Abscisic acid and auxin accumulation in Catasetum fimbriatum roots growing in vitro with high sucrose and mannitol content

Endogenous contents of indolyl-3-acetic acid (IAA) and abscisic acid (ABA) were quantified in excised roots of Catasetum fimbriatum (Orchidaceae) cultured in vitro on solidified Vacin and Went medium with 1, 2, 4, 6, 8 and 10 % sucrose, as well as 2 % sucrose plus mannitol. Maximum root growth was observed in media with 4 % sucrose and 2 % sucrose plus 2.2 % mannitol, suggesting that a moderate water or osmotic stress promotes orchid root growth. Contents of both ABA and IAA increased in parallel to increasing sucrose concentration and a correlation between root elongation and the ABA/IAA ratio was observed. Incubating isolated C. fimbriatum roots with radiolabeled tryptophan, we showed an accumulation of IAA and its conjugates.

Desiccation tolerance in Pleurostima purpurea (Velloziaceae)

Previous works suggested that Pleurostima purpurea (Velloziaceae-Barbacenioideae) shows a remarkable capacity to endure desiccation of its vegetative tissues. P. purpurea occurs in monocotyledons mats on soil islands in the Pao de Acucar (Sugar Loaf) one of the most recognizable rock outcrops of the world, in Rio de Janeiro, southeastern Brazil. Mats of P. purpurea occur in cliffs by the sea some meters above the tidal zone. Although living in rock outcrops almost devoid of any soil cover, P. purpurea seems to occur preferably on less exposed rock faces and slightly shady sites. Usually, less extreme adaptations to drought would be expected in plants with the habitat preference of P. purpurea. Relying on this observation, we argue if a combination of different strategies of dealing with low water availability can be found in P. purpurea as on other desiccation tolerant angiosperms. This study aims to examine the occurrence of desiccation tolerant behavior in P. purpurea together with the expression of drought avoidance mechanisms during dehydration progression. For this, it was analyzed the gas exchanges, leaf pigments and relative leaf water content during desiccation and rehydration of cultivated mature individuals. P. purpurea behaved like typical drought avoiders under moderated drought condition with stomatal closure occurring around a relative leaf water content up to 90%. During this process, it was observed a delay in the leaf relative water content (RWC(leaf)) decrease comparing to the plant-soil relative water content (RWC(plant-soil)). As soil dehydration worsened, gas exchanges restrictions progressed until a lack of activity which characterizes anabiosis. The loss of chlorophyll occurs before the end of total dehydration, characterizing the presence of poikilochlorophylly. The chlorophyll degradation follows the RWC(leaf) decrease, which achieved the minimum average value of 17% without incurring in leaf abscission. The chlorophyll re-synthesis seems to start well after the full rehydration of the leaf. During all of this process, carotenoid content remained stable. These results are coherent with a combination of drought avoidance and desiccation tolerance in P. purpurea which seems to be coherent with the amplitude of water availability in the rock outcrop habitat where it occurs, suggesting that the periods of water availability are sufficiently long for the success of the costly desiccation tolerant behavior but too short to make a typical drought avoider species win the competition for exploring the rock outcrop substrate where P. purpurea occurs.

Habitat Split as a Cause of Local Population Declines of Amphibians with Aquatic Larvae

Most amphibian species have biphasic life histories and undergo an ontogenetic shift from aquatic to terrestrial habitats. In deforested landscapes, streams and forest fragments are frequently disjunct, jeopardizing the life cycle of forest-associated amphibians with aquatic larvae. We tested the impact of habitat split-defined as human-induced disconnection between habitats used by different life-history stages of a species-on four forest-associated amphibian species in a severely fragmented landscape of the Brazilian Atlantic Forest. We surveyed amphibians in forest fragments with and without streams (referred to as wet and dry fragments, respectively), including the adjacent grass-field matrix. Our comparison of capture rates in dry fragments and nearby streams in the matrix allowed us to evaluate the number of individuals that engaged in high-risk migrations through nonforested habitats. Adult amphibians moved from dry fragments to matrix streams at the beginning of the rainy season, reproduced, and returned at the end of the breeding period. Juveniles of the year moved to dry fragments along with adults. These risky reproductive migrations through nonforested habitats that expose individuals to dehydration, predation, and other hazards may cause population declines in dry fragments. Indeed, capture rates were significantly lower in dry fragments compared with wet fragments. Declining amphibians would strongly benefit from investments in the conservation and restoration of riparian vegetation and corridors linking breeding and nonbreeding areas.

Effect of Mussel`s Gender and Size on a Stress Response Biomarker

In mussels, stress signals such as heat, osmotic shock and hypoxia lead to the activation of the phosphorylated p38 mitogen activated protein kinase (pp38-MAPK). This stress activated protein has been efficiently used as a biomarker to several natural and anthropogenic stresses. However, what has not been tested is whether differences in gender or size can affect the response of this biomarker. The present study tested whether there was variation in the expression of pp38-MAPK in mussels Perna perna of different gender and size classes when exposed to natural stress conditions, such as air exposure. The results show that gender does not affect the expression of pp38-MAPK. However, size does have an effect, where mussels smaller than 6.5 cm displayed significantly (p < 0.05) lower levels of pp38-MAPK when compared to those larger than 7 cm. Mussels are one of the most used bioindicator species and the use of biomarkers to determine the health status of an ecosystem has been greatly increasing over the years. The present study highlights the importance of using mussels of similar size classes when performing experiments using stress-related biomarkers.

Role of spinal microglia in myositis-induced central sensitisation: An immunohistochemical and behavioural study in rats

There is increasing evidence that spinal glial cells play an important role in chronic pain states. However, so far no data on the role of microglia in muscle pain are available. The aim of the present study was to investigate the involvement of spinal microglial cells in chronic muscle pain. In a rat model of chronic muscle inflammation (injection of complete Freunds adjuvant into the gastrocnemius-soleus muscle) alterations of microglia were visualized with quantitative OX-42 immunohistochemistry in the dorsal horn of the segments L4 and L5 12 days after induction of inflammation. In behavioural experiments the influence of chronic intrathecally applied minocycline - a specific microglia inhibitor - or an antibody against tumour necrosis factor-alpha (TNF-alpha: a cytokine released from microglia) on pain-related behaviour was investigated after 1, 3, 6, and 12 days. The immunhistochemical data show that in the deep laminae of the spinal dorsal horn microglial cells reacted with morphological changes to the muscle inflammation. Following inflammation, the mean boundary length surrounding the OX-42 immunostained area was significantly shorter. This indicates that microglial cells were activated by the myositis and withdrew their processes. Chronic intrathecal administration of minocycline or anti TNF-alpha with an osmotic mini-pump largely normalised the inflammation-induced changes in spontaneous exploratory behaviour and attenuated the hypersensitivity to mechanical stimulation. Both the immunohistochemical and behavioural data show that spinal microglial cells are involved in nociceptive processes in the cause of a chronic muscle inflammation. (C) 2008 European Federation of International Association for the Study of Pain Chapters. Published by Elsevier Ltd. All rights reserved.

Cloning and characterization of a zebrafish homologue of human AQP1: a bifunctional water and gas channel

Chen LM, Zhao J, Musa-Aziz R, Pelletier MF, Drummond IA, Boron WF. Cloning and characterization of a zebrafish homologue of human AQP1: a bifunctional water and gas channel. Am J Physiol Regul Integr Comp Physiol 299: R1163-R1174, 2010. First published August 25, 2010; doi:10.1152/ajpregu.00319.2010.-The mammalian aquaporins AQP1, AQP4, and AQP5 have been shown to function not only as water channels but also as gas channels. Zebrafish have two genes encoding an AQP1 homologue, aqp1a and aqp1b. In the present study, we cloned the cDNA that encodes the zebrafish protein Aqp1a from the 72-h postfertilization (hpf) embryo of Danio rerio, as well as from the swim bladder of the adult. The deduced amino-acid sequence of aqp1a consists of 260 amino acids and is 59% identical to human AQP1. By analyzing the genomic DNA sequence, we identified four exons in the aqp1a gene. By in situ hybridization, aqp1a is expressed transiently in the developing vasculature and in erythrocytes from 16 to 48 h of development. Later, at 72 hpf, aqp1a is expressed in dermal ionocytes and in the swim bladder. Western blot analysis of adult tissues reveals that Aqp1a is most highly expressed in the eye and swim bladder. Xenopus oocytes expressing aqp1a have a channel-dependent (*) osmotic water permeability (P(f)*) that is indistinguishable from that of human AQP1. On the basis of the magnitude of the transient change in surface pH (Delta pHS) that were recorded as the oocytes were exposed to either CO(2) or NH(3), we conclude that zebrafish Aqp1a is permeable to both CO(2) and NH(3). The ratio (Delta pHS*)CO2/P(f)* is about half that of human AQP1, and the ratio (Delta pHS*)NH3/P(f)* is about one-quarter that of human AQP1. Thus, compared with human AQP1, zebrafish Aqp1a has about twice the selectivity for CO(2) over NH(3).

Lonomia obliqua (Lepidoptera, Saturniidae) caterpillar bristle extract induces direct lysis by cleaving erythrocyte membrane glycoproteins

Lonomia obliqua caterpillar bristle extract induces hemolysis in vitro on washed human and rat erythrocytes, in either the absence or presence of exogenous lecithin. In the former condition, phospholipases A(2) are key enzymes involved in hemolysis. However, the mechanism whereby this extract causes direct hemolysis is not known. Thus, the aim of this study was to investigate the hemolytic mechanism of the crude extract of the caterpillar L obliqua on human erythrocytes in the absence of lecithin. The extract significantly increased the erythrocyte osmotic fragility and promoted the removal of glycophorins A and C, and band 3 from the erythrocyte membrane. The use of Ca(2+) and Mg(2+) ions significantly potentiated glycoprotein removal, remarkably of erythrocyte band 3. The composition of fatty acids was analyzed by HPLC in both L obliqua caterpillar bristle extract and human erythrocyte membranes incubated with the extract. The levels of unsaturated fatty acids were remarkably augmented in erythrocytes incubated with the extract than in control erythrocytes, modifying thereby the saturated/unsaturated fatty acid ratio. Altogether, evidence is provided here that the interplay of at least three mechanisms of action accounts for the direct activity of the bristle extract on erythrocyte membrane, leading to hemolysis: the removal of glycoproteins and band 3; the insertion of fatty acids; and the action of phospholipases. Such mechanisms might affect erythrocyte flexibility and deformability, which may induce hemolysis by increasing erythrocyte fragility. However, whether the direct hemolytic activity of L obliqua caterpillar is the major cause of intravascular hemolysis during envenomation still needs further investigation. (C) 2010 Elsevier Ltd. All rights reserved.

Decreased cGMP Level Contributes to Increased Contraction in Arteries From Hypertensive Rats Role of Phosphodiesterase 1

Recent evidence suggests that angiotensin II (Ang II) upregulates phosphodiesterase (PDE) 1A expression. We hypothesized that Ang II augmented PDE1 activation, decreasing the bioavailability of cyclic guanosine 3` 5`-monophosphate (cGMP), and contributing to increased vascular contractility. Male Sprague-Dawley rats received mini-osmotic pumps with Ang II (60 ng.min(-1)) or saline for 14 days. Phenylephrine (PE)-induced contractions were increased in aorta (E(max)168%+/- 8% vs 136%+/- 4%) and small mesenteric arteries (SMA; E(max)170%+/- 6% vs 143%+/- 3%) from Ang II-infused rats compared to control. PDE1 inhibition with vinpocetine (10 mu mol/L) reduced PE-induced contraction in aortas from Ang II rats (E(max)94%+/- 12%) but not in controls (154%+/- 7%). Vinpocetine decreased the sensitivity to PE in SMA from Ang II rats compared to vehicle (-log of half maximal effective concentration 5.1 +/- 0.1 vs 5.9 +/- 0.06), but not in controls (6.0 +/- 0.03 vs 6.1 +/- 0.04). Sildenafil (10 mu mol/L), a PDE5 inhibitor, reduced PE-induced maximal contraction similarly in Ang II and control rats. Arteries were contracted with PE (1 mu mol/L), and concentration-dependent relaxation to vinpocetine and sildenafil was evaluated. Aortas from Ang II rats displayed increased relaxation to vinpocetine compared to control (E(max)82%+/- 12% vs 445 +/- 5%). SMA from Ang II rats showed greater sensitivity during vinpocetine-induced relaxation compared to control (-log of half maximal effective concentration 6.1 +/- 0.3 vs 5.3 +/- 0.1). No differences in sildenafil-induced relaxation were observed. PDE1A and PDE1C expressions in aorta and PDE1A expression in SMA were increased in Ang II rats. cGMP production, which is decreased in arteries from Ang II rats, was restored after PDE1 blockade. We conclude that PDE1 activation reduces cGMP bioavailability in arteries from Ang II, contributing to increased contractile responsiveness. (Hypertension. 2011;57[part 2]:655-663.)

Gamma-linolenic acid inhibits both tumour cell cycle progression and angiogenesis in the orthotopic C6 glioma model through changes in VEGF, Flt1, ERK1/2, MMP2, cyclin D1, pRb, p53 and p27 protein expression

Background: Gamma-linolenic acid is a known inhibitor of tumour cell proliferation and migration in both in vitro and in vivo conditions. The aim of the present study was to determine the mechanisms by which gamma-linolenic acid (GLA) osmotic pump infusion alters glioma cell proliferation, and whether it affects cell cycle control and angiogenesis in the C6 glioma in vivo. Methods: Established C6 rat gliomas were treated for 14 days with 5 mM GLA in CSF or CSF alone. Tumour size was estimated, microvessel density (MVD) counted and protein and mRNA expression measured by immunohistochemistry, western blotting and RT-PCR. Results: GLA caused a significant decrease in tumour size (75 +/- 8.8%) and reduced MVD by 44 +/- 5.4%. These changes were associated with reduced expression of vascular endothelial growth factor (VEGF) (71 +/- 16%) and the VEGF receptor Flt1 (57 +/- 5.8%) but not Flk1. Expression of ERK1/2 was also reduced by 27 +/- 7.7% and 31 +/- 8.7% respectively. mRNA expression of matrix metalloproteinase-2 (MMP2) was reduced by 35 +/- 6.8% and zymography showed MMP2 proteolytic activity was reduced by 32 +/- 8.5%. GLA altered the expression of several proteins involved in cell cycle control. pRb protein expression was decreased (62 +/- 18%) while E2F1 remained unchanged. Cyclin D1 protein expression was increased by 42 +/- 12% in the presence of GLA. The cyclin dependent kinase inhibitors p21 and p27 responded differently to GLA, p27 expression was increased (27 +/- 7.3%) while p21 remained unchanged. The expression of p53 was increased (44 +/- 16%) by GLA. Finally, the BrdU incorporation studies found a significant inhibition (32 +/- 11%) of BrdU incorporation into the tumour in vivo. Conclusion: Overall the findings reported in the present study lend further support to the potential of GLA as an inhibitor of glioma cell proliferation in vivo and show it has direct effects upon cell cycle control and angiogenesis. These effects involve changes in protein expression of VEGF, Flt1, ERK1, ERK2, MMP2, Cyclin D1, pRb, p53 and p27. Combination therapy using drugs with other, complementary targets and GLA could lead to gains in treatment efficacy in this notoriously difficult to treat tumour.

Confinement-induced phase transition in a DNA-lipid hydrated complex

We study the effect of the soft confinement by fluid lipid bilayers on the spatial organisation of DNA molecules in a DNA-zwitterionic lipid hydrated lamellar complex. The confinement is increased by dehydrating the complex in a controlled way, which leads to a decrease of the water channel thickness separating the periodically stacked bilayers. Using grazing-incidence small-angle X-ray scattering on an oriented thin film, we probe in situ as dehydration proceeds the structure of the DNA-lipid complex. A structural phase transition is evidenced, where an apparently disordered phase of DNA rods embedded within the one-dimensionally ordered lipid lamellar phase observed at high hydration is replaced by a 2D hexagonal structure of DNA molecules intercalated between the lipid bilayers. Copyright (C) EPLA, 2010

Thermoluminescence, infrared reflectivity and electron paramagnetic resonance properties of hemimorphite

Silicate mineral hemimorphite has been investigated concerning its TL, IR and EPR properties. A broad TL peak around 180 degrees C and a weaker and narrower peak around 360 degrees C were found in a sample annealed at 600 degrees C for I h and then irradiated. The deconvolution using the CGCD method revealed peaks around 132, 169, 222 and 367 degrees C. The reflectivity measurements showed several bands in the NIR region due to H(2)O, OH and Al-OH complexes. No band was observed in the visible region. The thermal treatments were carried out from similar to 110 to 940 degrees C and dehydration was observed, first causing a diminishing optical absorption in general and the disappearance of water and hydroxyl absorption bands. The EPR spectrum of natural hemimorphite, presented Cu(2+) signals at g = 2.4 and g = 2.1 plus E(1)` signal superposed to Fe(3+) signal around g = 2.0. (C) 2008 Elsevier Ltd. All rights reserved.

Phase change and pyroelectricity in natural hemimorphite

The phase change of a natural hemimorphite sample from Minas Gerais (Brazil) was investigated by two X-ray diffraction (XRD) methods and by near-infrared reflectance spectroscopy. Applying successive thermal treatments, the crystal structure undergoes two orientation conversions. The first one occurs at about 550 degrees C, and it was revealed by the Laue method. Below 500 degrees C, the water molecules were partially expelled without changing the crystal structure. A fact that supports this statement is the sequential disappearance of the water bands at 1400 and 1900 nm by thermal treatment. The second conversion takes place below 939 degrees C. Moreover, at 972 degrees C a phase change to the willemite mineral (alpha-Zn(2)SiO(4)) has been observed. This last conversion was confirmed by the power XRD. In addition, natural hemimorphite displayed a high pyroelectricity, which is related both to the absence of inverse centre and to the presence of molecular water and hydroxyl groups in the crystal structure.

High T/P evolution and metamorphic ages of the migmatitic basement of northern Sierras Pampeanas, Argentina: Characterization of a mid-crustal segment of the Famatinian belt

New petrologic, thermobarometric and U-Pb monazite geochronologic information allowed to resolve the metamorphic evolution of a high temperature mid-crustal segment of an ancient subduction-related orogen. The EI Portezuelo Metamorphic-Igneous Complex, in the northern Sierras Pampeanas, is mainly composed of migmatites that evolved from amphibolite to granulite metamorphic facies, reaching thermal peak conditions of 670-820 degrees C and 4.5-5.3 kbar. The petrographic study combined with conventional and pseudosection thermobarometry led to deducing a short prograde metamorphic evolution within migmatite blocks. The garnet-absent migmatites represent amphibolite-facies rocks, whereas the cordierite-garnet-K-feldspar-sillimanite migmatites represent higher metamorphic grade rocks. U-Pb geochronology on monazite grains within leucosome record the time of migmatization between approximate to 477 and 470 Ma. Thus, the El Portezuelo Metamorphic-Igneous Complex is an example of exhumed Early Ordovician anatectic middle crust of the Famatinian mobile belt. Homogeneous exposure of similar paleo-depths throughout the Famatinian back-arc and isobaric cooling paths suggest slow exhumation and consequent longstanding crustal residence at high temperatures. High thermal gradients uniformly distributed in the Famatinian back-arc can be explained by shallow convection of a low-viscosity asthenosphere promoted by subducting-slab dehydration. (C) 2011 Elsevier Ltd. All rights reserved.

The link between partial melting, granitization and granulite development in central Ribeira Fold Belt, SE Brazil: New evidence from elemental and Sr-Nd isotopic geochemistry

Elemental and Sr-Nd isotopic data on metatexites, diatexites, orthogneisses and charnockites from the central Ribeira Fold Belt indicate that they are LILE-enriched weakly peraluminous granodiorites. Harker and Th-Hf-La correlation trends suggest that these rocks represent a co-genetic sequence, whereas variations on CaO, MnO, Y and HREE for charnockites can be explained by garnet consumption during granulitic metamorphism. Similar REE patterns and isotopic results of epsilon(565)(Nd) = -5.4 to -7.3 and (87)Sr/(86)Sr(565) = 0.706-0.711 for metatexites, diatexites, orthogneisses and charnockites, as well as similar T(DM) ages between 2.0 and 1.5 Ga are consistent with evolution from a relatively homogeneous and enriched common crustal (metasedimentary) protolith. Results suggest a genetic link between metatexites, diatexites, orthogneisses and charnockites and a two-step process for charnockite development: (a) generation of the hydrated igneous protoliths by anatexis of metasedimentary rocks; (b) continuous high-grade metamorphism that transformed the ""S-type granitoids"" (leucosomes and diatexites) into orthogneisses and, as metamorphism and dehydration progressed, into charnockites. (C) 2011 Elsevier Ltd. All rights reserved.

Metamorphic P-T evolution of granulites in the central Ribeira Fold Belt, SE Brazil

Pseudosections, geothermobarometric estimates and careful petrographic observations of gneissic migmatites and granulites from Neoproterozoic central Ribeira Fold Belt (SE Brazil) were performed in order to quantify the metamorphic P-T conditions during prograde and retrograde evolution of the Brasiliano Orogeny. Results establish a prograde metamorphic trajectory from amphibolite facies conditions to metamorphic peak (T = 850 +/- 50 A degrees C; P = 8 +/- 1 kbar) that promoted widespread dehydrationmelting of 30 to 40% of the gneisses and high-grade granitization. After the metamorphic peak, migmatites evolved with cooling and decompression to T a parts per thousand 500 A degrees C and P a parts per thousand 5 kbar coupled with aH2O increase, replacing the high-grade paragenesis plagioclase-quartz-K-feldspar-garnet by quartz-biotite-sillimanite-(muscovite). Cordierite absence, microtextural observations and P-T results constrain the migmatite metamorphic evolution in the pseudosections as a clockwise P-T path with retrograde cooling and decompression. High-temperature conditions further dehydrated the lower crust with biotite and amphibole-dehydration melting and granulite formation coupled with 10% melt generation. Granulites can thus be envisaged as middle to lower crust dehydrated restites. Granulites were slowly (nearly isobarically) cooled, followed by late exhumation/retrograde rapid decompression and cooling, reflecting a two step P-T path. This retrograde evolution, coupled with water influx, chemically reequilibrated the rocks from granulite to amphibolite/greenschist facies, promoting the replacement of the plagioclase-quartz-garnet-hypersthene peak assemblage by quartz-biotite- K-feldspar symplectites.