Expression of the VEGF gene family during retinal vaso-obliteration and hypoxia.

Vascular insufficiency and retinal ischaemia precede many proliferative retinopathies and stimulate secretion of vasoactive growth factors. Vascular endothelial growth factor (VEGF) plays a major role and we therefore investigated the other members of the VEGF family: Placental growth factor (PlGF), VEGF-B, -C, and -D, and platelet derived growth factors (PDGF) A and B. Neonatal mice were exposed to hyperoxia for 5 days and then returned to room air (resulting in acute retinal ischaemia). RT-PCR demonstrated that all the members of the VEGF family are expressed in the retina and in situ hybridization (ISH) located their mRNAs primarily in ganglion cells. Similarly to VEGF itself, VEGF-C, PDGF-A, and PDGF-B were upregulated during retinal ischaemia (P < 0.05). Only PlGF gene expression increased during hyperoxia (P < 0.01). The expression pattern of these growth factors suggests a role in the normal retina and during vaso-obliterative and ischaemic phases.

eNOS overexpression exacerbates vascular closure in the obliterative phase of OIR and increases angiogenic drive in the subsequent proliferative stage

Purpose: In ischemic retinopathies, the misdirection of reparative angiogenesis away from the hypoxic retina leads to pathologic neovascularization. Thus, therapeutic strategies that reverse this trend would be extremely beneficial. Nitric oxide (NO) produced by endothelial nitric oxide synthase (eNOS) is an important mediator of vascular endothelial growth factor (VEGF) function facilitating vascular growth and maturation. However, in addition to NO, eNOS can also produce superoxide (O), exacerbating pathology. Here, our aim was to investigate the effect of eNOS overexpression on vascular closure and subsequent recovery of the ischemic retina.

Methods: Mice overexpressing eNOS-GFP were subjected to oxygen-induced retinopathy (OIR) and changes in retinal vascularization quantified. Background angiogenic drive was assessed during vascular development and in aortic rings. NOS activity was measured by Griess assay or conversion of radiolabeled arginine to citrulline, nitrotyrosine (NT), and superoxide by immunolabeling and dihydroethidium fluorescence and VEGF by ELISA.

Results: In response to hyperoxia, enhanced eNOS expression led to increased NOS-derived superoxide and dysfunctional NO production, NT accumulation, and exacerbated vessel closure associated with tetrahydrobiopterin (BH) insufficiency. Despite worse vaso-obliteration, eNOS overexpression resulted in elevated hypoxia-induced angiogenic drive, independent of VEGF production. This correlated with increased vascular branching similar to that observed in isolated aortas and during development. Enhanced recovery was also associated with neovascular tuft formation, which showed defective NO production and increased eNOS-derived superoxide and NT levels.

Conclusions: In hyperoxia, reduced BH bioavailability causes overexpressed eNOS to become dysfunctional, exacerbating vaso-obliteration. In the proliferative phase, however, eNOS has important prorepair functions enhancing angiogenic growth potential and recovery in ischemia. © 2012 The Association for Research in Vision and Ophthalmology, Inc.

Strategies for revascularizing the ischemic retina

Les rétinopathies ischémiques (RI) sont la cause majeure de cécité chez les personnes âgées de moins de 65 ans. Il existe deux types de RIs soit la rétinopathie du prématuré (ROP) ainsi que la rétinopathie diabétique (RD). Les RIs sont décrites en deux phases soit la phase de vasooblitération, marquée par une perte importante de vaisseaux sanguins, et une phase de néovascularisation secondaire à lʼischémie menant à une croissance pathologique de vaisseaux. Cette seconde phase peut générer des complications cliniques telles quʼun oedème dans lʼhumeur vitré ainsi que le détachement de la rétine chez les patients déjà atteints dʼune RI. Les traitements approuvés pour les RIs visent à réduire la formation des vaisseaux pathologiques ou lʼoedème; mais ceux-ci malheureusement ne règlent pas les problèmes sous-jacents tels que la perte vasculaire et lʼischémie. La rétine est un tissu hautement vascularisé qui contribue à lʼirrigation et à lʼhoméostasie des neurones. Lʼinteraction neurovasculaire, comprenant de neurones, vaisseaux et cellules gliales, contribue au maintien de cette homéostasie. Durant le développement, les neurones et les cellules gliales jouent un rôle important dans la vascularisation de la rétine en sécrétant des facteurs qui stimulent l'angiogenèse. Cependant, nos connaissances sur lʼinteraction neurovasculaire dans les RIs sont limitées. En identifiant les interactions importantes entre les cellules composant cette unité neurovasculaire dans la rétine, nous pourrons viser des cibles qui engendreront une revascularisation seine afin de diminuer les signes pathologiques chez les patients atteints dʼune RI. Les travaux présentés dans cette thèse visent à mieux expliquer cette interaction neurovasculaire en soulignant des concepts importants propres aux RIs. En utilisant un modèle de rétinopathie induite par lʼoxygène chez la souris, qui reproduit les caractéristiques importantes de la ROP (et en certaines instances, la RD), nous identifions quelques molécules clés jouant un rôle significatif dans les RIs soit la sémaphorine 3A (sema3A), lʼIL-1β, ainsi que le récepteur PAR2. Nos résultats démontrent que Sema3A, sécrétée par les cellules ganglionnaires rétiniennes (CGRs) durant une ischémie, empêche la revascularisation normale et que cette expression est induite par lʼIL-1β provenant des microglies activées. En bloquant Sema3A directement ou via lʼinhibition de lʼIL- 1β, nous remarquons une revascularisation seine ainsi quʼune diminution importante des vaisseaux pathologiques. Cela nous indique que Sema3A est impliquée dans la guidance vasculaire et quʼelle contribue à la pathogenèse des RIs. Lʼactivation de façon exogène de PAR2, identifié aussi comme régulateur du récepteur de lʼIL-1β (IL- 1RI) sur les CGRs, se traduit par une diminution séquentielle de lʼIL-1RI et de Sema3A ce qui mène également à une revascularisation seine. En conclusion, ces travaux soulignent lʼimportance de lʼinteraction neurovasculaire ainsi que la guidance vasculaire dans les RIs. Ils renforcent lʼimportance de la communication entre neurone, vaisseau et microglie dans la pathogenèse des RIs. Finalement, nous identifions quelques molécules clés qui pourront servir comme cibles afin de lutter contre lʼischémie qui cause des problèmes vasculaires chez les patients atteints dʼune RI.

Manualidades escolares televisivas : el vaso de papel.

Se dan indicaciones para la construcción de un vaso hecho con papel para ser utilizado en los recreos y excursiones. Es una lección ampliada del programa dedicado a manualidades emitido ya en la televisión escolar.

Vaso de planta

Este item é parte de um conjunto de modelos produzido pela equipe do Laboratório de Objetos de Apredizagem (LOA)/UFSCar para o jogo LabTecA

Condutividade elétrica da solução nutritiva para o cultivo do crisântemo em vaso

Em se tratando do crisântemo de vaso, que constitui relevante contribuição à atividade de plantas ornamentais, ainda há necessidade de uma recomendação consistente sobre a condutividade elétrica (CE) da solução nutritiva para seu cultivo. Nesse sentido, realizou-se um experimento em ambiente protegido, objetivando avaliar o crescimento do crisântemo cv. Miramar cultivado em vaso, em função da CE da solução nutritiva e da lixiviação de sais do substrato. O delineamento experimental foi em blocos casualizados, com quatro repetições; os tratamentos foram distribuídos em esquema fatorial 5 x 2, referentes aos valores de CE da solução nutritiva (2,1; 2,8; 3,5; 4,2; e 4,9 dS m-1), em substrato com e sem lavagem para lixiviação dos sais. O aumento da CE inibiu o crescimento e desempenho vegetativo, mas estimulou o diâmetro da haste. A lixiviação dos sais estimulou, exceto quanto ao diâmetro da haste, o maior crescimento avaliado pelas demais variáveis fitotécnicas. A CE de 2,1 dS m-1 possibilita a produção de crisântemo dentro de padrões qualitativos de comercialização, mediante lixiviação periódica do substrato.

Materiais corretivos e nitrogênio na nutrição e na produção de matéria seca de cana-de-açúcar cultivada em vaso

Silicon is a beneficial element that can increase the yield of sugarcane, especially when associated with nitrogen. The aim of this study was to evaluate the effects of silicate slag as a source of Si and its interaction with soil N on nutrition and dry matter production of sugar cane. The treatments consisted of two sources of correctives (limestone and silicate slag) in three doses (one half, once and twice the dose shown to increase V to 60%), two N rates (300 e 600mg.dm(-3)) administered as urea, a control (without corrective and without N), arranged in a completely randomized factorial design 2x3x2+1, with four replicates. Each experimental unit was constituted by a 40dm(3) pot filled with an Oxisol (V= 10% and Si= 10.3mg.dm(-3)), and 4 seedlings (SP79-1011). The nutritional status (Si and N) was evaluated 10 months after planting the sugarcane, as well as the Si content of the soil and shoot dry matter The application of silicate slag increased the content of available Si in soil and the plant. The application of corrective materials improved the nutrition and dry matter production of sugarcane. Nitrogen fertilization was not significant for dry matter production of the sugarcane-plant, regardless of the source of corrective material.

Um sistema para estimação da vazão de gás de poços produzindo por Plunger Lift para vaso separador de teste em plataformas de petróleo

This work intends to analyze the behavior of the gas flow of plunger lift wells producing to well testing separators in offshore production platforms to aim a technical procedure to estimate the gas flow during the slug production period. The motivation for this work appeared from the expectation of some wells equipped with plunger lift method by PETROBRAS in Ubarana sea field located at Rio Grande do Norte State coast where the produced fluids measurement is made in well testing separators at the platform. The oil artificial lift method called plunger lift is used when the available energy of the reservoir is not high enough to overcome all the necessary load losses to lift the oil from the bottom of the well to the surface continuously. This method consists, basically, in one free piston acting as a mechanical interface between the formation gas and the produced liquids, greatly increasing the well s lifting efficiency. A pneumatic control valve is mounted at the flow line to control the cycles. When this valve opens, the plunger starts to move from the bottom to the surface of the well lifting all the oil and gas that are above it until to reach the well test separator where the fluids are measured. The well test separator is used to measure all the volumes produced by the well during a certain period of time called production test. In most cases, the separators are designed to measure stabilized flow, in other words, reasonably constant flow by the use of level and pressure electronic controllers (PLC) and by assumption of a steady pressure inside the separator. With plunger lift wells the liquid and gas flow at the surface are cyclical and unstable what causes the appearance of slugs inside the separator, mainly in the gas phase, because introduce significant errors in the measurement system (e.g.: overrange error). The flow gas analysis proposed in this work is based on two mathematical models used together: i) a plunger lift well model proposed by Baruzzi [1] with later modifications made by Bolonhini [2] to built a plunger lift simulator; ii) a two-phase separator model (gas + liquid) based from a three-phase separator model (gas + oil + water) proposed by Nunes [3]. Based on the models above and with field data collected from the well test separator of PUB-02 platform (Ubarana sea field) it was possible to demonstrate that the output gas flow of the separator can be estimate, with a reasonable precision, from the control signal of the Pressure Control Valve (PCV). Several models of the System Identification Toolbox from MATLAB® were analyzed to evaluate which one better fit to the data collected from the field. For validation of the models, it was used the AIC criterion, as well as a variant of the cross validation criterion. The ARX model performance was the best one to fit to the data and, this way, we decided to evaluate a recursive algorithm (RARX) also with real time data. The results were quite promising that indicating the viability to estimate the output gas flow rate from a plunger lift well producing to a well test separator, with the built-in information of the control signal to the PCV

Projeto de um sistema emulador de escoamentos e vaso de separação primária

The purpose of this study was to develop a pilot plant which the main goal is to emulate a flow peak pressure in a separation vessel. Effect similar that is caused by the production in a slug flow in production wells equipped with the artificial lift method plunger lift. The motivation for its development was the need to test in a plant on a smaller scale, a new technique developed to estimate the gas flow in production wells equipped with plunger lift. To develop it, studies about multiphase flow effects, operation methods of artificial lift in plunger lift wells, industrial instrumentation elements, control valves, vessel sizing separators and measurement systems were done. The methodology used was the definition of process flowcharts, its parameters and how the effects needed would be generated for the success of the experiments. Therefore, control valves, the design and construction of vessels and the acquisition of other equipment used were defined. One of the vessels works as a tank of compressed air that is connected to the separation vessel and generates pulses of gas controlled by a on/off valve. With the emulator system ready, several control experiments were made, being the control of peak flow pressure generation and the flow meter the main experiments, this way, it was confirmed the efficiency of the plant usage in the problem that motivated it. It was concluded that the system is capable of generate effects of flow with peak pressure in a primary separation vessel. Studies such as the estimation of gas flow at the exit of the vessel and several academic studies can be done and tested on a smaller scale and then applied in real plants, avoiding waste of time and money.

Análise de crescimento do crisântemo de vaso, cultivar Puritan, irrigado em diferentes tensões de água em ambiente protegido

A irrigação é prática fundamental para o cultivo de crisântemo, porém seu manejo adequado tem sido negligenciado pelos produtores, resultando em prejuízos no crescimento vegetal e conseqüentes decréscimos na produtividade e na qualidade do produto final. Para melhorar a representatividade dos dados obtidos, o experimento foi desenvolvido na propriedade de um produtor tradicional, no Distrito de Holambra II, município de Paranapanema-SP (23º4'S e 49º00'W). O objetivo principal do trabalho foi identificar a tensão de água no substrato (potencial matricial) que resultasse em melhor crescimento e desenvolvimento do crisântemo em vaso, cultivar Puritan. Os tratamentos foram definidos por seis níveis de tensão de água no substrato: -2, -3, -4,- 6,-10 e -30 kPa. Conclui-se, que a melhor qualidade do crisântemo em vaso foi obtida nos potenciais de água no substrato de -2, -6 e -10 kPa e que a tensão de -30 kPa, embora tenha levado à redução na qualidade comercial do crisântemo, resultou em maior durabilidade pós- colheita.