Dual ifgMosaic: A Versatile Method for Multispectral and Combinatorial Mosaic Gene-Function Analysis.

Improved methods for manipulating and analyzing gene function have provided a better understanding of how genes work during organ development and disease. Inducible functional genetic mosaics can be extraordinarily useful in the study of biological systems; however, this experimental approach is still rarely used in vertebrates. This is mainly due to technical difficulties in the assembly of large DNA constructs carrying multiple genes and regulatory elements and their targeting to the genome. In addition, mosaic phenotypic analysis, unlike classical single gene-function analysis, requires clear labeling and detection of multiple cell clones in the same tissue. Here, we describe several methods for the rapid generation of transgenic or gene-targeted mice and embryonic stem (ES) cell lines containing all the necessary elements for inducible, fluorescent, and functional genetic mosaic (ifgMosaic) analysis. This technology enables the interrogation of multiple and combinatorial gene function with high temporal and cellular resolution.

A high-fat diet combined with food deprivation increases food seeking and the expression of candidate biomarkers of addiction.

A mouse model has been developed to study the effect of dietary fat combined with food deprivation periods on palatable food seeking and on the expression of three potential addiction biomarkers in the nucleus accumbens: fumarate hydratase (FH), ATP synthase subunit alpha (ATP5a1) and transketolase (TKT). Forty C57BL/6 J male mice, four-week old, were fed either with a high-fat (HF) diet or standard diet along the experiment. After 3 weeks of differential feeding, animals underwent a two-week training period of two daily sessions where visual cues were paired either to palatable food (chocolate cereals) or no food at all. This training was prolonged one more week with similar, one daily sessions preceded by 12 hours of food deprivation. A behavioural test was finally conducted where mice were confined for 30 minutes either in food unpaired compartments or in compartments previously paired with cereals, but now with empty food trays. Total activity during this behavioural test and serum corticosterone levels right after it were similar in all experimental groups. Mice tested in food-paired compartments showed a marked preference for the empty food tray that gradually disappeared in standard diet-fed individuals but persisted in HF-fed mice. HF-fed mice also overexpressed FH, ATP5a1 and TKT, which positively correlated with the persistence of preference for the empty food tray. It is suggested that HF diets combined with food deprivation may enhance food seeking behaviours while upregulating FH/ATP5a1/TKT, which are further envisaged as biomarkers of addiction.

Towards a future molecular diagnosis of autism: Recent advances in biomarkers research from saliva samples.

Autism spectrum disorder diagnosis is currently based on clinical observations and behavioral evaluations exclusively, without any biological determination. Molecular biomarkers are usually obtained from biological fluids, such as blood or urine, generally through invasive and uncomfortable procedures. Patients with autism are characterized by sensory reactivity and behavioral difficulties which make sample collection problematic. Saliva has emerged as a feasible alternative to obtain relevant biological information and is especially indicated in the case of children with autism due to its painless and noninvasive sampling characteristics. Furthermore, saliva represents a valuable resource to study candidate biomarkers of autism. This has resulted in a number of interesting studies reported during the last 5 years that we have gathered and briefly discussed.

Proanthocyanidins block aldosterone-dependent up-regulation of cardiac gamma ENaC and Nedd4-2 inactivation via SGK1.

Aldosterone plays a central role in the development of cardiac pathological states involving ion transport imbalances, especially sodium transport. We have previously demonstrated a cardioprotective effect of proanthocyanidins in aldosterone-treated rats. Our objective was to investigate for the first time the effect of proanthocyanidins on serum and glucocorticoid-regulated kinase 1 (SGK1), epithelial Na+ channel (γ-ENaC), neuronal precursor cells expressed developmentally down-regulated 4-2 (Nedd4-2) and phosphoNedd4-2 protein expression in the hearts of aldosterone-treated rats. Male Wistar rats received aldosterone (1mg kg-1day-1)+1% NaCl for 3weeks. Half of the animals in each group were simultaneously treated with the proanthocyanidins-rich extract (80% w/w) (PRO80, 5mg kg-1day-1). Hypertension and diastolic dysfunction induced by aldosterone were abolished by treatment with PRO80. Expression of fibrotic, inflammatory and oxidative mediators were increased by aldosterone-salt administration and blunted by PRO80. Antioxidant capacity was improved by PRO80. The up-regulated aldosterone mediator SGK1, ENaC and p-Nedd4-2/total Nedd4-2 ratio were blocked by PRO80. PRO80 blunted aldosterone-mineralocorticoid-mediated up-regulation of ENaC provides new mechanistic insight of the beneficial effect of proanthocyanidins preventing the cardiac alterations induced by aldosterone excess.

Electroencefalografía cuantitativa como herramienta para el diagnóstico y seguimiento del paciente con trastorno por déficit de atención/hiperactividad / Quantitative electroencephalography as a tool for the diagnosis and follow-up of patients with attention-deficit/hyperactivity disorder

INTRODUCCIÓN: El diagnóstico del trastorno por déficit de atención/hiperactividad (TDAH) se basa en la presencia de síntomas conductuales referidos por el paciente o por sus cuidadores, lo que lleva asociado un alto componente subjetivo. Esto señala la necesidad de incorporar biomarcadores que ayuden a identificar objetivamente esta condición y contribuyan a valorar cuantitativamente la evolución del paciente y la eficacia de los tratamientos. Estudios publicados en los últimos años sugieren que los datos obtenidos mediante electroencefalografía cuantitativa (qEEG) podrían ser útiles para el diagnóstico y seguimiento del paciente con TDAH; sin embargo, muchos profesionales desconocen su potencial. OBJETIVOS: Describir y discutir los principales hallazgos de la neurometría mediante qEEG en sujetos con TDAH en los últimos 10 años. PACIENTES Y MÉTODOS: Se realizó una búsqueda sistemática basada en la metodología PRISMA sobre artículos originales publicados en los últimos 10 años e indexados en la base de datos PubMed, que aportaran datos obtenidos con qEEG en el contexto del TDAH. RESULTADOS: Se seleccionaron un total de 13 artículos de investigación de un total de 65 identificados inicialmente. CONCLUSIONES: Los parámetros de qEEG podrían aportar información relevante en el contexto del TDAH; sin embargo, su utilidad se limita actualmente a complementar otros métodos de diagnóstico subjetivo y no a su reemplazo, ya que los datos publicados son preliminares y se necesitan estudios más amplios y profundos. Sin embargo, el qEEG puede ayudar a entender mejor el TDAH, su neurofisiología, caracterizar sus subtipos y avanzar, por tanto, hacia tratamientos e intervenciones lo más personalizadas y efectivas posibles

INTRODUCTION: Attention-deficit/hyperactivity disorder (ADHD) diagnosis is based on behavioral symptoms referred by the patient or their caregivers, which includes a high subjective component. This highlights the need to find biomarkers that might help clinicians to objectively identify this condition and to assess the evolution of the patient and the effectiveness of the treatments using quantitative information. Previous studies suggest that data from quantitative electroencephalography (qEEG) could be useful for the diagnosis and follow-up of patients with ADHD; however, many professionals do not know its possibilities. AIMS. To describe and discuss the principal findings of qEEG neurometry in ADHD subjects along the last 10 years. PATIENTS AND METHODS: A systematic review, based on PRISMA methodology and using PubMed database, was carried out to identify original articles published in the last 10 years, that studied qEEG parameters within the ADHD context. RESULTS: A total of 13 research articles out of 65 were selected for an in-depth analysis. CONCLUSIONS: qEEG parameters could provide relevant information in the context of ADHD; however, its applicability is currently limited to complementing the subjective diagnosis methods and not to its replacement, as published data are just preliminary and need larger and deeper studies. However, qEEG data can help to better understand ADHD and its neurophysiology, characterize its subtypes and therefore move towards personalized and more effective treatments