The secretome of endothelial progenitor cells promotes brain endothelial cell activity through PI3-kinase and MAP-kinase

BACKGROUND Angiogenesis and vascular remodelling are crucial events in tissue repair mechanisms promoted by cell transplantation. Current evidence underscores the importance of the soluble factors secreted by stem cells in tissue regeneration. In the present study we investigated the effects of paracrine factors derived from cultured endothelial progenitor cells (EPC) on rat brain endothelial cell properties and addressed the signaling pathways involved. METHODS Endothelial cells derived from rat brain (rBCEC4) were incubated with EPC-derived conditioned medium (EPC-CM). The angiogenic response of rBCEC4 to EPC-CM was assessed as effect on cell number, migration and tubular network formation. In addition, we have compared the outcome of the in vitro experiments with the effects on capillary sprouting from rat aortic rings. The specific PI3K/AKT inhibitor LY294002 and the MEK/ERK inhibitor PD98059 were used to study the involvement of these two signaling pathways in the transduction of the angiogenic effects of EPC-CM. RESULTS Viable cell number, migration and tubule network formation were significantly augmented upon incubation with EPC-CM. Similar findings were observed for aortic ring outgrowth with significantly longer sprouts. The EPC-CM-induced activities were significantly reduced by the blockage of the PI3K/AKT and MEK/ERK signaling pathways. Similarly to the outcome of the rBCEC4 experiments, inhibition of the PI3K/AKT and MEK/ERK pathways significantly interfered with capillary sprouting induced by EPC-CM. CONCLUSION The present study demonstrates that EPC-derived paracrine factors substantially promote the angiogenic response of brain microvascular endothelial cells. In addition, our findings identified the PI3K/AKT and MEK/ERK pathways to play a central role in mediating these effects.

Effects of shape Rosmarinus officinalis neighbors on resprouting of shape Erica multiflora individuals

To study the effects of competition in Mediterranean shrubland regeneration following disturbance, we used a neighborhood approach to assess the influence of mature Rosmarinus officinalis neighbors on the resprouting of Erica multiflora individuals after clipping. Sprout biomass of target plants 2 years after clipping was regressed against various measures of neighbor abundance within a 2 m radius around target E. multiflora individuals in which all vegetation except R. officinalis had been removed. The largest single influence on the biomass of sprouts produced was the previous biomass of the resprouting plant. The abundance of R. officinalis neighbors had a weak but detectable effect on resprouting of E. multiflora. Abundance of neighbors within 60 cm from target plants was the best predictor of regrowth. At this distance, two simple measures of neighbor abundance within the neighborhood, the number of neighbors and the sum of their heights, were significant in accounting for variation in resprouted biomass. None of the combinations of neighbor variables performed significantly better than single variables. The best models accounted for around 24 percent of the variation in resprout biomass. As in other studies, angular dispersion of neighbors never had a significant effect on performance of target plants. The weak but significant response of resprouting to variation in R. officinalis abundance suggests that the intensity of competition in the experiment was low because of the removal of other species.

Grapholita molesta

La larva de Grapholita molesta (Busck), que produce daños en brotes y frutos de carozo y de pepita, fue criada en condiciones artificiales con dietas a base de granos y otros nutrientes. Se prepararon y evaluaron siete dietas con poroto, garbanzo, arveja, maíz, soja, trigo y avena. Los objetivos del trabajo fueron: 1. Evaluar dietas con granos de cultivares locales. 2. Evaluar las características físico-químicas (pH y sólidos solubles). 3. Evaluar la eficiencia mediante ciclo biológico, grados día, viabilidad, peso de pupas, recuperación de huevo a adulto, fecundidad y adultos esperados. 4. Determinar costos y rendimientos. El análisis de los parámetros biológicos y económicos indicó que las dietas de menor costo y mayor rendimiento fueron, en orden decreciente, poroto y garbanzo, pero la primera además presentó características físicoquímicas similares a la dieta preferencial de la larva salvaje: los brotes de duraznero.

Pollen analysis, particle size analysis, end-member modelling and x-ray fluorescence measurements from sediment cores off Cape Ghir, Morocco

Understanding past human-climate-environment interactions is essential for assessing the vulnerability of landscapes and ecosystems to future climate change. This is particularly important in southern Morocco where the current vegetation is impacted by pastoralism, and the region is highly sensitive to climate variability. Here, we present a 2000-year record of vegetation, sedimentation rate, XRF chemical element intensities, and particle size from two decadal-resolved, marine sediment cores, raised from offshore Cape Ghir, southern Morocco. The results show that between 650 and 850 AD the sedimentation rate increased dramatically from 100 cm/1000 years to 300 cm/1000 years, and the Fe/Ca and pollen flux doubled, together indicating higher inputs of terrestrial sediment. Particle size measurements and end-member modelling suggest increased fluvial transport of the sediment. Beginning at 650 AD pollen levels from Cichorioideae species show a sharp rise from 10% to 20%. Pollen from Atemisia and Plantago, also increase from this time. Deciduous oak pollen percentages show a decline, whereas those of evergreen oak barely change. The abrupt increase in terrestrial/fluvial input from 650 to 850 AD occurs, within the age uncertainty, of the arrival of Islam (Islamisation) in Morocco at around 700 AD. Historical evidence suggests Islamisation led to population increase and development of southern Morocco, including expanded pastoralism, deforestation and agriculture. Livestock pressure may have changed the vegetation structure, accounting for the increase in pollen from Cichorioideae, Plantago, and Artemisia, which include many weedy species. Goats in particular may have played a dominant role as agents of erosion, and intense browsing may have led to the decline in deciduous oak; evergreen oak is more likely to survive as it re-sprouts more vigorously after browsing. From 850 AD to present sedimentation rates, Fe/Ca ratios and fluvial discharge remain stable, whereas pollen results suggest continued degradation. Pollen results from the past 150 years suggest expanded cultivation of olives and the native argan tree, and the introduction of Australian eucalyptus trees. The rapidly increasing population in southern Morocco is causing continued pressure to expand pastoralism and agriculture. The history of land degradation presented here suggests that the vegetation in southern Morocco may have been degraded for a longer period than previously thought and may be particularly sensitive to further land use changes. These results should be included in land management strategies for southern Morocco.

Vascular endothelial growth factor C induces angiogenesis in vivo

Vascular endothelial growth factor C (VEGF-C) recently has been described to be a relatively specific growth factor for the lymphatic vascular system. Here we report that ectopic application of recombinant VEGF-C also has potent angiogenic effects in vivo. VEGF-C is sufficiently potent to stimulate neovascularization from limbal vessels in the mouse cornea. Similar to VEGF, the angiogenic response of corneas induced by VEGF-C is intensive, with a high density of new capillaries. However, the outgrowth of microvessels stimulated by VEGF-C was significantly longer than that induced by VEGF. In the developing embryo, VEGF-C was able to induce branch sprouts from the established blood vessels. VEGF-C also induced an elongated, spindle-like cell shape change and actin reorganization in both VEGF receptor (VEGFR)-2 and VEGFR-3-overexpressing endothelial cells, but not in VEGFR-1-expressing cells. Further, both VEGFR-2 and VEGFR-3 could mediate proliferative and chemotactic responses in endothelial cells on VEGF-C stimulation. Thus, VEGF-C may regulate physiological angiogenesis and participate in the development and progression of angiogenic diseases in addition to lymphangiogenesis.

Synaptic sprouting increases the uptake capacities of motoneurons in amyotrophic lateral sclerosis mice

Using adenoviruses encoding reporter genes as retrograde tracers, we assessed the capacity of motoneurons to take up and retrogradely transport adenoviral particles injected into the muscles of transgenic mice expressing the G93A human superoxide dismutase mutation, a model of amyotrophic lateral sclerosis. Surprisingly, transgene expression in the motoneurons was significantly higher in symptomatic mice than in control or presymptomatic mice. Using botulinum toxin to induce nerve sprouting at neuromuscular junctions, we showed that the unexpectedly high level of motoneurons retrograde transduction results, at least in part, from newly acquired uptake properties of the sprouts. These findings demonstrate the remarkable uptake properties of amyotrophic lateral sclerosis motoneurons in response to denervation and the rationale of using intramuscular injections of adenoviruses to overexpress therapeutic proteins in motor neuron diseases.

Mini-toletes de cana-de-açúcar: gemas, biorreguladores, adubação nitrogenada e déficit hídrico

A cana-de-açúcar é uma espécie amplamente cultivada em regiões tropicais e subtropicais. Sua propagação é realizada através do plantio de porções caulinares contendo uma média de três gemas. Tal prática requer grande quantidade de material vegetal, o que reduz o ganho dos produtores. Adicionalmente, a utilização de grande quantidade de material vegetal para o plantio dificulta algumas práticas em relação ao manejo da cultura, como transporte e armazenamento. A utilização de mini-toletes, contendo uma única gema, representa uma alternativa ao plantio convencional. Existem limitações impostas à utilização de mini-toletes, relacionadas à baixa disponibilidade de reservas de nutrientes e de água, devido ao reduzido tamanho dos toletes. O presente trabalho teve o objetivo de avaliar o vigor e o desenvolvimento de plantas de cana-de-açúcar provenientes de mini-toletes. No primeiro experimento, comparou-se plantas provenientes de diferentes tipos de propágulos e gemas. Foram avaliados o número de brotações, a porcentagem de brotações, a altura das plantas e as massas de folhas e raízes. No segundo experimento, avaliou-se o efeito da aplicação de biorreguladores em mini-toletes provenientes de gemas apicais e basais. Foram realizadas as determinações do número de brotações, da altura das plantas, da área foliar e das massas secas de folhas e colmos. No terceiro experimento, avaliou-se a aplicação de ureia como fonte de adubação nitrogenada e tiametoxam, um inseticida sistêmico com ação bioativadora, no desenvolvimento de plantas originárias de mini-toletes. Realizaram-se as seguintes determinações: número de brotações, altura das plantas, área foliar e massas secas de colmos, folhas e raízes. No quarto experimento, plantas de cana-de-açúcar originárias de mini-toletes tratados com agroquímicos foram submetidas ao déficit hídrico. Foram avaliadas a altura das plantas, a área foliar e as massas de raiz, folha e caule. Foi possível concluir que plantas provenientes de gemas superiores e de toletes contendo três e duas gemas apresentaram um melhor desenvolvimento. De maneira geral, os resultados indicaram que a cana-de-açúcar não responde de maneira evidente ao uso de reguladores vegetais em mini-toletes. A utilização de ureia aumenta o desenvolvimento de plantas originárias de mini-toletes. Em conjunto com diferentes doses de ureia, a utilização de tiametoxam incrementa aspectos do desenvolvimento da cana-de-açúcar. Adicionalmente, foi possível concluir que a aplicação de agroquímicos em mini-toletes alivia os efeitos negativos do déficit hídrico no desenvolvimento radicular. A partir dos resultados obtidos no presente trabalho, foi possível concluir que a utilização de ureia, tiametoxam e agroquímicos melhora o desenvolvimento de plantas de cana-de-açúcar originárias de mini-toletes.

Dynamics of motor nerve terminal remodeling unveiled using SNARE-cleaving botulinum toxins: the extent and duration are dictated by the sites of SNAP-25 truncation

Nerve sprouts emerge from motor nerve terminals following blockade of exo-endocytosis for more than 3 days by botulinum neurotoxin (BoNT), and form functional synapses, albeit temporary. Upon restoration of synaptic activity to the parent terminal 7 and 90 days after exposure to BoNT/F or A respectively, a concomitant retraction of the outgrowths was observed. BoNT/E caused short-term neuroparalysis, and dramatically accelerated the recovery of BoNT/A-paralyzed muscle by further truncation of SNAP-25 and its replenishment with functional full-length SNARE. The removal of 9 C-terminal residues from SNAP-25 by BoNT/A leads to persistence of the inhibitory product due to the formation of a nonproductive SNARE complex(es) at release sites, whereas deletion of a further 17 amino acids permits replenishment and a speedy recovery. (C) 2003 Elsevier Science (USA). All rights reserved.

Vascular endothelial growth factor-B and retinal vascular development in the mouse

Purpose: Vascular endothelial growth factor-A (VEGF-A) is crucial to retinal vascular growth, both normal and pathological. VEGF-B, recently characterized, is reported to be expressed in retinal tissues, but the importance of VEGF-B to retinal vascular development remained unknown. The aim of this study was to analyse retinal vascular growth in the Vegfb (-/-) knockout mouse. Methods: Retinal vascular growth was measured in Vegfb (-/-) knockout mice raised under normal conditions, and Vegfb (-/-) knockout mice with an oxygen-induced proliferative retinopathy. Wild type Vegfb (+/+) mice served as controls. Vessels were perfused with ink and retinal flatmounts secondarily labelled with FITC-lectin (BS-1, Griffonia simplicifolia ). Area and diameter of retinal growth and retinal vascular growth were recorded over days 0-20, and capillary density and mean diameter recorded from day 17 pups. Results: A variety of techniques confirmed that Vegfb (+/+) mice expressed VEGF-B and that VEGF-B expression was absent in Vegfb (-/-) mice. Vegfb (-/-) mice raised in room air showed no significant differences from Vegfb (+/+) controls. No differences were found in oxygen-induced retinopathy between Vegfb (-/-) and Vegfb (+/+) pups in either the extent of the initial oxygen-induced ablation, or in the regrowth of retinal vessels or vitreal (neovascular) sprouts; vitreal sprouts are important markers of the abnormal proliferative response, and are maximally expressed on day 17 in this model of oxygen-induced retinopathy. Conclusions: These results indicate that a lack of VEGF-B does not significantly affect development of the retinal vasculature under normal conditions, nor does it appear to affect the proliferative retinal responses seen in oxygen-induced retinopathy.

Development of a food composition database for the estimation of dietary intakes of glucosinolates, the biologically active constituents of cruciferous vegetables

Evidence indicates that cruciferous vegetables are protective against a range of cancers with glucosinolates and their breakdown products considered the biologically active constituents. To date, epidemiological studies have not investigated the intakes of these constituents due to a lack of food composition databases. The aim of the present study was to develop a database for the glucosinolate content of cruciferous vegetables that can be used to quantify dietary exposure for use in epidemiological studies of diet-disease relationships. Published food composition data sources for the glucosinolate content of cruciferous vegetables were identified and assessed for data quality using established criteria. Adequate data for the total glucosinolate content were available from eighteen published studies providing 140 estimates for forty-two items. The highest glucosinolate values were for cress (389 mg/100 g) while the lowest values were for Pe-tsai chinese cabbage (20 mg/100 g). There is considerable variation in the values reported for the same vegetable by different studies, with a median difference between the minimum and maximum values of 5.8-fold. Limited analysis of cooked cruciferous vegetables has been conducted; however, the available data show that average losses during cooking are approximately 36 %. This is the first attempt to collate the available literature on the glucosinolate content of cruciferous vegetables. These data will allow quantification of intakes of the glucosinolates, which can be used in epidemiological studies to investigate the role of cruciferous vegetables in cancer aetiology and prevention.

Effect of nitrogen supply on leaf appearance, leaf growth, leaf nitrogen economy and photosynthetic capacity in maize (Zea mays L.)

Leaf area growth and nitrogen concentration per unit leaf area, N-a (g m(-2) N) are two options plants can use to adapt to nitrogen limitation. Previous work indicated that potato (Solanum tuberosum L.) adapts the size of leaves to maintain Na and photosynthetic capacity per unit leaf area. This paper reports on the effect of N limitation on leaf area production and photosynthetic capacity in maize, a C4 cereal. Maize was grown in two experiments in pots in glasshouses with three (0.84-6.0 g N pot(-1)) and five rates (0.5-6.0 g pot(-1)) of N. Leaf tip and ligule appearance were monitored and final individual leaf area was determined. Changes with leaf age in leaf area, leaf N content and light-saturated photosynthetic capacity, P a,, were measured on two leaves per plant in each experiment. The final area of the largest leaf and total plant leaf area differed by 16 and 29% from the lowest to highest N supply, but leaf appearance rate and the duration of leaf expansion were unaffected. The N concentration of expanding leaves (N-a or %N in dry matter) differed by at least a factor 2 from the lowest to highest N supply. A hyperbolic function described the relation between P-max and N-a. The results confirm the 'maize strategy': leaf N content, photosynthetic capacity, and ultimately radiation use efficiency is more sensitive to nitrogen limitation than are leaf area expansion and light interception. The generality of the findings is discussed and it is suggested that at canopy level species showing the 'potato strategy' can be recognized from little effect of nitrogen supply on radiation use efficiency, while the reverse is true for species showing the 'maize strategy' for adaptation to N limitation. (c) 2004 Elsevier B.V. All rights reserved.

Flt1 acts as a negative regulator of tip cell formation and branching morphogenesis in the zebrafish embryo

Endothelial tip cells guide angiogenic sprouts by exploring the local environment for guidance cues such as vascular endothelial growth factor (VegfA). Here we present Flt1 (Vegf receptor 1) loss- and gain-of-function data in zebrafish showing that Flt1 regulates tip cell formation and arterial branching morphogenesis. Zebrafish embryos expressed soluble Flt1 (sFlt1) and membrane-bound Flt1 (mFlt1). In Tg(flt1(BAC):yfp) × Tg(kdrl:ras-cherry)(s916) embryos, flt1:yfp was expressed in tip, stalk and base cells of segmental artery sprouts and overlapped with kdrl:cherry expression in these domains. flt1 morphants showed increased tip cell numbers, enhanced angiogenic behavior and hyperbranching of segmental artery sprouts. The additional arterial branches developed into functional vessels carrying blood flow. In support of a functional role for the extracellular VEGF-binding domain of Flt1, overexpression of sflt1 or mflt1 rescued aberrant branching in flt1 morphants, and overexpression of sflt1 or mflt1 in controls resulted in short arterial sprouts with reduced numbers of filopodia. flt1 morphants showed reduced expression of Notch receptors and of the Notch downstream target efnb2a, and ectopic expression of flt4 in arteries, consistent with loss of Notch signaling. Conditional overexpression of the notch1a intracellular cleaved domain in flt1 morphants restored segmental artery patterning. The developing nervous system of the trunk contributed to the distribution of Flt1, and the loss of flt1 affected neurons. Thus, Flt1 acts in a Notch-dependent manner as a negative regulator of tip cell differentiation and branching. Flt1 distribution may be fine-tuned, involving interactions with the developing nervous system.

Degradation of Biofumigant Isothiocyanates and Allyl Glucosinolate in Soil and Their Effects on the Microbial Community Composition

Brassicales species rich in glucosinolates are used for biofumigation, a process based on releasing enzymatically toxic isothiocyanates into the soil. These hydrolysis products are volatile and often reactive compounds. Moreover, glucosinolates can be degraded also without the presence of the hydrolytic enzyme myrosinase which might contribute to bioactive effects. Thus, in the present study the stability of Brassicaceae plant-derived and pure glucosinolates hydrolysis products was studied using three different soils ( model biofumigation). In addition, the degradation of pure 2-propenyl glucosinolate was investigated with special regard to the formation of volatile breakdown products. Finally, the influence of pure glucosinolate degradation on the bacterial community composition was evaluated using denaturing gradient gel electrophoresis of 16S rRNA gene amplified from total community DNA. The model biofumigation study revealed that the structure of the hydrolysis products had a significant impact on their stability in the soil but not the soil type. Following the degradation of pure 2-propenyl glucosinolate in the soils, the nitrile as well as the isothiocyanate can be the main degradation products, depending on the soil type. Furthermore, the degradation was shown to be both chemically as well as biologically mediated as autoclaving reduced degradation. The nitrile was the major product of the chemical degradation and its formation increased with iron content of the soil. Additionally, the bacterial community composition was significantly affected by adding pure 2-propenyl glucosinolate, the effect being more pronounced than in treatments with myrosinase added to the glucosinolate. Therefore, glucosinolates can have a greater effect on soil bacterial community composition than their hydrolysis products.

Optimising plant bed performance through adaptive research with the Australian sweetpotato industry

A key driver of Australian sweetpotato productivity improvements and consumer demand has been industry adoption of disease-free planting material systems. On a farm isolated from main Australian sweetpotato areas, virus-free germplasm is annually multiplied, with subsequent 'pathogen-tested' (PT) sweetpotato roots shipped to commercial Australian sweetpotato growers. They in turn plant their PT roots into specially designated plant beds, commencing in late winter. From these beds, they cut sprouts as the basis for their commercial fields. Along with other intense agronomic practices, this system enables Australian producers to achieve worldRSQUOs highest commercial yields (per hectare) of premium sweetpotatoes. Their industry organisation, ASPG (Australian Sweetpotato Growers Inc.), has identified productivity of mother plant beds as a key driver of crop performance. Growers and scientists are currently collaborating to investigate issues such as catastrophic plant beds losses; optimisation of irrigation and nutrient addition; rapidity and uniformity of initial plant bed harvests; optimal plant bed harvest techniques; virus re-infection of plant beds; and practical longevity of plant beds. A survey of 50 sweetpotato growers in Queensland and New South Wales identified a substantial diversity in current plant bed systems, apparently influenced by growing district, scale of operation, time of planting, and machinery/labour availability. Growers identified key areas for plant bed research as: optimising the size and grading specifications of PT roots supplied for the plant beds; change in sprout density, vigour and performance through sequential cuttings of the plant bed; optimal height above ground level to cut sprouts to maximise commercial crop and plant bed performance; and use of structures and soil amendments in plant bed systems. Our ongoing multi-disciplinary research program integrates detailed agronomic experiments, grower adaptive learning sites, product quality and consumer research, to enhance industry capacity for inspired innovation and commercial, sustainable practice change.

Caracterização de linhagens de soja Glycine max (L.) Merrill para produção de brotos.

2016