Im Rahmen dieser Arbeit wurden Fibroblasten und Keratinozyten, welche in vitro auf unterschiedlichen Scaffolds sowohl gemeinsam als auch in Monokulturen gezüchtet wurden, mittels Real-time PCR auf ihre Genausschüttung untersucht, um festzustellen wie sich die Unterlage auf die Genausschüttung auswirkt. Hierzu wurden die Proben sowohl auf die Genexpressionsmarker für die Basallamina Kollagen IV, Laminin 1 und 5 als auch auf die Genexpressionsmarker für die frühe Differenzierung Keratin K13 und K14 untersucht. Als Referenzgen wurde β-Actin ausgewählt, da dieses Gen in den Vorversuchen mit zwei weiteren Referenzgenen die stabilste Expression gezeigt hatte. Die Genexpressionsanalyse zeigte, dass nur in den Kokulturen von Keratinozyten und Fibroblasten eine ausgewogene Genexpression stattfindet, da sich die Zellen darin beeinflussen und regulieren. / Fibroblasts and keratinocytes were cultured in vitro on different scaffolds in monocultures and cocultures and examined by RT-qPCR for gene expression. Gene expression analysis was made for genes coding for basement Membrane collagen IV, laminin 1 and 5 and for early differentiation keratin K13 and K14. β-Actin was used as reference gene, because it showed in preliminary tests with two other reference genes most stable expression. Gene expression analysis showed only in cocultures of fibroblasts and keratinocytes balanced gene expression, because the two cell types affect and regulate each other.

No description available.

No description available.

The difference in evolutionary interests of males and females can select for traits that favour an individual??s fitness at the expense of their mate??s lifetime fitness. Despite the widespread occurrence of this sexual conflict over reproductive interactions, however, research to date has largely focused on the fitness costs imposed on females by manipulative males. Empirical evidence is particularly sparse for how mating can also be costly for males, the genetic structure of traits involved in reproductive interactions, and how sexual conflict can modify sexual selection in general. My aim was to explore the broader evolutionary consequence of sexual conflict and male-female interactions. In the nuptial-feeding Australian ground cricket, Pteronemobius sp., I used an experimental evolution approach to explore how diet and sexual conflict interact to determine the costs of mating. In the Australian black field cricket, Teleogryllus commodus, I used molecular and quantitative genetic approaches to characterise the fitness consequences and genetic basis of spermatophore attachment, a trait at the centre of inter-locus sexual conflict, and then related this to both condition and male attractiveness. Finally, in T. commodus, I quantified how sexual conflict alters the sexual selection acting on male sexual traits and how this in turn shapes genetic architecture and the persistence of additive genetic variance. My results demonstrate the complex nature of reproductive interactions between males and females. Importantly, I show that diet can mediate the expression of sexual conflict in a mating system and shape the evolution of male life-span. I also show that reproductive interactions influence the fitness benefits that both male and females obtain from mating in ways that are not predicted by current theory and that much of the potential for such traits to co-evolve is via a common genetic association with condition. Finally, I demonstrate that sexual conflict can profoundly modify the process and outcome of sexual selection, thereby influencing how additive genetic variation is maintained in a suite of male sexual traits. These results highlight the need for a greater integration of sexual conflict and sexual selection theory as the evolutionary potential and significance of sexual conflict may currently be underestimated.

Doctor of Philosophy (PhD) / GABAA receptors are implicated in the pathology of psychiatric disorders such as schizophrenia and depression. They are rapidly affected by stress in a sex-dependent fashion, suggesting that GABAA receptors may be relevant to understanding the association between stress and psychiatric disorders. Thus, this thesis examined how GABAA receptors are affected in both male and female mice exposed to stress in adulthood (Chapter 2), early-life (Chapter 3-5) and a combination of both early-life and adulthood stress (Chapter 6). 2. The effects of acute adulthood stress (3 minute warm swim stress) on GABAA receptor binding in the brains of male and female mice were examined using quantitative receptor autoradiography. The total number of GABAA receptor [3H]GABA binding sites was increased following swim stress in specific forebrain cortical regions of female mice swum individually or in a group, but decreased in male mice when swum in a group only. These findings confirm and extend previous studies, identifying the cortical regions involved in rapid stress-induced changes in GABAA receptors. 3. Post-natal handling models in rodents comparing control (brief handling sessions; EH) with no intervention stress conditions (NH), indicate that the NH condition results in an anxious adulthood phenotype and this was confirmed in the present thesis using the elevated plus-maze behavioural test. Using this model the effects of early-life stress on adulthood GABAA receptors were then examined. 4. Regional densities of GABAA receptor α1 and α2 subunit proteins were observed in the adult brain of male and female mice using immunoperoxidase histochemistry. NH males showed a loss of the α2 subunit from the thalamus and the lower layers (IV-VI) of the primary somatosensory cortex, whilst NH females showed a reduction of α2 but an increase in α1 protein in the lower layers of the primary somatosensory cortex only. These regionally specific alterations in the α1:α2 subunit ratio suggest that early-life stress disrupts the developmental α subunit switch, which occurs in a regionally-dependent fashion over the first two weeks of rodent life. 5. Double-labelling immunofluorescence and confocal microscopy were used to examine the effects of sex and early-life stress on GABAA receptor synaptic clustering. Regardless of sex, mice exposed to early-life stress (NH) showed reduced colocalisation of the GABAA receptor α2 subunit with the synaptic marker protein gephyrin relative to the control condition (EH). This suggests that early-life stress impairs adulthood inhibitory synaptic strength and is consistent with the increased anxiety of the stressed relative to control mice. 6. Finally, the effects of early-life stress on adulthood swim stress-induced changes in GABAA receptor binding were examined using quantitative receptor autoradiography in forebrain cortical regions. Findings showed that the effect of adulthood stress on the total number of GABAA receptor binding sites for [3H]GABA in forebrain cortical regions was altered by early-life stress in both male and female mice, suggesting that the rapid adulthood stress response of GABAA receptors is affected by early-life experience. 7. Together these results show that GABAA receptors are sensitive to subtle changes in the environment in both early-life and adulthood and that these neurochemical responses to stress in adulthood are sex-dependent. The short and long-term stress-sensitivity of the GABAergic system implicates GABAA receptors in the non-genetic aetiology of psychiatric illnesses in which sex and stress are important factors.

Ascochyta blight, caused by Ascochyta rabiei (Pass.) Labrousse, is one of the most destructive diseases of chickpea (Cicer arietinum L.) worldwide. Despite the existence of highly resistant uncultivated genotypes, attempts to develop cultivars with a high level of durable resistance have been unsuccessful. This study investigated the chickpea defence response to A. rabiei using a functional genomics approach, which has the capacity to improve the overall understanding of the coordinated defence response at a molecular level. An existing cDNA library was used to generate a resource of Expressed Sequence Tags (ESTs) that, after clustering, comprised 516 unigenes. The unigenes were functionally annotated resulting in the identification of 20 specific defence-related unigenes, as well as numerous transcripts with possible involvement in the coordination of defence responses. To explore the expression patterns of the defence-related unigenes in an A. rabiei resistant and susceptible genotype, the unigenes were employed as probes in microarrays. Resulting expression data was analysed to identify differentially expressed unigenes over a time-course after infection. Comparison of the expression profiles from the resistant and susceptible genotype identified three putative genes that were exclusively up-regulated in the resistant genotype, thus may be involved in an effective defence response. Considering that a defence response can involve hundreds of genes, the entire set of chickpea unigenes were used to construct large-scale microarrays. To supplement the chickpea probes, 156 putative defence-related grasspea (Lathyrus sativus L.) ESTs and 41 lentil (Lens culinaris Med.) Resistance Gene Analogs (RGAs) were also included. Expression profiles for three chickpeas and one wild relative were generated over a time course. 97 differentially expressed ESTs were identified using a robust experimental system that included confirmation by quantitative RT-PCR. The results indicated that genes involved in the active defence response were similar to those governed by R-gene mediated resistance, including the production of reactive oxygen species and the hypersensitive response, down-regulation of 'housekeeping' gene expression, and expression of pathogenesis-related proteins. The comparison between resistant and susceptible genotypes identified certain gene expression 'signatures' that may be predictiv e of resistance. To further characterise the regulation of potential defence-related genes, the microarray was used to study expression profiles of the three chickpea genotypes (excluding the wild relative) after treatment with the defence signalling compounds, ethylene (E), salicylic acid (SA), and jasmonate (JA). 425 ESTs were differentially expressed, and comparison between genotypes revealed the presence of a wider range of inducible defence responses in resistant genotypes. Linking the results with the previous microarray results indicated the presence of other pathogen-specific signalling mechanisms in addition to E, SA and JA. The lower arsenal of defence-related gene expression observed in the susceptible genotype may be a result of 'breaks' in the pathways of defence-related gene activation. To draw together the findings of all experiments, a model was constructed for a hypothetical mechanism of chickpea resistance to A. rabiei. The model was synthesised based on the evidence gathered in this study and previously documented defence mechanisms in chickpea, and identified signal transduction as a key to resistance.

The lifting associated with childcare has been linked to musculoskeletal disorders (MSDs) in mothers (Griffin & Price, 2000; Sanders & Morse, 2005). The purpose of the current study was to investigate, for the first time through observation, the lifting of young children by mothers in the home environment to identify risk factors which might give rise to MSDs. Twenty five mothers with one or two children weighing between 9 and 15kg (n = 30) completed a self-report survey modified for the New Zealand context (The Ergonomics of Caring for Children (Sanders & Morse, 2005)). They also took part in a structured observation of lifting in the home using a checklist based on the New Zealand Manual Handling Hazard Control Record NZMHHCR (OSH & ACC, 2001). Modifications were derived from careful consultation of current literature to enable contributory factors related to the load, the mother, the environment and the task to be assessed and a rating of low, medium, or high risk to be assigned to each factor. A protocol to guide risk assessment was developed to accompany the tool. The modified observational checklist was named the OMLITH (Observing Mothers Lifting In The Home). Survey data identified the mothers as aged between 28 and 40 years, predominantly NZ European, and of average height (χ = 1.69cm) and weight (Body Mass Index = 24). The children in the sample weighed between 9 and 14.5kg and had an average age of 17months. All the mothers were either married (n=20) or living with a significant other (n=5) and their partners frequently (n=13) or always (n=12) helped with childcare. The mean time mothers spent per week on the following activities were: sleep, 6.8hrs (n=25), housework, 14.9hrs (n=25), exercise, 2.9hrs (n=22), watching television or using the home computer, 4.3hrs (n=22), hobbies, 2.2hrs (n=19), gardening or home maintenance, 2.9hrs (n=18). Thirteen of the mothers worked (3 full-time, 10 part-time) and 20 mothers used childcare services. Ratings of the physical stress associated with 50 childcare tasks showed that mothers differentiated between tasks in terms of physical stress to a significant degree (p.000), typically rating ‘Bending while carrying a child’ as almost twice as stressful as the average rating. Other categories rated significantly above the average stress rating were: ‘Use of a backpack to carry infant/child’, ‘Use of baby jogger’, ‘Carrying child on your shoulders’, ‘Standing bent over to wash child in bath or sink’, ‘Lifting child into or out of cot’, ‘Prolonged squatting or stooping while playing with child’, and ‘Placing child in car seat or removing child from car seat’. Mothers reported experiencing MSDs most commonly in the low back (n=16), and the neck, shoulder and upper back (n=8 each). Data from the 87 observations using the OMLITH showed that children were a challenging load due to their moving centre of gravity, and were often unpredictable or awkward to handle. The grip required to lift a child more often than not fell outside the National Institute for Occupational Safety and Health (NIOSH) recommendation. In 72.9% of tasks the child’s weight created a high risk level. The lifting tasks involved horizontal and vertical lift distances that presented a moderate to high risk level in 82.8% of situations. Lifting while twisting and side-bending was assigned a moderate to high risk level in 72.4% of tasks. Risk associated with working at an externally controlled pace; and handling children while seated or kneeling/crouching was also observed. The home environment presented risk associated with obstacles, a variety of floor surfaces and stairs or slopes. Mothers were also observed lifting in confined spaces. Individual factors identified as important considerations were: a mismatch between mothers’ strength and fitness and the lifting requirements, pain or injury, pregnancy, and fatigue. The structured checklist proved to be an appropriate tool to identify the contributory risk factors present when mothers lift in the home and to make an assessment of the level of risk. Results suggest a notable number of risk factors which might give rise to MSDs are present when mothers lift their children at home. The author concludes that further research is warranted to quantify risks, to identify prevention strategies for MSDs in this population, and to guide health providers with regard to treatment and rehabilitation of mothers with MSDs.

Title from container. Includes bibliographies and indexes. Contributions to quantitative and population genetics -- The biochemical genetics of man -- The theory of plant breeding -- Natural selection and its constraints.

This thesis concerns the identification of quantitative trait loci (QTL) for important traits in cattle line crosses. One of these traits is birth weight of calves, which affects both animal production and welfare through correlated effects on parturition and subsequent growth. Birth weight was one of the traits measured in the Davies' Gene Mapping Project. These data form the motivation for the methods presented in this thesis. Multiple QTL models have been previously proposed and are likely to be superior to single QTL models. The multiple QTL models can be loosely divided into two categories : 1 ) model building methods that aim to generate good models that contain only a subset of all the potential QTL ; and 2 ) methods that consider all the observed marker explanatory variables. The first set of methods can be misleading if an incorrect model is chosen. The second set of methods does not have this limitation. However, a full fixed effect analysis is generally not possible as the number of marker explanatory variables is typically large with respect to the number of observations. This can be overcome by using constrained estimation methods or by making the marker effects random. One method of constrained estimation is the least absolute selection and shrinkage operator (LASSO). This method has the appealing ability to produce predictions of effects that are identically zero. The LASSO can also be specified as a random model where the effects follow a double exponential distribution. In this thesis, the LASSO is investigated from a random effects model perspective. Two methods to approximate the marginal likelihood are presented. The first uses the standard form for the double exponential distribution and requires adjustment of the score equations for unbiased estimation. The second is based on an alternative probability model for the double exponential distribution. It was developed late in the candidature and gives similar dispersion parameter estimates to the first approximation, but does so in a more direct manner. The alternative LASSO model suggests some novel types of predictors. Methods for a number of different types of predictors are specified and are compared for statistical efficiency. Initially, inference for the LASSO effects is performed using simulation. Essentially, this treats the random effects as fixed effects and tests the null hypothesis that the effect is zero. In simulation studies, it is shown to be a useful method to identify important effects. However, the effects are random, so such a test is not strictly appropriate. After the specification of the alternative LASSO model, a method for making probability statements about the random effects being above or below zero is developed. This method is based on the predictive distribution of the random effects (posterior in Bayesian terminology). The random LASSO model is not sufficiently flexible to model most QTL mapping data. Typically, these data arise from large experiments and require models containing terms for experimental design. For example, the Davies' Gene Mapping experiment requires fixed effects for different sires, a covariate for birthdate within season and random normal effects for management group. To accommodate these sources of variation a mixed model is employed. The marker effects are included into this model as random LASSO effects. Estimation of the dispersion parameters is based on an approximate restricted likelihood (an extension of the first method of estimation for the simple random effects model). Prediction of the random effects is performed using a generalisation of Henderson's mixed model equations. The performance of the LASSO linear mixed model for QTL identification is assessed via simulation. It performs well against other commonly used methods but it may lack power for lowly heritable traits in small experiments. However, the rate of false positives in such situations is much lower. Also, the LASSO method is more precise in locating the correct marker rather than a marker in its vicinity. Analysis of the Davies' Gene Mapping Data using the methods described in this thesis identified five non-zero marker-within-sire effects ( there were 570 such effects). This analysis clearly shows that most of the genome does not affect the trait of interest. The simulation results and the analysis of the Davies' Gene Mapping Project Data show that the LASSO linear mixed model is a competitive method for QTL identification. It provides a flexible method to model the genetic and experimental effects simultaneously. / Thesis (Ph.D.)--School of Agriculture, Food and Wine, 2006.

L'objectif de la présente thèse est de mettre au jour la richesse de la pensée monétaire d'Edwin Walter Kemmerer (1875-1945). Nous soulignons sa contribution à la théorie quantitative de la monnaie, à la théorie de la banque centrale et à l'analyse de l'étalon-or. Nous présentons “le médecin” et théoricien de la monnaie en quatre chapitres. Le premier aborde sa formation intellectuelle et les idées politiques qui accompagnaient ses propositions de réforme monétaire et bancaire. Le deuxième chapitre analyse l'équation d'échange, la notion de Business confidence et le « test » statistique construit pour valider la théorie quantitative. Le troisième chapitre traite de la participation de Kemmerer au processus de création de la Réserve fédérale américaine en 1913 : sa vision des problèmes persistant sous le National Banking System de 1863, son étude du marché monétaire américain en 1910, son plaidoyer, et ses propositions, pour la future banque centrale. Enfin, le quatrième chapitre expose les différents types d'étalon-or proposés par Kemmerer, mis en oeuvre dans certaines colonies et dans certains pays de l'Amérique latine, en particulier le Gold Exchange Standard dont il est un des précurseurs. La conclusion aborde la question de l'oubli dont Kemmerer est l'objet.