Letteratura scientifica selezionata sul tema "Sodium channels – Animal models"

In 2000, with the completion of the human genome project, nine related channels were found to comprise the complete voltage-gated sodium gene family and they were renamed NaV1.1–NaV1.9. This millennial event reflected the extraordinary impact of molecular genetics on our understanding of electrical signalling in the nervous system. In this review, studies of animal electricity from the time of Galvani to the present day are described. The seminal experiments and models of Hodgkin and Huxley coupled with the discovery of the structure of DNA, the genetic code and the application of molecular genetics have resulted in an appreciation of the extraordinary diversity of sodium channels and their surprisingly broad repertoire of functions. In the present era, unsuspected roles for sodium channels in a huge range of pathologies have become apparent.

Abstract Cardiomyocytes express a surprisingly large number of potassium channel types. The primary physiological functions of the currents conducted by these channels are to maintain the resting membrane potential and mediate action potential repolarization under basal conditions and in response to changes in the concentrations of intracellular sodium, calcium, and ATP/ADP. Here, we review the diversity and functional roles of cardiac potassium channels under normal conditions and how heritable mutations in the genes encoding these channels can lead to distinct arrhythmias. We briefly review atrial fibrillation and J-wave syndromes. For long and short QT syndromes, we describe their genetic basis, clinical manifestation, risk stratification, traditional and novel therapeutic approaches, as well as insights into disease mechanisms provided by animal and cellular models.

During epileptogenesis a series of molecular and cellular events occur, culminating in an increase in neuronal excitability, leading to seizure initiation. The entorhinal cortex has been implicated in the generation of epileptic seizures in both humans and animal models of temporal lobe epilepsy. This hyperexcitability is due, in part, to proexcitatory changes in ion channel activity. Sodium channels play an important role in controlling neuronal excitability, and alterations in their activity could facilitate seizure initiation. We sought to investigate whether medial entorhinal cortex (mEC) layer II neurons become hyperexcitable and display proexcitatory behavior of Na channels during epileptogenesis. Experiments were conducted 7 days after electrical induction of status epilepticus (SE), a time point during the latent period of epileptogenesis and before the onset of seizures. mEC layer II stellate neurons from post-SE animals were hyperexcitable, eliciting action potentials at higher frequencies compared with control neurons. Na channel currents recorded from post-SE neurons revealed increases in Na current amplitudes, particularly persistent and resurgent currents, as well as depolarized shifts in inactivation parameters. Immunocytochemical studies revealed increases in voltage-gated Na (Nav) 1.6 isoform levels. The toxin 4,9-anhydro-tetrodotoxin, which has greater selectivity for Nav1.6 over other Na channel isoforms, suppressed neuronal hyperexcitability, reduced macroscopic Na currents, persistent and resurgent Na current densities, and abolished depolarized shifts in inactivation parameters in post-SE neurons. These studies support a potential role for Nav1.6 in facilitating the hyperexcitability of mEC layer II neurons during epileptogenesis.

AbstractIntroductionThe pathophysiologic and neurochemical characteristics of neuropathic pain must be considered in the search for new treatment targets. Breakthroughs in the understanding of the structural and biochemical changes in neuropathy have opened up possibilities to explore new treatment paradigms. However, long term sequels from the damage are still difficult to treat.Aim of the studyTo examine the validity of pharmacological treatments in humans and animals for neuropathic pain.MethodAn overview from the literature and own experiences of pharmacological treatments employed to interfere in pain behavior in different animal models was performed.ResultsThe treatment principles tested in animal models of neuropathic pain may have predictive validity for treatment of human neuropathies. Opioids, neurotransmitter blockers, drugs interfering with the prostaglandin syntheses as well as voltage gated sodium channel blockers and calcium channel blockers are treatment principles having efficacy and similar potency in humans and in animals. Alternative targets have been identified and have shown promising results in the validated animal models. Modulators of the glutamate system with an increased expression of glutamate re-uptake transporters, inhibition of pain promoters as nitric oxide and prostaglandins need further exploration. Modulation of cytokines and neurotrophins in neuropathic pain implies new targets for study. Further, a combination of different analgesic treatments may as well improve management of neuropathic pain, changing the benefit/risk ratio.ImplicationsNot surprisingly most pharmacologic principles that are tested in animal models of neuropathic pain are also found to be active in humans. Whereas many candidate drugs that were promising in animal models of neuropathic pain turned out not to be effective or too toxic in humans, animal models for neuropathic pain are still the best tools available to learn more about mechanisms of neuropathic pain. Better understanding of pathogenesis is the most hopeful approach to improve treatment of neuropathic pain.

Abstract Chronic cough is a significant clinical problem in many patients. Current cough suppressant therapies are largely ineffective and have many dangerous adverse effects. Therefore, the identification of novel therapeutic targets and strategies for chronic cough treatment may lead to development of novel effective antitussive therapies with fewer adverse effects. The experimental research in the area of airway sensory nerves suggests that there are two main vagal afferent nerve subtypes that can directly activate cough – extrapulmonary airway C-fibres and Aδ-fibres (described as cough receptors) innervating the trachea. There are different receptors on the vagal nerve terminals that can trigger coughing, such as TRP channels and P2X2/3 receptors. However, in many patients with chronic respiratory diseases multiple activation of these receptors could be involved and it is also difficult to target these receptors. For that reason, a strategy that would inhibit cough-triggering nerve afferents regardless of activated receptors would be of great benefit. In recent years huge progress in understanding of voltage-gated sodium channels (NaVs) leads to a hypothesis that selective targeting of NaVs in airways may represent an effective treatment of pathological cough. The NaVs (NaV1.1 – NaV1.9) are essential for initiation and conduction of action potentials in these nerve fibres. Effective blocking of NaVs will prevent communication between airways and central nervous system and that would inhibit provoked cough irrespective to stimuli. This review provides an overview of airway afferent nerve subtypes that have been described in respiratory tract of human and in animal models. Moreover, the review highlights the current knowledge about cough, the sensory nerves involved in cough, and the voltage-gated sodium channels as a novel neural target in regulation of cough.

Basic research using animal models points to a causal role of the central nervous system in essential hypertension; however, since clinical research is technically difficult to perform, this connection has not been confirmed in humans. Recently, renal nerve ablation in humans proved to continuously decrease blood pressure in resistant hypertension. Furthermore, when electrical stimulation was continuously applied to the carotid baroreceptor nerve of human adults, their blood pressure lowered. These findings promoted the concept that the central nervous system may actually be involved in the pathogenesis of essential hypertension, which is closely associated with excess sodium intake. We have demonstrated that endogenous digitalis plays a key role in hypertension associated with excess sodium intake via sympathetic activation in rats. Increased sodium concentration inside the brain activates epithelial sodium channels and the renin-angiotensin-aldosterone system in the brain. Aldosterone releases ouabain from neurons in the paraventricular nucleus in the hypothalamus. Angiotensin II and aldosterone of peripheral origin reach the brain to augment sympathetic outflow. Collectively essential hypertension associated with excess sodium intake and obesity, renovascular hypertension, and primary aldosteronism and pseudoaldosteronism all seem to have a common cause originating from the central nervous system.

Insulin has been shown to have antinatriuretic actions in humans and animal models. Moreover, endogenous hyperinsulinemia and insulin infusion have been correlated to increased blood pressure in some models. In this review, we present the current state of understanding with regard to the regulation of the major renal sodium transporters by insulin in the kidney. Several groups, using primarily cell culture, have demonstrated that insulin can directly increase activity of the epithelial sodium channel, the sodium-phosphate cotransporter, the sodium-hydrogen exchanger type III, and Na-K-ATPase. We and others have demonstrated alterations in the expression at the protein level of many of these same proteins with insulin infusion or in hyperinsulinemic models. We also discuss how this regulation is perturbed in type I and type II diabetes mellitus. Finally, we discuss a potential role for regulation of insulin receptor signaling in the kidney in contributing to sodium balance and blood pressure.

Disruption of cardiac electrical activity resulting in palpitations and syncope is often an early symptom of pregnancy. Pregnancy is a time of dramatic and dynamic physiological and hormonal changes during which numerous demands are placed on the heart. These changes result in electrical remodelling which can be detected as changes in the electrocardiogram (ECG). This gestational remodelling is a very under-researched area. There are no systematic large studies powered to determine changes in the ECG from pre-pregnancy, through gestation, and into the postpartum period. The large variability between patients and the dynamic nature of pregnancy hampers interpretation of smaller studies, but some facts are consistent. Gestational cardiac hypertrophy and a physical shift of the heart contribute to changes in the ECG. There are also electrical changes such as an increased heart rate and lengthening of the QT interval. There is an increased susceptibility to arrhythmias during pregnancy and the postpartum period. Some changes in the ECG are clearly the result of changes in ion channel expression and behaviour, but little is known about the ionic basis for this electrical remodelling. Most information comes from animal models, and implicates changes in the delayed-rectifier channels. However, it is likely that there are additional roles for sodium channels as well as changes in calcium homoeostasis. The changes in the electrical profile of the heart during pregnancy and the postpartum period have clear implications for the safety of pregnant women, but the field remains relatively undeveloped.

The present study aimed to design and synthesize a new series of hybrid compounds with pyrrolidine-2,5-dione and thiophene rings in the structure as potential anticonvulsant and antinociceptive agents. For this purpose, we obtained a series of new compounds and evaluated their anticonvulsant activity in animal models of epilepsy (maximal electroshock (MES), psychomotor (6 Hz), and subcutaneous pentylenetetrazole (scPTZ) seizure tests). To determine the mechanism of action of the most active anticonvulsant compounds (3, 4, 6, 9), their influence on the voltage-gated sodium and calcium channels as well as GABA transporter (GAT) was assessed. The most promising compound 3-(3-methylthiophen-2-yl)-1-(3-morpholinopropyl)pyrrolidine-2,5-dione hydrochloride (4) showed higher ED50 value than those of the reference drugs: valproic acid (VPA) and ethosuximide (ETX) (62.14 mg/kg vs. 252.7 mg/kg (VPA) in the MES test, and 75.59 mg/kg vs. 130.6 mg/kg (VPA) and 221.7 mg/kg (ETX) in the 6 Hz test, respectively). Moreover, in vitro studies of compound 4 showed moderate but balanced inhibition of the neuronal voltage-sensitive sodium (site 2) and L-type calcium channels. Additionally, the antinociceptive activity of the most active compounds (3, 4, 6, 9) was also evaluated in the hot plate test and writhing tests, and their hepatotoxic properties in HepG2 cells were also investigated. To determine the possible mechanism of the analgesic effect of compounds 3, 6, and 9, the affinity for the TRPV1 receptor was investigated.

Cerebral ischaemia is a leading cause of disability and death world-wide. The only effective treatments are thrombolytic therapy (plasminogen activator; tPA) and hypothermia (33?C). However, tPA has limited clinical application due to its short therapeutic time window and its specific application in thrombo-embolic stroke. Moderate hypothermia (33?C) is only being used following cardiac arrest in comatose survivors. Hence more treatments are urgently required. The first step in developing new treatments is the identification and characterisation of a potential therapeutic target. Since brain damage following cerebral ischaemia is associated with disturbances in intracellular calcium homeostasis, the sodium-calcium exchanger (NCX) is a potential therapeutic target due to its ability to regulate intracellular calcium. Currently, however there is uncertainty as to whether the plasma membrane NCX has a neuroprotective or neurodamaging role following cerebral ischemia. To address this issue I compared hippocampal neuronal injury in NCX3 knockout mice (Ncx3-/-) and wild-type mice (Ncx3+/+) following global cerebral ischaemia. In order to perform this study I first established a bilateral common carotid occlusion (BCCAO) model of global ischaemia in wild-type C57/BlHsnD mice using controlled ventilation. After trials of several ischaemic time points, 17 minutes was established as the optimum duration of ischaemia to produce selective hippocampal CA1 neuronal loss in the wild-type mice. I then subjected NCX3 knockout and wild-type mice to 17 minutes of ischaemia. Following the 17 minute period of ischaemia, wild-type mice exhibited 80% CA1 neuronal loss and 40% CA2 neuronal loss. In contrast, NCX3 knockout mice displayed > 95% CA1 neuronal loss and 95% CA2 neuronal loss. Following experiments using a 17 minute duration of global ischaemia, a 15 minute duration of ischaemia was also evaluated. Wild-type mice exposed to a 15 minute period of ischaemia, did not exhibit any significant hippocampal neuronal loss. In contrast, NCX3 knockout mice displayed 45% CA1 neuronal loss and 25% CA2 neuronal loss. The results clearly demonstrate that mice deficient for the NCX3 protein are more susceptible to global cerebral ischaemia than wild-type mice. My findings showing a neuroprotective role for NCX3 following ischaemia, suggest that the exchanger has a positive role in maintaining neuronal intracellular calcium homeostasis. When this function is disrupted, neurons are more susceptible to calcium deregulation, with resultant cell death via calcium mediated pathways. Therefore, improving NCX activity following cerebral ischaemia may provide a therapeutic strategy to reduce neuronal death.

Epilepsy accounts for 0.5% of the global burden of disease, and primary prevention of epilepsy represents one of the three 2007 NINDS Epilepsy Research Benchmarks. Efforts to understand and intervene in the process of epileptogenesis have yielded fruitful preventative strategies in animal models. This article reviews the current understanding of epileptogenesis, introduces the concept of a "critical period" for epileptogenesis, and examines strategies for epilepsy prevention in animal models of both acquired and genetic epilepsies. As proof of principle, we investigated whether early preventative treatment during epileptogenesis in the WAG/Rij rat model of primary generalized epilepsy would persistently suppress the epilepsy phenotype in adulthood. Oral ethosuximide was given from age p21 to 5 months, covering the established period for epileptogenesis in this model. We then assessed the epilepsy phenotype by performing electroencephpalogram (EEG) recordings at serial time points after treatment cessation and by immunocytochemically measuring the cortical expression of ion channels Nav1.1, Nav1.6, and HCN1, which are dysregulated in epileptic WAG/Rij rats. Treatment both persistently suppressed seizures, even up to 3 months after treatment cessation, and blocked ion channel dysregulation. These findings indicated that treatment during epileptogenesis prevented the development of the epileptic phenotype. Subsequently, we investigated the C3H/HeJ mouse model of genetic epilepsy as a candidate for future studies in preventative treatment during epileptogenesis. Serial EEG recordings were performed from p5 to 3 months of age. We found that C3H/HeJ mice underwent three distinct, stereotyped phases of seizure development, which suggests that this model would be an appropriate candidate for future research on prevention of epileptogenesis.

Thesis (M.S.)--University of Missouri-Columbia, 2005.
The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Vita. "May 2005" Includes bibliographical references.

Le syndrome de l'intestin irritable (SII) touche près de 10% de la population. Nous avons utilisé un modèle animal de SII induit par le butyrate développé au laboratoire afin de décortiquer les mécanismes de l'hypersensibilité colique (HSC) dans le SII. Le blocage du NGF par des anticorps anti-NGF prévient l'HSC induite par le butyrate, évaluée par le test de distension colorectale. Le NGF, quantifié par immunohistochimie (IHC), est surexprimé dans les ganglions rachidiens dorsaux (GRD) innervant le côlon des rats butyrate. Le blocage des canaux ASIC par amiloride prévient l'HSC induite par le butyrate. L'expression des ARNm ASIC1a et ASIC1b, évaluée par RT-PCR, est augmentée dans les GDR des rats butyrate. Cette augmentation est corrélée à une augmentation de l'expression de la protéine ASIC1A dans les neurones nociceptifs, quantifiée par IHC. Le blocage du NGF par des anticorps anti-NGF prévient la surexpression de ASIC1A dans les GRD. L'absence de variation d'expression du NGF et de ASIC1A au niveau colique suggère que ces moléciles ont une implication dans l'élément présynaptique plutôt que dans les terminaisons libres coliques. L'étude de l'expression spinale de la protéine Fos après stimulation des fibres coliques a montré que l'HSC induite par le butyrate est associée à une activation spécifique des segments thoraciques T10-T11-T12 de la moëlle épinière (MEp). Le blocage spinal du canal ASIC1A par la PcTx1 prévient l'HSC induite par le butyrate. L'expression de ASIC1a, évaluée par RT-PCR et Western blot, est augmentée dans la MEp des rats butyrate. Comme à la périphérie, l'expression de ASIC1a est modulée par le NGF puisque le blocage du NGF prévient la surexpression des ARNm et de la protéine ASIC1A dans la MEp des rats butyrate. En conclusion, ce travail de thèse suggère que le NGF et le canal ASIC1A jouent un rôle critique dans le développement de douleurs viscérales en contribuant à la fois à la sensibilisation périphérique et centrale.

[ES] Las enfermedades cardiovasculares siguen siendo la principal causa de muerte en Europa. Las arritmias cardíacas son una causa importante de muerte súbita, pero sus mecanismos son complejos. Esto denota la importancia de su estudio y prevención. La investigación sobre electrofisiología cardíaca ha demostrado que las anomalías eléctricas causadas por mutaciones que afectan a canales cardíacos pueden desencadenar arritmias. Sorprendentemente, se ha descubierto una gran variedad de fármacos proarrítmicos, incluidos aquellos que usamos para prevenirlas. Las indicaciones de uso de fármacos actuales intentaron solucionar este problema diseñando una prueba para identificar aquellos fármacos que podían ser peligrosos basado en el bloqueo de un solo canal iónico. El estudio de las interacciones fármaco-canal ha revelado la existencia no sólo de compuestos que bloquean múltiples canales, sino también una gran complejidad en esas interacciones. Esto podría explicar por qué algunos medicamentos pueden mostrar efectos muy diferentes en la misma enfermedad. Existen dos desafíos importantes con respecto a los efectos de los fármacos en la electrofisiología cardíaca. Por un lado, las empresas y entidades reguladoras están buscando una herramienta de alto rendimiento que mejore la detección del potencial proarrítmico durante el desarrollo de fármacos. Por otro lado, los pacientes con anomalías eléctricas a menudo requieren tratamientos personalizados más seguros. Las simulaciones computacionales contienen un poder sin precedentes para abordar fenómenos biofísicos complejos. Deberían ser de utilidad a la hora de determinar las características que definen tanto los efectos beneficiosos como no deseados de los fármacos mediante la reproducción de datos experimentales y clínicos. En esta tesis doctoral, se han utilizado modelos computacionales y simulaciones para dar respuesta a estos dos desafíos. El estudio de los efectos de los fármacos sobre la actividad cardíaca se dividió en el estudio de su seguridad y de su eficacia, respectivamente. Para dar respuesta al primer desafío, se adoptó un enfoque más amplio y se generó un nuevo biomarcador fácil de usar para la clasificación del potencial proarrítmico de los fármacos utilizando modelos del potencial de acción de células y tejidos cardíacos humanos. Se integró el bloqueo de múltiples canales a través de IC50 y el uso de concentraciones terapéuticas con el fin de mejorar el poder predictivo. Luego, se entrenó el biomarcador cuantificando el potencial proarrítmico de 84 fármacos. Los resultados obtenidos sugieren que el biomarcador podría usarse para probar el potencial proarrítmico de nuevos fármacos. Respecto al segundo desafío, se adoptó un enfoque más específico y se buscó mejorar la terapia de pacientes con anomalías eléctricas cardíacas. Por lo tanto, se creó un modelo detallado de la mutación V411M del canal de sodio, causante del síndrome de QT largo, reproduciendo datos clínicos y experimentales. Se evaluaron los posibles efectos beneficiosos de ranolazina, a la par que se aportó información sobre los mecanismos que impulsan la efectividad de la flecainida. Los resultados obtenidos sugieren que, si bien ambos fármacos mostraron diferentes mecanismos de bloqueo de los canales de sodio, un tratamiento con ranolazina podría ser beneficioso en estos pacientes.
[CA] Les malalties cardiovasculars continuen sent la principal causa de mort a Europa. Les arrítmies cardíaques són una causa important de mort sobtada, però els seus mecanismes són complexos. Això denota la importància del seu estudi i prevenció. La investigació sobre electrofisiologia cardíaca ha demostrat que les anomalies elèctriques que afecten a canals cardiacs poden desencadenar arrítmies. Sorprenentment, s'ha descobert una gran varietat de fàrmacs proarrítmics, inclosos aquells que utilitzem per a previndre-les. Les indicacions d'ús de fàrmacs actuals van intentar solucionar aquest problema dissenyant una prova per a identificar aquells fàrmacs que podien ser perillosos basada en el bloqueig d'un sol canal iònic. L'estudi de les interaccions fàrmac-canal ha revelat l'existència no sols de compostos que bloquegen múltiples canals, sinó també una gran complexitat en aquestes interaccions. Això podria explicar per què alguns medicaments poden mostrar efectes molt diferents en la mateixa malaltia. Existeixen dos desafiaments importants respecte als efectes dels fàrmacs en la electrofisiologia cardíaca. D'una banda, les empreses i entitats reguladores estan buscant una eina d'alt rendiment que millore la detecció del potencial proarrítmic durant el desenvolupament de fàrmacs. D'altra banda, els pacients amb anomalies elèctriques sovint requereixen tractaments personalitzats més segurs. Les simulacions computacionals contenen un poder sense precedents per a abordar fenòmens biofísics complexos. Haurien de ser d'utilitat a l'hora de determinar les característiques que defineixen tant els efectes beneficiosos com no desitjats dels fàrmacs mitjançant la reproducció de dades experimentals i clíniques. En aquesta tesi doctoral, s'han utilitzat models computacionals i simulacions per a donar resposta a aquests dos desafiaments. L'estudi dels efectes dels fàrmacs sobre l'activitat cardíaca es va dividir en l'estudi de la seva seguretat i la seva eficacia. Per a donar resposta al primer desafiament, es va adoptar un enfocament més ampli i es va generar un nou biomarcador fàcil d'usar per a la classificació del potencial proarrítmic dels fàrmacs utilitzant models del potencial d'acció de cèl·lules i teixits cardíacs humans. Es va integrar el bloqueig de múltiples canals a través d'IC50 i l'ús de concentracions terapèutiques amb la finalitat de millorar el poder predictiu. Després, es va entrenar el biomarcador quantificant el potencial proarrítmic de 84 fàrmacs. Els resultats obtinguts suggereixen que el biomarcador podria usar-se per a provar el potencial proarrítmic de nous fàrmacs. Respecte al segon desafiament, es va adoptar un enfocament més específic i es va buscar millorar la teràpia de pacients amb anomalies elèctriques cardíaques. Per tant, es va crear un model detallat de la mutació V411M del canal de sodi, causant de la síndrome de QT llarg, reproduint dades clíniques i experimentals. Es van avaluar els possibles efectes beneficiosos de ranolazina, a l'una que es va aportar informació sobre els mecanismes que impulsen l'efectivitat de la flecainida. Els resultats obtinguts suggereixen que, si bé tots dos fàrmacs van mostrar diferents mecanismes de bloqueig dels canals de sodi, un tractament amb ranolazina podria ser beneficiós en aquests pacients.
[EN] Cardiovascular disease remains the main cause of death in Europe. Cardiac arrhythmias are an important cause of sudden death, but their mechanisms are complex. This denotes the importance of their study and prevention. Research on cardiac electrophysiology has shown that electrical abnormalities caused by mutations in cardiac channels can trigger arrhythmias. Surprisingly, a wide variety of drugs have also shown proarrhythmic potential, including those that we use to prevent arrhythmia. Current guidelines designed a test to identify dangerous drugs by assessing their blocking power on a single ion channel to address this situation. Study of drug-channel interactions has revealed not only compounds that block multiple channels but also a great complexity in those interactions. This could explain why similar drugs can show vastly different effects in some diseases. There are two important challenges regarding the effects of drugs on cardiac electrophysiology. On the one hand, companies and regulators are in search of a high throughput tool that improves proarrhythmic potential detection during drug development. On the other hand, patients with electrical abnormalities often require safer personalized treatments owing to their condition. Computer simulations provide an unprecedented power to tackle complex biophysical phenomena. They should prove useful determining the characteristics that define the drugs' beneficial and unwanted effects by reproducing experimental and clinical observations. In this PhD thesis, we used computational models and simulations to address the two abovementioned challenges. We split the study of drug effects on the cardiac activity into the study of their safety and efficacy, respectively. For the former, we took a wider approach and generated a new easy-to-use biomarker for proarrhythmic potential classification using cardiac cell and tissue human action potential models. We integrated multiple channel block through IC50s and therapeutic concentrations to improve its predictive power. Then, we quantified the proarrhythmic potential of 84 drugs to train the biomarker. Our results suggest that it could be used to test the proarrhythmic potential of new drugs. For the second challenge, we took a more specific approach and sought to improve the therapy of patients with cardiac electrical abnormalities. Therefore, we created a detailed model for the long QT syndrome-causing V411M mutation of the sodium channel reproducing clinical and experimental data. We tested the potential benefits of ranolazine, while giving insights into the mechanisms that drive flecainide's effectiveness. Our results suggest that while both drugs showed different mechanisms of sodium channel block, ranolazine could prove beneficial in these patients.
This PhD thesis was developed within the following projects: Ministerio de Economía y Competitividad and Fondo Europeo de Desarrollo Regional (FEDER) DPI2015-69125-R (MINECO/FEDER, UE): Simulación computacional para la predicción personalizada de los efectos de los fármacos sobre la actividad cardiaca. Dirección General de Política Científica de la Generalitat Valenciana (PROMETEU2016/088): “Modelos computacionales personalizados multiescala para la optimización del diagnóstico y tratamiento de arritmias cardiacas (personalised digital heart). Vicerrectorado de Investigación, Innovación y Transferencia de la Universitat Politècnica de València, Ayuda a Primeros Proyectos de Investigación (PAID-06-18), and by Memorial Nacho Barberá. Instituto de Salud Carlos III (La Fe Biobank PT17/0015/0043).
Cano García, J. (2021). Prediction of the effects of drugs on cardiac activity using computer simulations [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/164094
TESIS

As doenças cardiovasculares são a maior causa de morte no mundo e entre essas doenças, a hipertrofia cardíaca (HC) tem se destacado especialmente por ser um fator de risco de insuficiência cardíaca. A HC é um fenômeno que acompanha a hipertensão arterial e no qual se observa aumento de proteínas estruturais e contráteis dos cardiomiócitos, havendo muitas vezes concomitantemente aumento do colágeno intersticial. Fatores independentes da pressão arterial também podem contribuir para o desenvolvimento da hipertrofia cardíaca. Dentre estes fatores, a sobrecarga de sal na dieta tem se destacado. Diversos estudos comprovam o efeito hipertrófico do sal. Em modelos animais onde se estudou sobrecarga de sal, não foi detectado aumento da atividade de renina plasmática, sugerindo que o sistema renina-angiotensina aldosterona (SRA) circulante pode não estar envolvido no desenvolvimento da hipertrofia cardíaca. Apesar de alguns estudos tentarem elucidar o papel do sal no desenvolvimento da hipertrofia ventricular esquerda, os mecanismos pelo qual o sal atua ainda não estão totalmente esclarecidos. Neste contexto, o objetivo do presente estudo é observar os fenômenos que ocorrem no ventrículo esquerdo em resposta a sobrecarga de sal na dieta na tentativa de elucidar sua fisiopatologia. Para tanto, ratos Wistar machos foram divididos em cinco grupos de acordo com a dieta (normossódica 1,26% e hipersódica 8% de NaCl) e com o tratamento (losartan, cloridrato de hidralazina ou N-acetilcisteína). Foi avaliada a evolução ponderal, pressão arterial caudal, medida do diâmetro transverso do cardiomiócito, fibrose intersticial, expressão gênica e proteica dos componentes do SRA, dosagem de aldosterona sérica e cardíaca, dosagem de TBARS cardíaco, concentração de angiotensina II e estado conformacional dos receptores AT1 e AT2. Os principais resultados observados foram: o aumento do consumo de ração (com elevada concentração de NaCl) do grupo HS+NAC e consequente aumento na pressão arterial e peso corpóreo; o desenvolvimento de HC independente do incremento da pressão arterial no grupo HS+HZ e a prevenção total ou parcial dessa hipertrofia através dos tratamentos com losartan e N-acetilcisteína, respectivamente e prevenção da fibrose intersticial nos grupos tratados com hidralazina, losartan e N-acetilcisteína
Cardiovascular diseases are the leading cause of death worldwide and among these diseases, the cardiac hypertrophy (CH) has been highlighted, especially as an important risk factor for developing heart failure. The CH is a phenomenon that accompanies hypertension and in which there is increased structural and contractile proteins in cardiomyocytes, with often concomitant increase of interstitial collagen. Blood pressure independent risk factors can also contribute to the development of cardiac hypertrophy. Among these factors, the high salt intake has been outstanding. Several studies confirm the hypertrophic effect of salt. In animal models submitted to salt overload, no increase in plasma renin activity was observed, suggesting that the renin-angiotensin (RAS) circulating system may not be involved in the development of cardiac hypertrophy. Although some studies attempting to elucidate the role of salt in the development of left ventricular hypertrophy, the mechanisms by which salt acts are not yet fully understood. In this context, the objective of this study is to observe the phenomena occurring in the left ventricle in response to dietary salt overload in an attempt to elucidate its pathophysiology.Male Wistar rats were divided into five groups according to their diet (1.26% and 8% NaCl) and treatment (losartan, hydralazine or N-acetylcysteine). We evaluated the body weight, tail-cuff blood pressure, the transverse diameter of the cardiomyocyte, interstitial fibrosis, gene and protein expression of RAAS components, serum and cardiac aldosterone dosage, cardiac TBARS, angiotensin II concentration and binding of conformation-specific anti-AT1 and anti-AT2 antibodies. The main results were: increased food intake (with high NaCl content) in the HS + NAC group and consequent increase in blood pressure and body weight; developing blood pressure-independent CH in the HS + HZ group partial or total prevention of such hypertrophy by treatment with losartan and N-acetylcysteine, respectively, and prevention of interstitial fibrosis in groups treated with hydralazine, losartan and N-acetylcysteine

Introdução: Diversos estudos indicaram consequências de alterações na nutrição materna durante a gestação sobre a saúde da prole adulta, tais como: hipertensão, doenças cardiovasculares, resistência à insulina, diabete melito e doença renal. No entanto, a literatura é pobre em avaliações decorrentes de modificações nutricionais maternas sobre a prole logo após o nascimento. Métodos: Ratas Wistar durante o período gestacional foram alimentadas com dieta hipossódica (HO - 0,15% de NaCl), normossódica (NR - 1,3% de NaCl) ou hipersódica (HR - 8% de Na Cl). Após o nascimento, nas primeiras vinte e quatro horas foram coletados rins e coração dos neonatos machos e fêmeas (n=6- 8/grupo) para verificar as possíveis alterações na estrutura cardíaca e renal pelo método de estereologia. Também foi avaliada a expressão proteica e gênica dos componentes do sistema renina angiotensina (SRA) no coração e rins através do método ELISA indireto e RT-qPCR. Resultados: O peso ao nascimento foi menor em machos e fêmeas da prole de mães alimentadas com dieta hipossódica durante a gestação quando comparado NR e HR. Não houve diferença no volume renal, volume de seus compartimentos (córtex, medula e pelve) e número de glomérulos entre os grupos experimentais (HO, NR e HR). No entanto, o número de glomérulos foi maior em fêmeas comparado aos machos nos três grupos experimentais. O diâmetro transverso do núcleo dos cardiomiócitos no ventrículo esquerdo e no ventrículo direito de machos da prole HR foi maior do que na prole NR. A expressão proteica do receptor AT1 no rim de machos da prole foi menor no grupo HO do que no grupo NR e HR. A expressão proteica do receptor AT2 também foi menor em machos do grupo HO do que no grupo NR. Não houve diferença entre os grupos na expressão proteica dos receptores AT1 e AT2 no rim das fêmeas. Conclusão: O presente estudo detectou alterações na estrutura cardíaca de neonatos machos, mas não em neonatos fêmeas decorrentes de sobrecarga de sal durante a gravidez. As alterações observadas na expressão dos receptores AT1 e AT2 no rim de neonatos machos podem ser responsáveis por alterações na função renal
Introduction: Several studies have shown several consequences on adult offspring due to alterations in maternal nutrition during pregnancy, such as: hypertension, heart diseases, insulin resistance, diabetes mellitus and kidney diseases. Nevertheless, few studies evaluated maternal nutritional alterations in neonates. Methods: Female Wistar rats where fed from day one of pregnancy until delivery with low- (LS - 0.15% NaCl), normal- (NS - 1.3% NaCl) or high- (HS - 8%NaCl) salt diet. During the first twenty-four hours after birth, newborn\'s (n=6- 8/group) kidneys and heart were collected to evaluate possible changes of their structure by stereology. The protein and the gene expression of the renin angiotensin system components were evaluated by indirect ELISA and by RTqPCR, respectively. Results: Birth weight was lower in male and female offspring of dams fed LS during pregnancy. No differences between groups (LS, NS and HS) were observed in total renal volume and its compartments (cortex, medulla and pelvis) and number of glomeruli. The number of glomeruli was higher in female when compared to male newborns in the three experimental groups. The transverse diameter of the nuclei of the cardiomyocytes was higher in HS in both left and right ventricle vs. NS. The AT1 receptor protein expression was lower in kidneys of LS than in NS and HS male newborns. AT2 receptor protein expression was also lower in male LS than in NS. No differences in AT1 and AT2 receptors protein expression in female newborn\'s kidneys were found. Conclusion: The present study shows changes in cardiac structure male but not of female neonates induced salt overload during pregnancy. The alterations observed in AT1 and AT2 expression in kidneys of neonates may be responsible for alteration in renal function

Tese de mestrado, Bioquímica, Universidade de Lisboa, Faculdade de Ciências, 2015
A capacidade de um indivíduo percecionar estímulos dolorosos é de extrema importância. Ao sentir ou associar uma situação a uma experiência dolorosa, o indivíduo pode adotar posturas protetoras, de forma a evitar possíveis danos causados pela situação eminente e/ou futuras situações semelhantes. Os mecanismos moleculares inerentes à deteção de estímulos (mecânicos, térmicos ou químicos) e capazes de codificar dor pelos neurónios sensoriais primários (nocicetores) integram o fenómeno de nociceção. Contudo, em situações fisiopatológicas, é possível que os doentes desenvolvam Dor crónica (DC), uma síndrome em que a perceção da dor é exacerbada e os doentes sofrem de hiperalgesia (reação exagerada a um estímulo doloroso), alodinia (sensação de dor a um estímulo não doloroso), dor espontânea, parestesia (sensação anormal) e disestesia (parestesia desconfortável). A DC afeta mais de 20% da população Humana e as suas etiologias são variadas: trauma, doenças autoimunes e metabólicas, infeções, tratamento antirretroviral e quimioterapia. Apesar da grande incidência, atualmente não existe uma abordagem terapêutica eficiente que possa suplantar as necessidades dos doentes, o que agrava as consequências psicológicas, sociais e económicas desta patologia. Os atuais tratamentos farmacológicos variam entre anti-inflamatórios não esteróides, opióides, antidepressivos e anticonvulsivos que, para além de frequentemente não serem eficientes, estão associados a efeitos secundários indesejáveis como a dependência, habituação ou até mesmo a perda da capacidade de gestão da dor. As fibras nervosas C e Aδ transmitem eletricamente os estímulos sensoriais, a velocidade de transmissão nas fibras C (0,5-2m-1∙s), devido à ausência de um revestimento de mielina, é consideravelmente mais lenta, comparando à velocidade de transmissão das fibras Aδ (6-30 m-1∙s). A perceção dos estímulos transmitidos pelas fibras C é difusa, ao passo que os transmitidos pelas fibras Aδ são percecionados como bem localizados. Todas as fibras nocicetivas são constituídas pelos neurónios do gânglio da raiz dorsal (DRG, do inglês Dorsal Root Ganglia). Estes últimos são classificados de acordo com o seu diâmetro em pequeno (27-31μm), médio (31-40μm) e grande (40-50μm); e possuem uma anatomia pseudounipolar, com um longo axónio que inerva a periferia do organismo e um axónio mais curto que se dirige para o corno da raiz dorsal da medula espinal e o corpo celular alojado em locais específicos no interior da coluna vertebral. Os conjuntos dos corpos celulares destes neurónios é então designado de gânglio da raiz dorsal e a sua localização dentro da coluna vertebral é específica às fibras aferentes correspondentes. Dado o foco da presente dissertação, será dada uma especial atenção aos neurónios-DRG de pequeno e médio diâmetro integrantes da nociceção, que compõem as fibras C e Aδ, respetivamente. Em situações fisiológicas, na ausência de estímulos capazes de produzir dor, os nocicetores encontram-se quiescentes, sem qualquer informação a ser transmitida. No entanto, na presença destes estímulos, os recetores nas terminações nervosas dos neurónios primários podem ser ativados. Quando o estímulo tem uma magnitude maior do que o limiar de ativação das fibras nervosas, é produzido um potencial de ação, que percorre a fibra nervosa, terminando nas lâminas I e II no interior do corno da raiz dorsal da medula espinal. A partir daí, a informação é transmitida para os neurónios secundários, para de seguida ser transmitida ao sistema nervoso central, onde no córtex sensorial do cérebro, a informação a relativa à quantificação e localização da dor é integrada. A geração e transmissão de potenciais de ação é particularmente devida à ativação dos canais de sódio (Na+) ativados pela voltagem (VGSC, do inglês voltage-gated sodium channels) presentes ao longo de todo o neurónio-DRG. Até à data estão identificadas nove isoformas diferentes de VGSC (Nav1.1-9), que diferem nas propriedades eletrofisiológicas e distribuição corporal. As isoformas Nav1.7, Nav1.8 e Nav1.9 encontram-se preferencialmente distribuídas nos neurónios-DRG responsáveis pela transmissão nocicetiva. Patologias relacionadas com a perda ou o ganho de função dos genes que codificam para o Nav1.7 e Nav1.8 estão associadas a anomalias na perceção de dor, nomeadamente hipo ou hipersensibilidade. Ainda existe muito por se desvendar na fisiopatologia da DC, no entanto, sabe-se que os neurónios-DRG se encontram hiperexcitáveis neste contexto, comparativamente a situações fisiológicas. Estudos anteriores têm evidenciado alterações nos canais iónicos ativados pela voltagem, nomeadamente VGSC, que levam a esse estado de hiperexcitabilidade nestes neurónios. A estratégica terapêutica alternativa mais promissora para tratar DC visa a redução dessa “hiperexcitabilidade neural” que pode ocorrer através da alteração do funcionamento de canais-iónicos-chave que modulam a atividade elétrica desses neurónios. O conhecimento sobre as correntes iónicas subjacentes ao funcionamento de determinados canais é assim preponderante. Os canais/correntes especificamente implicados na DC carecem ainda de um conhecimento mais aprofundado e da sua validação como potenciais alvos terapêuticos. A presente dissertação objetiva o estudo das características eletrofisiológicas das correntes de Na+, com direta relevância para a DC em neurónios-DRG de pequeno diâmetro, através do estudo da biofísica e expressão destes canais. Para tal, e uma vez que a DC pode ter várias etiologias, esta síndrome foi estudada no contexto de origem inflamatória e neuropática, baseando-se em modelos animais de dor em ratos. A DC inflamatória foi estudada através da indução de monoartrite no joelho do rato (ipsilateral), pela injeção de uma substância que ativa o sistema imunitário: Complete Freund’s Adjuvant (CFA). No caso da dor neuropática, usou-se o muito comum modelo Chronic Constriction Injury (CCI), que assenta na constrição crónica do nervo isquiático do rato (ipsilateral), ao nível da coxa, através de quatro nós soltos em torno do mesmo realizados cirurgicamente. O desenvolvimento de DC em ambos os modelos foi acompanhado ao longo do tempo através de testes comportamentais. Estes testes permitiram a observação do aparecimento de posturas putativas de evidenciarem dor espontânea em ratos. Foi também realizado o estudo do desenvolvimento de hiperalgesia, através da observação de reflexos protetores quando os ratos eram submetidos a estímulos mecânicos aplicados na pata e da alodinia ao frio, quando era aplicada acetona também na pata. No modelo inflamatório foi observada uma maior sensibilidade nos primeiros dias após a indução de dor, tendo os ratos apresentado uma recuperação parcial da sensibilidade no final do tempo de progressão do modelo. No modelo neuropático, os ratos modelo desenvolveram hiperalgesia durante o curso da experiência, durante o qual também desenvolveram alodinia ao frio. Ao fim de 21 e 28 dias após a indução de dor, nos modelos inflamatório e neuropático, respetivamente, os ratos foram sacrificados e os DRGs, foram coletados para futuras experiências. As propriedades biofísicas das correntes de Na+ foram estudadas por registos eletrofisiológicos em neurónios-DRG de pequeno diâmetro isolados, obtidos a partir de preparações ex-vivo de ratos controlo e dos modelos de DC. No estudo da DC inflamatória não se observaram diferenças significativas entre as correntes de Na+ provenientes de neurónios de ratos controlo e de dor. No entanto, no caso do estudo da dor neuropática, observaram-se algumas tendências: a densidade da corrente de Na+ que flui pela área membrana celular é maior nos neurónios-DRG provenientes dos ratos CCI e o tempo que essas mesmas correntes demoram a recuperar a sua magnitude após a inativação dos VGSC é também menor. Estas alterações biofísicas podem ser a razão pela qual, no contexto de DC, os neurónios encontram-se hiperexcitáveis e conseguem suportar uma maior frequência de disparo dos potenciais de ação. É importante referir que trabalhos anteriores demonstraram que o Nav1.8 é a isoforma presente em situações fisiológicas nos pequenos neurónios DRG, com um perfil eletrofisiológico que mais se adequa a estas alterações observadas. Paralelamente, a expressão proteica dos canais de sódio Nav1.7 e Nav1.8 foi comparada entre os gânglios do lado ipsilateral (sujeitos ao modelo de dor) e do lado contralateral (controlo interno) à indução de dor. Pela análise por Western blot, observou-se um aumento da expressão de Nav1.7 e Nav1.8 nos gânglios ipsilaterais dos ratos com dor neuropática (CCI). Por análise imunohistoquímica, observou-se que estes canais são expressos nos neurónios DRG e que poderá existir um aumento da expressão de Nav1.7 nos gânglios do lado ipsilateral no modelo inflamatório (CFA), quando comparados com os gânglios contralaterais. Em resumo, a presente dissertação permitiu observar que neurónios-DRG de pequeno diâmetro sofrem alterações na DC, nomeadamente nas propriedades biofísicas e de expressão dos VGSC. No futuro serão necessários mais ensaios eletrofisiológicos que discriminem dentro das correntes totais de Na+, quais as isoformas dos VGSC presentes nos neurónios DRG que induzem a modificações na sua excitabilidade. Deste modo, poder-se-á compreender quais os padrões-tipo de atividade bem como as diferenças a nível funcional dos neurónios provenientes de ratos com dor comparativamente aos controlos. Serão também necessárias mais análises de expressão dos VGSC nos DRGs destes modelos de dor, para um melhor entendimento das tendências observadas. Deste modo, espera-se que uma mais consistente validação dos neurónios DRG como alvo farmacológico e, em particular dos VGSC por estes expressos, tragam informações relevantes para o desenvolvimento de novas terapias para a DC. Idealmente, o fármaco permitiria aos pacientes a recuperação da excitabilidade fisiológica destes neurónios, sem atuar no sistema nervoso central, evitando efeitos secundários indesejáveis e conferido aos pacientes uma melhor qualidade de vida.
Chronic pain (CP) affects 21% of the human population and to date there is still no adequate treatment for this syndrome. Previous studies have indicated dorsal root ganglia (DRG) neurons, as crucial for the transmission of pain stimulus. The activity of these neurons leads to brain perception of pain, which is found to be exacerbated in CP patients. Such condition derives from the abnormal function of pivotal ion channels that shape the electric activity in DRG neurons. Therefore, the most promising therapeutic approach for CP relies on reducing this neuronal hyperexcitability. In this regard, voltage-gated Sodium channels (VGSC) have the ability to initiate the electrical signalling in excitable cells, a characteristic that makes them potential drug targets. The peripheral localization of DRG neurons and its preferential expression of the Sodium channel isoforms Nav1.7, Nav1.8 and Nav1.9, makes them key targets for the development of novel analgesics to treat CP. Nevertheless, there is still a great demand for novel functional studies of those currents/channels sub-types specifically involved in the processes of CP, namely neuropathic and inflammatory pain. The aim of this project was to study alterations in VGSC currents/channels in DRG neurons from rat models of neuropathic and inflammatory CP; specifically, to identify the biophysical and pharmacologic patterns that change following CP and correlating those with animal behaviour tests. Overall, results show development of hyperalgesia and an abnormal function and expression of VGSC in DRG neurons following neuropathic pain in rat models, contributing to the validation of VGSC in DRG neurons as potential targets for novel analgesics in CP.

Skeletal muscle weakness is a common problem that complicates recovery from critical illness. The primary causes of weakness include neuropathic disorders, myopathic disorders, and mixed disorders. Recent studies have demonstrated that reduced excitability of the nerve and muscle cell membranes might contribute to weakness during the acute stages of the polyneuropathy and myopathy encountered in critically ill patients. In both tissues, an acquired sodium channelopathy can lead to increased inactivation of channels, leading to inexcitability an paralysis. Experimental sepsis models have demonstrated a similar reduction in excitability in myocardial cells as well as in motor neurons within the spinal cord. The presence of a channelopathy in multiple tissues raises the possibility that reduced excitability of neurons within the CNS might contribute to septic encephalopathy. If this is the case, a single therapy to improve excitability might treat failure of a number of electrically active tissues.

Cerebral ischemia or hypoxia-ischemia initiate a cascade of biochemical events including impaired reuptake of glutamate into perisynaptic glia causing glutamate flooding, calcium fluxing through NMDA glutamate channels, activation of neuronal nitric oxide synthetase, and impaired mitochondrial ATP production. In animal models it is possible to block these steps and protect the brain but the temporal window of protection after the insult lasts only a few hours. Recombinant TPA is clinically protective if given within 3 hours of stroke, but other agents have not been shown to protect brain tissue after stroke. However, total body cooling has also been shown to protect the brain of term infants if initiated within 6 hours of perinatal asphyxia, and a similar level of cooling may provide protection for the brain in adults who have been resuscitated after cardiac arrest.

SOCRAT-BN is a software package to simulate design and severe accidents of sodium-cooled fast reactors. The package consists of modules for calculating damage to the reactor’s core, thermohydraulic processes and neutron physics. The thermohydraulic module has been developed to calculate one- and two-phase flows in channels with different geometry and bundles. The module is based on a two-fluid model for equal pressures of phases. In this paper we present an explanation of the deciding constitutive models for equations used in the system. Validation of the module was performed on the experimental data for one- and two-fluid flows in complex geometry channels and on calculation of running a first loop of the reactor BN-600 in nominal mode.

In this paper the thermal-hydraulic characteristics of the primary loop of the Experimental Breeder Reactor (EBR-II), including the temperature and the flow characteristics of the core, the intermediate heat exchanger (IHX) and the experiment subassembly XX09 and XX10, were analyzed with the transient thermal-hydraulic code THACS. The THACS code contains the core, the pumps, IHX, the sodium pool and some other modules, and each module could operate separately. All of the primary–loop components are simulated one-dimensional, and in the core calculation the incompressible model for the single phase. The multiple-channel model is applied to simulate the core subassemblies, including the average, hot, XX09, XX10, the reflector and the blanket channels. The neutron physics is calculated with the point reactor kinetics, and the reactivity feedbacks caused by the Doppler effect, coolant density, axial expansion of fuel rods and radial expansion of core are considered. Two tests, namely the SHRT-17 and SHRT-45R tests, are simulated to validate our tools and models. The THACS simulation results show that the EBR-II type sodium cooled fast reactor could shut down automatically relying on inherent negative feedbacks in the two tests.

Thermal striping on the core instrumentation plate (CIP) at the bottom of the upper internal structure (UIS) of an advanced loop-type sodium-cooled fast reactor in Japan (Advanced-SFR) has been numerically investigated. At the top of the core below the CIP, the sodium at high temperature flows out from the fuel subassemblies (FSs) and the sodium at low temperature flows out from the primary control rod (PCR) and backup control rod (BCR) channels, and also the radial blanket fuel subassemblies (RBFSs) at the outer side of the core. In order to predict the thermal striping on the CIP caused by mixing fluids at different temperatures from the FSs, the PCR and the BCR channels, and the RBFSs, a numerical estimation method using a spatial connection methodology between the upper plenum analysis and the local area analysis for the target area has been developed. By using the connection methodology, the numerical simulation considering the influence of the transversal flow in the UIS and the external flow around the UIS in the upper plenum can be performed to improve the accuracy of the estimation results. In this paper, the outline of the spatial connection methodology including data transfer technique from the upper plenum analysis to the local area analysis was described. As a validation process, numerical simulation of the water experiment using the test apparatus named TAFUT which was 1/3-scaled 1/6 partial model of the upper plenum of the Advanced-SFR was performed to confirm applicability of the spatial connection methodology to a practical thermal striping problem. The numerical result of temperature distribution was compared with the measured result in TAFUT experiment. Additionally, mesh sensitivity of the local area analysis model to the numerical results was indicated by using a small and a large area models in order to suggest an appropriate local area analysis model.

Electrokinetically driven flow of electrolyte solutions through micro- and nanochannels is of interest in microelectromechanical systems (MEMS) and nanotechnology applications. In this work, fully developed and steady electroosmotic flow (EOF) of dilute sodium tetraborate and sodium chloride aqueous solutions in a rectangular channel where the channel hight h is comparable to its width W is examined. EOF is also studied under conditions of electric double layer (EDL) overlap, or λ/h ∼ O(1), where λ is the Debye thickness, for very dilute solutions. The initial experimental data and model results are in very good agreement for dilute sodium tetraborate solutions. The experimental work uses the new nano-particle image velocimetry (nPIV) technique. Evanescent waves from the total internal reflection of light with a wavelength of 488 nm at a refractive index interface is used to illuminate 100 nm neutrally buoyant fluorescent particles in the near-wall region of the flow. The images of these tracer particles over time are processed to obtain the two components of the velocity field parallel to the wall in fully developed EOF of sodium tetraborate at concentrations up to 2 mM in fused quartz rectangular channels with height h up to 10 microns. The spatial resolution of these velocity field data along the dimension normal to the wall is about 100 nm, and the data are obtained within a distance of approximately 100 nm of the wall based upon the 1/e intensity point, or penetration depth. A set of equations modeling EOF in a long channel are solved where h/L << 1, and L is the lengthscale along the flow direction. Unlike most previous models, this work does not use the Debye-Huckel approximation, nor does it assume symmetric boundary conditions. For the case where λ/h << 1, analytical solutions for the velocity, potential and mole fractions are obtained using an asymptotic perturbation approach.

The blanket assemblies of China Experimental Fast Reactor (CEFR) were specially designed for the purpose of fuel breeding. There are two parts of the blanket assembly that have significant influence on its thermal-hydraulic feature: the labyrinth throttle set and the wire-wrapped fuel bundle. In this paper, the flow and temperature distribution of sodium coolant in fuel bundle, as well as the pressure drop in the labyrinth throttle set were investigated by solving the three-dimensional conservation equations of mass, momentum and energy, for a wide range of Reynolds number. Different turbulent models: k-ε, SST and Reynolds stress model (RSM) were used and compared with experimental correlations in the study of fuel bundle. The cross flow strength in three typical sub-channels and the influence of steer rods were also investigated. It has been found that the SST model fits best with the experimental correlation and although dampened the cross flow in edge channel, the steel rods installed in the hexagonal duct could enhance the heat transfer by “pushing” the sodium coolant to inner channel of the fuel bundle. Moreover, the relationship between the slot width of the labyrinth throttle set and the pressure drop under different coolant mass flow rate was analyzed, and found out the slot width that satisfied the design requirement of the blanket assembly.

Drug development platforms such as two-dimensional (2D) in vitro cell culture systems and in vivo animal studies do not accurately predict human in vivo effectiveness of candidate therapeutics [1]. Cell culture systems have limited similarities to primary human cells and tissues as only one cell type is employed and animal studies have a generally limited ability to recapitulate human drug response as different species have differences in metabolism, physiology, and behavior. Mike Leavitt, a former U.S. Secretary of Health and Human Services, has stated that “currently, nine out of ten experimental drugs fail in clinical studies because we cannot accurately predict how they will behave in people based on laboratory and animal studies” [2]. Therefore, this research project is focused on developing an in vitro platform to test candidate therapeutics for more efficacious predictions of human response. We have fabricated a three-dimensional (3D) breast cancer tissue volume containing a vascular network. This vascular network is necessary because current in vitro systems (e.g., rotating bioreactors, suspension of spheroids, and growth on a porous scaffold) are limited in size (1–2 mm) by their absence of micrometer-scale blood flow micro-channels that allow for oxygen and nutrient diffusion into the tissue [4]. The extracellular matrix scaffold has been developed to mimic the native extracellular matrix and includes relevant cell types (e.g., human breast cancer epithelial cells and human breast fibroblasts) along with the prefabricated vascular network (prevascularization). These systems are intended to support long-term growth, recapitulate physiological tissue function, and accurately model response to treatment. It is hypothesized that the development of reproducible tissue volumes will transform breast cancer drug development by providing reliable, cost-effective models that can more accurately predict therapeutic efficacy than current preclinical in vivo and in vitro models.

The ability to fabricate networks of micro-channels that obey the biological properties of bifurcating structures found in nature suggests that it is possible to construct artificial vasculatures or bronchial structures. These devices could aid in the desirable objective of eliminating many forms of animal testing. In addition, the ability to precisely control hydraulic conductance could allow designers to create specific concentration gradients that would allow biologists to correlate the behavior of cells. In 1926, Murray found that there was an optimum branching ratio between the diameters of the parent and daughter vessels at a bifurcation. For biological vascular systems, this is referred to as Murray’s law and its basic principle has been found to be valid in many plant and mammalian organisms. An important consequence arises from this law: when the successive generations consist of regular dichotomies, the tangential shear stress at the wall remains constant throughout the network. This simple concept forms an elegant biomimetic design rule that will allow designers to create complex sections with the desired hydraulic conductance or resistance. The paper presents a theoretical analysis of how biomimetic networks of constant-depth rectangular channels can be fabricated using standard photolithographic techniques. In addition, the design rule developed from Murray’s law is extended to a simple power-law fluid to investigate whether it is feasible to design biomimetic networks for non-Newtonian liquids. Remarkably, Murray’s law is obeyed for power-law fluids in cylindrical pipes. Although highly promising, the extension of the analysis to rectangular or trapezoidal channels requires much further work. Moreover, it is unclear at this stage whether Murray’s law holds for other non-Newtonian models.

A detailed safety assessment of innovative Generation IV reactor designs with heavy-liquid metal coolants, such as lead and lead-bismuth eutectic (LBE), requires an evaluation of the maximum core temperature in several postulated scenarios. Considering the complex geometry of fuel assemblies (FAs), and the low Prandtl number of the coolants, this flow scenario is challenging for the models used in numerical simulations, e.g. for relating the turbulent transport of momentum and heat. Thus, reliable experimental data are needed for validation. In recent years, a series of comprehensive heat transfer tests in fuel pin bundle simulators was performed at ENEA (Italy) and KIT (Germany) in the framework of the European collaborative projects THINS and SEARCH. Both grid and wire spacer geometries are considered, in a wide range of operating conditions (temperature, flow velocity and power density) representative of the ALFRED and MYRRHA fuel assemblies, in natural and forced-convective flow regimes. Although different experimental approaches were followed by each group (e.g. thermocouple position and average data treatment), there is a relatively good agreement on results in the overlapping regions. These experimental studies indicate that the mean Nusselt number is in well agreement with the predictions of empirical correlations developed for sodium systems. In particular, for wire-spaced FA, heat transfer results show values close to the Kazimi-Carelli correlation both for low and high flow rates at ENEA and KIT respectively. For grid-spaced FA, results are more in agreement with Ushakov correlation. Furthermore, large temperature differences are measured by thermo-couples installed at selected rods and sub-channels. A discussion on the influences of the spacer design and bundle size is included. This wide comparison allows an overview of the research on the HLM cooled fuel assembly in Europe.