Dissertations / Theses on the topic 'Animal Exchange'

In mammalian cells vesicle transport is an important process, many diseases are caused by defects of trafficking. Key elements for organelle trafficking are Rab proteins which regulate this system. Rabs are GTPases and belong to the Ras superfamily proteins. Rab27a is one of the main actors of this process, it promotes the recruitment of secretory vesicles at the release site, the plasma membrane, by its role of interconnection between the vesicle and the motor protein myosin V. Dysfunction leads to disease such as Griscelli syndrome. Rab27a is a molecular switch, in the active state is able to bind effectors. The switching from the GDP-bound to the GTP-bound is catalysed by Rab3GEP. Furthermore, to function properly Rab27a has to be targeted to the organelle membrane, this function has been attributed to the guanine exchange factor as well. Rab3GEP contains a DENN domain (differentially expressed in normal versus neoplastic) at the N-terminus and a death domain at the C-terminus. Rab3GEP belongs to the family of DENN domain proteins, in general DENN domains are considered as domains with a guanine exchange factor activity. Down-regulation of this Rab3GEP in melan-a melanocytes induces to melanosome clustering at the perinuclear area. There is no knowledge about the mechanism of Rab3GEP in the activation and targeting of Rab27a to the vesicle membranes. To better characterise Rab3GEP protein, truncations and point mutations of this GEF were performed and they were tested in cell based assays (using melanocytes with Rab3GEP knock out) and in pull down assays (using the capacity to activate Rab27a as a read out of the GEF activity). Moreover, Rab3GEP wild type, and some mutants were mis-targeted from the cytosol (normal localisation) to mitochondria, in order to investigate their capacity to rescue melanosome distribution in melan-Rab3GEP KO cells and to examine their ability to re-target Rab27a to a different membrane organelle. Results reported in this thesis indicate that point mutations within the DENN domain impair the ability of Rab3GEP to rescue the melanosome disperse distribution and to activate Rab27a in vitro assay, suggesting the crucial role of this domain for the Rab3GEP activity. However, the DENN domain alone is not sufficient to rescue melanosome distribution and to activate Rab27a. This suggests an essential role of the other parts of the protein. Moreover, evidence in this thesis indicate that the cytosolic localisation of Rab3GEP is essential for the ability of Rab3GEP to rescue melanosome distribution, and that the targeting activity of Rab3GEP towards Rab27a is dependent on its GEF activity. However, evidence obtained by studying melan-Rab3GEP KO cells indicate that despite the important role of Rab3GEP in Rab27a activation/targeting, it is not essential.

Chemical Exchange Saturation Transfer (CEST), is an emerging Magnetic Resonance Imaging (MRI) technique. CEST indirectly measures exchangeable protons contained in either endogenous or exogenous compounds by measuring the water signal reduction due to magnetisation exchange between these compounds and the surrounding water. CEST offers sensitivity enhancement compared to any method which directly measures these compounds. The complexity of the CEST signal in-vivo limits direct quantitative interpretation. However, the technique is inherently sensitive to a range of physiological parameters, such as temperature, pH and metabolite concentration. In order to investigate the relative importance of these different contributing factors, the work described in this thesis used the Bloch-McConnell equation system to model the CEST effect, for CEST sequence optimisation and data interpretation. Bovine Serum Albumin (BSA) phantoms were scanned with a CEST sequence and the results were compared to standard contrast methods (T1, T2). The CEST effects were correlated with changes in environmental pH, temperature and metabolite concentration. Next, a spectroscopic CEST sequence was implemented for spinal cord CEST and two models of neurodegenerative diseases were investigated. First, a model of Amyotrophic Lateral Sclerosis (ALS), revealed no changes in the CEST signal over the time course of the disease; the finding matched post-mortem soluble protein concentration analysis. Second, a model of Spinal and Bulbar Muscular Atrophy (SBMA), revealed no changes in the CEST signal of affected mice scanned at 10 and 12 months of age. However, changes in the CEST signal were observed in control mice and this again agreed with post-mortem protein concentration analysis. Finally, the potential for CEST to measure regional pH changes in a piglet model of Hypoxic Ischemia (HI) was investigated. CEST data were compared and found to agree with 31P MRS, measuring intracellular pH (pHi) and 1H MRS, measuring cerebral lactate levels.

[EN] For many years energy needs of ruminants have tried to be known to formulate rations adjusted, but it has been found that there are a variety of factors that affect them. Therefore, lots of studies are needed for evaluating the effect of these factors. Consequently, the main objective of this Thesis was to design and validate a respirometry system based on indirect calorimetry, which would allow assessing energy needs of small ruminants accurately. It was intended from the beginning it was a mobile system and of relatively low cost. Furthermore, a methane gas analyzer was incorporated to this system, which allowed the measurement of emissions of this greenhouse gas and quantification of energy losses in the form of methane. Initially the system had connected a mask, which was placed on the animal's face. A sample of exhaled gas was stored in a gas collection bag which was connected to the analyzer, and it measured the concentration of O2, CO2 and CH4 from the air. The proper functioning of the system was checked by a pilot experiment with dry Murciano-Granadina breed goats fed at maintenance level. Later this system was improved. Some of the most important changes were the replacement of the mask by a head hood in which the animal introduced the whole head, and the development of software that recorded and kept automatically concentrations of O2, CO2 and CH4 in exhaled air. This improvement allowed gas measurements during longer periods of time and recording more data. These changes were also validated through a pilot test with dry Manchega breed sheep. Subsequently, three experiments were performed. One of them with dry Guirra ewes and the other two with Murciano-Granadina goats during mid lactation. Diets were mixed rations that differed in the inclusion of cereal or fibrous by-products. In these experiments the effect of diet was studied on digestibility, energy balance and carbon-nitrogen, oxidation of nutrients, rumen parameters and methane production; in the case of lactating goats, also on milk performance. The determination of the calibration factor for O2 (1.005 ± 0.0101) confirmed the proper functioning of equipment. Moreover, small differences between the heat production obtained by indirect calorimetry and the carbon-nitrogen balance (2% in sheep and 1% in goats) demonstrated that this system allows determining the heat production of the animals reliably and accurately. In the experiments of this Thesis have been estimated maintenance energy needs of two Spanish native sheep breeds, such as the sheep from the Guirra and Manchega breeds; net maintenance requirements were 270 kJ/kg BW0.75, on average. In the case of Murciano-Granadina breed goats, in the middle of lactation, the average utilization efficiency of metabolizable energy for lactation was 0.61.
[ES] Desde hace años se ha tratado de conocer las necesidades energéticas de los rumiantes con el fin de formular raciones ajustadas, pero se ha comprobado que hay una gran variedad de factores que les afectan; por ello son necesarios estudios que evalúen el efecto de estos factores. Como consecuencia, el principal objetivo de esta tesis fue diseñar y validar un equipo de respirometría, basado en calorimetría indirecta, que permitiese evaluar las necesidades en energía de pequeños rumiantes de forma precisa. Se pretendió desde el inicio que fuese un sistema móvil y de relativo bajo coste. Además, a este sistema también se le incorporó un analizador de gas metano, que permitía la medición de las emisiones de este gas de efecto invernadero y la cuantificación de las pérdidas energéticas en forma de metano. Inicialmente el equipo tenía conectada una máscara que se colocaba en la cara del animal. Una muestra del gas espirado era almacenada en una bolsa de recogida de gases que era conectada al analizador, el cual medía la concentración de O2, CO2 y CH4 del aire. Se comprobó el correcto funcionamiento del sistema mediante una prueba piloto con cabras de raza Murciano-Granadina secas, alimentadas a nivel de mantenimiento. Posteriormente este sistema fue mejorado. Algunos de los cambios más importantes fueron la sustitución de la máscara por una urna en la que el animal introducía la cabeza entera, y el desarrollo de un software que registraba y guardaba de forma automática las concentraciones de O2, CO2 y CH4 del aire espirado. Esta mejora permitía medidas de gases durante periodos de tiempo más largos y el registro de muchos más datos. Estas modificaciones también fueron validadas mediante una prueba piloto con ovejas de raza Manchega secas. Posteriormente se realizaron tres experimentos. Uno de ellos con ovejas de raza Guirra secas y los otros dos con cabras Murciano-Granadinas en mitad de lactación. Las dietas fueron raciones mixtas que diferían en la inclusión de cereal o subproductos fibrosos. En estos experimentos se estudió el efecto de la dieta sobre la digestibilidad, balances de energía y carbono-nitrógeno, oxidación de los nutrientes, parámetros del rumen y producción de metano; en el caso de las cabras en lactación, también sobre los rendimientos productivos. La determinación del factor de calibrado para el O2 (1,005 ± 0,0101) confirmó el buen funcionamiento del equipo. Por otro lado, las pequeñas diferencias entre la producción de calor obtenida mediante calorimetría indirecta y el balance de carbono-nitrógeno (2% en ovejas y 1% en cabras) demostraron que este sistema permite determinar la producción de calor de los animales de forma fiable y precisa. En los trabajos de esta Tesis se han estimado las necesidades energéticas de mantenimiento en dos razas de ovejas autóctonas españolas, como son las razas Guirra y Manchega; las necesidades netas de mantenimiento fueron 270 kJ/kg PV0,75, de media. En el caso del ganado caprino de raza Murciano-Granadina, en mitad de lactación, la eficacia media de utilización de la energía metabolizable para la lactación fue de 0,61.
[CAT] Des de fa anys s'ha tractat de conèixer les necessitats energètiques dels remugants a fi de formular racions ajustades, però s'ha comprovat que hi ha una gran varietat de factors que els afecten; per això són necessaris estudis que avaluen l'efecte d'estos factors. Com a conseqüència, el principal objectiu d'aquesta Tesi va ser dissenyar i validar un equip de respirometría, basat en calorimetria indirecta, que permetera avaluar les necessitats en energia de menuts remugants de forma precisa. Es va pretendre des de l'inici que fóra un sistema mòbil i de relatiu baix cost. A més, a este sistema també se li va incorporar un analitzador de gas metà, que permetia el mesurament de les emissions d'este gas d'efecte hivernacle i la quantificació de les pèrdues energètiques en forma de metà. Inicialment l'equip tenia connectada una màscara que es col·locava en la cara de l'animal. Una mostra del gas expirat era emmagatzemada en una bossa d'arreplega de gasos que era connectada a l'analitzador, el qual mesurava la concentració d'O2, CO2 i CH4 de l'aire. Es va comprovar el funcionament correcte del sistema per mitjà d'una prova pilot amb cabres de raça Murciano-Granadina seques, alimentades a nivell de manteniment. Posteriorment este sistema va ser millorat. Alguns dels canvis més importants van ser la substitució de la màscara per una urna en què l'animal introduïa el cap sencera, i el desenrotllament d'un programari que registrava i guardava de forma automàtica les concentracions d'O2, CO2 i CH4 de l'aire expirat. Esta millora permetia mesures de gasos durant períodes de temps més llargs i el registre de moltes més dades. Estes modificacions també van ser validades per mitjà d'una prova pilot amb ovelles de raça Manxega seques. Després es van realitzar tres experiments. Un d'ells amb ovelles de raça Guirra seques i els altres dos amb cabres Murciano-Granadinas en mitat de lactació. Les dietes van ser racions mixtes que diferien en la inclusió de cereal o subproductes fibrosos. En estos experiments es va estudiar l'efecte de la dieta sobre la digestibilitat, balanços d'energia i carboni-nitrogen, oxidació dels nutrients, paràmetres del rumen i producció de metà; en el cas de les cabres en lactació, també sobre els rendiments productius. La determinació del factor de calibrat per a l'O2 (1,005 ± 0,0101) va confirmar el bon funcionament de l'equip. D'altra banda, les xicotetes diferències entre la producció de calor obtinguda per mitjà de calorimetria indirecta i el balanç de carboni-nitrogen (2% en ovelles i 1% en cabres) van demostrar que este sistema permet determinar la producció de calor dels animals de forma fiable i precisa. En els treballs d'esta Tesi s'han estimat les necessitats energètiques de manteniment en dos races d'ovelles autòctones espanyoles, com són les races Guirra i Manxega; les necessitats netes de manteniment van ser 270 kJ/kg PV0,75, de mitja. En el cas del bestiar caprí de raça Murciano-Granadina, en mitat de lactació, l'eficàcia mitjana d'utilització de l'energia metabolitzable per a la lactació va ser de 0,61.
López Luján, MDC. (2015). Development of a mobile open-circuit system based on indirect calorimetry for energetic metabolism studies in small ruminants [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/50430
TESIS

[EN] The main objective of this Thesis was to study the energy metabolism in small ruminants under different nutrition sceneries. As methodology we utilized indirect calorimetry instead of direct calorimetry or feeding trials. Within indirect calorimetry we worked with a portable open circuit gas exchange system with a head hood. This open circuit respiration system permitted completed the whole energy balance and evaluate the efficiency of utilization of the energy of the diet for different physiological circumstances as milk production in dairy goats or maintenance in sheep. Besides, we could quantify in each trial some of the wastes related to environmental pollution as CH4 emissions and excretion of nitrogen in feces and urine. In this thesis three experiments were designed, two in dairy goats and other in sheep as we described below. In the first experiment, digestibility, energy balance, carbon and nitrogen balance, milk performance, rumen parameters and milk fatty acids and metabolites were obtained. Metabolic cages and open circuit indirect calorimetry system were the methods applied. Treatments consist in two mixed diets with alfalfa as forage. Within the concentrate, oat grain was replaced with rice bran. No significant differences were found for metabolizable energy intake (MEI), 1254kJ/kg of BW0.75 on average, and heat production (HP); 640 kJ/kg of BW0.75 on average. No differences were obtained for milk production (2.2 kg/d on average) and milk fat was greater in the rice bran diet (6.9% vs. 5.3% for rice bran and oat diets, respectively). Energy balance was positive and milk metabolites correlated these found. Regards to CH4 emissions, determined in vivo by gas exchange indirect calorimetry, goats fed the rice bran significantly reduced methane production (23.2 g/d vs 30.1 g/d). In the second experiment, we also used two types of diets, but in this case we substituted the forage and maintained the same concentrate; in one of the diets a grass (Maralfafa [Pennisetum sp.]- M diet) was used as forage and in the other an extensively used leguminous (Alfalfa [Medicago sativa]- A diet). Methods and analysis were the same that in Experiment 1, and here we include metabolites in urine and blood plasma. The dry matter intake was higher for diet A (1.8 vs 1.6 kg/d, respectively) and digestibility coefficients were higher for diet M. However, no significant differences were shown in MEI (1089 kJ/kg of BW0.75, on average) and HP (639 kJ/kg of BW0.75on average). Higher milk yield was observed in A diet than M diet (1.8 vs. 1.7 kg/d, respectively) and metabolites in urine, plasma and milk indicated better use of diet A than M, while no differences in milk composition were found (5% of fat and 4.3% of protein). Methane production was higher for A diet (28.5 g/d) than M diet (25.9 g/d), although these differences were not statistically significant. In the third experiment, energy partition was compared in two sheep breeds (Manchega vs. Guirra) fed above maintenance. We fed again with mixed diets and metabolic cages, indirect calorimetry, nitrogen balance and integral calculus were the tools used for these energy partitioning approach. An approximation of division of heat production was done. ME for maintenence was estimated at 354 kJ/kg of BW0.75and day, on average for the two breeds. Basal metabolic rate was different between breeds; 270 vs. 247 kJ/kg of BW0.75for Guirra and Manchega, respectively.
[ES] El objetivo principal de esta tesis fue estudiar el metabolismo energético en pequeños rumiantes bajo diferentes escenarios de nutrición. Como metodología se utilizó la calorimetría indirecta en vez de calorimetría directa o pruebas de alimentación. Dentro de la calorimetría indirecta trabajamos con un sistema portátil de circuito abierto de intercambio de gases con una "urna" (Heat hood). Este sistema de circuito abierto de respiración nos permitió completar todo el balance energético y evaluar la eficiencia de la utilización de la energía de la dieta para diferentes estados fisiológicos como producción de leche en cabras u oveja en mantenimiento. Además fue posible cuantificar en cada ensayo algunas perdidas relacionadas con la contaminación ambiental como emisiones de CH4 y la excreción de nitrógeno en heces y orina. En esta tesis se diseñaron tres experimentos, dos en cabras en lactación y otro en ovejas como describimos a continuación. En el primer experimento se han determinado, digestibilidad, balance energético, balance carbono nitrógeno, producción de leche, parámetros ruminales, ácidos grasos y metabolitos en leche. Jaulas metabólicas y un sistema de circuito abierto de calorimetría indirecta fue el método aplicado. Los tratamientos consistieron en dos dietas mixtas con alfalfa como forraje y dentro del concentrado el grano de avena fue reemplazado por cilindro de arroz. No se encontraron diferencias significativas en la energía metabolizable ingerida (MEI) de 1254 kJ/kg PV0.75 en promedio y una producción de calor (HP) de 640 kJ/kg PV0.75 en promedio. La producción de leche no presentó diferencias significativas entre las dos dietas, (2,2 kg/den promedio), la grasa de la leche fue mayor en la dieta de cilindro de arroz (6,9% vs. 5,3% para cilindro de arroz y avena respectivamente). El balance energético fue positivo y correlacionado a los metabolitos en leche determinados. En cuanto a las emisiones de CH4, determinadas en vivo mediante el intercambio de gases por calorimetría indirecta, las cabras alimentadas con el subproducto redujeron significativamente la producción de metano (23,2 g / d vs 30,1 g / d.). En el segundo experimento, también utilizamos dos tipos de dietas, pero en este caso sustituimos los forrajes y mantuvimos el mismo pienso; en una de las dietas se utilizó como forraje una gramínea (Maralfafa [Pennisetum sp.] - dieta M) y en el otro una leguminosa de uso extendido (Alfalfa [Medicago sativa] - dieta A). Los métodos de análisis y análisis fueron los mismos que los utilizados en el Experimento 1, y se incluyeron además análisis de metabolitos en orina y plasma. La materia seca ingerida fue mayor para dieta A (1,8 vs 1,6 kg/d, respectivamente), los coeficientes de digestibilidad fueron mayores para la dieta M. Sin embargo, no se encontraron diferencias significativas en MEI (1089 kJ/kg PV0.75, en promedio) y HP 639 kJ/kg PV0.75, en promedio). La producción de leche fue mayor en la dieta A que la dieta M, (1,8 vs. 1,7 kg/d, respectively) y los metabolitos en orina, plasma y leche indican un mejor aprovechamiento de la dieta A. No se presentaron diferencias en la composición de la leche (5% de grasa and 4.3% de proteína). La producción de metano fue mayor para la dieta A (28,5 g/d) que para la dieta M (25,9 g/d), aunque estas diferencias no fueron estadísticamente significativas. En el tercer experimento se compararon la partición energética en dos razas de ovejas (Manchega vs. Guirra) en mantenimiento. Fueron alimentadas con dietas mixtas en jaulas metabólicas, calorimetría indirecta, balance carbono nitrógeno y cálculos integrales fueron las herramientas utilizadas para un aproximación de la partición energética. Se realizó una aproximación de división de producción de calor. El ME para mantenimiento se estimó en 354 kJ/kg PV0.75/ día, en promedio para las dos razas. Las diferencias en la tasa metabólica basal entre las razas fu
[CAT] El principal objectiu d'aquesta tesi va ser estudiar el metabolism energètic en xicotets ruminants baix diferents escenaris de nutrició. Com a metodologia es va utilitzar la calorimetria indirecta en compte de calorimetria directa o proves d'alimentació. Dins de la calorimetria indirecta treballarem amb un sistema portatil de circuit obert d'intercanvi de gasos amb "urna" (Heat hood). Aquest sistema de respiració de circuit obert ens va permetre completar tot el balanç energètic i avaluar l'eficiència de la utilització de l'energia de la dieta per a diferents circumstàncies fisiològiques com produccion de llet en cabres o manteniment en ovelles. A més va ser possible quantificar en cada assaig algunes perdues relacionades amb la contaminacion ambiental com a emissions de CH4 i l'excreció de nitrogen en femta i orina. En aquesta tesi es van dissenyar tres experiments, dos en cabres en lactación i un altre en ovelles com vam descriure a continuació. En el primer experiment s'han determinat,digestibilidad, balanç energètic, balanç carboni nitrogen, producció de llet, paràmetres ruminales, àcids grassos i metabòlits en llet. Gàbies metabòliques i un sistema de circuit obert de calorimetria indirecta va ser el mètode aplicat. Els tractaments van consistir en dues dietes mixtes amb alfals com a farratge i dins del concentrat el gra de civada va ser reemplaçat per cilindre d'arròs. No es van trobar diferències significatives en l'energia metabolizable ingerida (MEI) de 1254 kJ/kg PV0.75 en mitjana i una producció de calor (HP) de 640 kJ/kg PV0.75 en mitjana. La producció de llet no va presentar diferències significatives entre les dues dietes, (2.2 kg/donen mitjana), el greix de la llet va ser major en la dieta de cilindre d'arròs (6.9% vs. 5.3% per a cilindre d'arròs i civada respectivament). El balanç energètic va ser positiu i correlacionat als metabòlits en llet determinats. Quant a les emissions de CH4, determinades en viu mitjançant l'intercanvi de gasos per calorimetria indirecta, les cabres alimentades amb el subproducte van reduir significativament la producció de metà (23.2 g / d vs 30.1 g / d.).En el segon experiment, també utilitzem dos tipus de dietes, però en aquest cas substituïm els farratges i vam mantenir el mateix pinso; en una de les dietes es va utilitzar com a farratge una gramínea (Maralfafa [Pennisetum sp.] - dieta M) i en l'altre una **leguminosa d'ús estès (Alfals [Medicago sativa] - dieta A). Els mètodes d'anàlisis i anàlisis van ser els mateixos que els utilitzats en l'Experiment 1, i es van incloure a més anàlisi de metabòlits en orina i plasma. La matèria seca ingerida va ser major per a dieta A (1,8 vs 1,6 kg/d, respectivament), els coeficients de digestibilidad van ser majors per a la dieta M. No obstant açò no es van trobar diferències significatives en MEI (1089 kJ/kg PV0.75, en mitjana) i HP 639 kJ/kg PV0.75, en mitjana). La producció de llet va ser major en la dieta Al fet que la dieta M, (1,8 vs. 1,7 kg/d, respectively) i els metabòlits en orina, plasma i llet indiquen un millor aprofitament de la dieta A. No es van presentar diferències en la composició de la llet (5% de greix i 4.3% de proteïna). La producció de metà va ser major per a la dieta A (28,5 g/d) que per a la dieta M (25,9 g/d), encara que aquestes diferències no van anar estadísticament significatives. En el tercer experiment es van comparar la partició energètica en dues races d'ovelles (Manxega vs. Guirra) en manteniment. Van ser alimentades amb dietes mixtes en gàbies metabòliques, calorimetria indirecta, balanç carboni nitrogen i càlculs integrals van ser les eines utilitzades per a un aproximació de la partició energètica. Es va realitzar una aproximació de divisió de producció de calor. L'EM para manteniment es va estimar en 354 kJ/kg PV0.75 / dia, en mitjana per a les dues races. Les diferències en la taxa metabòlica basal entre les races va ser de 2
Criscioni Ferreira, PF. (2016). Application of an open circuit indirect calorimetry system for gaseous exchange measurements in small ruminant nutrition [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/64069
TESIS

Ion exchange resin capsules provide a possible alternative to conventional soil testing procedures. Previous studies with semi-arid, low fertility soils observed poor relationships with poorly mobile nutrients such as phosphorus (P). We propose that placement depth may improve those relationships. Our objective was to (1) determine if placement depth could improve resin capsule estimation of the bioavailability of nitrogen (N), P, and sulfur (S) and (2) to determine if resin capsules can effectively estimate S availability in semi-arid, low fertility soils. Field sites were established in Rush and Skull Valleys, Utah on loam and sandy loam soils, respectively. Fertilizer was surface applied as ammonium sulfate and triple superphosphate with six N, P and S treatments (0, 5.5, 11, 22, 44 and 88 kg ha-1 of N and P2O5 and 0, 7, 14, 28, 56 and 112 kg ha-1 of S). Thirty 4.0-m2 plots were established at each field location. Resin capsules were placed three per plot at 0–5, 5–10, and 10–15 cm deep in the soil and soil samples taken at respective depths. The capsules were removed and replaced after approximately 90 d. Final removal and soil sampling occurred approximately 240 d later. For the second study, fertilizer was surface applied as ammonium sulfate with six S treatments (0, 7, 14, 28, 56 and 112 kg ha-1 of S) with one resin capsule placed in each 4.0-m2 plot at a depth of 5 cm in the soil. Resin capsules were removed and replaced approximately every 90 d for a total of four samplings. Soil samples were taken with every resin capsules install and removal. In the first study, bicarbonate extractable P was significantly related to P application at all depths and times except the two lowest depths at the time of final sampling, and resin capsule P was only related to P application 398 days after application in the 0–5 and 5–10 cm depths. However, this is an improvement in estimates of bioavailability compared to a single placement depth. The 5–10 cm depth was the best for placement for determination of NH4-N, and resin capsules improved upon soil test estimates. For NO3-N, depth was not important, but resin capsules had a stronger relationship with N applied than the soil test 398 d after application. In addition, both resin capsules and the S soil test were related to S applied, but resin capsules were more able to pick up S cycling through the system. In the second study resin capsules and conventional soil tests were both effective in distinguishing between fertilizer rates, though only the conventional soil test was related to S applied at the last sampling (366 d after fertilizer application). Overall resin capsules were effective at reflecting application rates, and may be a good tool to estimate nutrient bioavailability. Correlation with plant uptake is required to determine if soil tests or resin capsules were a better estimate of bioavailable nutrients.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv.
Bilagan utgöres av sammanfattnig på svenska med titeln: Fysiologiskt svar på arbete hos varmblodiga travhästar i relation till total blodvolym. Härtill 5 uppsatser.

Swine production represents approximately 40% of the world's meat production, and swine wastes contain high concentrations of organic matter, nitrogen (N) and phosphorus (P). Swine production is intensifying as meat demand increases and concentrated animal feeding operations (CAFOs) are becoming increasingly common, making it difficult to treat the waste generated. A system for holistic treatment of swine waste produced in CAFOs was investigated in this study that sustainably generates energy and recovers N and P as saleable fertilizers. The system uses anaerobic digestion (AD) for methane production and solids stabilization, followed by precipitation of struvite (MgNH4PO4*6H2O) and recovery of N by ion exchange onto natural zeolites. This process is expected to mitigate both eutrophication of receiving waters and greenhouse-gas emissions while generating products that meet agronomic nutrient demands; however, the economic and environmental sustainability remains unknown. The objectives of this study were to: (1) evaluate water quality and the fate of nutrients and ions in each step in the proposed system through pilot and bench scale experiments, (2) evaluate content/quality of struvite precipitates formed in wastewater treatment processes, (3) assess basic composition of zeolite materials that are being considered for use as IX materials, (4) quantify the environmental impact of the proposed system, and (5) estimate the economic benefits and costs of the proposed system. The results of a bench scale evaluation of the system show that although water quality greatly improves throughout the treatment process, the effluent water quality has high concentrations of COD (2,803 mg O2/L) and E. coli (106.3 CFU/100ml). This limits reuse options for the reclaimed water, however a variety of on-farm applications may be suitable. During struvite precipitation, the recovery efficiency of SRP was 87% (60 mg/L recovered); however, although measurements that take into account P in suspended solids show a lower recovery efficiency, they also show higher mass recovery (77% efficiency, 66 mg/L recovered). N recovery during struvite precipitation showed a similar trend, with 49% of TN and 7% of NH4-N being recovered. Struvite recovery can only occur from NH4-N and soluble reactive P. The additional recovery observed is likely due to adsorption of the nutrients onto the precipitate. Therefore, to accurately measure and report recovery, measurements of N and P that take into account suspended solids should be used. In most wastes, magnesium is the limiting constituent for struvite formation, but for swine AD effluents, P is the limiting constituent. Therefore, a higher soluble P concentration would increase recovery potential. The majority of the remaining N and P as well as a significant amount of potassium (K) were recovered during IX. Six struvites from commercial processes as well as our bench-scale experiments were assessed and compared by X-ray diffraction, SEM imaging, and SEM-EDX scans. All samples were confirmed as struvite by XRD, however they varied widely in crystal size and shape. The elemental composition of the samples was similar; however, struvite formed from phosphate mining waste had higher amounts Mg and P, indicating more pure struvite formation. The presence of impurities in some samples was likely due to the reactor design and solids separation methods. XRD was also used to confirm the identity of zeolites. Three clinoptilolites had similar crystal size and elemental composition except for Zeosand [reg] which showed a surface roughness, which likely contributes to higher cation exchange capacity. Chabazite has smaller crystal size and larger pores than clinoptilolite, which also likely contributes to its higher capacity. Life cycle assessment (LCA) was used to evaluate the environmental sustainability of the system and the results suggested that environmental benefits were provided across almost all impact categories. Two alternatives for raising the pH in struvite precipitation (NaOH addition vs. aeration) and two alternatives for zeolite IX materials (chabazite vs. clinoptilolite) were assessed, but there were negligible differences between alternatives. The system was also assessed at a medium and large scale, and the large scale was more environmentally friendly across all categories. Operational impacts were significantly greater than construction impacts; therefore, the environmental impact of the system can be accurately assessed by only including operation. A life cycle cost assessment (LCCA) was also performed on the system and showed a payback period of 39 years for a medium sized system and 15 years for a large size. This, however, is when compared to a "business-as-usual" scenario and does not consider renewable energy credits or government grants. Furthermore, although a larger system is more economically beneficial, this must be balanced with quality of animal care. From a cost standpoint, IX recovery using chabazite is not recommended and struvite precipitation using aeration is more economically beneficial than NaOH addition.

Thesis(M.A.)--Case Western Reserve University, 2010
Title from PDF (viewed on 2010-01-28) Department of Communication Sciences Includes abstract Includes bibliographical references and appendices Available online via the OhioLINK ETD Center

The purpose of this work is to answer the basic question, if the economics can explore the animal behaviour. At first the determination of the subject of economics is needed. It means to point out, what should economics deal with. I will come to the point of the problem of purposeful action, the problem of rationality and exchange. Consequently I will explore, if it is possible to apply this theory to animal behaviour and if such an application has a sense. I will advert to the problems, what are made by the economists in their opinions of the animal behaviour and on argumentation why the animal behaviour is not a subject of economics.

Plasma membrane Na+/H+ exchangers (NHEs) have been shown to be inhibited by amiloride and its derivatives. Studies have demonstrated that these antagonists act on the transporter by interacting with amino acid residues within certain transmembrane domains (TM4 and 9). Experiments in our laboratory have identified three novel sites involved in inhibitor interactions (PP157--8, E350, and G356). The general focus of this thesis was to study the structure-function relationship of NHE1. One of the aims of this thesis was to assess whether mutations at these novel sites also affected transport kinetics (i.e., Na+ and H+ affinity). The results showed that neither Na+ nor H+ affinities were significantly altered with any of the mutations analyzed. The next objective was to investigate the nature of the interaction between the exchanger and pharmacological agents, by measuring transport activity in the presence of substituted guanidinium antagonists. The results suggest an interaction between L167 and the chlorine moiety at position 3 of the benzoyl group of a novel benzoyl guanidinium compound, while G356 appears to interact with the chlorine moiety at position 4 (rather than position 3) of the benzene ring.
The last objective was to define the membrane topology of NHE1. The methodology involves reintroducing cysteine residues into a cysteine-less mutant NHE1 and assessing their accessibility with thiol-reactive agents. Unfortunately, these results were inconclusive and further optimization of the assay conditions is required.

Sodium/proton exchangers (NHEs) are secondary active transporters that catalyze the electroneutral exchange of Na+ and H+ down their respective concentration gradients using the inward electrochemical Na+ gradient established by the plasma membrane Na+ /K+-ATPase. NHEs play a fundamental role in maintaining the intracellular pH (pHi) homeostasis and cell volume which are required for stable protein activity, and ultimately cell survival. To date, nine NHE isoforms (NHE 1-9) have been identified, and are expressed in a tissue/cell specific manner, which indicates a degree of specialization in their respective cellular functions. NHE1 through 5 are plasmalemmal-type exchangers, whereas NHE6 and 7 accumulate predominantly in organellar compartments. Substantially less information is known about NHE8 and NHE9. Initial reports have shown that both NHE8 and NHE9 mRNA are ubiquitously expressed, and that NHE8 protein can be detected in the proximal tubules of kidney nephrons (Goyal et al., 2003; de Silva et al., 2003).
To gain further insight into the subcellular distribution and function of these latter isoforms, full length cDNAs of NHE8 and NHE9 were cloned, epitope tagged and transfected into cultured cell lines. Examination of their subcellular localization by confocal microscopy showed that NHE8 and NHE9 are targeted to lysosomes and endosomes respectively. Furthermore, co-immunoprecipitation experiments indicated that NHE9 can form homodimeric complexes. Co-immunoprecipitation studies and confocal microscopy also indicated that NHE9 may form heterocomplexes with NHE8. In order to further characterize NHE9 under native conditions, a native polyclonal antibody was raised against a distinct region of its C-terminal cytoplasmic tail.

Na+/H+ exchanger (NHE) is a major contributor in controlling intracellular pH. The activity of this protein is allosterically modified by intracellular H+. Histidine residues of the NHE that face the cytoplasm may be involved in determining the intracellular pH set point, with their state of protonation influencing the rate of Na +/H+ exchange. To test this hypothesis, histidine residues in the ubiquitously expressed NHE isoform (NHE1) that are relatively conserved amongst members of the NHE gene family were substituted by site-directed mutagenesis and the mutants were stably transfected into mammalian cells that are deficient in endogenous Na+/H + exchange activity. The pHi sensitivity of each mutant was evaluated by measuring the rate of 22Na + influx as a function of the intracellular H+ concentration. Mutation of the histidines located at the putative cytoplasmic face of the N-terminal transmembraneous domain of NHE1 did not show any significant effect on the pHi sensitivity of the protein. By contrast, substitution of histidines located in the C-terminal cytoplasmic tail activated the exchanger by increasing its sensitivity to H+. These mutants were no longer activated in response to protein kinase C, when compared to wild type. Taken together, these data support the hypothesis that some of the relatively conserved histidine residues in the C-terminal cytoplasmic tail of NHE1 may be involved in determining the pHi "threshold" or "set point" of the transporter.

The Na+/H+ exchangers (NHE) mediate the electroneutral exchange of sodium for protons and play integral roles in sodium, acid-base and cell volume homeostasis. Presently, eight isoforms of the NHE (NHE1 to NHE8) have been identified that are targeted to distinct membrane compartments. The focus of this study is to characterize in greater detail the biosynthesis and differential sorting of two closely related organellar NHE isoforms, NHE6 and NHE7. Previous studies have established that NHE7 accumulates in the trans-Golgi network and associated endosomes, whereas the localization of NHE6 remains controversial. In one study, HeLa cells transiently expressing low levels of a green fluorescent protein-tagged construct of NHE6 showed close co-localization with mitochondrion-specific dyes. However, when NHE6 was overexpressed in COS7 cells, significant accumulation was observed throughout the cell in membrane vesicles derived from the endoplasmic reticulum. To further address this discrepancy, NHE6 engineered to contain the influenza virus hemagglutinin (HA) epitope at its C-terminus (NHE6HA), was subcloned into the ecdysone-inducible expression vector pIND and stably transfected into Chinese hamster ovary cells that constitutively express the ecdysone receptor. NHE6HA expression was stimulated by ponasterone A, an ecdysone analogue. The localization of NHE6 was determined biochemically by subcellular fractionation of cell lysates and visually by immunofluorescence confocal microscopy of intact cells using antibodies that recognize the HA-epitope and various organellar specific markers. Similar studies were conducted with an NHE7-inducible mammalian expression system. Our findings indicate that NHE6 is differentially processed through distinct Golgi-dependent and -independent pathways, ultimately accumulating in recycling endosomal vesicles. Little evidence was found to support sorting to mitochondria. Furthermore, we show that NHE6 is synthesiz

Na+/H+ exchangers (NHE) are transmembrane proteins that mediate the electroneutral exchange of Na+ and proton across the plasma membrane. There are seven mammalian isoforms of the gene family identified to date. The recently cloned isoform NHE5 seems to be the most highly cell type-specific being probably expressed only in neurons. The objective of my research was to describe the basic biochemical and some of the regulatory characteristics of NHE5 in an effort to understand its in vivo function. To this end, the full-length cDNA of NHE5 was reconstructed and transfected into the NHE-deficient Chinese hamster ovary cell line AP1.
Pharmacological analyses demonstrated that H+ i-activated 22Na+ influx mediated by NHE5 was inhibited by several classes of drugs at half-maximal concentrations that were intermediate to those determined for the high-affinity NHE1 and the low-affinity NHE3 isoforms. Kinetic analyses showed that the extracellular Na+-dependence of NHE5 activity followed a simple hyperbolic relationship and, unlike other NHE isoforms, the intracellular H+-dependence also exhibited first-order kinetics. Extracellular monovalent cations, such as H+ and Li+, but not K+, acted as effective competitive inhibitors of 22Na+ influx by NHE5.
To find novel interacting proteins that are involved in NHE5 regulation, a yeast two-hybrid screen of human brain cDNA library was conducted using NHE5 as bait. A clone encoding the AMP-activated protein kinase (AMPK) alpha2 subunit was further analyzed. AMPK is a serine/threonine kinase that is activated by elevated ratios of [AMP]/[ATP], regulating various biological processes in response to hypoxia or exercise. AMPK alpha2 binds NHE5 in vitro and in vivo, and directly phosphorylates it in vitro. Activation of endogenous AMPK by AICAR, a membrane permeable AMP analogue, as well as heterologous expression of the full-length and constitutive active forms of alpha2 subunit increased the transporter activity measured by 22Na+ influx.
The regulatory protein arrestin3 was also found to interact with NHE5 in the yeast two-hybrid screen. Arrestins were previously shown to associate with and regulate transmembrane proteins of the G protein-coupled receptor family. We demonstrate that NHE5 binds arrestin3 both in vitro and in vivo; and the binding is phosphorylation-dependent. When co-expressed in CHO cells, arrestin3 and NHE5 co-localize, and arrestin3 expression seems to attenuate the basal activity of the transporter. The data presented in this thesis reveals new aspects of both NHE regulation, and AMPK and arrestin function.

Cerebral ischaemia is a leading cause of disability and death world-wide. The only effective treatments are thrombolytic therapy (plasminogen activator; tPA) and hypothermia (33?C). However, tPA has limited clinical application due to its short therapeutic time window and its specific application in thrombo-embolic stroke. Moderate hypothermia (33?C) is only being used following cardiac arrest in comatose survivors. Hence more treatments are urgently required. The first step in developing new treatments is the identification and characterisation of a potential therapeutic target. Since brain damage following cerebral ischaemia is associated with disturbances in intracellular calcium homeostasis, the sodium-calcium exchanger (NCX) is a potential therapeutic target due to its ability to regulate intracellular calcium. Currently, however there is uncertainty as to whether the plasma membrane NCX has a neuroprotective or neurodamaging role following cerebral ischemia. To address this issue I compared hippocampal neuronal injury in NCX3 knockout mice (Ncx3-/-) and wild-type mice (Ncx3+/+) following global cerebral ischaemia. In order to perform this study I first established a bilateral common carotid occlusion (BCCAO) model of global ischaemia in wild-type C57/BlHsnD mice using controlled ventilation. After trials of several ischaemic time points, 17 minutes was established as the optimum duration of ischaemia to produce selective hippocampal CA1 neuronal loss in the wild-type mice. I then subjected NCX3 knockout and wild-type mice to 17 minutes of ischaemia. Following the 17 minute period of ischaemia, wild-type mice exhibited 80% CA1 neuronal loss and 40% CA2 neuronal loss. In contrast, NCX3 knockout mice displayed > 95% CA1 neuronal loss and 95% CA2 neuronal loss. Following experiments using a 17 minute duration of global ischaemia, a 15 minute duration of ischaemia was also evaluated. Wild-type mice exposed to a 15 minute period of ischaemia, did not exhibit any significant hippocampal neuronal loss. In contrast, NCX3 knockout mice displayed 45% CA1 neuronal loss and 25% CA2 neuronal loss. The results clearly demonstrate that mice deficient for the NCX3 protein are more susceptible to global cerebral ischaemia than wild-type mice. My findings showing a neuroprotective role for NCX3 following ischaemia, suggest that the exchanger has a positive role in maintaining neuronal intracellular calcium homeostasis. When this function is disrupted, neurons are more susceptible to calcium deregulation, with resultant cell death via calcium mediated pathways. Therefore, improving NCX activity following cerebral ischaemia may provide a therapeutic strategy to reduce neuronal death.

Parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHRP) regulate Na$ sp+$/H$ sp+$ exchanger (NHE) activity in various types of cells such as osteoblastic cells and renal proximal tubule OK cells. Na$ sp+$/H$ sp+$ exchangers are plasma membrane transporters catalyzing the electroneutral exchange of 1 H$ sp+$ for 1 Na$ sp+.$ Several mammalian isoforms of NHE have been so far identified with each mediating a variety of specific functions. Parathyroid hormone, playing an essential role in the physiology of blood Ca$ sp{2+}$ and phosphate homeostasis, inhibits renal proximal tubular bicarbonate reabsorption by inhibiting the apically located Na$ sp+$/H$ sp+$ exchanger. However, which specific isoform of NHE mediated this effect and the specific signaling components involved were unknown. In our studies we determined that Na$ sp+$/H$ sp+$ exchanger NHE-3 isoform is expressed in the renal proximal tubule OK cells and that N-terminal PTH and PTHRP analogues upon binding to their receptor stimulate both the PKA and the PKC pathways, each of which can independently lead to inhibition of this exchanger activity. NHEs also play an important role in the regulation of intracellular pH which is subject to fluctuation occurring during the process of hormone stimulated bone formation and bone remodeling. Again the specific NHE isoform(s) mediating this effect and the signaling pathways involved were unidentified. It is determined by our studies that NHE type 1 is expressed in osteoblastic cell line, UMR-106 cells, and that PTH and PTHRP stimulate this exchanger via a cAMP-dependent pathway exclusively. It was believed that motivation of NHE-1 in the UMR-106 cells and inhibition of NHE-3 in the OK cells by N-terminal analogues of PTH and PTHRP involves binding of these analogues to a common G protein-coupled receptor called the "classical" PTH/PTHRP receptor. However, with the recent discovery of other PTH and/or PTHRP receptor, this hypothesis is no longer clear

Tidal exchanges of organic carbon, nitrogen and phosphorus between a south temperate Sarcocornia marsh and its associated estuary are examined. Subterranean water flow was small, and the hydraulic exchange between the two systems largely surficial. The dominant tidal signal was semi-diurnal, and the extent of inundation of the marsh varied considerably as a consequence of interactions of semi-lunar tidal cycles with changes in daily mean sea level. Annual net fluxes of organic carbon were directed from the marsh to the estuary, but amounted to less than 2% of marsh aerial net primary productivity. This indicates the incompatibility of E.P. Odum's outwelling hypothesis to this marsh-estuarine system. The direction of net flux of organic carbon switched on a time-scale of days. These directions were largely correlated with mesoscale oceanic events, which materially altered the extent of marsh inundation, and which provided evidence of the mutual exclusivity of outwelling of DOC from the marsh and oceanic upwelling. Laboratory mesocosm experiments using intact marsh blocks of sediment from the marsh were conducted to identify the proximate processes and interactions at the marsh-water interface responsible for the variability of marsh-estuarine exchanges. Patterns of fluxes of organic carbon, total nitrogen and phosphorus were markedly different in the structurally contrasted tidal creek and Sarcocornia Zone regions of the marsh. Both regions exported these components, but the fluxes of organic carbon and total phosphorus were significantly larger from the tidal creek than from the Sarcocornia zone, and the opposite applied to nitrogen. The presence of brachyuran crabs . the most numerous macrofauna on the marsh enhanced the flux of carbon, nitrogen and phosphorus from the marsh biocoenosis, largely as a result of the effect of their bioturbation. Evidence is examined which suggests that differential mobilization of nutrients in the two zones by crabs is responsible for biogeochemical coupling of these two regions , which may account for the elevated productivity of salt- marsh systems

Trypanosoma cruzi est un parasite protozoaire à multiplication intracellulaire, agent de la maladie de Chagas, infectant 16 à 18 millions de personnes en Amérique latine. Il peut être transmis de la mère infectée au fœtus dans 2 à 10 % des cas, mais ses autres effets sur la gestation ont été peu étudiés. Par ailleurs, les cytokines ont des effets sur la gestation. Certaines d’entre elles, comme l’interleukine-1, l’IL-4, l’IL-5, l’IL-10, le GM-CSF et le TGF-b2, sont bénéfiques pour la gestation, tandis que d’autres, comme l’IL-2, l’IL-12, l’IFN-g et le TNF-a ont des effets nocifs sur celle-ci. L’impact de l’infection à T. cruzi, stimulant la production de TNF-a et d’IFN-g, sur l'implantation et la croissance fœtale n’a pas été étudié.

Le but de notre travail était d’étudier les effets de l’infection aiguë à T. cruzi sur la capacité de reproduction de la souris. Nous avons ainsi évalué les effets de cette infection sur la fertilité, le développement et la viabilité des fœtus de souris et le rôle de l’IFN-g et du TNF produits au cours de l’infection sur le développement de la gestation.

Nous avons montré que l’infection aiguë à T. cruzi :i) diminue la capacité de reproduction de la souris ;ii) provoque une mortalité fœtale massive précoce (résorptions), tardive et néonatale associée à un retard de croissance intra-utérin, et ce, iii) en dehors de toute transmission congénitale du parasite.

Par ailleurs nos travaux montrent que la mortalité fœtale/néonatale est associée à une invasion parasitaire massive du placenta qui présente d’importantes lésions à type d’infiltrats inflammatoires, de nécrose ischémique, de dépôts de fibrine et de thromboses vasculaires. Nous avons noté qu’il existe une relation inverse entre la charge parasitaire des unités utéro-placentaires et la viabilité du conceptus, suggérant que ces lésions placentaires contribuent à la mortalité fœtale en limitant les échanges materno-fœtaux.

Enfin, nous avons également étudié le rôle de cytokines abortogènes comme le TNF et l’IFN-g, produites abondamment pendant l’infection aiguë de la souris par T. cruzi. Les taux sanguins maternels d’IFN-g étaient augmentés au 9ième mais pas aux 17ième et 19ième jours de gestation, alors que les taux de TNF sanguin et la production placentaire de cette cytokine augmentaient aux 17ième et 19ième jours de gestation. Afin d’évaluer le rôle de ces deux cytokines dans la mortalité fœtale, des souris ont été traitées par la pentoxifylline, pour inhiber la transcription du gène de TNF-a et diminuer la production d’IFN-g. Ces souris montraient une réduction de la mortalité fœtale à mi-gestation, associée à une diminution de la production du TNF placentaire, sans modifications des taux systémiques et sans effets sur l’IFN-g, suggérant la contribution du TNF dans la mortalité fœtale associée à l’infection aiguë par T. cruzi.

En conclusion, notre travail montre que l’infection aiguë à Trypanosoma cruzi exerce un effet particulièrement néfaste sur la capacité de reproduction et le développement de la gestation chez la souris et que les lésions placentaires liées à l’infection et la production de TNF par le placenta infecté contribuent à cet effet.

Doctorat en sciences biomédicales
info:eu-repo/semantics/nonPublished

O$\sb2$ transport was examined by measuring the fractional saturation of concentrated hemoglobin solutions flowing through an artificial capillary (diameter $\approx$ 27 $\mu$m). The measured effects of pH, hemoglobin concentration, O$\sb2$ tension, temperature, and organic phosphate were analyzed by a mathematical model which included the geometry of the capillary, parabolic flow inside the lumen, and cooperative O$\sb2$ binding. Oxygen exchange was limited by diffusion and therefore governed by the magnitude of the O$\sb2$ gradient between the intracapillary fluid phase and the external gas space. In uptake experiments, O$\sb2$ flux was determined primarily by the external O$\sb2$ tension (160 mm Hg) because the internal O$\sb2$ pressure was kept small due to chemical combination with hemoglobin. In release experiments, the external O$\sb2$ tension was maintained at zero, and the transport rate was determined by the intracapillary oxygen partial pressure, which was proportional to the O$\sb2$ half-saturation pressure of the hemoglobin sample. Thus, factors that change the affinity of hemoglobin for oxygen, such as pH, temperature, and organic phosphate concentration, influence strongly the rate of O$\sb2$ release but have little effect on the rate of O$\sb2$ uptake. The rate-limiting step for erythrocyte CO$\sb2$ transport is the transmembrane exchange of Cl$\sp-$ and HCO$\sb3\sp-$ anions. This process was measured by following extracellular pH changes using a pH-sensitive fluorescent dye in a stopped-flow mixing device equipped with front-face optics. Initial pH and chloride gradients induced pH relaxations which were sensitive to the specific inhibitor 4,4$\prime$-diisothiocyano-2,2$\prime$-stilbenedisulfonic acid (DIDS). Special mixing experiments, designed to minimize anion competition for the transporter binding site(s), were simulated mathematically by models written for ping-pong and random ternary complex mechanisms. The experimentally observed rate dependence on initial extracellular bicarbonate concentration was approximated better by the ping-pong model, and the theoretically derived values for Cl$\sp-$ and HCO$\sb3\sp-$ binding (both K$\sb {\rm D}$'s = 5 mM) as well as the translocation rate (1.1 $\times$ 10$\sp4$ s$\sp{-1}$) agreed very well with literature values for inhibition constants and protein turnover number, respectively. Finally the simultaneous measurement of O$\sb2$ uptake and subsequent HCO$\sb3\sp-$/Cl$\sp-$ exchange was demonstrated. The hemoglobin color change upon oxygen binding caused a rapid change in fluorescence, followed by a slower, DIDS-sensitive pH equilibration.

The goal of this work is to study human whole-body gas exchange and cerebral autoregulation using a mathematical model. Previously, a human cardiopulmonary (CP) model [45, 47] was developed, which included heart, closed-loop blood circulation, gas exchange at lungs and baroreflex control of arterial pressure. In the current study, two major extensions to the model are made. First, a description of gas exchange in the peripheral tissues is added and is coupled with the lung gas exchanger via the circulatory loop with variable transport delays. A peripheral chemosensitive loop is also added to mimic the influence of blood gas composition on the heart and vasculature. The CP model is then used to predict the integrated cardiovascular and blood-tissue gas transport responses to pronounced changes in lung gas composition, and thus simulates changes encountered in apnea with and without passive oxygenation. The second extension of the CP model includes a more detailed description of cerebral circulation, cerebrospinal fluid (CSF) dynamics, brain gas exchange and cerebral blood flow (CBF) autoregulation. Two CBF regulatory mechanisms are described: autoregulation and CO2 reactivity. Central chemoreceptor control of ventilation is also added. This new model is subsequently used to study cerebral hemodynamic and brain gas exchange responses to test protocols commonly used in the assessment of CBF autoregulation (e.g., carotid artery compression and the thigh cuff test). The model closely mimics the experimental findings and provides biophysically based insights into the dynamics and interactions of the associated physiological systems. In summary, this work represents a bold effort in large-scale modeling of physiological systems. The presented model accurately describes the physiological systems and can explain how the cardiovascular, pulmonary and autonomic nervous systems interact in response to a variety of cardiopulmonary challenges, such as apnea, carotid artery compression and the thigh cuff test. With further refinement, the model may help investigators to better understand the complex biophysics of cardiopulmonary diseases such as sleep-related disorders of breathing (obstructive and central sleep apnea) and complications associated with head-injuries.

碩士
國立臺南藝術大學
藝術史與藝術評論研究所
93
This paper mainly is the method that learns through Archeology to discourse the evolution of the metal animal-shaped plates on northern of China before Han Dynasty. Focuses on the evolution of form and styles of the metal animal-shaped plates itself, and also to discourse the cultural exchange between northern and central plains region of China. First chapter includes motivation, purpose, the review of history and methods of this study. Second chapter describes archeological distribution of the animal-shaped plates on northern of China before Eastern Han Dynasty, showing the growth or the declination of the two types among the four regions on China’s northern. Third chapter focuses on the special features of the four regions on China’ northern. Fourth chapter concludes the interaction between function and form by way of the funerary condition of the metal animal-shaped plates, this chapter also to discourse the cultural exchange between northern and central plains region of China. Fifth chapter concern the conclusion and the lack of this thesis.

Further in vivo functional studies were also performed. The SLC26A3 antibody was injected into the BALB/C mice seminiferous tubules using micropipette. The animals were sacrificed after three days, and CASA, daily sperm production (DSP) were used to evaluate sperm motility and spermatogenesis. The results showed that sperm motility was increased while there was no significant difference between DSP. Our results indicate that SLC26A3 on sperm does not play a dominant role in spermatogenesis, epididymal maturation and sperm motility.
In the first part of study, guinea pig sperm which were incubated in medium with various concentrations of Cl- resulted in varied percentages of capacitated sperm, in a concentration dependent manner. Depleting Cl-, even in the presence of HCO3 -, abolished sperm capacitation and vice versa, indicating the involvement of both anions in the process. Capacitation-associated HCO 3- dependent events, including cAMP production, protein tyrosine phosphorylation and pHi increase also depend on Cl - concentrations. Similar Cl- dependence was observed for sperm hyperactivated motility and sperm-egg fusion. The capacitation-associated events could also be significantly reduced by inhibitors or antibodies of CFTR and SLC26A3, with a more potent effect observed for niflumate, an inhibitor more selective for SLC26A3, over that of DIDS, an inhibitor more selective for SLC4 exchangers. The expression and localization of CFTR and SLC26A3 in guinea pig sperm were also demonstrated using immunostaining and Western blot analysis. Our results indicate that Cl- is required for the entry of HCO3- necessary for sperm capacitation, implicating the involvement of SLC26A3 in transporting HCO3 - with CFTR providing the recycling pathway for Cl- .
In the second part of study, GC-1 spg cell line that expresses SLC26A6 but not SLC26A3 was used as a negative control. The cells and sperm were pretreated with anion exchanger inhibitors and SLC26A3 antibody, and then membrane potential and intracellular calcium were measured. Our results showed that DIDS could inhibit the HCO3- deficiency induced depolarization of GC-1 spg cells as well as the depolarization induced by Cl- or HCO3- deficiency in sperm. Niflumate could inhibit the HCO3- induced [Ca 2+] i increase of the sperm but not GC-1 spg cells. SLC26A3 antibody had no effect on the GC-1 spg cells but it could block the depolarization caused by C--deficiency in sperm.
Our previous study has demonstrated the involvement of Cystic fibrosis transmembrane conductance regulator (CFTR) in transporting bicarbonate necessary for sperm capacitation. However, whether its involvement is direct or indirect remains unclear. The present study is design to investigate: (1) the possibility of a Cl-/HCO3- exchanger, solute carrier family 26, number 3 (SLC26A3), operating with CFTR during sperm capacitation, (2) the role and the underlying mechanisms of SLC26A3 in other sperm post-testicular processes and spermatogenesis.
Taken together, our results demonstrate the involvement of SLC26A3 in sperm function, particularly in transporting HCO3- necessary for sperm capacitation, which appears to be working with CFTR providing the recycling pathway for Cl- in parallel. The present results also provide an explanation to the observed subfertility in patients with SLC26A3 mutations. Further in vitro and in vivo studies also have shown that SLC26A3 does not play a predominant role in spermatogenesis but may affect other post-testicular maturation processes.
Chen, Wenying.
"November 2009."
Adviser: H.C. Chan.
Source: Dissertation Abstracts International, Volume: 72-01, Section: B, page: .
Thesis (Ph.D.)--Chinese University of Hong Kong, 2010.
Includes bibliographical references (leaves 101-109).
Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web.
Abstract also in Chinese.