Designing in the Wild: Towards Specialized Clothing Designs

Designing clothing is often designer-centric, especially when creating fashion wear, ready-to-wear clothing and accessories. Fashion designers are expected to peruse the market, speculate on new styles and forecast trends with a vision to realizing designs that will sell. Designs for clothing often seem to come from ‘within’ the designer and are a reflection of what has been previously created and what might be considered desirable in the future. Alternatively, with the growing needs for more specialized designs due to aging demographics, specialized work situations and people with special needs it is almost impossible to predict the trends and market without understanding authentic human behavior

Nuclear architecture explored by live-cell fluorescence microscopy using laser and ion microbeam irradiation.

Nuclear architecture is a biological field of research that studies the spatio-temporal organization of the components within cell nuclei. Since nuclei are the organelles that harbor the genome and epigenome, they are the place where most of the genetic processes like replication, transcription, splicing, gene-regulation, DNA repair, re- combination etc. are carried out. In the presented doctoral thesis modern 4D live-cell microscopy in combination with laser or ion microbeam irradiation (to label or damage chromatin, respectively) was used to study nuclear architecture in living cells over extended periods of time at the single cell level. The results presented in this thesis can be partitioned into three main parts: (a) chromatin dynamics in cycling cells, (b) adaptation of the ion micro beam facil- ity SNAKE to the needs of live-cell observation (including first experiments) and (c) exploring spatio-temporal dynamics of DNA repair proteins after laser micro ir- radiation. (A) Chromatin dynamics in cycling cells Distribution of interphase chromosomes within cell nuclei has been found to be non- random with respect to gene density and chromosome size. Changes in nuclear orga- nization have been reported in several disorders and diseases. To which extent relative chromosome positioning is conserved through mitosis in cycling cells and whether certain chromatin domains are able change their relative position dramatically in the interphase nucleus has been the subject of various mechanistic models and contro- versial discussions. In 1909 German biologist theodor Boveri was the first one to comment on this topic in his publication: “Die Blastomerenkerne von Ascaris mega- locephala und die Theorie der Chromosomenindividualität” (included as an appendix to this thesis). In order to test Boveri’s hypotheses, 4D live-cell observations were carried out on a modern spinning disc confocal microscope using a human cell line that possesses photoactivatable chromatin. In experiments that used photoactivation and photobleaching of chromatin, it could be demonstrated that – as stated by Boveri – chromatin proximity relationships are in general not conserved through mitosis but destroyed during early prometaphase by the mechanics of mitosis. Other experiments showed that nuclear rotations in a conveyer-belt-like manner are able to bring initially distant chromatin domains into close proximity in a matter of a few minutes. (B) Adaptation of the SNAKE micro beam facility to the needs of live- cell microscopy (including first experiments) Since ordinary irradiation sources lack the ability to perform targeted micro irradia- tion at the micrometer scale and laser micro irradiation produces an artificial mix of various DNA damages, the ion microbeam SNAKE represents an interesting tool to explore the dynamics of repair proteins in a spatio-temporal context. In the course of a collaboration project the ion microbeam was adapted to the needs of long-term live-cell microscopy. These adaptations and first live-cell experiments performed at the refurbished ion micro beam are described in this part of the results. (C) Exploring spatio-temporal dynamics of DNA repair proteins after laser micro irradiation. Mutation of genetic information can cause serious harm to a cell or even a whole or- ganism. DNA repair serves to protect and clean the genome from undirected poten- tially hazardous changes. Compared to the wealth of information which is available about DNA repair at the molecular level only little attention has been payed to it in context of nuclear architecture. In the last part of the results cells stably expressing GFP tagged versions of the repair proteins MDC1, Rad52 and 53BP1 were damaged by laser micro irradiation and imaged over extended periods of time. It could be de- monstrated that at the used damage induction conditions most of the cells show only minor changes with respect to localization of damage signals, kinetochores and nu- cleoli pattern over time. Furthermore, disappearance of spontaneous 53BP1-GFP foci in favor of protein recruitment to damaged chromatin and mutual exclusion between kinetochore signals and Rad52-GFP damage foci could be observed. In a few U2OS Rad52-GFP nuclei DNA damage foci disappeared simultaneously after a dramatic phase in which the total number of foci drastically increased – even adjacent to the laser damaged chromatin

Regulation of Cell Polarization and Map Kinase Signaling in the Saccharomyces Cerevisiae Pheromone Response Pathway: a Dissertation

Exposure to external stimuli promotes a variety of cellular responses including changes in morphology, gene expression and cell division status. These responses are promoted by signaling pathways composed of modules that are conserved from lower to higher eukaryotes. In Saccharomyces cerevisiae response to the external stimuli provided by mating pheromone is governed by the pheromone response pathway. This pathway is composed of a G protein coupled receptor/heterotrimeric G protein (Gαβγ) module and a MAP kinase cascade. Activation of this pathway allows the heterotrimeric G protein βγ dimer (Gβγ) to recruit polarity proteins to promote changes in cell morphology and to activate signaling through the MAP kinase cascade. Here we investigate the regulation of these pheromone-induced responses. We first examine how an asymmetric polarization response is generated. Normally, a gradient of pheromone serves as a spatial cue for formation of a polarized mating projection, but cells can still polarize when pheromone is present uniformly. Here we show that an intact receptor/Gαβγ module is required for polarization in response to both a gradient and uniform concentration of pheromone. Further investigation into regulation of Gβγ by Gα revealed that the two interaction interfaces between Gα and Gβ have qualitatively different roles. Our results suggest that one interface controls signaling whereas the other governs coupling to the receptor. Overall our results indicate that communication between the receptor and Gαβγ is required for proper polarization. We then examine how G1 CDKs regulate MAP kinase signaling. Response to pheromone is restricted to the G1 stage of the cell cycle. Once cells commit to a round of division they become refractory to mating pheromone until that round of division is complete. One contributor to this specificity involves inhibition of signaling through the MAP kinase cascade by G1 CDKs, but it was not known how this occurs. Here, we show that the MAP kinase cascade scaffold Ste5 is the target of this inhibition. Cln/CDKs inhibit signaling by phosphorylating sites surrounding a small membrane-binding domain in Ste5, thereby disrupting the membrane localization of Ste5. Furthermore, we found that disrupting this regulation allows cells to arrest at an aberrant non-G1 position. Our findings define a mechanism and a physiological benefit for restricting pheromone-induced signaling to G1. This thesis describes findings related to generation of an asymmetric polarization response, heterotrimeric G protein function, and coordination of differentiation signaling with cell division status. Lessons learned here might be applicable to the regulation of polarization and differentiation responses in other systems as the signaling modules are conserved

Quantifying False Positives in Avian Survey Data

Imperfect detection is a known issue when conducting count-based surveys in wildlife studies. False positive detections, observed occurrences of individuals that truly are not present, are often assumed to not occur. This assumption can bias detection rates and create misleading results when calculating population estimates. Survey methods such as the dependent double-observer method are suggested to reduce the occurrence of false positives (Nichols et al. 2000). My study quantified and compared rates of false positives in a single-observer method and a dependent double-observer method using computer-generated auditory surveys. I categorized volunteer observers as either inexperienced or experienced and asked them to identify vocalizations of ten grassland songbird species native to central Montana. False positive rates of experienced observers declined from 0.095 in single-observer surveys to 0.032 in dependent double-observer surveys. False positive rates of inexperienced observers declined from 0.511 in single-observer surveys to 0.391 in dependent double-observer surveys. Further evaluation will provide information on the effectiveness of the dependent double-observer method in providing more precise and less biased population estimates

Quantifying False Positives in Avian Survey Data

Imperfect detection is a known issue with conducting wildlife surveys. False positive detections, where an individual is counted as present when it truly is not, are often assumed to not occur. This assumption can skew detection rates and create misleading results when calculating population estimates. Survey methods such as the dependent double-observer method developed by Nichols et al. (2000) are suggested to reduce the occurrence of false positives by using two collaborating observers. This study quantified and compared rates of false positives between a single-observer method and the dependent double-observer method. This was accomplished with auditory surveys of ten grassland songbird species native to central Montana. Both inexperienced and experienced volunteer observers were asked to listen to randomly-generated surveys containing the vocalizations of these ten songbirds and identify the species. The decrease in false positive rates using the dependent double-observer method is substantial. Further evaluation will provide information on the effectiveness of the dependent double-observer method in providing more precise and less biased population estimates

Advances in Workwear: Protection from Steam & Hot Water

The design objective was to create a garment system using semi-permeable fabric to protect workers from steam and hot water injuries

For the Love of Sewing

The purpose of this study was to explore why professional women who are expert sewers love to sew, while at the same time determine how they perceived and identified with their acts of making. We were interested in understanding why women love working with fabrics, textiles, threads and needles and immersing themselves in making things

Do You See What I See? Using Ethnographic Methods to Inform Functional Design

All people wear clothes, but dressing is an activity taken for granted until balance and synchronized movements required to do so are lost due to illness, injury, disease, or surgery. Ethnographic methods were used to map the use scenario, examine the clothing context and its meaning to people through field observation of therapy sessions and patient routines with personal support workers (n=46), and interviews with therapists, care workers, and patients (n=34). Results reveal patients\u27 experience related to clothing, disability, and functioning as well as the psychological aspects of clothing. Findings include design recommendations to mediate difficulties people have when dressing through consideration of fabric choices, garment silhouettes, circumference of garment openings, garment fasteners, dual waistbands, pockets, loops, and visual clues to guide garment orientation and product development opportunities. Results of the study may impact fashion designers, specialized product developers, design educators, and rehabilitation therapists