Cell migration is a fundamental biological process involved in tissue development, wound repair, and diseases such as cancer metastasis. It is a biomechanical process involving the adhesion of a cell to a substratum, usually an elastic extracellular matrix, as well as the physical contraction of the cell driven by intracellular actomyosin network. In the migration of cells as a group, known as collective migration, the cells are also physically linked to one another through cell-cell adhesions. How mechanical interactions with cell substratum and with neighboring cells regulate movements during collective migration, nevertheless, is poorly understood. To address this question, the effects of substrate stiffness on sheet migration of MCF10A epithelial cells were systematically analyzed. Speed, persistence, directionality and coordination of individual cells within the migrating sheet were all found to increase with substrate stiffening. Substrate stiffening also enhanced the propagation of coordinated movement from the sheet edge into the monolayer, which correlated with an upregulation of myosin-II activity in sheet edge cells. This mechano-response was dependent on cadherin-mediated cell-cell adhesions, which are required for the transmission of directional cue. Importantly, myosin-II contractility modulated cadherin- dependent cell-cell coordination, suggesting that contractile forces at cadherin adhesions regulate collective migration. To measure forces transmitted through cell-cell adhesions, a quantitative approach was developed in which cell-cell forces were deduced from cell-substrate traction forces, based on force balance principles and simple cell mechanics modeling. This method enabled the analysis of cell-cell mechanical interactions in small cell clusters of complex topology. The dynamic fluctuations of cell-cell forces over time revealed that force transmission between non-adjacent cells is typically limited, but is enhanced when the cell across which forces are being transmitted has reduced myosin-IIA or talin-1. This suggests that cells in a group may differentially regulate their levels of myosin-II contractility and cell-matrix mechanotransduction to promote longer-range force transmission during collective migration. Together, the results in this dissertation led to a working model of collective cell migration as regulated by cell-matrix mechanical properties and cell-cell mechanical interactions. This model, as well as the quantitative techniques developed here, will drive future studies on the mechanisms underlying collective migration.

Living organisms are made up of a multitude of individual cells that are surrounded by biomolecules and fluids. It is well known that cells are highly regulated by biochemical signals; however it is now becoming clear that cells are also influenced by the mechanical forces and mechanical properties of the local microenvironment. Extracellular forces causing cellular deformation can originate from many sources, such as fluid shear stresses arising from interstitial or blood flow, mechanical stretching during breathing or compression during muscle contraction. Cells are able to sense variations in the mechanical properties (elasticity) of their microenvironment by actively probing their surroundings by utilizing specialized proteins that are involved in sensing and transmitting mechanical information. The actin cytoskeleton and myosin-II motor proteins form a contractile (actomyosin) network inside the cell that is connected to the extracellular microenvironment through focal adhesion and integrin sites. The transmission of internal actomyosin strain to the microenvironment via focal adhesion sites generates mechanical traction forces. Importantly, cells generate traction forces in response to extracellular forces and also to actively probe the elasticity of the microenvironment. Many studies have demonstrated that extracellular forces can lead to rapid cytoskeletal remodeling, focal adhesion regulation, and intracellular signalling which can alter traction force dynamics. As well, cell migration, proliferation and stem cell fate are regulated by the ability of cells to sense the elasticity of their microenvironment through the generation of traction forces. In vitro studies have largely explored the influence of substrate elasticity and extracellular forces in isolation, however, in vivo cells are exposed to both mechanical cues simultaneously and their combined effect remains largely unexplored. Therefore, a series of experiments were performed in which cells were subjected to controlled extracellular forces as on substrates of increasing elasticity. The cellular response was quantified by measuring the resulting traction force magnitude dynamics. Two cell types were shown to increase their traction forces in response to extracellular forces only on substrates of specific elasticities. Therefore, cellular traction forces are regulated by an ability to sense and integrate at least two pieces of mechanical information - elasticity and deformation. Finally, this ability is shown to be dependent on the microtubule network and regulators of myosin-II activity.

SILVIA, Silvia Ferreira da. Avaliação do ruído e do desempenho energético de um trator de rabiças com um arado de disco acoplado. 2012. 73 f. Dissertação (Mestrado em engenharia agrícola)- Universidade Federal do Ceará, Fortaleza-CE, 2012. / Submitted by Elineudson Ribeiro (elineudsonr@gmail.com) on 2016-06-28T19:03:33Z No. of bitstreams: 1 2012_dis_sfsilva.pdf: 2169759 bytes, checksum: c7992e3e903a610067c88893f4c68971 (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-06-30T22:51:17Z (GMT) No. of bitstreams: 1 2012_dis_sfsilva.pdf: 2169759 bytes, checksum: c7992e3e903a610067c88893f4c68971 (MD5) / Made available in DSpace on 2016-06-30T22:51:17Z (GMT). No. of bitstreams: 1 2012_dis_sfsilva.pdf: 2169759 bytes, checksum: c7992e3e903a610067c88893f4c68971 (MD5) Previous issue date: 2012 / The initial tillage has great importance for agricultural production. This activity is usually performed by animal traction in family farming. In this paper, we studied the use of handlebar tractors as a source for field work optimization. The experiment was conducted with a handlebar tractor with a plow attached to it at the Federal University of Ceará. The goal was to evaluate the energetic efficiency of the handlebar traitor attached to the disc plow, using three gears (1st, 2nd, 3rd) in a red, yellow argisoil. It was also evaluated the soil shoaling and the noise levels with the machine both stopped in different rays of distances (1, 2, 5 and 10 meters), and with handlebar traitor in movement, using three different number of turns (810, 1040 and 1360 rpm). To obtain data about energetic efficiency, it was used a handlebar traitor, an instrument with inductive sensors on wheels to measure the wheelsets slippage, a flowmeter to measure the fuel consumption and a loading cell to obtain the workforce in the drawbar. To install the loading cell and obtain the implement stability, it was necessary to use a train, in which the set traveled 25m with three repetitions in each gear to the collecting of the data. The soil shoaling was calculated through the data obtained in the field by a profilometer, and the levels of noise were measured with a dosimeter and a decibelimeter. The studied variables were the hourly fuel consumption (L.h-1), the specific fuel consumption, (g.kW.h-1), the wheelsets slippage (%), traction force in the bar (kN), work speed (m.s-1) and available power (kW). These data were evaluated with the plow working and with the implement raised to be compared. We calculated the difference (results with the implement plowing minus the results with the plow not working) to study the obtained averages. Data analysis was performed using descriptive statistics using the computer program Assistat 7.6 beta. The results showed that the average rates of slippage were 10% (1st gear), 12 % (2nd gear) and 18% (3rd gear). The average rates for the plow while it was working for the hourly fuel consumption were 5,45 L.h-1 (1st gear), 4,22 L.h-1 (2nd gear) and 2,54 L.h-1 (3rd gear); the specific fuel consumption was 3004,98 g.kW.h-1 (1st gear), 4343,88 g.kW.h-1 (2nd gear) and 5787,91 g.kW.h-1 (3rd gear); the power was 1,54; 0,83 and 0,38 kW for 1st, 2nd and 3rd gear respectively, and the movement speed averages were 0,32 m.s-1 (1st gear), 0,41 m.s-1 (2nd gear) and 0,44 m.s-1 (3rd gear). The average rates for the hourly fuel consumption that were found for the moment when the plow was not working, that is, the implement was raised, were 6,58 L.h-1 (1st gear), 5,98 L.h-1 (2nd gear) and 4,20 L.h-1 (3rd gear); the rates for the specific fuel consumption were 3514,09 g.kW.h-1 (1st gear), 4728,22 g.kW.h-1 (2nd gear) and 5787,91 g.kW.h-1 (3rd gear); the power was 1,54 kW (1st gear), 0,83 kW (2nd gear) and 0,38 kW (3rd gear), and the movement speed averages were 0,27; 0,33 and 0,40 m.s-1 for 1st , 2nd and 3rd gears respectively. The soil shoaling achieved the rates 75 % (2nd gear) and, approximately, 64 % for the 1st and 3rd gears. And the levels of noise that were found by the usage of the dosimeter were 95,7dB(A) (1st gear), 98,2dB(A) (2nd gear) and 98,8 dB(A) (3rd gear). The average levels of noise that were found by the usage of the decibelimeter were 76,71; 77,38 and 80,80 dB(A) for the turns 810, 1.040 e 1.360 rpm respectively. It is advised the use of headphones or the decreasing of the working hours for the operator who is working directly with the tractor handlebars. / O preparo inicial do solo tem grande importância para a produção agrícola, essa atividade é geralmente realizada por tração animal na agricultura familiar. Nesse trabalho estudou-se a utilização de tratores de rabiças como fonte de otimização do trabalho no campo. O experimento foi desenvolvido com um trator de rabiças com um arado acoplado na Universidade Federal do Ceará com o objetivo de avaliar a eficiência energética do trator de rabiças acoplado ao arado de disco utilizando três marchas (1ª, 2ª e 3ª) em um Argissolo Vermelho Amarelo, avaliou-se também o empolamento do solo e os níveis de ruído com a máquina parada em diferentes raios de afastamento (1, 2, 5 e 10 m), e com o trator de rabiças em movimento utilizando três rotações (810, 1.040 e 1.360 rpm). Para obtenção de dados sobre a eficiência energética utilizou-se um trator de rabiças, instrumentado com sensores indutivos nas rodas para medir a patinagem dos rodados, fluxômetro para medida do consumo de combustível e célula de carga para obtenção da força de trabalho na barra de tração. Para instalar a célula de carga e obter estabilidade do implemento foi necessário utilizar um comboio, onde o conjunto percorria um espaço 25 m com três repetições em cada marcha para as coletas dos dados. O empolamento do solo foi calculado através de dados obtidos com o perfilometro no campo e os níveis de ruído foram medidos com dosímetro e decibelimetro. As variáveis estudadas foram consumo horário de combustível (L.h-1), consumo específico de combustível (g.kW.h-1), patinagem dos rodados (%), força de tração na barra (kN), velocidade de trabalho (m.s-1) e potência disponível (kW). Esses dados foram avaliados com o arado realizando trabalho e com o implemento erguido para ser feito de comparação calculou-se as diferença (resultados com o implemento arando, menos os resultados com o arado sem realizar trabalho), para estudo das médias obtidas. A análise dos dados foi realizada utilizando estatística descritiva utilizando o programa computacional Assistat 7.6 beta. Os resultados mostraram que os valores médios da patinagem foram de 10 % (1ª marcha), 12 % (2ª marcha) e 18 % (3ª marcha). Os valores médios encontrados para o arado realizando trabalho para o consumo horário de combustível foi de 5,45 L.h-1 (1ª marcha), 4,22 L.h-1 (2ª marcha) e 2,54 L.h-1 (3ª marcha); o consumo específico de combustível foi de 3004,98 g.kW.h-1 (1ª marcha), 4343,88 g.kW.h-1 (2ª marcha) e 5787,91 g.kW.h-1 (3ª marcha); a potência foi de 1,54; 0,83 e 0,38 kW para a 1ª, 2ª e 3ª marcha respectivamente e as médias da velocidade de deslocamento foram de 0,32 m.s-1 (1ª marcha), 0,41 m.s-1 (2ª marcha) e 0,44 m.s-1 (3ª marcha). Os valores médios encontrados para o momento em que o arado não realizava trabalho, ou seja, o implemento estava erguido foi para o consumo horário de combustível de 6,58 L.h-1 (1ª marcha), 5,98 L.h-1 (2ª marcha) e 4,20 L.h-1 (3ª marcha); o consumo específico de combustível apresentou valores de 3514,09 g.kW.h-1 (1ª marcha), 4728,22 g.kW.h-1 (2ª marcha) e 5787,91 g.kW.h-1 (3ª marcha); a potência foi de 1,54 kW (1ª marcha), 0,83 kW (2ª marcha) e 0,38 kW (3ª marcha) e as médias da velocidade de deslocamento foram de 0,27; 0,33 e 0,40 m.s-1 para a 1ª, 2ª e 3ª marchas respectivamente. O empolamento do solo chegou a 75 % (2ª marcha) e obteve valores de aproximadamente 64 % para a 1ª e 3ª marcha. E os níveis de ruído encontrados utilizando o dosimetro foram de 95,7 dB(A) (1ª marcha), de 98,2 dB(A) (2ª marcha) e 98,8 dB(A) (3ª marcha). Os níveis médios de ruído encontrados com o auxilio do decibelimetro foi de 76,71; 77,38 e 80,80 dB(A) para as rotações de 810, 1.040 e 1.360 rpm respectivamente. Recomenda-se o uso de protetores auriculares ou a diminuição da carga horária de trabalho para o operador que esteja trabalhando diretamente com o trator de rabiças.

Made available in DSpace on 2014-08-20T13:25:41Z (GMT). No. of bitstreams: 1 TESE_Mauro_Fernando_Ferreira.pdf: 34428971 bytes, checksum: 24ff7e5e9163d78b1220c1bbf56e782c (MD5) Previous issue date: 2008-11-28 / This work had as objective to analyze alternatives to increase the useful life of the ferrules of hoe opener rods of no-till planters, through the addition of covering welding. It was verified the performance of five welding types, being used covered electrodes and tubular wire, over the extremity of the hoe. The tests were executed in franc-sandy soil characterized physically by its texture, plasticity index, density, and water tenor, as well as the resistance to the penetration. The wearing, the horizontal and vertical force were obtained in intervals of 10,5km to the limit of 52,5km. The results indicated that the witness ferrule has lost in the average of the experiment 38,41% of its initial mass when it traveled 52,5km while the other welded ferrules traveled between 61,0km and 108,7km to reach the same mass reductions. It was observed that the vertical and horizontal forces contributed with 11% and 89%, respectively, of the resulting force in the hoe. With the use of the coverings, it was obtained an increase of the area worked in the sowing, for the largest durability of the tools, between 2,0ha and 13,4ha, in relation to the witness. The theoretical field efficiency was larger when it was used the coverings. The tubular wire increased in 5,1% in relation to the ferrule without covering. Starting from the found results we can conclude that it is possible to increase the useful life of the ferrules of furrow opener rods through the addition of welding material and that the conservation of the geometry of the tools, provides the maintenance of the forces that have action on these ones as well as their functional characteristics. / Este trabalho teve como objetivo analisar alternativas para o aumento da vida útil das ponteiras de hastes sulcadoras de semeadoras adubadoras, através da adição de soldagem de revestimento. Verificou-se o desempenho de cinco tipos de soldagem, utilizando-se eletrodos revestidos e arame tubular, sobre a extremidade das ponteiras. Os testes foram executados em solo franco-arenoso caracterizado fisicamente pela sua textura, índice de plasticidade, densidade e teor de água, assim como a resistência à penetração. O desgaste, a força horizontal e vertical foram obtidos em intervalos de 10,5km até o limite de 52,5km. Os resultados indicaram que a ponteira testemunha perdeu na média do experimento 38,41% da sua massa inicial quando percorreu 52,5km enquanto que as demais ponteiras soldadas percorreram entre 61,0km e 108,7km para atingir as mesmas reduções de massa. Observou-se que as forças verticais e horizontais contribuíram com 11% e 89%, respectivamente, da força resultante na ponteira. Com o uso dos revestimentos obteve-se um aumento da área trabalhada na semeadura, pela maior durabilidade das ferramentas, entre 2,0ha e 13,4ha, em relação à testemunha. A eficiência de campo teórica foi maior quando se utilizou os revestimentos sendo que o arame tubular aumentou em 5,1% em relação à ponteira sem revestimento. A partir dos resultados encontrados conclui-se que se pode aumentar a vida útil das ponteiras de hastes sulcadoras através da adição de material soldável e que a conservação da geometria das ferramentas, proporciona a manutenção das forças atuantes sobre estas assim como a suas características funcionais.

Les cellules épithéliales forment des tissus cohésifs, sous forme de monocouches que l'on retrouve dans les poumons, les reins ou la peau. Travaillant à partir d'expériences in vitro, nous avons caractérisé le comportement mécanique de monocouches cellulaires. Nous avons étudié la fermeture de blessures circulaires sur un substrat non adhésif. En comparant différents modèles, nous avons montré comment la fermeture est possible grâce à un cable contractile d'acto-myosine et aux fluctuations de la tension du tissu. La Microscopie des Forces de Traction (TFM) permet de mesurer les forces que les cellules exercent sur leur substrat. A partir de cette mesure et en utilisant l'équilibre des forces, nous avons développé une méthode qui résout ce problème sous-déterminé par inversion bayésienne et permet d'obtenir le champ des contraintes internes au tissu. En appliquant cette méthode sur des images (BISM: Microscopie des contraintes par inversion bayésienne) et en l'adaptant à l'aide d'un filtre de Kalman sur des films (KISM: Microscopie des contraintes par inversion de Kalman), nous avons inféré le tenseur des contraintes de monocouches cellulaires sans faire aucune hypothèse sur la rhéologie du tissu. Enfin, nous avons estimé les contraintes directement depuis les déplacements du substrat, sans passer par les forces de tractions et donc en réduisant le nombre d'inversions de matrice (BISMu: Microscopie des contraintes par inversion bayésienne à partir des déplacements du substrat). / Epithelial cells are known to form cohesive monolayers, a form of tissue organization encountered in the lung, the kidney or the skin. From in vitro experiments, we have characterized the mechanical properties of cell monolayers. We have studied the closure of circular wounds over a nonadhesive substrate. Comparing different models, we have shown how closure is possible thanks to a contractile acto-myosin cable and to fluctuations of the tissue tension. Traction Force Microscopy (TFM) allows to measure the forces that cells exert on their substrate. Starting from this measurement and using the force balance equations, we have solved this underdetermined problem by Bayesian inversion and obtained the internal stress field of the tissue. Applying this method on single images (BISM: Bayesian Inversion Stress Microscopy), and adapting it with a Kalman filter for movies (KISM: Kalman Inversion Stress Microscopy) we have inferred the stress tensor of cell monolayers, without making any hypothesis on the tissue rheology. Finally, we have estimated the stresses directly from the substrate displacements, without computing the traction forces and thus reducing the number of matrix inversions (BISMu: Bayesian Inversion Stress Microscopy from substrate displacements).

Living organisms are made up of a multitude of individual cells that are surrounded by biomolecules and fluids. It is well known that cells are highly regulated by biochemical signals; however it is now becoming clear that cells are also influenced by the mechanical forces and mechanical properties of the local microenvironment. Extracellular forces causing cellular deformation can originate from many sources, such as fluid shear stresses arising from interstitial or blood flow, mechanical stretching during breathing or compression during muscle contraction. Cells are able to sense variations in the mechanical properties (elasticity) of their microenvironment by actively probing their surroundings by utilizing specialized proteins that are involved in sensing and transmitting mechanical information. The actin cytoskeleton and myosin-II motor proteins form a contractile (actomyosin) network inside the cell that is connected to the extracellular microenvironment through focal adhesion and integrin sites. The transmission of internal actomyosin strain to the microenvironment via focal adhesion sites generates mechanical traction forces. Importantly, cells generate traction forces in response to extracellular forces and also to actively probe the elasticity of the microenvironment. Many studies have demonstrated that extracellular forces can lead to rapid cytoskeletal remodeling, focal adhesion regulation, and intracellular signalling which can alter traction force dynamics. As well, cell migration, proliferation and stem cell fate are regulated by the ability of cells to sense the elasticity of their microenvironment through the generation of traction forces. In vitro studies have largely explored the influence of substrate elasticity and extracellular forces in isolation, however, in vivo cells are exposed to both mechanical cues simultaneously and their combined effect remains largely unexplored. Therefore, a series of experiments were performed in which cells were subjected to controlled extracellular forces as on substrates of increasing elasticity. The cellular response was quantified by measuring the resulting traction force magnitude dynamics. Two cell types were shown to increase their traction forces in response to extracellular forces only on substrates of specific elasticities. Therefore, cellular traction forces are regulated by an ability to sense and integrate at least two pieces of mechanical information - elasticity and deformation. Finally, this ability is shown to be dependent on the microtubule network and regulators of myosin-II activity.

No description available.

This study presents a novel approach for predicting future cellular traction in a time series. The proposed method leverages two distinct look-ahead Long Short-Term Memory (LSTM) models—one for cell boundary and the other for traction data—to achieve rapid and accurate predictions. These LSTM models are trained using real Fourier Transform Traction Cytometry (FTTC) output data, ensuring consistency and reliability in the underlying calculations. To account for variability among cells, each cell is trained separately, mitigating generalized errors. The predictive performance is demonstrated by accurately forecasting tractions for the next 30-time instances, with an error rate below 7%. Moreover, a strategy for real-time traction calculations is proposed, involving the capture of a bead reference image before cell placement in a controlled environment. By doing so, we eliminate the need for cell removal and enable real-time calculation of tractions. Combining these two ideas, our tool speeds up the traction calculations 1.6 times, leveraging from limiting TFM use. As a walk forward, prediction method is implemented by combining prediction values with real data for future prediction, it is indicative of more speedup. The predictive capabilities of this approach offer valuable insights, with potential applications in identifying cancerous cells based on their traction behavior over time.Additionally, we present an advanced cell boundary detection algorithm that autonomously identifies cell boundaries from obscure cell images, reducing human intervention and bias. This algorithm significantly streamlines data collection, enhancing the efficiency and accuracy of our methodology.

Yes / Cell migration is a key contributor to wound repair. This study presents findings indicating that the liquid crystal based cell traction force transducer (LCTFT) system can be used in conjunction with a bespoke cell traction force mapping (CTFM) software to monitor cell/surface traction forces from quiescent state in real time. In this study, time-lapse photo microscopy allowed cell induced deformations in liquid crystal coated substrates to be monitored and analyzed. The results indicated that the system could be used to monitor the generation of cell/surface forces in an initially quiescent cell, as it migrated over the culture substrate, via multiple points of contact between the cell and the surface. Future application of this system is the real-time assaying of the pharmacological effects of cytokines on the mechanics of cell migration.

Phagocytosis is the process by which cells engulf foreign bodies. It is the hallmark behavior of white blood cells, being the process through which those cells ingest and degrade pathogens and debris. To date a large amount of research has focused on documenting the existence and role of biochemical components involved with phagocytosis. Scores of signaling molecules have been implicated in the complex signal cascade which drives the process. These molecules are small (typically no larger than 5 nanometers) and operate in a crowded, chemically “noisy,” environment, yet they coordinate the cell's activity over comparatively expansive distances (as large as 20 micrometers). How these molecular processes scale-up to coordinate the activities of the cell over such massive distances is largely unknown. Using a planar analog of phagocytosis termed “frustrated phagocytosis,” we experimentally demonstrate that phagocytosis occurs in three distinct phases: initial cell-antigen binding, symmetric spreading, and late-stage contraction. Initial binding and symmetric spreading appears to be both mechanically and chemically similar to the quasi-universal cellular behaviors of adhesion and migration. Adhesion and migration have received extensive attention from the biophysics community in recent years. Leveraging these similarities, we adapt the biomechanical frameworks used in models of migration to phagocytosis. We show that macroscopic properties such as a cell's effective viscosity and membrane cortical tension can be used to model cell behavior during phagocytosis. Our experiments reveal that late-stage contraction distinguishes frustrated phagocytosis from other spreading behaviors. This contraction is myosin dependent. Additionally we demonstrate, for the first time, that late-stage contraction corresponds with formation of a contractile F-actin belt. Based on the dynamic contraction model (DC) developed to explain actin structure during cell migration we propose a DC model of phagocytosis which posits that contractile belt formation is the result of a late-stage myosin activity coupled with F-actin.