A nitric oxide synthase inhibitor decreases 6-hydroxydopamine effects on tyrosine hydroxylase and neuronal nitric oxide synthase in the rat nigrostriatal pathway

There is evidence that nitric oxide plays a role in the neurotransmitter balance within the basal ganglia and in the pathology of Parkinson`s disease. In the present work we investigated in striatal 6-hydroxydopamine (6-OHDA) lesioned rats the effects of a nitric oxide synthase (NOS) inhibitor, NG-nitro-L-arginine (L-NOARG), given systemically on both the dopaminergic (DA) neuronal loss and the neuronal NOS cell density. We analyzed the DA neuronal loss through tyrosine hydroxylase immunohistochemistry (TH). The nitrergic system was evaluated using an antibody against the neuronal NOS (nNOS) isoform. Treatment with the L-NOARG significantly reduced 6-OHDA-induced dopaminergic damage in the dorsal striatum, ventral substantia nigra and lateral globus pallidus, but had no effects in the dorsal substantia nigra and in the cingulate cortex. Furthermore, L-NOARG reduced 6-OHDA-induced striatal increase, and substantia nigra compacta decrease, in the density of neuronal nitric oxide synthase positive cells. These results suggest that nitric oxide synthase inhibition may decrease the toxic effects of 6-OHDA on dopaminergic terminals and on dopamine cell bodies in sub-regions of the SN and on neuronal nitric oxide synthase cell density in the rat brain. (c) 2008 Elsevier B.V. All rights reserved.

Differential perturbation of neuronal and glial glutamate transport systems in retinal ischaemia

Glutamate is the major excitatory neurotransmitter in the retina and is removed from the extracellular space by an energy-dependent process involving neuronal and glial cell transporters. The radial glial Muller cells express the glutamate transporter, GLAST, and preferentially accumulate glutamate. However, during an ischaemic episode, extracellular glutamate concentrations may rise to excitotoxic levels. Is this catastrophic rise in extracellular glutamate due to a failure of GLAST? Using immunocytochemistry, we monitored the transport of the glutamate transporter substrate, D-aspartate, in the retina under normal and ischaemic conditions. Two models of compromised retinal perfusion were compared: (1) Anaesthetised rats had their carotid arteries occluded for 7 days to produce a chronic reduction in retinal blood flow. Retinal function was assessed by electroretinography. D-aspartate was injected into the eye for 45 min, Following euthanasia, the retina was processed for D-aspartate. GLAST and glutamate immunocytochemistry. Although reduced retinal perfusion suppresses the electroretinogram b-wave, neither retinal histology, GLAST expression, nor the ability of Muller cells to uptake D-aspartate is affected. As this insult does not appear to cause excitotoxic neuronal damage, these data suggest that GLAST function and glutamate clearance are maintained during periods of reduced retinal perfusion. (2) Occlusion of the central retinal artery for 60 min abolishes retinal perfusion, inducing histological damage and electroretinogram suppression. Although GLAST expression appears to be normal. its ability to transport D-aspartate into Muller cells is greatly reduced. Interestingly, D-aspartate is transported into neuronal cells, i.e. photoreceptors, bipolar and ganglion cells. This suggests that while GLAST is vitally important for the clearance of excess extracellular glutamate, its capability to sustain inward transport is particularly susceptible to an acute ischaemic attack. Manipulation of GLAST function could alleviate the degeneration and blindness that result from ischaemic retinal disease. (C) 2001 Elsevier Science Ltd, All rights reserved.

A zebrafish homologue of deleted in colorectal cancer (zdcc) is expressed in the first neuronal clusters of the developing brain

DCC (deleted in colon cancer), Neogenin and UNC-5 are all members of the immunoglobulin superfamily of transmembrane receptors which are believed to play a role in axon guidance by binding to their ligands, the Netrin/UNC-40 family of secreted molecules (Cell. Mol. Life Sci. 56 (1999) 62; Curr. Opin. Genet. Dev. 7 (1997) 87). Although zebrafish homologues of the Netrin family of secreted molecules have been reported, to date there has been no published description of zebrafish DCC homologues (Mol. Cell. Neurosci. 9 (1997) 293., Mol. Cell. Neurosci. I I ( 1998) 194; Mech. Dev. 62 (1997) 147). We report here the expression pattern of a zebrafish dcc (zdcc) homologue during the initial period of neurogenesis and axon tract formation within the developing central nervous system. Between 12 and 33 h post-fertilisation zdcc is expressed in a dynamic spatiotemporal pattern in all major subdivisions of the central nervous system. Double-labelling for zdcc and the post-mitotic neuronal marker HNK-1 revealed that subpopulations of neurons within the first nuclei of the zebrafish brain express zdcc. These results support our previous observation that patterning of neuronal clusters in the zebrafish brain occurs early in development (Dev. Bioi, 229 (2001) 271). (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.

Suppressor of cytokine signaling 2 regulates neuronal differentiation by inhibiting growth hormone signaling

The intracellular mechanisms that determine the response of neural progenitor cells to growth factors and regulate their differentiation into either neurons or astrocytes remain unclear. We found that expression of SOCS2, an intracellular regulator of cytokine signaling, was restricted to mouse progenitor cells and neurons in response to leukemia inhibitory factor (LIF)-like cytokines. Progenitors lacking SOCS2 produced fewer neurons and more astrocytes in vitro, and Socs2(-/-) mice had fewer neurons and neurogenin-1 (Ngn1)-expressing cells in the developing cortex, whereas overexpression of SOCS2 increased neuronal differentiation. We also report that growth hormone inhibited Ngn1 expression and neuronal production, and this action was blocked by SOCS2 overexpression. These findings indicate that SOCS2 promotes neuronal differentiation by blocking growth hormone-mediated downregulation of Ngn1.

A molecular mechanism for mRNA trafficking in neuronal dendrites

Specific neuronal mRNAs are localized in dendrites, often concentrated in dendritic spines and spine synapses, where they are translated. The molecular mechanism of localization is mostly unknown. Here we have explored the roles of A2 response element (A2RE), a cis-acting signal for oligodendrocyte RNA trafficking, and its cognate trans-acting factor, heterogeneous nuclear ribonucleoprotein ( hnRNP) A2, in neurons. Fluorescently labeled chimeric RNAs containing A2RE were microinjected into hippocampal neurons, and RNA transport followed using confocal laser scanning microscopy. These RNA molecules, but not RNA lacking the A2RE sequence, were transported in granules to the distal neurites. hnRNP A2 protein was implicated as the cognate trans-acting factor: it was colocalized with RNA in cytoplasmic granules, and RNA trafficking in neurites was compromised by A2RE mutations that abrogate hnRNP A2 binding. Coinjection of antibodies to hnRNP A2 halved the number of trafficking cells, and treatment of neurons with antisense oligonucleotides also disrupted A2RE - RNA transport. Colchicine inhibited trafficking, whereas cells treated with cytochalasin were unaffected, implicating involvement of microtubules rather than microfilaments. A2RE-like sequences are found in a subset of dendritically localized mRNAs, which, together with these results, suggests that a molecular mechanism based on this cis-acting sequence may contribute to dendritic RNA localization.

Economic viability of the third milking in systems of production using closed-circuit mechanical milking

This study aimed to analyze the economic viability of the third milking in production systems using mechanical milking in a closed circuit, aiming to provide technicians and farmers with information to assist them in decision-making. Specifically, it intended: (a) to estimate the cost of one milking; (b) to estimate the cost of the third milking; (c) to develop a mathematical equation to estimate the minimum amount of milk produced with two milkings, from which it would be economically feasible to do the third milking. Data were collected from three dairy farms, from November 2010 to March 2011, keeping a twice-a-day milking frequency, with three data collections in each farm, totalizing nine collections. Considering the average data, it would be feasible to do the third milking if the average milk yield per day of lactating cows in a twice-a-day milking frequency was greater than or equal to 24.43 kg of milk.

A DC Voltage-Multiplier Circuit Working as a High-Voltage Pulse Generator

The intensive use of semiconductor devices enabled the development of a repetitive high-voltage pulse-generator topology from the dc voltage-multiplier (VM) concept. The proposed circuit is based on an odd VM-type circuit, where a number of dc capacitors share a common connection with different voltage ratings in each one, and the output voltage comes from a single capacitor. Standard VM rectifier and coupling diodes are used for charging the energy-storing capacitors, from an ac power supply, and two additional on/off semiconductors in each stage, to switch from the typical charging VM mode to a pulse mode with the dc energy-storing capacitors connected in series with the load. Results from a 2-kV experimental prototype with three stages, delivering a 10-mu s pulse with a 5-kHz repetition rate into a resistive load, are discussed. Additionally, the proposed circuit is compared against the solid-state Marx generator topology for the same peak input and output voltages.

A influência da hipertonia biomecânica VS neuronal na selecção das estratégias de intervenção em sujeitos com sequelas de acidente vascular encefálico no território da artéria cerebral média

Introdução: Após lesão do SNC o músculo pode perder a sua variabilidade e flexibilidade, tal como se verifica em indivíduos após AVE. A caracterização do tónus muscular tem sido um indicador a ter em atenção para o diagnóstico clínico. As alterações do tónus podem resultar de uma combinação de alterações neurais, como consequência dos processos inerentes à neuroplasticidade e alterações biomecânicas. Objectivo: Verificar quais as modificações no tónus muscular, segundo a escala de Tardieu, após a aplicação de um programa de reabilitação neuromotora baseado no conceito de Bobath em dois indivíduos com sequelas de AVE. Pretendeu-se também verificar as repercussões nas actividades funcionais. Participantes e métodos: Foram seleccionados dois indivíduos e aplicado um programa de reabilitação, durante onze semanas, e avaliados em dois momentos, antes da intervenção (PRE) e após a intervenção (APÓS). Aplicaram-se vários instrumentos de avaliação, nomeadamente a escala de Tardieu. O programa de reabilitação realizado baseou-se no conceito de Bobath. Resultados: Na escala de Tardieu, foi comum aos dois indivíduos melhorias a nível da qualidade de reacção muscular. Ambos os indivíduos apresentaram melhorias no controlo postural e equilíbrio, que se evidenciaram na CIF. Conclusão: Foi possível observar modificações no tónus muscular após aplicação de um programa de reabilitação e, consequentemente modificações na distribuição da carga na base de suporte, no alinhamento das estruturas articulares e musculares e na marcha. Ao longo da intervenção, observaram-se repercussões positivas em ambos os indivíduos, permitindo a estes realizar as AVDs com menor dificuldade

Photonics Active Filters Based on SiC Multi layer Structures: A Two Stage Active Circuit

Characteristics of tunable wavelength filters based on a-SiC:H multi-layered stacked cells are studied both theoretically and experimentally. Results show that the light-activated photonic device combines the demultiplexing operation with the simultaneous photodetection and self amplification of an optical signal. The sensor is a bias wavelength current-controlled device that make use of changes in the wavelength of the background to control the power delivered to the load, acting a photonic active filter. Its gain depends on the background wavelength that controls the electrical field profile across the device.

A DC-DC Step-Up mu-Power Converter for Energy Harvesting Applications, Using Maximum Power PointTracking, Based on Fractional Open Circuit Voltage

A DC-DC step-up micro power converter for solar energy harvesting applications is presented. The circuit is based on a switched-capacitorvoltage tripler architecture with MOSFET capacitors, which results in an, area approximately eight times smaller than using MiM capacitors for the 0.131mu m CMOS technology. In order to compensate for the loss of efficiency, due to the larger parasitic capacitances, a charge reutilization scheme is employed. The circuit is self-clocked, using a phase controller designed specifically to work with an amorphous silicon solar cell, in order to obtain themaximum available power from the cell. This will be done by tracking its maximum power point (MPPT) using the fractional open circuit voltage method. Electrical simulations of the circuit, together with an equivalent electrical model of an amorphous silicon solar cell, show that the circuit can deliver apower of 1132 mu W to the load, corresponding to a maximum efficiency of 66.81%.

A Voltage Limiter Circuit for Indoor Light Energy Harvesting Applications

A voltage limiter circuit for indoor light energy harvesting applications is presented. This circuit is a part of a bigger system, whose function is to harvest indoor light energy, process it and store it, so that it can be used at a later time. This processing consists on maximum power point tracking (MPPT) and stepping-up, of the voltage from the photovoltaic (PV) harvester cell. The circuit here described, ensures that even under strong illumination, the generated voltage will not exceed the limit allowed by the technology, avoiding the degradation, or destruction, of the integrated die. A prototype of the limiter circuit was designed in a 130 nm CMOS technology. The layout of the circuit has a total area of 23414 mu m(2). Simulation results, using Spectre, are presented.

Start-up circuit for low-power indoor light energy harvesting applications

A start-up circuit, used in a micro-power indoor light energy harvesting system, is described. This start-up circuit achieves two goals: first, to produce a reset signal, power-on-reset (POR), for the energy harvesting system, and secondly, to temporarily shunt the output of the photovoltaic (PV) cells, to the output node of the system, which is connected to a capacitor. This capacitor is charged to a suitable value, so that a voltage step-up converter starts operating, thus increasing the output voltage to a larger value than the one provided by the PV cells. A prototype of the circuit was manufactured in a 130 nm CMOS technology, occupying an area of only 0.019 mm(2). Experimental results demonstrate the correct operation of the circuit, being able to correctly start-up the system, even when having an input as low as 390 mV using, in this case, an estimated energy of only 5.3 pJ to produce the start-up.

Identification of neuronal alterations induced by Shank3 mutations using iPS cells from Phelan-McDermid Patients' Fibroblasts

A família de proteínas Shank é o principal conjunto de proteinas de suporte e está localizada na densidade pós-sináptica das sinapses excitatórias. Existem 3 genes na família Shank, Shank1, Shank2 e Shank3 e são caracterizados por múltiplos domínios repetidos de anquirina próximo ao N-terminal seguido pelos domínios Src homologo 3 e PDZ, uma região longa rica em prolina e um domínio de motivo α estéril próximo ao C-terminal. Shank proteínas conectam duas subunidades de receptors glutamatérgicos, recetores NMDA e recetores metabotrópicos de glutamato do tipo-I (mGluRs). O domínio PDZ da Shank conecta-se ao C-terminal do GKAP e este, liga-se, ao complexo recetor PSD-95-NMDA. Por outro lado, a proteína Homer interage com o domínio rico em prolina para confirmar a associação entre a proteína Shank com o mGluR tipo-I. A proteína específica em estudo, Shank3, é haploinsuficiente em pacientes com sindrome Phelan-McDermid devido à deleções no braço comprido do cromossoma 22 levando à danos intelectuais, ausência ou atraso no discurso, comportamentos semelhantes ao autismo, hipotonia e características dismórficas. Neste trabalho, investigamos o papel da Shank3 na função sináptica para compreender a relação entre alterações nesta proteína e as características neurológicas presente em Pacientes com síndrome Phelan-McDermid. Foram utilizados dois modelos diferentes, ratinhos knockout Shank3 e hiPSC de pacientes com PMS. Ratinhos geneticamente modificados são ferramentas uteis no estudo de genes e na compreensão dos mecanismos que experiências in vitro não são capazes de reproduzir, mas de maneira a compreender melhor as patologias humanas, decidimos trabalhar também com células humanas. Os fibroblastos dos pacientes com síndrome Phelan-McDermid fora reprogramados em hiPS cells, diferenciados em neurónios e comparados com os neurónios obtidos a partir de doadores saudavéis e da mesma idade. A reprogramação em iPSC foi realizada por infecção de lentivirus com quatro genes de reprogramação OCT4, c-MYC, SOX2 e KFL4 para posteriormente serem diferenciados em neurónios, com cada passo sendo positivamente confirmado através de marcadores neuronais. Através dos neurónios diferenciados, analisamos a expressão de proteínas sinápticas. Pacientes com haploinsuficiencia na proteína Shank3 apresentam níveis elevados de proteína mGluR5 e decrescidos de proteína Homer sugerindo que a haploinsuficiencia leva a desregulação do complexo mGluR5-Homer-Shank3 conduzindo também, a defeitos na maturação sináptica. Assim, a expressão da proteína mGluR5 está alterada nos pacientes com PMS podendo estar relacionada com defeitos encontrados na diferenciação neuronal e maturação sináptica observados nos neurónios de pacientes. Conclusivamente, iPS cells representam um modelo fundamental no estudo da proteína Shank3 e a sua influência no sindrome de Phelan-McDermid.