This action research project examines the role classroom culture and discourse can play on student learning, with a focus on female students. A sixth grade science classroom was evaluated through analysis of two videotaped astronomy lessons. The classroom environment utilized qualitative methods to examine teacher and student interactions, student and student interactions, and classroom environment. The research project began in response to a previous research project which found that after completing an astronomy unit male students not only out preformed female students, but female students lost gains in several area. Findings suggested that there may be a connection between the classroom discourse and microculture and the girls’ low performance.

The goal of this study is to design a protocol for the adherent cell culture within a novel microfluidic device. Microscale cell culture protocols were developed for loading cells using poly-L-lysine to enhance adherent cell culture of murine derived NIH 3T3 fibroblasts. This work sought to develop a method for adherent microculture by examining various sterilization, surface treatment, and seeding techniques. Using a vacuum suction loading technique, air plasma treatment and a poly-L-lysine surface treatment adherent cell culture was observed within the device. The work presented here is part of a collaborative effort that aims to develop protocols for the electrical and optical characterization of cell culture within a novel microfluidic device.

<p>ABSTRACT</p><p>Chemotherapy is a common method used for cancer treatment. Especially when it concerns cancers that have grown invasively it seems to be the only efficient treatment due to the substances ability to reach and affect almost the entire body. One major obstacle regarding chemotherapy is that the patients often develop resistance to the cytotoxic substances used. Fluorometric microculture cytotoxicity assay (FMCA) is a method developed to measure sensitivity of tumor cells to different cytotoxic substances in vitro. The assay is based on hydrolysis of fluorescein diacetate to fluorescein by cells with intact cell membranes after incubation with drugs for 72 hours. This study investigated the impact of two methodological factors that may cause errors in the achieved results; namely the possible occurrence of drug decay during incubation and the use of an inappropriate amount of cells. These factors were tested by exposing the cytotoxic drugs to pre-incubation in absence of tumor cells for different times and to use suspensions with different concentrations of cells. The results indicated occurrence of drug decay in 3 of the 18 substances tested and that the amount of cells affected the results for most of the drugs tested but to different extent.</p>

No esporte podemos identificar microculturas constituídas por um grupo de pessoas que compartilham valores, crenças, padrões de comportamentos e um sistema de símbolos, verbal e não verbal, que os distingue do meio cultural dominante. O ginásio é o espaço no qual as microculturas de Ginástica Artística (GA) estão resguardadas e mantém a preeminência das tradições que constituem, em âmbito maior, a macrocultura da modalidade. Nesse ambiente há uma teia de significados, supostamente compartilhados, que emergem de suas estruturas de ação, comportamento e comunicação que apresentam características idiossincráticas. E, essas só podem ser compreendidas a partir dos significados atribuídos pelos seus próprios protagonistas, ou seja, ginastas e técnicos. O objetivo do presente estudo foi identificar, apresentar e analisar aspectos que constituem a microcultura de GA, em um ginásio de treinamento de alto rendimento, na categoria feminina. O caminho metodológico escolhido foi a abordagem de um estudo de caso do tipo etnográfico. No transcorrer do estudo, foi possível desvelar procedimentos, valores e comportamentos que elucidam a cultura de treinamento da GA feminina e que conformam a dinâmica de funcionamento do ginásio. Observamos no corpo e no desempenho das ginastas aspectos que refletem a cultura, assim como nos padrões de comportamento e na comunicação das ginastas e dos técnicos. Valores como a perseverança, a disciplina, a dedicação e a subserviência das ginastas aos técnicos emergiram durante as horas de treinamento. Ademais, observamos atributos inerentes ao universo simbólico do ginásio como a necessidade de sacrifício à dor e os atos ritualísticos. Essa cultura de treinamento, meio pelo qual as ginastas e os técnicos se adaptam e fornecem sentido às suas ações nesse ambiente físico e social de treinamento, expôs características peculiares da GA feminina e que continuam a ser transmitidas entre as gerações de atletas pelo processo de endoculturação / In the sport environment we can identify microcultures consisting of a group of people who share values, beliefs, patterns of behavior and a symbolic system, verbal and nonverbal, which distinguishes them from the dominant cultural milieu. The gym is the space in which Artistic Gymnastics microcultures are sheltered and preserves the preeminent traditions which constitute, at a larger scope, the macroculture of this sport. In this environment there is a web of meanings, supposedly shared, emerging from their structures of action, behavior and communication that have idiosyncratic characteristics. Moreover, that can only be understood from the meanings assigned by its own protagonists, in other words, gymnasts and coaches. The aim of this study was to identify, present and analyze aspects that constitute the microculture of Artistic Gymnastics at a high performance training gym in the woman\'s category. The methodological path chosen consisted of a case study of ethnographic type. In the course of the study, it was possible to unveil procedures, values, and behaviors that elucidated the Woman\'s Artistic Gymnastics training culture that configured the dynamic operation of the gym. We observed in the body and in the performance of the gymnasts aspects that reflect the gym microculture, as well as in patterns of behavior and communication of the gymnasts and coaches. Values such as perseverance, discipline, dedication and subservience of the gymnasts to their coaches emerged during training hours. Furthermore, we observed inherent attributes emerging from the gym symbolic universe such as the need of pain sacrifice and ritualistic acts. This training culture, the means by which the gymnasts and coaches adapt themselves and provide significance to their actions in the physical and social environment of training, showed peculiar characteristics of Woman\'s Artistic Gymnastics that continue to be passed down across the generation of athletes by endoculturization process

No esporte podemos identificar microculturas constituídas por um grupo de pessoas que compartilham valores, crenças, padrões de comportamentos e um sistema de símbolos, verbal e não verbal, que os distingue do meio cultural dominante. O ginásio é o espaço no qual as microculturas de Ginástica Artística (GA) estão resguardadas e mantém a preeminência das tradições que constituem, em âmbito maior, a macrocultura da modalidade. Nesse ambiente há uma teia de significados, supostamente compartilhados, que emergem de suas estruturas de ação, comportamento e comunicação que apresentam características idiossincráticas. E, essas só podem ser compreendidas a partir dos significados atribuídos pelos seus próprios protagonistas, ou seja, ginastas e técnicos. O objetivo do presente estudo foi identificar, apresentar e analisar aspectos que constituem a microcultura de GA, em um ginásio de treinamento de alto rendimento, na categoria feminina. O caminho metodológico escolhido foi a abordagem de um estudo de caso do tipo etnográfico. No transcorrer do estudo, foi possível desvelar procedimentos, valores e comportamentos que elucidam a cultura de treinamento da GA feminina e que conformam a dinâmica de funcionamento do ginásio. Observamos no corpo e no desempenho das ginastas aspectos que refletem a cultura, assim como nos padrões de comportamento e na comunicação das ginastas e dos técnicos. Valores como a perseverança, a disciplina, a dedicação e a subserviência das ginastas aos técnicos emergiram durante as horas de treinamento. Ademais, observamos atributos inerentes ao universo simbólico do ginásio como a necessidade de sacrifício à dor e os atos ritualísticos. Essa cultura de treinamento, meio pelo qual as ginastas e os técnicos se adaptam e fornecem sentido às suas ações nesse ambiente físico e social de treinamento, expôs características peculiares da GA feminina e que continuam a ser transmitidas entre as gerações de atletas pelo processo de endoculturação / In the sport environment we can identify microcultures consisting of a group of people who share values, beliefs, patterns of behavior and a symbolic system, verbal and nonverbal, which distinguishes them from the dominant cultural milieu. The gym is the space in which Artistic Gymnastics microcultures are sheltered and preserves the preeminent traditions which constitute, at a larger scope, the macroculture of this sport. In this environment there is a web of meanings, supposedly shared, emerging from their structures of action, behavior and communication that have idiosyncratic characteristics. Moreover, that can only be understood from the meanings assigned by its own protagonists, in other words, gymnasts and coaches. The aim of this study was to identify, present and analyze aspects that constitute the microculture of Artistic Gymnastics at a high performance training gym in the woman\'s category. The methodological path chosen consisted of a case study of ethnographic type. In the course of the study, it was possible to unveil procedures, values, and behaviors that elucidated the Woman\'s Artistic Gymnastics training culture that configured the dynamic operation of the gym. We observed in the body and in the performance of the gymnasts aspects that reflect the gym microculture, as well as in patterns of behavior and communication of the gymnasts and coaches. Values such as perseverance, discipline, dedication and subservience of the gymnasts to their coaches emerged during training hours. Furthermore, we observed inherent attributes emerging from the gym symbolic universe such as the need of pain sacrifice and ritualistic acts. This training culture, the means by which the gymnasts and coaches adapt themselves and provide significance to their actions in the physical and social environment of training, showed peculiar characteristics of Woman\'s Artistic Gymnastics that continue to be passed down across the generation of athletes by endoculturization process

ABSTRACT Chemotherapy is a common method used for cancer treatment. Especially when it concerns cancers that have grown invasively it seems to be the only efficient treatment due to the substances ability to reach and affect almost the entire body. One major obstacle regarding chemotherapy is that the patients often develop resistance to the cytotoxic substances used. Fluorometric microculture cytotoxicity assay (FMCA) is a method developed to measure sensitivity of tumor cells to different cytotoxic substances in vitro. The assay is based on hydrolysis of fluorescein diacetate to fluorescein by cells with intact cell membranes after incubation with drugs for 72 hours. This study investigated the impact of two methodological factors that may cause errors in the achieved results; namely the possible occurrence of drug decay during incubation and the use of an inappropriate amount of cells. These factors were tested by exposing the cytotoxic drugs to pre-incubation in absence of tumor cells for different times and to use suspensions with different concentrations of cells. The results indicated occurrence of drug decay in 3 of the 18 substances tested and that the amount of cells affected the results for most of the drugs tested but to different extent.

Made available in DSpace on 2016-04-29T13:18:18Z (GMT). No. of bitstreams: 1 Mariana Cavalcante Vieira.pdf: 1576858 bytes, checksum: a83e1d1db29f1a44bbcb3e8d8910ab78 (MD5) Previous issue date: 2010-04-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Metacontingency is the unit of analysis at the cultural level proposed in analogy to the three-term contingency to explain social phenomena that involves the functional relation between interlocking behavioral contingencies (IBC) and its aggregate outcome and the cultural consequence. So far, experimental research has investigated the relation equivalent to the response-reinforcer in the operant contingency. In this study, the following questions were made: (1) An antecedent stimulus similar to the DS assume evocative function to IBC and their aggregate outcome in a metacontingency? (2) What are the effects of alternating two antecedent stimuli, each one correlated to a specific metacontingency? (3) Will the establishment of an analogous to discriminative stimuli control produce an analog of generalization process? Fifteen college students participated in this study, in a total of 12 generations (3 participants in each). This study had seven experimental conditions in an trial procedure. One to three participants worked simultaneously, each one in a computer: the participant of the left (PL), center (PC) and right (PR). In each trial, the computer screen of each participant, presented four numbers, independently, in each column. The participant had to enter another four in matching columns. If the sums of the numbers presented by the computers and the numbers in each column selected by the participant resulted in odd numbers, the participant earned points. This contingency was called individual contingency. Two other contingencies focused on relations between the products of the behavior of the participants, called metacontingencies. In metacontingency 1, the background color of the screen was blue (SM1) and the sum of 4 numbers entered by the participant PL was smaller than the sum of 4 numbers entered by the participant PC and this sum was smaller than the sum of 4 numbers entered by PR (ΣPL <ΣPC <ΣPR), a cultural consequence was produced: the participants received additional credits called bonus. In metacontingency 2, the screens had a red background (SM2) and the participants produced bonus if "ΣPL> ΣPC> ΣPR . These metacontingencies were presented in random order between trials. When stability criteria was reached, older participants were replaced by newer participants. Tests of stimulus control and generalization were presented at established moments. The results showed the selection of operant behavior and metacontingencies 1 and 2 and indicated that SM1 and SM2 acquired evocative function of the corresponding metacontingencies. The tests suggested that the background colors of the screen became the stimulus dimension that exerted control over the behavior of the participants and their interactions. The data are discussed in terms of an analogy between operant contingency and metacontingency / A metacontingência é a unidade de análise no nível cultural proposta em analogia à tríplice contingência para explicar fenômenos sociais que envolvem a relação funcional entre contingências comportamentais entrelaçadas (CCE) e seu produto agregado e uma conseqüência cultural. Até o momento, as pesquisas experimentais investigaram o equivalente à relação resposta-reforçador da contingência operante. No presente estudo, as seguintes perguntas foram feitas: (1) Uma condição de estímulo antecedente análoga ao SD assumiria função evocativa sobre CCEs e seu produto agregado em uma metacontingência? (2) Quais seriam os efeitos de alternar duas condições de estímulo antecedentes, sendo cada uma delas correlacionada a uma metacontingência específica? (3) O estabelecimento do controle de estímulos análogo ao discriminativo produziria processos análogos ao de generalização? Participaram do estudo 15 estudantes universitários, totalizando 12 gerações (3 participantes em cada). O estudo teve sete condições experimentais em procedimento de tentativas. Um a três participantes trabalhavam simultaneamente, cada um em um computador: o participante da esquerda (PE) do centro (PC) e da direita (PD). Em cada tentativa, nas telas dos computadores de cada participante, eram apresentados, independentemente, quatro números um em cada coluna e cabia ao participante inserir outros quatro também em quatro colunas. Se as somas dos números apresentados pelos computadores e dos números selecionados em cada coluna pelo participante resultassem em números ímpares, o participante ganhava pontos. Esta contingência foi chamada de contingência individual. Outras duas contingências incidiam sobre relações entre os produtos dos comportamentos dos participantes, chamadas de metacontingências. Na metacontingência 1, a cor de fundo da tela era azul (SM1) e se a soma dos 4 números inseridos pelo participante PE fosse menor que a soma dos 4 números inseridos pelo participante PC e esta soma fosse menor que a soma dos 4 números inseridos por PD ( ∑PE ∑PC ∑PD ), uma conseqüência cultural era produzida: os participantes recebiam créditos adicionais chamado bônus. Na metacontingência 2, as telas tinham fundo vermelho (SM2) e os participantes produziam bônus se ∑PE ∑PC ∑PD . Estas metacontingências eram apresentadas em ordem aleatória entre tentativas. Atingidos critérios de estabilidade, os participantes antigos eram substituídos por participantes novatos. Em momentos pré-estabelecidos foram conduzidos testes de controle de estímulos e de generalização. Os resultados mostraram a seleção do comportamento operante e das/ pelas metacontingências 1 e 2 e indicaram que SM1 e SM2 adquiriram função evocativa sobre as metacontingências correspondentes. Os testes sugeriram que as cores de fundo da tela tornaram-se a dimensão do estímulo que exerceu controle sobre os comportamentos dos participantes e suas interações. Os dados são discutidos em termos de uma analogia entre contingência operante e metacontingência

Cancer is one of the leading causes of death in the world, often due to failed treatments because of drug resistance. Treatment is difficult as resistance is hard to detect before treatment and can develop during treatment. The fluorometric microculture cytotoxicity assay (FMCA) is a reliable, rapid method for testing drug cytotoxicity but requires large cell samples, which can be challenging to obtain. Patient-derived cancer cells (PDC) have proven challenging to culture in monolayer models, but recent studies have shown the possibility of using tumoroids. Tumoroids are three-dimensional models where cells are grown in basement membrane matrix hydrogel, allowing scaffold growth like in vivo tumors. This study aimed to culture colorectal PDC in the form of tumoroids, transfecting them, and examine cell cycle and tumor resistance for 5-Fluorouracil, Oxaliplatin and Irinotecan. Cells were deposited in gels with medium mimicking in vivo conditions, supporting growth and allowing extracellular signaling. The study succeeded in culturing both untransfected and transfected cells, resulting in cells expanding 48 and 42 times, respectively. Cell cycle remained unchanged. No changes were observed in 5-Fluorouracil, but a change was seen in transfected cells at passage 3 with oxaliplatin. The cells showed a 22% difference in survival indexes compared to naïve cells. Changes were seen in Irinotecan’s half maximal inhibitory concentration (IC50); all cell passage IC50 values differed &gt;15.17 µM (p-value 0.0184). In conclusion, PDC can be cultured as tumoroids, but more studies are needed to determine if the model can generate reliable results representing PDC regarding tumor resistance.

Modern techniques like automated microscopy and spectroscopy now make it possible to study quantitatively, across multiple phenotypic and molecular parameters, how cell populations are affected by different treatments and/or environmental disturbances. As the technology development at the instrument level often is ahead of the data analytical tools and the scientific questions, there is a large and growing need for computational algorithms enabling desired data analysis. These algorithms must have capacity to extract and process quantitative dynamic information about how the cell population is affected by different stimuli with the final goal to transform this information into development of new powerful therapeutic strategies. In particular, there is a great need for automated systems that can facilitate the analysis of massive data streams for label-free methods such as phase contrast microscopy (PCM) imaging and spectroscopy (NMR). Therefore, in this thesis, algorithms for quantitative high-throughput phenotypic and systemic profiling of in vitro growing cell populations via label-free microscopy and spectroscopy are developed and evaluated. First a two-dimensional filter approach for high-throughput screening for drugs inducing autophagy and apoptosis from phase contrast time-lapse microscopy images is studied. Then new methods and applications are presented for label-free extraction and comparison of time-evolving morphological features in phase-contrast time-lapse microscopy images recorded from in vitro growing cell populations. Finally, the use of dynamic morphology and NMR/MS spectra for implementation of a reference database of drug induced changes, analogous to the outstanding mRNA gene expression based Connectivity Map database, is explored. In conclusion, relatively simple computational methods are useful for extraction of very valuable biological and pharmacological information from time-lapse microscopy images and NMR spectroscopy data offering great potential for biomedical applications in general and cancer pharmacology in particular.

Background: Cancer is a common disease, and the choice of treatment becomes more difficult over time due to chemotherapy resistant in cancer cells. To improve the in vitroassay and the individual cancer treatment, a luminescence-based endpoint assay, CellTiter Glo 2.0 was compared with the currently in use fluorescence endpoint assay, fluorometric microculture cytotoxic assay.  Aim: The aim of this study was to validate and compare the CellTiter Glo 2.0 assay with awell-established method (FMCA) and MTT [3-(4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2Htetrazolium bromide] assay. Moreover, investigate whether the generated data can be used as a reference database for validation of patient samples in the future.   Materials and methods: The validation was performed on peripheral blood mononuclear cells from different healthy donors and two cell lines (HCT116-wt and HT-29) of colorectal cancer carcinoma were ordered frozen from American Type Culture Collection. Analysis was also done in solid samples (ovarian and kidney cancer cells). To get as correct evaluation as possible all materials were analyzed in parallel between the two methods.  Results and conclusion: A clear trend was observed when using CellTiter Glo 2.0 assay,post FMCA directly on tumor cells. This setup, makes it possible to collect reference data in the future. In addition, a high spread of the survival index data was noted between the two methods. The reason is still unknown but could be due to the low number of tested tumor cells, therefore more tumor cells need to be tested in future studies