Migration is a fundamental cellular behavior that plays an indispensable role in development and homeostasis, but can also contribute to pathology such as cancer metastasis. Due to its relevance to many aspects of human health, the ability to accurately measure cell migration is of broad interest, and numerous approaches have been developed. One of the most commonly employed approaches, because of its simplicity and throughput, is the exclusion zone assay in which cells are allowed to migrate into an initially cell-free region. A major drawback of this assay is that it relies on simply counting cells in the exclusion zone and therefore cannot distinguish the effects of proliferation from migration. We report here a simple modification to the exclusion zone migration assay that exclusively measures cell migration and is not affected by proliferation. This approach makes use of a lineage-tracing vital stain that is retained through cell generations and effectively reads out migration relative to the original, parental cell population. This modification is simple, robust, non-perturbing, and inexpensive. We validate the method in a panel of cell lines under conditions that inhibit or promote migration and demonstrate its use in normal and cancer cell lines as well as primary cells.
The goal of this thesis is to design, optimize and implement a device that can measure the oxygen consumption rate (OCR) of live single cells. A microfluidic device has been designed with the ability to rapidly seal and unseal microchambers containing individual cells and an extracellular optical oxygen sensor for measuring the OCR of live single cells. The device consists of two parts, one with the sensor in microwells (top half) and the other with channels and cells trapped in Pachinko-type micro-traps (bottom half). When the two parts of the device are placed together the wells enclose each cell. Oil is flown in through the channels of the device to produce isolated and sealed microchamber around each cell. Different fluids can be flowed in and out of the device, alternating with oil, to rapidly switch between sealed and unsealed microenvironment around each cell. A fluorescent ratiometric dual pH and oxygen sensor is placed in each well. The thesis focuses on measuring changes in the oxygen consumption rate of each cell within a well. Live and dead cells are identified using a fluorescent live/dead cell assay. Finally, the technology is designed to be scalable for high-throughput applications by controlling the flow rate of the system and increasing the cell array density.
Society is heavily dependent on a reliable electric supply; all infrastructure systems depend on electricity to operate. When the electric system fails, the impacts can be catastrophic (food spoilage, inoperable medical devices, lack of access to water, etc.). The social impacts, defined as the direct and indirect impacts on people, of power outages must be explored as the likelihood of power outages and blackouts are increasing. However, compared to other hazards, such as heat and flooding, the knowledge base on the impacts of power outages is relatively small. The purpose of this thesis is to identify what is currently known about the social impacts of power outages, identify where gaps in the literature exist, and deploy a survey to explore power outage experiences at the household level. This thesis is comprised of two chapters, a systematic literature review on the current knowledge of the social impacts of power outages and a multi-city survey focused on power outage experiences.
The first chapter comprised of a systematic literature review using a combined search of in Scopus which returned 762 candidate articles were identified that potentially explored the social impacts of power outages. However, after multiple filtering criteria were applied, only 45 articles met all criteria. Four themes were used to classify the literature, not exclusively, including modeling, social, technical, and other. Only papers that were classified as “social” - meaning they observed how people were affected by a power outage - or in combination with other categories were used within the review.
From the literature, populations of concern were identified, including minority demographics - specifically Blacks or African Americans, children, elderly, and rural populations. The most commonly reported health concerns were from those that rely on medical devices for chronic conditions and unsafe generator practices. Criminal activity was also reported to increase during prolonged power outages and can be mitigated by consistent messaging on where to receive assistance and when power will be restored. Providing financial assistance and resources such as food and water can reduce the crime rate temporarily, but the crime rate can be expected to increase once the relief expires. Authorities should expect looting to occur, especially in poorer areas, during prolonged power outages. Gaps in the literature were identified and future directions for research were provided.
The second chapter consists of a multi-city survey that targeted three major cities across the United States (Detroit, MI; Miami, FL; and Phoenix, AZ). The survey was disseminated through Amazon’s Mechanical Turk and hosted by Qualtrics. 896 participants from the three cities qualified to complete the full version of the survey. Three criteria had to be met for participants to complete the full survey including residing in one of the three target cities, living at their primary address for a majority of the year, and indicate they had experienced a power outage within the last five years.
Participants were asked questions regarding the number of outages experienced in the last five years, the length of their most recent and longest outage experienced, if they owned a generator, how they managed their longest power outage, if participants or anyone in their household relies on a medical device, the financial burden their power outage caused, and standard demographic- and income-related questions. Race was a significant variable that influenced the outage duration length but not frequency in Phoenix and Detroit. Income was not a significant variable associated with experiencing greater economic impacts, such as having thrown food away because of an outage and not receiving help during the longest outage. Additional assessments similar to this survey are needed to better understand household power outage experiences.
Findings from this thesis demonstrate traditional metrics used in vulnerability indices were not indicative of who experienced the greatest effects of power outages. Additionally, other factors that are not included in these indices, such as owning adaptive resources including medical devices and generators in Phoenix and Detroit, are factors in reducing negative outcomes. More research is needed on this topic to indicate which populations are more likely to experience factors that can influence positive or negative outage outcomes.
Access to air conditioned space is critical for protecting urban populations from the adverse effects of heat exposure. Yet there remains fairly limited knowledge of penetration of private (home air conditioning) and distribution of public (cooling centers and commercial space) cooled space across cities. Furthermore, the deployment of government-sponsored cooling centers is not based on the location of existing cooling resources (residential air conditioning and air conditioned public space), raising questions of the equitability of access to heat refuges.
Using Los Angeles County, California and Maricopa County, Arizona (whose county seat is Phoenix) we explore the distribution of private and public cooling resources and access inequities at the household level. We do this by evaluating the presence of in-home air conditioning and developing a walking-based accessibility measure to air conditioned public space using a combined cumulative opportunities-gravity approach. We find significant inequities in the distribution of residential air conditioning across both regions which are largely attributable to building age and inter/intra-regional climate differences. There are also regional disparities in walkable access to public cooled space.
At average walking speeds, we find that official cooling centers are only accessible to a small fraction of households (3% in Los Angeles, 2% in Maricopa) while a significantly higher number of households (80% in Los Angeles, 39% in Maricopa) have access to at least one other type of public cooling resource which includes libraries and commercial establishments. Aggregated to a neighborhood level, we find that there are areas within each region where access to cooled space (either public or private) is limited which may increase the health risks associated with heat.
Hydrophobic platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorophenyl)-porphyrin (PtTFPP) was physically incorporated into micelles formed from poly(ε-caprolactone)-block-poly(ethylene glycol) to enable the application of PtTFPP in aqueous solution. Micelles were characterized using dynamic light scattering (DLS) and atomic force microscopy (AFM) to show an average diameter of about 140 nm. PtTFPP showed higher quantum efficiency in micellar solution than in tetrahydrofuran (THF) and dichloromethane (CH2Cl2). PtTFPP in micelles also exhibited higher photostability than that of PtTFPP suspended in water. PtTFPP in micelles exhibited good oxygen sensitivity and response time. This study provided an efficient approach to enable the application of hydrophobic oxygen sensors in a biological environment.
With the successful development of organic/polymeric light emitting diodes, many organic and polymeric fluorophores with high quantum efficiencies and optical stability were synthesized. However, most of these materials which have excellent optical properties are insoluble in water, limiting their applications in biological fields. Herein, we used micelles formed from an amino-group-containing poly(ε-caprolactone)-block-poly(ethylene glycol) (PCL-b-PEG-NH2) to incorporate a hydrophobic blue emitter oligofluorene (OF) to enable its application in biological conditions. Although OF is completely insoluble in water, it was successfully transferred into aqueous solutions with a good retention of its photophysical properties. OF exhibited a high quantum efficiency of 0.84 in a typical organic solvent of tetrahydrofuran (THF). In addition, OF also showed a good quantum efficiency of 0.46 after being encapsulated into micelles. Two cells lines, human glioblastoma (U87MG) and esophagus premalignant (CP-A), were used to study the cellular internalization of the OF incorporated micelles. Results showed that the hydrophobic OF was located in the cytoplasm, which was confirmed by co-staining the cells with nucleic acid specific SYTO 9, lysosome specific LysoTracker Red®, and mitochondria specific MitoTracker Red. MTT assay indicated non-toxicity of the OF-incorporated micelles. This study will broaden the application of hydrophobic functional organic compounds, oligomers, and polymers with good optical properties to enable their applications in biological research fields.
Single-cell studies of phenotypic heterogeneity reveal more information about pathogenic processes than conventional bulk-cell analysis methods. By enabling high-resolution structural and functional imaging, a single-cell three-dimensional (3D) imaging system can be used to study basic biological processes and to diagnose diseases such as cancer at an early stage. One mechanism that such systems apply to accomplish 3D imaging is rotation of a single cell about a fixed axis. However, many cell rotation mechanisms require intricate and tedious microfabrication, or fail to provide a suitable environment for living cells. To address these and related challenges, we applied numerical simulation methods to design new microfluidic chambers capable of generating fluidic microvortices to rotate suspended cells. We then compared several microfluidic chip designs experimentally in terms of: (1) their ability to rotate biological cells in a stable and precise manner; and (2) their suitability, from a geometric standpoint, for microscopic cell imaging. We selected a design that incorporates a trapezoidal side chamber connected to a main flow channel because it provided well-controlled circulation and met imaging requirements. Micro particle-image velocimetry (micro-PIV) was used to provide a detailed characterization of flows in the new design. Simulated and experimental results demonstrate that a trapezoidal side chamber represents a viable option for accomplishing controlled single cell rotation. Further, agreement between experimental and simulated results confirms that numerical simulation is an effective method for chamber design.
Maricopa County is the home of the Phoenix metropolitan area, an expansive city with serious air quality concerns. To ameliorate air quality in the county, the Maricopa County Air Quality Department developed a website and mobile application called "Clean Air Make More" as a means of outreach and engagement. In doing this, the county has found a way to engender a bilateral relationship between individuals and their government agency. This study analyzes the effectiveness of Clean Air Make More in establishing this relationship and engaging the community in efforts to improve air quality. It concludes that the design of the application effectively meets user needs, but marketing efforts should target populations disposed to taking action regarding air quality.
Background:
Environmental heat exposure is a public health concern. The impacts of environmental heat on mortality and morbidity at the population scale are well documented, but little is known about specific exposures that individuals experience.
Objectives:
The first objective of this work was to catalyze discussion of the role of personal heat exposure information in research and risk assessment. The second objective was to provide guidance regarding the operationalization of personal heat exposure research methods.
Discussion:
We define personal heat exposure as realized contact between a person and an indoor or outdoor environment that poses a risk of increases in body core temperature and/or perceived discomfort. Personal heat exposure can be measured directly with wearable monitors or estimated indirectly through the combination of time–activity and meteorological data sets. Complementary information to understand individual-scale drivers of behavior, susceptibility, and health and comfort outcomes can be collected from additional monitors, surveys, interviews, ethnographic approaches, and additional social and health data sets. Personal exposure research can help reveal the extent of exposure misclassification that occurs when individual exposure to heat is estimated using ambient temperature measured at fixed sites and can provide insights for epidemiological risk assessment concerning extreme heat.
Conclusions:
Personal heat exposure research provides more valid and precise insights into how often people encounter heat conditions and when, where, to whom, and why these encounters occur. Published literature on personal heat exposure is limited to date, but existing studies point to opportunities to inform public health practice regarding extreme heat, particularly where fine-scale precision is needed to reduce health consequences of heat exposure.
This report summarizes the proceedings of the workshop focusing primarily on two sessions: the first related to social vulnerability mapping and the second related to the identification and prioritization of interventions necessary to address the impacts of climate-sensitive hazards.