Biomarkers encompass a wide range of different measurable indicators, representing a tangible link to physiological changes occurring within the body. Accessibility, sensitivity, and specificity are significant factors in biomarker suitability. New biomarkers continue to be discovered, and questions over appropriate selection and assessment of their usefulness remain. If traditional markers of inflammation are not sufficiently robust in their specificity, then perhaps alternative means of detection may provide more information. Epigenetic drift (epigenetic modifications as they occur as a direct function with age), and its ancillary elements, including platelets, secreted microvesicles (MVs), and microRNA (miRNA), may hold enormous predictive potential. The majority of epigenetic drift observed in blood is independent of variations in blood cell composition, addressing concerns affecting traditional blood-based biomarker efficacy. MVs are found in plasma and other biological fluids in healthy individuals. Altered MV/miRNA profiles may also be found in individuals with various diseases. Platelets are also highly reflective of physiological and lifestyle changes, making them extremely sensitive biomarkers of human health. Platelets release increased levels of MVs in response to various stimuli and under a plethora of disease states, which demonstrate a functional effect on other cell types.
A distinct pathovar of Salmonella enterica serovar Typhimurium, ST313, has emerged in sub-Saharan Africa as a major cause of fatal bacteremia in young children and HIV-infected adults. D23580, a multidrug resistant clinical isolate of ST313, was previously shown to have undergone genome reduction in a manner that resembles that of the more human-restricted pathogen, Salmonella enterica serovar Typhi. It has since been shown through tissue distribution studies that D23580 is able to establish an invasive infection in chickens. However, it remains unclear whether ST313 can cause lethal disease in a non-human host following a natural course of infection. Herein we report that D23580 causes lethal and invasive disease in a murine model of infection following peroral challenge. The LD50 of D23580 in female BALB/c mice was 4.7 x 105 CFU. Tissue distribution studies performed 3 and 5 days post-infection confirmed that D23580 was able to more rapidly colonize the spleen, mesenteric lymph nodes and gall bladder in mice when compared to the well-characterized S. Typhimurium strain SL1344. D23580 exhibited enhanced resistance to acid stress relative to SL1344, which may lend towards increased capability to survive passage through the gastrointestinal tract as well as during its intracellular lifecycle. Interestingly, D23580 also displayed higher swimming motility relative to SL1344, S. Typhi strain Ty2, and the ST313 strain A130. Biochemical tests revealed that D23580 shares many similar metabolic features with SL1344, with several notable differences in the Voges-Proskauer and catalase tests, as well alterations in melibiose, and inositol utilization. These results represent the first full duration infection study using an ST313 strain following the entire natural course of disease progression, and serve as a benchmark for ongoing and future studies into the pathogenesis of D23580.
MicroRNAs (miRNAs) regulate gene output by targeting degenerate elements in mRNAs and have undergone drastic expansions in higher metazoan genomes. The evolutionary advantage of maintaining copies of highly similar miRNAs is not well understood, nor is it clear what unique functions, if any, miRNA family members possess. Here, we study evolutionary patterns of metazoan miRNAs, focusing on the targeting preferences of the let-7 and miR-10 families. These studies reveal hotspots for sequence evolution with implications for targeting and secondary structure. High-throughput screening for functional targets reveals that each miRNA represses sites with distinct features and regulates a large number of genes with cooperative function in regulatory networks. Unexpectedly, given the high degree of similarity, single-nucleotide changes grant miRNA family members with distinct targeting preferences. Together, our data suggest complex functional relationships among miRNA duplications, novel expression patterns, sequence change, and the acquisition of new targets.
Recent transcriptome data from yeast, worm, plants, and humans has shown that alternative polyadenylation (APA), a mechanism that enables expression of multiple 3′UTR isoforms for the same gene, is widespread in eukaryotic organisms. It is still poorly understood why metazoans require multiple 3′UTRs for the same gene, but accumulating evidence suggests that APA is largely regulated at a tissue-specific level. APA may direct combinatorial variation between cis-elements and microRNAs, perhaps to regulate gene expression in a tissue-specific manner. Apart from a few single gene anecdotes, this idea has not been systematically explored.
This dissertation research employs a systems biology approach to study the somatic tissue dynamics of APA and its impact on microRNA targeting networks in the small nematode C. elegans. In the first aim, tools were developed and applied to isolate and sequence mRNA from worm intestine and muscle tissues, which revealed pervasive tissue-specific APA correlated with microRNA regulation. The second aim provides genetic evidence that two worm genes use APA to escape repression by microRNAs in the body muscle. Finally, in aim three, mRNA from five additional somatic worm tissues was sequenced and their 3′ends mapped, allowing for an integrative study of APA and microRNA targeting dynamics in worms. Together, this work provides evidence that APA is a pervasive mechanism operating in somatic tissues of C. elegans with the potential to significantly rearrange their microRNA regulatory networks and precisely dose their gene expression.
Strategies are needed to improve repopulation of decellularized lung scaffolds with stromal and functional epithelial cells. We demonstrate that decellularized mouse lungs recellularized in a dynamic low fluid shear suspension bioreactor, termed the rotating wall vessel (RWV), contained more cells with decreased apoptosis, increased proliferation and enhanced levels of total RNA compared to static recellularization conditions. These results were observed with two relevant mouse cell types: bone marrow-derived mesenchymal stromal (stem) cells (MSCs) and alveolar type II cells (C10). In addition, MSCs cultured in decellularized lungs under static but not bioreactor conditions formed multilayered aggregates. Gene expression and immunohistochemical analyses suggested differentiation of MSCs into collagen I-producing fibroblast-like cells in the bioreactor, indicating enhanced potential for remodeling of the decellularized scaffold matrix. In conclusion, dynamic suspension culture is promising for enhancing repopulation of decellularized lungs, and could contribute to remodeling the extracellular matrix of the scaffolds with subsequent effects on differentiation and functionality of inoculated cells.
Deforestation in Myanmar has recently attracted much attention worldwide. This study examined spatio-temporal patterns of deforestation and forest carbon flux in Myanmar from 2001 to 2010 and environmental impacts at the regional scale using land products of the Moderate Resolution Imaging Spectroradiometer (MODIS). The results suggest that the total deforestation area in Myanmar was 21,178.8 km2, with an annual deforestation rate of 0.81%, and that the total forest carbon release was 20.06 million tons, with an annual rate of 0.37%. Mangrove forests had the highest deforestation and carbon release rates, and deciduous forests had both the largest deforestation area and largest amount of carbon release. During the study period, the south and southwestern regions of Myanmar, especially Ayeyarwady and Rakhine, were deforestation hotspots (i.e., the highest deforestation and carbon release rates occurred in these regions). Deforestation caused significant carbon release, reduced evapotranspiration (ET), and increased land surface temperatures (LSTs) in deforested areas in Myanmar during the study period. Constructive policy recommendations are put forward based on these research results.
This paper reviews how remotely sensed data have been used to understand the impact of urbanization on global environmental change. We describe how these studies can support the policy and science communities’ increasing need for detailed and up-to-date information on the multiple dimensions of cities, including their social, biological, physical, and infrastructural characteristics. Because the interactions between urban and surrounding areas are complex, a synoptic and spatial view offered from remote sensing is integral to measuring, modeling, and understanding these relationships. Here we focus on three themes in urban remote sensing science: mapping, indices, and modeling. For mapping we describe the data sources, methods, and limitations of mapping urban boundaries, land use and land cover, population, temperature, and air quality. Second, we described how spectral information is manipulated to create comparative biophysical, social, and spatial indices of the urban environment. Finally, we focus how the mapped information and indices are used as inputs or parameters in models that measure changes in climate, hydrology, land use, and economics.
This study examines the spatial and temporal patterns of the surface urban heat island (SUHI) intensity in the Phoenix metropolitan area and the relationship with land use land cover (LULC) change between 2000 and 2014. The objective is to identify specific regions in Phoenix that have been increasingly heated and cooled to further understand how LULC change influences the SUHI intensity. The data employed include MODerate-resolution Imaging Spectroradiometer (MODIS) land surface temperature (LST) 8-day composite June imagery, and classified LULC maps generated using 2000 and 2014 Landsat imagery. Results show that the regions that experienced the most significant LST changes during the study period are primarily on the outskirts of the Phoenix metropolitan area for both daytime and nighttime. The conversion to urban, residential, and impervious surfaces from all other LULC types has been identified as the primary cause of the UHI effect in Phoenix. Vegetation cover has been shown to significantly lower LST for both daytime and nighttime due to its strong cooling effect by producing more latent heat flux and less sensible heat flux. We suggest that urban planners, decision-makers, and city managers formulate new policies and regulations that encourage residential, commercial, and industrial developers to include more vegetation when planning new construction.
The urban heat island (UHI) phenomenon is a significant worldwide problem caused by rapid population growth and associated urbanization. The UHI effect exacerbates heat waves during the summer, increases energy and water consumption, and causes the high risk of heat-related morbidity and mortality. UHI mitigation efforts have increasingly relied on wisely designing the urban residential environment such as using high albedo rooftops, green rooftops, and planting trees and shrubs to provide canopy coverage and shading. Thus, strategically designed residential rooftops and their surrounding landscaping have the potential to translate into significant energy, long-term cost savings, and health benefits. Rooftop albedo, material, color, area, slope, height, aspect and nearby landscaping are factors that potentially contribute. To extract, derive, and analyze these rooftop parameters and outdoor landscaping information, high resolution optical satellite imagery, LIDAR (light detection and ranging) point clouds and thermal imagery are necessary. Using data from the City of Tempe AZ (a 2010 population of 160,000 people), we extracted residential rooftop footprints and rooftop configuration parameters from airborne LIDAR point clouds and QuickBird satellite imagery (2.4 m spatial resolution imagery). Those parameters were analyzed against surface temperature data from the MODIS/ASTER airborne simulator (MASTER). MASTER images provided fine resolution (7 m) surface temperature data for residential areas during daytime and night time. Utilizing these data, ordinary least squares (OLS) regression was used to evaluate the relationships between residential building rooftops and their surface temperature in urban environment. The results showed that daytime rooftop temperature was closely related to rooftop spectral attributes, aspect, slope, and surrounding trees. Night time temperature was only influenced by rooftop spectral attributes and slope.