As células mesenquimais estromais multipotentes (CMM) são na atualidade uma fonte atrativa para aplicações na engenharia de tecidos e na terapia celular. Devido à baixa disponibilidade nos tecidos (0,01%-0,0005%) e às elevadas doses necessárias para uma infusão (aproximadamente 106 células/Kg paciente) tornou-se necessário o desenvolvimento de tecnologias de expansão in vitro, eficientes e de custo reduzido, que permitam a obtenção de CMM com manutenção das características funcionais (diferenciação e inibição da proliferação de linfócitos), imunofenotípicas e citogenéticas. As CMM são células aderentes, ou seja, necessitam de um substrato sólido para se aderir e proliferar. O procedimento convencional de expansão em garrafas estáticas, geralmente envolve um processo laborioso em que não há correto controle e monitoramento dos parâmetros de cultivo e possui uma maior susceptibilidade à contaminação devido à excessiva manipulação para atingir o número ideal de células. Além disso, este tipo de cultivo não permite uma produção em larga escala. Em função disso, o presente trabalho foi proposto com o objetivo de desenvolver um bioprocesso escalonável, economicamente viável e eficiente para expansão de CMM derivadas da medula óssea em microcarregadores. Para isso, as células foram cultivadas em microcarregador Cyotdex 3, em frasco spinner com o meio -MEM suplementado com 15% de SFB. Foram avaliadas neste trabalho, a adesão celular aos microcarregadores, crescimento, metabolismo, recuperação celular final e avaliação das propriedades funcionais e imunofenotípicas pré e pós cultivo, comparando ao cultivo já estabelecido em garrafas estáticas. De maneira geral, os resultados obtidos mostraram que foi possível expandir CMM utilizando a tecnologia de microcarregadores. A análise do metabolismo celular mostrou que não houve exaustão de nutrientes importantes como glicose e glutamina durante o cultivo, tampouco formação dos subprodutos lactato e amônia em concentrações inibitórias. As células recuperadas após a expansão mantiveram as características imunofenotípicas e funcionais. A produção média (n=10) foi de aproximadamente 4,9x105 cel/mL. Como o sistema utilizado permite o escalonamento, se utilizássemos um biorreator de 1L, seria possível a produção de aproximadamente 5x108 células que seriam suficientes para tratar mais de 3 pacientes de até 70Kg na dose de 2x106 células/Kg. Para expansão da mesma quantidade de células na forma tradicional seriam necessárias 135 garrafas de 175 cm2 com um custo total de expansão duas vezes superior à estimativa do custo de expansão utilizando microcarregadores. / Multipotent mesenchymal stromal cells are currently an attractive source for applications in tissue engineering and cell therapy. Due to the low availability in tissues (0,01%-0,0005%) and the high doses necessary for an infusion (about 106 cells/Kg patient), it has become necessary the development of effective and low cost technologies for in vitro expansion that enable to obtain MSC with maintenance of functional (differentiation and inhibition of lymphocytes proliferation), immunophenotypic and cytogenetics characteristics. MSC are adherent cells, i.e., they need a solid substrate to adhere and proliferate. The conventional procedure for expansion in static flasks normally involves a laborious process in which there is no suitable control and monitoring of the cultivation parameters besides presenting a higher susceptibility to contamination due to excessive manipulation to reach the ideal amount of cells. Moreover, this kind of cultivation does not allow a large scale production. For this reason, this work was proposed with the objective to develop a low cost, effective and scalable bioprocess for expansion of bone marrow-derived MSC in microcarriers. Cells grew on microcarriers Cyotdex 3, in spinner flasks with the -MEM medium supplemented with 15% FBS. We evaluated the cell adhesion to microcarriers, growth, metabolism, final cell recovery, and the functional and immunophenotypic properties before and after cultivation, comparing them with the cultivation already established in static flasks. In general, the results obtained showed that it was possible to expand MSC using microcarriers technology. The analysis of the cell metabolism showed that there was no depletion of important nutrients such as glucose and glutamine during cultivation, neither formation of lactate and ammonia subproducts in inhibitory concentrations. The cells recovered after the expansion kept the immunophenotypic and functional characteristics. The mean production (n=10) was about 4,9x105 cel/mL. As the system used allows the scale-up, if we had used a bioreactor of 1L it would had been possible to produce approximately 5x108 cells that would be enough to treat more than three patients of up to 70kg with a dose of 2x106 cells/kg. For the expansion of the same amount of cells in the traditional way, it would be necessary 135 T-flasks of 175 cm2 with total cost twice higher than the estimate cost of expansion using microcarriers.

As células mesenquimais estromais multipotentes (CMM) são na atualidade uma fonte atrativa para aplicações na engenharia de tecidos e na terapia celular. Devido à baixa disponibilidade nos tecidos (0,01%-0,0005%) e às elevadas doses necessárias para uma infusão (aproximadamente 106 células/Kg paciente) tornou-se necessário o desenvolvimento de tecnologias de expansão in vitro, eficientes e de custo reduzido, que permitam a obtenção de CMM com manutenção das características funcionais (diferenciação e inibição da proliferação de linfócitos), imunofenotípicas e citogenéticas. As CMM são células aderentes, ou seja, necessitam de um substrato sólido para se aderir e proliferar. O procedimento convencional de expansão em garrafas estáticas, geralmente envolve um processo laborioso em que não há correto controle e monitoramento dos parâmetros de cultivo e possui uma maior susceptibilidade à contaminação devido à excessiva manipulação para atingir o número ideal de células. Além disso, este tipo de cultivo não permite uma produção em larga escala. Em função disso, o presente trabalho foi proposto com o objetivo de desenvolver um bioprocesso escalonável, economicamente viável e eficiente para expansão de CMM derivadas da medula óssea em microcarregadores. Para isso, as células foram cultivadas em microcarregador Cyotdex 3, em frasco spinner com o meio -MEM suplementado com 15% de SFB. Foram avaliadas neste trabalho, a adesão celular aos microcarregadores, crescimento, metabolismo, recuperação celular final e avaliação das propriedades funcionais e imunofenotípicas pré e pós cultivo, comparando ao cultivo já estabelecido em garrafas estáticas. De maneira geral, os resultados obtidos mostraram que foi possível expandir CMM utilizando a tecnologia de microcarregadores. A análise do metabolismo celular mostrou que não houve exaustão de nutrientes importantes como glicose e glutamina durante o cultivo, tampouco formação dos subprodutos lactato e amônia em concentrações inibitórias. As células recuperadas após a expansão mantiveram as características imunofenotípicas e funcionais. A produção média (n=10) foi de aproximadamente 4,9x105 cel/mL. Como o sistema utilizado permite o escalonamento, se utilizássemos um biorreator de 1L, seria possível a produção de aproximadamente 5x108 células que seriam suficientes para tratar mais de 3 pacientes de até 70Kg na dose de 2x106 células/Kg. Para expansão da mesma quantidade de células na forma tradicional seriam necessárias 135 garrafas de 175 cm2 com um custo total de expansão duas vezes superior à estimativa do custo de expansão utilizando microcarregadores. / Multipotent mesenchymal stromal cells are currently an attractive source for applications in tissue engineering and cell therapy. Due to the low availability in tissues (0,01%-0,0005%) and the high doses necessary for an infusion (about 106 cells/Kg patient), it has become necessary the development of effective and low cost technologies for in vitro expansion that enable to obtain MSC with maintenance of functional (differentiation and inhibition of lymphocytes proliferation), immunophenotypic and cytogenetics characteristics. MSC are adherent cells, i.e., they need a solid substrate to adhere and proliferate. The conventional procedure for expansion in static flasks normally involves a laborious process in which there is no suitable control and monitoring of the cultivation parameters besides presenting a higher susceptibility to contamination due to excessive manipulation to reach the ideal amount of cells. Moreover, this kind of cultivation does not allow a large scale production. For this reason, this work was proposed with the objective to develop a low cost, effective and scalable bioprocess for expansion of bone marrow-derived MSC in microcarriers. Cells grew on microcarriers Cyotdex 3, in spinner flasks with the -MEM medium supplemented with 15% FBS. We evaluated the cell adhesion to microcarriers, growth, metabolism, final cell recovery, and the functional and immunophenotypic properties before and after cultivation, comparing them with the cultivation already established in static flasks. In general, the results obtained showed that it was possible to expand MSC using microcarriers technology. The analysis of the cell metabolism showed that there was no depletion of important nutrients such as glucose and glutamine during cultivation, neither formation of lactate and ammonia subproducts in inhibitory concentrations. The cells recovered after the expansion kept the immunophenotypic and functional characteristics. The mean production (n=10) was about 4,9x105 cel/mL. As the system used allows the scale-up, if we had used a bioreactor of 1L it would had been possible to produce approximately 5x108 cells that would be enough to treat more than three patients of up to 70kg with a dose of 2x106 cells/kg. For the expansion of the same amount of cells in the traditional way, it would be necessary 135 T-flasks of 175 cm2 with total cost twice higher than the estimate cost of expansion using microcarriers.

Embryonic stem (ES) cells provide a potentially unlimited cell source for cellular therapies; however, reliable methods must be developed to provide clinically-relevant numbers of homogeneous therapeutic cell populations. Dynamic cultures may encourage ES cell differentiation and amenable to large-scale cell production. Our goal was to optimize dynamic culture parameters (bioreactor type, speed, cell seeding density, conditioned medium, and hypoxia) to maximize the generation of hematopoietic stem and progenitor cells (HSPCs) from ES cells and also to investigate the ability of dynamic culture-derived HSPCs to generate terminally differentiated hematopoietic cells. Our results indicate that varying cell seeding density and speed in two different bioreactors significantly affects embryoid body formation and ES cell differentiation efficiency into progenitor cells. In general, increased cell seeding density generated higher percentages of HSPCs in both bioreactors. In addition, rotary (Synthecon) bioreactors produced more sca-1⁺ progenitors, and spinner flasks generated more c-kit⁺ progenitors, demonstrating their unique differentiation profiles. cDNA microarray analysis of genes involved in pluripotency, germ layer formation, and hematopoietic differentiation showed that unique gene expression profiles were observed in the two bioreactors with the expression of specific hematopoietic genes more up regulated in the Synthecon cultures compared to spinner flasks. Combining bioreactor cultures with directed differentiation strategies via conditioned medium and hypoxic culture may further encourage hematopoietic differentiation. Dynamically cultured ES cell-derived hematopoietic stem and progenitor cells were further differentiated into a phenotype typical of dendritic cells which had the ability to process antigen. Additionally, microarray analysis of isolated ES cell-derived HSPCs demonstrated differences in the gene expression from native HSCs isolated from the fetal liver or bone marrow of mice. Insight gained from this work should be continued by comparing the differentiation efficiency of HSPCs derived in dynamic and traditional static culture methods into functional, terminally differentiated hematopoietic cells to generate clinically-relevant numbers of transplantable, therapeutic cells. / text

Thesis (M.S.)--North Carolina State University, 2003. / Title from PDF t.p. (viewed on Aug. 21, 2004). Includes vita. Includes bibliographical references (p. 88-94).

<p>Sound is a key sensory modality for Hawaiian spinner dolphins. Like many other marine animals, these dolphins rely on sound and their acoustic environment for many aspects of their daily lives, making it is essential to understand soundscape in areas that are critical to their survival. Hawaiian spinner dolphins rest during the day in shallow coastal areas and forage offshore at night. In my dissertation I focus on the soundscape of the bays where Hawaiian spinner dolphins rest taking a soundscape ecology approach. I primarily relied on passive acoustic monitoring using four DSG-Ocean acoustic loggers in four Hawaiian spinner dolphin resting bays on the Kona Coast of Hawai‛i Island. 30-second recordings were made every four minutes in each of the bays for 20 to 27 months between January 8, 2011 and March 30, 2013. I also utilized concomitant vessel-based visual surveys in the four bays to provide context for these recordings. In my first chapter I used the contributions of the dolphins to the soundscape to monitor presence in the bays and found the degree of presence varied greatly from less than 40% to nearly 90% of days monitored with dolphins present. Having established these bays as important to the animals, in my second chapter I explored the many components of their resting bay soundscape and evaluated the influence of natural and human events on the soundscape. I characterized the overall soundscape in each of the four bays, used the tsunami event of March 2011 to approximate a natural soundscape and identified all loud daytime outliers. Overall, sound levels were consistently louder at night and quieter during the daytime due to the sounds from snapping shrimp. In fact, peak Hawaiian spinner dolphin resting time co-occurs with the quietest part of the day. However, I also found that humans drastically alter this daytime soundscape with sound from offshore aquaculture, vessel sound and military mid-frequency active sonar. During one recorded mid-frequency active sonar event in August 2011, sound pressure levels in the 3.15 kHz 1/3rd-octave band were as high as 45.8 dB above median ambient noise levels. Human activity both inside (vessels) and outside (sonar and aquaculture) the bays significantly altered the resting bay soundscape. Inside the bays there are high levels of human activity including vessel-based tourism directly targeting the dolphins. The interactions between humans and dolphins in their resting bays are of concern; therefore, my third chapter aimed to assess the acoustic response of the dolphins to human activity. Using days where acoustic recordings overlapped with visual surveys I found the greatest response in a bay with dolphin-centric activities, not in the bay with the most vessel activity, indicating that it is not the magnitude that elicits a response but the focus of the activity. In my fourth chapter I summarize the key results from my first three chapters to illustrate the power of multiple site design to prioritize action to protect Hawaiian spinner dolphins in their resting bays, a chapter I hope will be useful for managers should they take further action to protect the dolphins.</p> / Dissertation

Sidladdningar på webbplatser har visat sig ha stor påverkan på hur användare beter sig på webbplatser. Redan efter 1 sekunds fördröjning börjar användarens beteende ändras. Laddningsindikatorer kan användas för att minska den upplevda hastigheten av sidladdningar. Det finns olika varianter av laddningsindikatorer som fungerar på olika sätt. En traditionell laddningsindikator är spinner och en nyare, mindre undersökt, heter skeleton screen.  Syftet med denna kandidatuppsats har varit att undersöka effektiviteten hos skeleton screen att påverka den upplevda hastigheten av sidladdningar. Dessutom var syftet att utforma och demonstrera ett test för att förse webbutvecklare med vägledning om hur liknande test kan genomföras i syfte att avgöra vilken laddningsindikator de bör välja att använda vid utveckling. En prototypisk webbsida skapades i två olika versioner, en med 2 sekunders fördröjning och en med 4 sekunders fördröjning vid sidladdning. För att ladda sidorna användes spinner eller skeleton screen. För att samla in användares upplevda hastighet av webbplatsen utfördes användartester där deltagarna för varje sida fick uppskatta hur lång tid nedladdningen tog. Totalt deltog 14 testpersoner i åldrarna 13–47 år. Resultatet visade på att skeleton screen påverkade testpersonernas upplevda hastighet som kortast i både versionerna. Medelvärdet av alla resultatet från testpersonerna visade på att vid 2 sekunders fördröjning upplevdes sidladdningar som 0.89 sekunder med skeleton screen och 1,95 sekunder med spinner. Vid 4 sekunders fördröjning upplevdes skeleton screen som 1,38 sekunder och spinner 3,81 sekunder. Trots att skeleton screen var den laddningsindikator som minskade upplevd hastighet mest, fanns det delade åsikter om vilken laddningsindikator testpersonerna föredrog baserat på design och tidigare erfarenheter. Skeleton screen verkar inte vara meningsfulla på sidor med enstaka produkter, utan effektiviteten av skeleton screen visades främst vid laddning av flera produkter.

The popularity of variable-rate technology (VRT) has grown. However, the limitations and errors ofthis technology are generally unknown. Therefore, a spatial data model was developed to generate "asapplied"surfaces to advance precision agricultural (PA) practices. A test methodology based on ASAEStandard S341.2 was developed to perform uniform-rate (UR) and variable-rate (VR) tests to characterizedistribution patterns testing four VRT granular applicators (two spinner spreaders and two pneumaticapplicators). Single-pass UR patterns exhibited consistent shapes for three of the applicators with patternsshifts observed for the fourth applicator. Simulated overlap analysis showed that three of the applicatorsperformed satisfactorily with most CVs less than 20% while one applicator performed poorly (CVs andgt;25%). The spinner spreaders over-applied at the margins but the pneumatic applicators under-appliedsuggesting a required adjustment to the effective swath spacing. Therefore, it is recommended that CVsaccompany overlap pattern plots to ensure proper calibration of VRT application.Quantification of the rate response characteristics for the various applicators illustrated varying delayand transition times. Only one applicator demonstrated consistent delay and transition times. A sigmoidalfunction was used to model the rate response for applicators. One applicator exhibited a linear responseduring a decreasing rate change. Rate changes were quicker for the two newer VR control systemssignifying advancement in hydraulic control valve technology. This research illustrates the need forstandard testing protocols for VRT systems to help guide VRT software developers, equipmentmanufacturers, and users.The spatial data model uses GIS functionality to merge applicator descriptive patterns with a spatialfield application file (FAF) to generate an 'as-applied' surface representing the actual distribution ofgranular fertilizer. Field data was collected and used to validate the "as-applied" spatial model.Comparisons between the actual and predicted application rates for several fields were madedemonstrating good correlations for one applicator (several R2 andgt; 0.70), moderate success for anotherapplicator (0.60 andlt; R2 andlt; 0.66), and poor relationships for the third applicator (R2 andlt; 0.49). A comparison ofthe actual application rates to the prescription maps generated R2 values between 0.16 and 0.81demonstrating inconsistent VRT applicator performance. Thus, "as-applied" surfaces provide a means toproperly evaluate VRT while enhancing researchers' ability to compare VR management approaches.

Made available in DSpace on 2016-08-17T18:39:40Z (GMT). No. of bitstreams: 1 3854.pdf: 3648790 bytes, checksum: 7cf8d19b11b10f9d50e9f3a326c8d0db (MD5) Previous issue date: 2010-10-07 / Universidade Federal de Sao Carlos / The Mesenchymal Stem Cells (MSCs) from adult tissues are at present an attractive source for applications in tissue engineering and cellular therapies. It presents a potential to differentiate into different cell lines as adipocytes, chondrocytes, osteocytes and myocytes. Due to low availability in the tissues and the fact that the demand for therapeutic applications is much larger than the supply capacity has become necessary to expand them in vitro strongly dependent on conservation of their differentiation potential. The MSCs are anchorage dependent, they have the characteristic of adhering to a solid substrate for subsequent proliferation. The conventional procedure of expansion in T-flasks with growth in monolayers, usually involves a difficult monitoring and control process, with high contamination indexi, low cell yield and high cost. In this work, we evaluated a methodology of expansion of hMSC-TERT line in spinner flask with microcarriers (small particles with properties favorable for cell adhesion and proliferation). In the experiments, two type of microcarriers were used, Cytodex 1 non-porous and with positive electric charge and Cultispher S, macroporous and electrically neutral, both in stirred spinner flask of 100 mL in culture medium &#945;-MEM with 15% fetal calf serum, maintained in a CO2 incubator at 37°C and pH between 7.2 and 7.4. To increase productivity, some strategies were adopted during the research as a nutritional balance of the culture medium, addition of microcarriers and dilution and/or replacement of medium during cultivation. Aided by analysis of metabolites by high performance liquid chromatography and optical microscopy, the results showed that it was possible to grow hMSC-TERT with two microcarriers while maintaining good conditions for growth in three dimensions. However, the cell yield was limited mainly by the formation of clusters of microcarriers/extracellular matrix and is possible to obtain a cellular expansion factor of 4.98 with Cytodex 1 and 9,70 with Cultispher S. Because of the easy recovery of the cell showed by later microcarrier, it was performed an analysis of the maintenance of antigenic phenotype by flow cytometry and induction of differentiation into adipocytes and osteocytes. The results confirmed the conversation of phenotypic characteristics of hMSC-TERT with the cultivation technique evaluated in this work. / As Células-Tronco Mesenquimais (CTMs) de tecido adulto são na atualidade uma fonte atrativa para aplicações na engenharia de tecidos e em terapias celulares. Apresentam um potencial de diferenciação em diversas linhagens celulares como adipócitos, condrócitos, osteócitos e miócitos. Devido à baixa disponibilidade nos tecidos e ao fato da demanda para aplicações terapêuticas ser muito maior do que a capacidade de fornecimento tornou-se necessária sua expansão in vitro fortemente condicionada à conservação do seu potencial de diferenciação. As CTMs são dependentes de ancoramento, ou seja, possuem a característica de aderir a um substrato sólido para posterior proliferação. O procedimento convencional de expansão em frascos T, com crescimento em monocamadas, geralmente envolve um processo de difícil monitoramento e controle, com maior índice de contaminação, de baixo rendimento celular e de alto custo. No presente trabalho, avaliou-se uma metodologia de expansão da linhagem hMSC-TERT em frasco spinner com microcarregadores (pequenas partículas com propriedades favoráveis para adesão e proliferação celular). Nos cultivos foram utilizados dois tipos de microcarregadores, Cytodex 1, não poroso e com carga elétrica positiva, e Cultispher S, macroporoso e eletricamente neutro, ambos em frasco spinner agitado de 100 mL, com o meio de cultura &#945;-MEM com 15% de soro fetal bovino, mantidos em incubadora de CO2 a 37ºC e pH entre 7,2 e 7,4. Para aumentar a produtividade celular, algumas estratégias foram adotadas durante a pesquisa como balanceamento nutricional do meio de cultura, adição de microcarregadores e a diluição e/ou a troca do meio durante o cultivo. Auxiliado pela análise de metabólitos por cromatografia líquida de alta eficiência e por microscopia ótica, os resultados obtidos mostraram que foi possível cultivar as CTMs com os dois microcarregadores mantendo boas condições para o crescimento tridimensional. Contudo, o rendimento celular foi limitado, principalmente, pela formação de aglomerados de microcarregadores/matriz extracelular sendo possível obter um fator de expansão celular de 4,98 com o Cytodex 1 e de 9,70 com o Cultispher S. A fácil recuperação da célula, vantagem oferecida por este microcarregador, foi aproveitada para realizar a análise da manutenção do fenótipo antigênico por citometria de fluxo e por indução da diferenciação em adipócitos e osteócitos. Os resultados comprovaram a conservação das características fenotípicas da hMSC-TERT com a técnica de cultivo avaliada neste trabalho.

Made available in DSpace on 2014-12-17T15:37:09Z (GMT). No. of bitstreams: 1 RitaCC_DISSERT.pdf: 2507687 bytes, checksum: 4eeca6582506786804976f0255257b80 (MD5) Previous issue date: 2012-03-22 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The aerial activities, leaps and slaps with parts of the body in the surface of water, are part of the behavioral repertoire of several species of cetaceans. Among them, the spinner dolphin, Stenella longirostris, shows greater diversity in such behavior. For the spinner dolphins of Fernando de Noronha, the aerial activities are classified as vertical and horizontal, with eight patterns to be noted (tail slap, head slap, motor boating, partial leap, leap, spin, tail over head and tail over head with spin) discriminated between these categories. Such behaviors can be used as a parameter to identify behavioral changes, as well as patterns of daily and seasonal activity. In this manner, this study aimed to characterize the frequency in performance of such activity while the dolphins were within the Dolphin Bay of Fernando de Noronha, and verify possible daily and seasonal hourly fluctuations on such behaviors. The data analyzed in this study was acquired during the period of January 2006 through December 2010, totaling 1431 days of observation from land set point, with 113027 aerial activities registered, daily average of 72,27 (SD=96,10). During 5478h and 54 min of observation the horizontal aerial activity was the most observed and rotation was the most executed pattern. Greater frequency of execution of aerial activity was observed in adults, but for both adults and calves, was observed a predominance of horizontal activities, with spin being the pattern most executed. Positive correlation was observed between the amount of aerial activity performed and the number of animals inside the Bay. Hourly daily fluctuation was observed in the expression of aerial activities by spinner dolphins, and was observed a peak of activity between 8h and 8h59min for the overall frequency relative of aerial activities, as well as for the categories and patterns. Seasonal differences were observed between the rainy and dry season with the greater amount of activity being observed during the rainy season. Nevertheless, the same profile of frequency relative of aerial activity was observed in both seasons with the peak amount being during the same period. When discriminated the aerial activities in categories and patterns, for both seasons, there was a similar pattern of hourly fluctuation; for most of parameters, higher frequency relative of execution of aerial activity remain between 8h and 8h59min / As atividades a?reas, saltos e batidas com partes do corpo na superf?cie da ?gua, comp?em parte do repert?rio comportamental de diversas esp?cies de cet?ceos. Dentre elas, acredita-se que o golfinho-rotador, Stenella longirostris, seja a esp?cie que apresenta maior agilidade em tais comportamentos. Para os golfinhos-rotadores de Fernando de Noronha, as atividades a?reas s?o categorizadas em atividades a?reas verticais e horizontais, sendo oito (batida de cauda, batida de cabe?a, motor de popa, ca?da, salto, rota??o, invers?o e pirueta) os padr?es discriminados entre essas classes. Tais comportamentos podem ser usados como par?metros para identificar altera??es comportamentais, bem como os padr?es de atividade di?ria e sazonais. Desta forma, o presente estudo teve por objetivo caracterizar a frequ?ncia de execu??o de atividades a?reas no decorrer do per?odo em que os golfinhos-rotadores permanecem dentro da Ba?a dos Golfinhos de Fernando de Noronha, bem como verificar a exist?ncia de poss?veis flutua??es hor?rias di?rias e sazonais na exibi??o de tais comportamentos. Para isso, foram analisados dados que compreendem o per?odo entre janeiro de 2006 a dezembro de 2010, totalizando 1.431 dias de observa??o em ponto fixo, sendo registradas 113.027 atividades a?reas, m?dia di?ria de 72,27 (DP=96,10). Nas 5.478 horas e 54 minutos de observa??o, as atividades a?reas horizontais foram observadas em maior frequ?ncia, sendo a rota??o o padr?o mais executado. Maior frequ?ncia de execu??o das atividades a?reas foi observada em indiv?duos adultos, mas tanto para adultos quanto para filhotes, observou-se predom?nio das atividades horizontais, com a rota??o sendo o padr?o mais executado. Correla??o positiva foi observada entre a quantidade de atividades a?reas executadas e o n?mero de animais dentro da Ba?a. Flutua??o hor?ria di?ria foi observada na express?o das atividades a?reas pelos golfinhos-rotadores, sendo observado maior pico de atividade no hor?rio entre 8h e 8h59min, tanto para a frequ?ncia relativa total de atividades a?reas, quanto para as classes e os padr?es. Diferen?as entre as esta??es clim?ticas seca e chuvosa foram observadas, com maior registro de atividade a?rea durante o per?odo de chuvas. Entretanto, o perfil de distribui??o da frequ?ncia relativa de atividades a?reas se assemelha para ambas as esta??es, mantendo o hor?rio de maior pico de atividade. Quando discriminadas as atividades a?reas nas classes e nos padr?es considerados, para ambas as esta??es observaram-se padr?o de flutua??o hor?ria semelhante; para a maioria dos par?metros analisados, maior frequ?ncia relativa de execu??o das atividades a?reas se mant?m entre 8h e 8h59min

Células de mamíferos são os hospedeiros mais frequentemente utilizados para a fabricação de proteínas biofarmacêuticas e para a produção de vacinas virais, A qualidade é um elemento-chave para o estabelecimento de um processo de bioconversão eficiente. No presente trabalho utilizamos a linhagem de células BHK- 21C13(Baby Hamster Kidney) adaptadas para cultivo em suspensão. O uso de Soro Fetal Bovino (SFB) é tradicionalmente utilizado, sendo considerado um suplemento universal, pois permite o crescimento em várias linhagens de células de mamíferos; porém, uso de SFB apresenta risco de infecção por prions, variabilidade entre lotes e aumento no custo em etapa de purificação (Downstream processing). O objetivo do presente trabalho foi comparar o cultivo de células BHK-21 C13 entre dois meios suplementados com SFB e sem SFB, através do estudo cinético para cultivo em suspensão estático e agitado com frascoT, frasco spinner e biorreator, respectivamente. Os parâmetros; Xmáx e &#181;máx, não foram significativamente influenciados pelo meio de cultura em cultivo estático, em cultivo com agitação em frasco spinner e também no cultivo em biorreator. O tempo de duplicação ficou próximo para todas as condições testadas. A produtividade alcançada foi: 0,032x106 cel/mL.h-1 para o meio com SFB e 0,031 X106 cel/mL.h-1 para o meio sem SFB. Ao final do processo foi possível obter uma concentração celular em torno de 4,7x106 cel/mL, tanto para o cultivo com SFB quanto para o cultivo sem SFB. Dessa forma, o uso de meio de cultivo sem SFB não alterou os principais parâmetros cinéticos, não apresentando as desvantagens do uso do SFB. / Mammalian cells are the most frequently used hosts for the production of biopharmaceutical proteins and viral vaccines. Quality is a key element for the establishment of an efficient bioconversion process. In this work, we used the cell line Baby Hamster Kidney C13 (BHK-21 C13) adapted to suspension culture was used. Fetal Bovine Serum (FBS) is traditionally used and it is considered a universal insert due to its power to increase cell growth in this kind of animal cells. However, the utilization of FBS introduces risks of infection from prions, variability between batches and increase in cost associated to purification stages (downstream processing). This work aimed to compare the kinetic behaviors of BHK-21 C13 cells in two media supplemented with FBS and without FBS using both one static and two suspension systems, T-flask, spinner flask and bioreactor respectively. The parameters; Xmax and &#181;max were not significantly influenced by the culture medium in T- flask culture static, in spinner flask cultivation and were neither significantly influenced by growing in culture media stirred bioreactor. The doubling time was close to all conditions tested. At the end of the growth phase it was possible to obtain a nearby cell concentration of 4.7 x 106 cells / ml, both for cultivation with FBS as for FBS without cultivation. Thus, the use of culture medium without FBS did not affect the main kinetic parameters. Besides, it does not show the disadvantages of culture media using FBS.