Ataxia-telangiectasia mutated kinase (ATM), a serine/threonine kinase primarily located in the nucleus, is typically activated in response to DNA damage. Individuals with mutations in ATM gene develop a disease called Ataxia telangiectasia (AT). These individuals are more susceptible to ischemic heart disease and metabolic disorder. Our lab has previously shown that ATM plays a critical role in β-adrenergic receptor (β-AR) - and myocardial infarction (MI)-stimulated myocyte apoptosis and cardiac remodeling. This study tested the hypothesis that ATM plays a critical role in cardiac autophagy and glucose metabolism following MI and ischemia, respectively. Early during MI (4 hours after its onset) and 4 hours post-treatment with ATM inhibitor KU-55933, ATM deficiency resulted in autophagic impairment in the heart and in cardiac fibroblasts, respectively. Such autophagic changes in the heart and in cardiac fibroblasts associated with the activation of GSK-3β and mTOR, and inactivation of Akt and AMPK. ATM deficiency also augmented autophagy in the infarct region of the heart 28 days post-MI as well as in cardiac fibroblasts treated with ATM inhibitor KU-55933 for 24 hours. Autophagic changes in the infarct region during ATM deficiency associated with enhanced Akt, Erk1/2, and mTOR activation. Additionally, the lack of ATM accelerated glycolysis and gluconeogenesis and augmented TCA cycle metabolism under non-ischemic conditions. Following a 20 minute global ischemic period, the glycolytic pathway, not the gluconeogenic pathway, was down-regulated during ATM deficiency which was found to be associated with alterations in TCA cycle metabolism. Such metabolic rearrangements associated with changes in the phosphorylation of Akt, GSK-3β, and AMPK alongside alterations in Glut4 protein expression. Thus, ATM deficiency impairs autophagy early after the onset of MI and in cardiac fibroblasts treated with ATM inhibitor KU-55933 for 4 hours. In contrast, ATM deficiency appears to augment autophagy late post-MI in the infarct region of the heart and in cardiac fibroblasts treated with ATM inhibitor KU-55933 for 24 hours. Lack of ATM alters glucose and TCA cycle metabolism with and without ischemia. Such findings implicate ATM as a key player in autophagic changes in the heart in response to MI as well as in glucose metabolism under non-ischemic and ischemic conditions.

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No description available.

In 1882, Sydney Ringer, a professor of medicine at University College in London, experimented with the frog ventricle to better understand how each constituent of blood influences contraction. The ultimate goal was to create an artificial circulating fluid to use for the perfusion of isolated organs, in this case, a frog heart. Today, Ringer’s solution is still used in research for physiological studies requiring the survival and maintenance of specimens outside of their host bodies. One such example is the use of medicinal leech ganglia for electrophysiological measurements. In this thesis, I am comparing two Ringer’s solutions, original versus added glucose, and their impact on the longevity of the ganglia. By stimulating cells in the dissected ganglia submerged in Ringer’s solution with a micropipette, action potential responses can be recorded and used to compare longevity of the cells in each solution. By providing the dissected ganglia with an additional source of fuel, I hypothesized that cells in the glucose-enriched Ringer’s solution would live longer, and thus provide action potentials longer, than cells in regular Ringer’s solution with a minimum increase in longevity of thirty minutes. Data analysis showed that glucose Ringer’s solution did not keep the cells alive longer than regular Ringer’s solution when the difference of means was set to 30 minutes. However, data did show a significant difference in the average longevity of the Retzius cell in glucose Ringer’s solution versus regular Ringer’s solution when the difference of means was set to zero.

A pig model of childhood obesity was used to study the effects of dietary energy on body adiposity, and blood parameters associated with impaired glucose clearance. Prepubertal female pigs weaned at 21 d of age were fed control (CON), refined sugar (SUG), fat (FAT), and sugar-fat (SUGFAT) diets in a completely randomized arrangement for 16 wk. Calories from fat were 8.9% for CON, 5.6% for SUG, 35.5% for FAT and 32.3% for SUGFAT. Calories from sugar were 36.0% for SUG and 30.7% for SUGFAT. Adding fat, sugar or both to diets increased (P < 0.003) calorie intake. Percentage body fat was higher (P < 0.0001) in all treatments compared to CON, and in SUGFAT and FAT compared to SUG. Ultrasound back fat depth was positively correlated (r2 = 0.909; P < 0.001) with percentage body fat and negatively (r = 0.912; P-value ) with percentage body protein. Area under the curve (AUC) in response to oral glucose tolerance at 14 wk was higher (P < 0.03) in FAT (+14.6%) and SUGFAT (+25.5%) pigs compared to CON. Glucose AUC from sugar-fed pigs was not different (P = 0.2) from fat alone-fed pigs. Adding sugar, fat, or their combination to diets increased (P < 0.008) blood glucose and decreased (P < 0.0009) plasma insulin AUC. These data show that inclusion of fat and refined sugar in pig diets increases body adiposity and impairs glucose homeostasis and suggests that the composition of calories consumed may have different effects than simply consumption of excess of calories. / Master of Science

<p> Poly (dimethyl siloxane) (PDMS) has been widely used as a biomaterial in ophthalmic and other applications due to its good compatibility, high mechanical strength and excellent oxygen permeability and transparency. For use as an artificial cornea, contact lens and in other applications, modifications are necessary to improve glucose permeability and wettability for cell and tear protein and mucin interactions through modification with hydrophilic functional groups or polymers. Poly (N-isopropyl acrylamide) (PNIPAAM) is a biocompatible and hydrophilic polymer that has been extensively studied in controlled drug release applications due to its lower critical solution temperature (LCST) phenomenon. In this study, a composite interpenetrating polymer network (IPN) of PDMS and PNIPAAM was formed to generate material with reasonable oxygen and glucose permeability as well as improved wettability and mechanical properties compared to the PDMS and PNIPAAM homopolymers.</p> <p> Semi-IPNs, with low water uptake and mechanical strength, were found not to be suitable as biomaterials. Vinyl terminated PDMS/PNIPAAM IPNs had reasonable water uptake and excellent tensile stress and strain, but low glucose permeability (< 10^-10 cm^2/s). Hydroxyl terminated PDMS/PNIPAAM IPNs (PDMS-OH IPN) were successfully synthesized with reasonable mechanical properties and significantly higher glucose permeability (~10^-7 cm^2/s). Curing the PDMS-OH film with solvent was found to improve glucose transport.</p> <p> The presence of PNIPAAM in the composite networks was confirmed by FT-IR and Differential Scanning Calorimetry (DSC). Transmission Electron Microscopy (TEM) images verified the structure of interpenetrating networks. Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR) and X-ray Photoelectron Spectroscopy (XPS) suggested that PNIPAAM was also present on the surface and this translated to increased roughness compared with the PDMS control as determined by AFM. The LCST phenomena still remained in the IPN, although the change was not as abrupt as with pure PNIPAAM. These results suggest that the copolymer may be useful as an ophthalmic biomaterial and for controlled drug release applications.</p> / Thesis / Master of Applied Science (MASc)

A thesis submitted to the School of Graduate Studies in partial fulfillment of the requirements for the degree Master of Science. / Sprint interval training (SIT) is a time-efficient strategy to improve cardiorespiratory fitness; however, most protocols have been studied in a laboratory setting and require specialized equipment. We investigated the efficacy of brief intense stair climbing as a practical model of SIT to improve cardiometabolic health, with a key measure being cardiorespiratory fitness as indicated by peak oxygen uptake (VO2peak). Two separate studies, each consisting of an acute and chronic phase, were conducted in a total of 31 sedentary women (age=24±10 y; BMI=23±4 kg•m-2). The acute phase of Study 1 established that the heart rate (HR), blood lactate concentration (BLa), and rating of perceived exertion (RPE) responses were similar when participants (n=8) performed a SIT protocol that involved 3x20-s “all-out” efforts of either continuous stair climbing or cycling, interspersed with 2 min of recovery. The chronic phase demonstrated that when participants (n=12) performed the 3x20-s stair climbing protocol 3 d•wk-1 for 6 wk, absolute and relative VO2peak increased by 12%, or ~1 metabolic equivalent (1.80±0.25 to 2.02±0.27 L•min-1, p<0.001), as there were no changes in body mass (p=0.35), fat free mass (FFM; p=0.09) or % body fat (p=0.42). There were also no changes in resting systolic and diastolic blood pressure (BP; p=0.82 and p=0.97, respectively), resting HR (p=0.62), and fasting insulin sensitivity (p=0.52). The acute phase of Study 2 established that the HR and RPE responses were similar when participants (n=11) performed three different stair climbing protocols. The protocols investigated include the 3x20-s continuous ascent model used in Study 1 (protocol 1), as well as 3x60-s bouts of ascending and descending either one or two flights of stairs, with 60-s of recovery (protocol 2 and 3, respectively). The chronic phase demonstrated that when the same group of subjects performed the 3x60-s 1-flight protocol 3 d•wk-1 for 6 wk, absolute and relative VO2peak increased by 8 and 7%, respectively (1.79±0.36 to 1.93±0.39 L•min-1, p=0.001; 31.2±4.6 to 33.3±5.3 mL•kg-1•min-1; p=0.01). Despite no changes in % body fat (p=0.10), there was an increase 3% increase in FFM (p<0.001). There was no change in systolic (p=0.50) and diastolic BP (p=1.00), but resting HR improved by 8% after training (p=0.03). The change in insulin sensitivity derived from an OGTT was 7.1±11 mg I2•mmol-1•mIU-1•min-1 (p=0.056). These findings demonstrate that brief intense stair climbing is a practical, time-efficient strategy to improve cardiorespiratory fitness in previously untrained women. / Thesis / Master of Science (MSc) / Sprint interval training (SIT), involving brief bouts of very intense exercise separated by short periods of recovery, is a time-efficient alternative to traditional endurance training for improving fitness. This has largely been established in laboratory settings using specialized equipment, which is impractical for many individuals. This project examined whether brief intense stair climbing was a practical model of SIT to elicit adaptations previously shown with cycling protocols. Subjects performed either three 20-s ascents interspersed with 2 min recovery periods, or three 60-s bouts of ascending and descending one or two flights of stairs, with 60-s recovery periods. Both protocols were 10 min in duration including warm-up and cool-down, and subjects trained three days per week for six weeks. The main finding was that stair climbing is a practical, time-efficient model to improve fitness in previously sedentary individuals.

Overweight and obesity has increased in the U.S. Unfortunately, this increases the risk of metabolic dysfunction, such as diabetes. Meal timing has recently shown promise as viable options to minimize the risk of overweight or obesity and metabolic dysfunction. The primary purpose of this study, therefore, was to investigate the effect of timing of energy intake on blood glucose in females with overweight or class one obesity (BMI=27-35 kg/m2). This study utilized crossover design with two conditions counterbalanced in random order. Participants completed two 2-week 1500kcal dietary interventions: 1) a big breakfast diet or BB (50% of energy intake between 7-9am, 35% of energy between 12-2pm, and ~15% of energy between 5-7pm), and 2) a big dinner diet or BD (~15% of energy intake between 7-9am, of energy between 12-2pm, and ~50% of energy between 5-7pm). There was a 2-week wash-out period between each condition. Meals were provided to participants. The primary outcome was differences in glucose over the course of each 2-week period. Glucose was determined using a continuous glucose monitor and analyzed for area under the curve (AUC) using the trapezoidal method. Other exploratory outcomes were assessed, including: fasting glucose, height, weight, body composition using dual-energy x-ray absorptiometry, physical activity and sleep using accelerometry, waist circumference, hunger using visual analog scales, sleep quality using the Pittsburg Sleep Quality Index, depression and anxiety using the Depression Anxiety and Stress Scale, eating behaviors using the Dutch Eating Behavior Questionnaire, resting heart rate using an oximeter, blood pressure using an aneroid cuff and sphygmomanometer and temperature using a temporal scan thermometer. This study was designed for proof of concept. The primary analysis showed there was no significant difference in total AUC glucose between the BB and BD conditions (F=0.14; p=0.7137). A secondary analysis indicated a significant interaction between dietary condition across the 14-day intervention (F=98.23; p<0.001) and condition x hour interaction for glucose AUC (F=32.62; p<0.0001). As expected, the BB diet had higher AUC glucose from 7am-12pm (F=15.48; p=0.001) and the BD diet has higher AUC glucose from 5pm-10pm (F=11.63; p=0.0031). Both conditions were effective to decrease weight, waist circumference, BMI, and fat free mass (p<0.05); however, the condition x time interaction was significantly more reduced for fat mass during the BB than the BD condition (F=4.37; p=0.0411). This study did not reveal a difference in total AUC glucose between the BB and BD. It was shown that the BB saw a greater rate of fat mass loss then the BD. This indicates that meal timing can provide addition benefits to the outcomes caused by a caloric deficit.

The pathomechanism of migraine attacks is not understood well, however, is currently believed to be a brain disorder. Migraine is a multifactorial disorder that needs to be investigated in multiple research areas to shed light into its mechanism and find ways to treat it effectively. Migraine episodes come from one or many things at the same time, and such vary from person to person. However, sufficient evidence in recent studies show there is a strong relationship between genetics with patients having specific genes that may be responsible for the disease or a genetic sequence passed down through generations in families, especially in the case of migraine with aura. Several genes including P/Q type calcium channel gene mutations, Na+ /K+ pump ATP1A2 malfunctions and Na+ gated voltage channel Nav 1.1 have been implicated with familial hemiplegic migraines with aura (Samsam 2012). Additionally, insulin resistance has been shown to be a very prevalent factor in migraine patients, but more research has yet to be done to support such claim. Also, many studies suggest the possibility of migraine attacks occurring due to a lack of energy in the patient´s brain. Meaning that exhaustion of the brain can also lead to a migraine episode. In this thesis, we investigated the genetic causes of migraine and reviewed the recent genes implicated in the pathomechanism of migraines. Further, we explored several articles that investigated the metabolic changes that occur in the brain during a migraine attack and looked for reasons behind such changes.

The electrochemical enzymatic electrodes dominate the world market for blood glucose monitoring devices for controlling, as well as reducing the detrimental effects of diabetes. However, the enzymatic electrodes exhibit constraints restricting their reliance on the enzyme’s activity which can be influenced by the external, and the environmental factors such as temperature, pH, and humidity etc. However, the greater thickness of the enzyme layer hinders the performance of the glucose biosensors resulting in signal dampening or loss. In addition, the selectivity of the electrodes is affected by the interferents present in blood. Moreover, the invasive nature of the electrodes is a major problem considering the patient’s perspective. In contrast, recent research activities demonstrated that the electrochemical non-enzymatic electrodes possess huge potential for inexpensive and highly sensitive glucose monitoring devices, yet these electrodes are invasive in nature. Therefore, the purpose of this research was to fabricate electrochemical non-enzymatic non-invasive electrodes for sweat glucose monitoring devices. A very simple low-cost fabrication technique has been shown to make the facile, flexible, and inexpensive electrodes to detect sugar in sweat bio-analyte for a non-invasive glucose monitoring system using the native stable Cu oxides (CuNOx), Cu2O, layers grown on 35 µm thin Cu foils keeping under ambient conditions (25℃- and 760-mm Hg) for more than 2 years so that the oxide layers are full-grown, and fully stable. Moreover, the foils also annealed at various temperatures such as 160, 230, and 280℃ with new temperature profile for reducing the required time of growing stable oxides and producing oxides with larger crystallized structures with higher surface – to – volume ratio. The X-ray photoelectron spectroscopy (XPS) and high-resolution transmission electron microscopy (HRTEM) results supported that at 280℃ annealing temperature the surface, mostly, transformed into highly electrocatalytic CuO with larger grain sizes, crystallized structures, and the uniform layer of ~ 140 nm. The electrochemical characterization, and sensing performance of the electrodes have been done by cyclic voltammetry (CV), one of the excellent and well accepted electrochemical methods, with the 3 – electrode configuration of the potentiostat. The CuNOx sensors of having ~10 nm layer of stable Cu2O exhibited a sensitivity of 603.42 μA mM−1 cm−2, a linear range beyond the desired limit of 7.00 mM with excellent linearity (R2 = 0.9983) and a low limit of detection of 94.21 μM. In contrast, the new annealing profile has. the CuNOx sensors annealed at 280 ºC using new temperature profile provided twin calibration curves of linear ranges of 0.05 – 1.00 mM and 1.00 – 7.00 mM, that applicable for sweat and blood glucose sensing, respectively, and exhibited a sensitivity of 1795 μA mM−1 cm−2, a linear range up to the desired limit of 1.00 mM for sweat glucose sensing with excellent linearity (R2 = 0.9844), and a lower limit of detection of 135.39 μM. In addition, it has been shown that the peak electro-oxidation current of glucose sensing is linearly related with the squire root of the annealing temperature, √T. This can help to figure out the required applied annealing temperature for getting desired peak electro-oxidation current of glucose in a human health monitoring system. / Dissertation / Doctor of Philosophy (PhD)