Submitted by Maurício Gianfrancesco Filippi null (mauriciofilippi@terra.com.br) on 2017-02-13T18:05:19Z No. of bitstreams: 1 Dissertação Maurício G Filippi oficial.pdf: 3286618 bytes, checksum: 2e08ac3322153696d51497bfacf1f0f8 (MD5) / Rejected by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo a orientação abaixo: O arquivo submetido está sem a ficha catalográfica. A versão submetida por você é considerada a versão final da dissertação/tese, portanto não poderá ocorrer qualquer alteração em seu conteúdo após a aprovação. Corrija esta informação e realize uma nova submissão com o arquivo correto. Agradecemos a compreensão. on 2017-02-15T17:11:59Z (GMT) / Submitted by Maurício Gianfrancesco Filippi null (mauriciofilippi@terra.com.br) on 2017-02-16T22:41:18Z No. of bitstreams: 1 Dissertação Maurício G Filippi.pdf: 3283472 bytes, checksum: fa27139485cf995a23ebdca2e069dd69 (MD5) / Approved for entry into archive by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br) on 2017-02-20T18:00:33Z (GMT) No. of bitstreams: 1 fillipi_mg_me_bot.pdf: 3283472 bytes, checksum: fa27139485cf995a23ebdca2e069dd69 (MD5) / Made available in DSpace on 2017-02-20T18:00:33Z (GMT). No. of bitstreams: 1 fillipi_mg_me_bot.pdf: 3283472 bytes, checksum: fa27139485cf995a23ebdca2e069dd69 (MD5) Previous issue date: 2016-12-12 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A monitorização eletrocardiográfica ambulatorial, ou método Holter, vem se mostrando como uma ferramenta eficaz na Medicina Veterinária para detectar lesões cardíacas precoces, não só por monitorar a atividade elétrica do coração, como também o controle do sistema nervoso autônomo desse órgão. Sabe-se também que as principais enfermidades infecciosas de cães, como a cinomose e a erliquiose monocítica canina (EMC) provocam lesões consideráveis no coração, comprovadas por exame histopatológico. Já está comprovada a ocorrência de miocardite na EMC, levando a frequente presença de alterações na geração e condução do impulso elétrico cardíaco. O presente estudo analisou a atividade elétrica do coração durante 24 horas, com enfoque na prevalência de arritmias, estudo da variabilidade da frequência cardíaca e na concentração de biomarcadores de cães com EMC crônica (grupo doente) em comparação à animais saudáveis (grupo controle). Quarenta e cinco por cento dos animais do grupo doente possuíram alta frequência de arritmias durante o estudo. A média da concentração de troponina cardíaca I e de creatinokinase MB (CK-MB) foi significativa (0,24 ng/mL ± 0,5; 229 ± 205 UI/mL) em comparação ao grupo controle (0,042 ± 0,07 ng/mL; 126 ± 46,12 UI/mL). O desvio padrão da média de todos os intervalos NN (SDNN) e a porcentagem de intervalos RR adjacentes com diferença de duração superior a 50 milissegundos (pnn50%) também foram extremamente singificativos (83 ± 65 e 14,56 ± 20) quando comparado aos animais saudáveis (268 ± 74,6 e 55,87 ± 12,8), respectivamente. Podemos concluir que a EMC crônica possui caráter arritmôgenico, onde há persistente lesão miocárdica e intensa estimulação do sistema nervoso autônomo simpático no coração. / Ambulatorial electrocardiographic monitoring, or Holter method, has been shown to be an effective tool in veterinary medicine to detect early heart lesions, not only to monitor the electrical activity of the heart, but also to control the autonomic nervous system of this organ. It is also known that the main infectious diseases of dogs, such as canine distemper and canine monocytic ehrlichiosis (CME) cause considerable lesions in the heart, proven by histopathological examination. It has already been proven the occurrence of myocarditis in the CME, leading to frequent presence of changes in the generation and conduction of the cardiac electrical impulse. The present study analyzed the electrical activity of the heart during 24 hours, focusing on the prevalence of arrhythmias, heart rate variability study and the biomarkers concentration of dogs with chronic CME (sick group) compared to healthy animals (control group). Forty-five percent of the animals in the diseased group had a high frequency of arrhythmias during the study. The mean concentration of cardiac troponin I and creatinokinase MB (CK-MB) was significant (0.24 ng / mL ± 0.5; 229 ± 205 IU / mL) compared to the control group (0.042 ± 0.07 ng / ML, 126 ± 46.12 IU / mL). The standard deviation of the mean of all NN (SDNN) intervals and the percentage of adjacent RR intervals with a duration difference greater than 50 milliseconds (pnn50%) were also extremely significant (83 ± 65 and 14.56 ± 20) when compared to Healthy animals (268 ± 74.6, 55.87 ± 12.8), respectively. It can be concluded that chronic CME has an arrhythmogenic character, where there is persistent myocardial injury and intense stimulation of the sympathetic autonomic nervous system of the heart. / FAPESP: 2014/11219-6

Hepatocyte growth factor (HGF) and its specific receptor MET play a role in many physiological functions including proliferation, migration and morphogenesis. Recently, research results in our laboratory showed that recombinant HGF variants (NK1, NK2, NK3 and NK4) became antagonists to HGF/MET pathway by suppressing proliferation, migration and invasion in human breast cancer cells (MDA-MB-435S, MDA). Similar results were achieved when HGF variants genes were introduced in MDA cells. To understand the molecular mechanism of breast cancer cells metastasis suppressed by HGF variants, MDA and five transfectants, including MDA-GFP, MDA-NK1, MDA-NK2, MDA-NK3 and MDA-NK4 cells were used for proteomic analysis using two-dimensional electrophoresis (2-DE). Differential analysis revealed that a total of 56 polypeptides were differentially expressed through five sets of comparison using wild-type MDA cells as a control. A total of 17 polypeptides were shown differential expression between MDA and MDA-GFP cells, with 11 down-regulated and 6 up-regulated. Eighteen polypeptides were differentially expressed between MDA and MDA-NK1 cells, with 15 down-regulated and 3 up-regulated. There were 22 differentially expressed polypeptides found between MDA and MDA-NK2 cells, in which 14 were down-regulated and 8 were up-regulated. Sixteen polypeptides were shown differentially expressed between MDA and MDA-NK3 cells, with 11 down-regulated and 5 up-regulated. A total of 18 polypeptides were shown differential expression between MDA and MDA-NK4 cells, with 15 down-regulated and 3 up-regulated. Proteomic analysis showed that a total of 43 polypeptides were differentially expressed through four sets of comparison (MDA-GFP and MDA-NK1, MDA-GFP and MDA-NK2, MDA-GFP and MDA-NK3, and MDA-GFP and MDA-NK4). To understand the differential expression among different HGF variants-transfected MDA cells, three sets of cross analysis were also carried out (MDA-NK1 and MDA-NK2, MDA-NK1 and MDA-NK3, and MDA-NK1 and MDA-NK4) and the results showed that a total of 37 differentially expressed polypeptides were found in the three sets of comparison. Similarly, when MDA-NK2 cells were used as a control to compare with MDA-NK3 and MDA-NK4 cells, 34 significantly differential expressed polypeptides were found. The last set of comparison between MDA-NK3 and MDA-NK4 cells, 19 polypeptides were found significantly differential expression. Therefore, our current results revealed that the differentially expressed polypeptides in MDA-MB-435S cells and HGF variants-transfected MDA cells could be related to the inhibition of proliferation and migration of human breast cancer cells by HGF variants.

The allocation of frequency spectrum by the FCC for Ultra Wideband (UWB) communications in the 3.1-10.6 GHz has paved the path for very high data rate Gb/s wireless communications. Frequency synthesis in these communication systems involves great challenges such as high frequency and wideband operation in addition to stringent requirements on frequency hopping time and coexistence with other wireless standards. This research proposes frequency generation schemes for such radio systems and their integrated implementations in silicon based technologies. Special emphasis is placed on efficient frequency planning and other system level considerations for building compact and practical systems for carrier frequency generation in an integrated UWB radio. This work proposes a frequency band plan for multiband OFDM based UWB radios in the 3.1-10.6 GHz range. Based on this frequency plan, two 11-band frequency synthesizers are designed, implemented and tested making them one of the first frequency synthesizers for UWB covering 78% of the licensed spectrum. The circuits are implemented in 0.25µm SiGe BiCMOS and the architectures are based on a single VCO at a fixed frequency followed by an array of dividers, multiplexers and single sideband (SSB) mixers to generate the 11 required bands in quadrature with fast hopping in much less than 9.5 ns. One of the synthesizers is integrated and tested as part of a 3-10 GHz packaged receiver. It draws 80 mA current from a 2.5 V supply and occupies an area of 2.25 mm2. Finally, an architecture for a UWB synthesizer is proposed that is based on a single multiband quadrature VCO, a programmable integer divider with 50% duty cycle and a single sideband mixer. A frequency band plan is proposed that greatly relaxes the tuning range requirement of the multiband VCO and leads to a very digitally intensive architecture for wideband frequency synthesis suitable for implementation in deep submicron CMOS processes. A design in 130nm CMOS occupies less than 1 mm2 while consuming 90 mW. This architecture provides an efficient solution in terms of area and power consumption with very low complexity.

In a pursuance of a small-scale field experiment was to monitor the impact of extrarooted nutrition as one of the factors which influence the yield and technological quality of sugar beet. In the course of the vegetation was monitored for the growth and development of diabetes and the dynamics of change of its quality. On the basis of the observed results were evaluated by the following parameters - yield bulev, the yield shaw, sugar content, soluble ash, alpha-amino-nitrogen and MB factor. The highest yield of root was recorded after the application of fertilizers Yara Vita Bortrac (127 t/ha). At the time of harvest was the highest yield shaw determined after the application of the product Carbonbor On (39,6 t/ha). The highest sugar content was determined in the two years after the fertilization of the product Carbonbor. In the average of the two years has moved the content of soluble ash in the range of 0,68 - 0,75 %. The lowest was at the variants with the application of Yara Vita Brassitrel and highest in the variant, after the application of fertilizers Carbonbor K. The content of alpha-amino-nitrogen in different years differed, in the year 2012 amounted to the average of the value of 36,7 mg.100 g-1, and in 2013 was 26.3 mg.100 g-1. The highest value of the MB factor was determined for the variants with the application of fertilizers 3 Carbonbor K. Also there has been achieved a favorable value of 27. Most mature sugar beet has been harvested after the application of fertilizers Carbonbor.

abstract: Breast cancer cell invasion is a highly orchestrated process driven by a myriad of complex microenvironmental stimuli. These complexities make it difficult to isolate and assess the effects of specific parameters including matrix stiffness and tumor architecture on disease progression. In this regard, morphologically accurate tumor models are becoming instrumental to perform fundamental studies on cancer cell invasion within well-controlled conditions. In this study, the use of photocrosslinkable hydrogels and a novel, two-step photolithography technique was explored to microengineer a 3D breast tumor model. The microfabrication process presented herein enabled precise localization of the cells and creation of high stiffness constructs adjacent to a low stiffness matrix. To validate the model, breast cancer cell lines (MDA-MB-231, MCF7) and normal mammary epithelial cells (MCF10A) were embedded separately within the tumor model and cellular proliferation, migration and cytoskeletal organization were assessed. Proliferation of metastatic MDA-MB-231 cells was significantly higher than tumorigenic MCF7 and normal mammary MCF10A cells. MDA-MB-231 exhibited highly migratory behavior and invaded the surrounding matrix, whereas MCF7 or MCF10A cells formed clusters that were confined within the micropatterned circular features. F-actin staining revealed unique 3D protrusions in MDA-MB-231 cells as they migrated throughout the surrounding matrix. Alternatively, there were abundance of 3D clusters formed by MCF7 and MCF10A cells. The results revealed that gelatin methacrylate (GelMA) hydrogel, integrated with the two-step photolithography technique, has great promise in creating 3D tumor models with well-defined features and tunable stiffness for detailed studies on cancer cell invasion and drug responsiveness. / Dissertation/Thesis / Supplementary Movie 3 / Supplementary Movie 1 / Supplementary Movie 2 / Supplementary Movie 5 / Supplementary Movie 4 / Masters Thesis Bioengineering 2015

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