News - Part 12

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Discussion

We demonstrated that hyperosmolar solutions stimulated both mucin and ly-sozyme secretion, suggesting that either there is co-regulation of secretion or that both mucous and serous cells respond in the same way to the hyperosmolar agents. This is supported by the observation that the rate of mucin secretion was approximately two times that of lysozyme secretion without stimulation and under all conditions of stimulation. Read more »

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Evaluation of Secretory Pathways

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Evaluation of Secretory PathwaysThere was 47% increase in lysozyme secretion with exposure to 3.69 g/dL of saline solution and a 54% increase with exposure to 10.69 g/dL of saline solution (each n = 5, p < 0.05) compared with exposure to KHS (Fig 2). The amounts of lysozyme secretion in tracheae exposed to KHS, 3.69 g/dL of saline solution, and 10.69 g/dL of saline solution were 3.19 ± 0.38 ^g/g, 4.58 ± 0.37 ^g/g, and 4.68 ± 0.27 ^g/g, respectively. Read more »

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Statistical Analysis

Morphologic Evaluation of the Tracheal Epithelium
Tracheal segments (n = 4 each) were taken before and after immersion in 3.69 g/dL of saline solution (1,192 mOsm), 10.69 g/dL of saline solution (3,612 mOsm), or in KHS (288 mOsm). Tissue segments were immersed in 10% formalin and processed for light (wide-field) microscopy. Paraffin-embedded tissue was cut to 4-^m thickness, and slides were stained with hematoxylin and eosin. These were examined by a trained investigator masked as to treatment group (J.S.K.). He randomly selected eight fields for evaluation of epithelial damage. Read more »

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Hyperosmolar Solution

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Hyperosmolar SolutionThe amount of lysozyme secreted was expressed as lysozyme per trachea tissue weight (micrograms per gram).
Mucin Secretion With Only the Tracheal Lumen Exposed to Hyperosmolar Solution
To evaluate whether hyperosmolar solution stimulates tracheal secretion via the luminal side in this model, whole trachea were used. Read more »

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Lysozyme Secretion by Spectrophotometry

In brief, a 96-well microtiter plate was coated with 60 |j,L of DBA (6 |j,g/mL in phosphate-buffered saline solution [PBS]) and incubated at room temperature for 2 h. After rinsing with high-salt PBS (PBS containing 0.5 mol/L NaCl and 0.1% Tween 20), the plate was exposed to sample buffer and incubated at room temperature for 1 h. This was then treated with 50 |j,L of DBA conjugated with horseradish peroxidase (0.25 |j,g/mL) in PBS containing 1% bovine serum albumin. Read more »

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Mucin Secretion by Enzyme-Linked Lectin Assay

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Mucin Secretion by Enzyme-Linked Lectin AssayWe randomized the administration of agents to different segments in each experiment. There is anatomic variation in the amount of ferret tracheal mucin secretion, with significantly greater secretion on the proximal (cephalad) trachea than near the carina. We eliminated anatomic variation in secretion as a potential confounding variable by ensuring that each study group had an equal number of segments from each portion of the trachea. Each tracheal segment was exposed to only one test agent. Read more »

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea: Mucin and Lysozyme Secretion

Adult ferrets (1.7 to 2.3 kg, random sex) obtained from Marshall Farms (North Rose, NY) were killed by intraperitoneal injection of pentobarbital sodium (150 mg/kg body weight), and the trachea from the larynx to the carina was immediately removed. Each trachea was divided into four or eight roughly equal segments from the upper trachea to the carina. The segments were weighed and then immersed in 10 mL of Krebs-Henseleit solution (KHS) [containing 6.9 g/L of NaCl, 0.35 g/L of KCl, 0.16 g/L of KH2PO4, 0.141 g/L of MgSO4, 0.373 g/L of CaCl2, 2 g/L of D-glucose: pH 7.4, 255 mOsm/L]. Read more »

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret Trachea

Hyperosmolar Solutions Stimulate Mucus Secretion in the Ferret TracheaSputum induction using hypertonic saline solution inhalation has been used to obtain sputum specimens for diagnosing airway infection and for collecting secretions for research. More than 20 years ago, it was shown that when patients with chronic bronchitis inhaled a 7% (7 g/dL, 2,396 mOsm/L) saline aerosol, whole-lung mucociliary clearance (MCC) increased by 100% in the first 90 min over the clearance rate after inhalation of isotonic (0.9%) saline solution. More recently, patients with cystic fibrosis CF) were randomly assigned to receive either 0.9% saline or 6% saline by ultrasonic nebulization. Read more »

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: Summary

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: SummaryMicrobiology results suggested SI may be a useful tool in the multicentered research setting. Aliquots of induced sputum appear to provide a representative sample as quantitative bacterial cultures only varied by approximately a half-log count over time. In standard induced-sputum sample processing, di-thiothreitol has to be added to the sputum sample to homogenize it. However, the addition of dithiothre-itol decreases microbiological counts after 2 h. Thus, for multicenter studies, when microbiological samples may be sent to a centralized laboratory, it appears that a sample from the first 4-min aliquot may vary from the last aliquot by a half-log count. Read more »

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: Asthmatic subjects

SI was relatively well tolerated by all subjects, However, two subjects had a > 15% fall in FEVj with this procedure even when premedicated with albuterol, However, the adverse effects associated with the sedation and the procedure of bronchoscopy with BAL on pulmonary function and oxygenation in patients with CF is not well reported but would appear greater than those of SI. Throat irritation was felt by three subjects. Read more »

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