Master of Science

thesisAccurate genotyping of the human leukocyte antigens (HLA) are crucial for the success of hematopoietic stem cell and solid organ transplantation. Over the past 50 years, numerous methodologies have been used for HLA typing but technological limitations have prevented full interrogation of the HLA gene thus resulting in allelic ambiguity. In order to resolve these ambiguities, additional testing is often required, leading to increased expense and delay in reporting the genotyping results. Recently, advances in nucleic acid sequencing technologies, generally referred to as next-generation sequencing (NGS), have become available. HLA genotyping by NGS is poised to become the new gold standard for several reasons. First, the entire gene can be interrogated, as opposed to a few exons, thus enabling greater resolution of known polymorphisms outside of the T cell recognition site on the HLA molecule. Secondly, missing genetic sequences in the curated databases will become more complete thus improving the algorithms used for alignment to a reference sequence. Lastly, NGS will become the new gold standard because high-resolution genotyping can be achieved without the need for additional time and laboratory resources in order to resolve allelic ambiguities and meet typing requirements required by regulatory agencies. In this thesis I discuss my efforts to employ this new technology by developing an assay for genotyping of the HLA-A, B, C, DRB1, and DQB1 genes. In the new assay, 98% of all alleles typed correctly and unambiguously without the need for any secondary testing. This in-house method is later compared with two recently released commercial kits and 100% concordance was found between the three methods. Differences in workflow are compared and contrasted. In conclusion, we show that HLA genotyping by NGS produces more correct and unambiguous results than traditional Sanger sequencing without the need for reflex testing

By-products in surface and reclaimed water disinfected with various agents

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Heat-Seal Ability and Fold Cracking Resistance of Kaolin-Filled Styrene-Butadiene-Based Aqueous Dispersions for Paper-Based Packaging

Dispersion coatings are offered as alternative solutions to extrusion coating technology for paper-based packaging. In addition to providing barrier properties, waterborne dispersions may implement the processing and converting properties of coated substrates, which are of extreme interest for an effective transfer to the industry. In this work, styrene-butadiene-based aqueous dispersions were formulated considering different amounts of kaolin as pigment. The authors assessed the heat-seal ability, fold cracking resistance, and blocking tendency, comparing the results against commercial dispersion coating grades. Kaolin content dominated the sealing behavior of experimental formulations, changing the minimum heat-seal temperature from 80 °C to >140 °C for 0% and 60% kaolin solid content, respectively. On the contrary, commercial grades were mostly affected by temperature. Additionally, despite the low latex glass temperature (0 °C), experimental formulations generally showed little, if any, blocking. On the downside, increasing kaolin content eases fold cracking, showing a different magnitude according to fold direction and coat orientation yet achieving a higher moisture barrier compared to commercial grades for both folded and unfolded samples

Information and Communications Technologies (ICTs) for energy efficiency in buildings: Review and analysis of results from EU pilot projects

Information and Communications Technologies (ICTs) can play a potential role in improving the energy performance of buildings by the implementation of effective solutions that take advantage of the energy interactions between all the elements included in a building. A revision of the 105 pilots implemented or under implementation in 18 projects in the area of ICTs for energy efficiency in buildings located in 23 European countries, through 88 cities with different types of climates, buildings and technologies have been carried out through documentary and field analysis of the energy, economic and social project results. These results have been extrapolated to assess the potential energy savings which could be expected at the EU level by implementing the solutions proposed by the projects. By the implementation of the different ICT solutions, buildings have achieved more than 20% energy savings. Pilots have demonstrated that the effectiveness of the ICT solution does not depend directly on the different climates where the solutions are implemented, but on several factors, such as the level of motivation, perceived thermal comfort, quality of social interaction and communication and ICT support

Recommendations for Sustaining Sustainability Initiatives in City Government

The report analyzes four U.S. cities' financial conditions and then compares and contrasts their sustainability initiatives and financing mechanisms. Based on the analysis, the report provides recommendations for other green cities that may make their sustainability initiatives more financially sustainable in today's fiscal environment.The report was a service learning project of a MPA class, Resource Allocation and Control, taught by Dr. Alfred T. Ho at the School of Public Affairs and Administration, University of Kansas in spring, 2012

Detection and distribution of European stone fruit yellows (ESFY) in apricot cv. ‘Bergeron’ and epidemiological studies in the province of Trento (Italy)

The aim was to investigate the performance of ‘Bergeron’ on ‘Wavit’ in 4 experimental fields, in the province of Trento (Italy), where European stone fruit yellows (ESFY) caused by “Candidatus Phytoplasma prunorum” has been constantly spreading since 2000.This included visual inspections for typical symptoms (early bud-break during dormancy and premature leaf-roll) and a highly sensitive Real time-PCR (Rt-PCR) assay. 25 % of the propagation material was checked with this method and found to be healthy, before planting in 2005.The epidemiology of the disease was also studied by focusing on: the presence of the vector Cacopsylla pruni (Scopoli) on conifers, the detection of “Ca. P. prunorum” in psyllid eggs and the transmission efficiency at different stages. This was done by exposing apricot trees in 2 locations, during 2 periods from January to July, to the overlapping presence in the orchards of the re-immigrants and the new generation of C. pruni. The results obtained demonstrated that ‘Bergeron’ seems to be highly susceptible to ESFY: typical bud-break was rarely observed, but up to 20-30% of the plants showed premature leaf-roll, fruit deformation and dieback. C. pruni was caught only once on Picea abies during winter; “Ca. P. prunorum” was found in 4 egg samples from 2 locations and the preliminary results on the exposed trees confirmed that the re-immigrants could be the most efficient vectors at least on apricot.Keywords: Prunus armeniaca, cultivar ‘Bergeron’, Real time-PCR, “Candidatus Phytoplasma prunorum”,epidemiolog

Caratterizzazione dell'efficacia di insetticidi verso Drosophila suzukii su mirtillo

Allo scopo di valutare le sostanze attive più efficaci nel controllo di Drosophila suzukii e di individuarne le modalità di azione, sono stati effettuate nel 2012 indagini di campo, semicampo e laboratorio. Le sperimentazioni, condotte su mirtillo, hanno coinvolto 7 principi attivi appartenenti a diverse famiglie chimiche. I formulati a base di fosmet hanno fornito un buono/ottimo contenimento del fitofago, con attività adulticida, di riduzione della deposizione di uova e di sviluppo embrionale. Acetamiprid ha dimostrato scarsa tossicità verso gli adulti, una ridotta limitazione dell’ovodeposizione e una parziale attività larvale. Spinosad ha indotto una buona mortalità degli adulti e un’apprezzabile riduzione dell’ovodeposizione, ma scarso contenimento nello sviluppo pre-immaginale. Spinetoram ha manifestato la stessa modalità di azione di spinosad, interferendo maggiormente sullo sviluppo del parassita e risultando complessivamente più efficace di spinosad. L’efficacia biologica di cyantraliniprole è risultata paragonabile e talvolta superiore alle performance di fosmet, interferendo in modo significativo sia per attività adulticida sia sullo sviluppo delle uova deposte; chlorantraniliprole ha mostrato un buon effetto adulticida quando utilizzato alle dosi maggiori, e valori intermedi verso gli altri parametri esaminati. Indoxacarb ha evidenziato una limitata attività complessiva nei confronti di D. suzuki