La susceptibilitat individual a l’autoimmunitat pot estar determinada per una combinació de polimorfismes específics de gens que codifiquen per a múltiples citocines, per al complex principal d'histocompatibilitat (MHC) , molècules d'adhesió i proteïnes cel·lulars. En aquest treball ens hem volgut centrar en l’associació d’al·lels del MHC amb les malalties immunitàries. L'explicació actual per a aquest tipus d'associacions proposa que les molècules del MHC que s'han trobat associades, uneixen de manera eficient als autoantígens lligats a la malaltia, la qual cosa dóna lloc a una resposta immune perifèrica contra els autoantígens. En aquest treball hem volgut aprofundir en la caracterització dels repertoris de pèptids presentats per molècules de classe II associades a autoimmunitat en humans i en ratolí, a partir dels pèptids naturals associats. Els mètodes de predicció de la unió d’un pèptid a un determinat MHC són importants per poder identificar aquells segments d’una proteïna que seran presentats a les cèl·lules T, i per tant ens poden ser útils a l’hora de generar vacunes peptídiques o pel desenvolupament de teràpies antigen-específiques. En humans, l’al·lel HLA-DR3 és conegut per la seva associació a malalties com la diabetis tipus 1 o la malaltia de Graves-Basedaw (GB) i en ratolíns el complexe H-2d s’associa a malalties tipus th2, entre elles també la malaltia GB, i diversos càncers. En aquest estudi s’ha fet servir el mètode establert al nostre laboratori basat en l’aïllament dels pèptids per immunoafinitat i l’anàlisi per espectròmetria de masses d’alta resolució. / SUMMARY: Individual susceptibility to autoimmunity may be determined by a combination of specific polymorphisms of genes encoding multiple cytokines by the major histocompatibility complex (MHC) proteins and cell adhesion molecules. In this study we wanted to focus the association of MHC alleles with immune diseases. The current explanation for these associations suggests that the MHC molecules that have been associated autoantigens bind efficiently linked to the disease, which leads to a peripheral immune response against those autoantigens. In this study, we wanted to characterize the repertoire of peptides presented by class II molecules associated with autoimmunity in humans and mice from natural peptides. The methods for predicting peptide binding to a particular MHC is important to identify those segments of a protein that will be presented to T cells, and therefore it can be useful in generating peptide vaccines or the development of antigen-specific therapies. In humans, the allele HLA-DR3 is known for its association with type 1 diabetes and Graves disease (GD) and the complex H-2d mice is associated with Th2 type disease as GD and various cancers. For this characterization, we used a method established in our laboratory based on the isolation of peptides by immunoafinity and analysis by high resolution mass spectrometry

Dinoflagellates are one of the most abundant and diverse groups of microalgae. Many dinoflagellates are covered with cellulose plates, whereas others lack these plates and are hence referred to as athecate or unarmoured. Unarmoured species have been historically poorly characterized because they deform when fixed with traditional methods. Most unarmoured genera are included within the order Gymnodiniales and are differentiated by morphological characters, but the recent combination of morphological observations with phylogenetic data concludes that molecular phylogenies do not support the "classical" morphological criteria used to distinguish the genera. It is also known that the order Gymnodiniales is not monophyletic. Consequently, the taxonomy of dinoflagellates has shifted to a combination of morphology and molecular information. The widespread use of molecular techniques has enabled detailed studies on the systematics of a lot of groups. Although many dinoflagellates are not easily cultivable and cannot be sequenced by standard techniques, the single-cell PCR technique has allowed obtaining sequences from a single cell by taking advantage of the large copy number of the ribosomal genes. Extensive identifications of dinoflagellates present in the NW Mediterranean Sea were carried out between the 1960s and 1980s. However, the diversity of unarmoured dinoflagellates was not suitably characterized because of the use of fixed samples and the lack of molecular tools. This thesis studies the diversity of dinoflagellates belonging to the Gymnodiniales order from the Catalan coast, as a representative site of the NW Mediterranean Sea. This thesis presents a revision of their taxonomy by combining morphological studies of live specimens with the respective phylogenetic information. Given that the phylogeny of most of the studied organisms had not been previously determined and the evidence that Gymnodiniales is not monophyletic, a secondary objective was to study the phylogenetic relationships of species. The D1-D2 region of the LSU rDNA sequence was selected to conduct the SC-PCR, but SSU rDNA sequences were obtained when necessary. The combination of morphological and molecular data has led to the identification of 58 unarmoured species belonging to the order Gymnodiniales. Ten morphospecies are detected for the first time in the Mediterranean Sea, and eight, for the first time along the Catalan coast (Chapter 1 and 2). Additionally, the application of single-cell PCR has resulted in the sequencing of 43 unarmoured species, 25 of them for the first time (Chapters 1 and 2). It has also allowed the detection and characterization of species previously undescribed, resulting in the erection of two new species: Gymnodinium litoralis (Chapter 3) and Polykrikos tanit (Chapter 4). Additionally, the toxic species Cochlodinium polykrikoides was detected for the first time along the Catalan coast. Most populations formed a newly differentiated ribotype, but others were included within a previously known one, demonstrating their coexistence in the Mediterranean Sea (Chapter 5). Finally, a Ceratoperidinium specimen was sequenced for the first time and a new phylogenetic clade was obtained, along with other unarmoured dinoflagellates, including Ceratoperidinium margalefii, Gyrodinium falcatum, which was transferred to the genus Ceratoperidinium, three Cochlodinium species, and two Gymnodinium-like organisms. This resulted in the emendation of the Ceratoperidiniaceae family and the genus Ceratoperidinum (Chapter 6). The correct identification of the species has allowed to conclude that there is a high diversity of unarmoured dinoflagellates in the Catalan coast, and to discuss the implications on the distribution and biogeography of the species at a Mediterranean level, as C. polykrikoides (Chapter 5), or globally, as for Gyrodinium spirale (Chapter 2) / Les dinoflagel•lades són un dels grups més abundants i diversos de microalgues. Moltes espècies estan cobertes amb plaques de cel•lulosa, mentre que d’altres que no en tenen són conegudes com a atecades o nues. Les espècies atecades han estat generalment mal caracteritzades ja que es deformen quan són fixades amb mètodes tradicionals. La majoria de gèneres atecats s'inclouen dins l’ordre Gymnodinials i es distingeixen per caràcters morfològics, però la combinació recent d'observacions morfològiques amb dades filogenètiques conclou que les filogènies moleculars no suporten els criteris morfològics clàssics utilitzats per distingir els gèneres. També es coneix que l’ordre Gymnodinials no és monofilètic. Per tant, la correcta taxonomia de dinoflagel•lades requereix una combinació d’informació morfològica i molecular. L'ús generalitzat de tècniques moleculars ha permès estudis detallats sobre la sistemàtica de molts grups. Tot i que moltes dinoflagel•lades no són fàcilment cultivables i no es poden seqüenciar mitjançant tècniques estàndards, la tècnica de single-cell PCR ha permès obtenir seqüències a partir d'una sola cèl•lula, aprofitant el gran nombre de còpies dels gens ribosòmics que les dinoflagel•lades contenen. Entre els anys 1960 i 1980 es van dur a terme identificacions extensives de les dinoflagel•lades existents al litoral català. Tanmateix, no es va caracteritzar adequadament la diversitat de dinoflagel•lades atecades degut a l’ús de mostres fixades i la manca d'eines moleculars. Aquesta memòria presenta l'estudi de la diversitat d'espècies de l’ordre Gymnodinials a la costa catalana com a representativa del NO del Mar Mediterrani, entre els anys 2010-2013. L'estudi presenta una revisió de la seva taxonomia pel que s'han combinant estudis morfològics d’exemplars vius amb la corresponent informació filogenètica. Atès que la filogènia d’alguns dels organismes estudiats no s'havia determinat amb anterioritat i l'evidència que l’ordre Gymnodinials no és monofilètic, el segon objectiu ha estat estudiar les relacions filogenètiques de les espècies. Es va seleccionar la regió D1-D2 del 28s rDNA per dur a terme els anàlisis amb SC-PCR, però també es van obtenir seqüències del 18s rDNA quan ha estat necessari. La combinació de les dades morfològiques i moleculars ha permès la identificació inequívoca de 58 espècies atecades pertanyents a l’ordre Gymnodinials. D’aquestes, es detecten deu morfoespècies per primera vegada a la mar Mediterrània, i vuit per primera vegada al litoral català (Capítols 1 i 2). A més, l'aplicació de la SC-PCR ha permès seqüenciar 43 espècies atecades, 25 de les quals per primera vegada (Capítols 1 i 2). També ha permès la detecció i caracterització d’espècies no descrites prèviament, que ha resultat en la descripció de les noves espècies Gymnodinium litoralis (Capítol 3) i Polykrikos tanit (Capítol 4). A més, es va detectar per primera vegada l'espècie tòxica Cochlodinium polykrikoides al litoral català. La majoria d’aquests organismes pertanyien a un nou ribotip, però d’altres quedaren inclosos en un ribotip ja conegut, demostrant la seva coexistència al Mar Mediterrani (Capítol 5). Finalment, es va seqüenciar per primera vegada un espècimen del gènere Ceratoperidinium i es va obtenir un nou grup filogenètic, juntament amb d’altres dinoflagel•lades atecades, incloent Ceratoperidinium margalefii, Gyrodinium falcatum, que va ser transferit al gènere Ceratoperidinium, tres espècies de Cochlodinium, i dos organismes semblants a Gymnodinium. Això va donar lloc a l'esmena de la família Ceratoperidiniaceae i del gènere Ceratoperidinum (Capítol 6). La correcta identificació de les espècies ha permès concloure que la costa catalana presenta una gran diversitat de dinoflagel•lades atecades, i discutir les implicacions en la distribució i biogeografia de les espècies a nivell de la Mediterrània, com el cas de C. polykrikoides (Capítol 5), o global, com el cas de Gyrodinium spirale (Capítol 2)

The contribution of Clostridium difficile toxin A and B (TcdA and TcdB) to cellular intoxication has been extensively studied, but their impact on bacterial colonization remains unclear. By setting-up two- and three-dimensional in vitro models of polarized gut epithelium, we investigated how C. difficile infection is affected by host cell polarity and whether TcdA and TcdB contribute to such events. Indeed, we observed that C. difficile adhesion and penetration of the epithelial barrier is substantially enhanced in poorly polarized or EGTA-treated cells, indicating that bacteria bind preferentially to the basolateral cell surface. In this context, we demonstrated that sub-lethal concentrations of C. difficile TcdA are able to alter cell polarity by causing redistribution of plasma membrane components between distinct surface domains. Taken together, the data suggest that toxin-mediated modulation of host cell organization may account for the capacity of this opportunistic pathogen to gain access to basolateral receptors leading to a successful colonization of the colonic mucosa.

La calidad de los oocitos es sinónimo de competencia oocitaria y se define como la capacidad de un oocito para reanudar la meiosis, dividirse después de la fertilización, desarrollarse hasta la etapa de blastocisto, inducir la preñez y conseguir el nacimiento de un animal sano. La selección de los oocitos más competentes es un punto clave en los programas de producción in vitro de embriones (PIVE). El test de Azul de Cresol Brillante (BCB) es un método no invasivo que separa los oocitos que han finalizado su crecimiento (BCB+: citoplasma azul) de los oocitos con la enzima G6PDH activa o oocitos en proceso de crecimiento (BCB-: citoplasma incoloro). En esta tesis, hemos llevado a cabo 3 estudios para probar la capacidad del BCB en seleccionar los oocitos más competentes para la producción in vitro de embriones utilizando corderas como donantes de oocitos. Además se estudiará la competencia oocitaria en relación con el citoplasma y factores moleculares, sus respuestas a las diferentes técnicas de fecundación y nuevos medios de maduración para mejorar la PIVE. En el Experimento 1, se analizaron diferentes concentraciones del BCB (13, 26, 39 y 52 µM) y los oocitos fueron MIV-FIV y CIV. Se evaluó: diámetro de los oocitos, actividad mitocondrial, actividad del factor promotor de la maduración (MPF), la expresión de genes relacionados con el metabolismo (ATP1A1 y COX1) y la función constitutiva de la célula (CPEB y S100A10) y el desarrollo hasta el estadio de blastocisto. Los resultados mostraron que 13 µM de BCB es una concentración adecuada que diferencia los oocitos más grandes (BCB+: 123,66 µm), competentes para desarrollar hasta blastocisto (21%) y con más células (69,71±6,19) que los BCB- (106,82 µm, 9% y 45,91±3,35, respectivamente). La actividad mitocondrial fue mayor en los BCB+ que en los BCB- (3369 y 1565 unidades arbitrarias, respectivamente) y también la actividad de MPF (1,479±0,09 y 1,184±0,05 densidad óptica, respectivamente). La expresión de genes no mostró correlación con la calidad de los oocitos. El objetivo del experimento 2 fue mejorar la producción in vitro de los oocitos BCB- mediante la adición de insulina transferrina selenio y ácido ascórbico durante 12 y 24 h de la MIV. El MPF y el contenido de ATP se midieron antes y después de la MIV. Los resultados no mostraron diferencias en la producción de blastocistos entre los oocitos BCB-. El MPF y el ATP aumentaron en todos los grupos (P <0,001) después de la MIV. En el Experimento 3 se estudiaron técnicas de fecundación como la FIV, ICSI y la activación partenogénetica (AP) en los oocitos BCB+ y BCB-. Se analizó el contenido del ATP intracelular. Los oocitos BCB+ tuvieron un desarrollo hasta blastocito significativamente mayor tras la FIV y la AP (31,7% y 20,5%, respectivamente) que los BCB- (6,7% y 8,8%, respectivamente). Sin embargo los oocitos inyectados (ICSI) no mostraron diferencias en los blastocistos producidos entre los BCB+ y BCB- (14,3% vs 11,8%, respectivamente). Los oocitos BCB+ mostraron tener mayor concentración de ATP que los BCB-. Finalmente podemos concluir que el test del BCB es una metodología fácil, rápida y adecuada para seleccionar los oocitos de corderas más competentes. Los oocitos BCB+ mostraron una mayor producción de blastocistos, actividad mitocondrial y del MPF y mayor contenido de ATP que los BCB-. El test de BCB es un método útil para ser incorporado en los protocolos de PIVE cuando se trabaja con un gran y heterogéneo número de oocitos. Sin embargo este test es menos interesante cuando se trabaja con un pequeño número de oocitos, como en laparoscópica o ICSI. / The oocyte quality is used as synonymous of oocyte competence defined as the ability of an oocyte to resume meiosis, cleave following fertilization, develop to the blastocyst stage, induce a pregnancy and bring the offspring to term in good health. The selection of more competent oocytes is an important point in in vitro embryo production programs. The Brilliant Cresyl Blue (BCB) test has been successfully used as a non invasive methodology to classify oocytes according to their cytoplasm coloration as grown (BCB+: blue cytoplasm) or growing (BCB-: colorless cytoplasm) oocytes. To our knowledge there are no previous reports using this test to select competent oocytes in sheep. We have carried out 3 studies to test the ability of the BCB to select the most competent prepubertal sheep oocytes for in vitro blastocyst embryo production. Furthermore, we pretend to improve the knowledge about oocyte competence related to their cytoplasmic and molecular performances, their responses to different techniques of fertilization and to test new maturation media to improve the blastocyst production. In Experiment 1, different concentrations of the BCB test (13, 26, 39 and 52 µM) were analyzed. After BCB culture all of oocytes were IVM-IVF and IVC. The parameters assessed in this study were: oocyte diameter, mitochondrial activity, maturation-promoting factor (MPF) activity, mRNA relative expression (RE) of genes related to metabolism (ATP1A1 and COX1) and constitutive function of the cell (CPEB and S100A10) and the blastocyst development. The results showed that 13 µM BCB during 60 min could be a suitable concentration to differentiate largest (BCB+, 123.66 µm) and most competent oocytes to develop to the blastocyst stage (21%) and with a higher number of cells (69.71±6.19 S.E.M.) compared with non-stained BCB- oocytes (106.82 µm, 9% and 45.91±3.35 S.E.M., respectively). Mitochondrial activity, assessed by MitoTracker Orange CMTMRos probe, was significantly higher in BCB+ than in BCB- oocytes after in vitro maturation (3369 and 1565 Arbitrary Units, respectively) and the MPF activity, assessed by CDC2 kinase activity assay, showed significantly higher activity at metaphase II stage in BCB+ than in BCB- oocytes (1.479±0.09 and 1.184±0.05 optical density, respectively). The mRNA RE of the different genes analyzed in this work did not show a correlation with the oocyte quality. In Experiment 2, the objective was to improve blastocyst production of small (BCB-) oocytes by addition of Insulin Transferrin Selenium and Ascorbic Acid during 12 and 24 h of IVM. MPF and ATP content were measure before and after IVM. Results showed no differences in blastocyst production between BCB oocytes. The MPF and ATP increased in all groups (P<0.001) after the IVM. In Experiment 3, the aim was to test the blastocyst production after IVF, ICSI and PA of BCB+ and BCB- oocytes. ATP content was analyzed. BCB+ oocytes developed significantly higher up to the blastocyst stage after IVF and PA (31.7% and 20.5%, respectively) than BCB- (6.7% and 8.8%, respectively) oocytes. However, ICSI treated oocytes did not show these differences between BCB+ and BCB- oocytes (14.3% vs 11.8%, respectively). BCB+ oocytes had significantly more ATP content than BCB- oocytes. Finally we can conclude that the BCB test is an easy, fast and suitable methodology to select the more competent sheep oocytes. Good quality oocytes (BCB+) showed higher blastocyst production, mitochondrial and MPF activity and ATP content than BCB- oocytes. The BCB test is a useful method to be incorporated in the embryo production protocol where a large and heterogeneous number of oocytes are use to be in vitro fertilized. However, BCB test is less interesting when working with a small number of oocytes such as in Laparoscopic Ovum Pick Up (LOPU) or ICSI.

Group B Streptococcus (GBS), in its transition from commensal to pathogen, will encounter diverse host environments and thus require coordinately controlling its transcriptional responses to these changes. This work was aimed at better understanding the role of two component signal transduction systems (TCS) in GBS pathophysiology through a systematic screening procedure. We first performed a complete inventory and sensory mechanism classification of all putative GBS TCS by genomic analysis. Five TCS were further investigated by the generation of knock-out strains, and in vitro transcriptome analysis identified genes regulated by these systems, ranging from 0.1-3% of the genome. Interestingly, two sugar phosphotransferase systems appeared differently regulated in the knock-out mutant of TCS-16, suggesting an involvement in monitoring carbon source availability. High throughput analysis of bacterial growth on different carbon sources showed that TCS-16 was necessary for growth of GBS on fructose-6-phosphate. Additional transcriptional analysis provided further evidence for a stimulus-response circuit where extracellular fructose-6-phosphate leads to autoinduction of TCS-16 with concomitant dramatic up-regulation of the adjacent operon encoding a phosphotransferase system. The TCS-16-deficient strain exhibited decreased persistence in a model of vaginal colonization and impaired growth/survival in the presence of vaginal mucoid components. All mutant strains were also characterized in a murine model of systemic infection, and inactivation of TCS-17 (also known as RgfAC) resulted in hypervirulence. Our data suggest a role for the previously unknown TCS-16, here named FspSR, in bacterial fitness and carbon metabolism during host colonization, and also provide experimental evidence for TCS-17/RgfAC involvement in virulence.

We performed an innovative systematic meta-analysis of gene expression profiles of whole normal human brain and heart to provide a quantitative transcriptome reference map of it, i.e. a typical reference value of expression for all the known, mapped and uncharacterized (unmapped) transcripts. For this reason, we used the software named TRAM (Transcriptome Mapper), which is able to generate transcriptome maps based on gene expression data from multiple sources. We also analyzed differential gene expression by comparing brain with human foetal brain, with a pool of non-brain tissues and with the three brain sub-region: cerebellum, cerebral cortex and hippocampus, the main regions severely affected with cognitive impairment, as seen in the case of trisomy 21. Data were downloaded from microarray databases, processed and analyzed using TRAM software and validated in vitro by assaying gene expression through several magnitude orders by "Real Time" reverse transcription polymerase chain reaction (RT-PCR). The excellent agreement between in silico and experimental data suggested that our transcriptome maps may be a useful quantitative reference benchmark for gene expression studies related to the human brain and heart. We also generated an integrated quantitative transcriptome map by systematic meta-analysis from all available gene expression profile datasets related to AMKL in paediatric age. The incidence of Acute Megakaryoblastic Leukemia (AMKL) is 500-fold higher in children with Down Syndrome (DS) compared with non-DS children. We present an integrated original model of the DS AMLK transcriptome, providing the identification of genes relevant for its pathophysiology which can potentially be new clinical markers. Finally, computational and molecular analysis of a highly restricted region of chromosome 21, which represents a strong candidate for typical DS features and is considered as intergenic, was performed. Northern Blot analysis and computational biology results show that HR-DSCR contain active loci bidirectionally transcribed.

A seqüência nxh1 foi isolada a partir de uma biblioteca de cDNA da glândula de veneno da cobra coral Micrurus corallinus. Essa sequência apresenta similaridade estrutural com as toxinas de \"três dígitos\", e principalmente com as &#945;-neurotoxinas, devendo apresentar 4 pontes dissulfeto deduzidas por comparação com outras proteínas desta família. Porém, essa potencial toxina não possui alguns aminoácidos importantes para a interação com o receptor nicotínico de acetilcolina na junção neuromuscular. A potencial toxina NXH1 foi expressa em E. coli de 3 maneiras distintas. Os melhores resultados foram obtidos quando a NXH1 foi expressa em fusão com uma cauda de histidina. Essa construção permitiu uma rápida e eficiente purificação da proteína recombinante. A proteína de fusão foi usada para a produção de um soro específico anti-NXH1. O soro anti-proteína recombinante, assim como o soro do Instituto Butantan, reconheceu a toxina recombinante em Western blot e ELISA. Além disso, o anti-NXH1 reconheceu em Western blot apenas uma banda presente no veneno de M. corallinus, mas não reconheceu os venenos de outras 10 espécies de Micrurus. Experimentos de ligação mostraram que componentes do veneno de M. corallinus ligam-se aos receptores nicotínicos das membranas de músculo de rato. O soro anti-NXH1 não inibiu a ligação do veneno aos receptores, indicando que, ou a NXH1 não é uma &#945;-neurotoxina, ou o antisoro não consegue impedir a ligação da toxina nativa ao receptor, ou que existem outras a-neurotoxinas do veneno que não são bloqueadas pelo soro anti-NXH1. / The nxh1 sequence has been isolated from a cDNA library from the coral snake Micrurus corallinus\' venom gland. The deduced protein is highly similar to known \"three finger\" &#945;-neurotoxins, with four deduced disulfide bridges. However, the predicted protein lacks some important amino acids for the nicotinic acetylcholine receptor interaction. The potencial toxin NXH1 was expressed in E. coli in three different ways. The best results were obtained when the protein was expressed as a His-tagged fusion protein, which allowed rapid and efficient purification of the recombinant toxin. The fusion protein was used to generate a specific antiserum against NXH1. The produced antiserum, as well as the serum from Instituto Butantan, recognized in ELISA and Western blot the recombinant toxin. Besides, the anti-NXH1 serum recognized in Western Blot a single band from the venom of M. corallines but not the venom from other 10 Micrurus species. Binding experiments showed that the components from M. corallines venom bind to nicotinic acetylcholine receptor in rat muscle membranes. The anti-NXH1 serum did not inhibited the binding of the venom to the receptors. It seems that NXH1 is not an &#945;-neurotoxin, or that the antiserum does not inhibit the binding of the native toxin to the receptor, or that the antiserum is not capable to inhibit the action of other &#945;-neurotoxin from the venom.

Barreiras reativas permeáveis orgâncias (BRPOs) contendo substratos orgânicos, tais como resíduos agrícolas, podem ser empregadas na remediação in-situ de áreas contaminadas por drenagem ácida de minas, ou de áreas industriais contaminadas por solventes clorados, entre outros. O desempenho da remediação nestas BRPO depende da atividade de bactérias redutoras de sulfato, por sua vez condicionada pela composição e degradabilidade dos materiais orgânicos/celulósicos providos na BRPO. Com base em dados da literatura, são apresentados valores COT, COD, N, ligninha/N, C/N para diferentes resíduos já utilizados como fontes de carbono ou de nutrientes em ensaios em batelada de redução bacteriana de sulfato. A disponibilidade de nutrientes ou de carbono orgânico durável, verificada nos diferentes resíduos analisados, evidencia aspecto de complementariedade e demonstra a importância de trabalhar com misturas em BRPOs. Neste trabalho, foram escolhidos para estudo resíduos agrícolas de grande disponibilidade no Brasil como o bagaço de cana-de-açúcar (o qual foi caracterizado quimicamente) e a cama de aviário. Os ensaios de batelada foram realizados em reatores anaeróbios desenvolvidos a partir de garrafas PET adaptadas com lacre anaeróbio. Este trabalho também apresenta uma metodologia desenvolvida para remoção de compostos orgânicos das amostras coletadas nos reatores e destinadas à análise em cromatógrafo de íons. Os resultados de potencial de oxidação-redução menor que -200 mV, pH (5-7) e variação da concentração de sulfato, indicaram a eficiência do procedimento experimental e da mistura dos substratos bagaço de cana, testado como fonte de carbono durável, e cama de aviário, testada como fonte de nutrientes.

Orientadores: João Lucio de Azevedo , F. G. Brieger / Tese (doutorado) - Universidade Estadual de Campinas. Instituto de Biologia / Made available in DSpace on 2018-09-11T20:49:47Z (GMT). No. of bitstreams: 1 Baracho_IvanhoeRodrigues_D.pdf: 3936659 bytes, checksum: 9e9b9a9c3458de50646c6ba1f7a178fe (MD5) Previous issue date: 1969 / Resumo: O presente trabalho teve por finalidade a realização de estudos correlacionados com a formação de cleistotécios em Aspergillus nidulans (Eidam) Winter. 1. Seis linhagens mutantes de A. nidulans, procedentes do estoque de Glasgov, Grã-Bretanha, foram analisadas com o fim de verificar a variação que apresentavam na produção de cleistotécios, e também para que se pudesse escolher linhagens cleistoteciais e acleistoteciais. As linhagens analisadas foram: (1) y; nic 2; ribo 5 (2) y; w 2; s 12; pyro 4 (3) na 1, bi 1 (4) y, ad 14; co (5) su 1 ad 20, y, ad 20, paba 1; Acr 1; lys 5; cha (6) pro 1, paba 6, y; w 3. Pelos resultados obtidos, apenas a linhagem pro 1, paba 6, y; w 3 pôde ser considerado acleistotecial, com relativa segurança, pois não apresentou produção de cleistotécios em 684 colônias examinadas. As demais linhagens apresentaram produção de cleistotécios, embora essa produção tenha sido irregular e incera, isto é, se bem que para a mesma linhagem sempre se utilizasse uma única colônia, para dessa, se obterem as demais, essas linhagens sempre deram origem a colônias produtoras e colônias não produtoras de cleistotécios. O número de colônias produtoras e não-produtoras de cleistotécios variou em relação à vedação ao não vedação, da placas de petri, com fita celulósica... Observação: O resumo, na íntegra, poderá ser visualizado no texto completo da tese digital / Abstract: Haploid strains of Aspergillus nidulans were used in na attempt to study the variation in the production of cleistothecium. Six strains were analysed. From these strains 5 produced cleistothecia. Erratic variation in the production of cleistothecium was observed. Colonies originated from strains which produce cleistothecium segregate into cleistothecial and acleistothecial forms. The number of cleistothecieal and acleistothecial colonies showed variation under several conditions. A heterokaryon transfer teste indicated the participation of an extrachromosomal factor in the control of the difference between the cleistothecial and acleistothecial states. The genetic control of the formation of cleistothecium is discussed. / Doutorado / Doutor em Ciências Biológicas

A pesquisa apresentada é caracterizada como um estudo de caso, baseada numa investigação empírica que estuda um fenômeno contemporâneo, da sociedade e em especial no ambiente escolar. Foi elaborada uma disciplina para alunos do Ensino Médio de uma escola pública federal, com formato alternativo ao modelo baseado apenas na sala de aula, de caráter eletivo mas como uma disciplina regular. Os dados foram coletados ao longo de 4 semestre nos anos de 2012 e 2013 As atividades foram propostas e aplicadas buscando-se aprimorar o ensino e a aprendizagem de Ciências. A Análise Textual foi a metodologia empregada para interpretação de parte dos dados coletados nesse trabalho. O referencial teórico utilizado pelos pesquisadores para analisar o discurso presente nas informações coletadas é a Educação pela Pesquisa (Demo, Galiazzi, Ramos, Moraes). Essa é a “teoria a priori” que fundamenta a análise na tese, constituindo assim uma Análise Textual Discursiva que assume pressupostos da Análise do Discurso. Assim, a pesquisa objetivou-se na proposição de um método de trabalho onde a observação de aves seja utilizada como instrumento didático para proporcionar o aprendizado dos conceitos e conteúdos de Biologia preconizados nos Parâmetros Curriculares Nacionais (PCNs), suscitando e valorizando a argumentação, a pesquisa e a interdisciplinaridade. A partir da aplicação dessa proposta de trabalho foi feita a análise e avaliação da atividade desenvolvida questionando a sua validade para o processo de ensino e aprendizagem em Biologia. Os resultados são apresentados na forma de artigos e análise qualitativas. Os instrumentos de coleta de informações foram questionários, atividades descritivas e mapas conceituais. Dessa forma, a pesquisa buscou promover reflexões e discussões acerca da elaboração e aplicação de uma metodologia de trabalho para contribuir para a melhoria dos processos de ensino e aprendizagem de Biologia na educação básica. Em função dos resultados e análises desenvolvidos é possível afirmar que houve melhora nos processos de ensino e aprendizagem. / The present research is characterized as a case study, based on an empirical investigation that studies a contemporary phenomenon of society and especially in the school environment. A discipline was developed for high school students of a federal public school, with an alternative format to the classroom, with elective choice but as a regular discipline. The data were collected during 4 semester in the years of 2012 and 2013. The activities were proposed and applied in order to improve the teaching and learning of Sciences. The Textual Analysis was the methodology used to interpret part of the data collected in this work. The theoretical framework used by the researchers to analyze the discourse present in the information collected is the Education through Research (Demo, Galiazzi, Ramos, Moraes). This is the "priori theory" that underlies the analysis in the thesis, thus constituting a Discursive Textual Analysis that assumes the presuppositions of Discourse´s Analysis . Thus, the research was aimed at proposing a work method in which bird observation is used as a teaching tool to provide the learning of the concepts and contents of Biology recommended by National Curricular Parameters (NCPs), raising and valuing the argumentation, the research and interdisciplinarity. From the application of this work proposal was made the analysis and evaluation of the activity developed questioning its validity for the teaching and learning process in Biology. The results are presented in the form of articles and qualitative analysis. The instruments of information collection were questionnaires, descriptive activities and conceptual maps. Thus, the research sought to promote reflections and discussions about the elaboration and application of a work methodology to contribute to the improvement of the teaching and learning processes of Biology in basic education. Due to the results and analyzes developed it is possible to affirm that there was an improvement in the teaching and learning processes.