Epigenetic effects of folate and related B vitamins on brain health throughout life: Scientific substantiation and translation of the evidence for health improvement strategies.

Suboptimal status of folate and/or interrelated B vitamins (B12 , B6 and riboflavin) can perturb one-carbon metabolism and adversely affect brain development in early life and brain function in later life. Human studies show that maternal folate status during pregnancy is associated with cognitive development in the child, whilst optimal B vitamin status may help to prevent cognitive dysfunction in later life. The biological mechanisms explaining these relationships are not clear but may involve folate-related DNA methylation of epigenetically controlled genes related to brain development and function. A better understanding of the mechanisms linking these B vitamins and the epigenome with brain health at critical stages of the lifecycle is necessary to support evidence-based health improvement strategies. The EpiBrain project, a transnational collaboration involving partners in the United Kingdom, Canada and Spain, is investigating the nutrition-epigenome-brain relationship, particularly focussing on folate-related epigenetic effects in relation to brain health outcomes. We are conducting new epigenetics analysis on bio-banked samples from existing well-characterised cohorts and randomised trials conducted in pregnancy and later life. Dietary, nutrient biomarker and epigenetic data will be linked with brain outcomes in children and older adults. In addition, we will investigate the nutrition-epigenome-brain relationship in B vitamin intervention trial participants using magnetoencephalography, a state-of-the-art neuroimaging modality to assess neuronal functioning. The project outcomes will provide an improved understanding of the role of folate and related B vitamins in brain health, and the epigenetic mechanisms involved. The results are expected to provide scientific substantiation to support nutritional strategies for better brain health across the lifecycle.

Three-item loneliness scale: psychometric properties and normative data of the Spanish version.

Loneliness is a common social phenomenon across countries with negative effects in health. Thus, the measurement of loneliness is of paramount importance. The Three-Item Loneliness Scale (TILS) was designed to be used in large-population surveys as a quick measure of loneliness. The aim of this study is to provide a Spanish validation of the TILS. A representative sample of the Spanish population (N = 1951) was used. We analysed the psychometric properties, factor structure, and distribution demographics characteristics of the Spanish TILS. Analyses showed differences regarding age, gender, educational level, employment status, household composition and annual gross income in line with previous literature. The confirmatory factor analysis revealed a unifactorial structure, with significant moderate correlations between the TILS and depression, anxiety, paranoia and well-being, and mild significant correlations with belongingness and mistrust. The internal consistency of the Spanish TILS was good (α = 0.82). Our study indicates that the TILS is a valid and reliable measure of loneliness in the Spanish population. Loneliness is a modern epidemic and a precursor of mental and health problems that reduced the quality of life. Therefore, it is important to have reliable measures of this phenomenon.

Empeoramiento clínico paradójico en una crisis asmática moderada-grave / Paradoxical clinical worsening in a moderate-severe asthmatic attack

La reagudización o crisis asmática es uno de los motivos de consulta más frecuentes en las consultas de Atención Primaria y en los servicios de urgencias pediátricas. Se trata de una patología con un algoritmo de actuación y de tratamiento según la gravedad bien establecido, con fármacos con un buen perfil de seguridad para la edad pediátrica. Se presenta un caso con mala respuesta inicial que ilustra un efecto paradójico del salbutamol (AU)

Asthma exacerbations are among the most frequent reasons for paediatric primary care and emergency care visits. Asthma is a disease with a well-established management and treatment algorithm based on severity, and drugs with a good safety profile for the paediatric population are available for its treatment. We present a case with a poor inital response illustrating a paradoxical reaction to salbutamol. (AU)

Larvicidal activity of the skin secretion of Rhinella marina and Rhaebo guttatus (Anura: Bufonidae) against the Brazilian malaria vector, Anopheles darlingi (Diptera: Culicidae)

@#Malaria, a mosquito-borne disease, is caused by protozoa of the genus Plasmodium and constitutes a serious public health problem. Because current insecticides used to control malaria face resistance due to continuous use, new alternatives are prompted. Considering this context, and the insecticidal potential of vertebrate venoms/secretions, crude and methanolic extracts from two frog species were tested as larvicides against Anopheles darlingi. Skin secretions of Rhinella marina and Rhaebo guttatus were obtained by manual stimulation. Then, methanol was added to obtain steroidal fractions from both venoms. Mosquitos were captured in suburban areas of Porto Velho and An. darlingi females were later fed with blood and stimulated to oviposit. The larvae were fed with fish food until the 3rd and 4th instars. For the larvicidal assays, crude secretions and methanolic fractions of both frog species were evaluated, and larvae mortality was recorded after 48 hours. Crude extracts and steroidal fractions from both species had larvicidal effects, with an LC50 of 127.5 and 133 ppm for the crude extract and steroidal fraction of R. marina, and an LC50 of 37.5 and 35.8 ppm for the crude extract and steroidal secretion of R. guttatus, respectively. The present work reports for the first time the larvicidal effects of the skin secretions from bufonid species occurring in the western Amazon region. Further studies should be carried out to investigate the purified components responsible for the observed activity.

Antimyotoxic Activity of Synthetic Peptides Derived from Bothrops atrox Snake Gamma Phospholipase A2 Inhibitor Selected by Virtual Screening.

BACKGROUND: Several studies have aimed to identify molecules that inhibit the toxic actions of snake venom phospholipases A2 (PLA2s). Studies carried out with PLA2 inhibitors (PLIs) have been shown to be efficient in this assignment. OBJECTIVE: This work aimed to analyze the interaction of peptides derived from Bothrops atrox PLIγ (atPLIγ) with a PLA2 and to evaluate the ability of these peptides to reduce phospholipase and myotoxic activities. METHODS: Peptides were subjected to molecular docking with a homologous Lys49 PLA2 from B. atrox venom modeled by homology. Phospholipase activity neutralization assay was performed with BthTX-II and different ratios of the peptides. A catalytically active and an inactive PLA2 were purified from the B. atrox venom and used together in the in vitro myotoxic activity neutralization experiments with the peptides. RESULTS: The peptides interacted with amino acids near the PLA2 hydrophobic channel and the loop that would be bound to calcium in Asp49 PLA2. They were able to reduce phospholipase activity and peptides DFCHNV and ATHEE reached the highest reduction levels, being these two peptides the best that also interacted in the in silico experiments. The peptides reduced the myotubes cell damage with a highlight for the DFCHNV peptide, which reduced by about 65%. It has been suggested that myotoxic activity reduction is related to the sites occupied in the PLA2 structure, which could corroborate the results observed in molecular docking. CONCLUSION: This study should contribute to the investigation of the potential of PLIs to inhibit the toxic effects of PLA2s.

Pharmacokinetic variations of nimesulide in mongrel dogs according to age.

The purpose of this research was to investigate the possible variations to the pharmacokinetics of nimesulide by the effect of age using an animal model. An experimental, analytical, prospective and longitudinal study in five dogs, from birth to 730 days of age was carried out. Nimesulide blood levels were measured in different months; concentrations were determined by HPLC-UV. Pharmacokinetic parameters were calculated by using the WinNonlin software. There were statistically significant differences (p <0.05) in most of the pharmacokinetic parameters between study of 6 months against the other three studies in different ages. Changes in the pharmacokinetic parameters of nimesulide as a result of age, are determined by the growing and maturation of the animals. Resulting data suggest that nimesulide can be used safely as a long-term analgesic in dogs, but, the dosing regimens in humans should be different when administered at early age.

Guía de práctica clínica para la evaluación y el manejo de la hemorragia digestiva alta en el Seguro Social del Perú (EsSalud) / Clinical practice guideline for the evaluation and management of upper gastrointestinal bleeding for the Peruvian Social Security (EsSalud)

Objetivo: Proveer recomendaciones clínicas basadas en evidencia para la evaluación y el manejo de pacientes con hemorragia digestiva alta (HDA) en el seguro social del Perú (EsSalud). Materiales y métodos: Se conformó un grupo elaborador local (GEG-Local) conformado por especialistas en gastroenterología y metodólogos. El GEG-Local formuló 11 preguntas clínicas a ser respondidas por la presente guía de práctica clínica (GPC). Se buscaron y seleccionaron GPC de HDA publicadas a partir del 2012, que respondieran a las preguntas planteadas y obtuvieran un puntaje mayor a 60% en los dominios 1 y 3 del instrumento Appraisal of Guidelines for Research and Evaluation II (AGREE-II). Durante septiembre del 2017 se realizaron búsquedas bibliográficas en Pubmed, para actualizar 9 preguntas clínicas de las GPC preseleccionadas, y para responder 2 preguntas de novo. La calidad de la evidencia fue evaluada usando la metodología Grading of Recommendations Assessment, Development, and Evaluation (GRADE). En reuniones de trabajo periódicas, el GEG-Local revisó la evidencia y formuló las recomendaciones, los puntos de buenas prácticas clínicas y el flujograma de evaluación y manejo, usando la metodología GRADE Finalmente, la GPC fue aprobada con Resolución N° 80-IETSI-ESSALUD-2017. Resultados: La presente GPC abordó 11 preguntas clínicas, divididas en cuatro temas: valoración de riesgo, manejo inicial, manejo de HDA no variceal, y manejo de HDA variceal. En base a dichas preguntas se formularon 10 recomendaciones (7 recomendaciones fuertes y 3 recomendaciones débiles), 24 puntos de buena práctica clínica, y 2 flujogramas. Conclusión: Este artículo es el resumen de la GPC de EsSalud, en la cual se valoró la evidencia científica disponible sobre evaluación y manejo de HDA.

Objective: To provide evidence-based clinical recommendations for the evaluation and management of patients with upper gastrointestinal bleeding (UGB) in the Peruvian Social Security (EsSalud). Materials and methods: A local guideline development group (local GDG) was established, including specialists in gastroenterology and methodologists. The local GDG formulated 11 clinical questions to be answered by this clinical practice guide (CPG). We searched and selected CPG of UGB published from 2012, which answered the posed questions and obtained a score higher than 60% in domains 1 and 3 of the Appraisal of Guidelines for Research and Evaluation II (AGREE-II) tool. During September 2017, bibliographic searches were conducted in Pubmed, to update 9 clinical questions of the preselected CPGs, and to answer 2 de novo questions. The quality of the evidence was evaluated using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) methodology. In periodic work meetings, the local GDG reviewed the evidence and formulated the recommendations, points of good clinical practice and the flowchart of evaluation and management, using the GRADE methodology Lastly, the CPG was approved with Resolución N° 80-IETSI-ESSALUD-2017. Results: This CPG addressed 11 clinical questions, divided into four themes: risk assessment, initial management, management of non-variceal UGB, and management of variceal UGB. Based on these questions, 10 recommendations (7 strong recommendations and 3 weak recommendations), 24 points of good clinical practice, and 2 flow charts were formulated. Conclusion: This article is the summary of the EsSalud' CPG, where the available scientific evidence on evaluation and management of UGB was evaluated.

Exploring and understanding the functional role, and biochemical and structural characteristics of an acidic phospholipase A2, AplTx-I, purified from Agkistrodon piscivorus leucostoma snake venom.

Phospholipases A2 (PLA2s) constitute a class of extensively studied toxins, isolated from snake venoms. Basic PLA2 isoforms mediate various toxicological effects, while the acidic isoforms generally have higher enzymatic activities, but do not promote evident toxic effects. The functions of these acidic isoforms in snake venoms are still not completely understood and more studies are needed to characterize the biological functions and diversification of acidic toxins in order to justify their abundant presence in these secretions. Recently, Lomonte and collaborators demonstrated, in a proteomic and toxicological study, high concentrations of PLA2s in the venom of Agkistrodon piscivorus leucostoma. We have, herein, purified and characterized an acidic PLA2 from this snake venom, denominated AplTx-I, in order to better understand its biochemical and structural characteristics, as well as its biological effects. AplTx-I was purified using two chromatographic steps, in association with enzymatic and biological assays. The acidic toxin was found to be one of the most abundant proteins in the venom of A. p. leucostoma; the protein was monomeric with a molecular mass of 13,885.8 Da, as identified by mass spectrometry ESI-TOF and electrophoresis. The toxin has similar primary and tridimensional structures to those of other acidic PLA2s, a theoretical and experimental isoelectric point of ≈5.12, and a calcium-dependent enzyme activity of 25.8985 nM/min/mg, with maximum values at 37 °C and pH 8.0. Despite its high enzymatic activity on synthetic substrate, AplTx-I did not induce high or significant myotoxic, coagulant, anticoagulant, edema, neuromuscular toxicity in mouse phrenic nerve-diaphragm preparations or antibacterial activities. Interestingly, AplTx-I triggered a high and selective neuromuscular toxicity in chick biventer cervicis preparations. These findings are relevant to provide a deeper understanding of the pharmacology, role and diversification of acidic phospholipase A2 isoforms in snake venoms.

Snake Venom Peptides and Low Mass Proteins: Molecular Tools and Therapeutic Agents.

Snake venoms are natural sources of biologically active molecules that are able to act selectively and specifically on different cellular targets, modulating physiological functions. Thus, these mixtures, composed mainly of proteins and peptides, provide ample and challenging opportunities and a diversified molecular architecture to design and develop tools and agents of scientific and therapeutic interest. Among these components, peptides and small proteins play diverse roles in numerous physiological processes, exerting a wide range of pharmacological activities, such as antimicrobial, antihypertensive, analgesic, antitumor, analgesic, among others. The pharmaceutical and cosmetic industries have recognized the huge potential of these privileged frameworks and believe them to be a promising alternative to contemporary drugs. A number of natural or synthetic peptides from snake venoms have already found preclinical or clinical applications for the treatment of pain, hypertension, cardiovascular diseases and aging skin. A well-known example is captopril, whose natural peptide precursor was isolated from Bothrops jararaca snake venom, which is a peptide-based drug that inhibits the angiotensin-converting enzyme, producing an anti-hypertensive effect. The present review looks at the main peptides (natriuretic peptides, bradykinin-potentiating peptides and sarafotoxins) and low mass proteins (crotamine, disintegrins and three-Finger toxins) from snake venoms, as well as synthetic peptides inspired by them, describing their biochemical, structural and physiological features, as well as their applications as research tools and therapeutic agents.

Administración accidental de aceite puro esencial de arbol de té (Melaleuca alternifolia) en lactantes / Accidental pure essential tea tree oil (Melaleuca alternifolia) administration in two infants

El aceite de árbol de té es una sustancia que se obtiene mediante la destilación de las hojas y ramas frescas del árbol Melaleuca alternifolia. En los últimos años se ha popularizado su uso sobre todo como agente antiinfeccioso tópico en una gran variedad de enfermedades. Se trata de una sustancia natural que tiene un potencial efecto tóxico demostrado sobre todo si se ingiere. Presentamos dos casos de pacientes atendidos en nuestro Servicio de Urgencias tras administración e ingestión accidental de aceite esencial de árbol de té, en ambos casos los padres confundieron el envase con el de la vitamina D. La atención de estos pacientes nos llevó a realizar una búsqueda bibliográfica de casos publicados de intoxicación por ingesta de aceite de árbol de té y a poner de manifiesto la ausencia de advertencias de seguridad en los envases de aceite de árbol de té que actualmente se comercializan en nuestro país (AU)

Tea tree oil is an essential oil obtained by steam distillation of the leaves and terminal branches of Melaleuca alternifolia. In recent years, it has become popular as an antimicrobial agent against a large number of diseases. It is a natural substance that has a potential toxic effect especially if ingested. We report two cases of infants who came to our Emergency Department because they were accidentally given tea tree oil. In both cases parents mistook the tea tree oil bottle with the D vitamin bottle. The care of these patients led us to perform a literature search of published cases of tea tree oil poisoning and highlight the absence of safety warnings on tea tree oil packages currently marketed in our country (AU)

CoaTx-II, a new dimeric Lys49 phospholipase A2 from Crotalus oreganus abyssus snake venom with bactericidal potential: Insights into its structure and biological roles.

Snake venoms are rich and intriguing sources of biologically-active molecules that act on target cells, modulating a diversity of physiological functions and presenting promising pharmacological applications. Lys49 phospholipase A2 is one of the multifunctional proteins present in these complex secretions and, although catalytically inactive, has a variety of biological activities, including cytotoxic, antibacterial, inflammatory, antifungal activities. Herein, a Lys49 phospholipase A2, denominated CoaTx-II from Crotalus oreganus abyssus, was purified and structurally and pharmacologically characterized. CoaTx-II was isolated with a high degree of purity by a combination of two chromatographic steps; molecular exclusion and reversed-phase high performance liquid chromatography. This toxin is dimeric with a mass of 13868.2 Da (monomeric form), as determined by mass spectrometry. CoaTx-II is rich in Arg and Lys residues and displays high identity with other Lys49 PLA2 homologues, which have high isoelectric points. The structural model of dimeric CoaTx-II shows that the toxin is non-covalently stabilized. Despite its enzymatic inactivity, in vivo CoaTx-II caused local muscular damage, characterized by increased plasma creatine kinase and confirmed by histological alterations, in addition to an inflammatory activity, as demonstrated by mice paw edema induction and pro-inflammatory cytokine IL-6 elevation. CoaTx-II also presents antibacterial activity against gram negative (Pseudomonas aeruginosa 31NM, Escherichia coli ATCC 25922) and positive (Staphyloccocus aureus BEC9393 and Rib1) bacteria. Therefore, data show that this newly purified toxin plays a central role in mediating the degenerative events associated with envenomation, in addition to demonstrating antibacterial properties, with potential for use in the development of strategies for antivenom therapy and combating antibiotic-resistant bacteria.

Biochemical and functional studies of ColTx-I, a new myotoxic phospholipase A2 isolated from Crotalus oreganus lutosus (Great Basin rattlesnake) snake venom.

Commonly, phospholipases A2 (PLA2s) play key roles in the pathogenesis of the local tissue damage characteristic of crotaline and viperine snake envenomations. Crotalus oreganus lutosus snake venom has not been extensively studied; therefore, the characterization of its components represents a valuable biotechnological tool for studying pathophysiological processes of envenoming and for gaining a deeper understanding of its biological effects. In this study, for the first time, a basic PLA2 myotoxin, ColTx-I, was purified from C. o. lutosus through two chromatographic steps. ColTx-I is monomeric with calculated molecular mass weight (Mw) of 14,145 Da and a primary structure closely related to basic PLA2s from viperid venoms. The pure enzyme has a specific activity of 15.87 ± 0.65 nmol/min/mg at optimal conditions (pH 8.0 and 37 °C). ColTx-I activity was found to be dependent on Ca(2+), as its substitution by other ionic species as well as the addition of chelating agents significantly reduced its phospholipase activity. In vivo, ColTx-I triggered dose-dependent inflammatory responses, measured using the paw edema model, with an increase in IL-6 levels, systemic and local myotoxicity, characterized by elevated plasma creatine kinase activity. ColTx-I induced a complex series of degenerative events associated with edema, inflammatory infiltrate and skeletal muscle necrosis. These biochemical and functional results suggest that ColTx-I, a myotoxic and inflammatory mediator, plays a relevant role in C. o. lutosus envenomation. Thus, detailed studies on its mechanism of action, such as evaluating the synergism between ColTx-I and other venom components may reveal targets for the development of more specific and effective therapies.

Análisis de atención inicial y manejo del traumatismo craneoencefálico estable en Urgencias de Pediatría. Estudio comparativo / Analysis of initial care and management of stable cranioencephalic traumatism in Pediatric Emergency Department. A comparative study

El traumatismo craneoencefálico (TCE) constituye un motivo de consulta frecuente en Urgencias de Pediatría; la mayoría de tipo leve. Tras su valoración con el triángulo de evaluación pediátrica y clasificación del nivel de urgencia, se debe aplicar un protocolo de actuación que establezca su adecuado manejo y solicitud de prueba de imagen. El objetivo del estudio es analizar las características de dos grupos de pacientes valorados con 4 años de diferencia, establecer su valoración inicial, pruebas de imagen y necesidad de observación hospitalaria. Resultados. Total 169 episodios (prevalencia 2,6%). Año 2010: 92 episodios, 90 pacientes. Año 2014: 77 episodios, 76 pacientes. En ambos años predomina el sexo masculino, el motivo de consulta más prevalente es el TCE aislado, y cuando asocia clínica predominan los vómitos, y en la exploración la existencia de herida externa. El registro de constantes de frecuencia cardiaca y presión arterial se ha incrementado significativamente. Se ha reducido el porcentaje de radiografías de cráneo simple realizadas de forma global en 1,4% y en la franja de edad entre 1 y 2 años en un 7,1%. El porcentaje de tomografía axial computarizada craneal (TAC) fue similar en ambos años. Precisó observación un 13% en 2010 y 9,1% en 2014. Discusión. Es imprescindible conseguir un equilibrio en la valoración urgente del TCE estable que permita reducir la radiación aplicada al paciente en forma de radiología convencional, mantener la adecuación de la indicación de la TAC craneal ajustada y su estancia en forma de observación hospitalaria

Cranioencephalic traumatism (CET) is a cause of frequent medical visits in the Pediatric Emergency Department, most of them being mild. After evaluation with the pediatric assessment triangle and classification on an emergency level, an action protocol should be applied that establishes its adequate management and request for imaging test. The purpose of the study is to analyze the characteristics of two groups of patients evaluated with a different of 4 years, to establish their initial evaluation, imaging tests and need for hospital observation. Results. Total 169 episodes (prevalence 2.6%). Year 2010: 92 episodes, 90 patients. Year 2014: 77 episodes, 76 patients. In both years, there was a predominance of male gender, the most prevalent reason for the visit was isolated CET, and when symptoms were associated, vomiting clinically predominated, while in the physical exam, the external wound predominated. The recording of the vital signs of heart rate and blood pressure has significantly increased. The percentage of simple brain x-rays performed globally has reduced by 1.4% and the age range between 1 and 2 years by 7.1%. The percentage of cranial computed tomography was similar in both years. A total of 13% required observation in 2010 and 9.1% in 2014. Discussion. It is essential to achieve a balance in urgent assessment of stable CET that would make it possible to reduce the radiation applied to the patient in form of conventional radiology, to maintain adaptation of the indication of the cranial CT scan indication and the patients stay in form of hospital observation

Alkylation of histidine residues of Bothrops jararacussu venom proteins and isolated phospholipases A2: a biotechnological tool to improve the production of antibodies.

Crude venom of Bothrops jararacussu and isolated phospholipases A2 (PLA2) of this toxin (BthTX-I and BthTX-II) were chemically modified (alkylation) by p-bromophenacyl bromide (BPB) in order to study antibody production capacity in function of the structure-function relationship of these substances (crude venom and PLA2 native and alkylated). BthTX-II showed enzymatic activity, while BthTX-I did not. Alkylation reduced BthTX-II activity by 50% while this process abolished the catalytic and myotoxic activities of BthTX-I, while reducing its edema-inducing activity by about 50%. Antibody production against the native and alkylated forms of BthTX-I and -II and the cross-reactivity of antibodies to native and alkylated toxins did not show any apparent differences and these observations were reinforced by surface plasmon resonance (SPR) data. Histopathological analysis of mouse gastrocnemius muscle sections after injection of PBS, BthTX-I, BthTX-II, or both myotoxins previously incubated with neutralizing antibody showed inhibition of the toxin-induced myotoxicity. These results reveal that the chemical modification of the phospholipases A2 (PLA2) diminished their toxicity but did not alter their antigenicity. This observation indicates that the modified PLA2 may provide a biotechnological tool to attenuate the toxicity of the crude venom, by improving the production of antibodies and decreasing the local toxic effects of this poisonous substance in animals used to produce antivenom.