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Cytochrome P450 Monooxygenase-Mediated Lipid Metabolism In Obesity And Colon Tumorigenesis, Weicang Wang Oct 2019

Cytochrome P450 Monooxygenase-Mediated Lipid Metabolism In Obesity And Colon Tumorigenesis, Weicang Wang

Doctoral Dissertations

Colon cancer is a major public health issue: it is expected to have 140,250 new cases and 50,630 deaths during 2018, making colon cancer the third most common type of cancer and the second leading cause of cancer death in the United States. Obesity is associated with enhanced colonic inflammation, which is a major risk factor of colorectal cancer. Currently, more than 35% of adults and nearly 17% of children are obese. Considering the obesity and colon cancer epidemic in the United States, there is an urgent need to identify novel therapeutic targets for obesity and colon cancer ...


Protein Detection And Structural Characterization By Mass Spectrometry Using Supramolecular Assemblies And Small Molecules, Bo Zhao Oct 2019

Protein Detection And Structural Characterization By Mass Spectrometry Using Supramolecular Assemblies And Small Molecules, Bo Zhao

Doctoral Dissertations

Mass spectrometry (MS) has played an increasingly prominent role in proteomics and structure biology because it shows superior capabilities in identification, quantification and structural characterization of proteins. To realize its full potential in protein analysis, significant progress has been made in developing innovative techniques and reagents that can couple to MS detection. This dissertation demonstrates the use of polymeric supramolecular assemblies for enhanced protein detection in complex biological mixtures by MS. An amphiphilic random co-polymer scaffold is developed to form functional supramolecular assemblies for protein/ peptide enrichment. The influences of charge density and functional group pKa on host-guest interactions ...


Improving Chinese Mothers’ Health Literacy: A Wechat Intervention, Qiong Chen Oct 2019

Improving Chinese Mothers’ Health Literacy: A Wechat Intervention, Qiong Chen

Doctoral Dissertations

The health literacy and eHealth literacy of women during the reproductive age is crucial, as it can affect their health and the health of their children. Promoting health literacy is essential to achieve mothers’ empowerment by improving access to and capacity of using health information effectively. However, functional, interactive, and critical health literacy and eHealth literacy have never been assessed among Chinese women.

The first study during this dissertation assessed functional, interactive, and critical health literacy and eHealth literacy among 421 of Chinese mothers with children under 3 years old. The results revealed overall less than optimal level of health ...


Multi-Level Barriers To Art Adherence Among Hiv-Infected Women In Rural Eswatini: A Mixed Methods Approach, Nozipho Becker Oct 2019

Multi-Level Barriers To Art Adherence Among Hiv-Infected Women In Rural Eswatini: A Mixed Methods Approach, Nozipho Becker

Doctoral Dissertations

Eswatini has the highest global prevalence of HIV despite universal access to free treatment. Lack of compliance continues to be a significant challenge for HIV care and management programs throughout the country. Studies investigating barriers to antiretroviral therapy (ART) adherence, particularly in women, who are the most vulnerable to HIV infections, are limited. The disproportionate impact of HIV on women can be attributed to multiple risk factors at the individual, household, and community/structural levels. Women living in rural settings are particularly at risk as a result of socio-cultural and environmental vulnerabilities such as oppressive traditional practices, intimate partner violence ...


Development And Cross-Validation Of A Cadence-Based Metabolic Equation For Walking, Christopher C. Moore Jul 2019

Mechanisms And Mitigation Of Skeletal Muscle Fatigue In Single Fibers From Older Adults, Aurora Foster Jul 2019

Mechanisms And Mitigation Of Skeletal Muscle Fatigue In Single Fibers From Older Adults, Aurora Foster

Masters Theses

Skeletal muscle fatigue is the contraction-induced decline in whole muscle force or power, and can be greater in older versus young adults. Fatigue primarily results from increased metabolism elevating phosphate (Pi) and hydrogen (H+), which alters myosin-actin interactions; however, which steps of the myosin-actin cross-bridge cycle are changed and their reversibility are unclear. PURPOSE: This study sought to: 1) Examine the effects of elevated Pi and H+ on molecular and cellular function, and 2) Test the ability of deoxyadenosine triphosphate (dATP), an alternative energy to adenosine triphosphate (ATP), to reverse the contractile changes induced with high Pi ...


Hybridized Polymeric Nano-Assemblies: Key Insights Into Addressing Mdr Infections, Ryan Landis Mar 2019

Hybridized Polymeric Nano-Assemblies: Key Insights Into Addressing Mdr Infections, Ryan Landis

Doctoral Dissertations

Multidrug-resistant (MDR) bacteria contribute to more than 700,000 annual deaths world-wide. Millions more suffer from limb amputations or face high healthcare treatment costs where prolonged and costly therapeutic regimens are used to counter MDR infections. While there is an international push to develop novel and more powerful antimicrobials to address the impending threat, one particularly interesting approach that has re-emerged are essential oils, phytochemical extracts derived from plant sources. While their antimicrobial activity demonstrates a promising avenue, their stability in aqueous media, limits their practical use in or on mammals. Inspired by the versatility of polymer nanotechnology and the ...


Evaluating The Toxicity And Formation Of Halobenzoquinones In Point-Of-Use Chlorinated Drinking Water, Stephanie Hung Oct 2018

Evaluating The Toxicity And Formation Of Halobenzoquinones In Point-Of-Use Chlorinated Drinking Water, Stephanie Hung

Masters Theses

Chlorine has effectively reduced the prevalence of waterborne diseases, however there are secondary consequences to this public health advancement. Disinfection byproducts (DBPs) are chemicals formed when chlorine reacts with natural organic matter (NOM) in water. A new class of DBPs, halobenzoquinones (HBQs), has recently been identified and data suggests it could be potentially carcinogenic and up to 1000 times more toxic than some regulated DBPs. So far, in vitro studies have assessed HBQ toxicity without taking into account its transformation in cell media into potentially less toxic compounds. This study evaluated the toxic effects of one HBQ, 2,6-DCBQ, and ...


Real-Time Dengue Forecasting In Thailand: A Comparison Of Penalized Regression Approaches Using Internet Search Data, Caroline Kusiak Oct 2018

Real-Time Dengue Forecasting In Thailand: A Comparison Of Penalized Regression Approaches Using Internet Search Data, Caroline Kusiak

Masters Theses

Dengue fever affects over 390 million people annually worldwide and is of particu- lar concern in Southeast Asia where it is one of the leading causes of hospitalization. Modeling trends in dengue occurrence can provide valuable information to Public Health officials, however many challenges arise depending on the data available. In Thailand, reporting of dengue cases is often delayed by more than 6 weeks, and a small fraction of cases may not be reported until over 11 months after they occurred. This study shows that incorporating data on Google Search trends can improve dis- ease predictions in settings with severely ...


Clpxp-Regulated Proteins Suppress Requirement For Reca In Dam Mutants Of Escherichia Coli K-12, Amie Savakis Oct 2018

Clpxp-Regulated Proteins Suppress Requirement For Reca In Dam Mutants Of Escherichia Coli K-12, Amie Savakis

Masters Theses

Double strand breaks (DSB) are a common source of DNA damage in both prokaryotes and eukaryotes. If they are not repaired or are repaired incorrectly, they can lead to cell death (bacteria) or cancer (humans). In Escherichia coli, repair of DSB are typically accomplished via homologous recombination and mediated by RecA. This repair pathway, among others, is associated with activation of the SOS response. DNA adenine methyltransferase (dam) mutants have an increased number of DSB and, therefore, are notorious for being RecA-dependent for viability. Here, we show that the synthetic lethality of Δdam/ΔrecA is suppressed when clpP is removed ...


Impact Of Bodyweight On Tissue-Specific Folate Status, Genome Wide And Gene-Specific Dna Methylation In Normal Breast Tissues From Premenopausal Women, Armina-Lyn Frederick Jul 2018

Impact Of Bodyweight On Tissue-Specific Folate Status, Genome Wide And Gene-Specific Dna Methylation In Normal Breast Tissues From Premenopausal Women, Armina-Lyn Frederick

Masters Theses

Obesity has reached an epidemic level in the United States. A number of epidemiological studies have established obesity as a critical risk factor for postmenopausal breast cancer (post-BC), whereas a reverse association holds prior to menopause. A significant scientific gap exists in understanding the mechanism(s) underpinning this epidemiological phenomenon, particularly the reverse association between obesity and premenopausal breast cancer (pre-BC). This study aimed to understand how folate metabolism and DNA methylation informs the association between obesity and pre-BC. Fifty normal breast tissue samples were collected from premenopausal women who underwent reduction mammoplasty. We developed and measured the breast tissue ...


Sleep, And Its Relation To Non-Motor Deficits In Patients With Cerebellar Ataxia, Akshata Sonni Mar 2018

Sleep, And Its Relation To Non-Motor Deficits In Patients With Cerebellar Ataxia, Akshata Sonni

Doctoral Dissertations

The cerebellum is a highly connected structure, and its involvement in sleep – which is a dynamic process that is modulated by a complex set of neural systems – can come about through a number of neural pathways. We conducted two studies aimed at furthering our understanding of cerebellar involvement in sleep behavior and physiology, as well as measuring the impact of poor sleep on mood and cognition in patients with cerebellar degeneration. First, by means of an online battery including measures of sleep and neuropsychiatric function, we collected data from 176 patients with cerebellar ataxia. We found strong evidence of poor ...


Robust Biosensors For Healthcare Applications: From High-Content Screening To Point-Of-Care Testing, Ngoc D. B. Le Nov 2017

Robust Biosensors For Healthcare Applications: From High-Content Screening To Point-Of-Care Testing, Ngoc D. B. Le

Doctoral Dissertations

Efficient detection of proteins, mammalian cells, microorganisms and other biological systems in complex mixture is essential in disease diagnosis and environmental health. Therefore, technological platforms that provide sensors of high sensitivity, selectivity and stability are greatly desired. Recently, the ‘chemical-nose’ sensing approach has proved to be an effective strategy for profiling bio-relevant targets in complex mixtures. Detecting analytes in complex mixture is a challenge that conventional specificity-based sensors are still trying to solve due to the requirement of prior knowledge of the analyte, which is unknown in many cases. This thesis focuses on how to develop simple and robust ...


Effects Of Malformed Or Absent Valves To Lymphatic Fluid Transport And Lymphedema In Vivo In Mice, Akshay S. Pujari Oct 2017

Effects Of Malformed Or Absent Valves To Lymphatic Fluid Transport And Lymphedema In Vivo In Mice, Akshay S. Pujari

Masters Theses

Lymph is primarily composed of fluid and proteins from the blood circulatory system that drain into the space surrounding cells, interstitial space. From the interstitial space, the fluid enters and circulates in the lymphatic system until it is delivered into the venous system. In contrast to the blood circulatory system, the lymphatic system lacks a central pumping organ dictating the predominant driving pressure and velocity of lymph. Transport of lymph via capillaries, pre-collecting and collecting lymphatic vessels relies on the synergy between pressure gradients, local tissue motion, valves and lymphatic vessel contractility. The direction of lymph transport is regulated by ...


Extractable And Non-Extractable Polyphenols From Apples: Potential Anti-Inflammatory Agents, Makenzi Gennette Oct 2017

Extractable And Non-Extractable Polyphenols From Apples: Potential Anti-Inflammatory Agents, Makenzi Gennette

Masters Theses

With diet being such a huge factor in the development of diseases, emerging research has supported that apple consumption is a promising candidate for disease prevention due to the high phenolic content it possesses. These polyphenols can be found in two forms: extractable polyphenols (EP) and non-extractable polyphenols (NEP). Polyphenols have been shown to have strong anti-inflammatory and anti-oxidant properties, but up until this point, most researchers focus on EP fractions, while NEP are neglected. After the EP extraction using acetone and acetic acid (99:1) from the Apple Boost powder, three additional extraction methods were conducted on the remaining ...


Total Organic Iodine Quantification And Occurrence In Drinking Water, And Toxicity Assessment Of Iodinated Disinfection By-Products, Rassil El Sayess Mar 2017

Total Organic Iodine Quantification And Occurrence In Drinking Water, And Toxicity Assessment Of Iodinated Disinfection By-Products, Rassil El Sayess

Doctoral Dissertations

The focus of this work has been placed is on iodinated DBPs (I-DBPs), measured using total organic iodine (TOI), a surrogate measure of iodinated organics. This is due to the growing toxicity literature that places I-DBPs among the most toxic of all DBPs. To measure TOI in water, a new method was developed. This method combines adsorption, combustion, and trapping of combustion products, with an offline inductively coupled plasma/mass spectrometer (ICP-MS) for iodide detection. Three factors were varied across two levels each in order to optimize the method. The chosen method used a sample pH of less than 1 ...


Alternative Methods For The Treatment Of Chemo-Resistant Cancers, Kaitlyn Wong Jul 2016

Alternative Methods For The Treatment Of Chemo-Resistant Cancers, Kaitlyn Wong

Doctoral Dissertations

Great strides have been made in cancer therapy in the past century, yet it remains one of the leading causes of death in the United States today. This work aimed to shed light on novel methods to treat a variety of aggressive and often chemo-resistant cancers both in vitro and in vivo.

The first aim of this work was to evaluate the therapeutic efficacy of poly(methacryloyloxyethyl phosphorylcholine) (polyMPC) prodrugs compared to standard chemotherapeutic agents. Conjugation of polyMPC to drugs such as doxorubicin (Dox) can result in its improved solubility, prolonged half-life and therapeutic efficacy. PolyMPC and polyMPC-Dox (at a ...


Macronutrients And The Risk Of Premenstrual Syndrome, Serena C. Houghton Jul 2016

Macronutrients And The Risk Of Premenstrual Syndrome, Serena C. Houghton

Doctoral Dissertations

Premenstrual syndrome (PMS) affects 8-20% of reproductive-aged women, impacting work, family, and social interactions. Limitations in available PMS treatments, including side effects and limited medication efficacy, indicate the need for improved prevention. Modifiable risk factors for prevention of PMS include dietary factors. Several micronutrients have been identified as risk factors, but there has been little evaluation of macronutrients. Thus, the research aim was to examine prospectively whether macronutrient consumption was associated with PMS development among a subset of women enrolled in the Nurses’ Health Study II cohort.

Chapter 1 evaluates the association of fat intake and PMS risk. Among 3 ...


Feasibility Of A Combined Physical Activity And Sleep Education Intervention For Girls Living In A Low Socioeconomic Status Urban Community, Cory J. Greever Jul 2016

Feasibility Of A Combined Physical Activity And Sleep Education Intervention For Girls Living In A Low Socioeconomic Status Urban Community, Cory J. Greever

Doctoral Dissertations

Declining sleep duration and poor sleep quality is a public health epidemic disproportionately effecting elementary age girls living in low socioeconomic status urban communities. The Girls Dancing and Sleeping for Health (Girls DASH) program was designed to test the feasibility, acceptability, and efficacy of a combined physical activity and sleep education program in this population. In combination with baseline data from a previously conducted intervention in elementary age urban girls living in a low SES community in Springfield, MA, results from a cross-sectional analysis indicated that greater screen time was positively associated with parental reports of child’s sleep quality ...


Evaluating A Novel Photochemical Tool For Labeling And Tracking Live, Endogenous Calcium-Permeable Ampars, Rosamund Elizabeth Combs-Bachmann Jul 2016

Evaluating A Novel Photochemical Tool For Labeling And Tracking Live, Endogenous Calcium-Permeable Ampars, Rosamund Elizabeth Combs-Bachmann

Masters Theses

The purpose of this research is to advance development of a photochemical tool designed to probe the role of ionotropic glutamate receptor signaling in neurodegenerative processes, and to delve more deeply into the biological processes underlying the role of these receptors in signaling and memory formation. This ligand-targeted nanoprobe was designed and developed in our lab to label endogenous calcium-permeable AMPARs (CP-AMPARs) in live cells with minimal disruption to native receptor activity. Nanoprobe is designed to use naphthyl acetyl spermine (NASPM) as a photocleavable ligand to target and covalently label native CP-AMPARs with a non-perturbing, fluorescent marker that then allows ...


The Human Intruder Test: An Anxiety Assessment In Rhesus Macaques (Macaca Mulatta), Emily J. Peterson Nov 2015

The Human Intruder Test: An Anxiety Assessment In Rhesus Macaques (Macaca Mulatta), Emily J. Peterson

Masters Theses

The human intruder test (HIT) is a noninvasive tool widely used for assessing anxiety in rhesus macaques (Macaca mulatta). This thesis explores the HIT procedure and applies it to a population of monkeys with a self-injurious behavioral pathology. Individual variation on this test can be used to assess anxiety and temperament. The first experiment of this thesis applied two different procedures of the HIT to 17 monkeys at UMass. Monkeys displayed little response to the intruder, and no significant differences were detected for the two procedures. To determine whether these responses were unique to the UMass monkeys, their behavior was ...


Engineering Novel Detection And Treatment Strategies For Bacterial Therapy Of Cancer, Jan T. Panteli Aug 2015

Engineering Novel Detection And Treatment Strategies For Bacterial Therapy Of Cancer, Jan T. Panteli

Doctoral Dissertations

Finding and treating cancer is difficult due to limited sensitivity and specificity of current detection and treatment strategies. Many chemotherapeutic drugs are small molecules that are limited by diffusion, making it difficult to reach cancer sites requiring high doses that lead to systemic toxicity and off-target effects. Tomographic detection techniques, like PET, MRI and CT, are good at identifying macroscopic lesions in the body but are limited in their ability to detect microscopic lesions. Biomarker detection strategies are extremely sensitive and able to identify ng/ml concentrations of protein, but are poor at discriminating between healthy and disease state levels ...


Impact Of A Preschool Workplace Intervention On Classroom Teachers’ Physical Activity Levels And Quality Of Life, Ogechi O. Nwaokelemeh Aug 2015

Impact Of A Preschool Workplace Intervention On Classroom Teachers’ Physical Activity Levels And Quality Of Life, Ogechi O. Nwaokelemeh

Doctoral Dissertations

PURPOSE: To examine the feasibility and efficacy of a three-month, cluster-randomized workplace PA intervention on PA levels and wellbeing measures in preschool classroom teachers. METHODS: Ten preschool centers were randomized into two groups; short-bouts of structured PA (SBS-PA, n=5) or traditional unstructured PA (TRAD-PA, n=5). The SBS-PA intervention consisted of 10-minute structured, teacher-led PA routines implemented within the classroom setting, followed by 20 minutes of unstructured playtime. The TRAD-PA intervention consisted of supervised 30-minute of unstructured free playtime. Both interventions were implemented during the morning and afternoon designated gross-motor playtime for 30 min/session, five days/week for ...


Mimicking The Arterial Microenvironment With Peg-Pc To Investigate The Roles Of Physicochemical Stimuli In Smc Phenotype And Behavior, William G. Herrick Aug 2015

Mimicking The Arterial Microenvironment With Peg-Pc To Investigate The Roles Of Physicochemical Stimuli In Smc Phenotype And Behavior, William G. Herrick

Doctoral Dissertations

The goal of this dissertation was to parse the roles of physical, mechanical and chemical cues in the phenotype plasticity of smooth muscle cells (SMCs) in atherosclerosis. We first developed and characterized a novel synthetic hydrogel with desirable traits for studying mechanotransduction in vitro. This hydrogel, PEG-PC, is a co-polymer of poly(ethylene glycol) and phosphorylcholine with an incredible range of Young’s moduli (~1 kPa - 9 MPa) that enables reproduction of nearly any tissue stiffness, exceptional optical and anti-fouling properties, and support for covalent attachment of extracellular matrix (ECM) proteins. To our knowledge, this combination of mechanical range, low ...


The Effects Of Ovarian Hormones And Exercise On Gene Markers Of Cardiac Dysfunction, Anisha S. Patel Jul 2015

The Effects Of Ovarian Hormones And Exercise On Gene Markers Of Cardiac Dysfunction, Anisha S. Patel

Masters Theses

Heart disease is the leading cause of death in women in the United States. Premenopausal women appear to have better cardiac function and lower risk of heart disease compared to male postmenopausal female counterparts. Ovarian hormone loss influences blood pressure homeostasis and causes systemic inflammation, which may result in chronic stress on the heart. Two key physiological changes in cardiac dysfunction are reemergence of the fetal gene pattern and myocardial remodeling. Physical activity has been linked to improved cardiac function. The purpose of this study was to investigate the effects of ovariectomy on early markers of cardiac dysfunction and fibrosis ...


Thermo-Responsive Poly(N-Isopropylacrylamide) And Its Critical Solution Temperature Type Behavior In Presence Of Hydrophilic Ionic Liquids, Purnendu K. Nayak Mar 2015

Thermo-Responsive Poly(N-Isopropylacrylamide) And Its Critical Solution Temperature Type Behavior In Presence Of Hydrophilic Ionic Liquids, Purnendu K. Nayak

Masters Theses

Thermo-responsive homopolymer poly(N-isopropylacrylamide), is a widely studied and used polymer. Our recent observations on thermal behavior of aqueous solutions of this polymer requires a short overview of existing results in order to understand the formation of different phases, both stable and unstable with the addition of hydrophilic Ionic liquids (ILs) 1-Butyl-3-methylimidazolium tetrafluoroborate ([BMIM][BF4]), 1-Butyl-3-methylimidazolium acetate ([BMIM][OAc]) and 1-Butyl-3-methylimidazolium thiocyanate ([BMIM][SCN]) to the system. PNIPAM is soluble in cold water due to its inter- and intramolecular hydrogen bonding but phase separates upon heating at T > 32 , which is its lower critical solution temperature (LCST). PNIPAM exists ...


The Role Of The Extracellular Matrix In Mediating Muscle Soreness, Karen L. Riska Mar 2015

The Role Of The Extracellular Matrix In Mediating Muscle Soreness, Karen L. Riska

Doctoral Dissertations

The goal of this dissertation was to examine the role of the extracellular matrix (ECM) in muscle soreness. Study 1 examined how the disruption of the ECM from a minor surgery (e.g. muscle biopsy) affected muscle soreness. Study 1 showed that soreness levels increased at 24 h post-biopsy compared to baseline and resolved within 96 h. However, the level of muscle soreness at 24 h post-biopsy (20 mm) was lower than that reported for performing a strenuous and naïve exercise (40–80 mm). These results will help review boards at institutions where muscle biopsies are performed understand how a ...


A Novel Population Of Natural Killer Cells Plays A Critical Role In The Depletion Of Splenic B2 B Cells During Experimental African Trypanosomiasis, Deborah Frenkel Mar 2015

A Novel Population Of Natural Killer Cells Plays A Critical Role In The Depletion Of Splenic B2 B Cells During Experimental African Trypanosomiasis, Deborah Frenkel

Doctoral Dissertations

Loss of humoral immune competence in T. brucei-infected mice is associated with the apoptotic depletion of splenic transitional, marginal zone and follicular B cells as well as a depletion of CD8+ T cells. This occurs rapidly after infection and impairs responses to vaccine antigens in addition to responses to newly arising VSG antigenic variants, leading to uncontrolled parasite growth and death of the infected mice. Infection-induced B2 B cell and CD8+ T cell loss requires the presence of a novel population of natural killer (NK) cells and is mediated by a perforin-dependent process consistent with perforin- and granzyme-mediated cytotoxicity ...


Elucidating Cellular Signaling Pathways That Contribute To The Immunopathogenesis Of Aplastic Anemia, Christina M. Kuksin Nov 2014

Elucidating Cellular Signaling Pathways That Contribute To The Immunopathogenesis Of Aplastic Anemia, Christina M. Kuksin

Doctoral Dissertations

Aplastic Anemia is a rare immune mediated bone marrow failure disease that is mediated by autoreactive T cells that cause destruction to the stem, progenitor, and stromal cells in the bone marrow. Because little is known about the etiology of the disease, our lab has developed a major MHC mismatch GVHD model to examine cellular pathways in autoreactive T cells that contribute to disease. We examined three pathways that are important in T cell activation and differentiation and asked if they were important in the development of Aplastic Anemia. First, we were able to show that expression of active PKCθ ...


Dried Whole Plant Artemisia Annua As A Novel Antimalarial Therapy, Mostafa A. Elfawal Nov 2014

Dried Whole Plant Artemisia Annua As A Novel Antimalarial Therapy, Mostafa A. Elfawal

Doctoral Dissertations

Dried Whole plant Artemisia annua as a novel antimalarial therapy

September 2014

Mostafa Ahmed Elfawal

Ph.D., UNIVERSITY OF MASSACHUSETTS AMHERST

Directed by: Professor Stephen M. Rich

Malaria is one of the worst vector-borne parasitic diseases in the developing world. The World Health Organization (WHO) estimated that 215 million cases of malaria occurred, with >655,000 deaths; half the world’s population is at risk of contracting the disease. Drugs are primary weapons for reducing malaria in human populations. Successful drugs are highly efficacious and inexpensive to manufacture synthetically. However, emergence of resistant parasites has repeatedly curtailed the lifespan of ...