Les cellules ont développé plusieurs voies de signalisation et de réponses transcriptionnelles pour réguler leur taille et coordonner leur croissance et leurs divisions cellulaires. L'intérieur des cellules est naturellement surchargé par des macromolécules. Cet encombrement macromoléculaire, appelé crowding, a été intensément étudié in vitro et est connu pour affecter la cinétique des réactions. Cependant, l'étude des effets d'encombrement in vivo est plus difficile en raison du haut niveau de complexité et d'hétérogénéité à l'intérieur d'une cellule. Au cours de cette thèse, nous nous sommes intéressés aux effets de changement du volume cellulaire sur la cinétique de réactions biochimiques chez la levure Saccharomyces cerevisiae. Pour cela, nous avons induit des stress osmotiques pour comprimer la cellule et étudier l'impact du crowding sur les cinétiques de signalisation. La réduction du volume cellulaire augmente la viscosité interne et peut retarder le fonctionnement de plusieurs voies de signalisation et de processus cellulaires. En augmentant progressivement le niveau de compression, on observe un ralentissement des processus biologiques jusqu'à un point où l'adaptation cellulaire est abolie. Ceci a été observé pour la translocation nucléaire de facteurs de transcription (Hog1, Msn2, Crz1, Mig1 et Yap1) ainsi que pour la mobilité des protéines Abp1 et Sec7. Nous montrons aussi que la compression altère la capacité de plusieurs protéines à diffuser dans le cytoplasme de différents types cellulaires. Nous proposons que ces altérations cinétiques induites par l'augmentation de la viscosité intracellulaire ne soient pas sans rappeler une transition vitreuse. Ces résultats suggèrent l'importance d'un encombrement macromoléculaire optimal permettant aux cellules de fonctionner correctement.

Bone discolouration of pig carcasses is a quality concern that has been observed in Ontario slaughter plants. The objectives of this study were to establish the prevalence of pig carcasses showing bone discolouration, its relationship with residues of tetracyclines in bones, and to investigate the use of tetracyclines in feeding programs for grower-finisher pigs as the main risk factor for discolouration. Abattoir data were examined to determine the extent of the problem and the prevalence of bone discolouration during 2006, 2008, 2009, and 2010 was found to be 0.13%, 0.22%, 0.26%, and 0.28%, respectively, indicating that the issue of bone discolouration was present at low levels over the entire period of the study. A controlled trial using feed, water, and injectable tetracycline products to investigate the effect of tetracyclines on residue and bone colour was conducted. Bones were assessed visually for signs of discolouration, and high performance liquid chromatography (HPLC) was used to measure the levels of tetracycline residues in the bones. Results from this trial demonstrated that discolouration could be produced with 660ppm of chlortetracycline (CTC) in feed for 12 weeks even when 33 days of withdrawal time was observed. It was also found that residues of tetracyclines can be present in bones in the absence of discolouration. A retrospective study was conducted to investigate tetracycline use in herds identified as having discoloured bones at slaughter. Positive shipments were associated with dosage and duration of CTC use as well as with length of withdrawal. In conclusion, discoloured bones of pig carcasses were identified at low levels in one large Ontario abattoir; however, further investigation is needed in order to determine the impact it may have on the swine industry. / Ontario Pork and the University of Guelph - Ontario Ministry of Agriculture (OMAFRA) Sustainable Production System Program

Root-lesion nematodes, Pratylenchus spp., are serious pathogens of potato plants worldwide. Several management practices can control Pratylenchus spp.; however, they all have shown some limitations. Therefore, environmentally-safe, low-cost, and effective control strategies are needed as possible alternative to currently used strategies. This thesis was designed to assess if liquid hog manure (LHM) holds such potential. The objectives of this thesis were to determine: (i) the prevalence and identity of species of Pratylenchus spp. in Manitoba potato fields, (ii) if short-chain volatile fatty acids (VFA) in LHM are the constituents responsible for the manure’s toxicity to Pratylenchus spp. using solution exposure experiments (iii) the effectiveness of LHM in killing Pratylenchus spp. in soil, and (iv) the impact of LHM on nematode communities. Pratylenchus spp. were detected in 39% of 31 potato fields surveyed in Manitoba with population densities ranging, for positive fields, from 45 to 631 nematodes kg-1 fresh soil. Morphometrics of female nematodes and molecular diagnosis (using species-specific PCR primers) showed that the species of Pratylenchus present in the potato fields to be P. neglectus. Potato, cv. Russet Burbank, showed to be a poor host to two populations of Pratylenchus spp. from Manitoba potato fields. Accordingly, P. neglectus does not seem to be a limitation to potato production in Manitoba; thus, P. penetrans, the most widely spread and damaging species to potato was used in the successive studies of assessing the use of LHM to control Pratylenchus spp. in potato fields. VFA (acetic, propionic, n-butyric, isobutyric, n-valeric, isovaleric, and n-caproic acids) accounted for the majority of the lethal effect of LHM to P. penetrans under acidic conditions. VFA in LHM killed Pratylenchus spp. in soil and acidification seemed to enhance its ability when VFA concentration in the manure is low. LHM did not act as a soil fumigant eliminating soil trophic interactions but increased bottom-up food web interactions. VFA in LHM persisted in the soil for four days with biological degradation being their mode of loss. In conclusion, LHM is potentially an effective and low-cost strategy to control Pratylenchus spp. and its efficacy can be improved by acidification.

Root-lesion nematodes, Pratylenchus spp., are serious pathogens of potato plants worldwide. Several management practices can control Pratylenchus spp.; however, they all have shown some limitations. Therefore, environmentally-safe, low-cost, and effective control strategies are needed as possible alternative to currently used strategies. This thesis was designed to assess if liquid hog manure (LHM) holds such potential. The objectives of this thesis were to determine: (i) the prevalence and identity of species of Pratylenchus spp. in Manitoba potato fields, (ii) if short-chain volatile fatty acids (VFA) in LHM are the constituents responsible for the manure’s toxicity to Pratylenchus spp. using solution exposure experiments (iii) the effectiveness of LHM in killing Pratylenchus spp. in soil, and (iv) the impact of LHM on nematode communities. Pratylenchus spp. were detected in 39% of 31 potato fields surveyed in Manitoba with population densities ranging, for positive fields, from 45 to 631 nematodes kg-1 fresh soil. Morphometrics of female nematodes and molecular diagnosis (using species-specific PCR primers) showed that the species of Pratylenchus present in the potato fields to be P. neglectus. Potato, cv. Russet Burbank, showed to be a poor host to two populations of Pratylenchus spp. from Manitoba potato fields. Accordingly, P. neglectus does not seem to be a limitation to potato production in Manitoba; thus, P. penetrans, the most widely spread and damaging species to potato was used in the successive studies of assessing the use of LHM to control Pratylenchus spp. in potato fields. VFA (acetic, propionic, n-butyric, isobutyric, n-valeric, isovaleric, and n-caproic acids) accounted for the majority of the lethal effect of LHM to P. penetrans under acidic conditions. VFA in LHM killed Pratylenchus spp. in soil and acidification seemed to enhance its ability when VFA concentration in the manure is low. LHM did not act as a soil fumigant eliminating soil trophic interactions but increased bottom-up food web interactions. VFA in LHM persisted in the soil for four days with biological degradation being their mode of loss. In conclusion, LHM is potentially an effective and low-cost strategy to control Pratylenchus spp. and its efficacy can be improved by acidification.

Submitted by Katiane Souza (katyane.souza@gmail.com) on 2016-04-07T00:06:08Z No. of bitstreams: 1 arquivototal.pdf: 13382272 bytes, checksum: 7f87ecfccf2692fd7d07c6a65a464403 (MD5) / Made available in DSpace on 2016-04-07T00:06:08Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 13382272 bytes, checksum: 7f87ecfccf2692fd7d07c6a65a464403 (MD5) Previous issue date: 2007-03-02 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Because extractive exploration, irregular production and the lacking management proposal concerning to its phytotechnical aspects, the hog-plum plant (Spondias mombin L.) is under marginal condition in the productive chain. So, this study was conducted to identify and quantify the nutritional demands of the hog-plum plant (Spondias mombin L.) for both phosphorus and potassium. The experiment was carried out in the experimental station of the Pesquisa Agropecuária da Paraíba - EMEPA-PB, Mangabeira county. The randomized block experimental desing was used, and the treatments were disposed under the factorial scheme 4 x 4 + 2, with the first two factors consisting of the P and K doses (30, 60, 120 and 240 g plant-1), and two controls (without fertilization and with organic fertilization). In estimating the yield, only the fruits fallen on the ground were taken into account. The nutritional state was evaluated at 60, 120, 180 and 240 days after the first application of the treatments, by collecting four ripe leaves of the branch before inflorescence, in all quadrants at the medium position in the plant. To evaluate fertility of the soil, some samples were collected at the 120th, 180th and 240th day at the depths 0 - 20 and 20 - 40 cm. According to the results, the following conclusions were drawn: the doses 148.06 and 30 g plant-1 of K2O and P2O5, respectively, resulted the highest fruit yield (84.29 kg plant-1); the leaf collections to quantify the contents of P and K in the hog-plum plant should be performed at 240 days; the highest contents of P and K in the leaves were reached, when the highest doses of those elements were applied on the soil; the hog-plum plant positively responds to the organic fertilization; both phosphorus and potassium fertilizations increased the levels of potassium in the soil at depths from 0 to 20 and from 20 to 40 cm. / A exploração extrativista, a irregularidade na produção e a falta de uma proposta de manejo da cajazeira (Spondias mombin L.), quanto aos aspectos fitotécnicos, colocam esta cultura numa condição marginal na cadeia produtiva. Assim, com o objetivo de identificar e quantificar as exigências nutricionais da cajazeira (Spondias mombin L.) para os nutrientes fósforo e potássio, foi realizado na estação experimental da Empresa de Pesquisa Agropecuária da Paraíba, EMEPA-PB, localizada no município de Mangabeira, um experimento em blocos casualizados com os tratamentos dispostos em esquema fatorial 4 x 4 + 2 , sendo os dois primeiros fatores constituídos pelas doses de P e K (30, 60, 120 e 240 g planta-1), acrescidos de duas testemunhas (sem adubação e adubação orgânica). Para a estimativa da variável produção, foram considerados apenas os frutos caídos ao chão. O estado nutricional foi avaliado aos 60, 120, 180 e 240 dias após a primeira aplicação dos tratamentos, com a coleta de 4 folhas maduras do ramo anterior à inflorescência, em todos os quadrantes, na posição mediana da planta. Para avaliar a fertilidade do solo, foram coletadas amostras aos 120, 180 e 240 dias nas profundidades de 0 - 20 e de 20 - 40 cm. Face aos resultados obtidos, concluiu-se que: as doses de 148,06 e 30 g planta-1 de K2O e P2O5, respectivamente, resultaram nas maiores produções de frutos (84,29 kg planta- 1); a coleta de folhas para quantificar os teores de P e K na cajazeira deve ser aos 240 dias; os maiores teores foliares para o P e K foram obtidos quando aplicadas as maiores doses dos respectivos elementos no solo; a cajazeira responde positivamente à adubação orgânica; e, a adubação fosfática e potássica elevaram os níveis de fósforo e potássio no solo nas profundidades de 0 a 20 e de 20 a 40 cm.

In this thesis, a novel Pedestrian Protection System (PPS), composed of the Pedestrian Detection System (PDS) and the Pedestrian Tracking System (PTS), was proposed. The PPS is a supplementary application for the Advanced Driver Assistance System, which is used to avoid collisions between vehicles and pedestrians. The Pedestrian Detection System (PDS) is used to detect pedestrians from near to far ranges with the feature-classi er-based detection method (HOG + SVM). To achieve pedestrian detection from near to far ranges, a novel structure was proposed. The structure of our PDS consists of two cameras (called CS and CL separately). The CS is equipped with a short focal length lens to detect pedestrians in near-to-mid range; and, the CL is equipped with a long focal length lens to detect pedestrians in mid-to-far range. To accelerate the processing speed of pedestrian detection, the parallel computing capacity of GPU was utilized in the PDS. The synchronization algorithm is also introduced to synchronize the detection results of CS and CL. Based on the novel pedestrian detection structure, the detection process can reach a distance which is more than 130 meters away without decreasing detection accuracy. The detection range can be extended more than 100 meters without decreasing the processing speed of pedestrian detection. Afterwards, an algorithm to eliminate duplicate detection results is proposed to improve the detection accuracy. The Pedestrian Tracking System (PTS) is applied following the Pedestrian Detection System. The PTS is used to track the movement trajectory of pedestrians and to predict the future motion and movement direction. A C + + class (called pedestrianTracking class, which is short for PTC) was generated to operate the tracking process for every detected pedestrian. The Kalman lter is the main algorithm inside the PTC. During the operation of PPS, the nal detection results of each frame from PDS will be transmitted to the PTS to enable the tracking process. The new detection results will be used to update the existing tracking results in the PTS. Moreover, if there is a newly detected pedestrian, a new process will be generated to track the pedestrian in the PTS. Based on the tracking results in PTS, the movement trajectory of pedestrians can be obtained and their future motion and movement direction can be predicted. Two kinds of alerts are generated based on the predictions: warning alert and dangerous alert. These two alerts represent di erent situations; and, they will alert drivers to the upcoming situations. Based on the predictions and alerts, the collisions can be prevented e ectively. The safety of pedestrians can be guaranteed.

In this thesis deep convolutional neural networks models and feature descriptor models are compared. Feature descriptors are paired with suitable chosen classifier. These models are a part of machine learning therefore machine learning types are described in this thesis. Further these chosen models are described, and their basics and problems are explained. Hardware and software used for tests is listed and then test results and results summary is listed. Then comparison based on the validation accuracy and training time of these said models is done.

Master's thesis studies the possibilities of generating 3D trees using variety of methods including context-free grammars and L-systems. Master's thesis also includes chapter on evolutionary and genetic algorithms, which briefly summarize their function. In this project genetic algorithm which takes 2D image of tree and the beginning of its trunk is proposed. Based on this information it will generate 3D tree which is visually close to the original image. In addition to methods of generating trees, reader will get information about processing input image and designing test application.

This study aims to modify Vision Transformer (ViT) to achieve higher accuracy. ViT is a model used in computer vision to, among other things, classify images. By applying ViT to the MNIST data set, an accuracy of approximately 98% is achieved. ViT is modified by implementing a method called Histogram of Oriented Gradients (HOG) in two different ways. The results show that the first approach with HOG gives an accuracy of 98,74% (setup 1) and the second approach gives an accuracy of 96,87% (patch size 4x4 pixels). The study shows that when HOG is applied on the entire image, a better accuracy is obtained. However, no systematic optimization has taken place, which makes it difficult to draw conclusions with certainty.

本研究主要在於設計一種新式生物反應器，並應用於生產豬瘟病毒疫苗。首先根據所培養細胞的生長特性與原有生物反應器之缺點，改良成新式的生物反應器，並評估此新式生物反應器適用性、效能，以及所培養豬腎細胞之生長代謝情形與豬瘟病毒力價。整個實驗過程大致分為兩個部分，第一個部分探討細胞固定化培養之最適化培養條件與生長代謝情形，第二個部分探討豬瘟病毒培養之最適化培養條件與生長代謝情形。實驗結果發現豬腎細胞(PK-15)以批次方式培養於新式生物反應器，搭配著FIBRA-CEL®載體，成功的進行擴大培養，豬腎細胞最高的生長量達到2.29×109cells/300mL的細胞量。因此，改良之新式生物反應器可提供細胞優越的生長環境，具有擴大規模培養之潛力，可藉由此簡單設備、操作容易、成本低且低能源消耗之新式生物反應器達成細胞製品之生產基座。 / In this study, the production of PK-15 cell using immobilized animal cell culture in a novel bioreactor was investigated.We evaluated the serviceability and efficiency of a design-improved novel bioreactor for the growth and metabolic states of cultured PK-15 cells and the production of HC virus. The entire experiment includes two major stages: (1) investigation of the optimal conditions and metabolic states for the growth of immobilized cells, (2) investigation of the optimal conditions for the production of HC virus. Our results showed that immobilized PK-15 cells on the fibra-cell carries in the design-improved novel bioreactor exhibited their best growth of 2.29×109 cells/300mL.The immobilized conditions for cell culture, can provide a shearing stress of growth state, easy separation of cells from the culture mediu, and a operation of continuously feeding medium, leading to possibility growth of the high density cell and a long period of production;as a result, the efficiency of producing process is promoted. Here,our design-improved novel bioreactor is expected to provide an optimal growth environment of both the cells and viruses for the production of high-yielded, stable, and consistent cellular biological preparations. Furthermore, it will also provide the basis for the production of cell products with advantages of simple-equipped, easy-to-operate, low cost, and low energy consumption. / 致謝 i 中文摘要 ------------------------------------------------------------------------ ii 英文摘要 ------------------------------------------------------------------------ iii 目錄 ------------------------------------------------------------------------ iv 表目錄 ------------------------------------------------------------------------ vi 圖目錄 ------------------------------------------------------------------------ vii 第一章 緒論------------------------------------------------------------------ 1 第二章 文獻回顧------------------------------------------------------------ 3 2-1 豬腎傳代細胞(PK-15 cell) -------------------------------------- 3 2-2 豬瘟病毒------------------------------------------------------------ 4 2-2.1 豬瘟之歷史背景--------------------------------------------------- 4 2-2.2 豬瘟病毒之特性--------------------------------------------------- 7 2-2.3 豬瘟發生原因之探討--------------------------------------------- 11 2-3 生物反應器--------------------------------------------------------- 12 第三章 實驗材料與方法--------------------------------------------------- 21 3-1 細胞------------------------------------------------------------------ 21 3-2 細胞繼代培養------------------------------------------------------ 21 3-3 細胞冷凍保存------------------------------------------------------ 22 3-4 解凍細胞培養------------------------------------------------------ 22 3-5 病毒感染------------------------------------------------------------ 23 3-6 收集病毒------------------------------------------------------------ 24 3-7 豬瘟病毒力價測試------------------------------------------------ 24 3-8 細胞滾瓶培養------------------------------------------------------ 26 3-9 生物反應器操作--------------------------------------------------- 27 3-10 載體上細胞數的測定--------------------------------------------- 33 3-11 葡萄糖的測定------------------------------------------------------ 33 3-12 培養過程中pH值測定------------------------------------------- 34 第四章 結果與討論--------------------------------------------------------- 35 4-1 測試細胞貼附的材料--------------------------------------------- 35 4-2 細胞固定時間的比較--------------------------------------------- 36 4-3 測試不同比例的載體量培養豬腎細胞------------------------ 37 4-4 測試不同接種量--------------------------------------------------- 40 4-5 測試培養基流速對豬腎細胞生長的影響--------------------- 44 4-6 測試培養基停留於培養槽時間對豬腎細胞生長的影響--- 45 4-7 測試豬腎細胞暴露空氣時間對於生長的影響--------------- 47 4-8 測試Bellocell培養豬腎細胞(PK-15)可行性----------------- 49 4-9 測試利用新式生物反應器培樣豬瘟病毒--------------------- 50 第五章 結論與建議--------------------------------------------------------- 53 參考文獻 ------------------------------------------------------------------------ 55 表目錄 表1. 兔化豬瘟疫苗與組織培養豬瘟疫苗的比較------------------ 6 表2. 急性、慢性與遲發型豬瘟比較---------------------------------- 10 表3. .Growth of Various cell Lines in bellocell-500----------------- 18 表4. Comparison of SF-9 cell Growth and BEV production in Various Laboratory bioreators------------------------------------ 19 表5. Comparison of HEK293 Cell growth and Receptor X production in Cell Factory®/20 roller bottles and BelloCell-500Bioreactor------------------------------------------ 20 表6. Reed-Muench Methods法計算方法----------------------------- 26 表7. 比較不同材料培養PK-15 cell所用的載體量---------------- 50 表8. 細胞固定時間的比較所接細胞量與載體量------------------ 51 圖目錄 圖1. Liau提出以潮汐生物反應器圖--------------------------------- 17 圖2. Operation principle of Bellocell system------------------------- 18 圖3. 新式生物反應器(novel reactor)-潮汐式生物反應器(tidal typereactor)之運作流程圖--------------------------------------- 30 圖4. 比較不同材料培養PK-15 cell ---------------------------------- 63 圖5. 比較不同時間細胞的貼附量------------------------------------ 64 圖6. 測試的不同比例載體量培養豬腎細胞生長曲線------------ 65 圖7. 測試的不同比例載體量培養豬腎細胞培養過程glucose消耗趨勢------------------------------------------------------------ 66 圖8. 測試的不同比例載體量培養豬腎細胞培養過程pH變化------------------------------------------------------------------------ 67 圖9. 測試不同接細胞量培養在10g carrier生長曲線------------- 68 圖10. 測試不同接細胞量培養在10g 載體glucose消耗趨勢----- 69 圖11. 測試不同接細胞量培養在10g carrier pH趨勢--------------- 70 圖12. 測試流速對豬腎細胞生長的影響------------------------------ 71 圖13. 測試流速對細胞影響的葡萄糖消耗--------------------------- 72 圖14. 測試流速對豬腎細胞生長影響pH值-------------------------- 73 圖15. 測試培養基停留時間對豬腎細胞生長影響------------------ 74 圖16. 測試培養基holding時間對豬腎細胞生長的影響之葡萄糖趨勢--------------------------------------------------------------- 75 圖17. 測試培養基holding時間對細胞的影響之pH值趨勢------- 76 圖18. 測試豬腎細胞暴露空氣時間對生長的影響------------------ 77 圖19. 測試豬腎細胞暴露空氣對生長的影響葡萄糖消耗趨勢--- 78 圖20. 測試豬腎細胞暴露空氣對細胞生長的影養pH值趨勢---- 79 圖21. Bellocell反應器培養豬腎細胞---------------------------------- 80 圖22. Bellocell培養豬腎細胞葡萄糖消耗趨勢---------------------- 81 圖23. Bellocell培養豬腎細胞pH值趨勢----------------------------- 82 圖24. 測試利用新式生物反應器培養豬瘟病------------------------ 83