The stress experienced in an emergency medical service (EMS): A descriptive study.

INTRODUCTION: Stress, described as an adaptation of the body to an event, is a considerable problem among health workers, especially for those who work in emergency situations, as they very often have to face complex situations. It has been proven that stress affects the performance of health professionals, which is why it is interesting to measure it in these situations, to be able to know what methods to implement to reduce it in future events. Despite having previous measurements in healthcare personnel during clinical simulations, this study is relevant because stress has never before been measured in EMS professionals when performing their work. OBJECTIVE: To determine the acute stress experienced by professionals in an Emergency Medical Service (EMS) when handling five types of clinical emergencies. As secondary objectives, to determine if there were differences in the increases in stress in relation to sex, age, profession, team, and above all, type of emergency handled. METHOD: The following were measured for the analysis: physiological (mean heart rate (HR), systolic (SBP) and diastolic (DBP) blood pressure), and biochemical (salivary alpha amylase (sAA) activity) markers, before (Basal) and after (Post-Stress) the interventions of 27 professionals from an EMS. RESULTS: In general, the results obtained showed significant differences between the baseline measurement of physiological and biochemical markers versus the post-exposure measurement. Some of the differences in stress levels in relation to sex and professional role are striking. CONCLUSION: The determination of acute stress experienced by professionals from an EMS in a real emergency showed significant increases in the sAA enzymatic marker of acute stress. These results are the first data published in this regard, and could be used as a reference to follow in clinical simulation in the training of students and the training of nursing professionals. IMPLICATIONS FOR CLINICAL PRACTICE: Evidence based studies are needed to improve the education and training of emergency and intensive care professionals. The results from are a great step in the analysis of the real stress that professionals are subjected to when they handle different emergencies.

Safety of assisted reproductive techniques in gynecological cancer patients.

PURPOSE OF REVIEW: Gynecological cancer is a very important cause of comorbidity and mortality in women. The current delay in motherhood is increasing the incidence of women under 40 years of age that have not yet achieved their maternity goals when they are diagnosed and standard treatment negatively impacts the reproductive potential of cancer survivors. In this review, we update the information available about the safety of fertility-sparing treatments in young gynecological cancer patients, as well as the safety and efficacy of assisted reproductive techniques (ART) in such group. We also evaluate the long-term gynecological cancer risk in women requiring ART. RECENT FINDINGS: Although eligibility criteria continue to be very strict, there are more and more reports of fertility-sparing approaches outside of what traditionally has been considered safe. Molecular assessment is starting to be used in the selection of appropriate candidates. Data increasingly shows the long term safety and the efficacy of ART and pregnancy in these patients. SUMMARY: Appropriate selection is key to safely preconize fertility-sparing alternatives. Because subfertility may be a result of these procedures, ART could be indicated in this setting. Neither ART nor pregnancy appear to increase recurrences or affect survival rates.

Nucleus-translocated mitochondrial cytochrome c liberates nucleophosmin-sequestered ARF tumor suppressor by changing nucleolar liquid-liquid phase separation.

The regular functioning of the nucleolus and nucleus-mitochondria crosstalk are considered unrelated processes, yet cytochrome c (Cc) migrates to the nucleus and even the nucleolus under stress conditions. Nucleolar liquid-liquid phase separation usually serves the cell as a fast, smart mechanism to control the spatial localization and trafficking of nuclear proteins. Actually, the alternative reading frame (ARF), a tumor suppressor protein sequestered by nucleophosmin (NPM) in the nucleoli, is shifted out from NPM upon DNA damage. DNA damage also triggers early translocation of respiratory Cc to nucleus before cytoplasmic caspase activation. Here, we show that Cc can bind to nucleolar NPM by triggering an extended-to-compact conformational change, driving ARF release. Such a NPM-Cc nucleolar interaction can be extended to a general mechanism for DNA damage in which the lysine-rich regions of Cc-rather than the canonical, arginine-rich stretches of membrane-less organelle components-controls the trafficking and availability of nucleolar proteins.

Deciphering the therapeutical potentials of rosmarinic acid.

Lemon balm is herbal tea used for soothing stomach cramps, indigestion, and nausea. Rosmarinic acid (RA) is one of its chemical constituents known for its therapeutic potentials against cancer, inflammatory and neuronal diseases such as the treatment of neurofibromatosis or prevention from Alzheimer's diseases (AD). Despite efforts, recovery and purification of RA in high yields has not been entirely successful. Here, we report its aqueous extraction with optimal conditions and decipher the structure by nuclear magnetic resonance (NMR) spectroscopy. Using various physical-chemical and biological assays, we highlight its anti-aggregation inhibition potentials against the formation of Tau filaments, one of the hallmarks of AD. We then examine its anti-cancer potentials through reduction of the mitochondrial reductase activity in tumor cells and investigate its electrochemical properties by cyclic voltammetry. Our data demonstrates that RA is a prominent biologically active natural product with therapeutic potentials for drug discovery in AD, cancer therapy and inflammatory diseases.

Peptide Conjugates Derived from flg15, Pep13, and PIP1 That Are Active against Plant-Pathogenic Bacteria and Trigger Plant Defense Responses.

Thirty peptide conjugates were designed by combining an antimicrobial peptide (BP16, BP100, BP143, KSL-W, BP387, or BP475) at the N- or C-terminus of a plant defense elicitor peptide (flg15, BP13, Pep13, or PIP1). These conjugates were highly active in vitro against six plant-pathogenic bacteria, especially against Xanthomonas arboricola pv. pruni, Xanthomonas fragariae and Xanthomonas axonopodis pv. vesicatoria. The most active peptides were those incorporating Pep13. The order of the conjugation influenced the antibacterial activity and the hemolysis. Regarding the former, peptide conjugates incorporating the elicitor peptide flg15 or Pep13 at the C-terminus were, in general, more active against Pseudomonas syringae pv. actinidiae and P. syringae pv. syringae, whereas those bearing these elicitor peptides at the N-terminus displayed higher activity against Erwinia. amylovora and the Xanthomonas species. The best peptide conjugates displayed MIC values between 0.8 and 12.5 µM against all the bacteria tested and also had low levels of hemolysis and low phytotoxicity. Analysis of the structural and physicochemical parameters revealed that a positive charge ranging from +5 to +7 and a moderate hydrophobic moment/amphipathic character is required for an optimal biological profile. Interestingly, flg15-BP475 exhibited a dual activity, causing the upregulation of the same genes as flg15 and reducing the severity of bacterial spot in tomato plants with a similar or even higher efficacy than copper oxychloride. Characterization by nuclear magnetic resonance (NMR) of the secondary structure of flg15-BP475 showed that residues 10 to 25 fold into an α-helix. This study establishes trends to design new bifunctional peptides useful against plant diseases caused by plant-pathogenic bacteria. IMPORTANCE The consequences of plant pathogens on crop production together with the lack of effective and environmentally friendly pesticides evidence the need of new agents to control plant diseases. Antimicrobial and plant defense elicitor peptides have emerged as good candidates to tackle this problem. This study focused on combining these two types of peptides into a single conjugate with the aim to potentiate the activity of the individual fragments. Differences in the biological activity of the resulting peptide conjugates were obtained depending on their charge, amphipathicity, and hydrophobicity, as well as on the order of the conjugation of the monomers. This work provided bifunctional peptide conjugates able to inhibit several plant-pathogenic bacteria, to stimulate plant defense responses, and to reduce the severity of bacterial spot in tomato plants. Thus, this study could serve as the basis for the development of new antibacterial/plant defense elicitor peptides to control bacterial plant pathogens.

Midkine Interaction with Chondroitin Sulfate Model Synthetic Tetrasaccharides and Their Mimetics: The Role of Aromatic Interactions.

Midkine (MK) is a neurotrophic factor that participates in the embryonic central nervous system (CNS) development and neural stem cell regulation, interacting with sulfated glycosaminoglycans (GAGs). Chondroitin sulfate (CS) is the natural ligand in the CNS. In this work, we describe the interactions between a library of synthetic models of CS-types and mimics. We did a structural study of this library by NMR and MD (Molecular Dynamics), concluding that the basic shape is controlled by similar geometry of the glycosidic linkages. Their 3D structures are a helix with four residues per turn, almost linear. We have studied the tetrasaccharide-midkine complexes by ligand observed NMR techniques and concluded that the shape of the ligands does not change upon binding. The ligand orientation into the complex is very variable. It is placed inside the central cavity of MK formed by the two structured beta-sheets domains linked by an intrinsically disordered region (IDR). Docking analysis confirmed the participation of aromatics residues from MK completed with electrostatic interactions. Finally, we test the biological activity by increasing the MK expression using CS tetrasaccharides and their capacity in enhancing the growth stimulation effect of MK in NIH3T3 cells.

D-Amino Acid-Containing Lipopeptides Derived from the Lead Peptide BP100 with Activity against Plant Pathogens.

From a previous collection of lipopeptides derived from BP100, we selected 18 sequences in order to improve their biological profile. In particular, analogues containing a D-amino acid at position 4 were designed, prepared, and tested against plant pathogenic bacteria and fungi. The biological activity of these sequences was compared with that of the corresponding parent lipopeptides with all L-amino acids. In addition, the influence of the length of the hydrophobic chain on the biological activity was evaluated. Interestingly, the incorporation of a D-amino acid into lipopeptides bearing a butanoyl or a hexanoyl chain led to less hemolytic sequences and, in general, that were as active or more active than the corresponding all L-lipopeptides. The best lipopeptides were BP475 and BP485, both incorporating a D-Phe at position 4 and a butanoyl group, with MIC values between 0.8 and 6.2 µM, low hemolysis (0 and 24% at 250 µM, respectively), and low phytotoxicity. Characterization by NMR of the secondary structure of BP475 revealed that the D-Phe at position 4 disrupts the α-helix and that residues 6 to 10 are able to fold in an α-helix. This secondary structure would be responsible for the high antimicrobial activity and low hemolysis of this lipopeptide.

Virtual reality in cardiopulmonary resuscitation training: a randomized trial. / La realidad virtual como método de enseñanza de la reanimación cardiopulmonar: un estudio aleatorizado.

OBJECTIVES: To assess the efficacy of virtual reality (VR) in cardiopulmonary resuscitation (CPR) training. MATERIAL AND METHODS: Experimental, analytic, cross-sectional study of a CPR training method using VR. Participants were randomly assigned to train in a control group or a VR group. RESULTS: The mean (SD) scores on a scale of 10 after training were 9.28 (0.91) in the VR group and 7.78 (1.63) in the control group, for a mean difference of 1.49 (95% CI, 0.96-2.02; P<.001). The VR group achieved a mean of 97.5 (9.7) compressions/min, versus 80.9 (7.7) compressions/min in the control group, for a mean difference of 16.6 compressions/min (95% CI, 15.0-18.2; P=.003). The mean compression depth in the VR group was 34.0 (6.5) mm, versus 27.9 (4.9) mm in the control group, for a mean difference of 6.7 (95% CI, 5.7-7.8; P<.001). CONCLUSION: Training with VR can improve CPR theoretical knowledge and practical skills.

OBJETIVO: Analizar la eficacia de la realidad virtual (RV) en la formación en reanimación cardiopulmonar (RCP). METODO: Estudio experimental, analítico, transversal para analizar el aprendizaje en RCP a través de la RV, en el que los participantes fueron asignados aleatoriamente en grupo control (GC) y grupo RV (GRV). RESULTADOS: La nota del test fue de GRV fue 9,28 (DE 0,91) y el de GC 7,78 (DE 1,63) [diferencia de medias 1,49 (IC95% 0,96-2,02), p < 0,001]. El ritmo medio de las compresiones fue 97,5 (DE 9,7) compresiones/min para el GRV y 80,9 (DE 7,7) compresiones/min para el GC [diferencia de medias 16,6 (IC95% 15,0-18,2), p = 0,003]. La profundidad media fue 34,0 (DE 6,5) mm para el GRV y 27,3 (DE 4,9) mm para el GC [diferencia de medias 6,7 (IC95% 5,7- 7,8), p < 0,001]. CONCLUSIONES: La RV es un método de enseñanza de RCP capaz de mejorar los conocimientos teóricos y habilidades prácticas.

Cytochrome c speeds up caspase cascade activation by blocking 14-3-3ε-dependent Apaf-1 inhibition.

Apoptosis is a highly regulated form of programmed cell death, essential to the development and homeostasis of multicellular organisms. Cytochrome c is a central figure in the activation of the apoptotic intrinsic pathway, thereby activating the caspase cascade through its interaction with Apaf-1. Our recent studies have revealed 14-3-3ε (a direct inhibitor of Apaf-1) as a cytosolic cytochrome c target. Here we explore the cytochrome c / 14-3-3ε interaction and show the ability of cytochrome c to block 14-3-3ε-mediated Apaf-1 inhibition, thereby unveiling a novel function for cytochrome c as an indirect activator of caspase-9/3. We have used calorimetry, NMR spectroscopy, site mutagenesis and computational calculations to provide an insight into the structural features of the cytochrome c / 14-3-3ε complex. Overall, these findings suggest an additional cytochrome c-mediated mechanism to modulate apoptosome formation, shedding light onto the rigorous apoptotic regulation network.

Learning cardiopulmonary resuscitation theory with face-to-face versus audiovisual instruction for secondary school students: a randomized controlled trial. / Ensayo clínico aleatorizado controlado que compara la formación presencial frente a la no presencial en el aprendizaje teórico de la reanimación cardiopulmonar entre los estudiantes de secundaria.

OBJECTIVES: To compare secondary students' learning of basic life support (BLS) theory and the use of an automatic external defibrillator (AED) through face-to-face classroom instruction versus educational video instruction. MATERIAL AND METHODS: A total of 2225 secondary students from 15 schools were randomly assigned to one of the following 5 instructional groups: 1) face-to-face instruction with no audiovisual support, 2) face-to-face instruction with audiovisual support, 3) audiovisual instruction without face-to-face instruction, 4) audiovisual instruction with face-to-face instruction, and 5) a control group that received no instruction. The students took a test of BLS and AED theory before instruction, immediately after instruction, and 2 months later. RESULTS: The median (interquartile range) scores overall were 2.33 (2.17) at baseline, 5.33 (4.66) immediately after instruction (P<.001) and 6.00 (3.33) (P<.001). All groups except the control group improved their scores. Scores immediately after instruction and 2 months later were statistically similar after all types of instruction. CONCLUSION: No significant differences between face-to-face instruction and audiovisual instruction for learning BLS and AED theory were found in secondary school students either immediately after instruction or 2 months later.

OBJETIVO: Comparar la formación presencial, mediante una clase teórica, frente a la formación no presencial, con un método audiovisual con y sin refuerzo posterior, en el aprendizaje teórico del soporte vital básico (SVB) y el desfibrilador externo automático (DEA) entre los estudiantes de secundaria. METODO: Se llevó a cabo un ensayo clínico aleatorizado que incluyó a 2.225 estudiantes de secundaria procedentes de 15 centros educativos que fueron asignados al azar a uno de los siguientes cinco grupos: 1) Grupo formación presencial sin refuerzo; 2) Grupo formación presencial con refuerzo; 3) Grupo formación audiovisual sin refuerzo; 4) Grupo formación audiovisual con refuerzo; 5) Grupo control. Se realizó un test sobre aspectos teóricos del SVB y DEA antes, después y a los 2 meses de la estrategia formativa. RESULTADOS: Los resultados mostraron diferencias estadísticamente significativas en todos los grupos, excepto el grupo control, entre la puntuación obtenida en el test basal 2,33 (RIC 2,17) y el test inmediato 5,33 (RIC 4,66) (p < 0,001), y entre el test basal y el test final 6,00 (RIC 3,33) (p < 0,001). No hubo diferencias en el aprendizaje inmediato y a los 2 meses entre los diferentes tipos de formaciones. CONCLUSIONES: No se encontraron diferencias entre la formación presencial mediante charlas teóricas y la formación no presencial con método audiovisual en el aprendizaje teórico inmediato y a los dos meses en el aprendizaje teórico del SVB y el DEA entre los estudiantes de secundaria.

Bone Marrow-Derived Cell Recruitment to the Neurosensory Retina and Retinal Pigment Epithelial Cell Layer Following Subthreshold Retinal Phototherapy.

Purpose: We investigated whether subthreshold retinal phototherapy (SRPT) was associated with recruitment of bone marrow (BM)-derived cells to the neurosensory retina (NSR) and RPE layer. Methods: GFP chimeric mice and wild-type (WT) mice were subjected to SRPT using a slit-lamp infrared laser. Duty cycles of 5%, 10%, 15%, and 20% (0.1 seconds, 250 mW, spot size 50 µm) with 30 applications were placed 50 to 100 µm from the optic disc. In adoptive transfer studies, GFP+ cells were given intravenously immediately after WT mice received SRPT. Immunohistochemistry was done for ionized calcium-binding adapter molecule-1 (IBA-1+), CD45, Griffonia simplicifolia lectin isolectin B4, GFP or cytokeratin). Expression of Ccl2, Il1b, Il6, Hspa1a, Hsp90aa1, Cryab, Hif1a, Cxcl12, and Cxcr4 mRNA and flow cytometry of the NSR and RPE-choroid were performed. Results: Within 12 to 24 hours of SRPT, monocytes were detected in the NSR and RPE-choroid. Detection of reparative progenitors in the RPE occurred at 2 weeks using flow cytometry. Recruitment of GFP+ cells to the RPE layer occurred in a duty cycle-dependent manner in chimeric mice and in mice undergoing adoptive transfer. Hspa1a, Hsp90aa1, and Cryab mRNAs increased in the NSR at 2 hours post laser; Hif1a, Cxcl12, Hspa1a increased at 4 hours in the RPE-choroid; and Ccl2, Il1b, Ifng, and Il6 increased at 12 to 24 hours in the RPE-choroid. Conclusions: SRPT induces monocyte recruitment to the RPE followed by hematopoietic progenitor cell homing at 2 weeks. Recruitment occurs in a duty cycle-dependent manner and potentially could contribute to the therapeutic efficacy of SRPT.

Hematopoietic stem/progenitor involvement in retinal microvascular repair during diabetes: Implications for bone marrow rejuvenation.

The widespread nature of diabetes affects all organ systems of an individual including the bone marrow. Long-term damage to the cellular and extracellular components of the bone marrow leads to a rapid decline in the bone marrow-hematopoietic stem/progenitor cells (HS/PCs) compartment. This review will highlight the importance of bone marrow microenvironment in maintaining bone marrow HS/PC populations and the contribution of these key populations in microvascular repair during the natural history of diabetes. The autonomic nervous system can initiate and propagate bone marrow dysfunction in diabetes. Systemic pharmacological strategies designed to protect the bone marrow-HS/PC population from diabetes induced-oxidative stress and advanced glycation end product accumulation represent a new approach to target diabetic retinopathy progression. Protecting HS/PCs ensures their participation in vascular repair and reduces the risk of vasogdegeneration occurring in the retina.

Glycodendrimers as Chondroitin Sulfate Mimetics: Synthesis and Binding to Growth Factor Midkine.

Chondroitin sulfate (CS) is a member of the glycosaminoglycan (GAG) family, a class of polysaccharides implicated in relevant biological functions. The structural complexity of these carbohydrates demands the development of simple glycomimetics as useful tools to study the biological processes in which GAGs are involved. In this work we described the synthesis of the disaccharide unit of the CS-E (GlcA-GalNAc(4,6-di-OSO3 )), in a multivalent presentation. Using a fluorescence polarization competition assay we have demonstrated that a hexavalent dendrimer of this disaccharide interact with midkine, in the low micromolar range. This result highlights the potency of these disaccharide-displaying multivalent systems as interesting mimetics of longer and synthetically more complex GAG oligosaccharides.

Interactions between a Heparin Trisaccharide Library and FGF-1 Analyzed by NMR Methods.

FGF-1 is a potent mitogen that, by interacting simultaneously with Heparan Sulfate Glycosaminoglycan HSGAG and the extracellular domains of its membrane receptor (FGFR), generates an intracellular signal that finally leads to cell division. The overall structure of the ternary complex Heparin:FGF-1:FGFR has been finally elucidated after some controversy and the interactions within the ternary complex have been deeply described. However, since the structure of the ternary complex was described, not much attention has been given to the molecular basis of the interaction between FGF-1 and the HSGAG. It is known that within the complex, the carbohydrate maintains the same helical structure of free heparin that leads to sulfate groups directed towards opposite directions along the molecular axis. The precise role of single individual interactions remains unclear, as sliding and/or rotating of the saccharide along the binding pocket are possibilities difficult to discard. The HSGAG binding pocket can be subdivided into two regions, the main one can accommodate a trisaccharide, while the other binds a disaccharide. We have studied and analyzed the interaction between FGF-1 and a library of trisaccharides by STD-NMR and selective longitudinal relaxation rates. The library of trisaccharides corresponds to the heparin backbone and it has been designed to interact with the main subsite of the protein.

Conditional Deletion of Bmal1 Accentuates Microvascular and Macrovascular Injury.

The brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein (BMAL)-1 constitutes a major transcriptional regulator of the circadian clock. Here, we explored the impact of conditional deletion of Bmal1 in endothelium and hematopoietic cells in murine models of microvascular and macrovascular injury. We used two models of Bmal1fx/fx;Tek-Cre mice, a retinal ischemia/reperfusion model and a neointimal hyperplasia model of the femoral artery. Eyes were enumerated for acellular capillaries and were stained for oxidative damage markers using nitrotyrosine immunohistochemistry. LSK (lineage-negative, stem cell antigen-1-positive, c-Kit-positive) cells were quantified and proliferation assessed. Hematopoiesis is influenced by innervation to the bone marrow, which we assessed using IHC analysis. The number of acellular capillaries increased threefold, and nitrotyrosine staining increased 1.5-fold, in the retinas of Bmal1fx/fx;Tek-Cre mice. The number of LSK cells from the Bmal1fx/fx;Tek-Cre mice decreased by 1.5-fold and was accompanied by a profound decrease in proliferative potential. Bmal1fx/fx;Tek-Cre mice also exhibited evidence of bone marrow denervation, demonstrating a loss of neurofilament-200 staining. Injured femoral arteries showed a 20% increase in neointimal hyperplasia compared with similarly injured wild-type controls. Our study highlights the importance of the circadian clock in maintaining vascular homeostasis and demonstrates that specific deletion of BMAL1 in endothelial and hematopoietic cells results in phenotypic features similar to those of diabetes.

Fragment-Based NMR Study of the Conformational Dynamics in the bHLH Transcription Factor Ascl1.

The Achaete-scute homolog 1 (Ascl1) protein regulates a large subset of genes that leads neuronal progenitor cells to distinctive differentiation pathways during human brain development. Although it is well known that Ascl1 binds DNA as a homo- or heterodimer via its basic helix-loop-helix (bHLH) motif, little is known about the conformational sampling properties of the DNA-free full-length protein, and in particular about the bHLH domain-flanking N- and C-terminal segments, which are predicted to be highly disordered in solution. The structural heterogeneity, low solubility, and high aggregation propensity of Ascl1 in aqueous buffer solutions make high-resolution studies of this protein a challenging task. Here, we have adopted a fragment-based strategy that allowed us to obtain high-quality NMR data providing, to our knowledge, the first comprehensive high-resolution information on the structural propensities and conformational dynamics of Ascl1. The emerging picture is that of an overall extended and highly dynamic polypeptide chain comprising three helical segments and lacking persistent long-range interactions. We also show that the C-terminal helix of the bHLH domain is involved in intermolecular interactions, even in the absence of DNA. Our results contribute to a better understanding of the mechanisms of action that govern the regulation of proneural transcription factors.