There has been growing scholarly attention to the fact that most global health projects/research are disproportionately focused on specific locales resulting in temporary restricted interventions. This approach is, in some significant sense a 'betrayal' of the very idea of global health. This study asks how Seattle's various global health institutions and organizations approach Global health in Kenya. I hypothesize that most global health projects promote the production of inequity, unequal and uncaring exchanges between donor institutions and the recipients. I analyze the socio-historical, economic and political histories and ties between Kenya and global health institutions in Seattle drawing on political-economy analysis and feminist care ethical analysis. I also thematically code and analyze ethnographic interviews to investigate the relationship between the responses and the main research topic. I expect to find that global health projects may not necessarily achieve their intended goals because they do not fully consider the four phases of care, which involve attentiveness, responsibility, competence and responsiveness. Understanding the factors influencing the production of unequal and uncaring exchanges in global health is essential as it will impact how global health projects should be conducted.

Ion mobility mass spectrometry is a powerful technique for analyzing proteins and protein complexes in the gas phase. Ions are generated using nanoelectrospray ionization from aqueous, buffered solutions containing the protein or protein complex of interest. The resulting ions are native-like; their structures have low, narrow charge-state distributions and resemble those in the condensed-phase. There is an interest in probing different charge states of protein and protein complexes. High charge-state distributions of proteins and protein complexes, for example, can be seen after addition of ‘supercharging’ reagents, e.g., sulfolane or m-nitrobenzyl alcohol, to the electrospray buffer. Several mechanisms have been proposed to explain the mechanism of supercharging. Here, we used ion mobility mass spectrometry to probe the charge-state distributions and collision cross sections of a large set of native-like protein and protein complex ions, with masses ranging from 6–468 kDa, generated from solutions containing 1% m-nitrobenzyl alcohol. We determined the extent of supercharging by calculating the percent increase in highest charge state observed between supercharged and non-supercharged cations. We observed a large range of percent increases in highest charge state but in general, there is evidence that small proteins (<100 kDa) exhibited a greater increase than larger proteins in these experiments. Upon comparison of these results to those for ions generated from sulfolane-containing solutions, we found that the maximum extent of supercharging observed with m-nitrobenzyl alcohol was less than that observed when sulfolane was used. The most supercharged cations generated with m-nitrobenzyl alcohol had up to a 96% increase in collision cross section compared to ions generated without supercharging agents, but the differences in collision cross sections with and without supercharging were less than 2% for large protein complexes (> 100 kDa). These results suggest that ions supercharged using m-nitrobenzyl alcohol may retain their structures better than those supercharged using sulfolane.

The 21st century has seen the emergence and re-emergence of novel infectious diseases. Following in the footsteps of its communicable predecessors, the recent emergence in Saudi Arabia of a SARS-like novel coronavirus, or Middle East respiratory syndrome (MERS-CoV), since 2012 has weaved narratives of globally interdependent economic, political, and social intimacies. I will examine the contrasting narratives that reflect investigations and responses to this novel threat from Middle Eastern, European, and North American media networks, the WHO, the global biomedical community, and the Saudi Arabian Ministry of Health through several intersecting discourses: intellectual property rights and viral sovereignty; the deliberation on the taxonomy of the novel coronavirus; and geopolitical differences in information sharing and biosecurity in the context of emergent 21st century epidemics. Using Priscilla Wald’s concept of outbreak narratives and the imaginative hold of myths as a framework, I will explore how the stories on the global effort to investigate and respond to the MERS outbreaks continue to be revealed, repackaged, and transformed as experts around the world scramble to uncover its epidemiological pathways. I argue that the consideration of these counter-narratives are critical for rethinking underlying, potentially harmful, factors beyond dominant epidemiological and biomedical discourses surrounding the MERS epidemics. Such considerations may serve as a microcosm for biopolitical barriers in the response to past, present, and future emergent diseases. These counter-narratives should be regarded in light of uncertainties that still cloud the etiology and transmission of the virus which has since infected 243 people and claimed 93 lives in 11 countries in Europe, the Middle East, and Southeast Asia.

The somatic mutation theory of aging states that aging is the result of accumulated DNA mutations in somatic cells, which lead to loss of physiological integrity through tissue and organ level dysfunction. In mouse model systems, decreases in lifespan through high mutation rates have been documented by creating mutator phenotypes in these organisms. Mutator phenotypes are marked by increased cellular mutation rates, and are known drivers of tumor development in human cancers. Mutator-driven adaptations occur in natural microbial populations. Mutators represent a unique opportunity to test the somatic mutation accumulation theory of aging due to their sustained high mutation rates. Nutritional status, particularly carbohydrate metabolism, influences lifespan in numerous organisms. In yeast, which preferentially ferments glucose to ethanol before switching to respiratory metabolism upon glucose depletion, the induction of respiratory metabolism is associated with increased lifespan. This study examines the effects of high mutation rate on yeast replicative lifespan and whether induction of respiratory metabolism is sufficient to change lifespan in organisms with increased mutation load. To do this, yeast defective for DNA replication proofreading, mismatch repair, and base-excision repair were grown under dietary restriction (reduced glucose) and in the presence of non-fermentable carbon sources. Then, mutation rates and replicative lifespan were assayed. Our results show that increased mutation rates correlate with decreased lifespan in strong mutators and that this decrease in lifespan can be partially suppressed through growth on limited glucose and non-fermentable media. These findings suggest that the metabolic state of the cell modulates the severity of mutator phenotypes, which has important implications for the role that mutators play in carcinogenesis.

This paper analyzes the juxtaposing narratives between various documents and media portrayals from mining corporations, the Peruvian government, and the affected indigenous populations, focusing on how many of the more dominant narratives can often be misleading in reflecting the serious health effects from mining. Peru’s mining industry significantly influences the health of surrounding indigenous communities due to high levels of air and water pollution. Unfortunately, there is not a great deal of sincere concern for the wellbeing of these communities from either the corporations exploiting their land or the Peruvian government. Despite their efforts to express their suffering through protests, indigenous communities have seen few results. The mining industry and its detrimental health effects are often overlooked, and therefore many of these dominant corporate narratives and our own capitalist desires are all that we have to shape our understanding of how mining industries operate in other countries, such as Peru. However, the subaltern counter-narratives from indigenous populations concerning their health and rights can more accurately shape our understanding of the severity of these health effects caused by the mining industry.

To address the growing international attention on the high rates of obesity in the Pacific Islands, this project examines the discursive patterns that play a role in both fueling and quelling the outbreak of obesity among the Samoan population. I will provide a historical account of the diet and lifestyle during the pre- and post-colonialism periods in American Samoa. I am especially interested in tracing the racialization of food and the differences in perceiving food. I draw on theorist Michel Foucault’s interpretation of biopower in  to further illuminate the narratives that perpetuate the issues of power inequality, colonialism and the racialization of obesity towards the Samoan people, and what exactly defines “health” and “wellness”. The sources of these narratives are extracted from everday popular media, news articles, and even film or artwork. The project also uncovers the various counter narratives from Samoan Americans in the local Seattle community as well as Pacific Island authors and educators to shed light on work counteracting obesity in the community, the significance of cultural preservation and indigenizing health and incorporating de-colonial movements. This leads into the movement towards individualized risk management awhile not losing sight of community and relationship. The implications of this analysis lead us to consider the correlation between discourse and body governance in the context of a growing global health and food industry.

The purpose of this research is to investigate hidden health narratives on campus regarding the use of tobacco products to contextualize the legitimacy of smoke-free and tobacco-free university initiatives. This academic endeavor will also reveal the structural and institutionalized violence of the tobacco industry which targets marginalized communities to entice the uptake of regular smoking habits. An analysis of biopolitical power, structural violence, and social theory will help to reveal the hidden narratives of these populations, twice-targeted by the social stigmatization of tobacco use and the manipulative advertising of the tobacco industry. By exposing, analyzing, and deconstructing patterns of profit-driven tobacco marketing directed at student demographics, it is possible to examine geographies of blame regarding tobacco use, impact of tobacco use on non-smoking populations, and the misleading agendas behind the dominant narratives of tobacco industry advertising. By identifying and bringing to light these messages, it is possible to legitimize the anti-smoking and anti-tobacco movements happening at the University of Washington, and U.S. universities at large, with a critical analysis of the health risks involved.

Aging research depends on the use of model organisms to test hypotheses about the biological mechanisms common to many organisms. One commonly used model organism is the roundworm Caenorhabditis elegans. Unlike homeothermic mammals, C. elegans are poikilotherms, and as a result of a variable internal temperature, they also exhibit variable longevity based on the ambient temperature. Under standard laboratory conditions, C. elegans are commonly grown in incubators ranging between 15°C and 25°C, and lifespan varies inversely with temperature; worms live longer at colder temperatures. Our recent data show that the relationship between temperature and aging has a genetic component. By measuring the lifespan of worm strains with mutations in genes found in established aging pathways, we have identified mutations that have opposing effects on lifespan at different temperatures. For example, utilizing lifespan assays I have conducted, we have found worm strains that live longer than controls at high or low temperature, but not both, as well as strains that are short-lived at specific temperatures. These experiments suggest there may be overlapping but distinct mechanisms driving aging at different temperatures, and we are developing hypotheses to explain the temperature-dependent mechanisms of aging. Thus far, our data are consistent with the hypothesis that genes that contribute to protein homeostasis promote longevity at higher temperatures, whereas genes that limit effects of toxic compounds (both internally and externally produced) promote longevity at low temperatures. Understanding the temperature-dependent mechanisms of aging in C. elegans will advance the aging research field by extending knowledge on a widely used model organism and by testing how the mechanisms of aging and disease may be conserved in homeotherms. We hope our results will suggest how and why many genetic longevity pathways are conserved from worms to mammals despite drastically different life histories.

Tyrosinemia Type II is a genetic disorder caused by a mutation in the enzyme tyrosine aminotransferase (TAT), which leads to an elevated level of tyrosine in the blood. If untreated, individuals with tyrosinemia develop life-threatening clinical symptoms that include an increased tendency to bleed, damage to the nervous system, damage to the eyes, and organ failure. Unexpectedly, mutation of TAT in the nematode Caenorhabditis elegans results in animals that are long lived in comparison with wild-type (WT) worms. This study aims to elucidate the cellular mechanism and workings of TAT in the mitochondria, its role in the pathology of the disease, and its possible links with aging in C. elegans. Thus far, we have developed a simple method to measure the CO2 output of the worms and shown that the TAT mutant’s metabolism is about 50% compared to WT. This significant decrease in CO2 output was matched comparable to a strain deficient for mitochondrial Cytochrome C Oxidase, which served as a positive control. Cytochrome C Oxidase, or Electron Transport Chain Complex IV, is one of the three complexes that create the proton gradient inside the mitochondrial matrix upon accepting electrons. Notably, mutation of the gene acting downstream to TAT in the tyrosine degradation pathway, which causes Tyrosinemia Type III in people, or any other mutation in tyrosine metabolism that we have tested, causes no significant change in CO2 output of C. elegans. Furthermore, through our preliminary metabolomics assay we have identified three metabolites that are accumulated to extremely high levels in comparison to positive and negative controls. The future steps will include determining whether these metabolites play a direct role in the effects of TAT mutation on mitochondrial function and aging.

This project examines the role of gender inequality in HIV/AIDS and how this inequality perpetuates the susceptibility of women to contract the virus. Focusing on Ghana and Brazil, I explore how culture and society shape an imbalance that potentially increases risks to women in becoming sick through analysis of the relationship between social determinants of health, risk factors and vulnerability. I argue that preventative measures intended to fight against HIV/AIDS have failed to consider the social circumstances that place women at greater risk than men. In my analysis, I draw on and interpret Michel Foucault’s concept of biopolitics using feministic critique and gendered perspectives. I reference anecdotes from Stephanie Nolen’s “28: Stories of AIDS in Africa” to present personalized perspectives of the painfully multifaceted reality of the HIV/AIDS outbreak, while Silvana Paternostro’s personal accounts of sexual culture in Brazil in “In the Land of God and Man: Confronting Sexual Culture in Latin America,” serve as a framework for my examination of gendered social constructs in Brazil. I examine gender relations and the dynamic of marital relationships that often disproportionately allow the male greater power over the female, and how a male dominant ideology is reinforced by politics, culture and social norm. Distinguishing between gender and sex, I look at how gender inequality, the inequality of opportunities and power that follow being a certain sex, may expose mothers, daughters, sisters, and other women at risk in ways we may not have imagined.

The mammalian Target of Rapamycin (mTOR) is a protein kinase that integrates signaling from growth factors and nutrient availability to regulate metabolism and cell growth. Reducing mTOR signaling with rapamycin extends lifespan in several model organisms such as yeast, flies, and mice, and rescues virtually all symptoms in a mouse model of Leigh syndrome, a neurodegenerative disorder that causes loss of motor function and early death. Previous observations from our laboratory suggest that rapamycin induces changes in the metabolism of mice modeling Leigh syndrome, favoring fatty and amino acid catabolism over glycolysis. We believe this shift in metabolism is, at least in part, responsible for the beneficial effects of rapamycin in normative aging. In order to test this hypothesis, we are examining the effects of daily rapamycin injections in 4, 12, and 22 months old mice. We performed baseline measurements of serum metabolites, forelimb strength, voluntary running, gait and balance, body composition, blood glucose, and hemoglobin A1c, and are currently administering 8mg/kg of rapamycin daily via intra-peritoneal injections for 60 days. During this period we are monitoring body weight and food intake daily, as well as blood glucose every 15 days. At the end of this period we will measure body composition, gait and balance, forelimb strength, and voluntary running again, while maintaining the mice under treatment with rapamycin for an additional 30 days. Finally, we will sacrifice the mice and analyze serum, liver, brain, adipose, and other tissues for the activation of metabolic pathways, metabolite composition, and signs of pathology. We expect rapamycin to improve most parameters studied and profoundly alter the metabolic profile of mice at all three ages. These results will ultimately allow us to investigate how modulating metabolic pathways can increase healthspan in mice and provide insight on how to promote healthy aging in humans.

Leprosy, also known as Hansen’s disease, is a deteriorative skin condition caused by chronic infections of the bacteria Mycobacterium leprae and Mycobacterium lepromatosis. Leprosy was once believed to be highly contagious, so patients with leprosy were quarantined and isolated to prevent further contamination within the population. In Korea, patients with leprosy were quarantined on Sorok Island where some patients received treatment for leprosy with the antibiotic, 4,4'-diaminodiphenylsulfone (DDS). Data collected on patients receiving DDS indicated that the average lifespans of the patients were longer than the average lifespans of the general Korean population suggesting that DDS may impact aging. Consistent with this idea, DDS has been reported to extend lifespan in the nematode Caenorhabditis elegans. We have confirmed these results in C. elegans, and the goal of this project is to identify the specific mechanisms and pathways through which DDS is able to extend lifespan. Our unpublished data support the model that one of the mechanisms underlying DDS’s effect on aging in C. elegans is activation of AMP- activated protein kinase (AMPK). We are currently working to understand how DDS activates AMPK, as well as the interaction between DDS and other known longevity pathways in C. elegans, such as dietary restriction and the hypoxic response. I am analyzing the mechanisms through which DDS affects aging by performing lifespan analysis on C. elegans and its mutants. By analyzing the effects of DDS on aging, we can get a better understanding of the mechanisms that control aging to eventually comprehend the pathways through which certain aging diseases, like Alzheimer's act.

The Rieske iron-sulfur protein (Rip1) is a nuclear encoded protein that assembles into the cytochrome bc1 complex of the electron transport chain in budding yeast, Saccharomyces cerevisiae. The iron-sulfur cluster bound by this protein facilitates the flow of electrons during cellular respiration in the inner membrane of the mitochondrial matrix. In the nematode worm, Caenorhabditis elegans, a mutation in a conserved residue of the Rieske iron-sulfur protein has been found that delays development and extends the lifespan of this organism. Mutagenesis experiments from our laboratory have identified suppressors of this C. elegans developmental phenotype that are due to secondary mutations in conserved residues within a flexible hinge region of the Rieske iron-sulfur protein. The primary mutation and secondary suppressing mutations were introduced to the yeast Rip1 gene through site directed PCR mutagenesis. Transgenic yeast have been created expressing the mutant forms of Rip1 homologous to those identified in C. elegans. The next stages of my project will entail the characterization of these mutations in yeast. I plan to assay the replicative lifespan, growth rate, respiratory efficiency, as well as sensitivity to variance in temperature, reactive oxygen species and carbon source in an effort to understand how this mutant’s lifespan extension fits into the larger picture of known aging pathways. The extrapolation of the mechanisms found in these model organisms serve as the basis for understanding our own complex aging process.

Parkinson’s Disease (PD) is the second most common neurodegenerative disorder in the world, affecting nearly 7 million people globally and more than 1 million people in the U.S. Symptoms of PD typically include tremors, rigidity, postural instability and slowness of movement. These motor control deficiencies can be attributed to a decrease in the expression of the neurotransmitter dopamine and the loss of many dopaminergic neurons in the substantia nigra of the brain. To study PD a mouse model is used in which C57BL/6 mice are administered 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). MPTP is a neurotoxin that inhibits complex I activity in the mitochondrial electron transport chain. Mice treated with MPTP exhibit PD-like symptoms, including the loss of dopamine producing neurons and physical motor control deficiencies analogous to PD. Specifically, we are interested in utilizing the MPTP mouse model to understand a mechanism by which the drug rapamycin might prevent or treat symptoms of PD. Rapamycin inhibits signaling of the mechanistic target of rapamycin (mTOR) pathway, which is a key regulator of many functions, including cell growth, cell proliferation, cell motility, cell survival, protein synthesis, and transcription. Previous research by our lab has shown that rapamycin dramatically attenuates disease progression in a mouse model of mitochondrial complex I deficiency. Because PD patients also exhibit defects in complex I, we believe that rapamycin may alleviate Parkinson’s disease progression by a mechanism similar to the effects we have seen. Currently we are administering high doses of encapsulated rapamycin in mouse diet during the three weeks prior to MPTP exposure. The expression of PD-like biomarkers in rapamycin fed mice will be measured through immunohistochemistry and western blot assays and compared to a control fed group. We hypothesize that pre-treatment with high doses of rapamycin in the diet will have robustly beneficial effects in MPTP mice.

 The Gram negative, opportunistically pathogenic Gammaproteobacterium Pseudomonas aeruginosa has a propensity to form biofilms. Biofilms are surface associated or aggregated bacterial communities, embedded in a self-derived, extracellular matrix (EPS). One major component of the P. aeruginosa biofilm EPS is the polysaccharide Psl. Psl plays a structural and protective role in P. aeruginosa biofilms by mediating attachment to abiotic surfaces and mammalian cells, conferring protection from components of the host immune response and a variety of antimicrobials, as well as promoting cell-cell adhesion. More recently, Psl has also been shown to function as a cell-cell signaling molecule that activates two diguanylate cyclases through an unknown mechanism, which results in an increase in intracellular levels of c-di-GMP. The secondary messenger c-di-GMP is central to the P. aeruginosa virulence regulatory network, and promotes biofilm formation, in part, by inducing the production of EPS components, including Psl. Thus, presence of environmental Psl appears to act as a feed-forward signaling molecule that enhances biofilm formation. In order to further understand this function, this investigation will employ mini-Tn5 mutagenesis to define the signal transduction pathway(s) responsible for activating the diguanylate cyclases through a novel feed-forward mechanism of Psl-dependent signaling in P. aeruginosa.

 The mitochondria are cell organelles that perform important cellular functions such as energy production, regulation of the cell cycle, and cell death. Therefore, proper mitochondrial function is critical for healthy aging of an organism. Paradoxically, attenuation of mitochondrial function in the nematode worm Caenorhabditis elegans has been observed to extend lifespan. It has been proposed that induction of the mitochondrial unfolded protein response (UPRmt) is responsible for this enhanced longevity. The UPRmt is a response to proteotoxic stress in the mitochondria that results in induction of nuclear-encoded mitochondrial chaperones, proteins that stabilize and fold misfolded and aggregated proteins. However, the mechanism(s) by which proteotoxic stress is recognized in the mitochondria and signaled to the nucleus, resulting in the UPRmt response, remain largely unknown. To further understand the UPRmt pathway, we carried out a genome-wide RNAi screen to search for RNAi clones that specifically reduce UPRmt signaling under conditions of mitochondrial stress. To do this, we used a mutation in the electron transport chain (ETC) gene isp-1 to cause constitutive activation of the UPRmt GFP reporter, hsp-6p::gfp and screened for RNAi clones that reduced GFP fluorescence. We expect that positive hits in our screen will target signaling components of the UPRmt. In total, we identified approximately 60 RNAi clones that reduce the UPRmt response in isp-1(qm150) worms. Currently, we are re-testing positive hits after sequence validation and determining if the targeted genes specifically signal the UPRmt or non-specifically affect expression of GFP reporters. Once we identify genes that are specific to the UPRmt, we plan to test the role of these genes in mitochondrial function and lifespan. A better understanding of the UPRmt pathway could provide significant insight into mitochondrial biology, which impacts healthy aging and several age-related diseases.

Mitochondria, the energy producing organelles in eukaryotic cells, play a critical role in cell metabolism and regulation. Interestingly, mitochondrial dysfunction promotes lifespan extension in Caenorhabditis elegans. One proposed mechanism is that the induction of mitochondrial unfolded protein response (UPR^mt) increases the expression of mitochondrial chaperones, nuclear-encoded enzymes that assist in folding of misfolded or aggregated proteins, to maintain mitochondrial protein homeostasis. In an effort to elucidate genes involved in the UPR^mt pathway, we performed a genome-wide RNAi screen in transgenic animals expressing mitochondrial chaperone GFP reporter hsp-6_pr::gfp. Unexpectedly, we found that knockdown of genes involved in the pentose phosphate pathway (PPP) activates hsp-6_pr::gfp expression. The PPP is a metabolic shunt off of glycolysis that produces NADPH, a reducing equivalent used for glutathione reduction and lipid metabolism, and ribose 5-phosphate for nucleotide synthesis. In addition, knockdown of transaldolase, a gene in the reductive phase of the PPP, increases the lifespan of C. elegans. Through lifespan epistasis analysis, we found that transaldolase deficiency promotes longevity by a mechanism that is genetically distinct from known longevity factors such as the insulin-like/IGF-1 target (DAF-16), the hypoxia inducible factor-1 (HIF-1), AMP kinase (AAK-2), the UPR^mt transcription factor (ATFS-1), or the germline signaling (GLP-1). This suggests that the PPP inhibition promotes longevity through an uncharacterized pathway that may respond to mitochondrial dysfunction. In addition, we found that transaldolase knockdown alters expression of fatty acid desaturase, an enzyme that oxidizes fatty acid chains. We hypothesize that dysregulation of lipid metabolism by inhibition of the PPP may cause a defect in mitochondrial lipid composition, which may increase mitochondrial proteotoxic stress, inducing the UPR^mt. Understanding the genetics behind the UPR^mt and the PPP’s role in aging has enormous benefits to human health as both the PPP and mitochondrial dysfunction are highly correlated with many age-related diseases including cancer and neurodegenerative diseases.

This project examines the ethical and social frames which facilitate environmental conditions that fuel the rampage of tuberculosis in India. Three key components of the basic biomedical outbreak narrative that are often overlooked will be brought forth to attention: the tuberculosis outbreak as more than just a bacterial spread, social constructs which shape the bacteria’s accessibility to victims, and socioeconomic determinants of treatment (and lack thereof). India accounts for approximately one-fifth of global tuberculosis occurrences, and with over 2 million cases in the country, tuberculosis is a continuously growing health concern, posing a major threat to the well-being of the population. Due to India's developing health care system, pollution, and poverty, these conditions amplify the ease of transmissibility and rate of infection. As treatment regimens are abandoned due to social stigma, medical side effects, and overwhelming price tags, the bacterium which causes TB (mycobacterium tuberculosis), tends to mutate and become drug-resistant to varying degrees; increasing the difficulty of containment and scientific development of effective medicines. Furthermore, the capitalist structure of the pharmaceutical industry contributes to the financial challenges which prevent the eradication of tuberculosis. Drawing on the theory of biopolitics while using sources of statistical information and analytical discourse,  my presentation seeks to explore various properties of the disease and discover how the complex interconnectedness of these underlying issues attribute to forms of structural violence, perpetuating the growth of tuberculosis in India.

One of the goals of modern longevity research is to find pharmacological means to increase lifespan and/or healthspan without deleterious physiological effects. Berberine is a yellow, bitter-tasting natural alkaloid with a long history in Eastern Asian medicinal use in treatments of a variety of age-associated diseases and infections. In human cell culture models, berberine treatment increases AMPK activity, resulting in delayed senescence through inhibition of the mTOR/S6 pathway. Here, we use the budding yeast Saccharomyces cerevisiae and the nematode Caenorhabditis elegans to investigate the effects of berberine on healthspan and lifespan. For the yeast S. cerevisiae, growth rates of haploid and diploid yeast cells, as well as chronological (a measure of population viability through time) and replicative (a measure of mitotic potential) yeast aging paradigms were assayed in the presence of concentrations of berberine ranging from 0.1 - 100.0 ng/uL. For C. elegans, organismal lifespan as well as various healthspan measures are being analyzed. Currently, we have found that concentrations of berberine greater than 10 ng/uL in haploids are sufficient to inhibit cell growth and lower concentrations (0.1-10 ng/uL) result in increased yeast viability when shifted to respiratory metabolism, a means of extending lifespan in yeast. Our lifespan studies are ongoing. Our experiments will provide valuable insights into the role of this long-used medicinal substance on cell aging and health using two important longevity models.

One goal of aging research is to identify genes that modulate aging, in order to eventually develop therapies that delay the onset of age-associated diseases. In particular, genes that control mitochondrial function appear to play an important role in aging. Although severe mitochondrial dysfunction causes childhood diseases such as Leigh syndrome, there is growing evidence that moderate mitochondrial stress can extend lifespan in yeast, worms, fruit flies, and even mice. In the nematode Caenorhabditis elegans, a long-lived mitochondrial mutant called isp-1(qm150) has a defect in the Rieske iron-sulfur protein. The Rieske iron–sulfur protein is a primary electron acceptor located in complex III of the mitochondrial electron transportation chain. Complete RNAi knockdown of isp-1 results in larval arrest, while partial knockdown extends lifespan, further supporting the idea that milder forms of mitochondrial stress can increase lifespan while severe mitochondrial stress is detrimental. We are conducting a mutagenesis screen to identify genetic suppressors of the larval developmental arrest induced by isp-1(RNAi). Through exposure to EMS/ENU mutagen, wild-type worms will have random mutations generated throughout their genomes. These mutants will then be screened for the ability to develop on isp-1(RNAi) . We speculate the novel suppressor mutations from the screen may affect proteins that interact with ISP-1 or cause mutations that bypass the need for ISP-1 in the mitochondria. Identification of such suppressors will help to clarify the mechanisms linking mutation of isp-1 to development and aging. The isp-1 gene and its function in the electron transport chain are conserved in humans. Therefore these studies may be relevant for human aging and disease.

Recent developments in research on Human Papillomavirus (HPV) have allowed for an increase of strategies that prevent the onset of cervical cancer within communities of little economic privilege. This project will compare preventative measures taken in India and those currently called for within the U.S., primarily examining how cultural and economic limitations are taken into consideration. As a “space of negation and the socially dead, produced by expropriations and alienations in and outside European nation-states” necropolis or low and middle income countries have historically been perceived as a necessity in order to maintain the priorities set by biopolis. However, organizations such as PATH are beginning to look into more cooperative methods. I will analyze the onset of preventative practices that promote self sufficiency in a format that maintains economic functionality and decreases potential dependency for aid. One significant limitation however is that western corporations still hold much of the control regarding economic leeway of vaccination use and price on a global scale. In this project I explore alternative methods that advocate information as an alternative or supplemental method for prevention. This project will continue by examining how circumstances in low economic environments are taken into consideration through efforts of vaccination or treatment. This includes the incorporation of cultural awareness, provision of knowledge and communication within the community, and how this leads to a stronger awareness of what limitations remain.

The Kaeberlein laboratory uses the budding yeast Saccharomyces cerevisiae as a model organism in the study of aging. S. cerevisiae is a serviceable model organism for this task because its lifespan is relatively short and simple to assess. Furthermore, many genetic tools are available to study the effect of individual genes on lifespan. Two experimental paradigms have been extensively used to quantify lifespan in yeast: chronological and replicative. In the chronological lifespan model, cells are maintained in a non-dividing state and viability is determined over time by assessing the ability of cells to re-enter the cell cycle upon exposure to growth promoting conditions. In the replicative lifespan (RLS) model, mitotically active cells are incubated until they divide and produce newly budded daughter cells, which are then micro-dissected away from their mother cells and counted. This process is repeated until daughter cells are no longer produced by the mother cell, and the total number of daughters produced prior to irreversible cell cycle arrest is quantified. After analyzing numerous strains of S. cerevisiae, many different genes and environmental conditions that modulate replicative lifespan have been identified. Importantly, many longevity modifiers identified in yeast have been found to be conserved in higher eukaryotes.

Aging is a degenerative process characterized by progressive deterioration of cellular components resulting in mortality. The nematode Caenorhabditis elegans has been used extensively to study the biology of aging, and several determinants of C. elegans longevity are conserved in mammals. The specific mechanisms of longevity in C. elegans have not been completely identified, and many pathways are poorly understood. Dietary restriction (DR) is one promising pro-longevity intervention, though its mechanism is still poorly defined. DR, a reduction in caloric intake in absence of malnutrition, extends lifespan in C. elegans and in a wide range of other species including yeast, flies, mice, and primates. To investigate the genetic determinants of the response to DR, we are performing a genome-wide screen using the Vidal RNA-interference (RNAi) Library, which contains 11,511 unique RNAi clones. The C. elegans are fed E. coli that express dsDNA that, when ingested, form ribonucleic-protein complexes that bind to and facilitate cleavage of target mRNAs; effectively preventing expression of specific genes. By knocking down specific genes within the C. elegans genome through RNAi, we are able to observe extension or reduction in longevity in response to DR. We have screened over 8,143 RNAi clones; 71 of which reduce lifespan under DR conditions and 85 of which increase lifespan. We are currently following up on our data by verifying the effect of these RNAi clones on lifespan under both control and DR conditions. We hope to verify how significantly these genes are affected by DR and follow up on those specific pathways to further investigate the interaction between DR and other potential longevity pathways. These findings will provide insight into fundamental mechanisms of aging and may facilitate the development of therapies to delay the onset and progression of age-related diseases in people.