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- Lund, David, 1994
(författare)
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Computational discovery of antibiotic resistance genes and their horizontal transfer
- 2022
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Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Antibiotic resistance is increasing among clinical infections and represents one of the most serious threats to public health. Pathogens often become resistant by acquiring mobile antibiotic resistance genes (ARGs) via horizontal gene transfer (HGT). To limit the spread of new ARGs, it is important that we identify emerging threats early, and that we improve our understanding of what drives the HGT of ARGs. The three papers encompassing this thesis aim to increase our knowledge about ARGs and their mobility. In paper I, computational screening of large genomic datasets was used to identify new resistance genes for macrolide antibiotics, and to clarify their evolution. A large diversity of new erm and mph genes was identified, including six new families of mobile ARGs carried by pathogens, that showed varied phylogenetic origins. Of the tested genes, 70% induced resistance in Escherichia coli . In paper II, we identified previously undiscovered mobile genes giving resistance to aminoglycoside antibiotics in pathogens, further demonstrating how computational methods can discover potential emerging ARGs. Close to one million bacterial genomes were screened for aac and aph genes, and the mobility of each predicted gene was evaluated. A total of 50 families of new mobile ARGs were identified in pathogens. When new ARGs were tested in E. coli . 86% were functional, with 39% giving clinical resistance. In paper III, the factors influencing the HGT of ARGs were investigated. Phylogenetic analysis was used to identify HGT events from a large set of ARGs. For each event, the genetic compatibility of the involved gene(s) and genomes, as well as the co-occurrence of donor and recipient in different environments, were computed and used as input to train random forest classifiers. The resulting models suggested that the most important factor for determining if a mobile ARG successfully undergoes horizontal transfer is the genetic compatibility between the gene and the recipient genome. The findings presented in this thesis increase our knowledge about new genes giving resistance to two important classes of antibiotics. Furthermore, the results provide new insights into the horizontal transfer of resistance genes.
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| 2. |
- Pfister, Anna, 1984-
(författare)
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Outcomes of Myosin 1C Gene Expression Depletion on Cancer-related Pathways, in Vitro and in Clinical Samples
- 2016
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Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
- The unconventional myosin IC has previously been suggested to be a haploinsufficient tumour suppressor. The mechanism for this action has hitherto been unknown, however, and hence we decided to attempt to elucidate the genes involved. The first study involved knock-down of MYO1C using siRNA technology followed by whole transcriptiome microarray analysis performed on samples taken at different time points post transfection. This revealed a cornucopia of differential expressions compared to the negative control, among them we found an early up-regulation of the PI3K/AKT pathway and the pathway for prostate cancer. Among the down regulated pathways we found endometrial-, colorectal cancer and small cell lung cancer as well as the cell cycle pathway which was a little counter intuitive to the hypothesis that MYO1C suppresses cancer. For the next study six different genes (CCND1, CCND2, CDKN2B, CDKN2C, MYC, RBL1) important for the transitions into S-phase of the cell cycle were therefore chosen for validation using qPCR. These six genes and MYO1C were analysed on both the original time series and a new biological replicate as well as a well stratified set of endometrial carcinoma samples. We were able to verify the significant down-regulation of CCND2 in both time series indicating that this is caused by the depletion of MYO1C. In the tumour samples we saw a negative correlation between the expression of MYO1C and FIGO grade corroborating results previously found by our group when looking at protein expression.
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| 3. |
- Verma, Deepti
(författare)
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Genetic variations in the NALP3 inflammasome: a susceptibility factor for inflammatory diseases
- 2009
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Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Innate immunity has received impressive attention in the past decade owing to the discovery of the Toll like receptors (TLRs) and the NOD-like receptors (NLRs). While the TLRs specialize in fighting microbes at the cell surface, the NLRs complement by detecting and responding to intracellular microbes. Recently, the non-microbe sensing NLR called inflammasomes, have been identified, which senses metabolic stress as well as certain pathogenic microbes and elicits host’s inflammatory response. The NLR, NALP3 (formerly known as cryopyrin) forms a large cytoplasmic complex called the ‘inflammasome’ when NALP3, activated by a stimuli, associates with the adaptor proteins ASC and CARD-8. This interaction leads to the activation of pro-inflammatory caspase-1 which subsequently results in the formation of Interleukin (IL)-1β and IL-18. Mutations in the gene encoding NALP3, termed NLRP3 can lead to its constitutive activation resulting in an uncontrolled production of IL-1β. These mutations have been implicated in hereditary inflammatory diseases, often grouped under cryopyrin associated periodic syndromes (CAPS).This thesis describes a patient with a long history of arthritis and antibiotic resistant fever, but without the typical symptoms of CAPS. The patient was found to be a heterozygous carrier of two common polymorphisms Q705K in NLRP3 and C10X in the CARD-8. Experimental studies showed elevated levels of caspase-1 and IL-1β in the patient, and a total clinical remission was achieved by IL-1β blockade. These two polymorphisms combined, were found to occur in approximately 4% of the control population, suggesting the possibility of a genetic predisposition for inflammation in these individuals. Therefore, a cohort of rheumatoid arthritis (RA) patients, where elevated IL-1β could be one of the reasons behind chronic inflammation, was investigated. We found that carrying the combined polymorphisms resulted in increased RA susceptibility and a more severe disease course. Hypothetically, this subgroup of patients might benefit from IL-1β blockade. Additional studies are warranted to elucidate the functional effects of the two polymorphisms and to determine whether they identify a subgroup of patients that could benefit from IL-1 targeted therapy. Given the structural similarity of NALP3 to other NALPs, the possibility of involvement of the alternative, homologous genes cannot be eliminated.
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