Quantify this! Report on a round table discussion on quantitative mass spectrometry in proteomics.

Following the success of the first round table in 2001, the Swiss Proteomic Society has organized two additional specific events during its last two meetings: a proteomic application exercise in 2002 and a round table in 2003. Such events have as their main objective to bring together, around a challenging topic in mass spectrometry, two groups of specialists, those who develop and commercialize mass spectrometry equipment and software, and expert MS users for peptidomics and proteomics studies. The first round table (Geneva, 2001) entitled "Challenges in Mass Spectrometry" was supported by brief oral presentations that stressed critical questions in the field of MS development or applications (Stöcklin and Binz, Proteomics 2002, 2, 825-827). Topics such as (i) direct analysis of complex biological samples, (ii) status and perspectives for MS investigations of noncovalent peptide-ligant interactions; (iii) is it more appropriate to have complementary instruments rather than a universal equipment, (iv) standardization and improvement of the MS signals for protein identification, (v) what would be the new generation of equipment and finally (vi) how to keep hardware and software adapted to MS up-to-date and accessible to all. For the SPS'02 meeting (Lausanne, 2002), a full session alternative event "Proteomic Application Exercise" was proposed. Two different samples were prepared and sent to the different participants: 100 micro g of snake venom (a complex mixture of peptides and proteins) and 10-20 micro g of almost pure recombinant polypeptide derived from the shrimp Penaeus vannamei carrying an heterogeneous post-translational modification (PTM). Among the 15 participants that received the samples blind, eight returned results and most of them were asked to present their results emphasizing the strategy, the manpower and the instrumentation used during the congress (Binz et. al., Proteomics 2003, 3, 1562-1566). It appeared that for the snake venom extract, the quality of the results was not particularly dependant on the strategy used, as all approaches allowed Lication of identification of a certain number of protein families. The genus of the snake was identified in most cases, but the species was ambiguous. Surprisingly, the precise identification of the recombinant almost pure polypeptides appeared to be much more complicated than expected as only one group reported the full sequence. Finally the SPS'03 meeting reported here included a round table on the difficult and challenging task of "Quantification by Mass Spectrometry", a discussion sustained by four selected oral presentations on the use of stable isotopes, electrospray ionization versus matrix-assisted laser desorption/ionization approaches to quantify peptides and proteins in biological fluids, the handling of differential two-dimensional liquid chromatography tandem mass spectrometry data resulting from high throughput experiments, and the quantitative analysis of PTMs. During these three events at the SPS meetings, the impressive quality and quantity of exchanges between the developers and providers of mass spectrometry equipment and software, expert users and the audience, were a key element for the success of these fruitful events and will have definitively paved the way for future round tables and challenging exercises at SPS meetings.

Oxygen consumption of Litopenaeus vannamei juveniles in heterotrophic medium with zero water exchange

This work aimed at determining the dissolved oxygen consumption rate of Litopenaeus vannamei juveniles maintained in a microbial biofloc raceway system at high density with no aeration. Three 4 L bottles were filled for each treatment, sealed hermetically, and placed in an enclosed greenhouse raceway system. Four shrimp (13.2±1.42 g) were assigned to two sets of the bottles, which underwent the following treatments: light conditions with no shrimp; dark conditions with no shrimp; light conditions with shrimp; and dark conditions with shrimp. Dissolved oxygen content was measured every 10 min for 30 min. A quadratic behavior was observed in dissolved oxygen concentration over time. Significant differences for oxigen consumption were observed only at 10 and 20 min between shrimp maintained in the dark and those under light conditions. At 10 min, a higher value was observed in shrimp maintained under light, and at 20 min, in the dark. Significant differences between 10 and 20 min and between 10 and 30 min were observed when oxygen consumption was analyzed over time in the presence of light. Under dark conditions there were significant differences only between 20 and 30 min. Lethal oxygen concentration (0.65 mg L-1) would be reached in less than one hour either under light or dark conditions with no aeration.

Activity patterns and foraging behaviour of Audouin's gulls at the Ebro Delta, NW Mediterranean.

Twenty Audouin´s gulls, Larus audouinii, breeding in the Ebro Delta (NW Mediterranean) were radio-tracked in 1998 to study their foraging behaviour and activity patterns. Some detrimental effects of tagging on the breeding success of the birds were detected, especially when both members of the pair were tagged. The results were actually constrained by the low number of locations due to natural breeding failure and failure in tag emission, as well as the adverse effect of tagging. However, through a combination of aircraft surveys at sea and a fixed station for automatic tracking of the presence of the birds at the colony, novel individual-based information of home ranges and activity patterns was obtained. Trawler fishing activity seemed to influence both the foraging range and habitat use: while trawlers operated, gulls overlapped their fishing grounds with vessels, probably to scavenge on discards. Very few locations were obtained during a trawling moratorium period, although they were all recorded in coastal bays and terrestrial habitats. During the trawling activity period, gulls ranged over a minimum convex polygon area of 2900 km2. Gulls were tracked up to 40 km from the colony, but some individuals were observed beyond 150 km while still breeding. Arrivals and departures from the colony were in accordance with the trawling timetable. However, most birds also showed some nocturnal foraging activity, probably linked to active fishing of clupeoids (following diel migrations) or to the exploitation of purse-seine fishing activity. Foraging trips lasted on average 15 hours: males performed significantly shorter trips than females, which spent more time outside the colony. The proportion of nocturnal time involved in the foraging trips was the same for males and females, but whilst all males initiated their trips both during the day and at night, some females only initiated their trips during the day. Hatching success was found to be related to foraging effort by males. Gulls spent on average ca. 38% of their time budget outside the nesting territory, representing the time devoted mainly to flying, foraging and other activities.

Estudo fitoquímico de Unonopsis lindmanii - Annonaceae, biomonitorado pelo ensaio de toxicidade sobre a Artemia salina leach

Extracts obtained from leaves, seeds and bark of Unonopsis lindmanii were evaluated by means of Brine Shrimp Lethality test (BSL). Through bioassay-guided chromatographic fractionation, liriodenine, an oxoaporphine alkaloid, was isolated from the bark extracts as the bioactive compound. Two additional inactive known alkaloids, unonopsine and lysicamine were also isolated from the bark extracts.

Síntese de 1,3,5-triazinas substituídas e avaliação da toxicidade frente a Artemia Salina leach

The synthesis of ten symmetrically and unsymmetrically substituted 1,3,5-triazines by Phase Transfer Catalysis (PTC) method is described. Their toxicities were determined against Artemia salina Leach. The LD50 values have also been obtained for these compounds.

Estudo fitoquímico das cascas do caule de Duguetia glabriuscula - Annonaceae, biomonitorado pelo ensaio de toxicidade frente a Artemia salina leach

The extract obtained from stem bark of Duguetia glabriuscula - Annonaceae was evaluated by Brine Shrimp Lethality test (BSL). The bioactive compounds, oxobufoline and lanuginosine, two oxoaporphine alkaloids were isolated by activity-guided fractionation. In addition, the compounds asaraldehyde, (+)-allo-aromadendrane-10beta, 14-diol, and two aporphine alkaloids, polyalthine and oliveridine were also obtained.

Flavonóides e triterpenos de Baccharis pseudotenuifolia: bioatividade sobre Artemia salina

The phytochemical investigation of the chloroformic and methanolic extracts of the Baccharis pseudotenuifolia led to the isolation of triterpenes, steroids and flavonoids. From chloroformic extract were isolated oleanolic acid and alpha-spinasterol while from methanolic extract were isolated the flavonoids: hispidulin, naringenin, 3'-methoxy-luteolin, apigenin, kaempferol, eriodyctiol, aromadendrin, quercetin, 3'-methoxy-quercetin, quercetin-3-O-rhamnoside and quercetin-3-O-glucoside. The structure of these compounds were identified by IR, CG/MS, ¹H and 13C NMR spectral analysis and comparison with literature. In addition, the extracts were evaluated by means of Brine Shrimp Lethality test and the highier activity was observed in the chloroformic extract.

Estudio multidisciplinario del ecosistema manglar en la comunidad tradicional de Curral Velho : Análisis de los servicios ecosistémicos producidos por los manglares a partir de la percepción de la comunidad de Curral Velho

Existe un número elevado de sectores de la sociedad que identifican los beneficios ofrecidos por el ecosistema del manglar así como la interacción entre ellos y el bienestar humano. Aún así, los servicios ecosistémicos (ESs) ofrecidos por la naturaleza son poco visibles y reconocidos legislativamente. Se puede observar que el manglar es uno de los ecosistemas más productivos del planeta. Tienen gran importancia tanto económica, social, ambiental como cultural, pero en las últimas décadas se ha hecho más notable una degradación progresiva debida, en gran parte, a la cría de camarón en cautividad, la camaronicultura. Por esta causa, se observa que la legislación y las políticas que pretenden gestionar estos ecosistemas, no actúan en consecuencia al no imponer ciertas medidas necesarias para esta vital protección, ni considera la importancia de los ecosistemas para la vida de las comunidades tradicionales que dependen de sus servicios para sobrevivir. Por ello es necesario producir un conocimiento sobre ESs a partir de la percepción por parte de la comunidad y que la comunidad se implique en la resolución de esta problemática, ya sea en la toma de decisiones o mediante ciertas actividades o acciones que puedan denunciar estas prácticas insostenibles. Frente a este contexto, este trabajo fue realizado a partir de la información obtenida en la comunidad de Curral Velho (Ceará, Brasil), con el objetivo de realizar un estudio sobre los ESs proveídos por los manglares a partir de la percepción de la comunidad para demostrar la importancia que éstos tienen para el bienestar de los habitantes de la comunidad que se benefician. Los resultados fueron alcanzados a partir de una combinación de metodologías sociales que hacen posible la participación por parte de los habitantes: free listing, encuestas de valoración, grupos focales y observación participante. En la aplicación de estos métodos fueron identificados nuevos ESs de carácter psicológico por los pescadores/as de la pesca artesanal. Por lo que se concluye que es importante tener en cuenta la opinión de las comunidades adyacentes para preservar los flujos ecosistémicos que proporciona el manglar.

Estudio multidisciplinario del ecosistema manglar en la comunidad tradicional de Curral Velho : Estudio sobre la pesca artesanal y análisis de sus diferencias con la acuicultura del camarón

Curral Velho es una comunitat tradicional, situada al nord-est de Brasil, que manté una relació directa amb els ecosistemes que la envolten. La comunitat depèn del estat de conservació del medi natura, ja que obtenen diferents serveis ecositemics. En aquest territori es desenvolupen dos tipus d’economies amb unes bases molt diferents. Una economia tradicional, desenvolupada per la pròpia comunitat, que es basa en la propietat col·lectiva del territori i en optimitzar a llarg termini els beneficis que s’obtenen del medi; y un altre com la camaronicultura, la base de la qual es la obtenció de beneficis a curt termini i amb un territori de propietat privada. Aquesta superposició de models de producció genera impactes ambientals, i un conflicte socio-ambiental entre la comunitat i els que desenvolupen la camaronicultura. L’objectiu es realitzar un estudi econòmic de la pesca artesanal de Curral Velho caracteritzant manera de viure, creant una base de dades sobre pesca artesanal i elaborant indicador de beneficis econòmics generats per la pesca artesanal. Per contextualitzar els resultats es va fer un anàlisis de les dos economies existents a la comunitat. Els resultats obtinguts en primer lloc són que l’economia d’explotació intensiva aporta més guanys per les persones de la comunitat que tenen un relació directa que les que es dediquen a la pesca artesanal, però es important no aturar-se aquí: s’ha de realitzar un anàlisis més profund. Com a conclusió, la activitat pesquera es més rentable a llarg termini ja que els recursos extrets de manera sostenible i així són il·limitats y accessibles a tota la comunitat. A diferència de la camaronicultura, la pesca artesanal no genera desigualtats socials ni vulneracions dels drets humans. Tot el contrari, genera forts vincles entre els individus de la comunitat basats en el treball en equip i l’aprenentatge vivencial e intergeneracional.

Constituintes químicos da raiz e do talo da folha do açaí (Euterpe precatoria Mart., Arecaceae)

Phytochemical investigation of the hexane, ethyl acetate and methanolic extracts of roots and leaf stalks of Euterpe precatoria Mart. ("açaí"), afforded stigmast-4-en-6beta-ol-3-one (3); p-hydroxy benzoic acid (4); 3beta-O-D-glucopyranosyl-sitosterol (5); beta-sitosterol palmitate (6); mixtures of beta-sitosterol and stigmasterol (1 and 2), alpha-, beta-amirin and lupeol (7, 8 and 9), friedelin-3-one and 28-hydroxy-friedelin-3-one (10 and 11) and alpha-, beta-D-glucose (12, 13). Except for 1, 2 and 4, the other isolated constituents are described in the genus for the first time. Compounds 3 and 5 gave good results in the brine shrimp bioassay, which detects compounds with potential uses as antitumor agents, pesticides, etc..

Deep-water macroalgal-dominated coastal detritic assemblages on the continental shelf off Mallorca and Menorca (Balearic Islands, Western Mediterranean)

We present a quantitative physiognomic characterization of major macroalgal-dominated assemblages on coastal detritic bottoms of the continental shelf off Mallorca and Menorca (Balearic Islands, Western Mediterranean). In late spring of 2007 and 2008, 29 samples were collected by bottom trawling at depths between -52 and -93 m. These samples were then sorted and identified to their lowest taxonomic level. Statistical analyses distinguished six different assemblage types: shallower water environments (-52 to -65 m in depth) were characterized by Osmundaria volubilis and Phyllophora crispa meadows and two types of Peyssonnelia beds; two assemblage types, Laminaria rodriguezii beds and maërl beds, were only present in deep-water environments (-77 to -81 m); and an assemblage dominated by P. crispa and Halopteris filicina was found in both shallow and deep waters (-57 to -93 m). We assess the distribution of these six assemblage types through the studied area.

Antioxidant and cytotoxic activities of 'açaí' (Euterpe precatoria Mart.)

Decoction of roots of the Amazonian palm açaí (Euterpe precatoria Mart.) is widely used by Brazilian and Peruvian people as an anti-inflammatory, to heal kidney and liver diseases and against snake bites. In this study, the antioxidant activity of extracts and flavonoids (quercetin, catechin, epicatechin, rutin and astilbin) isolated from roots and leaf stalks of E. precatoria was investigated using β-carotene in TLC plates and DPPH radical scavenging in a spectrophotometric bioassay. All extracts and flavonoids showed activity. Also, the cytotoxic activity of these extracts was evaluated by the brine shrimp (Artemia salina) larvicide bioassay and was lower than that of lapachol, used as control. The presence of flavonoids and sitosterol-3-O-β-D-glucopyranoside in the extracts can justify the use of the plant in traditional medicine.

Toxicidade e atividade antioxidante de flavonoides das cascas das raízes de Lonchocarpus filipes

The phytochemical investigation of dichloromethane extract from root bark of Lonchocarpus filipes Benth (Leguminosae) afforded four flavonoids including three dibenzoylmethane derivatives rarely found in nature. The structures were established based on their spectral data (¹H and 13C NMR, 2D-NMR) as being: lanceolatin B (1), pongamol (2), (E)-7-O-methylpongamol (3) and (E)-9-O-methylpongamol (4). Compound (4) is described herein for the first time as a natural product. The extracts and the isolated compounds (1), (2) and (3) displayed high toxicity in the brine shrimp lethality assay. Only compound (2) showed antioxidant activity using a DPPH radical scavenging assay. This is the first report on the phytochemical study of Lonchocarpus filipes.

Purificação e caracterização da quitinase de uva (Vitis vinífera L. cv Red Globe) para a produção de quitosana a partir de quitina de camarão

Chitinase is produced by a wide variety of plants as a defense against peste attacks. In this study, grape chitinases were purified 16 times by fractionation in 80% ammonium sulfate followed by dialysis and filtration. Purified chitinases exhibited enzymatic activity toward chitin azure. The yield of purified chitinase was 229 mg/L with chitinase activity of 563 U/g. Chitinases had molecular masses of 24 and 30 kDa, as evaluated by SDS-PAGE 12.5%. Two pH optima were determined 3.0 and 6.0. The optimal temperature was 42 °C. Pre hydrolysis of crystalline shrimp chitin by chitinases caused in an increase in the deacetylation ratio triggered by chitin deacetylase producing chitooligosaccharides with DA (degree acetylation) of 58.8%.

Otimização e validação de método multirresíduo para determinação de sulfonamidas em camarão cultivado por cromatografia líquida de alta eficiência com detecção por UV

This work reports the optimization and method validation for sulfonamides (sulfamethazine, sulfaquinoxaline, sulfadimethoxine and sulfathiazole) in shrimp muscle using HPLC-UV. The sulfonamides were extracted with acetonitrile and acetic acid, and the extract cleaned up with a Strata SCX SPE cartridge prior to analysis. The method presented linearity in the range of 20-120 µg kg-1, good linear correlation (r > 0.99), and limits of quantification in the range of 4.7-20.2 µg kg-1. The recovery for shrimp muscles spiked with 50-150 µg kg-1 ranged from 63.2-108.0%. Precision and accuracy analysis showed acceptable relative standard deviation. Commercial shrimps were analyzed and sulfonamides don't were found above of the method limit of quantification.