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Investigating host genetics and the role of selection for increased resistance to bovine tuberculosis in dairy cattleRaphaka, Kethusegile January 2018 (has links)
The significant social and economic losses as a result of bovine tuberculosis (bTB) present a continuous challenge to cattle industries in the United Kingdom (UK) and worldwide. Furthermore, as a zoonotic disease, bTB may pose a threat to humans. The potential transmission of bTB in cattle, estimated by the basic reproductive ratio (R0) was found to range between 1.0 and 1.9 in previous studies. In the UK, there has been an overall increase in bTB incidence in the last two decades despite national control and eradication programmes spanning over five decades. Such programmes mainly consist of surveillance based on the administration of skin tests and culling of animals reacting positive to these tests. Animal mobility restrictions are implemented in this case. At the same time, several studies have demonstrated that there is significant host genetic variation in individual cattle susceptibility to bTB, making the disease amenable to improvement with genetic or genomic selection. In addition, genomic analyses enhance the understanding of genetic mechanisms underlying the disease and its dynamics. The overall aim of this PhD thesis was to address existing scientific research gaps on the genetics of bTB resistance in dairy cattle. The following specific objectives were set: 1) to identify genomic regions underlying susceptibility to bTB using novel trait definitions, 2) to quantify the impact of long-term genetic selection for increased resistance to bTB on disease prevalence and dynamics and 3) to determine the consequences of genetically selecting for increased resistance to bTB on other economically important traits in dairy cattle. Genome-wide association studies (GWAS), regional heritability mapping (RHM) and chromosomal association analyses were applied in order to identify genomic regions associated with bTB (objective 1). Phenotypes comprised de-regressed estimated breeding values of 804 Holstein-Friesian sires obtain from the UK national genetic evaluation for bTB. Phenotypes pertained to three bTB trait definitions: i) positive reactors to the skin test with positive post-mortem examination results (phenotype 1); ii) positive reactors to the skin test regardless of post-mortem examination results (phenotype 2) and iii) as in (ii) plus non-reactors and inconclusive reactors to the skin test with positive post-mortem examination results (phenotype 3). In all cases, non-reactors without a subsequent positive post-mortem were considered to be healthy animals with regards to bTB. Genotypes based on a 50K SNP DNA array were available and a total of 34,874 SNPs remained after quality control. The estimated polygenic heritability for susceptibility to bTB was 0.26, 0.37 and 0.34 for phenotypes 1, 2 and 3, respectively. GWAS identified a putative SNP on Bos taurus autosomes (BTA) 2 associated with phenotype 1, and another on BTA 23 associated with phenotype 2. Genomic regions encompassing these SNPs were found to harbour potentially relevant annotated genes. RHM confirmed the effect of these genomic regions and identified new regions on BTA 18 for phenotype 1 and BTA 3 for phenotypes 2 and 3. Heritabilities of the genomic regions ranged between 0.05 and 0.08 across the three phenotypes. Chromosome association analysis indicated a major role of BTA 23 on susceptibility to bTB. A stochastic genetic epidemiological model based on four main disease states, namely susceptible (S), exposed (E), infectious (I) and test-sensitive (T), was developed to address objective 2. Effects of selection for increased resistance to bTB were investigated in a closed, genetically heterogeneous simulated population whose structure reflected the UK national dairy herd. Disease dynamics reflected real bTB data from the UK national genetic evaluation. The proposed genetic epidemiological model was implemented to simulate breakdowns under both absence and presence of selection. Genetic selection was simulated over 20 generations in 50 replicates, while exploring various selection intensities reflecting selection of the 10, 25, 50, 70 and 100% (no selection scenario) most resistant sires. Results indicated that selection significantly reduced the average underlying susceptibility across generations. The risk of breakdown was reduced by half after 4 and 6 generations for high selection intensities (10 or 25% of sires selected) and after 9 and 15 generations for low selection intensities (50 or 70% of sires selected). The average percentage of secondary cases was reduced to less than 1% in 4 and 5 generations for high selection intensities, and in 7 and 11 generations for low selection intensities. The reduction in the number of secondary cases across generations could also be indicative of the possible impact of genetic selection on the basic reproductive ratio (R0) which is defined as the number of secondary cases that results from an infectious individual in a naive population. Genetic selection also reduced severity and duration of breakdowns across generations. Finally, with regards to objective 3, a stochastic simulation was used to investigate the long-term effects of selection for resistance to bTB on other economically important traits in the UK dairy selection programme. Selection was simulated in a genetically heterogeneous population across 10 generations in 50 replicates. Animal genetic values for bTB and other traits were simulated based on variance and genetic correlation estimates obtained from literature. Independent culling levels selection of sires was applied in every generation whereby selection was first based on increasing resistance to bTB, then improving either an overall index, milk fat yield (FY) or milk protein yield (PY). This mimics real life practices regarding the newly released national genetic evaluations for bTB resistance. The overall index comprised several traits of interest such as milk yield (MY), FY, PY, feet and legs (FL), mammary (MAM), milk somatic cell count (SCC), calving interval (CI), non-return to service at 56 days (NR56) and lifespan (LS). A fertility index (FI) consisting of CI and NR56 was also considered in the analyses. Regarding bTB, different levels of selection intensities were explored corresponding to selection of the 10, 25, 50, 70 and 100% (no selection) most resistant sires. Two levels of selection intensity on the overall index, FY or PY were considered corresponding to selecting the best 5 and 10% of sires that were left after first selecting for bTB resistance. Results indicated that selection for increased bTB resistance would generally not have far-reaching consequences on other important traits. As expected, susceptibility to bTB declined with time and increasing selection intensity. Trends for all production traits (MY, FY and PY) in the present study were affected by selection for increased bTB resistance because of their significant genetic correlations with bTB. However, body conformation traits (FL and MAM) were not affected by selection for increased bTB resistance due to zero correlation assumed between these traits and bTB in the present study. Selection on bTB hampered improvement of SCC but enhanced LS because it was correlated unfavourably with SCC but favourably with LS. In all selection scenarios, the overall index improved and was generally not affected by selection for bTB resistance. Similarly, the FI was not affected by selection on bTB in all cases. However, secondary selection on production traits only (FY or PY) led to a decline in FI. Results presented in this thesis add insight into the genetic architecture of bTB and offer a prediction of potential effects of genetic selection for increased resistance to bTB in dairy cattle. The genomic regions and candidate genes identified to be associated with susceptibility to bTB will assist to further elucidate pathways critical to cattle susceptibility to bTB. / Consistent with previous studies of other populations and trait definitions, results from genomic association analyses suggest that susceptibility of cattle to bTB is heritable and likely a polygenic trait, amenable to improvement by genetic and/or genomic selection. Embarking on routine selection for resistance to bTB will reduce future bTB prevalence and severity of breakdowns across selection generations, as manifested by results of this thesis. The results also highlight the importance of considering selection as a complementary strategy to existing interventions. This has the potential to accelerate control and ultimate eradication of bTB. This strategy could assist the UK to achieve the national goal of being officially bTB free by 2038. Furthermore, as indicated by results of this thesis, selection against bTB in the national breeding programme will not adversely affect other economically important traits. Assimilation of bTB into the overall index will better manage possible antagonistic correlations between bTB susceptibility and some of the other traits.
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Caractérisation du gène XBTBD6 codant pour une protéine à domaine BTB-POZ impliquée dans la neurogenèse chez le xénopeBury, Frédéric Jacques 19 May 2006 (has links)
A la suite d’un criblage in silico nous avons identifié un nouveau gène codant pour une protéine à domaine BTB-POZ, XBTBD6.
Nous avons déterminé que la protéine XBTBD6 est une protéine cytoplasmique. Dans les cellules Hela, CHO, U2OS et COS7 la protéine XBTBD6 est localisée dans des corpuscules cytoplasmiques, localisation similaire à celle des protéines XBTBD3, HBTBD1 et HBTBD2. Nous avons observé que la partie N-terminale de la protéine, contenant le domaine BTB-POZ, est localisée dans la cellule comme la protéine entière ; par contre la partie C-terminale est exclusivement nucléaire. De plus, nous avons observé que XBTBD6 est localisée de façon diffuse dans le cytoplasme des cellules Neuro2A, 9L et 518A2e. Nous avons montré que la protéine XBTBD6 homodimérise et hétérodimérise avec XBTBD3 et XBTBD2 et qu’elle interagit avec l’ubiquitine ligase E3 XCullin 3. L’ensemble de ces interactions nécessite la présence du domaine BTB-POZ. Ces données montrent que les protéines BTBD6, BTBD3, BTBD1 et BTBD2 possèdent des propriétés communes indiquant qu’elles appartiennent à un sous groupe de la famille des protéines à domaine BTB-POZ.
Le profil d’expression a été analysé par la technique de protection à la RNAse et par hybridation in situ. Les résultats montrent que ce gène est fortement exprimé dans le système nerveux adulte et embryonnaire. Des expériences de surexpression par micro-injection d’ARNm ont permis de placer le gène XBTBD6 dans la cascade d’activation des gènes proneuraux en aval de XNgnr-1, XNeuroD, Xath3 et Xebf3. Ces résultats montrent que XBTBD6 est un marqueur neuronal chez le xénope.
Au cours de l’étude de la fonction du gène XBTBD6, nous avons montré que la surexpression et la perte de fonction de ce gène dans l’embryon de xénope n’induit pas de variation du nombre de neurones dans la plaque neurale. Par contre nous avons observé que la surexpression du gène XBTBD6 dans des cellules Neuro2A en différentiation régule négativement la croissance des neurites.
Nous avons élaboré un modèle de fonctionnement biochimique hypothétique où la protéine XBTBD6 fonctionnerait comme protéine adaptatrice dans un complexe d’ubiquitination permettant l’ubiquitination d’une protéine cible. Nous avons recherché les partenaires potentiels de XBTBD6 en utilisant la technique du double hybride en levure mais sans y parvenir.
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Investigating the role of ectoderm neural cortex 1 in osteoblast differentiationLeah Worton Unknown Date (has links)
The need for anabolic therapies to increase bone formation in difficult orthopaedic circumstances and to treat osteoporosis is an area of intense research focus. There is a current interest in the Wnt signalling pathway as a target for such treatment, with accumulating evidence for a role of this pathway in bone formation. Ectoderm Neural Cortex 1 (ENC1) is a Wnt target gene, not previously studied in bone, which was observed in our laboratory to be up-regulated in an anabolic surgical model of bone formation. The involvement of ENC1 in the differentiation of neuronal and adipocytic cells has previously been reported; therefore, this thesis investigates the expression of ENC1 in cells of the bone and the role of ENC1 during osteoblast differentiation. ENC1 transcript expression was localised to osteoblastic, chondrocytic and osteocytic cells in sections of healing fracture callus and normal mouse bone by in situ hybridisation. The expression of ENC1 was confirmed in differentiating primary osteoblasts and in osteoblastic and osteosarcoma cell lines by quantitative real time PCR and western blotting. ENC1 exists as two protein isoforms of 67 and 57kD in size, which are translated from alternatively spliced ENC1 transcripts. Both isoforms of the protein were detected in differentiating cultures of the pre-osteoblast cell line MC3T3-E1. To address the function of ENC1 in osteoblast differentiation, shRNA knockdown of the endogenous transcript was undertaken in MG63 osteosarcoma cells and in the MC3T3-E1 pre-osteoblastic differentiation model. Stable expression of shRNA targeted to both ENC1 spliceforms resulted in reduced accumulation of alkaline phosphatase positive nodules and alkaline phosphatase transcripts in MG63 cell culture. This reduction was not seen with targeted knockdown of 67kD ENC1 alone. Stable tetracycline-inducible shRNA knockdown targeted to both 57 and 67kD ENC1 isoforms in MC3T3-E1 cells resulted in a significant reduction of Alizarin Red S stained mineralised nodules. When expression of 67kD ENC1 alone was reduced, however, a significant increase in MC3T3-E1 nodule formation was observed. This knockdown had no effect on the expression of early genes involved in osteoblast differentiation Runx2 and osterix, but changes in expression of alkaline phosphatase and osteocalcin mRNA mirrored nodule formation. ENC1 is a member of the BTB-Kelch family of proteins. Some members of this family have recently been found to act as substrate adaptors for the E3 ubiquitin ligase, binding to the cullin 3 component of the complex. These adaptor proteins function to bring a substrate protein within the vicinity of the E2 ubiquitin-conjugating enzyme, thus targeting it for ubiquitination and subsequent proteasomal degradation. The ability of ENC1 to interact with cullin 3 was investigated as a possible mechanism by which it may affect a role in osteoblast differentiation. Full length ENC1 showed robust binding to cullin 3 and weak binding was seen between the N-terminally truncated 57kD isoform and cullin 3. ENC1, therefore, may act as a substrate adaptor protein for the cullin 3 based E3 ubiquitin ligase. These data present ENC1 as a novel candidate protein involved in osteoblast differentiation, and suggest the possible involvement of this protein in proteasomal degradation of a substrate involved in osteoblast differentiation. The ENC1 isoforms and the associated functional pathways thus are possible future therapeutic targets to treat bone loss and enhance or accelerate fracture healing.
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Control de orden fraccionario pi en convertidores de potencia back-to-backCrespo Herrera, Tedy Alain January 2017 (has links)
Magíster en Ciencias de la Ingeniería, Mención Eléctrica / En esta Tesis se presenta el uso de un control fraccionario proporcional-integral (FOPI) para convertidores de potencia, el cual es comparado con un control de orden entero proporcional-integral (PI), utilizado dentro del algoritmo de control vectorial, aplicadas a generadores síncronos de imanes permanentes (GSIP), operando con una red a frecuencia constante, interconectados mediante dos convertidores de potencia en una topología Back-To-Back (BTB). La idea de esta estrategia es transformar corrientes y voltajes en un marco de referencia giratorio (dq), donde las corrientes controladas son constantes en estado estacionario y posteriormente transformar las salidas de los controladores (FOPI y PI) en marco de referencia fijo (αβ) entregadas a un algoritmo de Modulación de Espacio Vectorial (SVM) que se encarga de generar los ciclos de trabajo necesarios para la conmutación de los elementos semiconductores que conforman los convertidores. El control del convertidor del lado GSIP tiene como objetivo principal el control de velocidad de la maquina eléctrica, mientras que el convertidor en lado de la red es, mantener el nivel de voltaje en el enlace de corriente continua (Dc-Link) que interconecta ambos convertidores. El sistema de control es tratado en el contexto de recuperación de energía en un mineroducto, pensado en que el sistema de control pueda tener la capacidad de realizar el mismo trabajo de las estaciones de choque, encargadas de mantener la velocidad de la pulpa mineral que transporta, sin embargo con este tipo de sistemas propuesto se podría recuperar parte del potencial energético disponible en el sistema y, emplearla de alguna manera más eficiente, proponiendo en este trabajo que sea conectada a una red eléctrica, buscando de esta manera que el sistema propuesto pueda ser una alternativa que cumpla la función de las estaciones disipadoras y además la recuperación de energía, aprovechando las ventajas de los controladores de orden fraccionario. Cabe mencionar que en este documento solo trata lo correspondiente a la parte de control del sistema eléctrico.
El estudio de comparación se realiza a nivel de simulación mediante el uso de programas computacionales Matlab-Simulink y Plecs, para los sistemas de control y los sistemas eléctricos respectivamente. Para determinar los escenarios de simulación del sistema se toma en cuenta los posibles escenarios que pueda presentarse en el mineroducto, siendo estos, una prueba sin posibles cambios en el torque de entrada, dos más cuando sufre cambios de +15% y -15% de la entrada nominal, una más cuando la entrada del sistema se torna de forma oscilante entre ±15%, en la última de las simulaciones se realizan combinación de controladores FOPI y PI dentro del algoritmo de control.
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Study of the prevalence of bovine tuberculosis in Govuro District, Inhambane Province, MozambiqueMacucule, Baltazar Antonio 02 March 2010 (has links)
This study was conducted to confirm the presence of bovine tuberculosis (BTB) and determine its prevalence, based on skin test reactivity, in cattle reared under extensive farming conditions in the Govuro district, Inhambane province, Mozambique. The study was comprised of a primary screening test using the single intradermal test (SIT) in randomly selected animals from Colonato and Sede dip tanks in Govuro. Positive reactors to the SIT were tested again with bovine and avian tuberculin using the single intradermal comparative test (SICTT) 7 weeks after the SIT. The sample size was calculated using Win Episcope 2.0 based on 95% confidence to detect a 2% expected prevalence using the SIT, with a 1% accepted error and accounting for a total population size of 7208. The calculated sample size was 682 animals. To compensate for the probability of 20% default in reading, the sample size was increased to 853. During the testing process (SIT), it was evident from the first 3 reading days that the apparent prevalence (61, 94%) was higher than expected (2%), hence we decided to stop when the total number of cattle was 530. During the testing process (SIT), it was evident from the first 3 reading days that the apparent prevalence (61.94%) was far higher than expected (2%), hence we decided to stop when the total number of cattle was 530. This was due to the fact that, at such a high prevalence, it would not be necessary to achieve as high a precision as 1% accepted error. A sample size of 530 would be sufficient to achieve a precision of 4% accepted error, which was regarded as more than adequate. The 530 cattle, 3 or more years of age, were selected using systematic random sampling from the two dip tanks (Colonato 371 and Sede 159 animals). All animals were identified by numbers painted, dorsally on the sacral region. Out of 530 tested cattle by SIT, 268 were read, and 166/268 (61.94% with 95% confidence interval [CI]: 55.8 – 67.8%) were found positive, with visible swallow at the injection site. Apparent prevalence (AP) was found to be 61.94% while the true prevalence (TP) was 75.92%. The predictive value of a positive result (PV+) was found to be 87.9%. No significant difference in apparent prevalence between the two areas was detected by Fisher’s exact test (P = 0.11). By SICTT, out of 28 animals positive reactors to SIT, 21 were possible to read, and 13/21 (61.9%; 95% CI: 55.1 – 89.3%) were found positive. A three year old bull, positive reactor to the SIT, was slaughtered, and a detailed post mortem was carried out and organs with visible lesions were collected for further laboratory testing (histopathology, culture and isolation of M. Bovis and PCR). Later on, 30 more positive reactors to the SIT test were slaughtered: 25/30 (83.3%) showed visible lesions compatible with BTB, and total condemnation of carcass was made in 3/25 (12%) due to generalized lesions. The high prevalence rate of skin test positive animals as well as gross lesions and histopathology were confirmed to be BTB by the isolation and identification of M. Bovis by culture and PCR. Our results suggest that bovine tuberculosis is highly prevalent in Govuro district and may thus represent a potential health problem of zoonotic tuberculosis in humans. Our results suggest that BTB has reached the plateau phase of endemicity in cattle in Govuro district. In this context, the positive predictive value of the SIT is very high and thus the use of the SICTT as a confirmatory test has a limited value and should not be advocated. Our results further indicate that no other prevalence study of BTB should be conducted in the next few years in Govuro district, unless comprehensive control measures are implemented. The focus of further studies should be on the isolation and the molecular characterization of M. Bovis from cattle and humans in order to assess transmission routes and the role played by BTB in human TB cases in Govuro district. Copyright / Dissertation (MSc (Veterinary Tropical Diseases))--University of Pretoria, 2009. / Veterinary Tropical Diseases / unrestricted
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Understanding the genetic and morphological basis of bushy root and bifuricate, two mutations affecting plant architecture in Solanum lycopersicum LSilva Ferreira, Demetryus January 2017 (has links)
The classical ethyl methanesulfonate (EMS) tomato mutant bushy root (brt) was studied using a homozygous near isogenic line (brtNIL) in the Micro-Tom (MT) genetic background. The mutation has a pleiotropic phenotype comprising slow seedling development, which may be a consequence of a maternally-inherited small seed phenotype, and a more compact, smaller but not bushier, root phenotype. The number of lateral roots, total root length and taproot size are all smaller in brtNIL than the WT. The BRT locus was mapped to a 137 kbp region containing 9 candidate genes on chr 12; an InDel in the promoter region of Solyc12g014590 – containing two highly conserved pirin domains (Pirin_C and Pirin), was detected. Different expression patterns were confirmed by transcriptomic results, supporting Solyc12g014590 as the gene responsible for the brt phenotype. A naturally occurring recessive mutant named bifuricate (bif) shows an increase in inflorescence (truss) branching in comparison to the wild type (WT) control line, LAM183. In addition, the number of flowers per truss was 235% higher in bif plants than WT. Low temperature is known to increase truss branching, and so a four day low temperature treatment was applied and it was demonstrated that flowering increased significantly more in bif than in LAM183. The BIF locus was mapped to a 2.01 Mbp interval of chromosome 12 containing 53 genes. All coding region polymorphisms in the interval were surveyed, and two genes Solyc12g019420 (a BTB/TAZ transcription factor) and Solyc12g019460 (a MAP kinase) contained one stop codon predicted to disrupt gene function; both genes are excellent candidates for inflorescence branching control based on literature evidence. A newly developed introgression browser was used to demonstrate that the origin of the bif mutant haplotype is Solanum galapagense.
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Molecular mechanisms of PLK1 recognition by CUL3/KLHL22 E3-ubiquitin ligase controlling mitotic progression / Mécanismes moléculaires de reconnaissance de PLK1 par l’E3-ubiquitine ligase CUL3/KLHL22 contrôlant la progression mitotiqueMetzger, Thibaud 25 March 2014 (has links)
L’ubiquitination est une modification post-traductionnelle impliquée dans de nombreux mécanismes cellulaires. L’E3-ubiquitine ligase CULLIN 3 (CUL3) est un régulateur essentiel de la progression mitotique, ubiquitinant d’importants régulateurs mitotiques et contrôlant leur localisation subcellulaire. Plus particulièrement, notre travail décrit le rôle de la nouvelle E31 ligase CUL3/KLHL22 dans la régulation de l’activité localisée de Polo-like kinase 1 (PLK1) et de ce fait dans l’établissement d’une progression mitotique précise. Néanmoins, les mécanismes moléculaires qui régissent la reconnaissance de son substrat par CUL3 demeurent inconnus. L’activité catalytique de PLK1 ne semble pas être nécessaire à son interaction avec KLHL22, mais aussi bien son domaine kinase que Polo-box (PBD) suffisent à co-purifier KLHL22. Des mutations au niveau du motif DFG, situé en amont du domaine kinase,et du tryptophane 414 au sein du PBD semblent influer sur la reconnaissance de KLHL22. Les résultats obtenus montrent les premières indications biochimiques du mode d’interaction du complexe CUL3/KLHL22/PLK1. / Ubiquitination is a post-translational modification involved in many cellular processes. The E3 ubiquitin-ligase based on CULLIN 3 protein (CUL3) is an essential regulator of mitotic division in human cells by ubiquitinating several important mitotic regulators and controlling their subcellular localization. In particular, our work described the role of novel CUL3/KLHL22 E3-ligase in regulation of localized activity of Polo-like kinase 1 (PLK1) and there by faithful mitotic progression. However, the molecular mechanisms of substrate recognition by CUL3 remain unknown. The catalytic activity of PLK1 may not be required for binding KLHL22 but both the kinase and the Polo-box domains are sufficient to co-purify KLHL22. Mutating the DFG motif within the kinase domain and the tryptophan 414 within the PBD influence the binding to KLHL22. These results provide first insights into molecular mechanisms of CUL3/KLHL22/PLK1complex.
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The intra- and inter-population relatedness of bovine tuberculosis-infected and -uninfected African buffaloes (Syncerus caffer caffer) in the Kruger National ParkRossouw, Ingrid 21 June 2011 (has links)
The African buffalo (Syncerus caffer) is a member of one of Africa’s most well known tourist attractions and unique grouping of mammals – the ‘big five’. Historical records indicate that during the 19th century approximately 3 million African buffaloes inhabited almost the whole of sub-Saharan Africa. Several factors such as disease, habitat fragmentation, over-hunting and drought reduced the buffalo population to approximately 400 000 by 1990. The African buffalo is host to a variety of sub-acute diseases, such as bovine tuberculosis (BTB), foot-and-mouth disease (FMD) and corridor disease (CD). Disease is an important factor which influenced African buffalo populations throughout the continent and more specifically the Kruger National Park (KNP) and is largely responsible for the fact that buffaloes are restricted to enclosed areas with strict regulations imposed on their movement. The social organization of animals influences the distribution and spread of a disease - especially in the case of the African buffalo in the KNP. The emergence of BTB in the largest conservation area in South Africa (the KNP), threatens wild and domestic animals and humans who are in close proximity to the Park. The potential economic losses associated with this disease are excessive. The results presented in this thesis provide baseline information into the genetic status of sampled African buffaloes in the KNP, genetic relatedness between sampled individuals as well as BTB associations between sampled African buffaloes in the KNP, based on a limited dataset of 181 animals. Twelve microsatellite markers were used to evaluate 181 samples which were collected from 39 locations dispersed throughout the KNP. Specific population genetic parameters revealed information based on the intra and inter - relationships at the ‘per population’ level as well as at the ‘per prevalence group’ level. Evidence indicates a medium to high level of genetic diversity, a low to medium level of inbreeding (inbreeding coefficient (Fis) for each group ranges between 0.143 and 0.147) and a relatively high level of migration for buffaloes associated with each prevalence group. Pairwise relatedness estimates were determined between individuals, to reveal their level of relatedness (unrelated, full siblings, parent-offspring or half siblings), based on Queller and Goodnight’s (1989) coefficient of relatedness. Relatedness was determined on different levels, intra and interpopulation level, BTB infected and BTB uninfected group level as well as prevalence group levels. Evaluation of data based on these different levels and between different groups, painted an overall picture of the disease condition and genetic relatedness within and between sampled BTB infected and BTB uninfected buffaloes. Evidence indicated that the greater majority of our sampled African buffaloes (BTB infected or uninfected), were genetically unrelated (in terms of sibling and parent-offspring relationships), irrespective of their disease status. M. bovis infected buffaloes sampled and used in our study are not more closely related to each other than to uninfected buffaloes in the same population or prevalence group. / Dissertation (MSc)--University of Pretoria, 2010. / Production Animal Studies / unrestricted
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The effect of methamphetamine on the blood-testis barrierZabida, Omer Saleh January 2018 (has links)
>Magister Scientiae - MSc / Introduction
The blood-testis barrier (BTB) is formed by tight junctions between adjacent Sertoli cells. The barrier formed by these tight junction helps to create a specialized environment for spermatogenesis and provide an immunological barrier to protect developing germ cells. Methamphetamine (Meth) is known as neurotoxin however, its effects on the male reproductive system, especially on Sertoli cells and, the BTB are not well established. Therefore, this study aimed to determine the effects of Meth on the TM4 mouse testis Sertoli cell line and on the integrity of the BTB permeability.
Materials and Methods
This study investigated the effect of selected concentrations of Meth (0.1 μM, 1 μM, 10 μM, 20 μM and 100 μM) on TM4 mouse testis Sertoli cell line for 24 until 96 hours, using two treatments: an “acute” study (24 hrs exposure) and a “chronic” study, where treatment occurred on a daily basis over 96 hrs. The following parameters were investigated: viability, cell proliferation, mitochondrial activity, monolayer permeability.
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BTB Domain Dimerization:Development of a Protein-protein Interaction AssayWang, Qingniao 22 September 2009 (has links)
In the human genome, 43 BTB (Bric-à-brac, Tramtrack, and Broad Complex) containing BTB-Zinc Finger proteins have been identified, many of which are transcription factors involved in cancer and development. These BTB domains have been shown to form homodimers and heterodimers which raise DNA binding affinity and specificity for transcription factors.
This project was to develop an efficient assay to systematically identify interactions between BTB domains. It combined a co-expression system, fluorescent protein tagging and Ni-NTA plate retention. It was concluded that fourteen analyzed BTB domains formed homodimers, but only certain BTB pairs formed heterodimers, such as BCL6 with Miz1 and Miz1 with RP58. To further understand the specificity of BTB domain interactions, more structural and sequence information is still needed. In conclusion, this assay provided a comprehensive detection method for BTB domain interaction mapping. The information generated provides candidates for further functional and structural studies.
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