MICROBIOLOGY

Author: SOLER FALGÁS LARA.
Year: 2003.
University: ROVIRA I VIRGILI.
Place of defense: MEDICINA.
Place of preparation: FACULTAT DE MEDICINA I CIÈNCIES DE LA SALUT.

Summary: Gender Aeromonas comprises baceterias Gram-negative often isolated in the water, but are important to be present in food, and be able to cause diarrhea and infections extraintestinales tables on the man. In this thesis has been shown to be the cause of up to 2% of traveler's diarrhea. The taxonomy of Aeromonas is complex, the genre consists of 15 species difficult to identify with biochemical methods, which is why in this thesis has developed a protocol based on patterns of gene 16S rRNA restriction to identify all species Accepted the Aeromonas. In addition, most clinical laboratories used for species identification methods miniaturized semi other biochemical tests, as demonstrated in this thesis, lead to incorrect identifications of both species as gender, reaching confuse strains of Aeromonas as part of Vibrio. To avoid this confusion has designed a molecular probe, based on the gene coding for glicerofosfolípido-colesterol aciltransferasa and is capable of identifying colonies in Aeromonas primary means of isolation. The correct definition and identification of species that make up the complex phenotypic "A.hydrophila" (A.hydrohila, A.bestiarum. A.salmonicida and A.popoffii) is one of the main problems in the taxonomy of the genus. That is why we have investigated the genetic and biochemical characteristics of this group of species, and it has been shown that the species A.bestiarum and A.salmonicida can not be differentiated by biochemical or genetic methods, suggesting that it is a single species. Another objective has been studying the resistance patterns of the species d eAeromonas to antimicrobial agents. It has been shown that the strains of clinical origin of the species A.hydrophila, A.caviae and A.veronii are sensitive to cephalosporins, aminoglycosides, fluoroquinolones. In addition, it has been shown that quinolones or third-generation cephalosporins should be considered as the treatment of choice for the treatment of traveler's diarrhea caused by Aeromonas. In order to advance the understanding of the epidemiology of Aeromonas is essential to have reliable methods molecular typing. In eta thesis s ehan assessed three technical (RFLP of spacer intergénico 16S-23S, ERIC-PCR and REP-PCR) for the molecular typing of Aeromonas, having demonstrated that all of them are quick and simple. The technique ERIC-PCR has proved to be the most discriminative, but the combination of two of these techniques improves outcomes. The virulence of Aeromonas is complex and multifactorial, and have been described many different genes encoding enzymes (hemolisina, lipases, Dnasas, proteases ..) that have been identified as virulence factors, however there are no studies that investigated the incidence simultaneously of them all in environmental and clinical isolates of all species of the genus. This thesis has investigated the distribution of genes coding for these virulence factors, as well as activiad enzyme they are associated, and environmental isolates of all species of the genus. It has been shown that they are widely present in all of them. Only econtró a statistically significant association between the presence of the genes encoding virulence factors aerolisin / hemolisina and serine proteases and activity beta-hemolítica, suggesting, as other authors have indicated that they would protease and activity beta-hemolítica, suggesting, as other authors have indicated that the serine protease could act as a trigger for the erolisina / hemolis 8 ina. Po 3e2 r útlimo, has been considered of interest to determine whether the species of Aeromonas most relevant in clinical possess the type III secretion system (TTSS), described in other pathogens such as Escherichia coli, Yersinia spp.y Salonella ssp., and whose main function is to bring toxins directly to the host cell cytoplasm. It has been shown that such a system is present in most isolates of A.veronii and A.hydrophila, which shows that these strains have a potential comparable to virulent pathogens mentioned.

Author: ESTEBAN MORENO JAIME.
Year: 2003.
University: COMPLUTENSE DE MADRID.

Summary: Characterization microbiological: It includes a study on the usefulness of a conventional trading system (API-CORYNE) to identify these aislamientos.La initial assumption comtempla the possibility that these organisms may be confused with Gram-positive bacilli difteromorfors.De acueredo with the study, the use of the system could lead to confusion, and not useful for carcterizar microbiologically species estudiadas.En this part also includes the assessment of a technique for total protein electrophoresis (SDS-PAGE) to identify aislamientos.Como concludes the study, the technique is useful which contains strains of reference with which to compare the electroforetogramas. Clinical Significance: It assesses the cases of isolation of these species in clinical samples of patients from FJD from 1979.Se concludes that many strains have that meaning (particularly those from skin and soft tissue), and that the withdrawal of foreign bodies and a specific treatment is essential for the proper development of the sick. Study pathogenicity: It evaluates the ability of intracellular penetration of the isolates available (selected according to their clinical significance) by the technique preiamente described Analysis microcolonias in diploid fibroblasts humanos.De According to our results, the ability of penetration is not related to the ability to produce disease in humans, as only two strains (both from blood cultures) possess such capability. Antimicrobial Sensitivity: First, it conducts a study of sensitivity of the strains through technical reference compared to most antibiotics used to trtar infections caused by these organimos.En Second, it evaluates a different technique (disco-placa) compared to the technical reference, concluding that does not apply to antibiotics bacteriostatic (mainly macrolides).

Author: GRANIZO MARTINEZ JUAN JOSE.
Year: 2003.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: Since 1990 is increased resistance to erythromycin (E) on S.pyogenes and S.pneumoniae and resistance to penicillin (P) in the last kind, being invoked as a cause of antibiotic consumption, but antibiotic resistance indicators (PyE) is not reflected in drug resistance of the family, nor the resistance measured as WCC on the cutting point involves clinical resistance. The pharmacodynamic data implicating macrolides (M) as selectors of resistant strains, so this association between consumption and resistance is not as simple as pointing. OBJECTIVES 1-Describe the evolution of resistance to E.en S.pyogenes and S.pneumoniae and P.en S.pneumoniae from 1986 to 1999 in Spain. 2-Describe consumption My beta-lactámicos in the same period. 3-Establish statistical association between consumption and resistance. 4-Establish the geographical correlation prevalence of resistance in the E in both species. MATERIALS AND METHODS The evolution of resistance was calculated using meta-analysis of observational studies. The consumption of antibiotics will be taken from IMS that evaluates sales survey. The association between consumption and resistance will be established by multiple linear regression (explanatory model) and the correlations between prevalences were measured with the Spearman coefficient. RESULTS There is a linear increase in resistance to PyE in S.pneumoniae, in the period 1986/87 with a reduction in 1998/99. The high resistance to P (WCC increased 2 ug / mL) have a growing trend since 1990, reaching 35%, similar to the resistance to the E (36%). Resistance to the E in S.pyogenes grows exponencialemente with reductions in 1998/99: 30% in 1997 and 21% in 1999. Antibiotics are the most consumed beta-lactámicos, dominating the aminopenicillin (especially amoxicillin) followed by cephalosporins, whose sales are significant since 1991. Penicillins specter of reduced sales presents minimal. The M sales have increased since the launch of M long half-life (IDB, as Clarithromycin in 1989 and OD, as Azitromicina, 1993), at the expense of a reduction in traditional (TID as erythromycin). Multivariate analysis indicates that the resistance to the E in pneumococcus are correlated with M BID and OD and S.pyogenes with M OD. The intermediate resistance to the P in pneumococcus has no association with any drug, high resistance to the consumption of cephalosporin sorales and total resistance to the use of oral aminopenicillin. There are correlations goegráficas in resistance to the E in both espjecies (r increased 0.65). DISCUSSION Long periods window selectors resistant strains in M largba half-life and cefalosproinas, coselección resistance aPyE in nuemococo and coresistencias to E in both species suggests mechanisms external to the bacteria as selectors resistant strains. CONCLUSION The arrival in the market for M long half-life (IDB) in 1989/90 coincide with the increase in resitencias to EyP in S.pneumoniae and E in S.pyogenes.

Author: MENDEZ MONTESINO JOSE RAMÓN.
Year: 2003.
University: COMPLUTENSE DE MADRID.

Summary: This thesis is based on a descriptive study of hospital setting, which allows to observe and represent the consumption of antimicrobials in the hospital and to determine whether there are differences in consumption in the various hospital services and time trends from 1993 to 2002 inclusive . This thesis is mainly an attempt to determine the grounds for establishing the indicators that should be used to be able to adequately consider the use of antibiotics, with emphasis on trends in the use of the same over the period studied. Studies pharmacovigilance can serve as a management tool because it allows you to control the political use of antibiotics in hospitals. The discrepancy between the health and activity of antibiotics consumption shown in this thesis should be analyzed in future studies.

Author: PEREIRA ARMADA SUSANA.
Year: 2003.
University: VIGO.
Place of defense: SALON DE CUADROS.
Place of preparation: FACULTAD DE CIENCIAS DE VIGO.

Summary: This report deals with the feasibility study bacterial laboratory under controlled conditions, and in the marine environment. This would have employed various techniques microscopic, citométricas and culture. 1-Determination of the total number of cells: acridine orange staining with, the use of cameras and counting staining Kit Live / Dead BacLight. 2-Determination of biomass viable method Kogure (extent of the incorporation of substrate and cell division), reduction or CTC INT (detection of respiratory activity), staining kit Live / Dead BacLight (existence of selective permeability membrane ) and planting solid growth media. Studies in the laboratory have allowed characterize responses survival Listonella pelagia and Vibrio splendidus biotype 1. Both species: 1, Survival long-term (more than 3 months) in microcosm in conditions closer to those in the seawater (low concentrations of nutrients and salinity of 33 per thousand), in this state, L . pelagia significantly reduces its size and becomes cocoide, raising a band of outer membrane protein of 36 kDa. 2, non-viable state arable (VNC), led salinities minors 16 per thousand and / or a concentration of nutrients of 17 g l-1. The cells VNC of L.pelagia are cocides may occur or not increased biovolume. No changes occurred in the profiles of proteins or membrane in lipolisacáridos of the cell wall. L.pelagia recovered "in vitro" (after incubation with ampicillin concentrations bactericides and penicilinasa) and "in vivo" (after injection into gold) 2 / 19 cases. The absence of a sequence of loss of cellular activities prior to cell death, pointing to the state VNC as a survival strategy for these species. Studies with bacteriplancton of the Ria de Vigo have detected the existence of physiological uan great heterogeneity: 1 - Bacteria total abundances showed typical topsoil marine illuminated (3.7 x 10 5 to 5.0 x 10 6 cls ml - 1), being constituted, on average, for a 32.63% of stocks autotróficas (presumptively cyanobacteria of the genus Prochlorococcus). 2-A 48.9% of the total population presented a membrane intact and a 12.68% respiratory activity, with a low percentage of arable AM (0136%) or TCBS (0013%). 3 - The number of total bacteria, total heterórtrofas, bacteria with intact membranes and respiratory activity was highest in July and September, while counts in marine agar were maximum in November. One of the populations showed a pattern of cyanobacteria temporary opposed to presenting the total bacteria counts. 4 - had no detectable variation in the abundances between different sampling stations and depths studied. The bacteria counts and total bacterial membrane intact and respiratory activity was positively correlated with the dissolved oxygen and pH.

Author: URQUIA GRANDE MARIA LUISA.
Year: 2003.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: POLICLINICA CGEA FACUTAD DE FARMACIA.

Summary: The widespread concern about the harmful effects of overuse of antibiotics, has been based on the relationship between consumption and the emergence of resistant pathogenic bacteria but oral streptococci of flora also have developed forms of resistance and may serve as an indicator of the pressure selective of antimicrobianos.Los young adults constitute segment is smaller. A 7.5% had used any antibiotics in the last mes.Se observed a dencenso in consumption over the years, 1991.1995 and 2002. We ordered CMIs of oral streptococci, three antibiotics: penicillin, erythromycin and ciprofloxacin, representatives of the three groups plus antibiotics consumed by the general population, B-lactámicos, macrolides and quinolonas.El 26% of the streptococcus group viridans of 330 young healthy age 25 + -5años are resistant to penicillin (CMImayor least 0.25 ugr / ml). impact of individual consumption of penicillin is remarkable, CMIs to penicillin were significantly higher in the group of young who had taken a B-lactámico in the last month, than the group not tomaron.No have found correlations between the resistance shown by oral streptococci to antibiotincos unrelated as erythromycin, penilicina and ciprofloxacin. The transfer of resistance genes to other related species, as S.pneumoniae turns S.mitis, the most abundant species in the flora oral (60%) in potential diseminadora resistance.

Author: PÉREZ VÁZQUEZ M. DOLORES.
Year: 2003.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA.

Summary: Background: Haemophilus influenzae is a community pathogen that causes acute and chronic infections, invasive and non-invasive humano.La be in the resistance to various antibiotics families is a common problem in clinical isolates Spanish Haemophilus, and the recent emergence of strains resistant fluorquinolonas , the most potent antibiotics known face Haemophilus and other pathogens, poses new challenges in terms of clinical, epidemiological and scientific. Objective: To determine the patterns of microbial resistance and / or clinical fluorquinolonas and other families of antibiotics, population distribution of resistant strains and macanismos resistance (amendment of the targets and others) of Haemophilus influenzae clinical isolates from different pathological processes with decreased sensitivity to ciprofloxacin. Methodology: Survey laboratory strains actively studied to discard loss of sensitivity to ciprofloxacin and / or acid by the Reference Laboratory Haemophilus LRH Center's National Microbiology in the last 7 años.Las strains of clinical origin, have been well characterized by the LRH and collaborators from different hospitals throughout Spain in general, and in particular U.S., Ramon y Cajal Hospital in Madrid in a line of collaboration and research in pathogen strains Fibrosis Quística.El group study comprises 34 strains resistant to quinolones but an equal number of controls sensitive, selected according to the pathology and origin geográfica.Métodos used: (a) Molecular Epidemiology of all strains using markers populations PFGE, (b) phenotypes and resistance patterns 12 quinolones of different generations through dilutions of antibiotics and obtaining CMIs and statistical analysis  © magnification of the regions associated with resistance at gyrA, parC, gyrB and parE according to the literature and data from the genome sequence Haemophilus (d) transformation genetic sequences modified (e) Automatic sequencing of genes and wild transformants and comparison and alignment informástico of them (f) búsquedca other mechanisms possibly involved (pump flow amended)

Author: RADA MARTÍNEZ RAMÓN.
Year: 2003.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA - UNIVERSIDAD COMPLUTENSE DE MADRID.

Summary: Urine samples were collected and vaginal exudate pregnant and non-pregnant women in the same age range for a period of twenty months in a health area of Albacete. The investigation consists of three stages: 1Â fourth cross-sectional study of samples of urine and vaginal exudate pregnant women. 2 Â º Case-control study (versus non-pregnant pregnant) of urinary tract infection. 3 Â º Case-control study of genital infection. We studied the reliability of the test strip and urinary sediment over the uro-cultivo (gold standard) for the early diagnosis of urinary tract infection in pregnant women. It established two logistic regression models, one based on the variables of the test strip, and another from urinary sediment, escogiéndose the established based on the variables of urine sediment for better predictive power of positive urocultivo considered risk . Likewise, with the data obtained from the above model, created a scale clinical risk and a diagnostic algorithm, with similar predictive power. It also studied the prevalence of bacteria in the urine samples of pregnant women. There were statistically significant differences in prevalence between the two groups of samples (urine of pregnant women versus urine of non-pregnant) of the isolates of E. coli, Klebsiella penumoniae, beta-hemolytic Streptococcus agalactiae, Streptococcus agalactiae not hemolytic, E.faecalis and S.epidermidis. In the patterns of sensitivity to antimicrobial most frequently isolated as etiologic agents of infection in crops urine of women embarazadazas and non-pregnant women, there was a statistically significant difference in the percentages of sensitivity to quinolones and gentamicin of S.epidermis isolated in the urine cultures of the two population groups. There was no statistically significant difference between the percentages of antimicrobial susceptibility testing versus Streptococcus agalctiae beta-hemolytic isolated vaginal exudate in pregnant women (cases) and non-pregnant women (controls).

Author: HERRERA LEON LAURA.
Year: 2003.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE CIENCIAS BIOLÓGICAS.
Place of preparation: FACULTAD DE CIENCIAS BIOLÓGICAS.

Summary: The appearance and control of multiresistant strains of Mycobacterium tuberculosis is a very topical issue and the utmost importance Health Pública.El laboratory National Reference Microbacterias, performs a continuous monitoring of these strains obtaining overall figures of resistance from 7.5 % and 3.5% of multiresistant strains (strains resistant to at least isonizida and rifampicin). Currently there are molecular methods for studying the genetic bases of the resistance, to analyze the variability of genes involved in resistance to rifampin (rpob) and Isoniacida (katg, inha / maba and oxyr-ahpc) with which we can detect the mutations, we can develop rapid methods of detection so that the time required for the detection of resistance to these two drugs are shortening, which may establish an appropriate treatment regimen while avoiding the spread of resistant strains.

Author: HERRERA LEON SILVIA.
Year: 2003.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE CIENCIAS BIOLÓGICAS.
Place of preparation: FACULTAD DE CIENCIAS BIOLÓGICAS.

Summary: The bacteria of the genus Salmonella serotypes are classified in traditionally have been a particular association antíganos somatic "O", encoded by the cluster rfb and flagellar antigen "H", encoded by genes fliC and fljB.Actualmente its determination takes by agglutination witha necessarily extensive collection of sueros.El knowledge of the sequences of these genes can discover the genetic polymorphism of each of the alleles codificann for these antigens and thus enables design methods based on the chain reaction polymerase that allow for serotyping molecular bacteria of this genre. Objectives: To contribute to the development of rapid methods for serotyping of Salmonella. 1. Determine the sequences of allele fliCb, fliCd, fliCe h, fliC g, m, fliC i, v, and fliCz10 gene fliC and the alleles fljB1, 2fijB 1.5, fljB1, 6, fljB1, 7, fljB e, n, x, fljBe, n, z, 15, and fljB i, w gene fljB Salmonella flagellar antigen encoding of 1Â th and 2Â Stage of the most common serotypes. 2. Determine the consensus sequence domain central described these alleles for the definition of a genetic polymorphism among them. 3. Design initiators specific PCR to identify these alleles. 4. Compare the results obtained with the traditional serotyping

Author: ROMÁN ALONSO FEDERICO-EMILIO.
Year: 2003.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE CIENCIAS BIOLÓGICAS.
Place of preparation: FACULTAD DE CIENCIAS BIOLÓGICAS.

Summary: H. influenzae is one of the first pathogens responsible for infectious morbidity of patients of cystic fibrosis (CF). We propose to contribute to the improvement and advancement of knowledge about the significance of H. influenzae in CF. We carried out a prospective study of 30 patients with CF over a long period of time to determine whether these patients maintained the same strains of H. influenzae or were subsecuentamente colonized by various other, to study the profiles of antimicrobial sensitivity compared to patients without CF, in order to discern whether antimicrobial resistance and persistence were associated, to check whether existína high proportions of strains hipermutables Patients in CF and sequence the gene mutS of strains hipermutables. We got 188 isolates belonging to 115 genotypes other than H. influenzae. All strains were not capsuladas except 4 which belonged to serotype F. Note changes of outer membrane proteins in 9 of these genotypes. We demonstrate the persistence of the same clone for years in almost 12% of patients. Colonization chronic maintained for years is associated with an increased resistance to antibiotics. The strains of H. influenzae from CF patients are much more resistant to the antibiotics that you strains of patients without CF. At the beginning of the work we detect and study the first cases of patients with CF strains of H. influenzae resitentes to cirpofloxacina and secuenciamos genes gyrA and parC of these strains. The strains of CF showed a high prevalence of decreased sensitivity to ciprofloxacin, among the highest in the literature (21.3%). We describe for the first time the existence of a significant proportion (14.5%) strains hipermutables of H. influenzae in patients with CF. The strains hipermutables are more resistant to antibiotics. The analysis d elas ecuencias gene mutS in 17 of the 19 strains hipermutables of H. influenzae showed the presence of mutations and deletions complex.

Author: PELAEZ ROS BEATRIZ.
Year: 2004.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: ASSUMPTIONS New technologies fail to the efficacy of ethylene oxide and increasing its effectiveness is linked to an increase in the cost of sterile product, therefore, do not constitute a global alternative to ethylene oxide sterilization. Objectives To determine the eficaica microbiological, cost-effectiveness and cost-utility of new technologies sterilization respect to ethylene oxide. Design: Experimental in vitro microbiological efficacy. Survey cost and cost. Scope of the study: Clinical Hospital San Carlos. Central Sterilization Laboratory S.de and Preventive Medicine. Teams sterilizers under study: Ethylene oxide (EO), low-temperature vapor and formaldehyde (VBTF) and Sterrad 100S (S100S) MATERIALS AND METHODS microbiological efficacy was determined (logarithms 10 reduced regard to positive controls in different models instrumental) in the presence and absence of substances interfirientes .. We calculated the cost per 100 l for each method. For the study coste-utilidad analyzed the cost-effectiveness, valuing the indirect costs during periods of availability of sterile equipment and the costs of toxicity. RESULTS Both cycles VBTF showed that more effective microbiological OE and S100S. In experimental models without lúmen and fair conditions were not encontaron statistical difference between the methods evaluated. In the presence of blood on S100S proved less effective in models with lumens, evideciándose earmarking systems S100S and OE for the presence of salt. The sterilization system that cost less per 100 l showed was VBTF. The S100S proved to be the most expensive. CONCLUSIONS VBTF has been the most efficient method, and the cycle of 60Â ° C method of sterilization cheaply. Therefore, it is a valid alternative to comprehensive sterilization traditional ethylene oxide.

Author: FERNANDEZ SESTELO ANA BELEN.
Year: 2004.
University: SANTIAGO DE COMPOSTELA.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE BIOLOGIA.

Summary: This research work has been structured into chapters. There is a general introduction on proteases and three chapters in which reference is made to the study of the same in the three separate microorganisms. The proteases are enzymes degradativos catalyzing the breakdown of links peptídicos of proteins. They appear in a wide variety of natural sources such as animals, plants, microorganisms, protozoa and viruses. The study of microbial proteases is increasing due to its importance in industrial application and biotechnology. The overall objective of this work is to búsquedo of microorganisms producing proteases with potential industrial application. Based on the ability of Candida caseinolytica to produce a protease interest biotechnology we ask your purification and characterization, the study of cell growth and optimizing its industrial production in a fermentor and its application in the digestion of fish remains. After conducting experiments designed for such purposes, we reached the following conclusions: Candida caseinolytica produces an enzyme activity caseinolitica of 35 kDa molecular weight with an optimum temperature of 30Â ° C and a wide range of pH optimum. The results of scaling in a fermenter 20 L capacity showed that its industrial application is feasible. By HPLC analysis of the samples obtained after digestion of protein remains of fish with concentrated enzyme concludes that it is a endopeptidasa. Yarrowia lipolytica secret high levels of extracellular enzymes. The protease is the most studied extracellular protease alkaline (AEP), which is encoded by the gene xpr2. The main objective raised in this chapter was subclonación and overexpression of the gene xpr2 in Pichia pastoris. So came to the following conclusions: The gene xpr2 of Yarrowia expressed in the Pichia pastoris strain GS115 of using the vector pHIL-D2 Through Southern is found that the genome of transforming Pichia has inserted a copy of the gene xpr2. By Northern found that the recombinant DNA is correctly transcribed mRNA The recombinant protease differs as to the optimum pH (pH 7.5) of the native enzyme (pH 9). When cultivated strain transformed into a means of milk production takes place in the absence of protease inducer. The USC Bacillus licheniformis strain used in this study has been aisalada and characterized in our laboratory after a search of strains with milk clotting activity. The purpose of this chapter is to obtain masses queseras using the enzyme released by the strain USC. Once developed experiments concluded: Bacillus licheniformis USC showed clotting activity on skimmed and whole milk. The enzyme is inhibited concentrate entirely by PMSF suggesting that its core asset is rich in serinas. By purifying the enzyme concentrate distinguishes two activities proteásicas; one of 62 kDa with a pH optimum of 7.5, optimum temperature of 37Â ° C pl of 9.2, and another 35 kDa pH optimum of 6.0 and temperature optimal 30Â ° C. The USC strain can be used as a potential new source of enzymes coagulants with industrial application in the development of masses queseras. A preliminary study of the masses queseras obtained shows that the properties físico-quimicas improved significantly with respect to the masses queseras developed with cuajos trade

Author: LASALA SÁNCHEZ FATIMA.
Year: 2004.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FAC. MEDICINA U.C. DE MADRID.

Summary: This paper describes a new mechanism in pathogenic infection by the Ebola virus which is the major use of cell receptors, DC and L-SIGN, for the entry of the virus into the cell and its spread throughout the body. DC-SIGN is a molecule membrane, exclusive of dendritic cells that seems to play an important role in the process of activation of T lymphocytes that they have a high affinity for certain pathogens, among which was gracious Human Immunodeficiency Virus. Using different test systems demonstrates the interaction between the lectin DC and L-SIGN and the Ebola virus glycoprotein, which serves as the basis for caracterizaicón phenomenon and evaluation of this mechanism as a therapeutic target.

Author: RODRÍGUEZ CAPRILES CLAUDIA IRENE.
Year: 2004.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA - UCM.

Summary: JUSTIFICATION The relative incidence of fungal infections in transplant patients has increased in comparison to those of bacterial or viral infections, which have been reduced since there are effective preventive measures against them. Among patients with heart transplant, Aspergillus is the opportunistic pathogen with the highest mortality, however, prophylaxis ideal has not been set. OBJECTIVES Identifying the group of patients who would benefit from prophylactic antifungal directed and set the tone for this administration optic prophylaxis. METHODS were analyzed factors reisgo for invasive aspergillosis in patients who received a heart transplant at our center between 1988 and 2002, as well as the role of oral prophylaxis with itraconazole, which was universally administered from 1995. During the study period 307 patients received a heart transplant, of which 278 have survived more than a week and were included in the analysis. There were 24 cases of aspergillosis. RESULTS The analysis univariables, risk factors for invasive aspergillosis were active infection before transplantation (11% vs. 16.7%, p = 0.03), COPD (5.5% vs. 12.5%, p = 0.02) hyperbilirubinemia before transplantation (greater 2 mg / dl) (40.2% vs. 62.5%, p = 0.3), transfusions depsués transplantation (66.5% vs. 87.5%, p = 0.03), mechanical ventilation long (2.9 vs. 5.4 days, p = 0.05), the use of OKT3 for induction (5.5% vs. 16.7%, p = 0.05), hemodialysis (5.1% vs. 20.8%, p = 0.01), diabetes mellitus (25.6% vs. 50%, p = 0.01), higher number of bacterial infections in the first three months (0, 6 vs. 1.1, p = 0.01), viral infections in the first 3 months (46.9% vs. 75%, p = 0.01), CMV disease in the first 3 months (14.6% vs. 45.8%, p = 0001), and the existence of another case of aspergillosis happened in the area of transplantation caradíaco in do smeses before or after the date of the transplant (47.2% vs. 75%, p = 0 , 01). The prophylaxis with itraconazole was a protective factor (35.4% vs. 8.3%. P = 0.01). The multivariate analysis showed that the risk factors were independently produced invasive aspergillosis: re-operación (RR 5.8, 95% CI 1,8-18, p = 0002), and the existence of another episode of invasive aspergillosis 2 months before or after transplantation (RR 4.6, 95% CI 1,5-14,4, p = 0007). Itraconazole prophylaxis had an independent protective value (RR 0.2, 95% CI 0,07-0,9, p = 0.03). The curve analysis Cox showed an increase in survival in patients receiving itraconazole (RR 0.5, 95% CI 0,3-0,8, p = 0.01). Once established risk factors, we see that 41% of our patients did not present any of them, and consequently, they were not needed prophylaxis. Of the 24 patients with invasive aspergillosis, 23 had at least one risk factor. The episodes of invasive aspergillosis occurred in a period of 0 to 44 days after the beginning of any of the risk factors identified in those patients in whom happened after the resolution of the predisposing condition, the diagnosis is in the period of 30 days. CONCLUSIONS itraconazole prophylaxis should be offered to patients with at least one of these risk factors, iniciandola upon detection and maintained while now and at least in the month following its resolution.

Author: VALER LÓPEZ-FANDO M. LUISA.
Year: 2004.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE FARMACIA.
Place of preparation: FACULTAD DE FARMACIA.

Summary: The protease inhibitors (PI) in combination with other antiretroviral drugs have significantly changed the natural history of HIV infection. Due to the poor pharmacokinetic characteristics of IP, they should be administered along with small doses of ritonavir in order to increase the plasma levels of the same, thereby improving its antiviral activity. Rescue treatments that include enhanced IP are a tool of choice after one or more failures. These combinations have been shown to be safe and effective. The determination of parameters such as genotype in protease patient before starting the new treatment and plasma levels at 12 weeks of treatment, had a value predicitivo in response to it.

Author: ANGULO BURGOS M. ALEJANDRA.
Year: 2004.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: F. MEDICINA.

Summary: The portal hypertension (HP) secondary to obstruction of blood flow portal is one of the complications developed in patients with chronic liver diseases, which are a major cause of death among adults worldwide. Moreover, it is the main complication of liver cirrhosis and is responsible for its consequences more serious rupture bleeding varices esofagogástricas, ascites and encephalopathy protosistémica. A better understanding of the pathophysiological mechanisms of the HP, as well as their complicaicones allow the development of effective methods of prevention and treatment to reduce the high morbidity and mortality of this condition. In this paper we studied the impact of the increase in portal venous pressure in the shaft hepato-intestinal both in the short and long term. We have found that the alterations observed from a qualitative point of view, are the same in the short and long term, while quantitatively differ suggesting that this is a progressive process seems to be starting bowel. Changes intestinal behave impact liver targeted for a change in the hepatic metabolism of lipids. In conclusion, portal hypertension induced alteration inflammatory shaft hepato-intestinal, which evolves as a long-term process of remodeling, as in other chronic inflammatory diseases.

Author: MAZÓN BUSQUETS GERARD.
Year: 2004.
University: AUTÓNOMA DE BARCELONA.
Place of defense: FACULTAD DE CIENCIAS.
Place of preparation: ESCUELA DE POSTGRADO.

Summary: The SOS system is a network multigénica that is induced by the presence of lesions in DNA. It consists of genes involved in replication, repair, mutagenesis and control of the cell cycle. This network multigénica has been characterized in different species of bacteria, gram-positive and gram-negative determined for all of them why their union repressor, the protein LexA. The present paper describes the characterization of the reason for joining the protein LexA in the microorganism Xylella fastidiosa, Anabaena sp.y Fibrobacter succionogenes. By searches with the program TBLASTN, genes lexA of these bacterial species have been identified and cloned into the vector to produce and purify through affinity columns protein LexA encoding. The purified protein has been made tests and mobility electroforética "footprinting" in determining their union sequences for protein LexA, which have been defined how: TTAGN6TACTA for X.fastidiosa, RGTACNNDGTWCB for Anabaena and TGCNCN4GTGCA for F.succinogenes. The reasons for union characterized has served to define the composition of regulones LeX in these microorganisms and demonstrate the presence of inducible genes before the damage laADN independently of LexA in X.fastidiosa. The comparison of the reasons for joining LexA characterized in this work with those previously described in the biography, directed mutagenesis study by the motivios and analysis phylogenetic LexA and RecA, have enabled us to reconstruct the evolutionary history of the reason for joining LexA and the presence of a discrepancy in the cause of unity of the Proteobacteria Alpha, demonstrating that its gene LexA has been incorporated by horizontal exchange from a bacterium related cinobacterias.

Author: OCHOA CASTILLO CAROLINA.
Year: 2004.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD CIENCIAS BIOLOGICAS.
Place of preparation: LABORATORIO LETI S.L..

Summary: The allergic sensitization versus domestic mite allergens is a problem whose incidence is increasing in most industrial countries ised. Although the main species at the global level is Dermatophagoides pteronyssinus, recent data suggest the importance alergologica of Blomia tropicalis, especially in tropical countries and subtrópicales where it can be seen from greater importance equaled D. Pteronyssinus. To date, they have been purified and sequenced several allergens B. Tropicalis, usually from extracts of bodies. However, despite all that has recognized the importance of allergenic excreta, has not been studied in detail the composition of allergenic extracts fecal. Our goal were to characterize inmunoquimicamente extracts somatic and fecal B. Tropicalis, studying its antigenic composition, allergenic and enzyme activity; study the cross-reactivity between the extract somatic and faecal B. Tropicalis other species of mites that cause allergy and with a kind of cockroach; study the kinetics of quick-release using different solvent, purify a protein using conventional technology. There have been qualitative and quantitative differences between the two excerpts (somatic and manure) in terms of the pattern and allergenic protein. It detected 32 bands joining IgE in the excerpt somatic and 29 in the extract manure, in a range of 9.2 to 95.5 kDa. There were also differences in enzyme activity was detected between the two excerpts minimum cross-reactivity with D. Pteronyssinus, D. Farinae and Euroglyphus maynei, remain high with Tyrophagus putrescentiae and Lepidoglyphus destroyer especially with the extract of fecal B. Tropicalis. There is cross-reactivity between the extract somatic B. Tropicalis and cockroach Blatella germanica The kinetics of release of allergen showed similar dynamics using different solvents (water, PBS NaCI 0.85%), demonstrating the high solubility of proteins, particularly those of a 14 kDa in volume of rápída appearance. By ammonium sulfate fractionation and hydrophobic interaction chromatography, semipurificamos proteins around 14 kDa, with a frequency of recognition by sera from patients allergic to a 24.3%.

Author: ROMERO FERNÁNDEZ PATRICIA.
Year: 2004.
University: COMPLUTENSE DE MADRID.
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FAC. BIOLOGÍA (UCM).

Summary: SUMMARY: Streptococcus pneumoniae is a major human pathogens. In children is primarily responsible for otitis media and one of the most important cause of meningitis and pneumonia. In adults is the major causative agent of the community-acquired pneumonia, as well as for sinusitis, meningitis and bacteremia. Its ecological niche is the respiratory tract of human beings and thus it is considered that any individual has been colonized at some point in their lives. Streptococcus mitis is a species with which it shares its natural habitat and is very close in terms of taxonomy. Proper identification between them is crucial because, whether an isolated pneumococcus is kind or another group mitis determines the treatment to prescribe. Most of the clinical isolates of S. Pneumoniae are lisogénicos meaning that carry fagos inserted into its genome. The presence of these profagos could be an important contributing to the virulence of these important human pathogens. In the work done in this thesis has been made complete sequencing of profago EJ-1 (belonging to the family Myoviridae) isolated from a clinical strain that initially was identified as an atypical pneumococcus and, finally, has been assigned to the kind S. Mitis. EJ-1 is the first of phage family Myoviridae that infects bacteria with a low content of G + C which has been sequenced in its entirety. The detailed study of its genome revealed a gene organization in the mosaic and that only genes encoding proteins responsible for the tail shows a similar significant homológos in fagos Myoviridae while the rest of orf show similarity to genes fagos Siphoviridae . Besides profago EJ-1, in this thesis has been studied two new strains lisógenas S. Mitis. Strains 86 and HER are two clinical isolates of S. Mitis carrying fagos tempered. The fagos B6 and HER encoded in their genome amidasas type LytA, a virulence factor typical of pneumococcus. After detailed study of these enzymes, it was discovered that at position 317 with a residue Thr Val instead of what presents amidasas LytA both bacterial and fágicas pneumococcal studied so far. The construction and the study of a collection of 21 mutants of the enzyme LytA has revealed that the Vab17 and to a lesser degree, aa Tyr294 and Leu314 play a key role in the process of conversíón / dimerización of amidasa needed to achieve how totally active in the main autolisina of pneumococcus.