El síndrome ascítico es una de las principales causas de mortalidad en aves de producción de carne, siendo la hipoxia el factor desencadenante a la que se suma la respuesta del organismo mediante la liberación de diversas sustancias que conducen al cuadro patológico, tales como endotelina 1 (ET-1), óxido nítrico (ON), prostaglandinas y diversas citoquinas inflamatorias. Aún no se conoce el rol del óxido nítrico en la hipertrofia arteriolar pulmonar e hipertrofia cardiaca derecha que se observa en el síndrome ascítico. Por lo tanto, el objetivo del presente estudio fue determinar los valores de nitritos y nitratos, metabolitos estables del ON y su correlación con el grado de hipertrofia arteriolar pulmonar mediante la relación capa muscular/diámetro arteriolar (CM/DA) y con la hipertrofia cardiaca derecha mediante la relación ventrículo derecho/ventrículo total (VD/VT) y relación ventrículo derecho/peso corporal (VD/PC) en aves sometidas a hipoxia ambiental. De un mismo lote de aves machos de la línea Cobb vantres, nacidos a nivel del mar, se estudiaron 135 animales; 15 fueron sacrificados a 1 día de edad, en sus muestras se estudiaron diversas variables cuyos resultados fueron la base comparativa para los que se obtuvieron en los demás animales. Los 120 animales restantes fueron divididos en los grupos: nivel del mar (NM) 60 animales y altura (A) 60 animales. A los 10, 20, 30 y 40 días de edad se tomaron 15 aves (al azar) de cada grupo.Las variables determinadas fueron peso corporal (PC), hematocrito (Ht), nitritos y nitratos, CM/DA, VD/VT y VD/PC. / Ascitic syndrome is one of the main causes of mortality in broilers, being the hypoxia the leading factor to which is added the answer of the organism by liberation of diverse sustances that conduce to the pathologic pattern, such us endotheline 1 (ET-1), nitric oxide (NO), prostaglandins and inflammatory chemokines. The rol of NO in the pulmonary arteriolar hypertrophy and right cardiac hypertrophy observed in ascitic syndrome is still unknown. Hence, the aim of this study was to obtain the values of nitrites and nitrates, stable metabolites of NO, and its correlation with the degree of pulmonary arteriole hypertrophy through the muscular wall/arteriolar diameter (CM/DA) ratio, the right ventricular hypertrophy through the right ventricle/total ventricle (RV/TV) ratio and right ventricle/body weight (RV/BW) ratio in chickens rised at environmental hypoxia. A total of 135 male, Cobb vantres chickens and born at sea level were studied; 15 of them were sacrificed at 1 day age and diverse variables were determinate in their samples whose results were used as comparative base line for those of the other animals. The reminder 120 chickens were distributed in the groups: Sea Level (SL) 60 birds and altitude (A) 60 birds. At 10, 20, 30 and 40 days old, 15 chicken were taken randomly to measure body weight (BW), hematocrite (Ht), nitrites and nitrates, MW/AD, RV/VT and RV/BW.

Some aspects of anuran metabolism are examined, with special emphasis on possible limitations to aerobic metabolism and the effect of chronic hypoxia acclimation on maximal rates of aerobic metabolism and the metabolic fate of lactic acid accumulated after anaerobic metabolism.

Master of Science (Medicine) / Clinical studies have demonstrated an arousal deficit in infants suffering Obstructive Sleep Apnoea (OSA), and that treatment to alleviate the symptoms of OSA appears to reverse the deficit in arousability. Some sudden infant deaths are thought to be contingent upon such an arousal deficit. This research utilised young piglets during early postnatal development, and exposed them to intermittent hypercapnic hypoxia (IHH) as a model of clinical respiratory diseases. Arousal responses of control animals were compared to the animals exposed to IHH. Comparisons were also made between successive exposures on the first and the fourth consecutive days of IHH. Time to arouse after the onset of the respiratory stimulus, and frequency of arousals during recovery, demonstrated that arousal deficits arose after successive exposures and that these were further exacerbated on the fourth study day. After an overnight recovery period, the arousal deficit was apparently dormant, and only triggered by HH exposure. These studies confirm that both acute and chronic deficits can be induced on a background of otherwise normal postnatal development, suggesting that deficits observed in the clinical setting may be a secondary phenomenon.

Perinatal hypoxia-ischaemia is a major cause of disability, including cerebral palsy, yet a neuroprotectant which fully protects the brain remains elusive. Following a hypoxic-ischaemic insult, striatal medium-spiny neurons and hippocampal CA1 neurons are vulnerable to a complex cascade of neurotoxic events. This cascade includes energy failure, a massive release of glutamate, the formation of free radicals and caspase activation. The overall aim of this thesis was to assess the efficacy of three potential neuroprotective strategies that target this cascade from different directions. Short-term, and where appropriate, long-term, neuroprotection was investigated. The first treatment strategy aimed to suppress the generation of free radicals through treatment with the potent free radical spin trap, N-tertbutyl-(2-sulphophenyl)-nitrone (S-PBN). The second compound tested was the caspase-3 inhibitor, minocycline. Finally, the third treatment strategy combined a series of S-PBN injections with 6 hours of moderate hypothermia immediately after hypoxia-ischaemia. Hypothermia is suggested to slow the rate of the neurotoxic cascade, thus potentially allowing other neuroprotective agents greater efficacy. Using an adaptation of the Rice et al. (1981) model, hypoxia-ischaemia was induced on postnatal day (PN) 8 in the right cerebral hemisphere. For the short-term studies, the rats were perfused at 14 days-of-age. The brains were dissected out and embedded in Technovit. Forty [mu]m serial sections were cut through the right striatum and hippocampus. The total number of medium-spiny neurons in the striatum and where appropriate, the total number of neurons in the hippocampal CA1 pyramidal layer, were stereologically determined using the optical disector/Cavalieri method. For the long-term study, fine motor control was assessed in half of the animals through the staircase test from 9-11 weeks-of-age. Neuroprotection was assessed in the remaining animals. All animals were sacrificed at 12 weeks-of-age. The total number of striatal medium-spiny neurons was stereologically determined in the non-behavioural animals as described above. A series of seven injections of S-PBN (100mg/kg) did not offer statistically significant neuroprotection to the striatum at one week after perinatal hypoxia-ischaemia. Similarly, a single injection of minocycline (45mg/kg) immediately after the insult did not offer significant neuroprotection to the striatum nor the CA1 region of the hippocampus at this early time-point. In contrast, when the series of S-PBN injections was combined with 6 hours of moderate hypothermia post-hypoxia-ischaemia, sterelogical analysis revealed significant neuroprotection of the striatal medium-spiny neurons to normal levels at one week after the injury. No significant neuroprotection was seen in the CA1 region of the same animals. To assess whether this impressive striatal neuroprotection was long-lasting and whether it represented functional rescue, the final experiment in this thesis investigated rat pups at 12 weeks-of-age after exposure to hypoxia-ischaemia at PN8. Treatment with S-PBN/hypothermia offered persistent neuroprotection of striatal medium-spiny neurons and preservation of fine motor skills compared to diluent-normothermia-treated controls. The long-term behavioural outcomes were compared with normal, uninjured controls and the total number of medium-spiny neurons was compared with normal numbers from the literature. These comparisons revealed that the histological and functional integrity of the striatum was rescued to normal levels. This is the first study to identify a treatment strategy that offers complete and long-lasting preservation of striatal neuronal numbers, by accurate and unbiased stereological methods, paired with persistent preservation of fine motor control following perinatal hypoxia-ischaemia.

Subsequent to an hypoxic-ischaemic (HI)-insult a multi-faceted complex cascade of events occurs that ultimately results in cellular and neurological impairments within cortical and sub-cortical central nervous system (CNS) regions. In the present studies a modified �Levine� rat-pup model of HI (left carotid artery ligation + 1 hour global hypoxia on post-natal day (PND) 26) was employed to assess the neuroprotective properties of clomethiazole (CMZ; a γ-aminobutyric acid (GABA)A receptor agonist). In this study, histological and electrophysiological paradigms were used to assess the long-term neuroprotective properties of CMZ (414mg/kg/day via mini-pumps). Key enzymes involved in inflammation, namely nitric oxide synthase (NOS) and arginase, were also examined to assess other potential CMZ mechanisms. Assessments were carried out 3- and 90-days post-HI, with extensive ipsilateral CNS lesions evident at a gross histological level, at both the early and long-term stages, with CMZ significantly decreasing the lesion size at 3- and 90-days (P<0.01; P<0.05). Evoked field potential analyses were used to assess hippocampal CA1 neuronal activity ex vivo. Electrophysiological measurements contralateral to the occlusion revealed impaired neuronal function following HI relative to short- and long-term controls (P<0.001, 3- and 14-days; P<0.01, 90-days), with CMZ providing near complete protection (P<0.001 at 3- and 14-days; P<0.01 at 90-days). Both inducible NOS (iNOS) and arginase activities were significantly increased at 3-days (P<0.01), with arginase activity remaining elevated at 90-days post-HI (P<0.05) ipsilaterally. CMZ suppressed the HI-induced increase in iNOS and arginase activities (P<0.001; P<0.05). These data provide evidence of long-term functional neuroprotection afforded by CMZ in a model of HI-induced neurodegeneration. In addition, under conditions of HI, functional deficits were not restricted to the ipsilateral hemisphere and were due, at least in part, to changes in the activity of NOS and arginase. Underlying mitochondrial dysfunction is eminently present in many neuropathological conditions. The full extent of mitochondrial dysfunction in cortical, hippocampal and cerebellar tissues was assessed following HI. Assessment of mitochondrial FAD-linked respiration at both 1- and 3-days post-HI revealed a significant decrease in activity from ipsilateral cortical and hippocampal regions (P<0.001). In addition, significant changes in respiratory function were also evident in contralateral regions and cerebellum, 3-days post-HI (P<0.05). Assessment of the mitochondrial electron transport chain (complexes I-V) and mitochondrial markers of integrity (citrate synthase) and oxidative stress (aconitase) confirmed ipsilateral mitochondrial impairment following HI. Complexes I, II-III, V and citrate synthase were also impaired, in contralateral regions and cerebellum, 3-days post-HI. CMZ treatment provided significant protection to all mitochondrial aspects of neuronal tissue assessed. This study provides evidence of the full extent of mitochondrial damage following an HI-insult and may contribute, in part, to the impairment seen contralaterally. In addition, protection afforded by CMZ extends to preservation of mitochondrial function and integrity. Cerebral ischaemia-induced angiogenesis has been shown within and around infarcted regions and may contribute to a more favourable neurological outcome. The level of angiogenesis was examined using platelet endothelial cell adhesion molecule-1 (PECAM-1 / CD31). CD31 immunolabelling 7-days post-HI revealed a significant increase in angiogenesis compared with non-intervention controls (P<0.001). Treatment with CMZ decreased the level of angiogenesis compared to HI + saline (P<0.001) back to non-intervention control levels. Conversely, N[omega]-nitro-L-arginine methyl ester (L-NAME) treatment (5mg/kg/day) exacerbated the ischaemic lesion (P<0.001) and resulted in a marked decrease in angiogenesis compared to non-intervention controls (P<0.001). The extent of cerebral infarction in these studies is dependent on the level of NOS activity with CMZ increasing total NOS levels compared to HI + saline, while L-NAME halted the HI-induce increase in total NOS activity (P<0.001). These results show for the first time, that angiogenesis may be used as an assessment of neurodegeneration / neuroprotection in pathologies of cerebral ischaemia and are directly correlated with changes in NOS activity. These studies have therefore shown that following HI, damage also occurs contralateral to the occlusion, and is not restricted to the ipsilateral hemisphere. In addition, the neuroprotective effects of CMZ have been shown to extend out to 90-days post-HI, whereby significant protection to CA1 neuronal activity was seen. These studies also provide in vivo evidence that CMZ may also afford neuroprotection via anti-inflammatory pathways, as evidenced by a decrease in iNOS and arginase activities. Furthermore, these studies have also show evidence that angiogenesis (CD31) can be used as a diagnostic tool to assess neuroprotection / neurodegeneration.

Hypoxia is a common feature of many respiratory disorders including acute severe asthma, chronic obstructive pulmonary disease and pneumonia. Hypoxia also occurs during sleep - disordered breathing in conditions such as sleep hypoventilation syndrome and sleep apnea. In most respiratory diseases hypoxia is coupled with increased respiratory load. Compensatory protective mechanisms are activated to oppose these impediments to respiration. However, hypoxia is associated with impaired neurocognitive function and recent studies have demonstrated that hypoxia suppresses respiratory load perception in healthy individuals and asthma patients. These recent findings raise the possibility that a variety of protective physiological reflex responses to increased respiratory load may be impaired during periods of hypoxia. The effects of hypoxia on several of these protective responses and possible mechanisms of respiratory sensory depression by hypoxia are explored in the experiments outlined in this thesis. In the first study, the respiratory related evoked potential ( RREP ) was used to investigate the mechanisms underlying hypoxia - induced suppression of respiratory load sensation in healthy individuals. As a positive control the effects of hypoxia on respiratory load perception to inspiratory resistive loads were also measured. The amplitude of the first and second positive peaks ( P1 and P2 ) of the RREP were significantly reduced during hypoxia. P1 is thought to reflect the arrival of the ascending respiratory signals to the somatosensory area of the cortex. The perceived magnitude of externally applied inspiratory resistive loads was also reduced during hypoxia. These data provide further support that hypoxia suppresses respiratory load perception and suggest that this is mediated, at least in part, by suppression of respiratory afferent information prior to its arrival at the cortex. In the second study, the effects of acute sustained hypoxia on the cough reflex threshold and cough tachyphylaxis to inhaled capsaicin were explored in healthy individuals. Acute sustained hypoxia suppressed cough reflex sensitivity to inhaled capsaicin. This finding raises the possibility that the cough reflex, important for protecting the lungs from inhalation or aspiration of potentially injurious substances and for clearing excess secretions, may be impaired during acute exacerbations of hypoxic - respiratory disease. In the third study, reflex responses of the genioglossus and scalene muscles to brief pulses of negative airway pressure were compared between hypoxia and normoxia during wake and sleep in healthy males in the supine position. Cortical RREPs to the same stimuli were also examined under these conditions. The genioglossus is the largest upper airway ( UA ) dilator muscle and can be reflexively augmented in response to negative UA pressure. A diminished response of this muscle during sleep has been postulated to be a contributing mechanism to obstructive sleep apnea ( OSA ) in individuals with an anatomically narrow UA. Cortical activation ( i.e. arousal ) to sudden airway narrowing in OSA is an important protective response to help restore ventilation during an obstructive event. In this study, genioglossus reflex responses to negative pressure pulse stimuli were maintained during mild overnight hypoxia. Conversely, reflex inhibition of the scalene muscle to the same stimuli was prolonged during hypoxia. In addition, a previously undescribed morphology of the genioglossus negative pressure reflex consisting of activation followed by suppression was observed with greater suppression during sleep than wake. The amplitude of the P2 component of the RREP was also significantly reduced during hypoxia. In summary, the potential mechanisms underlying hypoxia - induced suppression of respiratory load sensation and the effects of hypoxia on several protective respiratory responses have been investigated in healthy subjects. The potential implications of these findings for patients with hypoxic - respiratory disease are discussed. / Thesis (Ph.D.)--School of Molecular and Biomedical Science, 2006.

El síndrome ascítico es una de las principales causas de mortalidad en aves de producción de carne, siendo la hipoxia el factor desencadenante a la que se suma la respuesta del organismo mediante la liberación de diversas sustancias que conducen al cuadro patológico, tales como endotelina 1 (ET-1), óxido nítrico (ON), prostaglandinas y diversas citoquinas inflamatorias. Aún no se conoce el rol del óxido nítrico en la hipertrofia arteriolar pulmonar e hipertrofia cardiaca derecha que se observa en el síndrome ascítico. Por lo tanto, el objetivo del presente estudio fue determinar los valores de nitritos y nitratos, metabolitos estables del ON y su correlación con el grado de hipertrofia arteriolar pulmonar mediante la relación capa muscular/diámetro arteriolar (CM/DA) y con la hipertrofia cardiaca derecha mediante la relación ventrículo derecho/ventrículo total (VD/VT) y relación ventrículo derecho/peso corporal (VD/PC) en aves sometidas a hipoxia ambiental. De un mismo lote de aves machos de la línea Cobb vantres, nacidos a nivel del mar, se estudiaron 135 animales; 15 fueron sacrificados a 1 día de edad, en sus muestras se estudiaron diversas variables cuyos resultados fueron la base comparativa para los que se obtuvieron en los demás animales. Los 120 animales restantes fueron divididos en los grupos: nivel del mar (NM) 60 animales y altura (A) 60 animales. A los 10, 20, 30 y 40 días de edad se tomaron 15 aves (al azar) de cada grupo.Las variables determinadas fueron peso corporal (PC), hematocrito (Ht), nitritos y nitratos, CM/DA, VD/VT y VD/PC. / Ascitic syndrome is one of the main causes of mortality in broilers, being the hypoxia the leading factor to which is added the answer of the organism by liberation of diverse sustances that conduce to the pathologic pattern, such us endotheline 1 (ET-1), nitric oxide (NO), prostaglandins and inflammatory chemokines. The rol of NO in the pulmonary arteriolar hypertrophy and right cardiac hypertrophy observed in ascitic syndrome is still unknown. Hence, the aim of this study was to obtain the values of nitrites and nitrates, stable metabolites of NO, and its correlation with the degree of pulmonary arteriole hypertrophy through the muscular wall/arteriolar diameter (CM/DA) ratio, the right ventricular hypertrophy through the right ventricle/total ventricle (RV/TV) ratio and right ventricle/body weight (RV/BW) ratio in chickens rised at environmental hypoxia. A total of 135 male, Cobb vantres chickens and born at sea level were studied; 15 of them were sacrificed at 1 day age and diverse variables were determinate in their samples whose results were used as comparative base line for those of the other animals. The reminder 120 chickens were distributed in the groups: Sea Level (SL) 60 birds and altitude (A) 60 birds. At 10, 20, 30 and 40 days old, 15 chicken were taken randomly to measure body weight (BW), hematocrite (Ht), nitrites and nitrates, MW/AD, RV/VT and RV/BW.

Se ha evaluado el efecto del ketoprofeno sobre los valores hematológicos de 10 bovinos Jersey, machos, de 1 a 2 meses de edad, nacidos a nivel del mar (NM) y expuestos durante 30 días a 3 320m. de altitud (A), divididos en los grupos control (T1) y tratamiento con ketoprofeno (T2). Los resultados obtenidos a NM y a los 3 y 30 días de exposición en A fueron: Hematocrito (%) a NM: 28.50±5.09 y 26.60±4.10 para T1 y T2, respectivamente; al día 3 en A: 31.17±4.02 y 28.80±2.95 para T1 y T2, respectivamente y al día 30 en A: 39.50±4.28 y 38.00±2.94 para T1 y T2, respectivamente. Hemoglobina (g/dl) a NM: 8.37±1.63 y 7.96±1.33 para T1 y T2, respectivamente; al día 3 en A: 10.97±1.38 y 9.99±1.43 para T1 y T2, respectivamente y al día 30 en A: 13.72±1.09 y 14.04±1.68 para T1 y T2, respectivamente. Eritrocitos (x106/µl) a NM: 8.47±1.61 y 7.04±2.96 para T1 y T2, respectivamente; al día 3 en A: 8.41±1.52 y 8.99±2.54 para T1 y T2, respectivamente y al día 30 en A: 10.97±0.93 y 9.33±0.58 para T1 y T2, respectivamente. Volumen corpuscular medio (mm3) a NM: 34.03±4.90 y 41.20±11.13 para T1 y T2, respectivamente; al día 3 en A: 37.58±5.39 y 33.42±6.31 para T1 y T2, respectivamente y el día 30 en A: 36.17±4.39 y 41.01±5.61 para T1 y T2, respectivamente. Hemoglobina corpuscular media (ρg) a NM: 9.99±1.57 y 12.28±3.17 para T1 y T2, respectivamente; al día 3 en A: 13.27±2.19 y 11.50±2.04 para T1 y T2, respectivamente y al día 30 en A: 12.55±1.12 y 15.16±2.66 para T1 y T2, respectivamente. Concentración media de hemoglobina corpuscular (g/dl) a NM: 29.44±2.51 y 29.92±2.00 para T1 y T2, respectivamente; al día 3 en A: 35.23±1.34 y 34.56±2.02 para T1 y T2, respectivamente y al día 30 en A: 34.82±1.33 y 36.86±2.08 para T1 y T2, respectivamente. Leucocitos (x103/µl) a NM: 7.78±3.34 y 7.59±0.51 para T1 y T2, respectivamente; al día 3 en A: 12.90±3.43 y 12.00±1.88 para T1 y T2, respectivamente y al día 30 en A: 11.51±2.90 y 9.56±0.64 para T1 y T2, respectivamente. Se concluye que el tratamiento con ketoprofeno no tuvo efecto significativo (P>0.05) sobre los valores hematológicos de terneros Jersey, a los 30 días de exposición a 3 320m. de altitud. / The effect of ketoprofen upon the hematological values in 10 Jersey male calves from 1 to 2 months age, born at sea level (SL) and exposed at 3 320m. of altitude (A) during 30 days; they were divided in control (T1) and ketoprofen treatment (T2). The obtained values at SL and at the 3 and 30 days of exposure to A, were as follows: Hematocrit (%) at SL: 28.50±5.09 and 26.60±4.10 for T1 and T2 respectively; 3 days at A: 31.17±4.02 and 28.80±2.95 for T1 and T2, respectively and 30 days at A: 39.50±4.28 y 38.00±2.94 for T1 and T2, respectively. Hemoglobin (g/dl) at SL: 8.37±1.63 and 7.96±1.33 for T1 and T2 respectively; 3 days at A: 10.97±1.38 and 9.99±1.43 for T1 and T2, respectively and 30 days at A: 13.72±1.09 and 14.04±1.68 for T1 and T2, respectively. Erythrocytes (x106/µl) at SL: 8.47±1.61 y 7.04±2.96 for T1 and T2, respectively; 3 days at A: 8.41±1.52 y 8.99±2.54 for T1 and T2, respectively and 30 days at A: 10.97±0.93 and 9.33±0.58 for T1 and T2, respectively. Mean corpuscular volume (mm3) at SL: 34.03±4.90 and 41.20±11.13 for T1 and T2, respectively; 3 days at A: 37.58±5.39 and 33.42±6.31 for T1 and T2, respectively and 30 days at A: 36.17±4.39 and 41.01±5.61 for T1 and T2, respectively. Mean corpuscular hemoglobin (ρg) at SL: 9.99±1.57 and 12.28±3.17 for T1 and T2, respectively; 3 days at A: 13.27±2.19 and 11.50±2.04 for T1 and T2, respectively and 30 days at A: 12.55±1.12 and 15.16±2.66 for T1 and T2, respectively. Mean corpuscular hemoglobin concentration (g/dl) at SL: 29.44±2.51 and 29.92±2.00 for T1 and T2, respectively; 3 days at A: 35.23±1.34 and 34.56±2.02 for T1 and T2, respectively and 30 days at A: 34.82±1.33 and 36.86±2.08 for T1 and T2, respectively. Leukocytes (x103/µl) at SL: 7.78±3.34 and 7.59±0.51 for T1 and T2, respectively; 3 days at A: 12.90±3.43 and 12.00±1.88 for T1 and T2, respectively and 30 days at A: 11.51±2.90 and 9.56±0.64 for T1 and T2, respectively. In conclussion the ketoprofen treatment have not significative effect (P>0.05) upon the hematological values of Jersey calves, until 30 days of exposure to 3320m. of altitude.

Thesis (M.S.)--University of North Carolina at Wilmington, 2003. / Includes bibliographical references (leaves : [44]-51).

published_or_final_version / Physiology / Doctoral / Doctor of Philosophy