Acid-base regulation is vital for animals and while the inorganic carbon system largely determines body fluid pH, another potentially valuable acid-base pair is ammonia (NH4+/NH3). This study focuses on the American horseshoe crab (Limulus polyphemus), a phylogenetically ancient marine chelicerate with no published studies on its acid-base physiology. Physiological and molecular analyses indicate that Na+/K+-ATPase, Rhesus-protein (Rh), and carbonic anhydrase (CA) are involved in acid-base homeostasis and/or ammonia regulation. This likely occurs in the book gills, which consist of ultrastructurally distinct regions. The ventral half-lamella is ion-leaky and displayed high Rh-protein, cytoplasmic CA, and hyperpolarization-activated cyclic nucleotide-gated K+ channel mRNA expression levels, suggesting a specialization in facilitated CO2 and/or ammonia diffusion compared to the dorsal half-lamella. During hypercapnia acclimation, hemolymph acid-base status exhibited a compensated respiratory acidosis accompanied with signs of metabolic depression. Ammonia influx associated with high environmental ammonia acclimation was successfully counteracted, but induced modifications in acid-base homeostasis. / October 2016

The American alligator (Alligator mississippiensis) is an aquatic diving reptile with a periodic breathing pattern. Previous work has identified pulmonary stretch receptors (PSR), both rapidly- and slowly-adapting, and intrapulmonary chemoreceptors (IPCs) that modulate breathing patterns in alligators. The purpose of the present study was to identify the effects of prolonged lung inflation and deflation (simulated dives) on PSR and/or IPC firing characteristics in the alligator. The effects of airway pressure, hypercapnia, and hypoxia on dynamic and static responses of pulmonary stretch receptors (PSR) were studied in juvenile alligators (mean mass = 246 g) at 24°C. Receptor activity appeared to be a mixture of slowly-adapting PSRs (SARs) and rapidly-adapting PSRs (RARs) with varying thresholds and degrees of adaptation, but no CO2 sensitivity. Dives were simulated in order to character receptor activity before, during, and after prolonged periods of lung inflation and deflation. Some stretch receptors showed a change in dynamic response, exhibiting inhibition for several breaths after 1 min of lung inflation, but were unaffected by prolonged deflation. For SAR, the post-dive inhibition was inhibited by CO2 and hypoxia alone. These airway stretch receptors may be involved in recovery of breathing patterns and lung volume during pre- and post-diving behavior and apneic periods in diving reptiles. These results suggest that inhibition of PSR firing following prolonged inflation may promote post-dive ventilation in alligators.

Little is known of the temporal cerebral blood flow response to a chemical stimulus consisting of increased PCO2 measured over time. Currently, there is only one study suggesting multiple phases in the CBF-CO2 response. Time constants of middle cerebral artery blood velocity (MCAV) response to a change in PETCO2 have been reported to be between 3 and 99.4 s. We studied the MCAV response in 28 subjects (10 females) to a sustained +10 mmHg above baseline (10 min) acute increase of PETCO2. We found that there were three distinct MCAV response patterns among subjects. Additionally, the responses of males and females differed. These studies suggest that there are multiple overlapping mechanisms controlling the chemoresponse of cerebral blood vessels and that these mechanisms may differ between men and women.

Little is known of the temporal cerebral blood flow response to a chemical stimulus consisting of increased PCO2 measured over time. Currently, there is only one study suggesting multiple phases in the CBF-CO2 response. Time constants of middle cerebral artery blood velocity (MCAV) response to a change in PETCO2 have been reported to be between 3 and 99.4 s. We studied the MCAV response in 28 subjects (10 females) to a sustained +10 mmHg above baseline (10 min) acute increase of PETCO2. We found that there were three distinct MCAV response patterns among subjects. Additionally, the responses of males and females differed. These studies suggest that there are multiple overlapping mechanisms controlling the chemoresponse of cerebral blood vessels and that these mechanisms may differ between men and women.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1943. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 28-29).

Normal levels of CO₂ are responsible for the maintenance of approximately 30% of sympathetic neurogenic vascular tone in intact cats. The central medullary chemoreceptors have been implicated as the major source of this CO₂ dependent neurogenic vascular tone. However, it is possible that spinal cord CO₂ sensitivity could also have mediated a portion of neurogenic vascular tone. Cats with acute and chronic spinal transections can maintain near normal levels of systemic arterial blood pressure, and show cardiovascular and sympathetic reflex changes in response to a variety of stimuli. Thus, it seemed likely that the acute spinal cat could exhibit the spinal component of CO₂ mediated sympathetic neurogenic vascular tone. Therefore the effects of systemic CO₂ increases on the perfusion pressures of vascularly isolated hindlimbs autoperfused at constant flow (as an indication of vascular resistance and sympathetic vascular tone) were studied in the acute cervical spinal cat. The contributions of the lumbar sympathetic system and the adrenal glands to perfusion pressure responses to CO₂ were evaluated. Experiments were carried out in mongrel cats with acute cervical (C2) transections. Each cat had one leg denervated by cutting and stripping the lumbar sympathetic chain from L₁-L₇. In all cats each hindlimb was vascularly isolated and perfused with blood taken from the abdominal aorta. Bilateral adrenalectomy was performed on 8 animals. CO₂ administration for 5 minutes resulted in biphasic increases in the perfusion pressures of both legs which were designated peak1 (P1) and peak2 (P2). Increasing PCO₂ from 16 to 38mm Hg, and from 16 to 62mm Hg resulted in significant P1 and P2 responses of the innervated leg. This also resulted in a significant P2 response, and an observable but insignificant P1 response, of the denervated leg. Adrenalectomy reduced P1 and P2 responses of the innervated leg, and abolished the P1 response and reduced the P2 response of the denervated leg. In non-adrenalectomized cats increasing PCO₂ also resulted in a significant increase in systemic arterial pressure (SAP) with no changes in heart rate (HR). In adrenalectomized cats increasing PCO₂ resulted in an observable but non-significant increase in SAP and a significant decrease in HR. These results suggested that: 1) The P1 response was primarily a sympathetic neurogenic response to increased CO₂. 2) The P2 response was primarily a hormonal response to CO₂ in the denervated leg, and a combination of hormonal and sympathetic neurogenic responses to CO₂ in the innervated leg. 3) The adrenal glands were mostly involved in the P2 response to CO₂, but possibly had a small role in the P1 response. 4) Other non-adrenal vasoconstrictor hormones may have played a role in the P2 response to C0₂. 5) Likely, CO₂ initially activated the sympathetic system to directly increase neurogenic tone, perhaps by stimulating sympathetic afferent or efferent neurons, or hypothetical spinal chemosensitive regions. Progressively the adrenal and possibly other unidentified vasoconstrictor hormone systems became activated, either directly by CO₂ or indirectly by CO₂ mediated sympathetic activation. These hormone systems may have also played a role in CO₂ mediated maintenance of vascular tone. / Medicine, Faculty of / Cellular and Physiological Sciences, Department of / Graduate

Adaptation to environmental fluctuations, through sensing and appropriate physiological responses, is crucial to homeostasis. Neuroepithelial cells (NECs) are putative chemoreceptors resembling mammalian Type I (glomus) cells. They have been shown to respond in vitro to changes in O2, CO2, NH3 and pH. Cytosolic carbonic anhydrase (Ca17a) is thought to be involved in CO2 sensing owing to its presence in NECs. A mutant line of zebrafish (Danio rerio) lacking functional Ca17a was generated using CRISPR/Cas9 technology and used to assess the role of Ca17a in initiating the cardiorespiratory responses to elevated CO2 (hypercapnia). Unfortunately, the homozygous knockout mutants (ca17a-/-) did not survive longer than ~12-14 days post fertilization (dpf), restricting experiments to early developmental stages (4-8 dpf). Changes in ventilation (fV) and cardiac (fH) frequency in response to hypercapnia (1% CO2) in wild type (ca17a+/+), heterozygous (ca17a+/-) and ca17a-/- fish were used to investigate Ca17a-dependent CO2 sensing and downstream signalling. Wild type fish exhibited hyperventilation during hypercapnia as indicated by an increase in fV. In the ca17a-/- fish, the hyperventilatory response was attenuated markedly, but only at 8 dpf. Hypercapnic tachycardia was observed for all genotypes and did not appear to be influenced by the absence of Ca17a. Interestingly, ca17a-/- fish exhibited a significantly reduced resting fH¬. This effect of knockout became more pronounced as the fish aged. Anesthesia did not contribute to the decreased fH in the ca17a-/- fish, nor did changes in cardiac adrenergic or cholinergic tone, which were probed using propranolol (-adrenergic receptor blocker) or atropine (muscarinic receptor blocker). The decrease in resting fH was prevented (“rescued”) when ca17a-/- embryos were injected with ca17a mRNA. Collectively, the results of this thesis support a role for Ca17a in promoting hyperventilation during hypercapnia in larval zebrafish and suggest a previously unrecognized role for Ca17a in determining resting heart rate.

No description available.

Sepsis has been defined as a systemic response to an infection. With an incidence of 3 per 1000 population per year or about 750 000 cases a year, this syndrome ranks as the 10th leading cause of death in the United States (1). Increasing severity of sepsis correlates with increasing mortality, which rises from 30-40% for severe sepsis up to 40-60% for septic shock. This thesis examines the effectiveness of adjunctive therapies, including activated protein C, hypercapnia and acidosis, and sodium selenite, in a clinically relevant ovine model of septic shock. The results from these studies can provide valuable information for future clinical trials on sepsis.<p>This thesis is divided into four sections: 1) sepsis overview; 2) an autologous fecal peritonitis model in sheep and its evaluation; 3) the series of studies on adjunctive therapeutics; and 4) ongoing studies and future perspective.<p>In the first section, a broad overview gives a rough introduction to delineate many aspects of sepsis syndrome such as terminology, etiology, epidemiology, pathophysiology and current guidelines for management. Hemodynamics in sepsis are especially elaborated since these are major observations throughout the studies presented later.<p>In the second section, the general characteristics of the sepsis models used in this thesis are elucidated. Data on hemodynamics, lung mechanics, gas exchange, etc. are presented to feature the ovine peritonitis model. The results of laboratory examinations for hematology, coagulation, bacteriology, biochemistry and hormonology are also presented. And then, I review currently used sepsis models with regards to their advantages and disadvantages.<p>The third section discusses three studies with their objectives, the methods used, the major findings, and the potential clinical implications.<p>9<p>1) Beneficial effects of recombinant human activated protein C in experimental septic shock. Activated protein C has a multitude of beneficial effects in severe sepsis and septic shock, including anti-inflammation, anti-coagulation, profibrinolysis, anti-apoptosis and endothelial protection. A clinical Phase III trial demonstrated that the administration of recombinant human activated protein C improved survival in patients with severe sepsis. However, doubts on the protective effects of activated protein C have persisted and been refueled by the recently published negative trials in less severely ill patients and in children. In the light of these ambiguities and uncertainties, we reinvestigated the effects of activated protein C in experimental septic shock.<p>2) Acute hypercapnia improves indices of tissue oxygenation more than dobutamine in septic shock. Hepercapnia has been found to possess beneficial effects in diverse acute inflammatory states independent of protective lung mechanics. To prove the hypothesis that acute hypercapnia has similar or superior hemodynamic effects to those of a dobutamine infusion, which may be particularly relevant in the presence of hemodynamic instability associated with respiratory failure, we investigated the effects of hypercapnia, which induced by inspiring extrinsic carbon dioxide in experimental septic shock.<p>3) High bolus dose of sodium selenite prolongs survival in an ovine model of septic shock. Selenite has both pro- and anti-oxidant effects. The administration of high dose sodium selenite may improve survival in septic shock patients. The benefit may be greater with the administration of a bolus (to achieve higher concentrations) rather than a continuous infusion. To test this hypothesis, we examined the effects of a high dose bolus administration of sodium selenite in experimental septic shock.<p>The fourth and final section talks about currently ongoing studies and offers some perspective on future direction. / Doctorat en Sciences médicales / info:eu-repo/semantics/nonPublished

BACKGROUND:We tested the hypothesis that ventilatory drive in hypoxia and hypercapnia is inversely correlated with the number of hypopneas and obstructive apneas per hour of sleep (obstructive apnea hypopnea index, OAHI) in children.METHODS:Fifty children, 6 to 12 years of age were studied. Participants had an in-home unattended polysomnogram to compute the OAHI. We subsequently estimated ventilatory drive in normoxia, at two levels of isocapnic hypoxia, and at three levels of hyperoxic hypercapnia in each subject. Experiments were done during wakefulness, and the mouth occlusion pressure measured 0.1 seconds after inspiratory onset (P0.1) was measured in all conditions. The slope of the relation between P0.1 and the partial pressure of end-tidal O2 or CO2 (PETO2 and PETCO2) served as the index of hypoxic or hypercapnic ventilatory drive.RESULTS:Hypoxic ventilatory drive correlated inversely with OAHI (r = -0.31, P = 0.041), but the hypercapnic ventilatory drive did not (r = -0.19, P = 0.27). We also found that the resting PETCO2 was significantly and positively correlated with the OAHI, suggesting that high OAHI values were associated with resting CO2 retention.CONCLUSIONS:In awake children the OAHI correlates inversely with the hypoxic ventilatory drive and positively with the resting PETCO2. Whether or not diminished hypoxic drive or resting CO2 retention while awake can explain the severity of sleep-disordered breathing in this population is uncertain, but a reduced hypoxic ventilatory drive and resting CO2 retention are associated with sleep-disordered breathing in 6-12 year old children.