Particle flux measured on deep sea sediment trap VP-2_trap (Appendix A2.7)

A 17 month record of vertical particle flux of dry weight, carbonate and organic carbon were 25.8, 9.4 and 2.4g/m**2/y, respectively. Parallel to trap deployments, pelagic system structure was recorded with high vertical and temporal resolution. Within a distinct seasonal cycle of vertical particle flux, zooplankton faecal pellets of various sizes, shapes and contents were collected by the traps in different proportions and quantities throughout the year (range: 0-4,500 10**3/m**2/d). The remains of different groups of organisms showed distinct seasonal variations in abundance. In early summer there was a small maximum in the diatom flux and this was followed by pulses of tinntinids, radiolarians, foraminiferans and pteropods between July and November. Food web interactions in the water column were important in controlling the quality and quantity of sinking materials. For example, changes in the population structure of dominant herbivores, the break-down of regenerating summer populations of microflagellates and protozooplankton and the collapse of a pteropod dominated community, each resulted in marked sedimentation pulses. These data from the Norwegian Sea indicate those mechanisms which either accelerate or counteract loss of material via sedimentation. These involve variations in the structure of the pelagic system and they operatè on long (e.g. annual plankton succession) and short (e.g. the end of new production, sporadic grazing of swarm feeders) time scales. Connecting investigation of the water column with a high resolution in time in parallel with drifting sediment trap deployments and shipboard experiments with the dominant zooplankters is a promising approach for giving a better understanding of both the origin and the fate of material sinking to the sea floor.

Trace element composition of IODP Site 304-U1309 and 305-U1309 (Table A2)

IODP Site U1309 was drilled at Atlantis Massif, an oceanic core complex, at 30°N on the Mid-Atlantic Ridge (MAR). We present the results of a bulk rock geochemical study (major and trace elements) carried out on 228 samples representative of the different lithologies sampled at this location. Over 96% of Hole U1309D is made up of gabbroic rocks. Diabases and basalts cross-cut the upper part of the section; they have depleted MORB compositions similar to basalts sampled at MAR 30°N. Relics of mantle were recovered at shallow depth. Mantle peridotites show petrographic and geochemical evidence of extensive melt-rock interactions. Gabbroic rocks comprise: olivine-rich troctolites (> 70% modal olivine) and troctolites having high Mg# (82-89), high Ni (up to 2300 ppm) and depleted trace element compositions (Yb 0.06-0.8 ppm); olivine gabbros and gabbros (including gabbronorites) with Mg# of 60-86 and low trace element contents (Yb 0.125-2.5 ppm); and oxide gabbros and leucocratic dykes with low Mg# (< 50), low Ni (~65 ppm) and high trace element contents (Yb up to 26 ppm). Troctolites and gabbros are amongst the most primitive and depleted oceanic gabbroic rocks. The main geochemical characteristics of Site U1309 gabbroic rocks are consistent with a formation as a cumulate sequence after a common parental MORB melt, although (lack of systematic) downhole variations indicate that the gabbroic series were built by multiple magma injections. In detail, textural and geochemical variations in olivine-rich troctolites and gabbronorites suggest chemical interaction (assimilation?) between the parental melt and the intruded lithosphere. Site U1309 gabbroic rocks do not represent the complementary magmatic product of 30°N volcanics, although they sample the same mantle source. The bulk trace element composition of Site U1309 gabbroic rocks is similar to primitive MORB melt compositions; this implies that there was no large scale removal of melts from this gabbro section. The occurrence of such a large magmatic sequence implies that a high magmatic activity is associated with the formation of Atlantis Massif. Our results suggest that almost all melts feeding this magmatic system stays trapped into the intruded lithosphere.

Molecular Characterization of Three Novel Phospholipase A2 Proteins from the Venom of Atheris chlorechis, Atheris nitschei and Atheris squamigera

Secretory phospholipase A2 (sPLA2) is known as a major component of snake venoms and displays higher-order catalytic hydrolysis functions as well as a wide range of pathological effects. Atheris is not a notoriously dangerous genus of snakes although there are some reports of fatal cases after envenomation due to the effects of coagulation disturbances and hemorrhaging. Molecular characterization of Atheris venom enzymes is incomplete and there are only a few reports in the literature. Here, we report, for the first time, the cloning and characterization of three novel cDNAs encoding phospholipase A2 precursors (one each) from the venoms of the Western bush viper (Atheris chlorechis), the Great Lakes bush viper (Atheris nitschei) and the Variable bush viper (Atheris squamigera), using a “shotgun cloning” strategy. Open-reading frames of respective cloned cDNAs contained putative 16 residue signal peptides and mature proteins composed of 121 to 123 amino acid residues. Alignment of mature protein sequences revealed high degrees of structural conservation and identity with Group II venom PLA2 proteins from other taxa within the Viperidae. Reverse-phase High Performance Liquid Chromatography (HPLC) profiles of these three snake venoms were obtained separately and chromatographic fractions were assessed for phospholipase activity using an egg yolk suspension assay. The molecular masses of mature proteins were all identified as approximately 14 kDa. Mass spectrometric analyses of the fractionated oligopeptides arising from tryptic digestion of intact venom proteins, was performed for further structural characterization.

Lipoprotein-associated phospholipase A2 (Lp-PLA2) as a therapeutic target to prevent retinal vasopermeability during diabetes

Lipoprotein-associated phospholipase A2 (Lp-PLA2) hydrolyses oxidized low-density lipoproteins into proinflammatory products, which can have detrimental effects on vascular function. As a specific inhibitor of Lp-PLA2, darapladib has been shown to be protective against atherogenesis and vascular leakage in diabetic and hypercholesterolemic animal models. This study has investigated whether Lp-PLA2 and its major enzymatic product, lysophosphatidylcholine (LPC), are involved in blood-retinal barrier (BRB) damage during diabetic retinopathy. We assessed BRB protection in diabetic rats through use of species-specific analogs of darapladib. Systemic Lp-PLA2 inhibition using SB-435495 at 10 mg/kg (i.p.) effectively suppressed BRB breakdown in streptozotocin-diabetic Brown Norway rats. This inhibitory effect was comparable to intravitreal VEGF neutralization, and the protection against BRB dysfunction was additive when both targets were inhibited simultaneously. Mechanistic studies in primary brain and retinal microvascular endothelial cells, as well as occluded rat pial microvessels, showed that luminal but not abluminal LPC potently induced permeability, and that this required signaling by the VEGF receptor 2 (VEGFR2). Taken together, this study demonstrates that Lp-PLA2 inhibition can effectively prevent diabetes-mediated BRB dysfunction and that LPC impacts on the retinal vascular endothelium to induce vasopermeability via VEGFR2. Thus, Lp-PLA2 may be a useful therapeutic target for patients with diabetic macular edema (DME), perhaps in combination with currently administered anti-VEGF agents.

Konnektorer i inlärarsvenska på CEFR-nivåerna A1, A2 och B1 – en longitudinell analys ur ett funktionellt perspektiv

Syftet med avhandlingen har varit att granska finska inlärares konnektorbruk på CEFR-nivåerna A1, A2 och B1 longitudinellt ur ett funktionellt perspektiv. Jag har studerat vad som är kännetecknande för konnektorbruket på dessa CEFR-nivåer och i vilken funktion konnektorerna har använts på dessa nivåer. Vidare har jämförts konnektorbruket i materialet med det som sägs i CEFR-kriterierna. Slutligen har jag också granskat hur konnektorbruket utvecklas. Som material har jag använt berättande texter (n=303) skrivna av 101 finskspråkiga grundskolelever och gymnasister. Materialet ingår i projektet Topling – Inlärningsgångar i andraspråket vid Jyväskylä universitet. I avhandlingen har använts såväl kvantitativa som kvalitativa metoder. Jag har räknat konnektorernas och konnektorkategoriernas frekvenser samt analyserat i vilka funktioner konnektorerna har använts. I den funktionella analysen har använts systemisk-funktionell lingvistik (Halliday & Matthiessen 2004) samt Labovs (1972) modell om berättelsestrukturen. Analysen har visat att konnektorbruket skiljer sig mellan CEFR-nivåerna A1, A2 och B1. Antalet konnektorer ökar såväl från nivå A1 till A2 som från nivå A2 till nivå B1 och andelen additiva och målspråksavvikande konnektorer minskar medan andelen temporala, kausala och komparativa konnektorer samt att ökar. Konnektorerna har använts först och främst i deras prototypiska funktioner på alla dessa CEFR-nivåer. Vissa konnektorer (när, eftersom, att) verkar även ha en funktion i berättelsestrukturen. Om man jämför konnektorbruket med CEFR-kriterierna kan man konstatera att inlärare på nivå A1 använder pronomenet den i stället för sedan även om denna konnektor nämns i CEFR-kriterierna på nivå A1. Konnektorbruket verkar utvecklas på det sättet att antalet konnektorer samt andelen additiva, temporala och komparativa konnektorer samt att ökar. Andelen additiva konnektorer och målspråksavvikande konnektorer minskar. Vidare börjar inlärare använda mera olika konnektorer och på nivå B1 även mindre frekventa konnektorer som om och fast. I fortsättningen borde man granska konnektorbruket i olika texttyper samt studera om explicit undervisning påverkar inlärares konnektorbruk.

Caraterização molecular de anemias hipocrómicas e microcíticas com HB A2 normal

Introdução - As anemias hipocrómicas e microcíticas, não sideropénicas, são, na sua maioria, de origem talassémica. As talassémias são hemoglobinopatias caraterizadas pela redução ou ausência da produção de cadeias globínicas, provocadas por mutações nos genes globínicos. A β-talassémia está mais frequentemente associada a mutações pontuais e pequenas deleções ou inserções no gene HBB, enquanto que a α-talassémia normalmente resulta de deleções que eliminam os genes HBA2 e/ou HBA1. Deleções de um só gene, -α3.7 e -α4.2, são frequentes entre africanos, mediterrânicos e asiáticos enquanto que grandes deleções que removem ambos os genes alfa, como a deleção do Sudeste Asiático (--SEA), são comuns nas populações asiáticas. Grandes deleções nos clusters alfa e beta são condições raras, geralmente associadas a fenótipos severos. A identificação destas deleções é realizada através da técnica de MLPA (multiplex ligation-dependent probe amplification). Objetivos - O objetivo principal deste trabalho foi estudar um grupo de indivíduos com hipocromia e microcitose e Hb A2 normal, com suspeita de possuirem deleções nos clusters α ou β. Pretendeu-se, ainda, caraterizar a extensão das deleções encontradas e, quando possível, determinar a localização dos seus breakpoints. Materiais e Métodos - Cinquenta e oito indivíduos (28 homens e 30 mulheres), com hipocromia e microcitose de origem desconhecida, seguidos na Consulta de Hematologia do Hospital Pediátrico e do Hospital Geral do CHC, ou enviados de outros centros, foram testados para a presença de deleções nos clusters α e β. Foram efetuados hemogramas a todas as amostras e os estudos de hemoglobina foram realizados por cromatografia líquida de alta performance (HPLC). Os estudos moleculares incluiram GAP-PCR, MLPA, sequenciação genética e PCR / hibridização reversa. Resultados - Foi possível obter o padrão do rácio das sondas MLPA para as deleções α e β conhecidas ou já caraterizadas. Foram encontradas 6 deleções HBA desconhecidas que removem os genes α ou as regiões reguladoras desse cluster. Detetou-se, também, uma deleção no cluster β, que elimina praticamente todos os seus genes. Conclusões - A identificação de grandes delções nos clusters α e β-globínicos em indivíduos com hipocromia e microcitose, com HbA2 normal, é crucial para o aconselhamento genético. A metodologia de MLPA é uma abordagem simples e fiável, muito útil para o diagnóstico de casos de talassemia, em que não são detetadas mutações α ou β através das técnicas convencionais

Effect of umbelliferone (7-hydroxycoumarin, 7-HOC) on the enzymatic, edematogenic and necrotic activities of secretory phospholipase A2 (sPLA2) isolated from Crotalus durissus collilineatus venom

Flavonoids, coumarins and other polyphenolic compounds are powerful antioxiants both in hydrophilic and lipophylic environments with diverse pharmacological properties including anti-inflammatory activity. Despite being widely used as powerful therapeutic agents for blood coagulation disorders, more specifically to control some serine protease enzymes, the mechanism of anti-inflammatory activity of coumarins is unknown, unlike that of flavonoids. Although their controlling effect on serine proteases is well acknowledged, their action on secretory phospholipase A2 (sPLA2) remains obscure. The present study describes the interaction between umbelliferone (7-HOC) and the sPLA2 from Crotalus durissus collilineatus venom. In vitro inhibition of sPLA2 enzymatic activity by 7-HOC was estimated using 4N3OBA as substrate, resulting in an irreversible decrease in such activity proportional to 7-HOC concentration. The biophysical interaction between 7-HOC and sPLA2 was examined by fluorescent spectral analysis and circular dichroism studies. Results from both techniques clearly showed that 7-HOC strongly modified the secondary structure of this enzyme and CD spectra revealed that it strongly decreased sPLA2 alphahelical conformation. In addition, two-dimensional electrophoresis indicated an evident difference between HPLC-purified native and 7-HOC-treated sPLA2s, which were used in pharmacological experiments to compare their biological activities. In vivo anti-inflammatory activity was assessed by the sPLA2-induced mouse paw edema model, in which 7-HOC presented an effect similar to those of dexamethasone and cyproheptacline against the pro-inflammatory effect induced by native sPLA2 on the mouse paw edema, mast cell degranulation and skin edema. on the other hand, 7-HOC exhibited a more potent inhibitory effect on sPUL2 than that of p-bromophenacyl bromide (p-BPB). Our data suggest that 7-HOC interacts with sPLA2 and causes some structural modifications that lead to a sharp decrease or inhibition of the edematogenic and myotoxic activities of this enzyme, indicating its potential use to suppress inflammation induced by sPLA2 from the snake venom. (C) 2008 Published by Elsevier Ltd.

The effect of phospholipase A2 from Crotalus durissus collilineatus on Leishmania (Leishmania) amazonensis infection

In this study, the effect of phospholipase A2 (PLA2) derived from Crotalus durissus collilineatus was evaluated in vitro and in vivo on experimental cutaneous leishmaniasis. The promastigote and amastigote forms treated with PLA2 presented increased growth rate. In vivo studies showed that PLA2-treated Leishmania (Leishmania) amazonensis promastigotes increased the size of lesions in BALB/c mice, and histopathological analysis showed numerous necrotic regions presenting a higher density of polymorphonuclear, mononuclear, and amastigote cells. Additionally, infected macrophages treated with PLA2 were able to generate prostaglandin E2 (PGE2). Cytokine quantification showed that the supernatant from infected macrophages presented moderate and high amounts of IL-2 and IL-10, respectively. However, in PLA2-treated infected macrophages, suppression of IL-2 levels occurred, but not of IL-10 levels. Observation also revealed that both the supernatant and lysate of L. (L.) amazonensis promastigotes exhibited PLA2 activity, which, in the presence of dexamethasone, showed no reduction in their activities; while glucocorticoid maintained the ability of promastigote forms to infect macrophages, which presented values similar to controls. In conclusion, the results indicate that PLA2 may be a progression factor for cutaneous leishmaniasis, since the PLA2 effect suppressed IL-2 levels and generated PGE2, an inflammatory lipid mediator.

Estudo do papel desempenhado pelo heterotetrâmero de anexina A2 em células de cancro da mama hipóxicas

Dissertação de Mestrado, Biologia Molecular e Microbiana, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2014