Electrical stimulation of neonatal cardiomyocytes results in the sequential activation of nuclear genes governing mitochondrial proliferation and differentiation

Electrical stimulation of neonatal cardiac myocytes produces hypertrophy and cellular maturation with increased mitochondrial content and activity. To investigate the patterns of gene expression associated with these processes, cardiac myocytes were stimulated for varying times up to 72 hr in serum-free culture. The mRNA contents for genes associated with transcriptional activation [c-fos, c-jun, JunB, nuclear respiratory factor 1 (NRF-1)], mitochondrial proliferation [cytochrome c (Cyt c), cytochrome oxidase], and mitochondrial differentiation [carnitine palmitoyltransferase I (CPT-I) isoforms] were measured. The results establish a temporal pattern of mRNA induction beginning with c-fos (0.25–3 hr) and followed sequentially by c-jun (0.5–3 hr), JunB (0.5–6 hr), NRF-1 (1–12 hr), Cyt c (12–72 hr), and muscle-specific CPT-I (48–72 hr). Induction of the latter was accompanied by a marked decrease in the liver-specific CPT-I mRNA, thus supporting the developmental fidelity of this pattern of gene regulation. Consistent with a transcriptional mechanism, electrical stimulation increased c-fos, β-myosin heavy chain, and Cyt c promoter activities. These increases coincided with a rise in their respective endogenous gene transcripts. NRF-1, cAMP response element, and Sp-1 site mutations within the Cyt c promoter reduced luciferase expression in both stimulated and nonstimulated myocytes. Mutations in the NRF-1 and CRE sites inhibited the induction by electrical stimulation (5-fold and 2-fold, respectively) whereas mutation of the Sp-1 site maintained or increased the fold induction. This finding is consistent with the appearance of NRF-1 and fos/jun mRNAs prior to that of Cyt c and suggests that induction of these transcription factors is a prerequisite for the transcriptional activation of Cyt c expression. These results support a regulatory role for NRF-1 and possibly AP-1 in the initiation of mitochondrial proliferation.

Non-CpG methylation is prevalent in embryonic stem cells and may be mediated by DNA methyltransferase 3a

Current evidence indicates that methylation of cytosine in mammalian DNA is restricted to both strands of the symmetrical sequence CpG, although there have been sporadic reports that sequences other than CpG may also be methylated. We have used a dual-labeling nearest neighbor technique and bisulphite genomic sequencing methods to investigate the nearest neighbors of 5-methylcytosine residues in mammalian DNA. We find that embryonic stem cells, but not somatic tissues, have significant cytosine-5 methylation at CpA and, to a lesser extent, at CpT. As the expression of the de novo methyltransferase Dnmt3a correlates well with the presence of non-CpG methylation, we asked whether Dnmt3a might be responsible for this modification. Analysis of genomic methylation in transgenic Drosophila expressing Dnmt3a reveals that Dnmt3a is predominantly a CpG methylase but also is able to induce methylation at CpA and at CpT.

Fetal and maternal contributions to risk of pre-eclampsia: population based study

Objective: To use familial patterns of recurrence of pre-eclampsia to investigate whether paternal genes expressed in the fetus contribute to the mother’s risk of pre-eclampsia and whether mother’s susceptibility to pre-eclampsia is related to maternal inheritance by mitochondrial DNA.

Latent N-methyl-D-aspartate receptors in the recurrent excitatory pathway between hippocampal CA1 pyramidal neurons: Ca(2+)-dependent activation by blocking A1 adenosine receptors.

When performed at increased external [Ca2+]/[Mg2+] ratio (2.5 mM/0.5 mM), temporary block of A1 adenosine receptors in hippocampus [by 8-cyclopentyltheophylline (CPT)] leads to a dramatic and irreversible change in the excitatory postsynaptic current (EPSC) evoked by Schaffer collateral/commissural (SCC) stimulation and recorded by in situ patch clamp in CA1 pyramidal neurons. The duration of the EPSC becomes stimulus dependent, increasing with increase in stimulus strength. The later occurring component of the EPSC is carried through N-methyl-D-aspartate (NMDA) receptor-operated channels but disappears under either the NMDA antagonist 2-amino-5-phosphonovaleric acid (APV) or the non-NMDA antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX). These findings indicate that the late component of the SCC-evoked EPSC is polysynaptic: predominantly non-NMDA receptor-mediated SCC inputs excite CA1 neurons that recurrently excite each other by predominantly NDMA receptor-mediated synapses. These recurrent connections are normally silent but become active after CPT treatment, leading to enhancement of the late component of the EPSC. The activity of these connections is maintained for at least 2 hr after CPT removal. When all functional NMDA receptors are blocked by dizocilpine maleate (MK-801), subsequent application of CPT leads to a partial reappearance of NMDA receptor-mediated EPSCs evoked by SCC stimulation, indicating that latent NMDA receptors are recruited. Altogether, these findings indicate the existence of a powerful system of NMDA receptor-mediated synaptic contacts in SCC input to hippocampal CA1 pyramidal neurons and probably also in reciprocal connections between these neurons, which in the usual preparation are kept latent by activity of A1 receptors.

Superluminal sources.

Predictions for the apparent velocity statistics under simple beaming models are presented and compared to the observations. The potential applications for tests of unification models and for cosmology (source counts, measurements of the Hubble constant H0 and the deceleration parameter q0) are discussed. First results from a large homogeneous survey are presented. The data do not show compelling evidence for the existence of intrinsically different populations of galaxies, BL Lacertae objects, or quasars. Apparent velocities betaapp in the range 1-5 h-1, where h = H0/100 km.s-1.Mpc-1 [1 megaparsec (Mpc) = 3.09 x 10(22) m], occur with roughly equal frequency; higher values, up to betaapp = 10 h-1, are rather more scarce than appeared to be the case from earlier work, which evidently concentrated on sources that are not representative of the general population. The betaapp distribution suggests that there might be a skewed distribution of Lorentz factors over the sample, with a peak at gammab approximately 2 h-1 and a tail up to at least gammab approximately 10 h-1. There appears to be a clearly rising upper envelope to the betaapp distribution when plotted as a function of observed 5-GHz luminosity; a combination of source counts and the apparent velocity statistics in a larger sample could provide much insight into the properties of radio jet sources.

Interaction of an alkylating camptothecin derivative with a DNA base at topoisomerase I-DNA cleavage sites.

DNA topoisomerase I (top1) is a ubiquitous nuclear enzyme. It is specifically inhibited by camptothecin, a natural product derived from the bark of the tree Camptotheca acuminata. Camptothecin and several of its derivatives are presently in clinical trial and exhibit remarkable anticancer activity. The present study is a further investigation of the molecular interactions between the drug and the enzyme-DNA complex. We utilized an alkylating camptothecin derivative, 7-chloromethyl-10,11-methylenedioxycamptothecin (7-ClMe-MDO-CPT), and compared its activity against calf thymus top1 in a DNA oligonucleotide containing a single top1 cleavage site with the activity of its nonalkylating analog, 7-ethyl-10,11-methylenedioxycamptothecin (7-Et-MDO-CPT). In the presence of top1, 7-ClMe-MDO-CPT produced a DNA fragment that migrated more slowly than the top1-cleaved DNA fragment observed with 7-Et-MDO-CPT. Top1 was unable to religate this fragment in the presence of high NaCl concentration or proteinase K at 50 degrees C. This fragment was resistant to piperidine treatment and was also formed with an oligonucleotide containing a 7-deazaguanine at the 5' terminus of the top1-cleaved DNA (base + 1). It was however cleaved by formic acid treatment followed by piperidine. These observations are consistent with alkylation of the +1 base (adenine or guanine) by 7-ClMe-MDO-CPT in the presence of top1 covalent complexes and provide direct evidence that camptothecins inhibit top1 by binding at the enzyme-DNA interface.

Quantisation of the bosonic string

In this work we review the basic principles of the theory of the relativistic bosonic string through the study of the action functionals of Nambu-Goto and Polyakov and the techniques required for their canonical, light-cone, and path-integral quantisation. For this purpose, we briefly review the main properties of the gauge symmetries and conformal field theory involved in the techniques studied.

Adaptação do ensaio SPT para determinação de parâmetros adicionais para o dimensionamento de fundações

A presente pesquisa foi desenvolvida com o intuito de agregar alguns procedimentos ao Ensaio SPT, para se obter um maior número de parâmetros para o dimensionamento de fundações, visto que este ensaio é o tipo de investigação de subsolo mais utilizado no Brasil para este fim. Os procedimentos adicionados foram: medida de eficiência do equipamento e ensaio de arrancamento do amostrador em cada camada distinta de solo atravessado. Foram realizados nove ensaios de arrancamento do amostrador, em três furos de sondagem, nas profundidades de dois, cinco e dez metros (três camadas de solos distintas). Em um quarto furo foram realizadas duas provas de carga à compressão no amostrador (nas profundidades de dois e cinco metros) para determinação da eficiência do equipamento. Por meio destes ensaios foi possível obter: a resistência de ponta do ensaio SPT, a resistência por atrito lateral na face externa do amostrador, e a razão de atrito do ensaio SPT, que pode ser usada para a classificação de solo de modo similar ao utilizado para o ensaio de cone. Além destes resultados, neste trabalho foi também analisada a relação entre a resistência por atrito interna e externa no amostrador durante o ensaio, conhecida por a. Esta relação mostrou-se maior para a camada de solo com grande porcentagem de pedregulho e menor para a camada que apresentava maior porcentagem de argila. Foi observado também que o valor de a cresce linearmente com a razão entre o valor de NSPT e a tensão vertical efetiva na profundidade do ensaio. Os resultados deste trabalho também mostraram que o valor da razão entre a resistência de ponta do ensaio SPT (qSPT) e o valor de NSPT é único para o mesmo tipo de solo. Além disso, os valores encontrados nesta pesquisa da razão qSPT/ NSPT são próximos aos valores da razão entre a resistência de ponta do ensaio de cone e o NSPT encontrados na literatura.

Re-Examining the Stop-Signal Task to Test Competing Theories of AD/HD

The current study tested two competing models of Attention-Deficit/Hyperactivity Disorder (AD/HD), the inhibition and state regulation theories, by conducting fine-grained analyses of the Stop-Signal Task and another putative measure of behavioral inhibition, the Gordon Continuous Performance Test (G-CPT), in a large sample of children and adolescents. The inhibition theory posits that performance on these tasks reflects increased difficulties for AD/HD participants to inhibit prepotent responses. The model predicts that putative stop-signal reaction time (SSRT) group differences on the Stop-Signal Task will be primarily related to AD/HD participants requiring more warning than control participants to inhibit to the stop-signal and emphasizes the relative importance of commission errors, particularly "impulsive" type commissions, over other error types on the G-CPT. The state regulation theory, on the other hand, proposes response variability due to difficulties maintaining an optimal state of arousal as the primary deficit in AD/HD. This model predicts that SSRT differences will be more attributable to slower and/or more variable reaction time (RT) in the AD/HD group, as opposed to reflecting inhibitory deficits. State regulation assumptions also emphasize the relative importance of omission errors and "slow processing" type commissions over other error types on the G-CPT. Overall, results of Stop-Signal Task analyses were more supportive of state regulation predictions and showed that greater response variability (i.e., SDRT) in the AD/HD group was not reducible to slow mean reaction time (MRT) and that response variability made a larger contribution to increased SSRT in the AD/HD group than inhibitory processes. Examined further, ex-Gaussian analyses of Stop-Signal Task go-trial RT distributions revealed that increased variability in the AD/HD group was not due solely to a few excessively long RTs in the tail of the AD/HD distribution (i.e., tau), but rather indicated the importance of response variability throughout AD/HD group performance on the Stop-Signal Task, as well as the notable sensitivity of ex-Gaussian analyses to variability in data screening procedures. Results of G-CPT analyses indicated some support for the inhibition model, although error type analyses failed to further differentiate the theories. Finally, inclusion of primary variables of interest in exploratory factor analysis with other neurocognitive predictors of AD/HD indicated response variability as a separable construct and further supported its role in Stop-Signal Task performance. Response variability did not, however, make a unique contribution to the prediction of AD/HD symptoms beyond measures of motor processing speed in multiple deficit regression analyses. Results have implications for the interpretation of the processes reflected in widely-used variables in the AD/HD literature, as well as for the theoretical understanding of AD/HD.

Relationship between piezometric level and ground deformations measured by means of DInSAR in the Vega Media of the Segura River (Spain)

Differential SAR Interferometry (DInSAR) is a remote sensing method with the well demonstrated ability to monitor geological hazards like earthquakes, landslides and subsidence. Among all these hazards, subsidence involves the settlement of the ground surface affecting wide areas. Frequently, subsidence is induced by overexploitation of aquifers and constitutes a common problem that affects developed societies. The excessive pumping of underground water decreases the piezometric level in the subsoil and, as a consequence, increases the effective stresses with depth causing a consolidation of the soil column. This consolidation originates a settlement of ground surface that must be withstood by civil structures built on these areas. In this paper we make use of an advanced DInSAR approach - the Coherent Pixels Technique (CPT) [1] - to monitor subsidence induced by aquifer overexploitation in the Vega Media of the Segura River (SE Spain) from 1993 to the present. 28 ERS-1/2 scenes covering a time interval of about 10 years were used to study this phenomenon. The deformation map retrieved with CPT technique shows settlements of up to 80 mm at some points of the studied zone. These values agree with data obtained by means of borehole extensometers, but not with the distribution of damaged buildings, well points and basements, because the occurrence of damages also depends on the structural quality of the buildings and their foundations. The most interesting relationship observed is the one existing between piezometric changes, settlement evolution and local geology. Three main patterns of ground surface and piezometric level behaviour have been distinguished for the study zone during this period: 1) areas where deformation occurs while ground conditions remain altered (recent deformable sediments), 2) areas with no deformation (old and non-deformable materials), and 3) areas where ground deformation mimics piezometric level changes (expansive soils). The temporal relationship between deformation patterns and soil characteristics has been analysed in this work, showing a delay between them. Moreover, this technique has allowed the measurement of ground subsidence for a period (1993-1995) where no instrument information was available.

Monitorización de la subsidencia del terreno en la Vega Media del río Segura mediante Interferometría SAR diferencial (DInSAR)

Ground subsidence caused by aquifer exploitation is a geotechnical hazard that affects wide areas, causing important economic damages. This phenomenon is due to soil consolidation produced by the increase of effective stress caused by piezometric depletion. In this work a Remote Sensing Technique called Coherent Pixel (CPT) is applied to monitor subsidence in the Vega Media of the Segura River during a period of eleven years.

Istanbul, Turkey, 1882 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Plan von Constantinopel mit den Vorstädten, dem Hafen, und einem Theile des Bosporus = Plan de Constantinople avec ses faubourgs, le port et une partie du Bosphore, aufgenommen und gezeichnet von C. Stolpe nach den zeitherigen Veränderungen berichtigt bis zum Jahre 1882. It was published by Verlag von Lorentz & Keil in 1882. Scale 1:15,000. Covers Istanbul, Turkey. Map in German and French.The image inside the map neatline is georeferenced to the surface of the earth and fit to the 'European Datum 1950 UTM Zone 35N' coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map.This map shows features such as roads, railroads and stations, drainage, built-up areas (by ethnicity) and selected buildings, fortification, ground cover, parks, cemeteries, ruins, and more. Relief shown pictorially and by contours.This layer is part of a selection of digitally scanned and georeferenced historic maps from The Harvard Map Collection as part of the Imaging the Urban Environment project. Maps selected for this project represent major urban areas and cities of the world, at various time periods. These maps typically portray both natural and manmade features at a large scale. The selection represents a range of regions, originators, ground condition dates, scales, and purposes.

O papel simbólico da língua na construção das identidades nacionais : o caráter identitário da discussão sobre o AO90 em Portugal

Tese de doutoramento, Estudos de Literatura e de Cultura (Cultura e Comunicação), Universidade de Lisboa, Faculdade de Letras, 2016

Definizione del modello geotecnico e valutazione delle condizioni di stabilità di un tratto di argine del fiume Po, in località Viadana (MN).

Questa tesi si prefigge di analizzare le condizioni di stabilità di un argine inferiore del fiume Po situato presso il comune di Viadana (MN), più volte interessato da eventi franosi che si sono ripetuti con una certa frequenza nel corso degli ultimi anni. L'argomento proposto nella tesi prende spunto dall’esperienza presso il Polo Scientifico e Tecnologico di AIPO a Boretto (RE), nell’ambito del tirocinio formativo, durante il quale ho portato a termine un programma di prove geotecniche su campioni indisturbati prelevati nel sito interessato da fenomeni di instabilità, allo scopo di migliorare le conoscenze sulla risposta meccanica dei terreni arginali e del substrato di fondazione. Utilizzando i risultati delle prove di laboratorio, unitamente alle informazioni provenienti dai sondaggi e dalle prove penetrometriche (CPT/CPTU), è stato possibile innanzitutto identificare le unità stratigrafiche presenti nel sistema arginale e determinarne i parametri meccanici, per poi procedere alla costruzione del modello geotecnico di riferimento per le analisi di stabilità. Pertanto la tesi si pone come obiettivi principali la messa a punto di un modello geotecnico affidabile dell'argine, la verifica della stabilità globale delle scarpate lato fiume e l'individuazione dei fattori scatenanti i fenomeni di scorrimento osservati, nonché la scelta di possibili interventi per la messa in sicurezza delle sponde, valutandone i benefici in termini del fattore di sicurezza ed i relativi costi. L'insieme dei risultati conseguiti in questo studio permette di fornire utili indicazioni a coloro che sono preposti alla manutenzione e messa in sicurezza delle strutture arginali del fiume Po.