Nicole Ashpole

Increased oesophageal acid exposure at the beginning of the recumbent period is primarily a recumbent‐awake phenomenon

Background  A significant increase in oesophageal acid exposure during early recumbent period has been demonstrated.

Reassessment of the Principal Characteristics of Gastroesophageal Reflux During the Recumbent Period Using Integrated Actigraphy-Acquired Information

OBJECTIVES:Characterization of gastroesophageal reflux (GERD) events during the sleep period has been hampered by lack of any patient-friendly technique that allows accurate assessment of sleep duration and awakening time, without confining patients to a sleep laboratory. Our aim was to compare principal reflux characteristics during the upright, recumbent-awake, and recumbent-asleep periods as well as to determine the effect of sleep awakenings on the principal reflux characteristics of the recumbent-asleep period using novel technology that allows integration of recorded actigraphy data into collected pH information.METHODS:Patients with heartburn at least three times a week for the previous 3 months were invited to participate in this study. All participants were evaluated by the demographics and the GERD Symptom Checklist questionnaires. Thereafter, patients underwent ambulatory 24-h esophageal pH monitoring concomitantly with actigraphy. A novel technique was used to superimpose simultaneously recorded raw actigraphy data over pH data, resulting in more accurate information about reflux events during upright, recumbent-awake, recumbent-asleep, and conscious awakening periods as well as the relationship between symptoms and acid reflux events in the aforementioned periods.RESULTS:Thirty-nine subjects (M/F: 26/13, mean age 56.6±14 years) with an abnormal pH test were enrolled into the study. The recumbent period appeared heterogeneous and was clearly divided into recumbent-awake (123.0±20.2 min) and recumbent-asleep (485.6±23.6 min) periods. The percent total time pH<4, the mean number of acid reflux events, and the number of symptoms associated with reflux events were significantly greater in the recumbent-awake as compared with the recumbent-asleep period. The mean duration of an acid reflux event was not different among upright, recumbent-awake, and recumbent-asleep periods. However, short-duration reflux events during the sleep period were associated with conscious awakenings as compared with those during sleep (0.74±0.11 min vs. 1.64±0.3 min, P=0.01).CONCLUSIONS:The recumbent period is divided into recumbent-awake and recumbent-asleep periods. The recumbent-awake period has significantly different principal reflux characteristics than the recumbent-asleep period. Duration of an acid reflux event during the recumbent-asleep period is not uniformly prolonged. Short-duration acid reflux events during the sleep period are likely due to conscious awakenings.

Visualization of conventional outflow tissue responses to netarsudil in living mouse eyes

Visual impairment due to glaucoma currently impacts 70 million people worldwide. While disease progression can be slowed or stopped with effective lowering of intraocular pressure, current medical treatments are often inadequate. Fortunately, three new classes of therapeutics that target the diseased conventional outflow tissue responsible for ocular hypertension are in the final stages of human testing. The rho kinase inhibitors have proven particularly efficacious and additive to current therapies. Unfortunately, non-contact technology that monitors the health of outflow tissue and its response to conventional outflow therapy is not available clinically. Using optical coherence tomographic (OCT) imaging and novel segmentation software, we present the first demonstration of drug effects on conventional outflow tissues in living eyes. Topical netarsudil (formerly AR-13324), a rho kinase/ norepinephrine transporter inhibitor, affected both proximal (trabecular meshwork and Schlemms Canal) and distal portions (intrascleral vessels) of the mouse conventional outflow tract. Hence, increased perfusion of outflow tissues was reliably resolved by OCT as widening of the trabecular meshwork and significant increases in cross-sectional area of Schlemms canal following netarsudil treatment. These changes occurred in conjunction with increased outflow facility, increased speckle variance intensity of outflow vessels, increased tracer deposition in conventional outflow tissues and decreased intraocular pressure. This is the first report using live imaging to show real-time drug effects on conventional outflow tissues and specifically the mechanism of action of netarsudil in mouse eyes. Advancements here pave the way for development of a clinic-friendly OCT platform for monitoring glaucoma therapy.

Caveolin-1 modulates intraocular pressure: implications for caveolae mechanoprotection in glaucoma

Polymorphisms in the CAV1/2 genes that encode signature proteins of caveolae are associated with glaucoma, the second leading cause of blindness worldwide, and with its major risk factor, intraocular pressure (IOP). We hypothesized that caveolin-1 (Cav-1) participates in IOP maintenance via modulation of aqueous humor drainage from the eye. We localize caveolae proteins to human and murine conventional drainage tissues and show that caveolae respond to mechanical stimulation. We show that Cav-1-deficient (Cav-1−/−) mice display ocular hypertension explained by reduced pressure-dependent drainage of aqueous humor. Cav-1 deficiency results in loss of caveolae in the Schlemm’s canal (SC) and trabecular meshwork. However, their absence did not appear to impact development nor adult form of the conventional outflow tissues according to rigorous quantitative ultrastructural analyses, but did affect cell and tissue behavior. Thus, when IOP is experimentally elevated, cells of the Cav-1−/− outflow tissues are more susceptible to plasma membrane rupture indicating that caveolae play a role in mechanoprotection. Additionally, aqueous drainage from Cav-1−/− eyes was more sensitive to nitric oxide (NO) synthase inhibition than controls, suggesting that excess NO partially compensates for outflow pathway dysfunction. These results provide a functional link between a glaucoma risk gene and glaucoma-relevant pathophysiology.

Physical Factors Affecting Outflow Facility Measurements in Mice.

PURPOSE Mice are commonly used to study conventional outflow physiology. This study examined how physical factors (hydration, temperature, and anterior chamber [AC] deepening) influence ocular perfusion measurements in mice. METHODS Outflow facility (C) and pressure-independent outflow (Fu) were assessed by multilevel constant pressure perfusion of enucleated eyes from C57BL/6 mice. To examine the effect of hydration, seven eyes were perfused at room temperature, either immersed to the limbus in saline and covered with wet tissue paper or exposed to room air. Temperature effects were examined in 12 eyes immersed in saline at 20 °C or 35 °C. Anterior chamber deepening was examined in 10 eyes with the cannula tip placed in the anterior versus posterior chamber (PC). Posterior bowing of the iris (AC deepening) was visualized by three-dimensional histology in perfusion-fixed C57BL/6 eyes and by spectral-domain optical coherence tomography in living CD1 mice. RESULTS Exposure to room air did not significantly affect C, but led to a nonzero Fu that was significantly reduced upon immersion in saline. Increasing temperature from 20 °C to 35 °C increased C by 2.5-fold, more than could be explained by viscosity changes alone (1.4-fold). Perfusion via the AC, but not the PC, led to posterior iris bowing and increased outflow. CONCLUSIONS Insufficient hydration contributes to the appearance of pressure-independent outflow in enucleated mouse eyes. Despite the large lens, AC deepening may artifactually increase outflow in mice. Temperature-dependent metabolic processes appear to influence conventional outflow regulation. Physical factors should be carefully controlled in any outflow studies involving mice.

The Ability of Nitric Oxide to Lower Intraocular Pressure Is Dependent on Guanylyl Cyclase.

Purpose While nitric oxide (NO) donors are emerging as treatments for glaucoma, the mechanism by which NO lowers intraocular pressure (IOP) is unclear. NO activates the enzyme guanylyl cyclase (GC) to produce cyclic guanosine monophosphate. We studied the ocular effects of inhaled and topically applied NO gas in mice and lambs, respectively. Methods IOP and aqueous humor (AqH) outflow were measured in WT and GC-1α subunit null (GC-1−/−) mice. Mice breathed 40 parts per million (ppm) NO in O2 or control gas (N2/O2). We also studied the effect of ocular NO gas exposure (80, 250, 500, and 1000 ppm) on IOP in anesthetized lambs. NO metabolites were measured in AqH and plasma. Results In awake WT mice, breathing NO for 40 minutes lowered IOP from 14.4 ± 1.9 mm Hg to 10.9 ± 1.0 mm Hg (n = 11, P < 0.001). Comparable results were obtained in anesthetized WT mice (n = 10, P < 0.001). In awake or anesthetized GC-1−/− mice, IOP did not change under similar experimental conditions (P ≥ 0.08, n = 20). Breathing NO increased in vivo outflow facility in WT but not GC-1−/− mice (+13.7 ± 14.6% vs. −12.1 ± 9.4%, n = 4 each, P < 0.05). In lambs, ocular exposure to NO lowered IOP in a dose-dependent manner (−0.43 mm Hg/ppm NO; n = 5 with 40 total measurements; P = 0.04) without producing corneal pathology or altering pulmonary and systemic hemodynamics. After ocular NO exposure, NO metabolites were increased in AqH (n = 8, P < 0.001) but not in plasma. Conclusions Breathing NO reduced IOP and increased outflow facility in a GC-dependent manner in mice. Exposure of ovine eyes to NO lowers IOP.

The effect of antireflux treatment on patients with gastroesophageal reflux disease undergoing a mental arithmetic stressor.

Background  Acute stress exacerbates heartburn in gastroesophageal reflux disease (GERD) patients by enhancing the perceptual responses to intraesophageal acid. The aim of the study was to determine if antireflux treatment can still alter stimulus response functions to acid in patients undergoing acute stress as compared with placebo.

Oesophageal sensation in response to high PCO2 and acidic solutions in nonerosive reflux disease

Eur J Clin Invest 2011