Self-organizing cultured neural networks: Image analysis techniques for longitudinal tracking and modeling of the underlying network structure

Esta tesis estudia la evolución estructural de conjuntos de neuronas como la capacidad de auto-organización desde conjuntos de neuronas separadas hasta que forman una red (clusterizada) compleja. Esta tesis contribuye con el diseño e implementación de un algoritmo no supervisado de segmentación basado en grafos con un coste computacional muy bajo. Este algoritmo proporciona de forma automática la estructura completa de la red a partir de imágenes de cultivos neuronales tomadas con microscopios de fase con una resolución muy alta. La estructura de la red es representada mediante un objeto matemático (matriz) cuyos nodos representan a las neuronas o grupos de neuronas y los enlaces son las conexiones reconstruidas entre ellos. Este algoritmo extrae también otras medidas morfológicas importantes que caracterizan a las neuronas y a las neuritas. A diferencia de otros algoritmos hasta el momento, que necesitan de fluorescencia y técnicas inmunocitoquímicas, el algoritmo propuesto permite el estudio longitudinal de forma no invasiva posibilitando el estudio durante la formación de un cultivo. Además, esta tesis, estudia de forma sistemática un grupo de variables topológicas que garantizan la posibilidad de cuantificar e investigar la progresión de las características principales durante el proceso de auto-organización del cultivo. Nuestros resultados muestran la existencia de un estado concreto correspondiente a redes con configuracin small-world y la emergencia de propiedades a micro- y meso-escala de la estructura de la red. Finalmente, identificamos los procesos físicos principales que guían las transformaciones morfológicas de los cultivos y proponemos un modelo de crecimiento de red que reproduce el comportamiento cuantitativamente de las observaciones experimentales. ABSTRACT The thesis analyzes the morphological evolution of assemblies of living neurons, as they self-organize from collections of separated cells into elaborated, clustered, networks. In particular, it contributes with the design and implementation of a graph-based unsupervised segmentation algorithm, having an associated very low computational cost. The processing automatically retrieves the whole network structure from large scale phase-contrast images taken at high resolution throughout the entire life of a cultured neuronal network. The network structure is represented by a mathematical object (a matrix) in which nodes are identified neurons or neurons clusters, and links are the reconstructed connections between them. The algorithm is also able to extract any other relevant morphological information characterizing neurons and neurites. More importantly, and at variance with other segmentation methods that require fluorescence imaging from immunocyto- chemistry techniques, our measures are non invasive and entitle us to carry out a fully longitudinal analysis during the maturation of a single culture. In turn, a systematic statistical analysis of a group of topological observables grants us the possibility of quantifying and tracking the progression of the main networks characteristics during the self-organization process of the culture. Our results point to the existence of a particular state corresponding to a small-world network configuration, in which several relevant graphs micro- and meso-scale properties emerge. Finally, we identify the main physical processes taking place during the cultures morphological transformations, and embed them into a simplified growth model that quantitatively reproduces the overall set of experimental observations.

Self-organizing cultured neural networks: Image analysis techniques for longitudinal tracking and modeling of the underlying network structure (versión actualizada)

Esta tesis estudia la evolución estructural de conjuntos de neuronas como la capacidad de auto-organización desde conjuntos de neuronas separadas hasta que forman una red (clusterizada) compleja. Esta tesis contribuye con el diseño e implementación de un algoritmo no supervisado de segmentación basado en grafos con un coste computacional muy bajo. Este algoritmo proporciona de forma automática la estructura completa de la red a partir de imágenes de cultivos neuronales tomadas con microscopios de fase con una resolución muy alta. La estructura de la red es representada mediante un objeto matemático (matriz) cuyos nodos representan a las neuronas o grupos de neuronas y los enlaces son las conexiones reconstruidas entre ellos. Este algoritmo extrae también otras medidas morfológicas importantes que caracterizan a las neuronas y a las neuritas. A diferencia de otros algoritmos hasta el momento, que necesitan de fluorescencia y técnicas inmunocitoquímicas, el algoritmo propuesto permite el estudio longitudinal de forma no invasiva posibilitando el estudio durante la formación de un cultivo. Además, esta tesis, estudia de forma sistemática un grupo de variables topológicas que garantizan la posibilidad de cuantificar e investigar la progresión de las características principales durante el proceso de auto-organización del cultivo. Nuestros resultados muestran la existencia de un estado concreto correspondiente a redes con configuracin small-world y la emergencia de propiedades a micro- y meso-escala de la estructura de la red. Finalmente, identificamos los procesos físicos principales que guían las transformaciones morfológicas de los cultivos y proponemos un modelo de crecimiento de red que reproduce el comportamiento cuantitativamente de las observaciones experimentales. ABSTRACT The thesis analyzes the morphological evolution of assemblies of living neurons, as they self-organize from collections of separated cells into elaborated, clustered, networks. In particular, it contributes with the design and implementation of a graph-based unsupervised segmentation algorithm, having an associated very low computational cost. The processing automatically retrieves the whole network structure from large scale phase-contrast images taken at high resolution throughout the entire life of a cultured neuronal network. The network structure is represented by a mathematical object (a matrix) in which nodes are identified neurons or neurons clusters, and links are the reconstructed connections between them. The algorithm is also able to extract any other relevant morphological information characterizing neurons and neurites. More importantly, and at variance with other segmentation methods that require fluorescence imaging from immunocyto- chemistry techniques, our measures are non invasive and entitle us to carry out a fully longitudinal analysis during the maturation of a single culture. In turn, a systematic statistical analysis of a group of topological observables grants us the possibility of quantifying and tracking the progression of the main networks characteristics during the self-organization process of the culture. Our results point to the existence of a particular state corresponding to a small-world network configuration, in which several relevant graphs micro- and meso-scale properties emerge. Finally, we identify the main physical processes taking place during the cultures morphological transformations, and embed them into a simplified growth model that quantitatively reproduces the overall set of experimental observations.

Controlling cell attachment on contoured surfaces with self-assembled monolayers of alkanethiolates on gold.

This paper describes a method based on experimentally simple techniques--microcontact printing and micromolding in capillaries--to prepare tissue culture substrates in which both the topology and molecular structure of the interface can be controlled. The method combines optically transparent contoured surfaces with self-assembled monolayers (SAMs) of alkanethiolates on gold to control interfacial characteristics; these tailored interfaces, in turn, control the adsorption of proteins and the attachment of cells. The technique uses replica molding in poly(dimethylsiloxane) molds having micrometer-scale relief patterns on their surfaces to form a contoured film of polyurethane supported on a glass slide. Evaporation of a thin (< 12 nm) film of gold on this surface-contoured polyurethane provides an optically transparent substrate, on which SAMs of terminally functionalized alkanethiolates can be formed. In one procedure, a flat poly(dimethylsiloxane) stamp was used to form a SAM of hexadecanethiolate on the raised plateaus of the contoured surface by contact printing hexadecanethiol [HS(CH2)15CH3]; a SAM terminated in tri(ethylene glycol) groups was subsequently formed on the bare gold remaining in the grooves by immersing the substrate in a solution of a second alkanethiol [HS(CH2)11(OCH2CH2)3OH]. Then this patterned substrate was immersed in a solution of fibronectin, the protein adsorbed only on the methyl-terminated plateau regions of the substrate [the tri(ethylene glycol)-terminated regions resisted the adsorption of protein]; bovine capillary endothelial cells attached only on the regions that adsorbed fibronectin. A complementary procedure confined protein adsorption and cell attachment to the grooves in this substrate.

Long-term culture of lymphohematopoietic stem cells.

Pluripotent hematopoietic stem cells (PHSCs) show self-renewal and give rise to all blood cell types. The extremely low number of these cells in primary hematopoietic organs and the lack of culture systems that support proliferation of undifferentiated PHSCs have precluded the study of both the biology of these cells and their clinical application. We describe here cell lines and clones derived from PHSCs that were established from hematopoietic cells from the fetal liver or bone marrow of normal and p53-deficient mice with a combination of four growth factors. Most cell lines were Sca-1+, c-Kit+, PgP-1+, HSA+, and Lin- (B-220-, Joro 75-, 8C5-, F4/80-, CD4-, CD8-, CD3-, IgM-, and TER 119-negative) and expressed three new surface markers: Joro 177, Joro 184, and Joro 96. They did not synthesize RNA transcripts for several genes expressed at early stages of lymphocyte and myeloid/erythroid cell development. The clones were able to generate lymphoid, myeloid, and erythroid hematopoietic cells and to reconstitute the hematopoietic system of irradiated mice for a long time. The availability of lymphohematopoietic stem cell lines should facilitate the analysis of the molecular mechanisms that control self-renewal and differentiation and the development of efficient protocols for somatic gene therapy.

Clinical Implications for Working with Gay Men A Self Psychological Treatment Approach.

Specific training for conducting psychotherapy with gay men is limited for psychologists, particularly when using a Self Psychology theoretical orientation (Robertson, 1996). In fact, psychologists often are faced with conflicting and contradictory points of view that mirror society's condemnation of homosexuality (Robertson, 1996). This paper is written from a self-psychological perspective to address the lack of a constructive body of literature that explains the unique treatment needs which impact gay men. Estimates of the prevalence of male homosexuality have generated considerable debate. A common assumption is that there are homosexual and non-homosexual men. However, scientists have long been aware that sexual responsiveness to others of the same sex, like most human traits, is continuously distributed in the population (Michaels, 1996). Still the presumption exists that such traits are stable within each man over time (Michaels, 1996). Conflating same-sex sexual experiences with a categorization of the man as homosexual is problematic, in that defining sexuality solely on the basis of experience excludes people who fantasize about sex with others of the same sex but never have sexual contact. Thus, most modern conceptions of sexual orientation consider personal identification, sexual behavior, and sexual fantasy (McWhirter, Sanders & Reinisch, 1990). Gay men's mental health can only be understood in the context of homosexuality throughout history, since religious and moral objections to sexual attraction between men have existed for centuries. Men who desired other men were regarded as sinful and depraved if not ill or abnormal, and same sex contacts were not distinguished from lewd behaviors (Weeks, 1989). Although most people, regardless of sexual orientation, have experienced some feelings of personal rejection, rarely do heterosexuals become targets for disapproval based on the nature of their attractions and behaviors relative to the same and to the other sex. For lesbians, bisexuals, and gay men, however, homosexuality becomes the focus of aspects of themselves that make them feel hated and hateful (Isay, 1989). While gay men and lesbians are often considered together because of the same-sex nature of their relationships and the similar issues that they may experience in their treatment within society, there are many issues where they might be best studied separately. Issues involving with health, parenthood, sexuality and perceived roles and status in society, for example, are often related more to gender than to any shared concept of a 'gay and lesbian community'. Many issues surrounding lesbians and lesbian culture will have more to do with women's issues, and some issues involving with gay men will have more to do with the gay male subculture and with masculinity. The author of this paper has limited experience in working with lesbian and bisexual individuals, and although it is likely that some of the concepts articulated in this paper could translate to working with lesbian and bisexual individuals, further research is indicated to examine the beneficence of utilizing a Self Psychological orientation in psychotherapy with lesbian women and bisexual individuals. This paper presents an overview of the literature including historical treatments of homosexuality, the history of Self Psychology, key principles in Self Psychology, research on Self Psychology, identity development models for gay men, and Self Psychological perspectives on identity development related to gay men. The literature review is followed by a section on treatment implications for psychologists seeking to treat gay men, including case vignettes based on work from my own practice. I have preserved the anonymity of clients by changing demographics, and rearranging and combining presenting issues and historical backgrounds among the case examples.

Goal Orientation Profiles and Self-Concept of Secondary School Students

The objective of this study is to identify possible combinations of multiple goals that lead to different goal orientation profiles and to determine whether there are significant group differences in self-concept dimensions. The Achievement Goals Tendencies Questionnaire (AGTQ) and the Self-Description Questionnaire-II (SDQ-II) were administered to a sample of 2,022 students of Compulsory Secondary education, ranging in age from 12 to 16 years (M = 13.81, SD = 1.35). Cluster analysis identified four profiles of motivational goals: a group of students with a generalized high motivation profile, a group of students with generalized low motivation profile, a group of students with a predominance of learning goals and achievement goals, and a last group of students with a predominance of achievement goals and social reinforcement goals. Results reveal statistically significant differences among the profiles obtained regarding self-concept dimensions.

Motivational profiles Spanish students of Compulsory Secondary Education: Differential analysis of academic self-attributions

Academic goals and academic self-attributions are relevant variables in school settings. The objective of this study is to identify whether there are combinations of multiple goals that lead to different motivational profiles and to determine whether there are significant differences between the groups obtained regarding causal attributions of success and failure (ability, effort, or external causes) in Mathematics and Language and Literature, and in overall academic performance. The Goal Achievement Tendencies Questionnaire (AGTQ) and the Sydney Attribution Scale (SAS) were administered to a sample of 2022 students of compulsory secondary education, ranging in age from 12 to 16 years (M = 13.81, SD = 1.35). Cluster analysis identified four motivational profiles: a group of students with a high generalized motivation profile, a group of students with low generalized motivation profile, a group of students with predominance of learning goals and achievement goals, and a final group of students with predominance of social reinforcement goals. Results revealed statistically significant differences between the profiles obtained in academic self-attributions.

Alienation Effects : Performance and Self-Management in Yugoslavia, 1945-89 /

Jakovljevic considers the concept of theatricality as central to understanding the events that took place in Yugoslavia. He examines the country’s trials, state ceremonies and festivals, army maneuvers, propaganda, and pop culture as “rehearsals and temporary enactments of an ideologically formulated future.”.

A discourse on self-limited diseases. Delivered before the Massachusetts medical society, at their annual meeting, May 27, 1835.

Microfilm. Ann Arbor, Mich., University Microfilms [n.d.] (American culture series, Reel 183.12)

Of making one's self beautiful.

Of faces and their making.--Culture without college.--Thorn-bearer.--Recipe for good cheer.--House beautiful.

Gymnastics of the voice; a system of correct breathing in singing and speaking, based upon physiological laws. A self-instructor in the training and use of the singing and speaking voice.

Translation of Gymnastik der Stimme.

The social identity perspective - Intergroup relations, self-conception, and small groups

The historical development, metatheoretical background, and current state of the social identity perspective in social psychology are described. Although originally, an analysis mainly of intergroup relations between large-scale social categories, and more recently an analysis with a strong social cognitive emphasis, this article shows that the social identity perspective is intended to be a general analysis of group membership and group processes. It focuses on the generative relationship between collective self-conception and group phenomena. To demonstrate the relevance of the social identity perspective to small groups, the article describes social identity research in a number of areas: differentiation within groups; leadership; deviance; group decision making; organizations; computer mediated communication; mobilization, collective action, and social loafing; and group culture. These art the areas in which most work has been done and which arc therefore best placed for further developments in the near future.

Loyalty without Conformity: Tailoring Self-perception as a Means of Balancing Belonging and Differentiation

We argue that members of individualist cultures balance their desire to belong with their desire to be different by maintaining a self-image as being loyal but relatively immune to group influence. Consistent with this, in Study 1 there was a strong tendency for people to rate themselves as being more independent (i.e., less conformist) than other people in their college. College students also rated themselves as being highly loyal to the group, however no self - other discrepancies were found on this dimension. This is despite the fact that traits of loyalty were rated more positively than were traits of independence. Study 2 provided evidence that culture influences the pattern of self - other discrepancies. Whereas people from individualist countries self-enhance on independence dimensions, people from collectivist countries self-enhance on loyalty dimensions. Again, these effects could not be explained as being a function of how positive these traits were seen to be, suggesting a cultural explanation rather than a straight forward superiority bias explanation for the observed discrepancies in self - other ratings. Results are discussed in relation to the SCENT model.

Campfire culture and computer culture: Museums of the present

In Australia indigenous peoples have never had a treaty with the dominant cultures; and their on-going marginalisation is some testimony to this. However, they have not languished entirely in a policy free environment: media is one area where some policy advances have been made; but media policy development has experienced a number of problems. It has tended to be monolithic in a situation demanding multi and complex treatments. And funding, as always, never seems sufficient to meet those multi and complex needs. This paper examines a small remote community on the island of Milingimbi off the northern coast of Arnhem Land in Australia's far north. People in East Arnhem Land refer to themselves collectively as Yolngu. This community is not typical of many documented cases of media relations between indigenous and non-indigenous peoples; however, the fact that it tends to overturn much of the conventional scholarship surrounding indigenous peoples and the media, helps shed new light on the inadequacy of not only monolithic media policy, but the inadequacy of media-only approaches to policy. Arguably, the significance of the media in Milingimbi is part of a 'triangulated' relationship between indigenous and dominant cultures. That triangulation also involves appropriate forms of government and education, which coupled with appropriate media appear to offer new ways of seeing self-government alongside relative cultural and economic autonomy.