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The effects of IL-5 on airway physiology and inflammation in rats.

BACKGROUND: There is evidence that the cytokine IL-5 is a prominent feature of airway inflammation in asthma. OBJECTIVE: The aim of this study was to determine whether exogenous IL-5 could cause changes in lung physiology, the early and late airway response after antigen challenge, and airway inflammation in rats that do not have a propensity to develop these changes after sensitization and challenge. METHOD AND RESULTS: Intratracheal administration of IL-5 to ovalbumin sensitized Brown Norway SSN rats increased the airway responsiveness to methacholine (AHR) 20 hours after administration of IL-5 at the same time as an increase in neutrophils occurred in the lung lavage. This effect was dose dependent and was not caused by endotoxin. Concurrent intratracheal administration of 50 ng of anti-IL-5 monoclonal antibody with 10 microg of recombinant human IL-5 decreased the AHR and neutrophil influx. Pretreatment with 3 microg of IL-5 had no effect on the early and late airway response or on AHR after ovalbumin challenge. However, IL-5 increased lung re-sistance 20 hours after antigen challenge. Although total lung cells and differential counts did not differ significantly 8 hours after antigen challenge, the blood lymphocyte CD4/CD8 ratio decreased in IL-5 pretreated rats (P <.05). In addition, in situ hybridization showed a significant increase in cells within the airway wall expressing IL-4 and IL-5 mRNA in IL-5 treated/challenged rats compared to controls (P <.05). CONCLUSION: The intratracheal administration of IL-5 causes only part of the physiologic changes that are associated with asthma. Other factors are necessary to obtain the complete asthma phenotype.

Animals↗

Interleukin-1-induced lung neutrophil accumulation and oxygen metabolite-mediated lung leak in rats.

We found that intratracheal administration of recombinant interleukin-1 alpha (IL-1) into rats rapidly (< 5 h) increased neutrophils in lung lavages and caused an acute edematous lung injury which was reflected by lung albumin accumulation (lung leak) and histological abnormalities (perivascular cuffing). These IL-1-dependent processes were inhibited by prior administration of recombinant IL-1 receptor antagonist and did not occur following administration of heated IL-1. Several lines of evidence suggested that neutrophil-derived oxygen metabolites contributed to lung leak. First, lung leak did not occur in rats rendered neutropenic by vinblastine treatment 4 days before IL-1 administration but did occur in neutrophil-replete rats given vinblastine 1 day before IL-1 administration and control rats given IL-1. Second, treatment with a hydroxyl radical scavenger, dimethyl sulfoxide (DMSO) or a superoxide anion scavenger, manganese superoxide dismutase, decreased lung leak, lung lavage neutrophils, and histological abnormalities in rats given IL-1 intratracheally. Third, intratracheal IL-1 administration increased lung oxidized glutathione (GSSG) levels and expired H2O2 concentrations, and these two indices of oxidative stress were decreased by dimethyl sulfoxide or manganese superoxide dismutase treatment. We conclude that intratracheal administration of IL-1 increases neutrophils in the lung and causes a neutrophil and oxygen metabolite-dependent acute edematous lung injury.

Animals↗

Intratracheal recombinant surfactant protein d prevents endotoxin shock in the newborn preterm lamb.

RATIONALE: The susceptibility of neonates to pulmonary and systemic infection has been associated with the immaturity of both lung structure and the immune system. Surfactant protein (SP) D is a member of the collectin family of innate immune molecules that plays an important role in innate host defense of the lung. OBJECTIVES: We tested whether treatment with recombinant human SP-D influenced the response of the lung and systemic circulation to intratracheally administered Escherichia coli lipopolysaccharides. METHODS: After intratracheal lipopolysaccharide instillation, preterm newborn lambs were treated with surfactant and ventilated for 5 h. MEASUREMENT: Survival rate, physiologic lung function, lung and systemic inflammation, and endotoxin level in plasma were evaluated. MAIN RESULTS: In control lambs, intratracheal lipopolysaccharides caused septic shock and death associated with increased endotoxin in plasma. In contrast, all lambs treated with recombinant human SP-D were physiologically stable and survived. Leakage of lipopolysaccharides from the lungs to the systemic circulation was prevented by intratracheal recombinant human SP-D. Recombinant human SP-D prevented systemic inflammation and decreased the expression of IL-1beta, IL-8, and IL-6 in the spleen and liver. Likewise, recombinant human SP-D decreased IL-1beta and IL-6 in the lung and IL-8 in the plasma. Recombinant human SP-D did not alter pulmonary mechanics following endotoxin exposure. Recombinant human SP-D was readily detected in the lung 5 h after intratracheal instillation. CONCLUSIONS: Intratracheal recombinant human SP-D prevented shock caused by endotoxin released from the lung during ventilation in the premature newborn.

Animals↗

Comparison of lower concentrations of lidocaine to suppress bucking before extubation during recovery of general anesthesia.

Thirty females, aged 25-50 years old, scheduled for abdominal total hysterectomy were randomly divided to two groups. For patients in group I, 2 mL 1% lidocaine solution was injected through the catheter on the tube to desensitize the trachea while surgeon started to close the peritoneum. In group II, 2 mL 2% lidocaine was used. The peak cuff pressures generated in the awakening group I patients were 49.0 +/- 2.5 cmH2 O (mean +/- SD) which shows greater than that in group II (27.9 +/- 7.2 cmH2 O), p less than 0.01. Bucking before awakening was also evaluated clinically. One hundred % (15/15) in group I patients bucked compared with 13.3% (2/15) in group II patients (p less than 0.01). Gag reflex was preserved in all patients and none suffered from aspiration postoperatively. Intratracheal administration of two mL 2% lidocaine through the catheter of modified endotracheal tube showed significant effect for suppressing bucking during recovery of general anesthesia.

Adult↗

Complications in the use of the subcutaneous tunnelled intratracheal oxygen catheter.

Transtracheal oxygen delivery seems to be a safe procedure in the treatment of chronic obstructive pulmonary disease (COPD) with chronic hypoxaemia. Even so, serious complications do occur. Three patients in whom we used a subcutaneous tunnelled intratracheal oxygen catheter (ITO2C) are described. Surgical intervention was required in all because of complications from the procedure. One of the complications--tracheal and catheter obstruction with stridor and subcutaneous emphysema by granulomatous tissue--has to our knowledge not been reported before.

Aged↗

Pulmonary delivery of free and liposomal insulin.

The effects of oligomerization and liposomal entrapment on pulmonary insulin absorption were investigated in rats using an intratracheal instillation method. The results indicated that both dimeric and hexameric insulins can be rapidly absorbed into the systemic circulation, producing a significant hypoglycemic response. Intratracheal instillation of insulin in two different oligomerized states has not resulted in any significant difference in the duration of hypoglycemic effect. However, the initial hypoglycemic response (first 10 min) obtained from intratracheal administration of 25 IU/kg hexameric insulin appears to be slower than that from the 25 IU/kg dimeric insulin, thereby suggesting that hexameric insulin may have a lower permeability coefficient across alveolar epithelium than the dimeric insulin. Intratracheal administration of insulin liposomes (dipalmitoylphosphatidyl choline:cholesterol, 7:2) led to facilitated pulmonary uptake of insulin and enhanced the hypoglycemic effect. Nevertheless, similar insulin uptake and pharmacodynamic response were obtained from both the physical mixture of insulin and blank liposomes and liposomally entrapped insulin.

Animals↗

Migration of dendritic cells into the draining lymph nodes of the lung after intratracheal instillation.

The migration of dendritic cell (DC)-enriched populations and alveolar macrophage (AM) populations isolated from PVG RT7.2 rats was studied after local administration to recipient PVG RT7.1 rats. The monoclonal antibody His41, which is directed against the common leukocyte antigen of the RT7.2 rat, was used to detect migrated cells. Injection of the splenic DC and AM subcutaneously into the footpads resulted in migration of both cell types to the popliteal lymph nodes after 24 h. DC located predominantly in the T cell-dependent areas, whereas AM located more in the medulla and medullary cords and spread throughout the outer cortex area. After intratracheal instillation of splenic DC, these cells were found predominantly in T cell-dependent areas of the draining lymph nodes of the lung after 24 h. In contrast, AM did not migrate to the draining lymph nodes after intratracheal instillation. Combined with those from earlier studies, these data show that DC present in the alveolar lumen may pick up airborne antigen and migrate to the draining lymph nodes of the lung, where they can induce primary T cell responses.

Animals↗

Pulmonary delivery of TH9507, a growth hormone releasing factor analogue, in the dog.

A modified growth hormone releasing factor (GRF; TH9507), a 44 amino acid peptide analogue of natural human growth hormone releasing factor, is being developed for the treatment of age-associated conditions resulting from diminished growth hormone (GH) secretion. The inhalation route of administration is being considered as an alternative to subcutaneous injection. A study was undertaken in dogs to investigate the absorption of TH9507 following pulmonary delivery. Male beagle dogs were administered TH9507 by intratracheal dry powder insufflation and subcutaneous injection at doses of approximately 375 and 38microg/kg, respectively. In a separate study, male and female dogs received 100microg/kg intravenously. Blood samples were collected at selected sampling times after dosing and plasma levels of TH9507 were measured by radioimmunoassay. The bioavailability by the inhaled route was 41% relative to subcutaneous dosing, with an absolute bioavailability estimated at 13%. No significant difference was observed for the terminal half-life of TH9507 after intratracheal (39min) and subcutaneous (26min) administrations. The mean residence time (MRT) was greater following intratracheal administration (74min versus 52min; P<0.01). These data indicate that the delivery of the TH9507 by the inhalation route may provide a suitable alternative to subcutaneous injection.

Absorption↗

Effect of intratracheal instillation of ultrafine carbon black on proinflammatory cytokine and chemokine release and mRNA expression in lung and lymph nodes of mice.

Our understanding of how ultrafine particles, which are constituents of particulate matter, affect immunological response is poor. To investigate the size-specific effect of ultrafine particles on pulmonary immune responses, translocation to lymph nodes, and chemokine mRNA expressions in lung and lymph nodes, we performed three experiments in 8-week-old male BALB/c mice. In experiment 1, we instilled 25 microg, 125 microg, or 625 microg of 14 nm carbon black (CB) particles intratracheally, once weekly for 4 weeks, and in experiment 2, we instilled 95 nm CB. For detection of total and differential cell counts and cytokine and chemokine protein release, we collected bronchoalveolar lavage (BAL) fluid 24 h after the last instillation of CB. Experiments 1 and 2 showed that 125 microg was the suitable dose for experiment 3, which we then performed on the same schedule and 4 h after the last instillation, we harvested the lung and mediastinal lymph node to detect chemokine mRNA expression by real-time RT-PCR. The total cell count as well as the differential cell counts such as macrophages, lymphocytes, and neutrophils in BAL fluid increased significantly in mice exposed to 14 nm CB in a dose-dependent manner. Release of cytokines such as interleukin (IL)-1beta, IL-6, and tumor necrosis factor-alpha increased significantly in BAL fluid in mice instilled with 14-nm CB. Macrophage inflammatory protein 1 alpha/CCL-3 protein and mRNA expression were increased significantly in the lungs and lymph nodes of mice given 14 nm CB. Histologically, deposition of CB was observed greater in the mediastinal lymph nodes of mice given 14 nm than in 95 nm CB. These findings indicate that repeated intratracheal instillation of ultrafine carbon black in mice leads to pulmonary inflammation, their translocation to mediastinal lymph nodes and increased chemokine mRNA expression in lung and lymph nodes size-specifically.

Animals↗

Lung antioxidant enzymes and cardiopulmonary responses in young rats exposed to hyperoxia and treated intratracheally with PEG catalase and superoxide dismutase.

The 27-day-old rat exposed to 100% oxygen (O2) for 8 days will have predictable lung vascular and parenchymal changes at 60 days of age. Using this model, the goals of this study are (1) to measure the lung antioxidant enzyme activities serially following intratracheal PEG antioxidant therapy during the 8-day O2 exposure; and (2) to assess chronic cardiopulmonary changes in the O2-exposed rats treated with PEG-CAT and/or PEG-CuZn SOD given intraperitoneally (IP) and/or intratracheally (IT). The study encompassed 202 male rats exposed to room air or oxygen. CuZn SOD doses were 300 U IT and 2000 U IP. The CAT dose was 500 or 4000 U IT and 10,000 U IP. At 60 days of age, the right ventricular systolic pressure (RVP), RV weight, % acinar wall arterial thickness, and parenchymal air space were significantly increased in O2-exposed rats compared to RA rats. The RVP, RV weight, and parenchymal changes were prevented by daily IT PEG-CAT 4000 U + CuZn SOD 300 U but the increased small artery muscularization was not. Three hours after the initial dose of IT PEG-CAT 4000 U, lung CAT activity was more than doubled and remained constant throughout the 8-day daily treatment course. This dose of CAT depressed the induction response to O2 of CuZn and MnSOD. It is concluded that daily intratracheal administration of PEG-CAT 4000 U + CuZn SOD 300 U can significantly ameliorate some of the chronic parenchymal and vascular lung O2 toxic changes. However, it appears that high-dose exogenous PEG-CAT suppresses the endogenous enzyme induction to hyperoxia of both CuZn and Mn-SOD.

Animals↗

[Exposure to ceramic fibers in the work environment. II. Occupational exposure to dust in plants producing ceramic fiber; fibrogenic effect of the fibers].

The aim of the work was to assess the exposure to dust containing ceramic fibres among workers employed in the plant producing aluminosilicate fibres, and to investigate fibrogenic properties of produced ceramic aluminosilicate fibres (L-1). The studies carried out in the work environment revealed that workers were exposed to dust containing respirable fibres from materials produced (fibres, mats). Mean concentrations of these fibres ranged from 0.07 to 0.37 f/cm3 and they were considerably lower than MAC value (1 f/cm3) proposed for respirable ceramic fibres. Mean total dust concentrations ranged from 0.4 to 2.9 mg/m3, and at two working posts they exceeded the proposed MAC value (2 mg/m3). In animal experiments (Wistar female rats) a single dose (25 mg) of L-1 fibres administered intratracheally induced pathological symptoms in lungs, limited to reactive changes in the form of granuloma with weakly pronounced cytolysis, and mean levels of hydroxyproline in the lungs did not differ significantly from the physiological level. The L-1 fibre was classified in the group of dusts with insignificant fibrogenic effects.

Animals↗

Comparative pulmonary toxicity of gallium arsenide, gallium(III) oxide, or arsenic(III) oxide intratracheally instilled into rats.

The relative toxicity of gallium arsenide (GaAs) and its metal oxides was assessed by intratracheally instilling particulate suspensions of GaAs (100 mg/kg), equimolar gallium as Ga2O3 (65 mg/kg), or a maximally tolerated nonlethal dose of arsenic as As2O3 (17 mg/kg). Two weeks after dosing, five rats from each group were randomly selected for the biochemical determination of lung lipid, protein, DNA, and collagen (4-hydroxyproline; 4-HP) content. The pulmonary retention of gallium and/or arsenic and the concentration of these metals in blood were also determined. Lungs from the remaining rats (n = 3) were examined histopathologically. Pulmonary exposure to Ga2O3 particulates significantly (p less than 0.05) increased the total lipid content of lungs relative to that observed in the vehicle-treated control animals. This response appeared to be associated with the pulmonary retention of gallium particulates (means = 36% of the gallium dose). In contrast, As2O3 particulates were not retained in the lung. Blood arsenic concentrations were 36 ppm which represented 20% of the total arsenic administered. Treatment with As2O3 significantly elevated lung dry weight as well as protein, DNA, and 4-HP content. These data suggest that As2O3 induced an acute fibrogenic response. The intratracheal instillation of GaAs particulates produced effects similar to those observed with the individual oxides. The total lung content of lipids, protein, and DNA was significantly elevated. These biochemical changes were accompanied by significant increases in lung dry weight and lung wet weight. Lungs from rats receiving GaAs particulates retained 44% of the dose as gallium and 28% of the dose as arsenic at the end of the 14-day study. Blood arsenic concentrations were 44 ppm (7% of the arsenic dose) while gallium was not detected in blood at this time. The primary histopathological observations 14 days after the intratracheal instillation of all metal particulates were an inflammatory response and pneumonocyte hyperplasia. The biological severity of these lesions, in descending order, was GaAs greater than As2O3 much greater than Ga2O3. It must be noted, however, that As2O3 was dosed at 0.25 X moles of GaAs.

Analysis of Variance↗

Light-guided tracheal puncture for percutaneous tracheostomy.

PURPOSE: To determine the effectiveness of lightwand-guided tracheal puncture for percutaneous tracheostomy. METHODS: The desired puncture site was marked on the skin of the anterior neck. A lightwand (Trachlight) was inserted into the patient's endotracheal tube (ETT), so that the number indicator on the lightwand matched the number indicator of the ETT of the patient. At this position, the light bulb of the lightwand was exactly placed at the tip of the endotracheal tube. With the lightwand turned on, the lightwand together with the endotracheal tube (ETT-LW) was slowly withdrawn from the trachea until a bright glow in the anterior neck could be seen 1 cm above the marked puncture site. At this position, the tip of the ETT was 1 cm above the puncture site. RESULTS: Percutaneous tracheostomy via a light-guided tracheal puncture was performed on 11 neurosurgical patients. The withdrawal of the endotracheal tube to a location above the puncture was accomplished easily with the lightwand. All percutaneous tracheostomies performed were successful, with ease and without any complications. The procedure time was 17.8 +/- 5.3 min. Mechanical ventilation was not interrupted during the whole procedure. CONCLUSION: The lightwand guided intratracheal puncture for percutaneous tracheostomy is a simple, effective, and safe procedure. This technique can avoid the risk of puncturing the endotracheal tube and/or cuff, thus allowing adequate ventilation and oxygenation during the percutaneous tracheostomy. Furthermore, this technique is inexpensive and minimizes the risk of damaging equipment like the fibreoptic bronchoscope.

Bronchoscopy↗

Emergency alternatives to intravenous access. Intraosseous, intratracheal, sublingual, and other-site drug administration.

Difficulties and delays in establishing intravenous access are not uncommon in emergency situations in pediatrics. Alternatives to venous cannulation exist, including intraosseous access, intratracheal drug administration, sublingual and intralingual injection, the intrapenile route, and intracardiac injection. Each of these emergency alternatives to intravenous access is discussed from the historical, technical, utilitarian, and risk-benefit aspects. It is concluded that the intraosseous effective alternative to intravenous access in emergency situations.

Administration, Sublingual↗

Prolonged partial liquid ventilation in spontaneously breathing awake animals.

OBJECTIVE: To date, studies of partial liquid ventilation (PLV) have examined its effects acutely in anesthetized and mechanically ventilated subjects. We set out to develop a model of prolonged PLV in awake, spontaneously breathing animals. DESIGN: Animal case series SETTING: Cardiopulmonary physiology laboratory. SUBJECTS: Fifteen New Zealand white rabbits (3.2+/-0.39 kg). INTERVENTIONS: Animals were anesthetized and instrumented with a novel technique allowing percutaneously assisted placement of an intratracheal catheter with a subcutaneously tunneled externalized free end. After anesthetic recovery, PLV was performed in spontaneously breathing unsedated animals. MEASUREMENTS AND MAIN RESULTS: Evaporative losses were determined using a single 10 mL/kg perflubron dose (n = 5); hourly radiographs were obtained until residual perflubron was minimal. For prolonged PLV (n = 10), a 10-mL/kg initial perflubron dose was followed every 4 hrs with 5-mL/kg supplements. Radiographs were obtained immediately before and after perflubron administration and were scored (0-5) by a radiologist blinded to dosing regimen and time interval. Data were analyzed with ANOVA and Student's t-test with correction for multiple comparisons. Initial filling was nearly complete (score = 4.8+/-0.42); lungs were maintained approximately half-filled through 4 hrs (score = 2.53+/-0.71). By 6 hrs, the majority of perflubron had evaporated (score = 1.75+/-0.53). Over 24 hrs, radiographs documented continuous perflubron exposure (postffill = 4.53+/-0.64, prefill = 3.40+/-0.71, average = 3.97+/-0.43); scores were comparatively higher after filling (after = 4.53+/-0.64, before = 3.4+/-0.71, p< .001). Left and right lung volumes were equivalent (left = 4.06+/-0.47, right = 3.89+/-0.39, p = .027). All animals survived the 24 hrs of PLV. CONCLUSIONS: Percutaneously assisted intratracheal cannulation with catheter exteriorization permits prolonged PLV in spontaneously breathing, unsedated animals. Continuous perfluorocarbon exposure with this method is reproducible, consistent, and well tolerated for 24 hrs in uninjured animals.

Analysis of Variance↗

In vivo mutagenesis induced by benzo[a]pyrene instilled into the lung of gpt delta transgenic mice.

Benzo[a]pyrene (B[a]P) is a ubiquitous airborne pollutant whose mutagenicity has been evaluated previously by oral and intraperitoneal administration to experimental animals. In this study, mutagenesis in the lungs, the target organ of air pollutants, was examined after a single intratracheal instillation of 0-2 mg B[a]P into gpt delta transgenic mice. Intratracheal injection of B[a]P resulted in a statistically significant and dose-dependent increase in gpt mutant frequency as measured by 6-thioguanine selection. The mutant frequencies at B[a]P doses of 0.5, 1, and 2 mg were 2.8, 4.2, and 6.8 times higher than the frequency seen in nontreated mice (0.60 +/- 0.13 x 10(-5)). The most frequent mutations induced by B[a]P treatment were G:C-->T:A transversions, which are characteristic of B[a]P mutagenesis in other models, and single-base deletions of G:C base pairs. To characterize the hotspots of B[a]P-induced mutations in the gpt gene, we analyzed sequences adjacent to the mutated G:C base pairs. Guanine bases centered in the nucleotide sequences CGT, CGA, and CGG were the most frequent targets of B[a]P. Our results indicate that intratracheal instillation of B[a]P into gpt delta mice causes a dose-dependent increase in gpt mutant frequency in the lung, and that the predominant mutation induced is G:C-->T:A transversion.

Air Pollutants↗

[Sterilization of intratracheal tubes].

After a brief review of classical and modern methods of sterilisation of endo-tracheal catheters, where they emphasize the dangers of the use of ethylene oxide, the authors recall the common procedure which they used: washing with water and soap, and soaking for 6 to 7 hours in formaldehyde or in a solution of quaternary ammonium salts, and all manipulations should be made using gloves.

1-Propanol↗

Intratracheal administration of endotoxin and cytokines. VII. The soluble interleukin-1 receptor and the soluble tumor necrosis factor receptor II (p80) inhibit acute inflammation.

Intratracheal administration of endotoxin (LPS) causes acute neutrophilic inflammation via induction of pulmonary tumor necrosis factor alpha (TNF) and interleukin-1 (IL-1) expression. In the present study, the anti-inflammatory activity of soluble IL-1 receptor (sIL-1r) and soluble TNF receptor p80 (sTNFr-p80) in LPS-induced acute pulmonary inflammation was investigated. The sIL-1r coinjected intratracheally with LPS in rats significantly inhibits neutrophilic exudation into bronchoalveolar lavage (BAL) fluid by 47% after 6 hr compared to injection of LPS alone. TNF and IL-6 in the same BAL fluids were both lowered by approximately 50% after intratracheal coinjection of sIL-1r and LPS as compared to LPS alone. In the same model, the sTNFr-p80 inhibited acute inflammation. Paradoxically, TNF levels in BAL fluids were generally elevated after the intratracheal coinjection of LPS and monomeric sTNFr-p80 compared to injection of LPS injection alone. The combined anti-inflammatory effect of sIL-1r and sTNFr-p80 at the maximally effective individual doses is not significantly greater than the effect of either soluble receptor alone.

Animals↗