Eliminação urinaria do cloreto de sodio na anemia ancilostomotica

Os autores verificaram, em três casos de anemia ancilostomótica, diminuição acentuada na eliminação de cloreto de sódio na urina (2.5 g NaCl em 1.000 cm3 de urina) e progressivo retôrono à normalidade, após administação isolada de sais de ferro, sem eliminação dos helmintos do intestino.

Distribuição geográfica da fauna e flora da Baía de Guanabara

The author studied, the horizontal and vertical distribution of most common part of the flora and fauna of the bay of Guanabara at Rio de Janeiro. In this paper the eulittoral, poly, meso and oligohaline regions were localised and studied; and the first chart of its distribution was presented (fig. 2). The salinity of superficial waters was established through determinations based on 30 trips inside the buy for collecting biological materials. Some often 409 determinations which were previous reported together with the present ones served for the eleboration of a salinity map of the bay of Guanabara (fig. 1). This map of fig. 2 shows the geographic locations of the water regions. EULITTORAL WATER REGIME — Fig. 3 shows the diagram scheme of fauna and flora of this regime. Sea water salinity 34/1.000, density mean 1.027, transparent greenish waters, sea coast with moderate bursting waves. Limpid sea shore with white sand, gneiss with the big barnacle Tetraclita squamosa var. stalactifera (Lam. Pilsbry. Vertical distributions: barna¬cles layers with a green region in which are present the oyster Ostrea pa-rasitica L., the barnacles Tetraclita, Chthamalus, Balanus tintinnabulum var. tintinnabulum (L.) e var. antillensis Pilsbry in connection with several mollusca and the sea beatle Isopoda Lygia sp. Covered by water and exposed to air by the tidal ritms, there is a stratum of brown animals that is the layer of mussels Mytilus perna L., with others brown and chestnut animals : the Crustacea Pachygrapsus, the little crab Porcellana sp., the stone crab Me-nippe nodifrons Stimpson, the sea stars Echinaster brasiliensis (Mull. & Tr.), Astropecten sp. and the sea anemones Actinia sp. Underneath and never visible there is a subtidal region with green tubular algae of genus Codium and amidst its bunches the sea urchin Lycthchinus variegatus (Agass.) walks and more deeply there are numerous sand-dollars Encope emarginata (Leske). The microplancton of this regime is Ceratiumplancton. POLYHALINE WATER REGIMB — Water almost sea water, but directly influenced by continental lands, with rock salts dissolved and in suspension. Salinity: 33 to 32/1.000. This waters endure the actions of the popular nicknamed «water of the hill» (as the waters of mesohaline and oligohaline regimes), becoming suddenly reddish during several hours. That pheno¬menon returns several times in the year and come with great mortality of fishes. In these waters, according to Dr. J. G. FARIA there are species of Protozoa : Peridinea, the Glenoidinium trochoideum St., followed by its satellites which he thinks that they are able to secret toxical substances which can slaughter some species of fishes. In these «waters of the hill» was found a species of Copepoda the Charlesia darwini. In August 1946 the west shore of the Guanabara was plenty of killed fishes occupying a area of 8 feet large by 3 nautical miles of lenght. The enclosure for catching fishes in the rivers mouthes presents in these periods mass dead fishes. The phenomenon of «waters of the hill» appears with the first rains after a period of long dryness. MESOHALINE WATER REGIME — Fig. 4 shows the the diagramm scheme. Salt or brackish water from 30 to 17/1.000 salinity, sometimes until 10/1.000. Turbid waters with mud in suspension, chestnut, claveyous waters; shore dirty black mud without waving bursting; the waters are warmer and shorner than those of the polihaline regime. Mangrove shore with the mangrove trees : Rhizophora mangle L., Avicennia sp., Laguncularia sp., and the »cotton tree of sea» Hibiscus sp. Fauna: the great land crab «guaimú» Cardisoma guanhumi Latr., ashore in dry firm land. There is the real land crab Ucides cordatus (L.) in wetting mud and in neigh¬ bourhood of the burrows of the fiddler-crabs of genus Uca. On stones and in the roots of the Rhizophora inhabits the brightly colored mangrove-tree-crab («aratu» Portuguese nickname) Goniopsis cruentata (Latreille) and the sparingly the big oyster Ostrea rhizophorae Guild. Lower is the region of barnacles Balanus amphitrite var. communis Darwin and var. niveus Darwin; Balanus tintinnabulum var. tintinnabulum (L.) doesn't grow in this brackish water; lower is the region of Pelecipoda with prepollency of Venus and Cytherea shell-fishes and the Panopeus mud crab; there are the sea lettuce Ulva and the Gastreropod Cerithium. The Paguridae Clibanarius which lives in the empty shells of Gasteropod molluscs, and the sessile ascidians Tethium plicatum (Lesuer) appears in some seasons. In the bottom there is a black argillous mud where the «one landed shrimps» Alpheus sp. is hidden. OLIGOHALINE WATER REGIME — The salinity is lower than 10/1.000. average 8/1.000. There are no barnacles and no sea-beetles Isopods of genus Lygia; on the hay of the shore there are several graminea. This brackish water pervades by mouthes of rivers and penetrates until about 3 kilometers river above. While there is some salt dissolved in water, there are some mud crabs of the genus Uca, Sesarma, Metasesarma and Chasmagnatus. The presence of floating green plants coming from the rivers in the waters of a region indicated the oligohaline waters, with low salt content because when the average of NaCl increases above 8/1.000 these plants die and become rusty colored.

Sôbre u'a modificação do meio de Monteverde

It is well known that the culture media used in the presumptive diagnosis of suspiciuous colonies from plates inoculated with stools for isolation of enteric organisms do not always correctly indicate the major groups of enterobacteria. In an effort to obtain a medium affording more exact indications, several media (1-9) have been tested. Modifications of some of these media have also been tested with the result that a satisfactory modification of Monteverde's medium was finaly selected. This proved to be most satisfactory, affording, as a result of only one inoculation, a complete series of basic indications. The modification involves changes in the formula, in the method of preparation and in the manner of storage. The formulae are: A. Thymol blue indicator: NaOH 0.1/N .............. 34.4 ml; Thymol blue .............. 1.6 g; Water .................... 65.6 ml. B. Andrade's indicator. C. Urea and sugar solution: Urea ..................... 20 g; Lactose ................... 30 g; Sucrose ................... 30 g; Water .................... 100 ml. The mixture (C.) should be warmed slightly in order to dissolve the ingredients rapidly. Sterilise by filtration (Seitz). Keep stock in refrigeratior. The modification of Monteverde's medium is prepared in two parts. Semi-solid part - Peptone (Difco) 2.0 g; NaCl 0.5 g; Agar 0.5 g; Water 100.0 ml. Boil to dissolve the ingredients. Adjust pH with NaOH to 7.3-7.4. Boil again for precipitation. Filter through cotton. Ad indicators "A" 0.3 ml and "B" 1.0 ml. Sterilise in autoclave 115ºC, 15 minutes in amounts not higher than 200 ml. Just before using, add solution "C" asseptically in amounts of 10 ml to 200 ml of the melted semi-solid medium, maintained at 48-50ºC. Solid part - Peptone (Difco) 1.5 g; Trypticase (BBL) 0.5 g; Agar 2.0 g; Water 100,00 ml. Boil to dissolve the ingredients. Adjust pH with NaOH to 7.3-7.4. Boils again. Filter through cotton. Add indicators "A" 0.3 ml and "B" 1.0 ml; ferrous ammonium sulfate 0.02 g; sodiun thiosulfate 0.02 g. Sterilise in autoclave 115ºC, 15 minutes in amounts not higher than 200 ml. Just before using, add solution "C" asseptically in amounts of 10 ml to 200 ml of the melted solid medium, maintained at 48-50ºC. Final medium - The semi-solid part is dispensed first (tubes about 12 x 120 mm) in 2.5 ml amounts and left to harden at room temperature, in vertical position. The solid part is dispensed over the hardened semi-solid one in amounts from 2.0 ml to 2.5 ml and left to harden in slant position, affording a butt of 12 to 15 mm. The tubes of medium should be subjected to a sterility test in the incubator, overnight. Tubes showing spontaneous gas bubbles (air) should then be discarded. The medium should be stored in the incubator (37ºC), for not more than 2 to 4 days. Storage of the tubes in the ice-box produces the absorption of air which is released as bubbles when the tubes are incubated at 37ºC after inoculation. This fact confirmed the observation of ARCHAMBAULT & McCRADY (10) who worked with liquid media and the aplication of their observation was found to be essential to the proper working conditions of this double-layer medium. Inoculation - The inoculation is made by means of a long straight needle, as is usually done on the triple sugar, but the needel should penetrate only to about half of the height of the semi-solid column. Indol detection - After inoculation, a strip of sterelized filter papaer previously moistened with Ehrlich's reagent, is suspended above the surface of the medium, being held between the cotton plug and the tube. Indications given - In addition to providing a mass of organisms on the slant for serological invetigations, the medium gives the following indications: 1. Acid from lactose and/or sucrose (red, of yellowsh with strains which reduce the indicators). 2. Gas from lactose and/or sucrose (bubbles). 3. H[2]S production, observed on the solid part (black). 4. Motility observed on the semi-solid part (tubidity). 5. Urease production, observed on solid and semi-solid parts (blue). 6. Indol production, observed on the strip of filter paper (red or purplish). Indol production is not observed with indol positive strains which rapidly acidify the surface o the slant, and the use of oxalic acid has proved to give less sensitive reaction (11). Reading of results - In most cases overnight incubation is enough; sometimes the reactions appear within only a few hours of incubation, affording a definitive orientation of the diagnosis. With some cultures it is necessary to observe the medium during 48 hours of incubation. A description showing typical differential reaction follows: Salmonella: Color of the medium unchanged, with blackening of the solid part when H[2]S is positive. The slant tends to alkalinity (greenish of bluish). Gas always absent. Indol negative. Motility positive or negative. Shigella: Color of the medium unchanged at the beginning of incubation period, but acquiring a red color when the strain is late lactose/sucrose positive. Slant tending to alkalinity (greenish or purplish). Indol positive or negative. Motility, gas and H[2]S always negative. Proteus: Color of the medium generally changes entirely to blue or sometimes to green (urease positive delayed), with blackening of solid part when H[2]S is positive. Motility positive of negative. Indol positive. Gas positive or negative. The strains which attack rapidly sucrose may give a yellow-greenish color to the medium. Sometimes the intense blue color of the medium renders difficult the reading of the H[2]S production. Escherichiae and Klebsiellae: Color of the medium red or yellow (acid) with great and rapid production of gas. Motility positive or negative. Indol generally impossible to observe. Paracoli: Those lactose of sucrose positive give the same reaction as Esherichia. Those lactose or sucrose negatives give the same reactions as Salmonellae. Sometimes indol positive and H[2]S negative. Pseudomonas: Color of the medium unchanged. The slant tends to alkalinity. It is impossible to observe motility because there is no growth in the bottom. Alkaligenes: Color of the medium unchanged. The slant tends to alkalinity. The medium does not alter the antigenic properties of the strains and with the mass of organisms on the slant we can make the serologic diagnosis. It is admitted that this medium is somewhat more laborious to prepare than others used for similar purposes. Nevertheless it can give informations generally obtained by two or three other media. Its use represents much saving in time, labor and material, and we suggest it for routine laboratory work in which a quick presumptive preliminary grouping of enteric organisms is needed.

Estudo microbiológico do sal (Cloreto de sódio) de origem marinha

Foram analisadas 19 amostras de SAL (NaCl), tipo grosso, oriundas de diferentes salinas, para verificação da flora microbiana e presença de bactérias nocivas em Microbiologia Alimentar. O material apresentou grande contaminação por microrganismos saprófitas, bactérias aeróbias e anaeróbias, Gram positivas e negativas, proteolíticas, pigmentadas, esporuladas, leveduras e fungos. A alta incidência das bactérias halofílicas "vermelhas", responsáveis pela deterioração de carnes, pescados e outros produtos salgados foi estudada. A freqüência, em 15 amostras de SAL, de bactérias esporuladas termorresistentes foi calculada em 33%, possuindo um SAL germe termofílico. Para anaeróbios a positividade foi de 80%, havendo esporulação em 40% das culturas isoladas. Os índices para leveduras e fungos foram de 73% e 93%, respectivamente.

Mejora de la eficiencia del uso del agua y los fertilizantes en viveros de plantas ornamentales para jardinería y paisajismo

La producción de plantas ornamentales para jardinería y paisajismo ha experimentado una expansión considerable en todo el territorio español en el último decenio y particularmente en Catalunya. Refiriéndonos como ejemplo al viverismo ornamental de la demarcación de Girona en Catalunya, que el grupo proponente conoce con mayor detalle y se trata de la demarcación peninsular con mayor concentración y producción de leñosas ornamentales y forestales. Cabe destacar que la superficie de producción sobrepasaba las 800 ha a finales de 1999 y que el 90% de los viveristas se agrupaban mayoritariamente en la “Associació de Viveristes de Girona” (AVG), que integraba cerca de 50 asociados con un volumen de producción cercano a los 3000 millones de pesetas y una gama cercana a las 500 especies, presentadas en una gran variedad de formatos. Del conocimiento de este colectivo de viveristas, como resultado de actividades I+D+T previas, se conocía antes de iniciar del proyecto que la práctica de la fertirrigación mediante sistemas de riego de alta frecuencia (goteo o microaspersión) estaba relativamente poco difundida, pero el uso de fertilización mediante abonos de lenta liberación era una práctica común. Algunas evaluaciones de las eficiencias en el uso del agua y de los fertilizantes en dicho colectivo de viveristas indicaban que en general eran bajas y que los equipos y sensores para la automatización del riego eran escasos. Por otra parte los sustratos utilizados eran muy diversos y no se daba una cultura común en cuanto al aprovisionamiento, formulación, enmienda, fertilización de fondo de los sustratos y tampoco en el seguimiento y diagnóstico de la fertilidad de los mismos. Con este proyecto se ha pretendido contribuir a la mejora de la eficiencia del uso del agua y de los nutrientes en los viveros haciendo énfasis especial en el cultivo de especies leñosas autóctonas, empleando sustratos autóctonos y contando como herramientas la fertirrigación, su gestión automática (mediante sensores y actuadores apropiados) y el diagnóstico de la fertilidad del sustrato.

Pharmacokinetic aspects of Tert-Butylaminoethyl disulfide, an experimental drug against schistosomiasis in mice

A preliminary study of the pharmacokinetic parameters of t-Butylaminoethyl disulfide was performed after administration of two different single doses (35 and 300 mg/kg) of either the cold or labelled drug. Plasma or blood samples were treated with dithiothreitol, perchloric acid, and, after filtration, submitted to further purification with anionic resein. In the final step, the drug was retained on a cationic resin column, eluted with NaCl 1M and detected according to the method of Ellman (1958). Alternatively, radioactive drug was detected by liquid scintillation counting. The results corresponding to the smaller dose of total drug suggested a pharmacokinetic behavior related to a one open compartment model with the following parameters: area under the intravenous curve (AUC i.v.):671 ± 14; AUC oral: 150 ± 40 µg.min. ml [raised to the power of -1]; elimination rate constant: 0.071 min [raised to the power of -1]; biological half life: 9.8 min; distribution volume: 0.74 ml/g. For the higher dose, the results seemed to obey a more complex undertermined model. Combining the results, the occurence of a dose-dependent pharmacokinetic behavior is suggested, the drug being rapidly absorbed and rapidly eliminated; the elimination process being related mainly to metabolization. The drug seems to be more toxic when administered I.V. because by this route it escapes first pass metabolism, while being quickly distributed to tissues. The maximum tolerated blood level seems to be around 16 µg/ml.

Estudi de la reactivitat respecte l'oxigen molecular d'una nova família de complexos dinuclears de coure(I) amb lligands hexaazamacrocíclics. Estudis preliminars d'oxidacions de fenols i sulfurs

Projecte de recerca elaborat a partir d’una estada a la Università degli Studi di Pavia entre setembre i desembre del 2007. Els lligands macrocíclics són sistemes que han estat àmpliament utilitzats per mimetitzar els centres actius de proteïnes dinuclears de coure com ara oxigenases (per exemple la tirosinasa). S’ha observat que la reactivitat del complex model depèn de les característiques del lligand com ara la geometria, la cavitat o la distància intermetàl•lica. En aquest sentit, es van sintetitzar dues noves famílies de lligands hexaazamacrocíclics ditòpics, una amb braços laterals tipus piridil i l’altra amb braços laterals tipus fenol. Dintre de cada família, els lligands es diferencien en la cavitat macrocíclica. En aquest treball s’han realitzat estudis de ractivitat respecte de l’oxigen molecular dels complexos dinuclears de coure(I) amb els lligands sintetitzats. Quan es deixen reaccionar els complexos amb oxigen molecular a baixa temperatura, no s’ha observat l’acumulació de cap espècie intermèdia, però si una diferència de reactivitat entre les dues famílies, essent els complexos amb els lligands fenolics molt més reactius que els complexos amb els lligands piridina. També s’han realitzat estudis per comprovar l’activitat oxigenasa dels complexos amb substrats externs. S’han assajat el para-clorofenol i la tioanisole. En el cas del p-clorofenol no s’ha observat en cap cas el producte d’oxigenació corresponent (p-clorocatecol). Quan s’han realitzat els experiments amb la tioanisole, en tots tres casos s’ha observat l’aparició del producte d’oxigenació corresponent (metilfenilsulfòxid), però amb una marcada diferència en el rendiment, mostrant-se molt més actius de nou els complexos amb el lligand fenòlic que el complex amb el lligand piridina.

Preparació de nous sulfurs quirals i aplicació d’aquests en síntesi asimètrica

Estudi realitzat a partir d’una estada al School of Chemistry de la University of Bristol, Gran Bretanya, entre 2006 i 2008. L’epoxidació asimètrica mediada per ilurs de sofre és actualment un mètode ben establert i complementari a les epoxidacions asimètriques d’alquens. Els ilurs de sofre han estat utilitzats també en reaccions d’aziridinació. Aquestes reaccions es poden dur a terme utilitzant tant quantitats subestequiomètriques d’un sulfur en processos catalítics com en quantitats estequiomètriques en forma de sal de sulfoni. Aquest últim procés permet la utilització d’un ventall més ample de substrats. Des de fa anys, s’han desenvolupat una sèrie de sulfurs quirals per aquest tipus de reaccions, alguns dels quals han permès d’obtenir als nivells d’enantioselectivitat. No obstant, la disponibiltat de sulfurs quirals, especialment en les reaccions estequiomètriques, resta com el taló d’Aquiles del procés i ha fet que la seva utilització en síntesi s’hagi quedat relegada tan sols a aquells grups de recerca que han estat pioners en la seva obtenció. Idealment, els sulfurs quirals s’haurien de poder obtenir en poques etapes sintètiques a partir de reactius barats i fàcilment assequibles del « pool quiral » ( les dues formes enantiomèriques accessibles, preferiblement). La seva arquitectura ben definida hauria de permetre d’obtenir alts nivells d’enantioselectivitat. Seguint aquestes premises, durant els dos anys de gaudi de beca s’han sintetitzat dos nous sulfurs quirals i han estat provats en diverses reaccions d’epoxidació i d’aziridinació. Inicialment la recerca es va centar en la síntesi de tiomorfolines, utilitzant limonè com a reactiu. Es va obtenir una tiomorfolina que dóna bons resultats en reaccions d’epoxidació i que es pot recuperar fàcilment, un cop la reacció ha acabat, mitjançant un rentat àcid/base. A més, el material recuperat pot ser reciclat i utilitzat en noves reaccions. A continuació, s’ha sintetitzat un nou sulfur quiral en una sola etapa de reacció a partir de dos dels reactius més barats: limonè i sofre. Aquest nou sulfur ha donat resultats excel•lents tant en termes de rendiments com d’enantio- i de diastereoselectivitat en reaccions d’epoxidació i d’aziridinació. Aquest sulfur quiral es pot recuperar fàcilment mitjançant una destil.lació i pot tornar a ser utilitzat.

Pharmacokinetic profile of tert-butylaminoethanethiol

A preliminary study of the pharmacokinetic parameters of t-Butylaminoethanethiol (TBAESH) was performed after administration of a single dose (35 mg/kg) either orally or intravenously. Plasma or blood samples were treated with dithiothreitol, perchloric acid and, after filtration, submitted to further purification with anionic resin. In the final step the drug was retained on a cationic resin column, eluted with NaCl lM and detected according to the method of Ellman (1958). The results suggested a pharmacokinetic behavior related to a one open compartment model with the following values for the total drug: area under the intravenous curve (AUC i.v.): 443(+ ou -) 24.0; AUC oral: 85.5(+ ou -) 14.5 ug min.ml(elevado a -1); elimination rate constant: 0.069(+ ou -) 0.0055 min(elevado a -1), biological half-life: 10.0(+ ou -) 0.80 min; distribution volume 1.15(+ ou -) 0.15 ml/g; biodisponibility: 0.19(+ ou -) 0.02. From a pharmacokinetic standpoint, TBAESH seems to have no advantage over the analogous disulfide compound.

Permian-polysulphide-siderite-barite-haematite deposit Rude in Samoborska Gora Mts., Zagorje-Mid-Transdanubian zone of the Inner Dinarides

Samoborska Gora Mts. is situated within the westernmost part of the Zagorje-Mid-Transdanubian zone of the Internal Dinarides. The Samoborska Gora Mts. predominantly consists of Permian unmetamorphosed siliciclastic sediments and evaporites, overlain by Lower Triassic sediments. Rude mineralisation is hosted by Permian siliciclastic sediments, below gypsum and anhydrite strata. The central part of the deposit consists of a 1.5 km long stratabound mineralisation, grading laterally into ferruginous sandstone and protruding vertically into a gypsum-anhydrite layer. Siderite-polysulphide-barite-quartz veins are located below the stratabound mineralisation. The stratiform part of the deposit is situated above the stratabound and consists of haematite layer with barite concretions and veinlets. Late stage galena-barite veins overprint earlier types of mineralisation. The Rude ore deposit was generated by predominantly NaCl +/- CaCl(2)-H(2)O solutions. Detrital quartz from stratiform mineralisation contains fluid inclusions with salinities between 7 and 11 wt. % NaCl equ., homogenizing between 150 degrees C to 230 degrees C. Stratabound/siderite-polysulphide-barite-quartz vein type mineralisation was derived from solutions with salinities between 5 and 19 wt. % NaCl equ., homogenizing between 60 degrees C and 160 degrees C, while late stage galenabarite veins were precipitated from solutions with salinities between 11 and 16 wt. % NaCl equ., homogenizing between 100 degrees C to 140 degrees C. Fluid inclusion bulk leachate chemistry recorded Na(+)> Mg(2+)>K(+)>Ca(2+)>Li(+) and Cl-> SO(4)(2-) ions. Sulphur isotope composition of barites and overlying gypsum stems from Permian seawater sulphate, supported by increased Br(-) content, which follows successively the seawater evaporation line. The sulphur isotopic composition of sulphides varies between -0.2 and + 12.5 parts per thousand , as a result of thermal reduction of Permian marine sulphate. Ore-forming fluids were produced by hydrothermal convective cells (reflux brine model), and were derived primarily from Permian seawater, modified by evaporation and interaction with Permian sedimentary rocks. Rude deposits in Samoborska Gora Mts. may be declared as a prototype of the Permian siderite-polysulphide-barite deposits (products of rifting along the passive Gondwana margin), in the Inner Dinarides, and their equivalents extending northeastward into the Zagorje-Mid-Transdanubian Zone and the Gemerides, and southeastward to the Hellenide-Albanides.

Marine biotoxins in the Catalan littoral: could biosensors be integrated into monitoring programmes?

Aquest article descriu els sensors enzimàtics i immunosensors electroquímics que s’han desenvolupat als nostres grups per a la detecció de la biotoxina marina àcid okadaic (OA), i discuteix la possibilitat d’integrar-los en programes de seguiment. Els sensors enzimàtics per a OA que es presenten es basen en la inhibició de la proteïna fosfatasa (PP2A) per aquesta toxina i la mesura electroquímica de l’activitat enzimàtica mitjançant l’ús de substrats enzimàtics apropiats, electroquímicament actius després de la seva desfosforació per l’enzim. Els immunosensors electroquímics descrits en aquest article es basen en un enzimoimmunoassaig sobre fase sòlida competitiu indirecte (ciELISA), amb fosfatasa alcalina (ALP) o peroxidasa (HRP) com a marcatges, i un sistema de reciclatge enzimàtic amb diaforasa (DI). Els biosensors presentats aquí s’han aplicat a l’anàlisi de dinoflagel·lats, musclos i ostres. Les validacions preliminars amb assaigs colorimètrics i LC-MS/MS han demostrat la possibilitat d’utilitzar les bioeines desenvolupades per al cribratge preliminar de biotoxines marines en mostres de camp o de cultiu, que ofereixen informació complementària a la cromatografia. En conclusió, tot i que encara cal optimitzar alguns paràmetres experimentals, la integració dels biosensors a programes de seguiment és viable i podria proporcionar avantatges respecte a altres tècniques analítiques pel que fa al temps d’anàlisi, la simplicitat, la selectivitat, la sensibilitat, el fet de poder ser d’un sol ús i l’efectivitat de cost. This article describes the electrochemical enzyme sensors and immunosensors that have been developed by our groups for the detection of marine biotoxin okadaic acid (OA), and discusses the possibility of integrating them into monitoring programmes. The enzyme sensors for OA reported herein are based on the inhibition of immobilised protein phosphatase 2A (PP2A) by this toxin and the electrochemical measurement of the enzyme activity through the use of appropriate enzyme substrates, which are electrochemically active after dephosphorylation by the enzyme. The electrochemical immunosensors described in this article are based on a competitive indirect Enzyme- Linked ImmunoSorbent Assay (ciELISA), using alkaline phosphatase (ALP) or horseradish peroxidase (HRP) as labels, and an enzymatic recycling system with diaphorase (DI). The biosensors presented herein have been applied to the analysis of dinoflagellates, mussels and oysters. Preliminary validations with colorimetric assays and LC-MS/MS have demonstrated the possibility of using the developed biotools for the preliminary screening of marine biotoxins in field or cultured samples, offering complementary information to chromatography. In conclusion, although optimisation of some experimental parameters is still required, the integration of biosensors into monitoring programmes is viable and may provide advantages over other analytical techniques in terms of analysis time, simplicity, selectivity, sensitivity, disposability of electrodes and cost effectiveness.

The role of Malt1 proteolytic activity in T cell activation and lymphoma development

Abstract Long term contact with pathogens induces an adaptive immune response, which is mainly mediated by T and B cells. Antigen-induced activation of T and B cells is an important event, since it facilitates the transition of harmless, low proliferative lymphocytes into powerful and fast expanding cells, which can, if deregulated, be extremely harmful and dangerous for the human body. One of the most important events during lymphocyte activation is the induction of NF-xB activity, a transcription factor that controls not only cytokine secretion, but also lymphocyte proliferation and survival. Recent discoveries identified the CBM complex as the central regulator of NF-xB activity in lymphocytes. The CBM complex consists of the three proteins Carma1, Bcl10 and Malt1, in which Carma1 serves as recruitment platform of the complex and Bcl10 as an adaptor to recruit Malt1 to this platform. But exactly how Malt1 activates NF-x6 is still poorly understood. We discovered that Malt1 is a protease, which cleaves its interaction partner Bcl10 upon T and B cell stimulation. We mapped the Bcl10 cleavage site by single point mutations as well as by a proteomics approach, and used this knowledge to design a fluorogenic Malt1 reporter peptide. With this tool were we able to the first time demonstrate proteolytic activity of Malt1 in vitro, using recombinant Malt1, and in stimulated T cells. Based on similarities to a metacaspase, we designed a Malt1inhibitor, which allowed unto investigate the role of Malt1 activity in T cells. Malt1-inhibited T cells showed a clear defect in NF-xB activity, resulting in impaired IL-2 cytokine secretion levels. We also found a new unexpected role for Bcl10; the blockade of Bcl10 cleavage resulted in a strongly impaired capability of stimulated T cells to adhere to the extracellular matrix protein fibronectin. Because of the central position of the C8M complex, it is not surprising that different lymphomas show abnormal expressions of Carma1, Bcl10 and Malt1. We investigated the role of Malt1 proteolytic activity in the most aggressive subtype of diffuse large B cell lymphomas called ABC, which was described to depend on the expression of Carmal, and frequently carries oncogenic Carmal mutations. We found constitutive high Malt1 activity in all tested ABC cell lines visualized by detection of cleavage products of Malt1 substrates. With the use of the Malt1-inhibitor, we could demonstrate that Malt-inhibition in those cells had two effects. First, the tumor cell proliferation was decreased, most likely because of lower autocrine stimulation by cytokines. Second, we could sensitize the ABC cells towards cell death, which is most likely caused by reduced expression of prosurvival NF-xB target gens. Taken together, we identified Malt1 as a protease in T and B cells, demonstrated its importance for NF-xB signaling and its deregulation in a subtype of diffuse large B cell lymphoma. This could allow the development of a new generation of immunomodulatory and anti-cancer drugs. Résumé Un contact prolongé avec des pathogènes provoque une réponse immunitaire adaptative qui dépend principalement des cellules T et 8. L'activation des lymphocytes T et B, suite à la reconnaissance d'un antigène, est un événement important puisqu'il facilite la transition pour ces cellules d'un état de prolifération limitée et inoffensive à une prolifération soutenue et rapide. Lorsque ce mécanisme est déréglé ìl peut devenir extrêmement nuisible et dangereux pour le corps humain. Un des événement les plus importants lors de l'activation des lymphocytes est l'induction du facteur de transcription NFxB, qui organise la sécrétion de cytokines ainsi que la prolifération et la survie des lymphocytes. Le complexe CBM, composé des trois protéines Carmai, Bc110 et Malt1, a été récemment identifié comme un régulateur central de l'activité de NF-x8 dans les lymphocytes. Carma1 sert de plateforme de recrutement pour ce complexe alors que Bc110 permet d'amener Malt1 dans cette plateforme. Cependant, le rôle exact de Malt1 dans l'activation de NF-tcB reste encore mal compris. Nous avons découvert que Malt1 est une protéase qui clive son partenaire d'interaction BcI10 après stimulation des cellules T et B. Nous avons identifié le site de clivage de BcI10 par une série de mutations ponctuelles ainsi que par une approche protéomique, ce qui nous a permis de fabriquer un peptide reporteur fluorogénique pour mesurer l'activité de Malt1. Grâce à cet outil, nous avons démontré pour la première fois l'activité protéolytique de Malt1 in vitro à l'aide de protéines Malt1 recombinantes ainsi que dans des cellules T stimulées. La ressemblance de Malt1 avec une métacaspase nous a permis de synthétiser un inhibiteur de Malt1 et d'étudier ainsi le rôle de l'activité de Malt1 dans les cellules T. L'inhibition de Malt1 dans les cellules T a révélé un net défaut de l'activité de NF-x8, ayant pour effet une sécrétion réduite de la cytokine IL-2. Nous avons également découvert un rôle inattendu pour Bcl10: en effet, bloquer le clivage de Bcl10 diminue fortement la capacité d'adhésion des cellules T stimulées à la protéine fïbronectine, un composant de la matrice extracellulaire. En raison de la position centrale du complexe CBM, il n'est pas étonnant que le niveau d'expression de Carmai, Bcl10 et Malt1 soit anormal dans plusieurs types de lymphomes. Nous avons examiné le rôle de l'activité protéolytique de Malt1 dans le sous-type le plus agressif des lymphomes B diffus à grandes cellules, appelé sous-type ABC. Ce sous-type de lymphomes dépend de l'expression de Carmai et présente souvent des mutations oncogéniques de Carma1. Nous avons démontré que l'activité de Malt1 était constitutivement élevée dans toutes les lignées cellulaires de type ABC testées, en mettant en évidence la présence de produits de clivage de différents substrats de Malt1. Enfin, l'utilisation de l'inhibiteur de Malt1 nous a permis de démontrer que l'inhibition de Malt1 avait deux effets. Premièrement, une diminution de la prolifération des cellules tumorales, probablement dûe à leur stimulation autocrine par des cytokines fortement réduite. Deuxièmement, une sensibilisation des cellules de type ABC à ia mort cellulaire, vraisemblablement causée par l'expression diminuée de gènes de survie dépendants de NF-tcB. En résumé, nous avons identifié Malt1 comme une protéase dans les cellules T et B, nous avons mis en évidence son importance pour l'activation de NF-xB ainsi que les conséquences du dérèglement de l'activité de Malt1 dans un sous-type de lymphome B diffus à larges cellules. Notre étude ouvre ainsi la voie au développement d'une nouvelle génération de médicaments immunomodulateurs et anti-cancéreux.

Disseny i caracterització d'estructures RF amb printed electronics per a RFID

En els últims anys printed electronics està aixecant un gran interès entre la indústria electrònica. Aquest tipus de procés consisteix en imprimir circuits amb tècniques d'impressió convencionals utilitzant tintes conductores, resistives, dielèctriques o semiconductores sobre substrats flexibles de baix cost com paper o plàstic. Fer servir aquestes tècniques s'espera que suposi una reducció dels costos de producció degut a que és un procés totalment additiu el que fa que sigui més senzill i es redueixi la quantitat de material emprat. El disseny de dispositius bàsics com resistències, condensadors i bobines per posteriorment veure la relació entre simulacions i valors obtinguts ha ocupat la primera part del projecte. La segona s’ha centrat en fer prototips d’antenes per a RFID (Radio Frequency IDentification) amb la tecnologia que es disposa a CEPHIS (Centre de Prototips i Solucions Hardwre-Software). Tot això ha servit per caracteritzar la tecnologia de la que es disposa i saber en quins apartats s’ha de seguir treballant per aconseguir millors prestacions.

Síntesi i caracterització de complexos polipiridílics de ruteni com a catalitzadors de la reacció d'oxidació de l'aigua

Aquest treball constitueix un estudi sobre els complexos polipiridílics de ruteni i les seves aplicacions en catàlisis oxidativa. Concretament es centren en la reacció d’oxidació de l’aigua per produir oxigen molecular (tot i que també poden ser utilitzats de manera eficient sobre altres substrats). Recull, per tant, la síntesi i caracterització d’alguns lligands nitrogenats tridentats de caràcter facial i/o meridional, així com la seva complexació amb Ru.

Avaluació del creixement del fong ligninolític Trametes versicolor sobre diferents suports sòlids per a l'aplicació en tractaments de bioremediació

Els fongs de podridura blanca (WRF - de l'anglès: White-rot fungi) són de gran interès en l'àmbit de la bioremediació per la seva capacitat de degradar la lignina. La lacasa, un dels enzims extracel·lulars que aquests fongs excreten per degradar la lignina, pot ser utilitzada per degradar els contamiants presents en una matriu donada. Trametes versicolor ha estat estudiat per la seva capacitat de produïr aquest enzim en rersidus agrícoles com a substrats. Un primer triatge basat en la producció de CO2, la mesura de l'activitat lacasa i la quantificació de l'ergosterol han permés seleccionar els substrats on es donava un major poder oxidatiu dels cultius inoculats amb T. versicolor. Els posteriors experiments de colonització de sòls, on es monitoritzava l'activitat lacasa i l'ergosterol, han mostrat que T. vesicolor és capaç de colonitzar sòls i que té una major activitat lacasa en condicions no estèrils. També h'ha provat, mitjançant el test ND24, que T. versicolor és capaç de degradar un contaminant emergent, el naproxè, en sòls estèrils i no estèrils esmenats amb residus agrícoles.