Impact of JQ1 treatment on seizures, hippocampal gene expression, and gliosis in a mouse model of temporal lobe epilepsy.

Epilepsy is a neurological disease characterised by recurrent seizures with complex aetiology. Temporal lobe epilepsy, the most common form in adults, can be acquired following brain insults including trauma, stroke, infection or sustained status epilepticus. The mechanisms that give rise to the formation and maintenance of hyperexcitable networks following acquired insults remain unknown, yet an extensive body of literature points towards persistent gene and epigenomic dysregulation as a potential mediator of this dysfunction. While much is known about the function of specific classes of epigenetic regulators (writers and erasers) in epilepsy, much less is known about the enzymes, which read the epigenome and modulate gene expression accordingly. Here, we explore the potential role for the epigenetic reader bromodomain and extra-terminal domain (BET) proteins in epilepsy. Using the intra-amygdala kainic acid model of temporal lobe epilepsy, we initially identified widespread dysregulation of important epigenetic regulators including EZH2 and REST as well as altered BRD4 expression in chronically epileptic mice. BRD4 activity was also notably affected by epilepsy-provoking insults as seen by elevated binding to and transcriptional regulation of the immediate early gene Fos. Despite influencing early aspects of epileptogenesis, blocking BET protein activity with JQ1 had no overt effects on epilepsy development in mice but did alter glial reactivity and influence gene expression patterns, promoting various neurotransmitter signalling mechanisms and inflammatory pathways in the hippocampus. Together, these results confirm that epigenetic reader activity is affected by epilepsy-provoking brain insults and that BET activity may exert cell-specific actions on inflammation in epilepsy.

The Maillard reactions: Pathways, consequences, and control.

The century old Maillard reactions continue to draw the interest of researchers in the fields of Food Science and Technology, and Health and Medical Sciences. This chapter seeks to simplify and update this highly complicated, multifaceted topic. The simple nucleophilic attack of an amine onto a carbonyl group gives rise to a series of parallel and subsequent reactions, occurring simultaneously, resulting into a vast array of low and high mass compounds. Recent research has focused on: (1) the formation and transformation of α-dicarbonyl compounds, highly reactive intermediates which are essential in the development of the desired color and flavor of foods, but also lead to the production of the detrimental advanced glycation end products (AGEs); (2) elucidation of the structures of melanoidins in different foods and their beneficial effects on human health; and (3) harmful effects of AGEs on human health. Considering that MRs have both positive and negative consequences, their control to accentuate the former and to mitigate the latter, is also being conscientiously investigated with the use of modern techniques and technology.

Differential microRNA editing may drive target pathway switching in human temporal lobe epilepsy.

MicroRNAs have emerged as important regulators of the gene expression landscape in temporal lobe epilepsy. The mechanisms that control microRNA levels and influence target choice remain, however, poorly understood. RNA editing is a post-transcriptional mechanism mediated by the adenosine acting on RNA (ADAR) family of proteins that introduces base modification that diversifies the gene expression landscape. RNA editing has been studied for the mRNA landscape but the extent to which microRNA editing occurs in human temporal lobe epilepsy is unknown. Here, we used small RNA-sequencing data to characterize the identity and extent of microRNA editing in human temporal lobe epilepsy brain samples. This detected low-to-high editing in over 40 of the identified microRNAs. Among microRNA exhibiting the highest editing was miR-376a-3p, which was edited in the seed region and this was predicted to significantly change the target pool. The edited form was expressed at lower levels in human temporal lobe epilepsy samples. We modelled the shift in editing levels of miR-376a-3p in human-induced pluripotent stem cell-derived neurons. Reducing levels of the edited form of miR-376a-3p using antisense oligonucleotides resulted in extensive gene expression changes, including upregulation of mitochondrial and metabolism-associated pathways. Together, these results show that differential editing of microRNAs may re-direct targeting and result in altered functions relevant to the pathophysiology of temporal lobe epilepsy and perhaps other disorders of neuronal hyperexcitability.

Comprehensive Update on Carotenoid Colorants from Plants and Microalgae: Challenges and Advances from Research Laboratories to Industry.

The substitution of synthetic food dyes with natural colorants continues to be assiduously pursued. The current list of natural carotenoid colorants consists of plant-derived annatto (bixin and norbixin), paprika (capsanthin and capsorubin), saffron (crocin), tomato and gac fruit lycopene, marigold lutein, and red palm oil (α- and ß-carotene), along with microalgal Dunaliella ß-carotene and Haematococcus astaxanthin and fungal Blakeslea trispora ß-carotene and lycopene. Potential microalgal sources are being sought, especially in relation to lutein, for which commercial plant sources are lacking. Research efforts, manifested in numerous reviews and research papers published in the last decade, have been directed to green extraction, microencapsulation/nanoencapsulation, and valorization of processing by-products. Extraction is shifting from conventional extraction with organic solvents to supercritical CO2 extraction and different types of assisted extraction. Initially intended for the stabilization of the highly degradable carotenoids, additional benefits of encapsulation have been demonstrated, especially the improvement of carotenoid solubility and bioavailability. Instead of searching for new higher plant sources, enormous effort has been directed to the utilization of by-products of the fruit and vegetable processing industry, with the application of biorefinery and circular economy concepts. Amidst enormous research activities, however, the gap between research and industrial implementation remains wide.

Brain cell-specific origin of circulating microRNA biomarkers in experimental temporal lobe epilepsy.

The diagnosis of epilepsy is complex and challenging and would benefit from the availability of molecular biomarkers, ideally measurable in a biofluid such as blood. Experimental and human epilepsy are associated with altered brain and blood levels of various microRNAs (miRNAs). Evidence is lacking, however, as to whether any of the circulating pool of miRNAs originates from the brain. To explore the link between circulating miRNAs and the pathophysiology of epilepsy, we first sequenced argonaute 2 (Ago2)-bound miRNAs in plasma samples collected from mice subject to status epilepticus induced by intraamygdala microinjection of kainic acid. This identified time-dependent changes in plasma levels of miRNAs with known neuronal and microglial-cell origins. To explore whether the circulating miRNAs had originated from the brain, we generated mice expressing FLAG-Ago2 in neurons or microglia using tamoxifen-inducible Thy1 or Cx3cr1 promoters, respectively. FLAG immunoprecipitates from the plasma of these mice after seizures contained miRNAs, including let-7i-5p and miR-19b-3p. Taken together, these studies confirm that a portion of the circulating pool of miRNAs in experimental epilepsy originates from the brain, increasing support for miRNAs as mechanistic biomarkers of epilepsy.

Comprehensive review on carotenoid composition: Transformations during processing and storage of foods.

Stimulated by their multifaceted functions and actions, carotenoids have been among the most investigated food components, producing a voluminous, complicated, and sometimes inconsistent literature. This review puts into context developments in the last decade to have a comprehensive current knowledge on these valuable food constituents. Carotenoid analysis continues to show the wide biodiversity of carotenogenic foods and the many factors that affect the composition. Because of their instability, subject to multiple influencing factors, retention of carotenoids during processing and storage of food has been a daunting task. Since thermal processing may result in substantial carotenoid losses, thermal processes that are much faster than the conventional methods and nonthermal processing have been introduced. The processing conditions of nonthermal processing should, however, be well established so that microbial and enzymatic inactivation is achieved while maintaining nutrients and bioactive compounds. Instead of losses, higher carotenoid levels and bioaccessibility are sometimes reported for both thermal and nonthermal processing, attributed to greater extractability of carotenoids during analysis and greater release from the food matrix during digestion. Carotenoids differ markedly in their susceptibility to degradation, the epoxycarotenoids being most degradable. Results are mixed, however, in relation to the comparative stability of hydroxycarotenoids and carotenes. E-Z isomerization at sterically unhindered double bonds is now well documented. There is also more information about oxidative degradation, although more work is needed on this topic. It consists of epoxidation, cleavage to apocarotenoids and finally fragmention to low mass compounds. Enzymatic and non-enzymatic cleavage of carotenoids forms important aroma compounds but can also produce off-flavor.

Anti-seizure effects of JNJ-54175446 in the intra-amygdala kainic acid model of drug-resistant temporal lobe epilepsy in mice.

There remains a need for new drug targets for treatment-resistant temporal lobe epilepsy. The ATP-gated P2X7 receptor coordinates neuroinflammatory responses to tissue injury. Previous studies in mice reported that the P2X7 receptor antagonist JNJ-47965567 suppressed spontaneous seizures in the intraamygdala kainic acid model of epilepsy and reduced attendant gliosis in the hippocampus. The drug-resistance profile of this model is not fully characterised, however, and newer P2X7 receptor antagonists with superior pharmacokinetic profiles have recently entered clinical trials. Using telemetry-based continuous EEG recordings in mice, we demonstrate that spontaneous recurrent seizures in the intraamygdala kainic acid model are refractory to the common anti-seizure medicine levetiracetam. In contrast, once-daily dosing of JNJ-54175446 (30 mg/kg, intraperitoneal) resulted in a significant reduction in spontaneous recurrent seizures which lasted several days after the end of drug administration. Using a combination of immunohistochemistry and ex vivo radiotracer assay, we find that JNJ-54175446-treated mice at the end of recordings display a reduction in astrogliosis and altered microglia process morphology within the ipsilateral CA3 subfield of the hippocampus, but no difference in P2X7 receptor surface expression. The present study extends the characterisation of the drug-resistance profile of the intraamygdala kainic acid model in mice and provides further evidence that targeting the P2X7 receptor may have therapeutic applications in the treatment of temporal lobe epilepsy.

MicroRNA inhibition using antimiRs in acute human brain tissue sections.

Antisense inhibition of microRNAs is an emerging preclinical approach to pharmacoresistant epilepsy. A leading candidate is an "antimiR" targeting microRNA-134 (ant-134), but testing to date has used rodent models. Here, we develop an antimiR testing platform in human brain tissue sections. Brain specimens were obtained from patients undergoing resective surgery to treat pharmacoresistant epilepsy. Neocortical specimens were submerged in modified artificial cerebrospinal fluid (ACSF) and dissected for clinical neuropathological examination, and unused material was transferred for sectioning. Individual sections were incubated in oxygenated ACSF, containing either ant-134 or a nontargeting control antimiR, for 24 h at room temperature. RNA integrity was assessed using BioAnalyzer processing, and individual miRNA levels were measured using quantitative reverse transcriptase polymerase chain reaction. Specimens transported in ACSF could be used for neuropathological diagnosis and had good RNA integrity. Ant-134 mediated a dose-dependent knockdown of miR-134, with approximately 75% reduction of miR-134 at 1 µmol L-1 and 90% reduction at 3 µmol L-1 . These doses did not have off-target effects on expression of a selection of three other miRNAs. This is the first demonstration of ant-134 effects in live human brain tissues. The findings lend further support to the preclinical development of a therapy that targets miR-134 and offer a flexible platform for the preclinical testing of antimiRs, and other antisense oligonucleotide therapeutics, in human brain.

El aprendizaje y las nuevas tecnologías de la información y las comunicaciones / Learning and the new information and communication technologies

Las nuevas tecnologías de la información y las comunicaciones insertan cambios progresivos en el sistema de enseñanza actual, porque la influencia de la innovación tecnológica propicia nuevas formas de concebir el aprendizaje. Por ello, las concepciones, los procesos y los paradigmas de la actividad docente-educativa han migrado del modelo tradicional al tecnopedagógico de manera paulatina. De este modo, se planteó como objetivo exponer los aspectos relacionados con el aprendizaje, las distintas teorías que lo sustentan y los elementos más significativos que giran en torno a las redes de aprendizaje y aprendizaje en red. El estudio empleó la contrastación de métodos teóricos como el histórico-lógico, y la sistematización teórica para la recolección de información, la construcción, el desarrollo y la conformación final del producto. Los resultados se enfocaron en actualizar los conocimientos teóricos del aprendizaje y las nuevas tecnologías de la información y las comunicaciones, para incentivar a la comunidad de investigadores a que profundizaran sobre el tema. Las nuevas tecnologías de la información y las comunicaciones le imprimen al aprendizaje un carácter más autónomo, e incrementan, a su vez, su carácter social, debido al constante desarrollo de la web, las redes sociales, los ambientes virtuales de aprendizaje y las comunidades en internet. A su vez, el conectivismo ha permitido establecer nuevas formas de concebir la educación, al articular de manera adecuada lo tecnológico con lo pedagógico, mediante un proceso de formación continua del docente en temas de tecnología aplicada a la educación(AU)

The new information and communication technologies add progressive changes in the current teaching system, as long as the influence of technological innovation fosters new ways of conceiving learning. For this reason, conceptions, processes and paradigms of the teaching-educational activity have gradually migrated from the traditional to the techno-pedagogical model. Based on this, the objective was to expose the aspects related to learning, the different theories that support it and the most significant elements related to learning and learning networks. The study used the contrast between theoretical methods, such as the historical-logical, as well as theoretical systematization for gathering information, construction, the development and the final structuring of the product. The results were focused on updating the theoretical knowledge of learning and the new information and communication technologies, in order to encourage the research community to deepen on the subject. The new information and communication technologies give learning a more autonomous character, and increase, in turn, its social character, due to the continuous development in the Web, in social networks and in virtual learning environments and communities in the Internet. In turn, connectivism has made it possible to establish new ways of conceiving education, by adequately articulating the technological with the pedagogical, through a process of continuous training for the professor regarding issues of technology applied to education(AU)

Systemic delivery of antagomirs during blood-brain barrier disruption is disease-modifying in experimental epilepsy.

Oligonucleotide therapies offer precision treatments for a variety of neurological diseases, including epilepsy, but their deployment is hampered by the blood-brain barrier (BBB). Previous studies showed that intracerebroventricular injection of an antisense oligonucleotide (antagomir) targeting microRNA-134 (Ant-134) reduced evoked and spontaneous seizures in animal models of epilepsy. In this study, we used assays of serum protein and tracer extravasation to determine that BBB disruption occurring after status epilepticus in mice was sufficient to permit passage of systemically injected Ant-134 into the brain parenchyma. Intraperitoneal and intravenous injection of Ant-134 reached the hippocampus and blocked seizure-induced upregulation of miR-134. A single intraperitoneal injection of Ant-134 at 2 h after status epilepticus in mice resulted in potent suppression of spontaneous recurrent seizures, reaching a 99.5% reduction during recordings at 3 months. The duration of spontaneous seizures, when they occurred, was also reduced in Ant-134-treated mice. In vivo knockdown of LIM kinase-1 (Limk-1) increased seizure frequency in Ant-134-treated mice, implicating de-repression of Limk-1 in the antagomir mechanism. These studies indicate that systemic delivery of Ant-134 reaches the brain and produces long-lasting seizure-suppressive effects after systemic injection in mice when timed with BBB disruption and may be a clinically viable approach for this and other disease-modifying microRNA therapies.

Factores relacionados con letalidad en pacientes con bacteriemia hospitalizados por patología médica en una institución de tercer nivel en Colombia, 2014-2016 / Case fatality rate-related factors in patients with bacteremia hospitalized due medical conditions in an institution of third level in Colombia, 2014-2016

Resumen Introducción: Las infecciones del torrente sanguíneo son un problema creciente y actualmente son una amenaza para la salud pública. La bacteriemia representa aproximadamente 15% de todas las infecciones nosocomiales y afecta a 1% de los pacientes hospitalizados. Objetivo: Describir las características clínicas, epidemiológicas y microbiológicas de episodios de bacteriemia nosocomial ocurridos en un hospital colombiano. Pacientes y Métodos: Estudio retrospectivo, observacional, de corte transversal, con inclusión de pacientes adultos, hospitalizados por el Servicio de Medicina Interna en el Hospital Universitario de Santander, Bucaramanga, Colombia, durante los años 2014 a 2016. El protocolo fue aprobado por el Comité de Ética en Investigación de la Universidad Industrial de Santander. Resultados: Se revisaron 450 historias clínicas, con 148 pacientes y 182 aislados microbianos. Los antecedentes más frecuentes fueron: hipertensión arterial (46,6%) e infección por VIH (29,7%). El sistema vascular y urinario ocuparon los sitios anatómicos más frecuentes (37,3 y 38,3%, respectivamente). La letalidad fue de 29%. Los patógenos más frecuentemente aislados fueron: Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa (en suma: 49,8%) y Staphylococcus aureus 12,1%. El análisis multivariado mostró relación de la anemia con mortalidad intrahospitalaria (OR = 17,3; IC95% 2,95-102,0). Conclusiones: La bacteriemia es una infección frecuente durante la atención hospitalaria que presenta gran mortalidad. Es destacable el predominio de aislados de enterobacterias multiresistentes. El antecedente de infección por VIH es uno de los más frecuentes el que amerita ser evaluado como grupo de riesgo.

Abstract Background: Bloodstream infections are an increasing problem and currently represent a threat to public health, overcoming diseases such as HIV. Bacteremia accounts for approximately 15% of all nosocomial infections and affects 1% of all hospitalized patients. Aim: To describe the clinical, epidemiological and microbiological characteristic of episodes of nosocomial bacteremia occurring in a Colombian hospital. Methods: Retrospective, observational, cross-sectional study including adult patients, hospitalized in the internal medicine unit at the University Hospital of Santander, Bucaramanga, Colombia, during years 2014 to 2016, who met the criteria of the CDC for bloodstream infection. The protocol was approved by the Hospital Ethics Committee and by the Research Ethics Committee of the Industrial University of Santander. Results: We reviewed 450 clinical records with 148 patients and 182 microbiological isolates. 53% were male. The most frequent comorbidities were: high blood pressure (46.6%), HIV infection (29.7%). The vascular and urinary systems were the most frequent anatomical sites as the source of the infection (respectively 37.3% and 38.3%). Case fatality rate was 29%. The pathogens most frequently isolated were: Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa (globally: 49.8%) and Staphylococcus aureus 12.1%. The multivariate analysis showed a relationship between anemia and in-hospital mortality (OR = 17.3, 95%CI 2.95-102.0). Conclusions: Bacteremia is a frequent infection during hospital care that presents high mortality. It is noteworthy the predominance of Enterobacteriaceae isolates with broad profiles of resistance. The history of HIV infection is one of the most frequent which deserves to be evaluated as a risk group.

Weight change in women with breast cancer (IIIB) after treatment

Introduction: the effect of overweight and obesity in patients with breast cancer has been widely described. Despite the recognition of ethnic differences in these associations, information is still lacking for the Latin American population. Methods: a retrospective, longitudinal cohort study with non-probabilistic sampling. The main objective was to describe how weight behaved after multimodal cancer treatment in women with locally advanced luminal A subtype breast cancer. Results: the average age at cancer diagnosis was 52 years. The average follow-up time was 2.3 years, during which there was a 12.1% rate of recurrence. Most patients were overweight/obese (67.56%), with an average variation of -0.17 kg at the end of follow up. Patients with metastasis had a greater weight loss than those without recurrence (-5.06 kg, p<0.05). Conclusions: overweight and obesity are a prevalent characteristic of locally advanced luminal A breast cancer patients. There was no conclusive evidence of increased risk of metastasis or death related to excess weight in this population. To the contrary, weight loss was a statistically significant characteristic of patients with distal recurrence during follow up, although it was not established as a causal factor.

Introducción: se ha descrito ampliamente el efecto del sobrepeso y la obesidad en pacientes con cáncer de mama. A pesar de que se reconocen diferencias étnicas de dichas asociaciones, aún es insuficiente la información en población latinoamericana. Metodología: estudio longitudinal tipo cohorte retrospectiva, con muestreo no probabilístico. El objetivo principal fue describir el comportamiento del peso de mujeres con cáncer de mama, localmente avanzado del subtipo luminal A después de tratamiento oncológico multimodal. Resultados: la edad promedio al diagnóstico de cáncer fue de 52 años. El tiempo promedio de seguimiento de 2.3 años en los cuales hubo una tasa de recurrencia de 12.1%. La mayoría de las pacientes presentaron sobrepeso/obesidad (67.56%) con una variación promedio de -0.17 Kg al final del seguimiento. Las pacientes con metástasis presentaron una pérdida mayor de peso que su contraparte sin recurrencia (-5.06 Kg, p<0.05). Conclusiones: el sobrepeso y obesidad son una característica muy prevalente en pacientes con cáncer de mama luminal A en estadio localmente avanzado. No hubo evidencia concluyente de aumento de riesgo de metástasis o muerte asociado a exceso de peso en esta población. Al contrario la pérdida de peso fue una característica estadísticamente significativa de pacientes que presentaron recurrencia a distancia durante el seguimiento, sin poderse definir como un factor causal.(Acta Med Colomb 2020; 45. DOI:https://doi.org/10.36104/amc.2020.1563).

Variabilidad del peso en mujeres con cáncer de mama (IIIB) después de tratamiento / Weight change in women after treatment with breast cancer (IIIB)

Resumen Introducción: se ha descrito ampliamente el efecto del sobrepeso y la obesidad en pacientes con cáncer de mama. A pesar de que se reconocen diferencias étnicas de dichas asociaciones, aún es insuficiente la información en población latinoamericana. Metodología: estudio longitudinal tipo cohorte retrospectiva, con muestreo no probabilístico. El objetivo principal fue describir el comportamiento del peso de mujeres con cáncer de mama, localmente avanzado del subtipo luminal A después de tratamiento oncológico multimodal. Resultados: la edad promedio al diagnóstico de cáncer fue de 52 años. El tiempo promedio de seguimiento de 2.3 años en los cuales hubo una tasa de recurrencia de 12.1%. La mayoría de las pacientes presentaron sobrepeso/obesidad (67.56%) con una variación promedio de -0.17 Kg al final del seguimiento. Las pacientes con metástasis presentaron una pérdida mayor de peso que su contraparte sin recurrencia (-5.06 Kg, p<0.05). Conclusiones: el sobrepeso y obesidad son una característica muy prevalente en pacientes con cáncer de mama luminal A en estadio localmente avanzado. No hubo evidencia concluyente de aumento de riesgo de metástasis o muerte asociado a exceso de peso en esta población. Al contrario la pérdida de peso fue una característica estadísticamente significativa de pacientes que presentaron recurrencia a distancia durante el seguimiento, sin poderse definir como un factor causal.(Acta Med Colomb 2020; 45. DOI:https://doi.org/10.36104/amc.2020.1563).

Abstract Introduction: the effect of overweight and obesity in patients with breast cancer has been widely described. Despite the recognition of ethnic differences in these associations, information is still lacking for the Latin American population. Methods: a retrospective, longitudinal cohort study with non-probabilistic sampling. The main objective was to describe how weight behaved after multimodal cancer treatment in women with locally advanced luminal A subtype breast cancer. Results: the average age at cancer diagnosis was 52 years. The average follow-up time was 2.3 years, during which there was a 12.1% rate of recurrence. Most patients were overweight/obese (67.56%), with an average variation of -0.17 kg at the end of follow up. Patients with metastasis had a greater weight loss than those without recurrence (-5.06 kg, p<0.05). Conclusions: overweight and obesity are a prevalent characteristic of locally advanced luminal A breast cancer patients. There was no conclusive evidence of increased risk of metastasis or death related to excess weight in this population. To the contrary, weight loss was a statistically significant characteristic of patients with distal recurrence during follow up, although it was not established as a causal factor.(Acta Med Colomb 2020; 45. DOI:https://doi.org/10.36104/amc.2020.1563).

Genetic deletion of microRNA-22 blunts the inflammatory transcriptional response to status epilepticus and exacerbates epilepsy in mice.

MicroRNAs perform important roles in the post-transcriptional regulation of gene expression. Sequencing as well as functional studies using antisense oligonucleotides indicate important roles for microRNAs during the development of epilepsy through targeting transcripts involved in neuronal structure, gliosis and inflammation. MicroRNA-22 (miR-22) has been reported to protect against the development of epileptogenic brain networks through suppression of neuroinflammatory signalling. Here, we used mice with a genetic deletion of miR-22 to extend these insights. Mice lacking miR-22 displayed normal behaviour and brain structure and developed similar status epilepticus after intraamygdala kainic acid compared to wildtype animals. Continuous EEG monitoring after status epilepticus revealed, however, an accelerated and exacerbated epilepsy phenotype whereby spontaneous seizures began sooner, occurred more frequently and were of longer duration in miR-22-deficient mice. RNA sequencing analysis of the hippocampus during the period of epileptogenesis revealed a specific suppression of inflammatory signalling in the hippocampus of miR-22-deficient mice. Taken together, these findings indicate a role for miR-22 in establishing early inflammatory responses to status epilepticus. Inflammatory signalling may serve anti-epileptogenic functions and cautions the timing of anti-inflammatory interventions for the treatment of status epilepticus.

Genome-wide microRNA profiling of plasma from three different animal models identifies biomarkers of temporal lobe epilepsy.

Epilepsy diagnosis is complex, requires a team of specialists and relies on in-depth patient and family history, MRI-imaging and EEG monitoring. There is therefore an unmet clinical need for a non-invasive, molecular-based, biomarker to either predict the development of epilepsy or diagnose a patient with epilepsy who may not have had a witnessed seizure. Recent studies have demonstrated a role for microRNAs in the pathogenesis of epilepsy. MicroRNAs are short non-coding RNA molecules which negatively regulate gene expression, exerting profound influence on target pathways and cellular processes. The presence of microRNAs in biofluids, ease of detection, resistance to degradation and functional role in epilepsy render them excellent candidate biomarkers. Here we performed the first multi-model, genome-wide profiling of plasma microRNAs during epileptogenesis and in chronic temporal lobe epilepsy animals. From video-EEG monitored rats and mice we serially sampled blood samples and identified a set of dysregulated microRNAs comprising increased miR-93-5p, miR-142-5p, miR-182-5p, miR-199a-3p and decreased miR-574-3p during one or both phases. Validation studies found miR-93-5p, miR-199a-3p and miR-574-3p were also dysregulated in plasma from patients with intractable temporal lobe epilepsy. Treatment of mice with common anti-epileptic drugs did not alter the expression levels of any of the five miRNAs identified, however administration of an anti-epileptogenic microRNA treatment prevented dysregulation of several of these miRNAs. The miRNAs were detected within the Argonuate2-RISC complex from both neurons and microglia indicating these miRNA biomarker candidates can likely be traced back to specific brain cell types. The current studies identify additional circulating microRNA biomarkers of experimental and human epilepsy which may support diagnosis of temporal lobe epilepsy via a quick, cost-effective rapid molecular-based test.

A systems approach delivers a functional microRNA catalog and expanded targets for seizure suppression in temporal lobe epilepsy.

Temporal lobe epilepsy is the most common drug-resistant form of epilepsy in adults. The reorganization of neural networks and the gene expression landscape underlying pathophysiologic network behavior in brain structures such as the hippocampus has been suggested to be controlled, in part, by microRNAs. To systematically assess their significance, we sequenced Argonaute-loaded microRNAs to define functionally engaged microRNAs in the hippocampus of three different animal models in two species and at six time points between the initial precipitating insult through to the establishment of chronic epilepsy. We then selected commonly up-regulated microRNAs for a functional in vivo therapeutic screen using oligonucleotide inhibitors. Argonaute sequencing generated 1.44 billion small RNA reads of which up to 82% were microRNAs, with over 400 unique microRNAs detected per model. Approximately half of the detected microRNAs were dysregulated in each epilepsy model. We prioritized commonly up-regulated microRNAs that were fully conserved in humans and designed custom antisense oligonucleotides for these candidate targets. Antiseizure phenotypes were observed upon knockdown of miR-10a-5p, miR-21a-5p, and miR-142a-5p and electrophysiological analyses indicated broad safety of this approach. Combined inhibition of these three microRNAs reduced spontaneous seizures in epileptic mice. Proteomic data, RNA sequencing, and pathway analysis on predicted and validated targets of these microRNAs implicated derepressed TGF-ß signaling as a shared seizure-modifying mechanism. Correspondingly, inhibition of TGF-ß signaling occluded the antiseizure effects of the antagomirs. Together, these results identify shared, dysregulated, and functionally active microRNAs during the pathogenesis of epilepsy which represent therapeutic antiseizure targets.

[Case fatality rate-related factors in patients with bacteremia hospitalized due medical conditions in an institution of third level in Colombia, 2014-2016]. / Factores relacionados con letalidad en pacientes con bacteriemia hospitalizados por patología médica en una institución de tercer nivel en Colombia, 2014-2016.

BACKGROUND: Bloodstream infections are an increasing problem and currently represent a threat to public health, overcoming diseases such as HIV. Bacteremia accounts for approximately 15% of all nosocomial infections and affects 1% of all hospitalized patients. AIM: To describe the clinical, epidemiological and microbiological characteristic of episodes of nosocomial bacteremia occurring in a Colombian hospital. METHODS: Retrospective, observational, cross-sectional study including adult patients, hospitalized in the internal medicine unit at the University Hospital of Santander, Bucaramanga, Colombia, during years 2014 to 2016, who met the criteria of the CDC for bloodstream infection. The protocol was approved by the Hospital Ethics Committee and by the Research Ethics Committee of the Industrial University of Santander. RESULTS: We reviewed 450 clinical records with 148 patients and 182 microbiological isolates. 53% were male. The most frequent comorbidities were: high blood pressure (46.6%), HIV infection (29.7%). The vascular and urinary systems were the most frequent anatomical sites as the source of the infection (respectively 37.3% and 38.3%). Case fatality rate was 29%. The pathogens most frequently isolated were: Klebsiella pneumoniae, Acinetobacter baumannii, Escherichia coli, Pseudomonas aeruginosa (globally: 49.8%) and Staphylococcus aureus 12.1%. The multivariate analysis showed a relationship between anemia and in-hospital mortality (OR = 17.3, 95%CI 2.95-102.0). CONCLUSIONS: Bacteremia is a frequent infection during hospital care that presents high mortality. It is noteworthy the predominance of Enterobacteriaceae isolates with broad profiles of resistance. The history of HIV infection is one of the most frequent which deserves to be evaluated as a risk group.

Update on natural food pigments - A mini-review on carotenoids, anthocyanins, and betalains.

Extensive structure elucidation has revealed a remarkable diversity of structures for carotenoids, anthocyanins, and betalains, the major natural pigments in plant-derived foods. Composition, stability, influencing factors, processing effects have been widely investigated. Carotenoids isomerize and oxidize while anthocyanins undergo hydrolysis, nucleophilic attack of water, ring fission, and polymerization during thermal processing. Betacyanins suffer deglycosylation, isomerization, dehydrogenation, hydrolysis, and decarboxylation. Biotechnological production dominates research on carotenoids as food colorants while the search for plant sources continues with anthocyanins and betalains. Stabilization studies presently focus on microencapsulation and nanoencapsulation. For anthocyanins, co-pigmentation has also been intensely researched. Carotenoids have been the most studied in terms of health effects, involving epidemiological, cell, animal, and human intervention studies, yet some inconsistencies in the results persist. A wide range of biological activities have been attributed to anthocyanins and betalains, based mainly on cell and animal studies; human clinical studies are lacking.

Estabilidad de la hemoglobina sérica posterior a la transfusión de glóbulos rojos en pacientes adultos en el servicio de medicina interna / Stability of serum hemoglobin after red blood cell transfusion in adult patients assisted at an internal medicine service

Resumen OBJETIVO: Evaluar la estabilidad de la concentración de hemoglobina sérica posterior a la transfusión de glóbulos rojos empaquetados en el tiempo. MATERIAL Y MÉTODO: Estudio de cohorte prospectivo analítico, efectuado de septiembre de 2015 a mayo de 2016, que incluyó pacientes mayores de 18 años de edad, quienes cursaban con anemia que fue corregida mediante la transfusión de glóbulos rojos empaquetados. Se cuantificó la concentración de hemoglobina sérica inicial, una y seis horas después de la transfusión de glóbulos rojos empaquetados con el dispositivo HemoCue B Hemoglobin. Se evaluó la estabilidad de la concentración de hemoglobina sérica. Se consideró significativo un cambio en la concentración sérica de hemoglobina > 0.5 g/dL. RESULTADOS: Se incluyeron 121 pacientes. Los diagnósticos principales fueron: sepsis (60.3%), enfermedad renal crónica (31.4%) y cáncer hematológico (24.8%). La hemoglobina sérica promedio inicial fue de 6.9 ± 4.4 g/dL, después de la transfusión de glóbulos rojos empaquetados fue de 9.2 ± 1.5 g/dL (a la hora) y de 9.19 ± 1.5 g/dL (a las seis horas). La diferencia en la concentración de hemoglobina fue -0.007 g/dL (p = 0.94). Mediante un modelo de regresión logística se documentó la estabilidad de la concentración de hemoglobina sérica en el tiempo. CONCLUSIONES: La concentración de hemoglobina posterior a la transfusión de glóbulos rojos empaquetados es estable en el tiempo y no se ve afectada por los padecimientos concomitantes, número de unidades de glóbulos rojos empaquetados administradas y variables antropométricas.

Abstract OBJECTIVE: To evaluate the stability of serum hemoglobin concentration after packaged red blood cell units (PRBC) transfusion. MATERIAL AND METHOD: A prospective analytical cohort was done from September 2015 to May 2016 including patients older than 18 years who were enrolled with the diagnosis of anemia that was corrected by PRBC transfusion (n = 121). Initial serum hemoglobin concentration was quantified at one hour and six hours after PRBC transfusion with the HemoCue B Hemoglobin device. The stability of post-transfusion serum hemoglobin was evaluated. A change in serum hemoglobin concentration > 0.5 g/dL was considered significant. RESULTS: The main diagnoses were sepsis (60.3%), chronic kidney disease (31.4%) and hematologic cancer (24.8%). The initial mean serum hemoglobin was 6.9 ± 4.4 g/ dL, at one hour 9.2 ± 1.5 g/dL and 9.19 ± 1.5 g/dL at 6 hours following PRBC transfusion. The difference in hemoglobin concentration was -0.007 g/dL (p = 0.94). Using a logistic regression model the stability of serum hemoglobin concentration over time was documented. CONCLUSION: The hemoglobin concentration following PRBC transfusion is stable over time, and is not affected by concomitant diseases, number of PRBC units administered and anthropometric variables.

Context-Specific Switch from Anti- to Pro-epileptogenic Function of the P2Y1 Receptor in Experimental Epilepsy.

Extracellular ATP activates inflammatory responses to tissue injury. It is also implicated in establishing lasting network hyperexcitability in the brain by acting upon independent receptor systems. Whereas the fast-acting P2X channels have well-established roles driving neuroinflammation and increasing hyperexcitability, the slower-acting metabotropic P2Y receptors have received much less attention. Recent studies of P2Y1 receptor function in seizures and epilepsy have produced contradictory results, suggesting that the role of this receptor during seizure pathology may be highly sensitive to context. Here, by using male mice, we demonstrate that the metabotropic P2Y1 receptor mediates either proconvulsive or anticonvulsive responses, dependent on the time point of activation in relation to the induction of status epilepticus. P2Y1 deficiency or a P2Y1 antagonist (MRS2500) administered before a chemoconvulsant, exacerbates epileptiform activity, whereas a P2Y1 agonist (MRS2365) administered at this time point is anticonvulsant. When these drugs are administered after the onset of status epilepticus, however, their effect on seizure severity is reversed, with the antagonist now anticonvulsant and the agonist proconvulsant. This result was consistent across two different mouse models of status epilepticus (intra-amygdala kainic acid and intraperitoneal pilocarpine). Pharmacologic P2Y1 blockade during status epilepticus reduces also associated brain damage, delays the development of epilepsy and, when applied during epilepsy, suppresses spontaneous seizures, in mice. Our data show a context-specific role for P2Y1 during seizure pathology and demonstrate that blocking P2Y1 after status epilepticus and during epilepsy has potent anticonvulsive effects, suggesting that P2Y1 may be a novel candidate for the treatment of drug-refractory status epilepticus and epilepsy.SIGNIFICANCE STATEMENT This is the first study to fully characterize the contribution of a metabotropic purinergic P2Y receptor during acute seizures and epilepsy. The findings suggest that targeting P2Y1 may offer a potential novel treatment strategy for drug-refractory status epilepticus and epilepsy. Our data demonstrate a context-specific role of P2Y1 activation during seizures, switching from a proconvulsive to an anticonvulsive role depending on physiopathological context. Thus, our study provides a possible explanation for seemingly conflicting results obtained between studies of different brain diseases where P2Y1 targeting has been proposed as a potential treatment strategy and highlights that the timing of pharmacological interventions is of critical importance to the understanding of how receptors contribute to the generation of seizures and the development of epilepsy.