Development and characterisation of recombinant LSDV-vectored dual vaccines against bovine leukaemia virus and lumpy skin disease virus

Bovine leukaemia virus (BLV) and lumpy skin disease virus (LSDV) are endemic to Africa and cause significant economic losses to the beef and dairy industries. Vaccines are the most cost-effective and efficient way to prevent infection and outbreaks. Currently, there is no commercially available vaccine against BLV. In contrast, there are several live attenuated vaccines against LSDV. A recombinant LSDV which could protect cattle against both LSDV and BLV would be of great benefit to the African continent. This Master’s degree project involved three objectives. Firstly, the genetic variabilities and phylogenetic relationships of eight South African BLV isolates with other BLV strains from different geographical regions worldwide with known genotypes were determined. The BLV full-length envelope (env) and gag genes were successfully sequenced from total DNA extracted from the blood of BLV-infected cattle from a single herd. The analyses indicated that the seven of the South African isolates characterised in this study belonged to genotype 4 and the eighth to genotype 1. Furthermore, amino acid substitutions in the BLV Env and Gag sequences unique to the South African isolates were identified. Secondly, the activity of five selected poxvirus promoters in cells infected with LSDV was assessed by the detection of transient expression of an enhanced green fluorescent protein (eGFP) reporter gene driven by the poxvirus promoters. The promoters tested were a modified early fowlpox virus promoter (pmFP), an early-late promoter of a 7.5 kilodalton polypeptide gene of vaccinia virus (VACV) (p7.5), a synthetic early-late promoter of VACV (pS), a modified early-late promoter of the H5 gene of VACV (pmH5) and a synthetic early-late optimised promoter of VACV (pLEO). The results showed that all the poxvirus promoters were functional in the LSDV-infected cells and the eGFP expression was stable over the 72-hour study period. Lastly, two LSDV-vectored dual vaccines containing BLV immunogen(s) were developed and characterised. The first recombinant LSDV-vectored vaccine contained the BLV Env and Gag immunogens and the second recombinant LSDV-vectored vaccine contained the BLV Env immunogen alone. The presence of the BLV env gene in the recombinant LSDV vaccine was confirmed by polymerase chain reaction (PCR) and the BLV env sequence was confirmed by Sanger sequencing. Furthermore, BLV Env and Gag protein expression were confirmed by immunofluorescent staining and Western blotting, respectively. Future work will involve further purification of the recombinant viruses, confirmation of the production of BLV Gag virus-like particles and the preparation of high titre stocks of the vaccines to test in cattle

SEISMIC FRAGILITY ASSESSMENT OF CODE-CONFORMING BUILDINGS IN ITALY

The current European building codes, espousing performance-based seismic design principles, require safety verifications of structures under different levels of earthquakes. Although design seismic actions are prescribed on a probabilistically uniform basis referring to specific earthquake return periods, an explicit control of structural reliability of new constructions has not been implemented into the current practice. To assess implicit seismic risk of new constructions in Italy, a large national research project, named Rischio Implicito Norme Tecniche per le Costruzioni (RINTC), was developed for the years 2015-2017. One of the main contributions of this thesis is to gather insights into seismic risk of Italian code-conforming structures from a large building data set of the project. From a seismic design point of view, seismic performance of the prototype buildings is discussed through SDoF approximation of the detailed structural models, maintaing a wide variety of structural types, configurations, and seismic hazard levels at the building site. The study presented herein describes that heterogeneity of seismic risk among different structural types and different sites is mainly ascribed to actual strength reduction factors and local seismicity above the design seismic action. The thesis also provides seismic fragility functions of the examined prototype buildings, which are estimated through state-of-the-art approaches in performance-based earthquake engineering. For all examined buildings, fragility curves are estimated in terms of spectral acceleration at the fundamental vibration period of the structure, that is structure-specific. For the sake of comparison of structural fragility across multiple buildings, PGA-fragility curves are also examined for some selected building cases. To this aim, a probabilistic framework for hazard-consistent intensity measure conversion is established. Furthermore, the effects of seismic sequences in long-term seismic risk assessment of structures are also addressed given that the current seismic design philosophy neglects damage accumulation in structures

Trimmed Density Ratio Estimation

Density ratio estimation is a vital tool in both machine learning and statistical community. However, due to the unbounded nature of density ratio, the estimation procedure can be vulnerable to corrupted data points, which often pushes the estimated ratio toward infinity. In this paper, we present a robust estimator which automatically identifies and trims outliers. The proposed estimator has a convex formulation, and the global optimum can be obtained via subgradient descent. We analyze the parameter estimation error of this estimator under high-dimensional settings. Experiments are conducted to verify the effectiveness of the estimator.Comment: Made minor revisions. Restructured the introductory section

Topography and deposits of terrace of Yokohama Town, Northeast Japan

Higher, middle and Lower Terraces (Quaternary) have been distinguished in Yokohama Town, Aomori Prefecture. The Higher Terraces are subdivided into three terraces, H1,H2 and H3. They are distributed in hills. H1 and H2 deposites are composed of gravels of fan deposits. H3 deposits are composed of gravels, sands and mud of alluvial plain deposits. The Middle Terraces are subdivided into three terraces, M1, M1' and M2, and occupy almost all terraces in Yokohama Town. These deposits are composed of beach (sand with gravel) and fan (gravel) deposits. The Lower Terraces are subdivided into two terraces, L1 and L2. The L1 Terrace is distributed in limited area along coast and thought to be composed of beach deposits. The L2 Terrace is distributed in limited area along river and composed of braided stream deposits

Decoupling algorithm for evaluating multiple beam damages in steel moment-resisting frames

Post-earthquake safety evaluation of steel moment-resisting frames mainly relies on the inspection of seismic damage to beam–column connections. Recently, in order to evaluate seismic damage of steel connections in a prompt and precise manner, a local damage evaluation method based on dynamic strain responses has been proposed and receives attention. In the evaluation method where strain responses are measured by piezoelectric strain sensors, a strain-based damage index has been developed for evaluating individual seismic beam damage in a steel frame. However, for a steel frame suffering multiple beam damages, the damage index deteriorates its performance in identifying small damages with the presence of neighboring severe damages because of the moment redistributions induced by larger damages. This paper presents a decoupling algorithm that removes the issue of damage interaction and improves the performance of the damage index. The decoupling algorithm was derived on the basis of damage-induced moment release and redistribution mechanism. The effectiveness of the decoupling algorithm was numerically and experimentally investigated using a nine-story steel frame model and a large scale five-story steel frame testbed that can simulate multiple fractures at beam ends

Studies of the Synthetic Inorganic Ion Exchanger. V. : The Separation of Zirconium-95 and Niobium-95 by Means of a Stannic Phosphate Cation Exchanger

With the aim of developing a method for the separation of ^Zr and ^Nb, various eluants were investigated, a sulfuric acid solution was found to be most suitable for this purpose. After the stannic phosphate exchanger column had been conditioned with 1N nitric acid, the sample solution was passed through it to adsorb ^Zr and ^Nb. By using a 2N sulfuric acid solution and a 3N sulfuric acid -0.01N hydrofluoric acid solution as eluants, ^Zr and ^Nb could be eluted respectively. The separation was not quantitative, however, because of the unfavorable tailing of the elution curves. On the basis of the above results, the possibility of the total radiochemical separation of a long-lived fission product and the mechanism of the adsorption of these ions were discussed

An Investigation into the Effectiveness of Instructional Prompts in Sewing Classes : Based on an Analysis of Students' Reports on Yukata Sewing

本研究では, 浴衣製作実習において教師が提示した各授業プロンプトの有効性を検討することを目的としている。12回の実習授業で, 浴衣製作活動における学びを促すために教師によって提示された授業プロンプトの意味を学習者がどのようにとらえたか, その中で何を学んだかを28名の大学生の記述から分析した。その結果, 多くの学生は教師が提示した授業プロンプトから教授意図を理解し, それらを主体的かつ詳細にとらえ直していた。学生が記述した学びの内容は, 各種縫製技能や身体構造と被服構成などの実質的な事柄の他, 段取りの工夫や危険の回避など生活に必要な力も含まれていた。また繰り返し作業と繰り返しのない難易度の高い作業のいずれにおいても, 自己の内面的な変化への気づきや素材および道具, 技能と自分との関係性をみつめる記述がみられた。The aim of this paper is to investigate the effectiveness of teachers' instructional prompts when students are doing yukata sewing. This was done by analyzing 28 students' descriptions of their interpretations of prompts and what they learned in each of 12 lessons. A lot of students understood the teacher's intention and interpreted the prompt individually, and in detail. They acquired not only practical sewing skills and knowledge of the materials used, but also abilities which are necessary for daily life, for instance, the procedure for sewing and how to avoid dangerous situations. In repeating dull work as well as in doing difficult independent work, a lot of students reported awareness of their internal transformation and showed insights into the relationship between what they learnt about sewing, and themselves

Rapid and highly sensitive analysis of chlorophylls and carotenoids from marine phytoplankton using ultra-high performance liquid chromatography (UHPLC) with the first derivative spectrum chromatogram (FDSC) technique

We developed a rapid and highly sensitive analytical method for chlorophylls and carotenoids derived from marine phytoplankton using ultra-high performance liquid chromatography (UHPLC). High-performance liquid chromatography (HPLC) has been widely used in phytoplankton pigment analysis since the 1980’s for estimating the abundance, composition, and photosynthetic physiology of natural algal assemblages or laboratory cultures. However, the run-time of the HPLC analyses is generally ca. 30 min or more, which is time-consuming for analysts. Our UHPLC technique enabled us to complete the separations of chlorophylls and carotenoids from marine phytoplankton within 7 min with similar resolution as conventional HPLC methods. The analytical method was tested on authentic pigment standards, marine phytoplankton cultures, and field samples that were collected from the central tropical and subarctic Pacific plus the neritic Bering Sea. Critical pigment pairs that generally co-eluted as a single peak were successively resolved by obtaining the first derivative spectrum chromatograms (FDSCs) with a photodiode array (PDA) detector based on differences in pigment absorption spectra, e.g., chlorophyll (Chl) c2 and Mg 2,4 divinyl (DV) pheoporphyrin a5 monomethyl ester (MgDVP), as well as DVChl b and Chl b. Because the maximum injection volume of UHPLC is generally lower than that of HPLC to minimize the unwanted broadening of chromatographic peaks, the detection sensitivity needed to be increased, especially for oligotrophic seawater samples with low pigment concentration. To overcome this sensitivity issue, a PDA detector equipped with an 85 mm path length capillary cell was used with a fluorescence detector. As a result, the limit of quantitation (LOQ) as determined by absorbance was of the order of 0.1 ng for chlorophylls and carotenoids. Furthermore, a bead-beating technique using N,N-dimethylformamide (DMF) and zirconia beads was used to minimize the volume of the organic solvent utilized for pigment extraction. Our UHPLC method can replace the conventional HPLC techniques, and allows us to yield high-throughput data of the chlorophylls and carotenoids derived from marine phytoplankton