Preimplantation genetic diagnosis: Analysis of gene expression, nuclear DNA and cytoplasmic DNA

Preimplantation genetic diagnosis (PGD) following in vitro fertilization (IVF) offers couples at risk for transmitting genetic disorders the opportunity to identify affected embryos prior to replacement. In particular, embryo gender determination permits screening for X-linked diseases of unknown etiology. Analysis of embryos can be performed by polymerase chain reaction (PCR) amplification of material obtained by micromanipulation. This approach provides an alternative to the termination of an established pregnancy following chorionic villi sampling or amniocentesis. ^ Lately, the focus of preimplantation diagnosis and intervention has been shifting toward an attempt to correct cytoplasmic deficiencies. Accordingly, it is the aim of this investigation to develop methods to permit the examination of single cells or components thereof for clinical evaluation. In an attempt to lay the groundwork for precise therapeutic intervention for age related aneuploidy, transcripts encoding proteins believed to be involved in the proper segregation of chromosomes during human oocyte maturation were examined and quantified. Following fluorescent rapid cycle RT-PCR analysis it was determined that the concentration of cell cycle checkpoint gene transcripts decreases significantly as maternal age increases. Given the well established link between increasing maternal age and the incidence of aneuploidy, these results suggest that the degradation of these messages in aging oocytes may be involved with inappropriate chromosome separation during meiosis. ^ In order to investigate the cause of embryonic rescue observed following clinical cytoplasmic transfer procedures and with the objective of developing a diagnostic tool, mtDNA concentrations in polar bodies and subcellular components were evaluated. First, the typical concentration of mtDNA in human and mouse oocytes was determined by fluorescent rapid cycle PCR. Some disparity was noted between the copy numbers of individual cytoplasmic samples which may limit the use of the current methodology for the clinical assessment of the corresponding oocyte. ^

Development of ``HealthServe'': A measure of customer service orientation in healthcare

An assessment tool designed to measure a customer service orientation among RN's and LPN's was developed using a content-oriented approach. Critical incidents were first developed by asking two samples of healthcare managers (n = 52 and 25) to identify various customer-contact situations. The critical incidents were then used to formulate a 121-item instrument. Patient-contact workers from 3 hospitals (n = 102) completed the instrument along with the NEO-FFI, a measure of the Big Five personality factors. Concurrently, managers completed a performance evaluation scale on the employees participating in the study in order to determine the predictive validity of the instrument.^ Through a criterion-keying approach, the instrument was scaled down to 38 items. The correlation between HealthServe and the supervisory ratings of performance evaluation data supported the instrument's criterion-related validity (r =.66, p $<$.0001). Incremental validity of HealthServe over the Big Five was found with HealthServe accounting for 46% of the variance.^ The NEO-FFI was used to assess the correlation between personality traits and HealthServe. A factor analysis of HealthServe suggested 4 factors which were correlated with the NEO-FFI scores. Results indicated that HealthServe was related to Extraversion, Openness to Experience, Agreeableness, Conscientiousness and negatively related to Neuroticism.^ The benefits of the test construction procedure used here over the use of broad-based measures of personality were discussed as well as the limitations of using a concurrent validation strategy. Recommendations for future studies were provided. ^

Periphyton as an indicator of restoration in the Everglades

Periphyton communities dominate primary production in much of the Florida Everglades wetland and therefore contribute to soil production, ecosystem metabolism and secondary production as well as the composition of dependent communities. Decades of research in the Everglades have supported research findings from other wetland types that cumulatively show that periphyton communities respond very rapidly to alterations in the two dominant drivers of wetland structure and function—hydrology and water quality. Hydrology controls periphyton productivity and composition by regulating moisture availability, substrate types available for colonization and supply of nutrients. Nutrients, particularly the limiting nutrient in this system, phosphorus (P), control levels of production and community composition. Because periphyton communities are well-established to be related to hydrology and water quality, an indicator was developed based on three periphyton attributes: abundance, quality (i.e., nutrient content) and community composition. This assessment tool offers a qualitative assessment of ecosystem response to potential changes in management activities at a time scale appropriate for active management. An example is provided of how the indicator can be used to assess the current water quality and hydrological conditions from high-density spatial surveys. Detected patterns of deterioration align with expectations derived from model predictions and known sources of nutrients and unnatural hydrologic regimes. If employed adaptively in ecosystem management, this tool can be used to both detect and react to change before the system has been irreparably altered.

Hydro-Meteorology and Water Budget of the Mara River Basin Under Land Use Change Scenarios

Mara is a transboundary river located in Kenya and Tanzania and considered to be an important life line to the inhabitants of the Mara-Serengeti ecosystem. It is also a source of water for domestic water supply, irrigation, livestock and wildlife. The alarming increase of water demand as well as the decline in the river flow in recent years has been a major challenge for water resource managers and stakeholders. This has necessitated the knowledge of the available water resources in the basin at different times of the year. Historical rainfall, minimum and maximum stream flows were analyzed. Inter and intra-annual variability of trends in streamflow are discussed. Landsat imagery was utilized in order to analyze the land use land cover in the upper Mara River basin. The semi-distributed hydrological model, Soil and Water Assessment Tool (SWAT) was used to model the basin water balance and understand the hydrologic effect of the recent land use changes from forest-to-agriculture. The results of this study provided the potential hydrological impacts of three land use change scenarios in the upper Mara River basin. It also adds to the existing literature and knowledge base with a view of promoting better land use management practices in the basin.

A Modeling Approach to Determine the Impacts of Land Use and Climate Change Scenarios on the Water Flux of the Upper Mara River

With the flow of the Mara River becoming increasingly erratic especially in the upper reaches, attention has been directed to land use change as the major cause of this problem. The semi-distributed hydrological model Soil and Water Assessment Tool 5 (SWAT) and Landsat imagery were utilized in the upper Mara River Basin in order to 1) map existing field scale land use practices in order to determine their impact 2) determine the impacts of land use change on water flux; and 3) determine the impacts of rainfall (0%, ±10% and ±20%) and air temperature variations (0% and +5%) based on the Intergovernmental Panel on Climate Change projections on the water flux of the 10 upper Mara River. This study found that the different scenarios impacted on the water balance components differently. Land use changes resulted in a slightly more erratic discharge while rainfall and air temperature changes had a more predictable impact on the discharge and water balance components. These findings demonstrate that the model results 15 show the flow was more sensitive to the rainfall changes than land use changes. It was also shown that land use changes can reduce dry season flow which is the most important problem in the basin. The model shows also deforestation in the Mau Forest increased the peak flows which can also lead to high sediment loading in the Mara River. The effect of the land use and climate change scenarios on the sediment and 20 water quality of the river needs a thorough understanding of the sediment transport processes in addition to observed sediment and water quality data for validation of modeling results.

Land Use and Climate Change Impacts on the Hydrology of the Upper Mara River Basin, Kenya: Results of a Modeling Study to Support Better Resource Management

Some of the most valued natural and cultural landscapes on Earth lie in river basins that are poorly gauged and have incomplete historical climate and runoff records. The Mara River Basin of East Africa is such a basin. It hosts the internationally renowned Mara-Serengeti landscape as well as a rich mixture of indigenous cultures. The Mara River is the sole source of surface water to the landscape during the dry season and periods of drought. During recent years, the flow of the Mara River has become increasingly erratic, especially in the upper reaches, and resource managers are hampered by a lack of understanding of the relative influence of different sources of flow alteration. Uncertainties about the impacts of future climate change compound the challenges. We applied the Soil Water Assessment Tool (SWAT) to investigate the response of the headwater hydrology of the Mara River to scenarios of continued land use change and projected climate change. Under the data-scarce conditions of the basin, model performance was improved using satellite-based estimated rainfall data, which may also improve the usefulness of runoff models in other parts of East Africa. The results of the analysis indicate that any further conversion of forests to agriculture and grassland in the basin headwaters is likely to reduce dry season flows and increase peak flows, leading to greater water scarcity at critical times of the year and exacerbating erosion on hillslopes. Most climate change projections for the region call for modest and seasonally variable increases in precipitation (5–10 %) accompanied by increases in temperature (2.5–3.5 °C). Simulated runoff responses to climate change scenarios were non-linear and suggest the basin is highly vulnerable under low (−3 %) and high (+25 %) extremes of projected precipitation changes, but under median projections (+7 %) there is little impact on annual water yields or mean discharge. Modest increases in precipitation are partitioned largely to increased evapotranspiration. Overall, model results support the existing efforts of Mara water resource managers to protect headwater forests and indicate that additional emphasis should be placed on improving land management practices that enhance infiltration and aquifer recharge as part of a wider program of climate change adaptation.

Water Demand and Allocation in the Mara River Basin, Kenya/Tanzania in the Face of Land Use Dynamics and Climate Variability

The Mara River Basin (MRB) is endowed with pristine biodiversity, socio-cultural heritage and natural resources. The purpose of my study is to develop and apply an integrated water resource allocation framework for the MRB based on the hydrological processes, water demand and economic factors. The basin was partitioned into twelve sub-basins and the rainfall runoff processes was modeled using the Soil and Water Assessment Tool (SWAT) after satisfactory Nash-Sutcliff efficiency of 0.68 for calibration and 0.43 for validation at Mara Mines station. The impact and uncertainty of climate change on the hydrology of the MRB was assessed using SWAT and three scenarios of statistically downscaled outputs from twenty Global Circulation Models. Results predicted the wet season getting more wet and the dry season getting drier, with a general increasing trend of annual rainfall through 2050. Three blocks of water demand (environmental, normal and flood) were estimated from consumptive water use by human, wildlife, livestock, tourism, irrigation and industry. Water demand projections suggest human consumption is expected to surpass irrigation as the highest water demand sector by 2030. Monthly volume of water was estimated in three blocks of current minimum reliability, reserve (>95%), normal (80–95%) and flood (40%) for more than 5 months in a year. The assessment of water price and marginal productivity showed that current water use hardly responds to a change in price or productivity of water. Finally, a water allocation model was developed and applied to investigate the optimum monthly allocation among sectors and sub-basins by maximizing the use value and hydrological reliability of water. Model results demonstrated that the status on reserve and normal volumes can be improved to ‘low’ or ‘moderate’ by updating the existing reliability to meet prevailing demand. Flow volumes and rates for four scenarios of reliability were presented. Results showed that the water allocation framework can be used as comprehensive tool in the management of MRB, and possibly be extended similar watersheds.