant-pathogen interactions.

Studies on plant-pathogen interactions often involve monitoring disease symptoms or responses of the host plant to pathogen-derived immunogenic patterns, either visually or by staining the plant tissue. Both these methods have limitations with respect to resolution, reproducibility, and the ability to quantify the results. In this study we show that red light detection by the red fluorescent protein (RFP) channel of a multipurpose fluorescence imaging system that is probably available in many laboratories can be used to visualize plant tissue undergoing cell death. Red light emission is the result of chlorophyll fluorescence on thylakoid membrane disassembly during the development of a programmed cell death process. The activation of programmed cell death can occur during either a hypersensitive response to a biotrophic pathogen or an apoptotic cell death triggered by a necrotrophic pathogen. Quantifying the intensity of the red light signal enables the magnitude of programmed cell death to be evaluated and provides a readout of the plant immune response in a faster, safer, and nondestructive manner when compared to previously developed chemical staining methodologies. This application can be implemented to screen for differences in symptom severity in plant-pathogen interactions, and to visualize and quantify in a more sensitive and objective manner the intensity of the plant response on perception of a given immunological pattern. We illustrate the utility and versatility of the method using diverse immunogenic patterns and pathogens.

Effects of dibenzothiophene, a sulfur-containing heterocyclic aromatic hydrocarbon, and its alkylated congener, 2,4,7-trimethyldibenzothiophene, on placental trophoblast cell function.

Worldwide demand for petroleum products has resulted in increased oil and gas activities in many countries. Conventional and unconventional oil and gas extraction, production, and transport lead to increased levels of petroleum-derived polycyclic aromatic hydrocarbons (PAHs) in the environment. PAH exposure has profound effects on reproduction by affecting pathways involved in placental trophoblast cell function and impairing normal placental development and function-key contributors to reproductive success. However, other components found in petroleum and wastewaters from oil and gas extraction, including the sulfur-containing heterocyclic aromatic compounds such as dibenzothiophene (DBT) and its alkylated derivatives, may also impact reproductive success. The goal of this study was to examine the effect of exposure to DBT, a compound commonly detected in the environment, and one of its alkylated analogues, 2,4,7-trimethyldibenzothiophene (2,4,7-DBT), on steroidogenic and angiogenic pathways critical for mammalian development in placental trophoblast cells (HTR-8/SVneo cells). 2,4,7-DBT but not DBT increased estradiol output in association with increased tube-like formation (surrogate for angiogenesis). These changes in angiogenesis did not appear to be related to altered expression of the key placental angiogenic gene targets (ANGPTL4, VEGFA, and PGF). Neither compound showed a concentration related effect on progesterone synthesis or its receptor expression. Our results suggest that 2,4,7-DBT can disrupt key pathways important for placental trophoblast function and highlight the importance of determining the impact of exposure to both parent and alkylated compounds. Further, these data suggest that exposure to sulfur-containing heterocyclic aromatic compounds may lead to placental dysfunction and impact reproductive success at environmentally relevant levels.

The effects of a technical mixture of naphthenic acids on placental trophoblast cell function.

There is considerable concern that naphthenic acids (NA) related to oil extraction can negatively impact reproduction in mammals, yet the mechanisms are unknown. Since placental dysfunction is central to many adverse pregnancy outcomes, the goal of this study was to determine the effects of NA exposure on placental trophoblast cell function. HTR-8/SVneo cells were exposed to a commercial technical NA mixture for 24 hours to assess transcriptional regulation of placentation-related pathways and functional assessment of migration, invasion, and angiogenesis. Pathway analysis suggests that NA treatment resulted in increased epithelial-to-mesenchymal transition. However, there was reduced migration and invasive potential. NA treatment increased angiogenesis-related pathways with a concomitant increase in tube formation. Since decreased trophoblast invasion/migration and aberrant angiogenesis have been associated with placental dysfunction, these findings suggest that it is biologically plausible that exposure to NA may result in altered placental development and/or function.

Test cardiopulmonar: una herramienta de utilidad diagnóstica y pronóstica / Cardiopulmonary exercise test for diagnostic and prognostic purposes

The functional assessment of patients with dyspnea usually uses static or submaximal exercise tests, which provide limited information because they do not expose patients to the real situation that causes exercise intolerance. The cardiopulmonary exercise test (CPET) is an increasingly used tool that can be used in these circumstances. It determines peak oxygen consumption, anaerobic threshold and cardiac and respiratory reserves, measuring oxygen uptake and carbon dioxide production during standardized exercise conditions. It is useful for risk assessment in cardiothoracic surgery and can provide valuable information such as the timing for transplant in patients with severe chronic disease. The test is non-invasive, has a short duration, and exhibits an adequate safety profile in specialized centers. It is mainly indicated for the dynamic evaluation of athletes or patients with heart, respiratory, and neuromuscular diseases, it is essential part of the study of dyspnea of unknown origin, and in the prognostic assessment of patients who face highly complex interventions. This review provides a comprehensive review of CPET with emphasis on its main indications in healthy people, athletes and, in particular, in functional evaluation of patients with exercise limitations in the context of their chronic diseases.

Spontaneous perforation of the common bile duct in a pediatric patient. Case report and short review of the literature.

INTRODUCTION: Spontaneous perforation of the common bile duct is rare. The cause is usually unknown, although it is sometimes related to the malformation of the bile duct. CLINICAL CASE: Female of 1 year, with abdominal distention, hyporexia and fever, tomography with ascites, surgical findings included retroperitoneal bilioma, peritonitis and perforation of common bile duct. Block resection and primary hepato-duodenal anastomosis were performed. DISCUSSION: Spontaneous perforation of the common bile duct tend to evolve insidiously and delay in diagnosis is frequent. Surgical management is decisive, and there are different techniques described. CONCLUSION: A high index of suspicion is important, treatment must be individualized according to each patient.

INTRODUCCIÓN: La perforación espontánea del colédoco es rara. La causa generalmente se desconoce, aunque en ocasiones guarda relación con una malformación de la vía biliar. CASO CLÍNICO: Niña de 1 año, con distensión abdominal, hiporexia y fiebre, tomografía con ascitis, se interviene y se encuentra bilioma retroperitoneal, peritonitis y perforación de colédoco. Se realiza resección en bloque y anastomosis hepatoduodenal primaria. DISCUSIÓN: La perforación espontánea del colédoco tiende a evolucionar insidiosamente y el retraso en el diagnóstico es frecuente. El manejo quirúrgico es resolutivo; existen distintas técnicas descritas. CONCLUSIÓN: Es importante un alto índice de sospecha. El tratamiento se debe individualizar en cada paciente.

Adverse effects of naphthenic acids on reproductive health: A focus on placental trophoblast cells.

There is considerable concern that naphthenic acids (NA) related to oil extraction can negatively impact reproduction in mammals yet the mechanisms are unknown. Since placental dysfunction is central to many adverse pregnancy outcomes, the goal of this study was to determine the effects of NA exposure on placental trophoblast cell function. Htr-8/SVneo cells were exposed to a commercial technical NA mixture (Sigma-Aldrich) for 24 h to assess steroid production, markers of inflammation and oxidative stress. NA treatment significantly altered steroid production; progesterone was decreased at all doses tested, whereas there was a significant increase in testosterone production (125 mg/L only). There were no effects on estradiol production. In addition, NA treatment resulted in increased markers of inflammation (interleukin 1ß and prostaglandin E2) and oxidative damage to lipids and nucleic acids. These findings suggest that it is biologically plausible that NA exposure may contribute to placental dysfunction.

Δ9-Tetrahydrocannabinol leads to endoplasmic reticulum stress and mitochondrial dysfunction in human BeWo trophoblasts.

While studies have demonstrated that the main psychoactive component of cannabis, Δ9-tetrahydrocannabinol (Δ9-THC) alone induces placental insufficiency and fetal growth restriction, the underlying mechanisms remain elusive. Given that both (i) endoplasmic reticulum (ER) stress in pregnancy and (ii) gestational exposure to Δ9-THC leads to placental deficiency, we hypothesized that Δ9-THC may directly induce placental ER stress, influencing trophoblast gene expression and mitochondrial function. BeWo human trophoblast cells treated with Δ9-THC (3-30 µM) led to a dose-dependent increase in all ER stress markers and CHOP; these effects could be blocked with CB1R/CB2R antagonists. Moreover, expression of ER stress-sensitive genes ERRγ, VEGFA, and FLT-1 were increased by Δ9-THC, and abrogated with the ER stress inhibitor TUDCA. Δ9-THC also diminished mitochondrial respiration and ATP-coupling due to decreased abundance of mitochondrial chain complex proteins. Collectively, these findings indicate that Δ9-THC can directly augment ER stress resulting in aberrant placental gene expression and impaired mitochondrial function.

Synthesis of α-methylstilbenes using an aqueous Wittig methodology and application toward the development of potent human aromatase inhibitors.

The development of aqueous Wittig methodology for the synthesis of α-methylstilbenes using tripropylphosphine-derived phosphonium salts is described. The Wittig olefination reaction was high yielding and allowed isolation of stilbenes by simple filtration and washing with water. The novel phosphonium salts employed were accessed via a highly efficient, regioselective addition of hydrogen bromide to styrenes. Application of the α-methylstilbenes toward the synthesis of a collection of stilbenoid-triazoles is reported and their inhibition of CYP450 19A1 (aromatase) investigated. The overall structure-activity profile provided additional evidence on the aryl halide-ketone bioisostere hypothesis and identified 6c as a potent inhibitor of aromatase in vitro (Ki = 8 nM).