Low back pain is one of the major health concerns in the US. It affects up to 80% of the population at some time during their lives. It not only causes discomfort to patients and affects their physical ability but also has a huge economic impact on society. Although the cause of low back pain is still poorly understood, it is implicated that degeneration of the intervertebral disc is the primary factor. Currently, researchers are trying to use tissue engineering approaches to develop new treatments capable of removing the degenerated disk and replacing it with a biological substitute. However, to create such a biological substitute, we need to first understand the structure-function relationship of the tissue. Only when we understand the function of the tissue, can we begin creating biological substitutes. While culturing a biological substitute, we also need methods to determine how the substitute responds to its environment. At present, there are many different types of bioreactors developed for cartilaginous tissues. However, there is a lack of a system that can detect the chemical, electrical and mechanical response noninvasively with control feedback in real-time. It is hard to provide the optimal culture environment to the substitute without knowing its response in real-time. The objective of this dissertation is to develop new methods to investigate the transport property, oxygen consumption rate and mechano-electrochemical and mechanical properties of the tissue. Because cells are responsible for the tissue health, it is necessary to understand how they can obtain nutrients under different environments, e.g. under different loading condition. In addition, with the use of a bioreactor with the capability of detecting the real-time response combined with a feedback control system, we can provide the most favorable conditions for tissue or biological substitutes to grow. The new measurement methods developed in this dissertation can contribute to further understanding the function of the tissue. The methods outlined in this dissertation can also provide new tools for future tissue engineering applications. Moreover, the findings in this dissertation can provide information for developing a more comprehensive theoretical model to elucidate the etiology of disc degeneration.

The effect of temperature on predator and herbivore consumption is an important factor for predicting the effects of climate warming on ecosystems. The Metabolic Theory of Ecology (MTE) describes the temperature dependence of biological and ecological rates and states that metabolism is the fundamental biological mechanism that governs most observed patterns in ecology. This statement has been criticized empirically for a number of organismal traits and systematic deviations have been found. Here, a meta-analysis is performed on published temperature responses of ectotherm consumption. The mean effect of temperature on consumption was higher than the mean value predicted by proponents of the MTE and was highly variable. Some of this variation is explained by habitat type, where the consumption rates of marine organisms displayed stronger temperature dependence than for terrestrial and freshwater organisms. The frequency distribution of temperature dependencies is right skewed for consumption. Here, this skewness is explained by a methodological artefact as values close to “no effect” are more unlikely to be sampled than others when fitting the Arrhenius equation. In conclusion, the assumptions of the MTE do not hold for rates of consumption and marine organisms display a stronger temperature dependence compared to terrestrial and freshwater organisms.

Mitochondrial uncouplers have the capacity of passively shuttling protons from the mitochondrial intermembrane space to the mitochondrial matrix, independent of ATP synthase. This results in the disruption of oxidative phosphorylation and increased rate of metabolism as a counter action from the mitochondria. Therefore, small molecule mitochondrial uncouplers have potential for the treatment of obesity, diabetes, non-alcoholic fatty liver disease (NAFLD), neurodegenerative disorders, amongst others. A one-pot method for the synthesis of 1H-imidazo[4,5-b]pyrazines from [1,2,5]oxadiazolo[3,4-b]pyrazines is herein disclosed. In the presence of Fe, Yb(OTf)3, and the desired electrophile partner, in situ reduction of the oxadiazole fragment followed by cyclization afforded imidazolopyrazines in moderate to good yields. The selection of different orthoesters as electrophiles also allowed functionalization on the 2-position of the imidazole ring. This new method was used to synthesize 1H-imidazo[4,5-b]pyrazines to perform structure-activity relationship studies. Thus, a library of 75 compounds was synthesized and characterized for mitochondrial uncoupling activity. The biological activity of the compounds was demonstrated in oxygen consumption rate assays affording potent mitochondrial uncouplers. The method was further applied to the synthesis of 5-alkoxy-2-(trifluoromethyl)-1H-imidazo[4,5-b]pyrazin-6-amines, with over 50 derivatives synthesized. A structure-activity relationship study was performed using a variety of substituents to fine-tune the scaffold's potency. The installation of a methoxy group at the 5-position of the scaffold resulted in the discovery of compound 4.3.20, which exhibited the best activity with an EC50 of 3.6 ± 0.4 μM in rat L6 myoblasts and a half-life of 4.4 h in mice. Compound 4.3.20 displayed potential as an anti-obesity agent in a mouse model with an effective dose of 50 mg kg-1 without changes in food intake or lean mass. Tissue distribution studies revealed predominance in the liver and both white and brown adipose tissue. In addition, 4.3.20 improved serum markers of insulin sensitivity and hyperlipidemia such as insulin, glucose, triglycerides, cholesterol, and HOMA-IR. Taken together, compound 4.3.20 and related mitochondrial uncouplers show promise for further development in the treatment of obesity and other diseases. / Doctor of Philosophy / The mitochondria, which is an organelle within our cells, is where all the nutrients ingested in the form of food are metabolized, and either used for energy or stored as fat if they are not used. The latter is the main cause of obesity, carrying with it a myriad other comorbidities, such as high blood pressure, heart disease, diabetes, certain types of cancer. Obesity has become a great concern with an incidence of 42% in the US. Mitochondrial uncouplers are molecules that target the mitochondria with a mechanism of action of converting some of the energy ingested in the form of nutrients to be lost as heat instead of being stored as fat. The potential result is a regulated form of weight-loss. Herein, we developed a method for the synthesis of a novel mitochondrial uncoupler scaffold and disclose the mitochondrial uncoupler activity of over 150 molecules. In particular, compound 4.3.20 was tested in an obesity mouse model and was shown to induce fat loss with mice fed a high fat diet. Our investigations support potential use of mitochondrial uncouplers as a mechanism for the treatment and prevention of obesity and other metabolic diseases.

What is bearing management? It can be defined as follows¡G To achieve the best quality of the rotary machinery, the supervisor in charge shall possess the comprehensive knowledge, such as, to make the plan schedule and teamwork through the management method, to review the design and selection constantly, to do the problem analysis and the proper decision making in time. The purpose is to maintain the bearing in its good condition, and accordingly, to lower the emergency shut-down and man-power loss. At the year of 2011, CSC will encounter the strike of imbalanced structure of manpower caused by the trend of retirement. Since the consistent quality of maintenance is one of the enterprise¡¦s operation costs, and is also the foundation for the profit gaining and competition with others for survival, therefore, how to efficiently pass on and refine the technical expertise to cope with this foreseeable scenario becomes one the most importance challenges for CSC at this moment. To face the above-mentioned circumstances, the Plant Engineering & Maintenance Dept in CSC is determined to take up the bearing management as one of the crucial strategy. Some steps are adopted, such as, to set up the bearing seminars and establish the disciplinary knowledge through superior-subordinate communication and coordination, and meanwhile, to set up the benchmark to regulate the Bearing Consumption Rate (BCR), to regularly assess the use of bearing basing upon the tonnage of raw steel production, and then to execute the necessary methods to modify any possible problematic situation. The objective is clear, both in financial improvement and production efficiency. The target audiences of this research are from the members who completed the course in the seminars, they now also work as the front line personnel to facilitate in the field of bearing management. Meanwhile, some in-depth interviews with the maintenance superintendents are also applied. Some detections of the outcome are as follows¡G I. It is efficient to convey the bearing education and training via the events of bearing management. II. The support from the superintendent is the direct impact on the bearing management, and also is the vital factor to improve the efficiency. III. The content of curriculum for the training and education is another impact to the improvement of efficiency and success. IV. The lecturers must be qualified, capable and knowledgeable, and acknowledged by the fellow members. V. While the motive is clear and definite, the outcome can be much more enhanced. VI. Suitable location and environment with appropriate facilities are the basis requirement to set up the training program. According to the above-mentioned outcome, this research project has made some conclusions as suggested below¡G I. Staff of managerial level must fully support the bearing management and educational training. II. The empirical exemplification and the practical exercise are the main focus in the training progress, and shall also be the criterion as for the methodology in the other field of maintenance. III. To set a solid and objective goal to get a better result. IV. To set up an operational team to establish the task of the bearing maintenance training. V. The appropriate reward will be granted for any effective cases in the maintenance of bearing. VI. Launch the website of bearing maintenance, and broadly promote bearing maintenance and educational training via e-learning.

Evaluation of host geologic sediment interactions with carbon dioxide is very important in sequestration strategies. The objective of the study is to experimentally investigate the effects of different soil mineral types on carbon dioxide hydrate formation. At isothermal, isochoric, and isobaric conditions, batch experiments were conducted with different types of solids (bentonite, kaolinite, nontronite, pyrite, and soil) and electrolytes (NaCl, KCl, CaCl2, and MgCl2) to measure carbon dioxide hydrate formation times. A 50 mL pressurized vessel was used for the experiment by bubbling gaseous CO2 into the solid suspension. We observed that the formation time of carbon dioxide hydrate was dependent on the reactor temperature (273.4 K and 277.1 K) and types of solid and electrolyte. A clear peak was observed in the temperature profile of each experimental run and determined as the hydrate formation time. This is due to the initiation of the hydrate crystallization and latent heat release at the hydrate formation time. The temperature profiles vary significantly with respect to the types of solids and electrolytes. As crystallization initiates, peaks were observed at higher temperatures in pyrite and soil suspensions. The results showed that hydrate formation times for clay minerals in water were approximately twice and 10 times faster than that for pyrite and soil, respectively. The rates of gas consumption were able to be determined by the pressure monitoring. The kaolinite appeared to have the fastest gas consumption rate among the clay mineral suspensions, which was 2.4 times and 7.4 times faster than nontronite and bentonite, respectively. Results from these experiments seem to provide an insight on the formation and growth of carbon dioxide hydrate, once sequestered into the sea bed sediments under the deep sea environment.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22694号 / 医博第4638号 / 新制||医||1045(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 生田 宏一, 教授 竹内 理, 教授 濵﨑 洋子 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Sphingosine 1-phosphate (S1P) is a cellular signaling molecule that has been implicated in a variety of diseases including cancer, fibrosis, Alzheimer's, and sickle cell disease. It is formed from the phosphorylation of sphingosine (Sph) by sphingosine kinase (SphK) and SphK exists as two isoforms-"SphK1 and SphK2, which differ with respect to their cellular activity and localization. As the key mediators in the synthesis of S1P, SphKs have attracted attention as viable targets for pharmaceutical inhibition. To validate their potential as therapeutic targets, we aimed to develop potent, selective, and in vivo active inhibitors of SphK. Herein, we describe the design, synthesis and biological evaluation of SphK2 inhibitors. We first describe the development of six SphK2 inhibitors that assess the utility of replacing lipophilic tail groups with heterocyclic rings. These six compounds demonstrate that the lipid binding pocket for SphK2 cannot accommodate compounds with tail groups that are conformationally restricted or positively charged. We then describe the development of aminothiazole-based analogues of an SphK1-selective inhibitor. A library of 37 aryl-substituted aminothiazole tail groups were synthesized, revealing a structure-activity relationship study that examines electronic effects on the aryl-substituted aminothiazoles and the effect of modifying the amino portion of the aminothiazole. These molecules show surprisingly good potency and selectivity for SphK2. In particular, we highlight 3.20dd (SLC4101431), a biphenyl aminothiazole that is the post potent and selective SphK2 inhibitor to date, with an SphK2 Ki of 90 nM and 100-fold selectivity for SphK2. This molecule's in vivo activity will also be discussed. Mitochondrial uncouplers are small molecules that shuttle protons from the inter membrane space to the mitochondrial matrix independent of ATP synthase, which disrupts oxidative phosphorylation and promotes increased nutrient metabolism for homeostasis to be maintained. Consequently, small molecule mitochondrial uncouplers have been pursued as probes for mitochondrial function and as potential therapeutics for the treatment of obesity and type 2 diabetes. Herein, we describe the design, synthesis, and biological evaluation of small molecule mitochondrial uncouplers. We report a library of 52 compounds that have good mitochondrial uncoupling activity over a wide therapeutic range, including 5.16t (SHC4111522) and 5.17i (SHC4091665), which have EC50 values of 0.63 uM and 1.53 uM, respectively, and achieve at least 2-fold increase in oxygen consumption rates relative to basal levels. With these molecules, we demonstrate that pKa and cLogP significantly contribute to uncoupling activity and must be accounted for when developing new generation small molecule mitochondrial uncouplers. / Ph. D.

This study combines noninvasive hybrid diffuse optical spectroscopies [near-infrared spectroscopy (NIRS) and diffuse correlation spectroscopy (DCS)] with occlusive calibration for continuous measurement of absolute blood flow (BF), tissue blood oxygenation (StO2), and oxygen consumption rate (VO2) in exercising skeletal muscle. Subjects performed rhythmic dynamic handgrip exercise, while an optical probe connected to a hybrid NIRS/DCS flow-oximeter directly monitored oxy-, deoxy-, and total hemoglobin concentrations ([HbO2], [Hb], and [tHb]), StO2, relative BF (rBF), and relative VO2 (rVO2) in the forearm flexor muscles. Absolute baseline BF and VO2 were obtained through venous and arterial occlusions, respectively, and used to calibrate continuous relative parameters. Previously known problems with muscle fiber motion artifact in optical measurements were mitigated with a novel dynamometer-based gating algorithm. Nine healthy young subjects were measured and results validated against previous literature findings. Ten older subjects with fibromyalgia and thirteen age-matched healthy controls were then successfully measured to observe differences in hemodynamic and metabolic response to exercise. This study demonstrates a novel application of NIRS/DCS technology to simultaneously evaluate quantitative hemodynamic and metabolic parameters in exercising skeletal muscle. This method has broad application to research and clinical assessment of disease (e.g. peripheral vascular disease, fibromyalgia), treatment evaluation, and sports medicine.

This research investigates various applications of diffuse correlation spectroscopy (DCS) on in-vivo human muscle tissue, both at rest and during dynamic exercise. Previously suspected muscle tissue relative blood flow (rBF) baseline shift during extended measurement with DCS and DCS-Near infrared spectroscopy (NIRS) hybrid optical systems are verified, quantified, and resolved by redesign of optical probe and alteration in optical probe attachment methodology during 40 minute supine bed rest baseline measurements. We then translate previously developed occlusion techniques, whereby rBF and relative oxygen consumption rV̇O2 are calibrated to initial resting absolute values by use of a venous occlusion (VO) and arterial occlusion (AO) protocol, respectively, to the lower leg (gastrocnemius) and these blood flows are cross validated at rest by strain gauge venous plethysmography (SGVP). Methods used to continuously observe 0.5Hz, 30% maximum voluntary isometric contraction (MVIC) plantar flexion exercise via dynamometer are adapted for our hybrid DCS-Imagent diffuse optical flow-oximeter in the medial gastrocnemius. We obtain healthy control muscle tissue hemodynamic profiles for key parameters BF, V̇O2, oxygen saturation (StO2), deoxyhemoglobin, oxyhemoglobin, and total hemoglobin concentrations ([Hb], [HbO2], and THC respectively), as well as systemic mean arterial pressure (MAP) and pulse rate (PR), at rest, during VO/AO, during dynamic exercise and during 15 minute recovery periods. Next, we began investigation of muscle tissue hemodynamic disease states by performing a feasibility pilot study using limited numbers of controls and peripheral arterial disease (PAD) patients using the translated methods/techniques to determine the ability of our technology to assess differences in these populations.

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