Orientador: Lizandra Amoroso / Resumo: A urolitíase é enfermidade com importância econômica para a ovinocultura, que causa a saída prematura de ovinos machos destinados à reprodução, custos com tratamento e morte. A associação entre avaliação hemogasométrica, pesquisa de proteínas de fase aguda (PFA) como biomarcadores precoces e o estudo morfométrico do trato urinário pode ser empregada como método de auxílio diagnóstico precoce para a doença. Com esse objetivo, foram utilizados 14 ovinos hígidos, machos (não castrados), da raça Santa Inês com idade aproximada de 90 dias. Os ovinos receberam dieta experimental hiperfosfórica durante todo o período experimental, foram examinados semanalmente e após desenvolvimento da urolitíase, reorganizados em dois grupos experimentais distintos D1 (sem urolitíase) e D2 (com urolitíase) para análise comparada dos dados. No período pré-experimental e no dia do abate foram coletadas amostras de sangue venoso para avaliação hemogasométrica. Para mensurar as imunoglobulinas (A e G) e as PFA, analisaram-se as amostras dos ovinos que desenvolveram a urolitíase (D2). As coletas de sangue foram realizadas semanalmente até a manifestação clínica da enfermidade, totalizando 16 amostras. Ao término do experimento foi realizado o abate e necropsia dos ovinos, para descrição das alterações patológicas e a análise morfométrica. Fragmentos do trato urinário e fígado foram coletados e submetidos à rotina histológica e as lâminas histológicas foram descritas, seguindo-se a histomorfometria. Embo... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Urolithiasis is an economically important disease for male sheep production that causes the premature calving of sheep for breeding, treatment costs and death. The association between hemogasometric evaluation, acute phase protein (APP) research as early biomarkers and urinary tract morphometric study can be employed as an early diagnostic aid for the disease. For this purpose, 14 healthy male (non-castrated) Santa Inês sheep, approximately 90 days old, were used. Sheep received a hyperphosphoric experimental diet throughout the experimental period, were examined weekly and after development of urolithiasis, reorganized into two distinct experimental groups D1 (without urolithiasis) and D2 (with urolithiasis) for comparative analysis of the data. In the pre-experimental period and on the day of slaughter, venous blood samples were collected for hemogasometric evaluation. To measure immunoglobulins (A and G) and APP, samples from sheep that developed urolithiasis (D2) were analyzed. Blood samples were collected weekly until the clinical manifestation of the disease, totaling 16 samples. At the end of the experiment, sheep were slaughtered and necropsied to describe the pathological changes and the morphometric analysis. Urinary tract and liver fragments were collected and submitted to histological routine and histological slides were described, followed by histomorphometry. Although blood pH was not different (P < 0.05) between groups, sheep that developed urolithiasis had com... (Complete abstract click electronic access below) / Doutor

Photophysical properties of an array of various polyaromatic hydrocarbons were benchmarked with B3LYP, M06 and B97D methods coupled with Pople and CEP-31G(d) basis sets. Results from the benchmark show the importance of diffuse basis sets when modeling the electronic properties of highly conjugated systems and provide qualitative reliable accuracy with certain levels of theory. B97D and M06 are applied to modeling pyrene adducts governed by non-covalent interactions in both gaseous and condensed states to reproduce experimental spectra. DFT calculations with both B97D and M06 functionals show qualitatively and quantitatively that pyrene dimer is a stronger π–base as compared to its monomer. Binding energies coupled with MEP, PCA and Qzz results show that the difference in π-basicity of the monomer and dimer impacts the supramolecular chemistry involved in adducts formed with super π-acidic silver cyclometallic trimer (CTC). Non-covalent interactions between coinage metal CTCs and ammonia/phosphine substrates is reported. Interactions between these substrates and the facial plane of the π-rich gold CTC reveal a novel interaction, where the typical Lewis acid/base roles are reversed for the substrates. Adducts formed through this type of interaction define typical Lewis bases like ammonia and phosphine as Lewis acids, wherein the partially positive hydrogens coordinate to the metallo-aromatic center through dipole-quadrupole interactions. Interactions of ammonia at the side positions is shown to heavily impact the Lewis basicity of the CTC facial plane leading to similar interactions exhibited by the ammonia-gold CTC adducts. Structural and electronic properties of the adducts modeled are examined.

The possibility that an inadequate response of the pulmonary system might limit high intensity exercise in man has received increasing attention over the past few years. However, very few scientific investigations have focused systematically on pulmonary function during high intensity constant-load exercise. Furthermore, many studies have examined only one part of the pulmonary system during exercise and some have not included blood gas measurements as a measure of the adequacy of pulmonary function. The studies reported in this thesis were designed to investigate the possible failure of the gas exchanging and pump functions of the pulmonary system during high intensity constant-load exercise. In particular, the aim was to determine the extent to which the pulmonary system might be a factor causing fatigue during this form of exercise.

This study examined the acid-base and ionoregulatory responses by the skin and kidney of adult bullfrogs (Rana catesbiana) during recovery from non-respiratory acidoses. Acidosis was induced by intravascular infusions of HCl (3,000 uequiv/kg), or NH4Cl (4,000 uequiv/kg), or by 45 min enforced submergence. Infused acids were immediately buffered by the extracellular fluids (ECF) and moved rapidly into the intracellular compartment. Clearance of the acid load was slow (> 24 h post-infusion) and only in NH4Cl loaded frogs was the full amount cleared within 5 days. Excess acid was excreted primarily by increased renal NH4+ effluxes. The skin contributed very little to the net acid excretion; instead large "base" losses ("acid" uptake) occurred. Acid infusions also resulted in large ion losses and elevated water uptake across the skin, with electrolytes and water moving down chemical gradients. Frogs infused with HCl died within 96 h; NH4Cl loaded frogs all survived.Forced submergence resulted in a severe lactacidosis which was corrected in 12 h by a combination of renal/cutaneous acid excretion (9:1 ratio) and metabolic utilization of lactate and H+ (~ 95% of excess acid load). Acid excretion occurred primarily as NH4+ efflux by the kidney and skin. A 1:1 exchange of Na+ influx/NH4+, efflux across the skin was found (r = 0.94; P < 0.01) when Jin Na+ was elevated over basal levels (30-40 uequiv/kg·h). Lactate and net acid effluxes by the skin and kidney diminished by 24 h post-disturbance and accounted for 4 to 6% of the total estimated load to the animal. All frogs survived the diving protocol. Restoration of acid-base state in acidotic bullfrogs by ECF and non-ECF mechanisms, as well as the renal and cutaneous responses to the acidosis, is discussed and compared to that of other vertebrates. It is concluded that anurans, and the bullfrog in particular, are poor regulators of acid-base state but instead are very tolerant to marked disturbances of acid-base balance. / Thesis / Master of Science (MS)

Azidotische Belastungen des Pansens gelten bei Milchkühen besonders in der Frühlaktation als weit verbreitet. Sie werden als subakute Pansenazidosen (SARA) für die häufigen Produktionskrankheiten verantwortlich gemacht. Ziele : dieser Untersuchungen waren a) die Kontrolle des Säure-Basen-Haushalt (SBH) bei HF- Hochleistungskühen im peripartalen Zeitraum zum Vorkommen von SARA, b) die Prüfung bewährter sowie neuer Methoden zur Pansensaftanalyse, c) die Analyse von SBH-Harnparametern sowie d) die Prüfung möglicher Beziehungen von Stoffwechselparametern im Blut zu SBH-Parametern im Pansensaft. Versuchsanordnung : In 10 Betrieben Niedersachsens wurden an je 10 Kühen Verlaufsuntersuchungen mit vier Proben/Tier durchgeführt: 1. Trockenstehperiode/Färsen -30 Tage ante partum (d a.p.) bis zur Kalbung; 2. 2 bis 7 Tage post partum (d p.p.); 3. in der Frühlaktation 8 bis 30 d p.p. und 4. in der Mittellaktation 80 bis 150 d p.p. Pansensaft vormittags mit Geishauser- Sonde gewonnen, Blut aus der V. jugularis externa sowie Harn per Blasenkathederisierung. Analysiert wurden im Pansensaft der pH-Wert, die Pufferkapazität als Titrationsbilanz über die Titrationsazidität und - alkalität, die Methylenblauprobe sowie die Sedimentaktivitätszeit (SAT), im Harn der pH-Wert, die Netto-Säure-Basen-Ausscheidung (NSBA) sowie der Basen-Säure-Quotient (BSQ) und im Blutserum Glukose, freie Fettsäuren (FFS), Betahydroxybutyrat (BHB), L-Laktat, Cholesterol, Protein, Albumin, Bilirubin, Harnstoff, Kreatinin, Na, K, Cl, Ca, P, Mg, ALP, ASAT, GLDH sowie die CK. Ergebnisse: Zwischen Kühen und Färsen wurden bei den gemessenen Parametern keine gesicherten Unterschiede festgestellt. Es gab auch keine Unterschiede zwischen den Betrieben. Die Pansen-pH-Werte bewegten sich von x= 6,8 (6,43/7,0;1./3. Quartil) zu Beginn des Trockenstehens über minimal x = 6,3 (5,9/6,7) 30 bis 60 d p.p. bis x = 6,6 (6,2/6,8) in der Mittellaktation. Sie lagen alle im physiologischen Bereich. 20 Kühe hatten Pansen-pH-Werte von < 5,8, 6 Kühe von < 5,5 und 21 Kühe von > 7,2. Damit kamen die Kühe der SARA-Definition, bezogen auf das einmalige Auftreten von pH-Werten < 5,8 bei einer Kuh, nahe; bezogen auf alle gemessenen Pansen-pH-Werte betrug der Anteil < 5,8 aber nur 9,8 %. Die Titrationsbilanz war in der Trockensteherperiode ausgeglichen; bei allen drei Kontrollen p.p. bestand ein Basendefizit. Die Pufferkapazität war in der 1. Woche p.p. und in der Frühlaktation vermindert. Panse-pH-Werte und Titrationsbilanz korrelierten sehr eng (r=0,98,p<0,001). Die Methylenblauprobe war a.p. mit x = 3:37 (2:22/4:59) Minuten am längsten, in der Mittellaktation mit 3:01 (2:25/4:30) Minuten am kürzesten und ingesamt physiologisch. Es gab keine signifikanten Unterschiede zwischen den einzelnen Kontollzeiträumen. Die SAT war physiologisch und schwankte zwischen zwei und fünf Minuten ohne signifikante Unterschiede zwischen den Entnahmezeiträumen. Im Harn lagen die NSBA mit x = 70 mmol/l (20/122 = 1./3.Quartil) und der BSQ mit x = 2,2 (1,6/2,8) nur bei den Frischmelkern unter den unteren Grenzwerten und deuteten die Gefahr von SARA an. Der Harn-pH-Wert war mit x = 7,98 (7,7/8,2) physiologisch. Von der Gesamtheit alle NSBA-Werte waren 37% < 83 mmol/l sowie 28,9% < 25 mmol/l; beim BSQ waren es 52,6% < 2,5 sowie 27,4% < 1,5. Die für die NSBA, den BSQ und den Harn-pH-Wert berechneten Sensivitäten und Spezifitäten für einen Pansen-pH-Wert < 5,8 liegen mit 44,8% und 64,5% (NSBA), 55,8% und 47,9% (BSQ) sowie 2,3% und 97% (Harn-pH-Wert) unter den diagnostischen Erfordernissen.Sie können somit keine sicheren Rückschlüsse auf den Pansenzustand geben. Bei den Blutbefunden wurden Kühe mit Pansen-pH-Werten < 5,8 (SARA) und >5,8 gegenüber gestellt. Die Korrelationskoeffizienten, die Sensitivität und die mittels Receiver-Operating-Characteristics Analysen ermittelten Flächen unter der Kurve zwischen Pansen-pH-Wert und den Blutparametern waren niedrig ( p> 0,05). Schlussfolgerung: Hochleistende HF-Kühe haben peripartal nicht zwangsläufig eine SARA. Die Titrationsbilanz ergänzt den Pansensaft-pH-Wert, ist routinemäßig aber nicht nötig. Im Harn liegen die NSBA und BSQ häufiger unter den Grenzwerten und korrelieren nicht gesichert mit dem Pansen-pH-Wert. Auch Blutparameter erlauben keine sichere Aussage über den Pansenzustand. Die Diagnose von SARA muss mit direkter Messung des Pansen-pH-Wertes gestellt werden.

Azidotische Belastungen des Pansens gelten bei Milchkühen besonders in der Frühlaktation als weit verbreitet. Sie werden als subakute Pansenazidosen (SARA) für die häufigen Produktionskrankheiten verantwortlich gemacht. Ziele : dieser Untersuchungen waren a) die Kontrolle des Säure-Basen-Haushalt (SBH) bei HF- Hochleistungskühen im peripartalen Zeitraum zum Vorkommen von SARA, b) die Prüfung bewährter sowie neuer Methoden zur Pansensaftanalyse, c) die Analyse von SBH-Harnparametern sowie d) die Prüfung möglicher Beziehungen von Stoffwechselparametern im Blut zu SBH-Parametern im Pansensaft. Versuchsanordnung : In 10 Betrieben Niedersachsens wurden an je 10 Kühen Verlaufsuntersuchungen mit vier Proben/Tier durchgeführt: 1. Trockenstehperiode/Färsen -30 Tage ante partum (d a.p.) bis zur Kalbung; 2. 2 bis 7 Tage post partum (d p.p.); 3. in der Frühlaktation 8 bis 30 d p.p. und 4. in der Mittellaktation 80 bis 150 d p.p. Pansensaft vormittags mit Geishauser- Sonde gewonnen, Blut aus der V. jugularis externa sowie Harn per Blasenkathederisierung. Analysiert wurden im Pansensaft der pH-Wert, die Pufferkapazität als Titrationsbilanz über die Titrationsazidität und - alkalität, die Methylenblauprobe sowie die Sedimentaktivitätszeit (SAT), im Harn der pH-Wert, die Netto-Säure-Basen-Ausscheidung (NSBA) sowie der Basen-Säure-Quotient (BSQ) und im Blutserum Glukose, freie Fettsäuren (FFS), Betahydroxybutyrat (BHB), L-Laktat, Cholesterol, Protein, Albumin, Bilirubin, Harnstoff, Kreatinin, Na, K, Cl, Ca, P, Mg, ALP, ASAT, GLDH sowie die CK. Ergebnisse: Zwischen Kühen und Färsen wurden bei den gemessenen Parametern keine gesicherten Unterschiede festgestellt. Es gab auch keine Unterschiede zwischen den Betrieben. Die Pansen-pH-Werte bewegten sich von x= 6,8 (6,43/7,0;1./3. Quartil) zu Beginn des Trockenstehens über minimal x = 6,3 (5,9/6,7) 30 bis 60 d p.p. bis x = 6,6 (6,2/6,8) in der Mittellaktation. Sie lagen alle im physiologischen Bereich. 20 Kühe hatten Pansen-pH-Werte von < 5,8, 6 Kühe von < 5,5 und 21 Kühe von > 7,2. Damit kamen die Kühe der SARA-Definition, bezogen auf das einmalige Auftreten von pH-Werten < 5,8 bei einer Kuh, nahe; bezogen auf alle gemessenen Pansen-pH-Werte betrug der Anteil < 5,8 aber nur 9,8 %. Die Titrationsbilanz war in der Trockensteherperiode ausgeglichen; bei allen drei Kontrollen p.p. bestand ein Basendefizit. Die Pufferkapazität war in der 1. Woche p.p. und in der Frühlaktation vermindert. Panse-pH-Werte und Titrationsbilanz korrelierten sehr eng (r=0,98,p<0,001). Die Methylenblauprobe war a.p. mit x = 3:37 (2:22/4:59) Minuten am längsten, in der Mittellaktation mit 3:01 (2:25/4:30) Minuten am kürzesten und ingesamt physiologisch. Es gab keine signifikanten Unterschiede zwischen den einzelnen Kontollzeiträumen. Die SAT war physiologisch und schwankte zwischen zwei und fünf Minuten ohne signifikante Unterschiede zwischen den Entnahmezeiträumen. Im Harn lagen die NSBA mit x = 70 mmol/l (20/122 = 1./3.Quartil) und der BSQ mit x = 2,2 (1,6/2,8) nur bei den Frischmelkern unter den unteren Grenzwerten und deuteten die Gefahr von SARA an. Der Harn-pH-Wert war mit x = 7,98 (7,7/8,2) physiologisch. Von der Gesamtheit alle NSBA-Werte waren 37% < 83 mmol/l sowie 28,9% < 25 mmol/l; beim BSQ waren es 52,6% < 2,5 sowie 27,4% < 1,5. Die für die NSBA, den BSQ und den Harn-pH-Wert berechneten Sensivitäten und Spezifitäten für einen Pansen-pH-Wert < 5,8 liegen mit 44,8% und 64,5% (NSBA), 55,8% und 47,9% (BSQ) sowie 2,3% und 97% (Harn-pH-Wert) unter den diagnostischen Erfordernissen.Sie können somit keine sicheren Rückschlüsse auf den Pansenzustand geben. Bei den Blutbefunden wurden Kühe mit Pansen-pH-Werten < 5,8 (SARA) und >5,8 gegenüber gestellt. Die Korrelationskoeffizienten, die Sensitivität und die mittels Receiver-Operating-Characteristics Analysen ermittelten Flächen unter der Kurve zwischen Pansen-pH-Wert und den Blutparametern waren niedrig ( p> 0,05). Schlussfolgerung: Hochleistende HF-Kühe haben peripartal nicht zwangsläufig eine SARA. Die Titrationsbilanz ergänzt den Pansensaft-pH-Wert, ist routinemäßig aber nicht nötig. Im Harn liegen die NSBA und BSQ häufiger unter den Grenzwerten und korrelieren nicht gesichert mit dem Pansen-pH-Wert. Auch Blutparameter erlauben keine sichere Aussage über den Pansenzustand. Die Diagnose von SARA muss mit direkter Messung des Pansen-pH-Wertes gestellt werden.

Acid-base equilibria in organic particulate matter (PM) are poorly understood, but have important implications for air quality and public health. First, acid-base reactions in organic particulate matter affect the gas/particle partitioning of organic compounds in the atmosphere, and these processes are not currently represented in atmospheric and climate change models. Second, the acid-base balance of tobacco smoke affects the amount of nicotine absorbed by the smoker, and a greater understanding of this balance would help to relate cigarette smoke composition to the addictive properties of cigarettes. This work presents data related to both air quality and tobacco smoke modeling. The gas/particle partitioning behavior of organic acids and bases is highly dependent on acid-base equilibria and speciation between neutral and ionic forms, because ionic compounds do not volatilize. Descriptions of acid dissociation behavior in atmospheric PM have, to date, focused primarily on phases in which the solvent is water; however, atmospheric PM may include up to 90% organic matter. Data is presented here describing the acid dissociation behavior of organic acids and protonated amines in organic/aqueous mixtures (chosen to approximate the characteristics of organic PM) with varying levels of water content. In such mixtures, the preferential solvation of ions and neutral molecules (by the aqueous portion or the organic portion, respectively) affects the acid-base equilibria of the solutes. It is demonstrated that neutralization reactions between acids and bases that create ions are likely to have non-negligible effects on gas/particle partitioning under certain atmospheric conditions. Thus, including acid-base reactions in organic gas/particle partitioning models could result in a greater proportion of acidic and basic compounds partitioning to the particulate phase. In addition, the acid dissociation constants (pKa values) of atmospherically-relevant acids and bases vary with water content. Specifically, as water content increases, the pKa values of organic acids decrease dramatically, while the pKa values of protonated amines changes only slightly. This situation can result in drastically different speciations and partitioning behavior depending on water content. This second part of this work reports some of the data needed to develop an acid-base balance for tobacco smoke PM using electroneutrality as a governing principle. Five brands of cigarettes were sampled and the smoke PM extracted. Cations (sodium, potassium, and ammonia) and anions (organic acids, nitrate, nitrite, and chloride) were measured using ion chromatography. Ammonia and organic acids were also re-measured after the acidification of the sample in order to determine whether "bound" forms of these compounds exist in cigarette PM. Weak acids were determined by acid-base titration to determine whether or not all of the weak acids (including organic acids) had been accounted for by the ion chromatography. Weak bases were also determined by acid-base titration, and the majority of weak base is expected to be accounted for by total nicotine (to be measured in a separate analysis). In terms of total acidic species and total basic species, two of the five cigarette brands measured were relatively basic, and three were relatively acidic. Between 50% and 89% of the titrated acids were accounted for by the anionic species measured in ion chromatography. Based on samples tested after sample acidification, about half of the potential ammonia in tobacco smoke PM exists in "bound" form. The speciation of weak acids and bases in tobacco smoke PM cannot be determined from this data alone, because the equilibrium constants of acid-base reactions are not understood in complex organic media. The data presented here, when combined with data from free-base and total nicotine analyses, represent a first step toward a predictive model of acid-base behavior in tobacco smoke PM.

The current increase in the atmospheric CO2 concentration results in two major consequences in the marine environment: an increase of the sea surface temperature (0.7 °C since pre-industrial times) and a decreased seawater pH. This decrease is being measured continuously in different parts of the world and ranges from -0.0017 to -0.04 units per year according to the location considered. Based on CO2 emissions models provided by the IPCC, it was predicted that the average open ocean pH would decrease further by 0.4 units by 2100 and 0.8 by 2300 (corresponding to about a three-fold and six-fold increase of the proton concentration). Also, saturation states of seawater for the different forms of calcium carbonate, such as calcite, magnesium calcite and aragonite which are produced by calcifying marine organisms, are decreasing and consequently the saturation horizons of these minerals are shoaling. Today, some environments are characterized by pH values lower than the average open ocean pH. These are intertidal rock pools, upwelling zones, the deep-sea and CO2 vents. In these environments, pH is either constantly low or fluctuates. Those changes are either due to biological activity, geological CO2 leakage, or water masses movements. Within these environments, it has been hypothesized that organisms could be adapted or acclimatized to low pH values such as those predicted for the near-future. <p><p>Tolerance to ocean acidification in metazoans is linked to their acid-base regulation capacities when facing environmental hypercapnia (i.e. increased CO2 concentration in the surrounding environment). The latter may result in a hypercapnia of the internal fluids and a concomitant acidosis (i.e. reduced pH of the internal fluids due to the dissociation of CO2 in this case). Organisms have two buffer systems allowing the compensation of this acidosis: the CO2-bicarbonate and the non-bicarbonate buffers. Homeostasis of the internal fluids thanks to these systems is essential for the proper functioning of enzymes and processes. As hypometabolic calcifying osmoconformers, three of the characteristics conferring a relative vulnerability to ocean acidification, echinoderms are considered “at risk” for the near-future conditions. Nonetheless, post-metamorphic (juveniles and adults) echinoderms inhabit all environments showing naturally low pH. Furthermore, sea urchins which are highly calcified (compared to sea stars or sea cucumbers) are also found in these environments. This suggests that echinoderms have strategies to adapt or acclimate to low pH environments. Recent studies indicated that while sea urchins are able to regulate their coelomic (extracellular) fluid by accumulation of bicarbonate, sea stars seem to tolerate the acidosis linked to environmental hypercapnia. However, this information was obtained on a reduced number of species and significant interspecific differences were evidenced. Some taxa have not been investigated at all. Furthermore, several aspects of the acid-base physiology were unexplored, like the buffering capacity of the extracellular fluid and the origin of carbon within these fluids.<p><p>Accordingly, the goal of this study was to characterize the acid-base physiology in post-metamorphic echinoderms of different taxa in order to understand their response to ocean acidification.<p><p>The acid-base regulation capacities within the different echinoderm taxa were compared. A method was designed to measure the total alkalinity in small volumes (500 µl) of the main extracellular fluid (the coelomic fluid). This study showed that regular euechinoids have an increased buffer capacity in their coelomic fluid compared to seawater and the other echinoderm groups. In sea urchins, bicarbonate and non-bicarbonate buffers come into play, the former playing the major role. This buffer capacity was increased in fed individuals compared to fasted ones and increased further when seawater pH was lowered.<p><p>The acid-base regulation capacities of sea urchins from different taxa were investigated. Regular euechinoids possess an increased buffer capacity of the coelomic fluid allowing them to maintain a higher pH compared to cidaroids at current seawater pH. This pattern was found for temperate, tropical and Antarctic sea urchins. Data was also obtained for irregular echinoids which also showed a particularly low extracellular pH and a buffer capacity close to seawater like cidaroids. When exposed to reduced seawater pH (8.0, 7.7, and 7.4) for 4-6 weeks, regular euechinoids showed an increasing buffer capacity of the coelomic fluid accompanied by a homeostasis of the pH. On the contrary, cidaroids showed no changes in their acid-base status whatever the seawater pH (8.0 to 7.4). The origin of coelomic fluid carbon, investigated by stable carbon isotope analysis, also differs according to taxa. The δ13CDIC of regular euechinoids evidenced a mixing between CO2 from metabolic origin and that from the surrounding seawater. This is further supported by the correlation between the seawater signal of reduced pH conditions (modified by the addition of industrial gas, changing the δ13C to more negative values) and that of the coelomic fluid. On the other hand, cidaroids exhibit a signal reflecting principally metabolic CO2 (very negative δ13C), and the δ13C did not change under varying pH conditions (i.e. did not adapt to the seawater δ13CDIC signature). For irregular echinoids, the carbon origin is unclear as some species show signals close to that of regular euechinoids whereas others are similar to cidaroids.<p><p>The impact of acid-base regulation was investigated by testing the effect of ocean acidification on the mechanical properties of the skeleton (test plates) in the sea urchin Paracentrotus lividus. Individuals from intertidal pools, CO2 vents and a one year acidification experiment (pH 8.0, 7.9 and 7.7) were compared. Only the intertidal pool individuals showed a difference of the Young’s modulus and fracture forces of their plates. Sea urchins from the tide pool with the largest pH fluctuations showed a lower stiffness and strengthened test. On the contrary, sea urchins from CO2 vents and experimental acidification did not display any differences in the several mechanical properties tested. We suggest that the different food qualities (calcified vs. uncalcified algae) in the different tide pools significantly contributed to the observed difference.<p><p>The acid-base regulation ability of sea cucumbers was assessed in two species from contrasted habitats (mangrove intertidal vs. coral reef species). These organisms underwent acidosis of the coelomic fluid when exposed to reduced seawater pH for a short time (6 to 12 days). The δ13C signal of the coelomic fluid mirrored that of the surrounding seawater in all conditions, indicating that the CO2 accumulated (cause of the acidosis) comes also from the seawater. This is still unexplained to date. However, metabolic processes such as respiration and ammonium excretion rates were not affected. No difference was evidenced between the two species.<p><p>The results obtained in this study compiled with data from the literature indicate that post-metamorphic echinoderms have contrasted acid-base physiology with most regular euechinoids compensating the coelomic fluid pH by accumulation of bicarbonate ions (and possibly ophiuroids also), cidaroids and at least one regular euechinoid (Arbacia lixula) having a naturally low coelomic fluid pH which is not affected by acidification, and sea stars and sea cucumbers which do not compensate their coelomic fluid pH when submitted to acidified seawater. In regular euechinoids, negative effects are linked to resource allocation with growth usually being reduced in favor of acid-base regulation mechanisms. Starfish and sea cucumbers appear as resilient to acidification, with very few functions being negatively impacted. In conclusion, it seems that post-metamorphic echinoderms studied so far will not be particularly at risk when facing ocean acidification levels expected by 2100. Furthermore, tolerance to ocean acidification does not seem linked to the present day ambient pH regime. Nevertheless, more studies need to be carried out on brittle stars and sea cucumbers to confirm preliminary results, as well as crinoids which have not been investigated to date. Long-term exposure experiments to estimate energy budget changes as well as more assessments of evolutionary potential in echinoderms are crucially needed./L’augmentation actuelle de la concentration en CO2 atmosphérique a deux conséquences majeures dans l’environnement marin :une augmentation de la température des eaux de surface (0.7°C depuis l’époque préindustrielle) et une diminution du pH de l’eau de mer. Cette diminution est mesurée continuellement dans différentes régions du monde et varie de -0.0017 à -0.04 unités de pH par an en fonction du site considéré. Basé sur des modèles d’émissions de CO2 du GIEC, il a été prédit que le pH moyen de l’océan diminuerait encore de 0.4 unités d’ici 2100 et 0.8 d’ici 2300 (correspondant à une augmentation de la concentration en protons d’environ 3 fois et 6 fois). De même, les états de saturation de l’eau de mer vis-à-vis des différentes formes de carbonate de calcium, telles que la calcite, la calcite magnésienne et l’aragonite produites par les organismes calcifiants, sont en train de diminuer et par conséquent, les horizons de saturation remontent vers les eaux de surface. Aujourd’hui, certains environnements sont caractérisés par des valeurs de pH plus basses que celle de l’océan. Ceux-ci sont les mares intertidales, les zones d’upwelling, l’océan profond et les évents volcaniques. Dans ces environnements, le pH est soit constamment bas ou fluctue. Ces changements sont soit dû à une activité biologique, une fuite de CO2 géologique, ou au mouvement des masses d’eau. Dans ces environnements, il a été suggéré que les organismes pourraient être adaptés ou acclimatés à des valeurs basses de pH, telles que celles prédites pour le futur proche.<p> <p>La tolérance à l’acidification des océans chez les métazoaires est liée à leur capacité de régulation acide-base lorsqu’ils sont exposés à une hypercapnie environnementale (c’est-à-dire, une augmentation de la concentration en CO2 dans l’environnement entourant l’organisme). Ce phénomène peut résulter en une hypercapnie des liquides internes et une acidose concomitante (c’est-à-dire, un pH des liquides internes réduit dû à la dissociation du CO2 dans ce cas précis). Les organismes ont deux systèmes tampons leur permettant de compenser l’acidose :les tampons CO2-bicarbonate et non-bicarbonate. L’homéostasie des liquides internes grâce à ces systèmes est essentielle pour le fonctionnement correct des enzymes et processus. En tant qu’osmoconformes calcifiant hypométaboliques, trois caractéristiques menant à une certaine vulnérabilité face à l’acidification des océans, les échinodermes sont considérés « à risque » pour les conditions du futur proche. Cependant, les échinodermes post-métamorphiques (juvéniles et adultes) occupent tous les environnements montrant un pH faible naturellement. De plus, les oursins qui sont hautement calcifiés (par rapport aux étoiles de mer ou aux concombres de mer) sont également retrouvés dans ces environnements. Ceci suggère que les échinodermes ont des stratégies d’adaptation ou d’acclimatation à ces environnements à bas pH. Alors que des études récentes montrent que les oursins sont capables de réguler le pH du liquide cœlomique (extracellulaire) par l’accumulation de bicarbonates, les étoiles semblent tolérer l’acidose liée à l’hypercapnie environnementale. Néanmoins, ces informations ont été obtenues sur un petit nombre d’espèces et des différences interspécifiques significatives ont été mises en évidence. Certains taxa n’ont pas été étudié du tout. Par ailleurs, différents aspects de la physiologie acide-base sont inexplorés, tels que la capacité tampon du liquide extracellulaire et l’origine du carbone dans ces liquides.<p><p>Par conséquent, le but de cette étude était de caractériser la physiologie acide-base chez les échinodermes post-métamorphiques de différents taxa afin de comprendre leur réponse à l’acidification des océans.<p><p>Les capacités de régulation acide-base au sein des différents groupes d’échinodermes ont été comparées. Une méthode a été mise au point afin de mesurer l’alcalinité totale dans de petits volumes (500 µl) de liquide extracellulaire (le liquide cœlomique). Cette étude démontra que la capacité tampon du liquide cœlomique des euéchinoïdes réguliers est accrue comparée à celle de l’eau de mer ainsi que celle des autres groupes d’échinodermes. Dans les oursins, les tampons bicarbonate et non-bicarbonate entrent en jeux, le premier étant majoritaire. Cette capacité tampon est augmentée chez les individus nourris par rapport à ceux à jeuns et est augmentée plus encore lorsque le pH de l’eau de mer est diminué.<p><p>Les capacités de régulation acide-base ont été étudiées plus spécifiquement dans les différents groupes d’oursins. Les euéchinoïdes réguliers possèdent une capacité tampon accrue du liquide cœlomique leur permettant de maintenir un pH élevé comparé aux oursins cidaroïdes, au pH de l’eau de mer actuel. Ce patron se retrouve dans les oursins tempérés, tropicaux et antarctiques. Des données ont également été obtenues pour les oursins irréguliers qui ont également un pH extracellulaire particulièrement bas et une capacité tampon proche de celle de l’eau de mer comme les cidaroïdes. Lorsqu’ils sont exposés à un pH de l’eau de mer réduit (7.7 et 7.4 par rapport à 8.0) pour 4 à 6 semaines, les euéchinoïdes réguliers ont montré une augmentation de la capacité tampon du liquide cœlomique accompagnée d’une homéostasie du pH de ce liquide. A l’inverse, les cidaroïdes n’ont montré aucune modification de leur statut acide-base quel que soit le pH (8.0 à 7.4). L’origine du carbone du liquide cœlomique, étudié par analyse des isotopes stables du carbone, diffère également en fonction du groupe. Le δ13CDIC des euéchinoïdes réguliers met en évidence un mélange entre du CO2 d’origine métabolique et celui de l’eau environnante. Ceci est également démontré par la corrélation entre le signal de l’eau de mer dont le pH est réduit (modifié par l’ajout de CO2 industriel, changent le δ13C vers des valeurs plus négatives) et celui du liquide cœlomique. En revanche, les cidaroïdes montrent un signal reflétant principalement celui du CO2 métabolique (δ13C très négatif), et le δ13C n’est pas influencé par des conditions de pH variées (c’est-à-dire, qu’il ne s’adapte pas à la signature du δ13CDIC de l’eau de mer). Pour les oursins irréguliers, l’origine du carbone est incertaine puisque certaines espèces montrent un signal proche de celui des euéchinoïdes réguliers et d’autres similaire à celui des cidaroïdes.<p><p>L’impact de la régulation acide-base a été étudié en testant l’effet de l’acidification des océans sur les propriétés mécaniques du squelette (plaques squelettiques) de l’oursin Paracentrotus lividus. Des individus de mares intertidales, d’évents volcaniques et d’une expérience d’acidification d’un an (pH 8.0, 7.9 et 7.7) ont été comparés. Seuls les individus des mares intertidales montrèrent une différence pour le module de Young et la force des fractures des plaques. Les oursins venant de la mare intertidale montrant les plus grandes variations de pH avaient une rigidité plus faible et un squelette renforcé. A l’inverse, les oursins des évents volcaniques et de l’expérience d’acidification n’ont montrés aucune différence dans les diverses propriétés mécaniques étudiées. Nous suggérons que les variations en termes de qualité de nourriture (algues calcifiées vs. non-calcifiées) dans les différentes mares intertidales ont contribués de manière significative à la différence observée.<p><p>L’habilité des concombres de mer à réguler leur balance acide-base a été évaluée dans deux espèces d’habitats contrastés (espèce intertidale des mangroves vs. subtidale des récifs coralliens). Ces organismes ont subis une acidose du liquide cœlomique lorsqu’ils ont été exposés à un pH réduit de l’eau de mer pour une courte durée (6 à 12 jours). Le signal δ13C du liquide cœlomique reflétait celui de l’eau environnante dans toutes les conditions, indiquant que le CO2 accumulé (cause de l’acidose) venait de l’eau. Ceci est encore inexpliqué à l’heure actuelle. Cependant, les processus métaboliques tels que la respiration ou l’excrétion d’ammonium n’ont pas été affecté. Aucune différence n’a été observée entre les deux espèces.<p><p>Les résultats obtenus dans cette étude compilés avec ceux de la littérature indiquent que les échinodermes post-métamorphiques ont une physiologie acide-base contrastée avec la plupart des euéchinoïdes réguliers qui compensent le pH du liquide cœlomique par l’accumulation d’ions bicarbonates (et peut-être les ophiures aussi), les cidaroïdes et au moins un euéchinoïde régulier (Arbacia lixula) qui ont naturellement un pH du liquide cœlomique bas et qui ne sont pas affectés par l’acidification, et les étoiles de mer et les concombres de mers qui ne compensent pas le pH du liquide cœlomique lorsqu’ils sont soumis à une eau acidifiée. Chez les euéchinoïdes réguliers, des effets négatifs sont liés à un changement de l’allocation des ressources avec souvent un taux de croissance réduit en faveur des mécanismes de régulation acide-base. Les étoiles de mer et les concombres de mer apparaissent plus tolérants à l’acidification, avec peu de fonctions négativement impactées. En conclusion, il semble que les échinodermes post-métamorphiques étudiés jusqu’à présent ne seront pas particulièrement à risque lorsqu’ils seront exposés au niveau d’acidification attendu pour 2100. De plus, la tolérance à l’acidification des océans ne semble pas liée au régime de pH subit actuellement. Cependant, plus d’études doivent être menées sur les ophiures et les concombres de mer afin de confirmer les résultats préliminaires, ainsi que sur les crinoïdes qui n’ont à l’heure actuelle pas encore été étudiés. Des expériences à long terme afin d’estimer le budget énergétique des organismes ainsi que plus d’évaluations du potentiel d’évolution chez les échinodermes sont absolument nécessaires.<p> / Doctorat en Sciences / info:eu-repo/semantics/nonPublished

Thesis ((DMed)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: We have developed an innovative multimedia e-learning resource, the Electrolyte Workshop, to provide students and clinicians with instruction and the opportunity for simulated practice in managing electrolyte and acid-base disorders. Our teaching approach is built around relevant physiology and makes use of real cases and storytelling to engage the learner. We have documented the challenges encountered during the development process and have made recommendations for the managing of similar projects. While there are many factors that must be in place to ensure successful e-learning, this dissertation focuses on an important but under-appreciated factor, namely the usability of the computer interface. Usability describes how easy technology interfaces are to use and is routinely evaluated and optimized in the software development industry. This is not yet the case with e-learning, especially in the area of medical education. Poor usability limits the potential benefit of educational resources, as this means that learners will struggle with the interface as well as with the challenges of the content presented. A comprehensive usability evaluation of our Electrolyte Workshop was completed. This included testing with typical end-users, where data were collected via standardized questionnaires and by observing and analysing their interactions with our application. We employed heuristic evaluation as an additional approach and assembled a panel of experts to evaluate our application against a set of heuristics, or principles of good interface design. Many serious usability problems were identified, thus severely limiting the potential educational impact of our Electrolyte Workshop. There was a striking disconnect between the objective measures of usability and self-reported questionnaire data. Our user-testing data make a useful contribution to the debate on how many users are required to find most of the usability problems in an interface. Heuristic evaluation proved to be a very efficient approach. However, both user testing and heuristic evaluation detected serious problems which were missed with the other method. These evaluations informed a comprehensive revision of our application and we could then compare the original with an optimized version in a randomized trial. We found large improvements in objective usability measures, which are likely to increase the satisfaction and motivation of learners. There were similar scores on measures of learning. This was not surprising as our participants were all relatively high-knowledge learners and not novices as regards the subject matter. Our study clearly indicates that the usability evaluation of e-learning resources is critical, and provides an example of how clinician-teachers can improve the usability of the resources they develop. Usability should be evaluated as a routine part of the development and implementation of e-learning materials, modules and programmes. This should start with the earliest versions of the resource, when making changes is easier and less costly. We have demonstrated that a combination of methods should be employed and have highlighted the utility of heuristic evaluation. An iterative approach should be followed, with several cycles of testing and re-design. User testing should always include the study of objective usability measures and not rely only on self-reported measures of user satisfaction. / AFRIKAANSE OPSOMMING: Ons het ’n innoverende multimediahulpbron vir e-leer, die Electrolyte Workshop, ontwikkel om studente en klinici van ’n onderrighulpmiddel sowel as die geleentheid vir gesimuleerde oefening in die hantering van elektroliet en suur-basis stoornisse te voorsien. Ons onderrigbenadering is gegrond op relevante fisiologie en maak gebruik van werklike gevalle en vertelkuns om die leerder te betrek en te boei. Ons het die uitdagings gedurende die ontwikkelingsproses opgeteken en aanbevelings oor die bestuur van soortgelyke projekte gedoen. Hoewel suksesvolle e-leer van etlike faktore afhang, konsentreer hierdie verhandeling op ’n belangrike dog onderskatte faktor, naamlik die bruikbaarheid van die rekenaarkoppelvlak. Bruikbaarheid verwys na die gemak waarmee tegnologiekoppelvlakke gebruik kan word, en word gereeld in die sagtewareontwikkelingsbedryf beoordeel en verbeter. Tog is dit nog nie die geval met e-leer nie, veral op die gebied van mediese onderrig. Swak bruikbaarheid beperk die moontlike voordeel van opvoedkundige hulpbronne, aangesien leerders voor die dubbele uitdaging van ’n ingewikkelde koppelvlak én die voorgeskrewe inhoud te staan kom. ’n Omvattende bruikbaarheidsbeoordeling is van die Electrolyte Workshop onderneem. Dit het toetsing met tipiese eindgebruikers ingesluit, waarvoor data met behulp van gestandaardiseerde vraelyste ingesamel en gebruikers se interaksie met die toepassing waargeneem en ontleed is. Ons het heuristiese evaluering as bykomende benadering gebruik en ’n kennerspaneel saamgestel om ons toepassing aan die hand van ’n stel heuristiek, oftewel beginsels van goeie koppelvlakontwerp, te beoordeel. ’n Hele aantal ernstige bruikbaarheidsprobleme is uitgewys, wat die moontlike opvoedkundige impak van die Electrolyte Workshop erg beperk. Daar was merkbare teenstrydigheid tussen die objektiewe bruikbaarheidsmaatstawwe en die selfaangemelde vraelysdata. Ons gebruikerstoetsdata lewer ’n waardevolle bydrae tot die debat oor hoeveel gebruikers nodig is om die meeste van die bruikbaarheidsprobleme met ’n koppelvlak te ontdek. Heuristiese evaluering was ʼn baie doeltreffende benadering. Tog het gebruikerstoetsing op sekere ernstige probleme afgekom wat heuristiese evaluering misgekyk het, en andersom. Hierdie beoordelings het as grondslag gedien vir ’n omvattende hersiening van die toepassing, waarna ons die oorspronklike weergawe in ’n verewekansigde proef met ’n verbeterde weergawe kon vergelyk. Die objektiewe bruikbaarheidsmaatstawwe het groot verbeterings getoon, wat waarskynlik leerders se tevredenheid en motivering sal verhoog. Leermaatstawwe het soortgelyke tellings opgelewer. Dít was egter te wagte gewees, aangesien die deelnemers almal betreklik ingelig was oor die vakmateriaal, eerder as nuwelinge. Ons studie het bevestig dat die bruikbaarheidsbeoordeling van e-leerhulpbronne noodsaaklik is, en bied ’n voorbeeld van hoe klinici-opvoeders bruikbaarder hulpbronne kan ontwikkel. Bruikbaarheid behoort as ’n roetinedeel van die ontwikkeling en inwerkingstelling van e-leermateriaal, -modules en -programme beoordeel te word. Dit behoort reeds by die vroegste weergawes van die hulpbron te begin, wanneer dit makliker en goedkoper is om veranderinge aan te bring. Ons het ook getoon dat ’n kombinasie van metodes gebruik behoort te word, en het die nut van heuristiese evaluering beklemtoon. ’n Herhalende benadering moet gevolg word, met etlike siklusse van toetsing en herontwerp. Gebruikerstoetsing behoort altyd die beoordeling van objektiewe bruikbaarheidsmaatstawwe in te sluit, en moenie slegs op selfaangemelde maatstawwe van gebruikerstevredenheid staatmaak nie.

Betanina e indicaxantina foram obtidas e purificadas e a hidrólise térmica de betanina foi investigada em diferentes pHs, concentrações de tampão, temperaturas e com a adição de sais. Para a decomposição de indicaxantina, foi realizado um estudo em diferentes pHs. Tampões acetato e citrato se mostraram adequados para estudos com betanina, dentro de sua faixa de tamponamento. Os experimentos realizados com tampão fosfato indicaram a ocorrência de catálise geral pelo íon fosfato presente no tampão. O efeito dos sais sobre a hidrólise de betanina foi investigado em pHs 3 e 6,2, com a adição de sais de sódio, amônio e sais cloretos. p-Toluenosulfonato de sódio, cloreto de amônio e cloreto tetrabutilamônio foram os sais que apresentaram maior efeito sobre a constante cinética observada de hidrólise de betanina. A hidrólise das duas betalaínas apresentou um comportamento semelhante com a variação do pH e o ajuste multiparamétrico dos dados sugere que a catálise básica específica é a principal responsável pela hidrólise de betanina e indicaxantina. Um modelo que sugere a abertura do anel 1,2,3,4-tetrahidropiridínico das betalaínas é proposto para explicar o efeito do pH sobre a constante cinética de decomposição determinada em meio ácido. / Betanin and indicaxanthin were obtained and purified and the thermal hydrolysis of betanin was investigated at different pH, buffer concentration, temperature and in the presence of different salts. For the decomposition of indicaxanthin, a study was conducted at different pHs. Acetate buffer and citrate buffer proved to be suitable for the study of betanin. The experiments carried out with phosphate buffer indicated the occurrence of general base catalysis by phosphate ion present in the buffer. The effect of salts on the betanin hydrolysis was investigated at pH 3 and 6.2 with the addition of sodium salts, ammonium salts and chlorides. Sodium p-toluenesulfonate, ammonium chloride and tetrabutylammonium chloride had the larger effects on the observed rate constant for the hydrolysis of betanin. The variation of pH resulted in similar hydrolysis profile for both betalains studied and multiparameter fitting of the data suggests that the specific base catalysis is primarily responsible for the hydrolysis of betanin and indicaxanthin. A model that suggests the opening of the 1,2,3,4-tetrahydropyridine ring of betalains is proposed to explain the effect of pH on the decomposition rate constant under acidic conditions.