SOARES, Fábio Alves. Análise da freqüência de anorretocele em mulheres adultas com evacuação obstruída, comparando com a paridade e idade, utilizando cinedefecografia e eletromanometria anorretal. 2006. 66 f. Dissertação (Mestrado em Cirurgia) - Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2006. / Submitted by denise santos (denise.santos@ufc.br) on 2014-02-19T13:42:07Z No. of bitstreams: 1 2006_dis_fasoares.pdf: 2441204 bytes, checksum: 165a38ea8594ecff8508c2b7a88eb873 (MD5) / Approved for entry into archive by denise santos(denise.santos@ufc.br) on 2014-02-19T13:42:47Z (GMT) No. of bitstreams: 1 2006_dis_fasoares.pdf: 2441204 bytes, checksum: 165a38ea8594ecff8508c2b7a88eb873 (MD5) / Made available in DSpace on 2014-02-19T13:42:47Z (GMT). No. of bitstreams: 1 2006_dis_fasoares.pdf: 2441204 bytes, checksum: 165a38ea8594ecff8508c2b7a88eb873 (MD5) Previous issue date: 2006 / The aim of this study is to analyse the frequence of anorectocele in adult women with obstructed defecation accordind to parity and age by means of cinedefaecography and anorectal eletromanometry. Forty-five adult women complaining of obstructed defecation were evaluated, with mean age of 46.3 years (23-72) and mean SCCC-C score of 13.3 points (6-23). Fifteen (33.3%) patients were nulliparous, seven (15,6%) primiparous and 23 (51,1%) multiparous, with mean parity per patient of 2.8 (0-11), considering only vaginal deliveries. Eighteen (60%) had a history of episiotomy, fourteen (46,7%) delivered macrossomic children and two (6,7%) had history of forceps-assisted delivery. Anal hipertony was verified in fourteen (31,1%) patients, while anal hipotony was present in eight (17,8%). Anismus was identified in thirteen (28,9%) patients. Anorecoceles were demonstrated in 34 (75,6%) patients, with mean size (TAR) of 24,8 mm (0-64). Thirty-six (80%) patients presented excessive perineal descent (DPM), rectal mucosal prolapse (PM) in 17 (37,8%) and rectoanal intussusception (IRA) in twelve (26,7%). There were no correlations between anorectocele and anal hipertony (p = 0,7171), anismus (p = 0,4666), IRA (p= 0,6991), PM (p = 0,2279), parity comparing nulliparous and multiparous patients (p = 1,000), episiotomy (p = 1,0000), forceps assistance (p = 1,0000) and delivery of macrossomic children (p = 1,0000). There were also no correlation between TAR and PMR (p =0,0883), PVM (p = 0,7327), parity (p = 0,4987) or age (p = 0,8603). There were correlations between anorectocele and DPM (p = 0,0275), score of SCCC-C (p =0,0082) and anal hipotony (p = 0,0141). In conclusion, anorectocele frequence is high and doesn’t correlate to parity, age but correlates to anal hipotony, DPM andconstipation. / O objetivo é avaliar a freqüência e o tamanho de anorretocele em mulheres adultas com evacuação obstruída, correlacionando-os com paridade, idade e parâmetros clínicos, utilizando cinedefecografia e eletromanometria. Foram avaliadas 45 mulheres adultas, com idade média de 46,3 anos (23-73) e sintomas de evacuação obstruída, com escore médio de 13,3 (6-23) pontos, segundo o Sistema de Classificação da Cleveland Clinic para Constipação (SCCC-C). Os parâmetros avaliados foram idade, dados obstétricos, escore do, SCCC-C, dados manométricos e achados de cinedefecografia.. Quinze (33,3%) pacientes eram nulíparas, 7 (15,6%) primíparas e 23 (51,1%) multíparas, com média de 2,8 (0-11) partos vaginais por paciente. Dezoito (60,0%) pacientes haviam sido submetidas a parto vaginal com episiotomia, sendo verificado feto macrossômico em 14 (46,7%) e aplicação de fórcipe em duas (6,7%) . Foi observada hipertonia esfincteriana em 14 (31,1%) e hipotonia em 8 (17,8%) pacientes. Foi identificado anismus em 13 (28,9%) pacientes. Foram demonstradas anorretoceles em 34 (75,6%) pacientes com tamanho (TAR) médio de 24,8 mm (0 - 64). Foram verificados descenso perineal móvel acentuado (DPM) em 36 (80%) pacientes, prolapso mucoso (PM) em 17 (37,8%) e intussuscepção reto-anal (IRA) em doze (26,7%). Não houve correlação entre anorretocele e hipertonia esfincteriana (p = 0,7171), anismus (p = 0,4666), IRA (p = 0,6991), PM (p= 0,2279), paridade comparando-se nulíparas e multíparas (p =1,000), episiotomia (p = 1,0000), uso de fórcipe (p = 1,0000), parto de feto macrossômico (p = 1,0000). Não houve correlação entre TAR e PMR (p = 0,0883), PVM (p = 0,7327), paridade (p = 0,4987) e idade (p = 0,8603). Houve correlação entre anorretocele e DPM (p = 0,0275), escore de constipação do SCCC-C (p = 0,0082) e hipotonia esfincteriana (p = 0,0141). Conclui-se que a freqüência de anorretocele foi elevada, não se correlacionou com paridade e idade, associando-se com hipotonia esfincteriana, DPM e constipação.

Faculty of Health Scicence School of Medicine 0100107g johannes.geyer@wits.ac.za / The 14C and 13C labelled urea breath tests (UBT) for detecting Helico-bacter pylori infection are well established but scope for improvement exists in both to reduce some of their shortcomings. For this study, the 14C UBT investigation focussed on reducing the quantity of radioactive tracer that is administered to the subject un-dergoing this test, with the aim of lowering the radiation dose to the patient, reducing the impact to the environment and exempting the test from radioactive materials licensing. Wider acceptance, availabil-ity, affordability to lower socio-economic groups and third party medi-cal treatment payers and using readily available equipment were fac-tors considered when developing the method. The principle of the method developed is to collect larger volume breath sample, quantitatively absorbing a defined volume of extracted breath CO2 in an efficient CO2 trapping agent using a specifically de-signed apparatus and measuring the activity with a low background β-spectrometer. A reduction in the quantity of 14C labelled urea administered to the pa-tient was achieved. The method also reduced the counting error mar-gin at a lower detection limit, improving discrimination between H. py-lori positive and negative patients. iii The 13C UBT is a non-radioactive test however, it is substantially more expensive. The 13C UBT investigation aimed to determine whether commercially available un-enriched urea could be used thus reducing the cost of the 13C UBT. A simple protocol with Isotope Ratio Mass Spectrometry (IRMS) for the measurement was used as opposed to the well-established 13C UBT protocol. The principle of the 13C UBT investigation was to detect the change of the breath δ13C (13C/12C) ratio after the administration of un-enriched urea with a δ13C different to the exhaled breath. Theoretical calculations showed that an administered dose of 500mg un-enriched urea with at least a 10‰ δ13C difference may be detectable using IRMS. In vitro investigations confirmed that levels of 0.01 to 0.001‰ δ13C were detectable by IRMS. A change in the δ13C of a standard breath CO2 was confirmed for a range between 0.14 to 50% v/v mixed CO2 samples, i.e. the projected range for in-vivo investigation. Results from the in-vivo investigation however were not able to distinguish positive from negative H. pylori patients. The use of the 1000mg dose of urea appears to have caused saturation of the enzyme. It was con-cluded that some enrichment of the 13C is necessary or less urea be used.

Un nombre grandissant d’associations entre diverses pathologies humaines et dysbiose intestinale (définie ici comme altération de la composition du microbiote par rapport à sa composition habituelle) sont observées. Parmi les facteurs qui pourraient induire la dysbiose, les bactériophages (dit phages), sont des candidats pertinents par leur fonction prédatrice.L’objectif de la thèse a été de déterminer si les prophages de souches bactériennes du microbiote intestinal humain ont un impact négatif sur la stabilité de leur hôte dans l’intestin. Pour cela, nous avons utilisé des souris sans germes primo-colonisées avec la souche Escherichia coli LF82, puis inoculées soit avec Faecalibacterium prausntizii A2-165, soit avec Roseburia intestinalis L1-82, deux souches appartenant aux espèces dominantes du microbiote intestinal humain. Chacune de ces souches possède deux prophages dans son génome, Lagaffe et Mushu pour F. prausnitzii, Jekyll et Shimadzu pour R. intestinalis. L’impact des prophages a également été étudié lors d’une inflammation intestinale induite au DSS.Pour la combinaison F. prausnitzii/E. coli, aucun des deux prophages de F. prausnitzii n’a d’activité délétère pour son hôte bactérien chez la souris, même durant une inflammation induite au DSS. Afin de mieux caractériser l’ensemble des prophages présents chez cette espèce, une analyse bio-informatique effectuée sur 15 souches de F. prausnitzii a permis de constater que la prévalence de Mushu et Lagaffe était faible, mais aussi de découvrir une remarquable richesse phagique : au total, 18 espèces de prophages répartis en nouveaux 8 genres viraux ont été décrits. Une étude in silico de l’abondance de ces phages dans les viromes intestinaux humains a révélé que des phages du genre ‘Lugh’ et ‘Epona’ sont plus souvent présents et/ou abondants dans les viromes de patients des maladies inflammatoires chroniques de l’intestin (MICI) que chez les individus sains. Sachant que les patients atteints de MICI ont une population appauvrie de F. prausnitzii dans leur microbiote, ces observations suggèrent une activité accrue de ces phages pendant la maladie : ils pourraient déclencher ou aggraver la baisse de population de F. prausnitzii dans les patients, participant ainsi à l’aggravation des symptômes des MICI.Avec la combinaison R. intestinalis/E. coli., aucune variation de population ou effet délétère du phage Jekyll n’a pu être observé. En revanche, la population du phage Shimadzu est loin d’être stable. Dans toutes les souris, et même en l’absence d’un traitement au DSS, un mutant virulent de Shimadzu émerge, appelé Shi-vir. Ce mutant lyse massivement la population intestinale de R. intestinalis, menant à un effondrement de la population hôte. La population bactérienne remonte ensuite à son niveau initial grâce à l’émergence de mutants bactériens résistants à l’infection. Cette résistance a essentiellement pour origine l’acquisition d’un espaceur associé au système CRISPR-Cas de type IIC de R. intestinalis, et dirigé contre le phage Shimadzu. Cependant, l’acquisition de cet espaceur ne peut se faire sans qu’une sous-population de R. intestinalis soit préalablement guérie du prophage Shimadzu, sans quoi un tel espaceur tuerait la bactérie.J’ai ainsi démontré qu’un prophage peut déstabiliser sa population hôte dans l’environnement intestinal et créer des dysbioses intestinales transitoires. La pression de sélection qui résulte de l’infection par le phage Shi-vir a permis l’accélération de l’évolution de l’hôte bactérien.En conclusion, une fraction des phages tempérés du microbiote intestinal pourrait avoir un impact négatif sur la stabilité de sa population hôte dans l’environnement intestinal, soit parce le ratio phage/bactérie augmente dans cet environnement (cas des phages Lugh et Epona de F. prausnitzii), soit parce qu’il évolue vers la virulence (cas de Shi-vir chez R. intestinalis) et induit une dysbiose transitoire. / A growing number of associations is observed between various human pathologies and intestinal dysbiosis, here defined as an alteration of the microbiota composition. Among the potential factors inducing dysbiosis, bacteriophages, called phages, are relevant candidates by their predatory function.The aim of the thesis was to determine whether prophages of bacterial strains from the human gut microbiota have a negative impact on the stability of their host in the gut environment. We studied this question by using germ-free mice colonized first with Escherichia coli strain LF82, then inoculated with two bacterial strains belonging to dominant species of the human intestinal microbiota, Faecalibacterium prausnitzii strain A2-165 or Roseburia intestinalis strain L1-82. Each of these strains has two prophages in its genome, Lagaffe and Mushu for F. prausnitzii, Jekyll and Shimadzu for R. intestinalis. The impact of these prophages was also studied during intestinal inflammation using DSS (Dextran Sulfate Sodium)-induced colitis in mice.In mice colonized with F. prausnitzii and E. coli , prophages of F. prausnitzii did not have any deleterious activity for the bacterial host, even during DSS-induced inflammation. In order to better characterize prophages of the F. prausnitzii species, a bioinformatic analysis carried out on 15 strains of F. prausnitzii highlighted that the prevalence of Mushu and Lagaffe was low. However, this analysis revealed also an enormous diversity of phages and we described 18 species of prophages divided into 8 new proposed genera. An in silico study of their abundance in 173 human intestinal viromes revealed that the phage genera 'Lugh' and 'Epona' were more present and/or abundant in viromes of Inflammatory Bowel Disease (IBD) patients compared to healthy subjects. Given that IBD patients have lower populations of F. prausnitzii in their microbiota compared to healthy subjects, our observations suggest an increased activity of these phages during disease. They may trigger or worsen population decline of F. prausnitzii in patients, participating thus to the aggravation of IBD symptomsIn mice colonized with R. intestinalis and E. coli, we did not observe variation of Jekyll population or deleterious effect of this phage on its host. In contrast, the Shimadzu population was not stable. Indeed, even in the absence of DSS treatment we observed in all mice the emergence of a virulent mutant of Shimadzu, called Shi-vir. This mutant massively lysed R. intestinalis, leading to a collapse of the bacterial host population. Then this population rose back to its original level thanks to the emergence of bacterial mutants resistant to the viral infection. This resistance was mainly due to the acquisition of a spacer associated with the CRISPR-Cas type IIC system of R. intestinalis, directed against the Shimadzu phage. However, acquisition of this spacer could not be observed unless the Shimadzu prophage was cured from the strain, showing that this spacer would kill the Shimadzu lysogen.I have shown therefore that a prophage can destabilize its host population in the intestinal environment and create transient intestinal dysbiosis. I have also highlighted that the selection pressure imposed by an ex-temperate phage infection, the Shi-vir phage, has allowed an acceleration of its host evolution.Overall, this work establishes that a fraction of the temperate phages present in intestinal microbiota may impact negatively bacterial population stability, either because the phage/bacteria ratio increases (for the Lugh and Epona phages de F. prausnitzii), or because the temperate phage evolves towards virulence (case of the Shi-vir mutant on R. intestinalis), and induces a transient dysbiosis.