News - Part 13

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: Inflammatory Mediators

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: Inflammatory MediatorsThere was a mean half-log decrease in both nonmucoid P aeruginosa and S aureus with time (p = 0.05 and p < 0.01, respectively). The mean changes in nonmucoid P aeruginosa density and S aureus were 0.0.54 log10 cfu/mL (95% CI, — 1.60 to 0.52) and — 0.55 log10 cfu/mL (95% CI, — 1.42 to 0.32), respectively. There was no change in bacterial density of mucoid P aeruginosa (p = 0.12). R pickettii was seen in 60% of subjects but at low colony counts up to 7,600 colonies per milliliter.
IL-8 was detected in all subjects and in all samples. The mean ± SD concentration (log10) was 1.8 ± 1.4 ng/mL, 1.8 ± 1.5 ng/mL, 1.9 ± 1.6 ng/mL, 1.8 ± 1.6 ng/mL, and 1.8 ± 1.5 ng/mL at the five time points, respectively. The mean change in IL-8 (log10) was — 0.026 ng/mL (95% CI, — 0.103 to 0.051). There was no linear time trend in IL-8 concentrations (p = 0.99; Fig 3). Read more »

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

Urea was detected in all subjects and in all samples. The mean urea concentration was 4 ± 1 mg/dL, 3 ± 1 mg/dL, 3 ± 1 mg/dL, 4 ± 1 mg/dL, and 3 ± 1 mg/dL at the five time points. The mean change from the 0- to 4-min sample to the 16- to 20-min sample was 0.20 mg/dL (95% CI, — 0.65 to 0.25). There was no linear trend in urea concentration with time (p = 0.99).
Total Cell Counts and Cell Differentials
The total cell counts of samples collected at 4, 8, 12, 16, and 20 min (mean ± SEM) were 1.8 ± 1.6 X 107/mL, 1.8 ± 1.8 X 107/mL, 1.8 ± 1.7 X 107/ mL, 1.6 ± 1.7 X 107/mL, and 1.8 ± 1.8 X 107/mL, respectively (Fig 1). The mean change was 0.016 log10 cells per milliliter (95% CI, — 0.193 to 0.225). The nonsquamous cell counts per sample were 1.8 ± 1.7 X 107/mL, 1.7 ± 1.8 X 107/mL, 1.8 ± 1.7 X 107/mL, 1.6 ± 1.7 X 107/mL, and 1.9 ± 1.8 X 107/ mL, respectively. Read more »

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: Sample Size Calculations

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: Sample Size CalculationsThe null hypothesis for these tests was that sampling times, taken together, had no effect on the outcome of interest. A test for linear time trend was also performed for each outcome measure by fitting a regression in which time was modeled as a continuous variable. Finally, the mean change calculated as the 20-min value minus the 4-min value with 95% confidence intervals (CIs) was calculated.
This was a pilot study. There was no prior information from which a sample size calculation could be made. However, we performed a post hoc power analysis to determine what size effect could have been missed given the sample and results of our pilot study. Read more »

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: Quantitative Culture

Samples were run in triplicate and compared to a standard curve. In order to ensure that dithiothreitol did not degrade IL-8, we added 100 ng and 500 ng IL-8 for 90 min to each sample from three patients (ie, total of 15 samples) and generated standard curves. Dithiothreitol did not degrade IL-8. Urea concentration was measured at the clinical laboratory of the University of Washington Medical Center. Surfactant protein (SP)-A and SP-D were measured with a standard enzyme-linked immunosorbent assay as reported previously. Human SP-A was measured with a kit using two monoclonal antibodies provided by the Teijin Institute of Bio-Medicine (Hino, Japan). Recombinant SP-D was used as the standard for SP-D. The enzyme-linked immunosorbent assay was based on a sandwich method, using two monoclonal antibodies against human SP-D, 6B2 and 7C6, which were prepared against human SP-D purified from BAL fluids of patients with pulmonary alveolar proteinosis. Read more »

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

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: SIUsing a modification of the protocol described by Fahy et al, modified by the Cystic Fibrosis Foundation Therapeutics Development Network, subjects inhaled nebulized sterile hypertonic saline (3%) solution for 20 min from an Ultra-Neb 99 ultrasonic nebulizer (DeVilbiss; Somerset, PA). This nebulizer generates particles of a mean mass median diameter of 4.5 μm and has an output of 2.4 mL/min. After expelling saliva, subjects were encouraged to cough, and sputum was collected at 4-min intervals. Read more »

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum: Materials and Methods

We have previously shown that SI may provide a more representative airway sample than BAL in patients with CF. When SI is compared with BAL and expectorated sputum (ES), SI had a lower percentage of squamous cells than ES, all three techniques had a similar nonsquamous cell differential count, quantitative microbiology correlated best between ES and SI, and cytokine measurements by all three techniques were similar once corrected for the dilution of BAL. Read more »

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced Sputum

Analysis of Sequential Aliquots of Hypertonic Saline Solution-Induced SputumHypertonic saline has recently been used as a form of airway clearance in patients with cystic fibrosis (CF),-2 and as a means to obtain a lower airway sample for clinical use from patients who are unable to expectorate spontaneously. However, in high concentration (10%), hypertonic saline solution has been shown to cause bronchospasm in some patients with CF. Although sputum induction (SI) has been recognized as a research tool for over a decade with other airway disease such as asthma and COPD, only very recently has SI has been introduced as an outcome measure in CF. Read more »

Contributions of Retinoids to the Generation and Repair of the Pulmonary Alveolus: Retinoids Influence Alveolar Formation

Contributions of Retinoids to the Generation and Repair of the Pulmonary Alveolus: Retinoids Influence Alveolar FormationWhen cultures of rat LIF are supplemented with ATRA, there is a 2.5-fold to threefold increase in the steady-state level of tropoelastin messenger RNA. Tropoelastin is the soluble precursor of elastin, a major component of the elastic fiber. In the lung, its production is normally limited to late gestational and early postnatal life (up to postnatal day 21 in the rat and 7 years in humans). The RA-mediated increase in tropoelastin messenger RNA results from an approximately twofold increase in elastin gene transcription, rather than an increase in tropoelastin messenger RNA stability. Retinoid-responsive elements within the first 2 kilobases upstream of the transcriptional start site are required for this effect, and multiple elements may be involved. review
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Effects of Nebulized Diethylenetetraamine-NONOate in a Mouse Model of Acute Pseudomonas aeruginosa Pneumonia: Conclusion

Effects of Nebulized Diethylenetetraamine-NONOate in a Mouse Model of Acute Pseudomonas aeruginosa Pneumonia: ConclusionThis suggests that nebulized nitrite exposure in mice with pneumonia-induced pulmonary inflammation could result in intrapulmonary NO-dependent oxidative (nitrosative) effects. However, as DETA-NO and exhausted DETA were equally antibacterial in vitro, and as NaNO2 had a minimal effect, it is likely that NO and nitrite derived from DETA-NO did not play an important role in the observed in vivo effects of DETA-NO in murine pneumonia. Canadian Family Pharmacy

The exposure of sham mice to high-dose (ie, 125 ^mol) DETA-NO was associated with a significant, albeit slight, increase in pulmonary inflammation. This proinflammatory effect may be due to NO itself or to the DETA nucleophile moiety. Indeed, the lack of attenuation of leukocyte infiltration in DETA-NO-exposed mice with pneumonia, despite the decreased pulmonary bacterial load, may be due to a coincident proinflammatory effect of NO or DETA. The proinflammatory effects of NO have been well-recognized. NO may contribute to tissue inflammation and cell injury through several mechanisms. One of the most important mechanisms is thought to be the generation of the potent oxidizing species, peroxynitrite, via interaction of NO with superoxide. Read more »

Effects of Nebulized Diethylenetetraamine-NONOate in a Mouse Model of Acute Pseudomonas aeruginosa Pneumonia: NONOates

When NONOates are administered IV, they have been shown to cause both systemic and pulmonary vasodilation in animals. The rate of NO release and the resulting degree of vasodilation are related to the chemistry of the nucleophile moiety of the individual NONOate compounds. The intratracheal instillation of NONOate has been shown to selectively dilate the pulmonary vasculature both in normal rats and in rats with U-46619-induced pulmonary hypertension. Aerosolized NONOate also has been shown to attenuate pulmonary hypertension and improve oxygenation in several animal models. Read more »

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