MICROBIOLOGY (2)

Author: CUEVAS LOBATO OSCAR.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD CIENCIAS QUIMICAS.
Place of preparation: FACULTAD DE FARAMACIA.

Summary: Introduction: The staphylococcal infection is a major problem both in health and in the middle estrahospitalario.La emergence of microorganism increasingly resistant to antimicrobianes commonly used in clinical practice requires action to control the spread of these cepas.El present study makes a description of the current status of Staphylococus aureus in hospitals in Spain as well as the evolution of resistance during the past 20 years. Objectives: They focus on three key aspects: 1.Epidemiología. 2. Resistance antimicrobianoes. 3. Genotypic and phenotypic characterization. Materials and Methods: A prevalence study were collected staphylococci isolated one day in 143 hospitals españoles.Se determined the most important epidemiological parameters (distribution oor type of clinical specimen by size of the hospital, hospital service comes.) All isolates them tests for the identification and sendibilidad to 20 antimicrobvianos commonly used in daily clinical practice, using techniques microdilición in caldo.Asimismo were carried out purebas of phenotypic characterization (fagotipificación and detention of protein PBP2a through aglutianción with partícuals latex) and genotypic characterization (arrest of mecA gene by PCR, characterization of Staphylococcus aureus resistant to methicillin through elctroforesis field and pressed detection cassette cromosomal -SCCmec- through PCR). Results and conclusions: There has been an increase muyn acussdo on the strength of S.aureus to methicillin (1.5% in 1986 vs. 30.5% in 2002). All isolates were uniformly sensitive glycopeptide well com to linezolid, quinupristian - dalfopristin and tigeciclina and low levels of rifampicin resistance and cotrimoxazol.Las techniques tripificación molecular electrofresis field pressed (PFGE) identified 10 clones majority and 22 clones sporadic distributed by todaa national geography, with a current presence of very small "clone Iberian. "criminalization through SCCmec found that 70% of isolates belong to type IV.

Author: CASTILLO LLUVA SONIA.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: CNB.

Summary: The fungus Ustilago maydis, sexual development and pathogenicity are intimately relacionados.Las cells grow as a haploid yeast, divided by gemación, and the induction program requires pathogenicity of mating between two sexually compatible cells that give rise to the formation of a filament dicarionte that invades the planta.En this paper we describe a member belonging to the family Fizzy-related adapter complex APC called Cru1.Este regulator is required to adjust the length of the phase G1 depending on the conditions ambientales.También describe a ciclina of g1, Cln1, has been implicated in controlling cycle regulators involved in the entry into mitosis during G1 / S by activation of inhibitor of the kinase Cdk1, Wee1.Las strains defectivas in cln1 are affected in the ability to infect corn. Finally, we describe the existence of a kinase-related Pho85 S. Cerevisiae submitting a role in the polarized growth of the gem.

Author: GUINEA ORTEGA JESÚS VICENTE.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA . UCM.

Summary: This draft doctoral thesis is divided into three distinct parts that try to answer three questions: 1. On the one hand, we studied the population Gregorio Maranon Hospital, which had a positive culture for A. Fumigatus during a three-year period to assess the likelihood that these patients were not infected. This reflected both microbiological and clinical variables that might predict the presence of infeción if you have a crop pósitivo. We calculated the burden of trabaJo which involves the cultivation of A. Fumigatus in the microbiology laboratory. To get the predictive variables of infection was applied multivariate logistic regression and a ROC curve. Only 22% of the strains recovered in the laboratory micologíarepresentaron enfermePad invasive. Therefore, the overall probability of suilir invasive aspergillosis when a crop is positive for A. Fumigatus, and the predictive value of the crop without any other consideration is low (22.3%). It seteccionaron five variables. Statistical significance (p mayor0, 05) which predicted infection sample obtained by invasive procedures (p = O, 09), "the presence of two or more samples positive correlative" (p = 0.03), leukemia ( p = O, 033), neutropenia '(P = 0.03) and tratamie Steroid (p greater O OOl); these variables are punctuation and assign a final score for each patient was grouped in, 4 categories 0 points, between 1-2 between 3-4 or +-5.Este model gave us the chance to have invasive disease in patients with one or more crop positive depending on the presence or absence of the variables described with a score or they only had a probability of 2.5% of invasive aspergillosis have a rating between 1 and 2 elevation likely to 10.3%, and likely reached 40.% and on the 7th% respectively in those patients with a score of 3-4 - And less 5. 2. A study environmental levels of spores of Aspergillus with objéto will see them levels of spores to be exposed population in the province of Madrid. autumn veriftcaron highest in the other three seasons of año.Los levels of spores of Aspergillus totals were always less than 85 ufc/m3 while for Aspergillus fumigátus these were lower. to 70 ufc/m3. levels spores were influenciarón by meteorological parameters 3. . were evaluated 596 isolated Aspergillus fumtgatus collected from the air. province of Madrid, air Gregorio Maranon hospital and clinical samples of patients admitted to the hospital (in order to study their sensitivity to antifungal and compare the origin the isolated Amphotericin B had the highest WCC half (2.0177 ug / ml), followed by itraconazole (0.8102 ug / rnl), pOsaconazol (0.3853 ug / ml) and voriconazole was the most active (0.3035 ug / ml). equinocandinas (Caspafungina and micafungina) presented some CMEs very low, always mayor0, 006 ug / ml.No significant differences in sensitivity to these antifungals when compared to places of isolation: The incidence of resistance to azole Aspergillus and equinocandinas is very low, Caspofungin and micafungina showed actiVidades antifungal very good.

Author: MARTÍNEZ LÓPEZ RAQUEL.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA. UNIVERSIDAD COMPLUTENSE.

Summary: The protein ECm33p was initially described in S.cerevisiae as a protein GPI included in the family SPS2.Los serious defects present in the cell wall of mutant ecm33 of S.cerevisiae revealed the clear involvement of Ecm33p in the formation of this structure is essential for the viability of the levadura.La identification and characterization of gene homologous to ECM33 in the opportunistic fungal pathogen C. Albicans, has revealed that the protein Ecm33p is involved in addition to the formation of the cell wall, in other important biological processes such as morphogenesis and fungus transition dimórfica.Así, the mutant ecm33 of C.albicans was unable to filamentar media solid as Spider and SLADH generally induce filamentation and invasón of agar.Estos mutants were also totally avirulentos in various in vivo murine model of candidiasis (systemic and mucocutaneous) conducted in this estudio.Mediante in vitro studies showed lower adhesion of mutants ecm33 of C.albicans to both human endothelial cell lines as epiteliales.Así, inoculation of mutant homozygous ecm33/ecm33 (RML2U) C.albicans mice BALB / c was able to induce in them a response immune protective face systemic candidiasis produced by the parent virulent strain of C.albicans SC5314, endowing the mutant RML2U capacity vaccine.

Author: BASSANI CASTELLANOS SYLVINA.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: The HTLV-2 is a retrovirus that infects CD8 + T cells with pathogenic role is discutido.El virus is distributed in the form enfémica in certain regions of America and Africa.En Spain and the rest of Europe, USA The virus has spread among drug users injecting (UDVP), many of whom are co-infected by HIV-1 to be the main reservoir of infección.La influence they can have infection HTLV-2 in subjects HIV-1 is little estudiada.Este paper attempts to analyze the infeccióon by HTLV-2 in various collective risk of Spain, explore the role of inmusupresión produced by HIV-1 in the serologic diagnosis of HTLV-2 and analyze the influence of coinfection HTLV-2 on immunological parameters of infection by HIV-1 and on the natural course of infection with HCV. In conclusion we can say that: Spain, but with regional differences, the prevalence of infection with HTLV-2 is baja.En the 1079 immigrants surveyed, was not detected any cases of HTLV-2.Sólo has recognized a single subtype of HTLV- 2 (b04), in Spain, suggesting that the epidemiological pattern of this infection due to a serological effect fundador.El diagnosis of infection HTLV-2 in patients co-infected with HIV-1 did not seem to engage in subjects that have a deterioration the response inmune.Los patients co-infected with HTLV-2 and HIV-1 filed juvenile HIV viral loads and lower immune activation that patients only infected VIH-1.Esto could mean a lower progression to AIDS in patients co-infected with HIV and HTLV-2.El profile of the immune response is different in both groups pacientes.La infection of HTLV-2 in subjects with HIV, does not seem to alter the natural course of infection with HCV or plasma viremia in chronic phase disease. KEYWORDS: HIV-1, UDI, HCV.

Author: ALVARADO OROSCO NEISA.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA. UCM..

Summary: This draft doctoral thesis is divided into three distinct parts that try to answer three questions: 1. On the one hand, we studied the workload catheters central reperesentan for a general hospital and found that they accounted for 16.54 CVCs for every 1,000 samples total referred to Microbiología.En regard to the CVCs colonized were 13.33 CVCs colonized for every 1.00 incomes and were colonized 4.50 CVCs for every 1,000 samples processed in Microbiology. 2. These were compared prospectively and randomly 3 techniques (Maki, Brun-Buisson and sonication) for the diagnosis of colonization of central venous catheter in 1,000 tips catheter outcome of the overall probability of detection of catheter colonization for each of the three techniques, without taking into account the order in which they were carried out showed that the technique of Maki (88.5%) was higher than Brun-Buisson (76.0% w greater 0.0001), but were not estdísticamente meaningful to compare with sonication (85.4% w = 0314). Moreover, the sonication was superior to Brun-Buisson (p = 0.0001). overall each technique (indistinctly the order in which they were made) techniques Maki and sonication not alcazaron figures statistically significant (p = 1), but both procedures were superior to Brun-Buisson (77.2%) (p = 0012). positive predictive values of the three techniques were (less 97%). 3. We evaluated 2 diagnostic methods for predicting catheter colonization using acridine orange staining of Gram and smear the surface of the catheter with the cultivation in 425 tips catéter.Se found that the values are sensitivity and negative predictive value were higher in Group tinción-cultivo with a 94.3% vs. 64.3% (p greater 0001) respectively stains from the surface of the catheter predict with a high efficiency of catheter colonization, the information may be available in less than an hour and matched with the results of the crop were excelentes.La observation of micro-organisms in smears catheter with a high value predictifo negative and excludes Bacteremia Linked to the Catheter.

Author: ORTEGA DOMÉNECH MONTSERRAT.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE CIENCIAS BIOLÓGICAS.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: We carried out the identification by molecular typing of clones serotype 6B Streptococcus pneumoniae, which are circulating in Spain in recent years, using, isolates of invasive disease, non-invasive and healthy carriers. We also conducted an analysis of the virulence of all clones described through an experimental animal model of sepsis was made that point in the lab. Finally, it got the differential expression of certain proteins involved in virulence, through the application of proteomics technology. It revealed the involvement of clone Spain6B-2 in the production of disease caused by strains of serotype 6B, finding that the strains of origin clinical serotype 6B (invasive and non-invasive) presented a great homogeneity, as most they fall within the aforementioned clone multirresitente Spain6B-2. Moreover, the strains of carrying serotype 6B, expressed great genotypic diversity, as a service, half of them, clones diverse genetic background. Not all strains circulating produce disease with equal frequency. That is why, regardless of the importance it might have capsular types, the genotype of a strain is a determining factor for a single infection can develop. The serotype 6B provides a structure of the population essentially clonal, agrupándose most d strains in different clonal complexes and also develops mainly by recombination processes. Emphasize that the strains of serotype 6B there is evidence of possible recombination processes at the level of operon coding for the capsular polysaccharide. With the mouse model used was found that the c strains belonging to clone Spain6B-2, regardless of the origin of isolation, which are greater virulence presented, which was most significant in the isolated clíncios. The virulent strains reach levels of 10 8UFC/ml blood. However, most of the strains avirulentas not exceed a growth of 10 6UFC/ml. The decline in the fatality rate is a result of the reduction in bacteremia of animals. Therefore, the model of sepsis allows correlate the level of bacteria in blood with the death or survival of the animals. Lastly, note that the two-dimensional electrophoresis could be a good method to detect differential expression of proteins. In a preliminary study with two strains, virulence and avirulencia in the animal model, it was found to give greater expression virulent strain of proteins PspA, PcpA, ClpX and ClpP, all of them allegedly involved in the virulence of the organism.

Author: GARCIA CAMPOS JOSE ANGEL.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA.

Summary: One of the main questions to be answered on Helicobacter pylori infection is to know why only ellO-20% of people infected with this bacteria develop gastric diseases and the rest only develops gastritis. Hence the importance of characterizing clinical isolates from patients suffering from different diseases, with factors associated with virulence. In this thesis we wanted to characterize clinical isolates of H. Pylori from adults or children with ulcer or gastritis, considering the presence or absence of genes associated with virulence classics (cagA, vacA and hpaA); to develop the correct techniques for the study of new genes that are associated with virulence ( hopQ, babA2) or new virulence factors associated with currently is not known exactly what the gene encoding (antigens Lewis). Tambiénhemos wanted to know the sensitivity antimicrobianade these isolates by phenotypic techniques and test the usefulness of molecular techniques for studying resistance claritromicina.Buscamos the link between all these factors. The strains adults are more often positive for cagA gene, hopQ and hpaA. Just as in patients suffering from ulcer is most commonly found cagA + strains, vacAs], hopQ, hpaA and sensiblesa claritromicina.En children are more ffecuentes strains hopQ, hpaA and vacAs2. When grouped strains, taking into account all the factors studied differences emerged between the patterns arising from the strains of patients with gastritis and ulcers, fundamentalmenteen gene expression babA2 and different sensibilidada claritromicinay metronidazole In this estudiono obtuvimosrelaciónentrela patologíay the expression of the antigen Lewis studied. The making of an ELISA is the most sensiblede the comparative method for the detection of the expression of these antigens.

Author: VIÑAS CANALS MARC.
Year: 2004.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGIA.
Place of preparation: FACULTAD DE BIOLOGIA (UNIVERSITAT DE BARCELONA).

Summary: The successive adoption of regulations regarding the contamination of soils RD 9 / 2005, and changes in land use are increasing demand in the decontamination of soil. However, there are still areas in need of research such as soil microbial populations involved in bioremediation processes; studying the fate of contaminants and evaluate ecoxicidad of bioremediation processes. It has carried out a characterization catabolite three consortia microbial undefined, obtained through enrichment processes with different families of hydrocarbons for use in the biorremdiación of soils contaminated with hydrocarbons. They have incubated three consortia (TD, F1AA and AM) with light crude oil (Casablanca). The results show that the ability of the three consortia catabolite is consistent with the substrate used for obtaining it. In addition, the amplification of consortia amid rich for use with actual experiences of bioaumento not diminish their potential degradador. The addition of ramnolípidos MAT10, produced by Pseudomonas aeruginosa AT10 increases the biodegradation of oil by the consortium AM Casablanca, the rate of biodegradation as the degradation of some components such as isoprenoides the fraction saturated and HAPs rented fraction of the aromatic . There has been a characterization of the diversity of microbial consortium AM, degradador of HAPs, using dependent and independent methods of cultivation. This has been isolated strains heterótrofas and degradadoras of HAPs, have been built libraries of cloned gene 16S and 18S rRNA and the population has been studied gene 16S rRNA by DGGE. We have identified a total of 19 components microbial different phylogenetic belonging to the group of Proteobacteria (16/19), Cytophaga-Flexibacter-Bacterioides (CFB) (2 / 19) and Ascomicota (1 / 19). Our results indicate that studies are required MULTIPHASE to learn more deeply the composition of a microbial consortium. It has designed a protocol test biotratabilidad, laboratory-scale, prior to the bioremediation of soils contaminated with hydrocarbons, which consists of 2 phases of the study. The first phase evaluates the presence of microbial populations, their metabolic activity and the actual and potential biodegradability of pollutants in the soil. The second phase studies the optimization of physicochemical conditions (humidity, ventilation, inorganic nutrients, bodisponibilidad) and biological (possibility of microbial populations inoculate allochthonous) that may influence the process of biodegradation for the bioremediation of soils contaminated with hydrocarbons. We have implemented testing biotratabilidad for the bioremediation of contaminated soil and creosote has deepened in the characterization microbiological, chemical and ecotoxicological biodegradation process. The results obtained in Phase I indicate that the soil is suitable for the implementation of the lbiorremdidación. Aeration and humidity of 40% of field capacity have been key factors in achieving a significant biodegradation of TPH and the HAPs of 3 and 4 rings, by the indigenous population in the soil. The analysis by DGGE combined with principal component analysis shows that the structure and composition d microbial communities change in a very different with the addition of nutrients, as well as throughout the process of biodegradation. The bioremediation process decreases toxicity and teratogenicity of leachate evaluated by testing Microtox ® and FETAX, as well as the potential genotoxicity delos TPH analyzed by atomic force microscopy (AFM), but does not diminish the lethality soil entire front to Eisenia foetida.

Author: SECO MUÑOZ CAROLINA.
Year: 2004.
University: PAÍS VASCO [www.ehu.es].
Place of defense: FACULTAD DE CIENCIA Y TECNOLOGIA.
Place of preparation: FACULTAD DE CIENCIA Y TECNOLOGIA.

Summary: Under adverse conditions gram-negative bacteria such as Escherichia coli, entering state not viable arable (VNC). This state VNC includes cells that, while not grow in culture media conventional signs of activity or specific physiological functions. The biological significance and the implications of this state cell have been a subject of controversy and disagreement among microbiologists. Currently we are two interpretations, the state VNC is a strategy to adapt to adverse environmental conditions that adopt some prokaryotes not subject to processes of cell differentiation, or, this state is the result of a degenerative process that leads to cell death. Studies aimed at settling these two options focus on two key aspects: the search for molecular signals involved in the entry and exit of the state and VNC; analysis of the protein composition of the cells in this state as a prelude to identifying the genes involved in the process and as a global knowledge of the molecular phenotype of the state VNC. In the context of molecular signals, our working hypothesis argues that the state transit VNC takes place with the participation of signals célula-célula. These molecular signals would be present in the supernatants from the experiences of survival. In this paper, we study the interrelationships between E. Coli and the chemical composition of the surrounding environment over a process of survival. It extracts obtained free cells in order to characterize chemically and study their impact on populations of E. Coli under adverse conditions and growth in assets. Our results allow us to conclude that during the transition from state to state arable VNC, E. Coli organic molecules released to the surrounding environment although there was no relationship between the excretion of organic molecules and the loss of cultivabilidad. The chemical composition and dynamics of excretion of organic compounds, during transit to the state VNC, varies depending on the environmental conditions instigators of the same. In addition, organic compounds excreted not act as inducers of transit or in cells under adverse conditions, or in cells growing asset. Finally, we believe that the amino acids and carbohydrates excreted by E. Coli under adverse conditions can act as nutrients delaying the loss of cultivabilidad and therefore entry into the state VNC. Moreover, we have characterized the proteome of cultivable cells and cells of E. VNC Coli. In this context, we analyze the relationship between the protein composition of the cells VNC and some environmental variables instigators of that state. To do so, first and foremost, gels were generated pattern.

Author: VILANOVA SOLÁ XAVIER.
Year: 2004.
University: BARCELONA [www.ub.es].
Place of preparation: FACULTAD DE BIOLOGÍA UNIVERSIDAD DE BARCELONA.

Summary: Changes in the structure and composition of the populations of fecal coliform and enterococci along different purification processes of wastewater into five wastewater treatment plants were analyzed in order to identify possible eliminations selective among these strains groups of bacteria. In this regard, were also studied the sub-poblaciones of enterococci resistant to antibiotics erythromycin and vancomycin (VRE and ERE respectively) as a marker to assess its persistence in different purification processes. The choice of these two sub-poblaciones as markers was because of his medical interest. They also discussed the impact of treated water a delas d five plants on the populations of fecal coliform and enterococci River receiver. It also compared the structure and composition of the populations of these indicators bacterial between wastewater and sludge of another of the 5 stations studied. Finally, we compared the diversity and structure of POBLACIONAL enterococci (VRE and including SRE) in wastewater treatment plants, hospitals and water receiving the discharge of treated waste water r, three located in different geographical areas country (Spain, Sweden and the United Kingdom). The diversity and similarity population of fecal coliform and enterococci were assessed through a fenotipado biochemist of a significant number of strains, and its subsequent analysis using statistical correlation studies and grouping. All samples showed high levels of diversity in the bacterial populations studied the structure and composition of these populations was very similar regardless of origin, treatment system or type of sample, as shown by the high rates of population similarity obtained compare different samples. Both the vancomycin-resistant enterococci as resistant to erythromycin was found in all types of shows, with a higher proportion of the populations of ERE. It was also detected high levels of similarity population for enterococci populations studied and the presence of VRE and ERE in all types of shows, when compared water samples analyzed from different countries. There was a selective removal of clones population in sewage for any of the treatment plants studied. Stocks of VRE and ERE persist after the treatment of water purification, as well as the two types of sludge analyzed. These populations were detected both before and after discharge treated water sampled at all points of the river. The persistence of these populations resistant enterococci antibiotics should be seen in programs for wastewater reclamation and disposal or application of sludge.

Author: POTEL ALVARELLOS CARMEN.
Year: 2004.
University: VIGO [www.uvigo.es].
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: FACULTAD DE CIENCIAS UNIVERSIDAD DE VIGO.

Summary: Staphylococcus aureus resistant to methicillin (MRSA) is a variant widely distributed in hospitals around the world. Its resistance to antibiotics beta lactámicos is mediated by the presence of the mecA gene encodes a protein PBP2a. In this paper deals with the study of a collection of 112 MRSA isolated over the period 1997-2001. One strain was studied by analyzing patient clinical factors associated with the presence of MRSA. For the study of the strains were used: A Methods phenotypic A1-Fenotipos to 16 antimicrobial resistance. A2 - Activity mupirocin. A3-Heterorresistencia to oxacillin. A4-Presence of MRSA sensitive to vancomycin and decreased resistance to the same conglomerate. A5 - activity of the test agutinación fast. B-genetic methods. B1-Amplification of mecA gene. B2-amplifying gene mupA. B3 - Detection of the genes aac6 '-aph2 "and ant4'. B4 - RFLP with Alu I Cfo I gene of coagulase. B5 - polymorphic DNA Amplification random, RAPD. B6-PCR Amplification by sequence repetitive REP-PCR. B7-Amplification region X gene spa A. B8 - Sequencing of the gene spaA. B9, Sequencing-seven loci and definition profile alélico, MLST. our midst health measures to take in controlling MRSA include establishing a monitoring system for readmission in patients with a prior MRSA isolation, strengthening control measures in the medical units and extra asepsis in the management of urinary probes and that the partnership between the same and carry a clone epidemic was demonstrated statistically by univariate and multivariate analysis. methods as type fast antibiogram proved to be useful and between genetic methods based on PCR, REP-PCR and RFLP gene of coagulase were suitable for our study population of MRSA. for the identification of clones as belonging to the international pandemic was necessary I used to the sequencing of the gene spaA and MLST. our midst the prevalence of MRSA was 7,2-43% Throughout this study, having been identified as clones pandemic, the Brazilian clone and clone New-York-Japón, and a third clone named D. 83% of the isolates belonged to one of these three clones and 17% clones sporadic. our environment is not isolated MRSA heterorresistentes to oxacillin and vancomycin. mupirocin is effective because it is not isolated strains with high resistance to it. linezolid The new antimicrobial and quinupristina-delfopristina are very effective as it is not isolated strains resistant or sensitive diminished. The presence of a variant of clone Brazilian with the rapid agglutination test negative in which did not identify the capsular polysaccharides and whether the gene cap8 well as the gene spaA and its protein A, is a worrying development since it may not be correctly identified by the microbiology laboratory to pose this as a risk for release. Developments over the years of study indicates a shift in recent clones resistant to aminoglycosides by new clones more sensitive to them. Presumably the latter are better prepared to persist in hospitals. In this paper have not been identified clones whose origin was communautaire.

Author: LANDETE IRANZO JOSÉ MARÍA.
Year: 2004.
University: VALENCIA [www.uv.es].
Place of defense: BIBLIOTECA DEL CAMPUS DE BURJASSOT.
Place of preparation: FACULTAD DE CIENCIAS BIOLÓGICAS.

Summary: Given the negative influence of the presence of amines biógenas in wine, by the adverse effect on human health and the decline in quality of the wine and the presence of lactic acid bacteria (BL) in the same, in this thesis we ask determine the importance of the BL in the presence of these amines in wine, as well as factors that could influence the concentration of the same or at a greater or lesser production by the BL. First, we developed a method to analyze enzyme concentrations of histamine in the production of wine and the same by BL, then analyze the concentration of histamine, tyramine, feniletilamina, putrescina, cadaverina and tryptamine in wines from different geographical areas and different grape varieties, and the influence of pH, malolactic fermentation, fermentation carried out by BL, and periods of storage, plus the ability aminobiogénica bacteria isolated from some wines was also analyzed. Among other results, we saw that it was during malolactic fermentation when increased levels of histamine, tyramine and feniletilamina, but not the other amines studied, in addition BL are capable of producing histamine, tyramine and feniletilamina but not putrescina, cadaverina or tryptamine . Faced with the need to develop a molecular approach, enabling us to identify the presence of the tyrosine decarboxylase gene (tdc) BL home wine, we have the sequencing and molecular characterization of operon tdc of Lactobacillus brevis 9809, leading to this operon contains 4 Complete gene encoding for a tirosin-RNAt synthetase for tyrosine decarboxylase, for a probable tyrosine permease and for a antiporter Na + / H +. Subsequently, we analyzed using molecular methods and biochemical production capacity of histamine, tyramine and feniletilamina by more than 150 strains of BL, in addition to correlate the presence of these amines in wines with the production capacity of the same by the present BL on the same note that Lactobacillus hilgardii and Pediococcus parvulus were responsible for the high levels of histamine into wine and that most of Oenococcus could produce this amine, but not a danger. As for the tyramine, All Lactobacillus brevis and some Lactobacillus hilgardii produced tyramine and feniletilamina attributed to these species responsible for the levels of tyramine and feniletilamina. Finally, we study the influence of physical and chemical factors of wine and the growth phase on the production of histamine, we saw as glucose and fructose and citric acid and malic and histamine decreased the expression of the gene of histidine decarboxylase and both the production of histamine, on the other hand, the histidine increased the expression, the presence of certain levels of ethanol and pyridoxal 5-fosfato increased production of histamine acting on the enzyme histidine decarboxylase, finally witnessing the growth phase of the BL decisively influences on the expression of the gene of histidine decarboxylase.

Author: BAÑOS MOLINA ROSA CARMEN.
Year: 2004.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: One of the features of the bacterial cell is the ability to adapt to environmental changes in the environment in which it takes place by changing the pattern of gene expression. One of the best examples is characterized the response to changing environmental osmolarity, but most work has focused on the study of genes whose expression is induced to high osmolarity. Through random mutagenesis with transposón MudJ will design a strategy for identifying genes in Salmonella enterica serovar Typhimurium whose term would be reduced to high osmolarity. Following this strategy was identified gene yfeR, which encodes for a protein, YfeR described in databases as hypothetical regulator family LysR regulators transcripionales. Members of this family are generally proteins activadoreas of the transcript and are in many genres prokaryotes, regulating genes or operons encoding functions possibility of belonging to the family LysR, was raised as an objective of this study Doctoral Thesis model regulation YfeR . The analysis of sequence of the protein YfeR revealed distinctive features of regulators transcripcionales LysR: a highly conserved N-terminal end with a cominio "helix-turn-helix" binding to the DNA, a chain amino acid 308 residues (proteins LysR have 300 + /-20 Aas), a relationship Lys / Arg anomalous well as homology with both regulators family described as hypothetical as with regulators LysR. By providing the product of the gene yfeR in trans their self-expression is negatively capacity presenting most regulators LysR. The promoter region of the gene yfeR was located union of a sequence of the porteíneas LysR, T-N11-A with Ty A part of a region inverted. By testing delay gel demonstrated the binding capacity of porteína YfeR this region, and therefore the DNA. These results define YfeR as a member of the family LysR. The results of the regulation of gene expression yfeR show both indirectly, through a fusion gene yfeR: lacZ (MudJ), as directly through the trial of protection from Rnasa-ONE, that the gene yfeR is osmoregulado repression at high osmolarity. A 89 pb the start of translation of the gene yfeR was located a pattern of open reading, called yfeH, which trancribía so divergent, characteristics shared by many regulators family LysR and their genes regulated. Thus arose as a working assumption that this was the gene regulated by YfeR. However, the study of the regulation of gene expression yfeH shows that it is induced in stationary phase, but independently of the presence or absence of possible regulatory YfeR and its regulation by osmolarity. Based on this finding raised the possibility that the protein YfeR regulating the expression of other genes adjacent to the alternative. The analysis of cell extracts of a mutant yfeR respect to the wild strain gels 2D demonstrated by the presence of different proteins with differential expression. Two of them could be identified by MALDI-TOF: IbpA, has been implicated in protection from stress by superóxidos and Lrp, a global regulator involved in the modulation of a variety of metabolic functions, as well as genes that are induced in stationary phase and in response to environmental changes. This result positions YfeR as a protein LysR implicated in a web of regulation compared to overall environmental stimuli.

Author: RODRIGUEZ RODRIGUEZ SONIA.
Year: 2004.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA UNIVERSIDAD DE BARCELONA.
Place of preparation: UNIVERSIDAD DE BARCELONA.

Summary: The protein Hha was identified as a modulator of the expression of the toxin alfa-hemolisina in Escherichia coli and belongs to the family of proteins Hha / YmoA involved in fear and osmoregulación of gene expression in enteric bacteria. In addition, Hha interacts with the protein H-NS and both regulate hemolytic operon in E. (hly) Coli. This paper has studied the functionality of the protein Hha through construction of point mutations in the protein and the construction of truncated proteins of the same. In analyzing the ability of these proteins is involved in union with H-NS and therefore Hha would consist of a single domain functional. The previous observation coupled with the known facts on the regulation of hemolytic operon, which H-NS is protein with the ability to join the DNA indicate that Hha performs its function by binding to H-NS or other proteins and not through a union directly with the DNA. This hypothesis is apoa the similarity between the length of the protein family Hha and domain oligomerización the family H-NS and in the presence of conserved boxes along these sequences (also described in this paper). The data obtained through the construction of a protein hybrid between protein Hha and the DNA binding domain of H-NS suggest that Hha (and his entire family) could be equivalent to the N-terminal domain of H-NS, and such hybrid protein is able to supplement some of the phenotypes characteristic of a mutant hns, for example, hemolytic phenotype, the phenotype beta-glucósido or defiencia a mutant hns to grow in a minimal medium supplemented with serine. This complementarity is important dose of this protein hybrid. Moreover, the protein H-NS E. Coli was described more than 30 years ago as a protein associated with nuleoide and presents an important role in regulating global gene expression. H-NS is widely distributed among bacteria Gram.-negativas and has seen its ability to interact with other proteins and protein Cha. In Yersinia enterocolitica regulation of the expression of virulence has been extensively studied, especially the thermoregulation and participates in this process protein regulatory YmoA (counterpart of Hha), although the involvement of H-NS never been demonstrated. This study has identified the gene hns and its corresponding protein, and has raised its importance to the regulation of the expression of virulence factors in Y. Enterocolitica. Data obtained show the interaction between YmoA and H-NS of Y. Enterocolitica implying that the intercción between proteins Hha and H-NS E. Coli is extensible to other members of the two protein families. Mutations in the gene hns have been obtained on a routine basis in different species of bacteria such as E. Coli, salmonella or shigella but nevertheless in this report shows that the gene hns is essential for Y. Enterocolitica. Soló can obtain a mutant hns supplemented in trans (eg with plasmid wild R27 bearer of a orf gene hns). This allows to study the genes affected by the mutation hns in a manner similar to the mutant hns E. Coli.

Author: GONZÁLEZ-ESCALADA MENA M. DEL ALBA.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: Early diagnosis of the disease spread Mycobacterial mainly Mycobacterium tuberculosis, is complicated due to the slow growth of the bacteria in this genre and inespecificidad the clinical and histological. The objective of this study is to evaluate the usefulness of the PCR technique (reaction polymerase chain) in the rapid diagnosis of the disease spread by M.tuberculosis (MTb) and M. Avium (MA). This amplified sequences specific insertion of MTb IS6110 and MA IS1311. Clinical samples were used as biopsies parafinadas bone marrow, since this is one of the main sources of disseminating these microorganisms. Biopsies were from patients with a positive culture or pair Mtb MA and / or histology and / or clinical symptoms suggestive of disease (Group A) were also included biopsies from adult patients (Group B) and children (Group C) without clinical suspicion neither histological Mycobacterial infection spread. The results obtained in different groups of biopsies of this study leads to the conclusion that CRP is not a suitable technique for the diagnosis of the disease spread by mycobacteria in samples of bone marrow, as it is able to detect DNA of these bacteria in both healthy subjects and in patients. Moreover, the presence of mycobacteria genome in samples of bone marrow without histological pathology evident, suggesting the presence of bacilli in a state of dormancy and opens the door to further research to help understand the pathogenic mechanisms of these diseases.

Author: MARTÍNEZ GARCÍA MANUEL.
Year: 2005.
University: ALICANTE [www.ua.es].
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: FACULTAD DE CIENCIAS.

Author: LLORENS BENLLOCH MARÍA AMPARO.
Year: 2005.
University: VALENCIA [www.uv.es].
Place of defense: BIBLIOTECA CAMPUS DE BURJASSOT.
Place of preparation: UNIVERSITAT DE VALÈNCIA.

Summary: Several species of Fusarium gender are important producing Trichothecenes By zearalenone in plants, the most prominent are: Fusarium graminearum, Fusarium culmorum and Fusarium cerealis. These species are important plant pathogens, especially cereal mind, producing them in a variety of diseases that cause severe economic losses to agriculture and food industry. The Trichothecenes By zearalenone relate With disease, micotoxicosis, affecting both men and farm ani ills. His biosíntesis depends largely on biotic and abiotic factors such as temperature, humidity, isolated fungico and substrate plant. The Trichothecenes B include: nivalenol (NIV), deoxynivalenol (DON) yacetil derivatives 3-acetildesoxinivalenol (3-AcDON) and 15-acetildesoxinivalenol (15-AcDON), resulting from episodes of acute toxicity to damage immunotoxic and citotóxicoS. No clear their activity carcinogenica. Zearalenone is a compound heavily estrogenico causing serious problems in animal reproduction, including abortions and infertility. Because of the serious consequences that these mycotoxins cause, the European Community is working on establishing limits of tolerance around them. So this investigaci6n (using techniques cromatograficas and basad as in the DNA) includes: optimization methods analfticos reliable, fast and reproducible for determining Trichothecenes By zearalenone in vegetables. The study of 105 biotic and abiotic factors that condition the growth fungico, yen consequence production of these mycotoxins. The study fisiol6gico and molecular a large number of isolates of Fusarium spp. Potentially producing Trichothecenes By zearalenone, obtained from various food products origin espariol.

Author: MOLINA ARCAS MIRIAM.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA.

Summary: The nucleosides can be modified estructuramente with the aim of generating active compounds in the treatment of solid tumors and lymphoproliferative diseases. These drugs are intemalizados via nucleoside transporters, and the strength of conveyors tumor cells can determine its action citotoxica. Based on this the objective of this dissertation is to analyze the role of nucleoside transporters in sensitivity to antineoplastic drugs derived nucleoside, especially in the treatment of lymphoproliferative diseases. The first part of the study involved the kinetic characterization of transporters present in the cell line derived from leukemia pro-linfocítica JVM-2, as well as transporters responsible for the recruitment of nucleoside analogue fludarabine. Subsequently, the study was extended to the determination of the levels of mRNA, protein and activity of nucleoside transporters present in cells of patients with chronic lymphocytic leukemia. Similarly, measured their sensitivity to fludarabine with the objective of determining whether there was any correlation between transport and cytotoxicity. The data obtained showed levels of protein transporter hent2 may be predictive of sensitivity fludarabine in chronic lymphatic leukemia. To complement the results, a study was conducted with equivalent lymphoma cell lines derived from mantle, with the aim of checking the differential role of transporters equilibrativos in susceptibility to nucleoside analogues depending on the type of disease and drug administered. In this case, it was the conveyor hENT1 which correlacionava with sensitivity to gemcitabine. Thus, taking into account the role played by transporters equilibrativos in sensitivity to drugs derived nucleoside evaluated the role of conveyor hENT1 in the transcriptional response to therapy associated with nuoropirimidinas using microarrays of ADN.Como model was used the tumor cell line derived from breast MCF7, which was initially carried out a characterization of nucleoside transporters present in the line, as well as isoforms responsible for the recruitment of nucleoside analogue 5'-desoxi-5 fluorouridina. The evaluation of the role of hENT1 was performed using pharmacological inhibition of its activity with NBTI.Los changes in gene expression obtained through experiments microarrays of ADNdemostraron that inhibition of hENT1 blocking all or part of the changes trancripcionales associated with the action 5'-DFUR. Therefore, the results obtained show that transporters nucleoside equilibrativos play an important role in susceptibility to antineoplastic drugs derived from nucleosides.

Author: MARTORELL GUEROLA PATRICIA.
Year: 2005.
University: VALENCIA [www.uv.es].
Place of defense: INSTITUTO DE AGROQUÍMICA Y TECNOLOGÍA DE ALIMENTOS C.S.I.C.
Place of preparation: FACULTAD DE FARMACIA Y FACULTAD DE MEDICINA.

Summary: The importance of yeasts known for a long time and with the development of the industry, are used not only for the development of a large number of fermented products (beer, wine, cheeses and sausages), but also for the production of antibiotics , vitamins, enzymes, etc.. However, the yeast also have a downside, because of its potential as alterantes food, 10 involving major economic losses for the industry. In this paper addresses issues related to the identification and molecular characterization of some of the species tipocamente alterante4s belonging to the genera Debaryomyces, Zygosaccharomyces, Dekkera, Pichia and Saccharomyces. The goal is to provide industrial wings of new rapid identification of yeast as well as show, with specific examples, the usefulness of the application of molecular techniques to resolve industry-wide problems. This has desteminado that ocmparicón sequence of the region ribosomal 5.8 S-ITS and the actin gene is the most optimal method for idenatificar a quick and precise species of the genera Debaryomyces, which are involved in altering processed foods. Furthermore, this paper shows how the application of molecular techniques is very useful for determining the yeast alterantes to 10 throughout the cadana food production. In the present work, are two examples of contamination in the industry, particularly in the production of fruits and nougat in the eleboración wine. It identifíco through the technique of mtDNA RFLPs and RAPD-PCR, the kind Zygosaccharomyces bailii was the yeast responsible for the alteration of those nougat, and syrups used to mecerar fruit source of pollution in the cadan ade production. In addition, the physiological characterization of these strains, allowed evidnciar potential alteramnte this yeast, and who have a high resistance to food preservatives, keeping pace with low pH and low aw. Moreover, the use of selective media, com the DBDM as well as the technical analysis of restriction in the region 5.8 S-ITS allowed dientificar species Dekkera Bruxe / Jesnis and Pichia gui / Jiermindii as yeast alterantes more dangerous in wine, for his great ability to produce high levels of 4-etifenol responsible for unpleasant odors in wine. Through the techniques of mtDNA RFLPs with Hinfli and RAPD-PCR was determined that the source of contamination of D. Bruxellensis occurs before aging in casks and that the yeast species P. Gui / Jiermondii are present in the grapes and raspones, 10 coming from outside the cave. Finally, since the determination of the number of yeast is essential for programass deconservación food, it was deemed of great interest to develop a technique that would enable the identification and simultaneous cunatificación yeast directly into the food. In this paper, has developed a protocol for PCR real especifídco for timely S. Cervcisiae utlizando SYBR-Green agent fluorescent. This technique proved to be very sensitive. By allowing the detection of 3-5 CFU / ml in wine. The system developed real-time PCR is also useful for accurately quantify the number of cells of S. Cerevisiae paresentes in wine, prmitiendo thus estimate the risk of disruption of wine durente storage and distribution. In addition, the system can extend to otrosalimentos and beverages where S. Cervisiae pued 8 and cause 2bd alteration.