Diss. (sammanfattning) Uppsala : Univ., 2007. / Härtill 4 uppsatser.

Orientadores: Silvana Pereira Barros, Sergio Roberto Peres Line / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-04T03:46:37Z (GMT). No. of bitstreams: 1 Vasconcelos_DanielFernandoPereira_M.pdf: 718453 bytes, checksum: 33e7ea7787a27dcce6c7a1cac2b7b5ed (MD5) Previous issue date: 2005 / Resumo: A doença periodontal (DP) é causada por interações entre fatores do hospedeiro, microrganismos específicos patogênicos e o sistema imunológico. TNF-b é um imunoregulador multifuncional que está relacionado com a patogênese de diversas desordens imunológicas, incluindo a DP. Nosso estudo analisou a associação entre DP e polimorfismo no gene TNF-b (+252 A/G). O DNA foi extraído de células da mucosa oral de 126 indivíduos brancos: 44 indivíduos controle e 82 indivíduos com DP. O polimorfismo foi analisado pela técnica de PCR, seguida pela RFLP. Os dados foram estatisticamente analisados pelo teste Exato Fisher (p<0,05) e Odds Ratio (OR). A freqüência do polimorfismo mostrou diferença estatisticamente significativa entre os grupos controle e com DP, revelando que indivíduos portadores do alelo G apresentavam 2,6 vezes mais chances de desenvolver a DP do que indivíduos saudáveis (G vs. A, p=0.0019, OR= 2.67, 95% CI 1.45 - 4.78), em relação aos genótipos a presença de pelo menos um alelo G predispõe 3,1 vezes à DP (G/G+ G/A vs. A/A, p=0,0059, OR= 3.1, 95% CI 1.45 - 6.65). Conclui-se que o TNF-b está envolvido na patogênese da periodontite crônica e pode ser utilizado como um marcador de risco para a DP na população estudada / Abstract: Background: Periodontitis is an inflammatory disease that leads to irreversible attachment loss, bone destruction and eventually bone loss, such cascade that culminates in tissue destruction initiates with pathogenic micro-organisms and depends on host response to disease expression. Tumor necrosis factor (TNF) a potent multifunctional immune modulator has been implicated in the pathogenesis of periodontal disease. Objective: In this study we investigated the hypothesis of association between chronic periodontitis (CP) and polymorphisms of the TNF-ß gene. Materials and Methods: One hundred twenty six individuals were evaluated by measuring clinical attachment loss and divided in 44 health individuals (control group-CG) and 82 subjects with CP. DNA samples were obtained from the individual's epithelial cells through scraping of the buccal mucosa. Polymorphism in the TNF-ß gene was analyzed by PCR, followed by NcoI restriction endonuclease digestion (RFLP). Results: The TNF-ß (+252A/G) polymorphism showed association with chronic periodontitis. Significant differences were found for the TNF-ß allele or carriage rate frequencies; odds ratio (OR)=2.67. Conclusions: These findings suggest that genotype composed of TNF- ß gene polymorphism may influence the susceptibility to chronic periodontitis / Mestrado / Histologia e Embriologia / Mestre em Biologia Buco-Dental

Hos vuxna individer förekommer två fenotyper gällande produktionen av laktas, vilka kallas laktostolerans och laktosintolerans. Vid laktosintolerans produceras otillräckliga mängder laktas vilket framkallar symptom som magsmärtor och flatulens vid intagandet av mjölkprodukter. En enbaspolymorfism (LCT-13910C&gt;T) har kopplats till laktostolerans hos nordvästeuropéer och kan genotypas med smältkurveanalys i realtids-PCR. På Laboratoriemedicin vid Länssjukhuset Ryhov används idag en metod vid genotypning av LCT-13910C&gt;T där extraktion av DNA från blod krävs innan analys. Anledningen till detta är att DNA-polymeraset som ingår enzymmixen LightCycler® FastStart DNA Master HybProbe endast fungerar med rent DNA-templat. Med en annan enzymmix, Kapa Probe Force, ska analys kunna göras direkt på blod. För att utvärdera enzymmixen jämfördes resultat från befintlig metod och resultat från metod med Kapa Probe Force, gällande förmågan att identifiera genotyperna LCT-13910C/C, C/T och T/T samt med avseende på imprecision. Vid jämförelse mellan metoderna samstämde resultatet i avseende på genotyp till 100 % utifrån specificerade smälttemperaturer (Tm) för respektive genotyp angivna i kitet för primer/prober. Däremot syntes lägre fluorescensnivå på smältopparna i metod med Kapa Probe Force, men påverkade inte tolkning av smältkurvorna. En lägre prov-till-prov-variation sågs även i resultatet från metod med Kapa Probe Force gentemot befintlig metod. / Among adults two phenotypes are found with regards to production of lactase, these are termed lactase persistence and lactose intolerance. Lactose intolerance is characterized by a low production of lactase, which leads to symptoms such as stomach ache and flatulence after the consumption of dairy products. A single nucleotide polymorphism (LCT-13910C&gt;T) has been correlated with the occurrence of lactase persistence in northwestern Europeans. Genotyping of LCT-13910C&gt;T is possible with melting curve analysis in real time PCR. The currently used method for genotyping of LCT-13910C&gt;T at Ryhov County Hospital requires the extraction of DNA template from blood, due to the fact that the DNA-polymerase in the kit LightCycler® FastStart DNA Master HybProbe requires pure DNA template for analysis. With another DNA-polymerase, included in the kit Kapa Probe Force, analysis on crude samples such as pure blood should be possible. Evaluation of Kapa Probe Force included comparison of the results from both methods with regards to identification of genotypes LCT-13910C/C, C/T and T/T and with regard to imprecision. The results from Kapa Probe Force were 100 % consistent with the results from existing method and acquired melting temperatures (Tm) were all within the accepted ranges specified in the kit of primers and probes. The fluorescence of melting curves acquired with Kapa Probe Force was significantly lower, however this had no effect when it came to interpreting the results. A lower variation could also be seen between samples with Kapa Probe Force compared to existing method.

Cada subtipo de HIV-1 tem um padrão mutacional próprio. Dados sobre mutações de resistência aos antiretrovirais foram obtidos com o subtipo B, primeiro em prevalência no Brasil. O segundo em algumas regiões é o subtipo F. Foram analisados padrões mutacionais em seqüências brasileiras de protease do subtipo F e levantou as seqüências deste subtipo disponíveis na base de dados de Stanford. A análise de dois grupos de seqüências (pacientes não tratados e tratados com inibidores de protease) mostrou 19 mutações associadas ao tratamento comuns ao subtipo B e 17 duplas de mutações associadas ao tratamento que diferem das descritas para o subtipo B, indicando a necessidade de estudos sobre rotas mutacionais no subtipo F. / Each HIV-1 subtype has a specific mutation pattern. Data on HIV-1 antiretroviral resistance mutations were obtained with subtype B, the first in prevalence in Brazil. The second in some regions is subtype F. Mutation patterns of Brazilian subtype F protease sequences were analyzed and performed a research of the sequences of Stanford Database. The analysis of two groups of sequences (untreated and treated patients with protease inhibitors) showed 19 treatment associated mutations also common in subtype B and 17 combinations of statistically treatment associated mutations that were quite different to those described for subtype B, indicating the need of studies to evaluate specific mutation pathways of subtype F.

O objetivo do trabalho foi de investigar os mecanismos genéticos relacionados à resposta humana à malária em duas populações Amazônicas. Cerca de 180 indivíduos de Portuchuelo e mais de 800 indivíduos de Monte Negro foram incluídos na amostra. Os indivíduos amostrados passaram por um levantamento epidemiológico e caracterização de marcadores genéticos. O número de episódios de malárias relatado pelos indivíduos, após correção para idade e sexo, foi submetido a testes de associação. A associação esperada entre indivíduos Fy- e a característica estudada foi encontrada nas duas populações, sem heterogeneidade significante. Análises de segregação complexa indicaram a presença de um gene principal relacionado ao acometimento de malária. Utilizando 108 STRs foi realizada uma varredura genômica na população de Portuchuelo onde foi encontrado um pico de lod score, sugestivo de ligação no braço curto do cromossomo 4. Os resultados obtidos em Portuchuelo não foram confirmados em 9 STRs estudados na população de Monte Negro. / With the aim of investigate the genetic mechanisms related to the human response to malaria infection, two Amazonian populations were studied. About 180 individuals from Portuchuelo population and more than 800 individuals from Monte Negro were included in the sample. Those individuals where subjected to an epidemiological survey, followed by characterization of genetic markers. Among the characteristics surveyed, the number of malaria episodes reported by the subjects was tested for association with classic blood groups markers after correction for age and sex. The expected association with Fy- individuals was observed in both populations, without significant heterogeneity. Complex segregation analyses indicated the presence of a major gene related to the number of malaria episodes. A genomic scan using STRs markers was conducted at the Portuchuelo sample. The multipoint linkage scan showed a lod score suggestive of linkage at the short arm of chromosome 4. The results found at Portuchuelo were not confirmed at the Monte Negro sample using 9 STRs.

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2006. / Härtill 5 uppsatser.

Cada subtipo de HIV-1 tem um padrão mutacional próprio. Dados sobre mutações de resistência aos antiretrovirais foram obtidos com o subtipo B, primeiro em prevalência no Brasil. O segundo em algumas regiões é o subtipo F. Foram analisados padrões mutacionais em seqüências brasileiras de protease do subtipo F e levantou as seqüências deste subtipo disponíveis na base de dados de Stanford. A análise de dois grupos de seqüências (pacientes não tratados e tratados com inibidores de protease) mostrou 19 mutações associadas ao tratamento comuns ao subtipo B e 17 duplas de mutações associadas ao tratamento que diferem das descritas para o subtipo B, indicando a necessidade de estudos sobre rotas mutacionais no subtipo F. / Each HIV-1 subtype has a specific mutation pattern. Data on HIV-1 antiretroviral resistance mutations were obtained with subtype B, the first in prevalence in Brazil. The second in some regions is subtype F. Mutation patterns of Brazilian subtype F protease sequences were analyzed and performed a research of the sequences of Stanford Database. The analysis of two groups of sequences (untreated and treated patients with protease inhibitors) showed 19 treatment associated mutations also common in subtype B and 17 combinations of statistically treatment associated mutations that were quite different to those described for subtype B, indicating the need of studies to evaluate specific mutation pathways of subtype F.

O objetivo do trabalho foi de investigar os mecanismos genéticos relacionados à resposta humana à malária em duas populações Amazônicas. Cerca de 180 indivíduos de Portuchuelo e mais de 800 indivíduos de Monte Negro foram incluídos na amostra. Os indivíduos amostrados passaram por um levantamento epidemiológico e caracterização de marcadores genéticos. O número de episódios de malárias relatado pelos indivíduos, após correção para idade e sexo, foi submetido a testes de associação. A associação esperada entre indivíduos Fy- e a característica estudada foi encontrada nas duas populações, sem heterogeneidade significante. Análises de segregação complexa indicaram a presença de um gene principal relacionado ao acometimento de malária. Utilizando 108 STRs foi realizada uma varredura genômica na população de Portuchuelo onde foi encontrado um pico de lod score, sugestivo de ligação no braço curto do cromossomo 4. Os resultados obtidos em Portuchuelo não foram confirmados em 9 STRs estudados na população de Monte Negro. / With the aim of investigate the genetic mechanisms related to the human response to malaria infection, two Amazonian populations were studied. About 180 individuals from Portuchuelo population and more than 800 individuals from Monte Negro were included in the sample. Those individuals where subjected to an epidemiological survey, followed by characterization of genetic markers. Among the characteristics surveyed, the number of malaria episodes reported by the subjects was tested for association with classic blood groups markers after correction for age and sex. The expected association with Fy- individuals was observed in both populations, without significant heterogeneity. Complex segregation analyses indicated the presence of a major gene related to the number of malaria episodes. A genomic scan using STRs markers was conducted at the Portuchuelo sample. The multipoint linkage scan showed a lod score suggestive of linkage at the short arm of chromosome 4. The results found at Portuchuelo were not confirmed at the Monte Negro sample using 9 STRs.

Made available in DSpace on 2014-08-20T13:32:57Z (GMT). No. of bitstreams: 1 tese_augusto_schneider.pdf: 1335904 bytes, checksum: 383222cf3cae0eb874a534a96d7cb6a1 (MD5) Previous issue date: 2011-03-29 / Fertility in cattle is highly influenced by metabolic hormones. Of special interest for the reproductive axis are growth hormone (GH) and insulin-like growth factor (IGF-I). The aim of this thesis were 1) to characterize liver GHR and IGF-I expression in postpartum lactating beef cows and its relation to first ovulation; 2) to characterize GHR and IGF-I expression in granulosa and luteal cells; 3) to determine the effect of prepartum somatotropin on the postpartum resumption of ovarian activity; and 4) to determine the effect of the GHR AluI polymorphism on the fertility of Holstein cows. The expression of GHR and IGF-I mRNA in the liver of postpartum lactating beef cows was not different between ovulatory and non-ovulatory cows, as well as it not change from 0 to 40 days postpartum. These data indicated that beef cows did not go through the same pattern of dissociation of the GH/IGF-I as observed for dairy cattle. Regarding the characterization of GHR and IGF-I expression in luteal and granulosa cells, it was possible to observe that GHR, IGF-I and SOCS expression were clearly higher in luteal than granulosa cells, but not different between atretic and estrogen active follicles. Moreover, GHR and IGF-I expression were not correlated in granulosa and luteal cells. With these results, we choose to focus on strategies in dairy cattle and not based on local regulation of GHR/IGF-I. In this sense, the use of prepartum somatotropin was benefitial in antecipate the resumption of postpartum ovarian activity. Further, treated cows had increased milk production and decreased concentrations of NEFA in the early postpartum. Also, the strategy based on the genotyping of Holstein cows for the GHR AluI polymorphism indicated that cows carrying the AluI(-/-) genotype had lower milk production and shorter calving conception interval. Cows carrying at least one AluI(-) allele also had a shorter calving conception interval and less number of AI per conception. This way, it is clear the importance of the GH/IGF-I system to the reproductive and productive performance of postpartum dairy cows. More studies, with new strategies or combining different strategies can bring even more benefits to the production systems. / A fertilidade em bovinos é muito influenciada por hormônios ligados ao metabolismo. Neste sentido, de especial interesse ao eixo reprodutivo são o hormônio do crescimento (GH) e o fator de crescimento semelhante a insulina I (IGF-I). Assim o objetivos desta tese foram 1) caracterizar a expressão de GHR e IGF-I no tecido hepático de vacas de corte lactantes e sua relação com a primeira ovulação; 2) caracterizar a expressão de GHR e IGF-I em células luteais e da granulosa; 3) avaliar o efeito da somatotropina exógena pré-parto sobre o retorno a atividade ovariana pós-parto; e 4) avaliar o efeito o polimorfismo AluI no gene GHR sobre a fertilidade de vacas da raça Holandês. A expressão de GHR e IGF-I no tecido hepático de vacas de corte lactantes não foi diferente entre vacas que ovularam e não ovularam, bem como não variou sua expressão entre 0 e 40 dias pós-parto. Estes dados indicam que vacas de corte não sofrem a dissociação do eixo GH/IGF-I observada em gado leiteiro. Com relação a caracterização da expressão de GHR e IGF-I em células luteais e da granulosa, foi possível observar que a expressão de GHR, IGF-I e SOCS é claramente maior em células luteais, porém não difere entre folículos atrésicos ou estrogênio ativos. Além disso, o nível de GHR e IGF-I não foi correlacionado em células da granulosa ou luteais. Assim, partiu-se para o teste de estratégias em gado leiteiro e que não focassem na regulação da produção local de IGF-I. Neste sentido, o uso de somatotropina exógena no período pré-parto mostrou um benefício ao antecipar o momento da primeira ovulação pós-parto. Além do mais, as vacas tratadas aumentaram a produção de leite e reduziram o nível circulante de ácidos graxos não esterificados no período pós-parto recente. Por fim, a estratégia de genotipagem de vacas da raça Holandês para o polimorfismo GHR AluI indicou que vacas portadoras do genótipo AluI (-/-) tem menor produção leiteira e menor intervalo parto-concepção. Em geral, vacas que possuem ao menos um alelo AluI(-) apresentaram menor intervalo parto-concepção e menor número de IA por concepção. Portanto, fica clara a importância do sistema GH/IGF-I para o desempenho tanto produtivo como reprodutivo de vacas de leite pós-parto, sendo que mais estudos com novas estratégias, assim como combinando diferente estratégias, podem trazer ainda mais benefícios aos sistemas de produção.

Für die Identifizierung bzw. Differenzierung von Candida- Spezies und -Stämmen sowie für die Bestimmung der genetischen und epidemiologischen Verwandtschaft von Stämmen der gleichen Spezies wurde eine PCR-Fingerprint-Technik und eine RFLP-Analyse der amplifizierten ITS-Region angewandt. Das PCR-Fingerprinting amplifiziert anonyme Sequenzen in der chromosomalen DNA, die über das gesamte Genom verteilt sind. Die ITS-Region ist Bestandteil des ribosomalen Operons, welches in ca. 50-100 Kopien/Zelle vorhanden ist. 1.a. Beide molekularbiologischen Verfahren wurden zur Unterscheidung von routinemäßig schwer differenzierbaren klinischen Candida famata und Candida guilliermondii-Isolaten genutzt. Von insgesamt 37 fraglichen Stämmen konnten 31 als C. guilliermondii und 3 als C. famata identifiziert werden, die drei verbliebenen Stämme waren mit diesen Techniken nicht identifizierbar. Mit der Biochemotypie gelang nur die Zuordnung eines der 3 C. famata-Isolate sowie von 23 der 31 C. guilliermondii-Isolate. 14 Isolate wurden mit den konventionellen Methoden gar nicht oder falsch identifiziert. 1b. Mit dem PCR-Fingerprinting wurde auch die Spezieszugehörigkeit phänotypisch veränderter Candida albicans-Isolate überprüft. Alle atypischen Stämme, bei denen solche für C.albicans charakteristischen Merkmale wie die Bildung von Chlamydosporen, die Verwertung der Aminozucker Glukosamin und N-Acetylglukosamin sowie die Assimilation von 2-Ketogluconat und Xylose nicht ausgeprägt waren, wiesen die für C. albicans typischen Fingerprintmuster auf. Unsere Studie zeigte, daß die biochemische Typisierung an Grenzen stößt, wenn typische Stoffwechselreaktionen nicht nachgewiesen werden können. 2. Bei 6 verschiedenen C. albicans-Populationen aus Angola, Madagaskar, Deutschland und Portugal wurde die Variabilität phänotypischer und genotypischer Merkmale untersucht, wobei in diese Analyse auch atypische Stämme miteinbezogen wurden. Während die phänotypischen Eigenschaften, bis auf die der atypischen Stämme, kaum variierten, wurden für die insgesamt 212 C. albicans-Isolate 87 unterschiedliche PCR-Fingerprint-Genotypen nachgewiesen. Eine Analyse der Beziehungen zwischen den Fingerprint-Genotypen wurde mit der UPGMA-Distanz-Methode durchgeführt. 3. Weiterhin wurden Candida-Vaginalisolate von Patientinnen mit rezidivierenden Episoden von Candida-Vaginitis mit Stämmen verglichen, die aus anderen Körperregionen stammten bzw. bei ihren Partnern isoliert wurden. Es konnte gezeigt werden, daß Stammaustausche zwischen den Partnern vorkommen, ein Stamm ohne oder mit geringfügigen genotypischen Veränderungen trotz Therapie persistieren kann und daß eine Reinfektion auch durch einen neuen Stamm möglich ist. Das Problem der Erregeridentifizierung in der mykologischen Labordiagnostik ist sowohl klinisch als auch epidemiologisch relevant. Molekularbiologische Methoden sollen gut funktionierende konventionelle Methoden zur Erregeridentifizierung nicht ersetzen, können aber bei Problemfällen eine wertvolle Ergänzung für die mykologische Diagnostik vorzugsweise in fachständigen Referenzlaboratorien darstellen. / A PCR fingerprinting approach and a RFLP analysis of the amplified ITS region were used to differentiate Candida species and strains as well as to assess genetic and epidemiological relationships of strains belonging to the same species. The PCR fingerprinting amplifies anonymous DNA sequences sampled throughout the whole genome. The ITS region is part of the ribosomal operon which occurs in tandem arrays of nearly 50-100 copies in one cell. 1.a. Both methods were used to identify clinical isolates of C. famata and C. guilliermondii which were difficult to differentiate with routine methods. Out of 37 ambiguous isolates 31 could be identified as C. guilliermondii, 3 as C. famata and the other 3 were not identifiable. Biochemical typing (Api 32 C V.1) identified only 23 out of 31 C. guilliermondii and 1 out of 3 C. famata whereas 14 isolates were misidentified or not identified at all. 1.b. By using the PCR fingerprinting technique strains of C. albicans with altered phenotypes could be identified at species level. Atypical isolates which did not express those characteristics which are thought to be typical for C.albicans like the formation of clamydospores, the ability to metabolise the amino sugars glucosamine and N-acetylglucosamine as the sole carbon source or to assimilate 2-ketogluconate and xylose showed the PCR patterns typical for C.albicans. Our study revealed that biochemical and morphological methods of species identification are limited if some of the key reactions fail. 2. We investigated the variability of phenotypic and genotypic properties of 6 different C. albicans populations from different countries (Angola, Madagascar, Portugal and Germany) including atypical strains. Except for the atypical strains only very little phenotypical variation was observed. However, 87 different genotypes were found among the 212 strains. The relatedness of the fingerprint-genotypes were analysed by measuring genetic distances with the UPGMA method. 3. Vaginal isolates of Candida spp. obtained from patients with recurrent episodes of vaginitis were compared with isolates from different body locations of the women and from their male partners. It has been shown that strain exchanges between the partners occur, that the original strain with or without minor genotypic changes can persist despite of the therapy, but also that reinfection by a new strain is possible. The identification of an ethiological agent by the mycological diagnostic laboratory is of clinical and epidemiological importance. Molecular biological methods should not replace well established conventional methods but they can supplement the identification of fungal pathogens in specialised reference laboratories if diagnosis cannot be achieved easily by conventional diagnostic procedures.