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Continuous transtracheal oxygen delivery during cardiopulmonary resuscitation. An alternative method of ventilation in a canine model.

Adequate oxygenation of apneic subjects can be maintained by constant flow transtracheal oxygen (TTO), but this method alone is associated with hypercapnia. The "bellows" effect of external chest compressions (ECC) might prevent this problem if the airway were kept open by TTO. In dogs, we investigated the utility of TTO delivered at 15 L/min by a percutaneously placed intratracheal catheter, plus ECC (TTO/ECC) as an alternative method of ventilation during CPR. TTO was applied to anesthetized, paralyzed dogs in normal sinus rhythm (NSR) at various rates of ECC and during ventricular fibrillation (VF) at an ECC rate of 80/min. During NSR and VF, hypercapnia did not develop and arterial oxygen saturations were maintained above 90 percent. During NSR, the PaCO2 decreased and the pH increased as the ECC rate increased. For many of the animals, coronary perfusion pressure remained above 20 mm Hg during VF, suggesting that these animals could be resuscitated to NSR. In another phase, after 15 min of VF using TTO/ECC, seven of nine animals were defibrillated. We conclude that ventilatory and hemodynamic support adequate to permit successful resuscitation to NSR is provided by the combination of TTO/ECC to apneic dogs during VF.

Animals↗

Lung response to particulates with emphasis on asbestos and other fibrous dusts.

Many theories have been proposed to explain asbestosis and asbestos-related pulmonary disease. However, none of the theories give a completely plausible explanation for the pathogenesis. Recently, attention has been drawn to a theory that the fibrogenicity or carcinogenicity of fibrous dust particles is related to fiber diameter and length rather than to chemical properties. This theory may help partially elucidate the disease process but is still far from solving the enigma of pulmonary fibrosis or carcinogenesis. The theory cannot explain the absence of these pathological effects among fiberglass workers or experimental animals exposed by inhalation (even though mesotheliomas are induced by intrapleural implantation and fiber dimension-related fibrogenicity is demonstrated by intratracheal injection). Little information regarding the pulmonary response to manmade fibrous particles is available in animals following inhalation exposure. Attempts should be made to confirm the absence of adverse effects using animal inhalation experiments even though to this point there is no conclusive evidence that either lung cancer or pulmonary diseases can be produced among employees in manmade fiber industries. A new research trend seems concentrated on testing the durability of asbestos or manmade fibers. This is based on the concept that biological effects of fibrous particles are the result of relative durability and that particles which can be fragmented or shortened may be less pathogenic. In the last two decades, considerable understanding about pulmonary fibrosis and carcinogenesis of asbestos has been achieved by clinical and animal experiments. In vitro tests including cytotoxicity, hemolysis, immunology, and enzyme biochemistry have provided important information on the interrelationships among these various biological effects of asbestos.

Animals↗

Magnesium oxide as carrier dust in benzo(a)pyrene-induced lung carcino-genesis in Syrian hamsters.

The neoplastic progression induced by intratracheal instillation of benzo[a]pyrene (BP) and magnesium oxide (MgO) was compared with that induced by intratracheal instillation of BP and ferric oxide (Fe2O3). BP and MgO produced squamous cell carcinomas and papillomas in the larynx with a latent period as shor as 9 weeks. They also induced many papillomas as well as squamous cell carcinomas and adenocarcinomas in the trachea and a papilloma, squamous cell carcinomas, adenocarcinomas, adenosquamous lesions, and peripheral adenomatoid lesions in the bronchi. They rarely caused tumors in other organs; only a few forestomach papillomas, one melanoma on the dorsal skin, and one ovarian carconoma were seen BP, with Fe2O3 as the carrier, induced a comparable number of histologically similar tumors; however, tumors developed more frequently in the main bronchi. Thus MgO strongly facilitated the tumor-inducing effects of BP, causing tumors in different areas of the respiratory tract, and was as effective as Fe2O3 as a carrier agent in the experimental induction of respiratory tumors.

Animals↗

DNA adduct formation by the environmental contaminant 3-nitrobenzanthrone after intratracheal instillation in rats.

3-Nitrobenzanthrone (3-NBA) is an environmental pollutant and suspected human carcinogen found in emissions from diesel and gasoline engines and on the surface of ambient air particulate matter; human exposure to 3-NBA is likely to occur primarily via the respiratory tract. In our study female Sprague Dawley rats were treated by intratracheal instillation with a single dose of 0.2 or 2 mg/kg body weight of 3-NBA. Using the butanol enrichment version of the (32)P-postlabeling method, DNA adduct formation by 3-NBA 48 hr after intratracheal administration in different organs (lung, pancreas, kidney, urinary bladder, heart, small intestine and liver) and in blood was investigated. The same adduct pattern consisting of up to 5 DNA adduct spots was detected by thin layer chromatography in all tissues and blood and at both doses. Highest total adduct levels were found in lung and pancreas (350 +/- 139 and 620 +/- 370 adducts per 10(8) nucleotides for the high dose and 39 +/- 18 and 55 +/- 34 adducts per 10(8) nucleotides for the low dose, respectively) followed by kidney, urinary bladder, heart, small intestine and liver. Adduct levels were dose-dependent in all organs (approximately 10-fold difference between doses). It was demonstrated by high performance liquid chromatography (HPLC) that all 5 3-NBA-derived DNA adducts formed in rats after intratracheal instillation are identical to those formed by other routes of application and are, as previously shown, formed from reductive metabolites bound to purine bases. Although total adduct levels in the blood were much lower (41 +/- 27 and 9.5 +/- 1.9 adducts per 10(8) nucleotides for the high and low dose, respectively) than those found in the lung, they were related to dose and to the levels found in lung. These results show that uptake of 3-NBA by the lung induces high levels of specific DNA adducts in several organs of the rat and an identical adduct pattern in DNA from blood. Therefore, 3-NBA-DNA adducts present in the blood are useful biomarkers for exposure to 3-NBA and may help to assess the effective biological dose in humans exposed to it.

Animals↗

Verification of endotracheal tube placement by prehospital providers: is a portable fiberoptic bronchoscope of value?

INTRODUCTION: This study was designed to examine whether a handheld, battery-operated fiberoptic bronchoscope (FOB) used to verify endotracheal tube (ETT) placement would be as sensitive and specific as other modes and whether a combination of multiple modes would further enhance the sensitivity and specificity of ETT placement verification. SETTING: An academic hospital-based air medical program. METHODS: This was a prospective, randomized study examining surgical patients undergoing general endotracheal anesthesia. Eighteen critical care transport (CCT) nurses, previously unfamiliar with FOB, were asked to identify intratracheal and intraesophageal ETTs by using misting, end-tidal carbon dioxide concentration (ETCO(2)), and FOB alone or with a combination of all three modes. The sensitivity and specificity of single and multimode verification were calculated and compared. RESULTS: Comparison of ETT verification by single mode alone revealed a rank order of sensitivity with ETCO(2) (0.97) > FOB (0.87) > misting (0.84), whereas all three modes had similar specificities (0.93-0.94). However, the use of the three-mode combination revealed a sensitivity and specificity of 1.0. CONCLUSIONS: As a single mode for ETT verification, use of a handheld, battery-operated FOB device allowed for direct visualization and had an 87% sensitivity and 93% specificity. When combined with misting and ETCO(2), FOB allowed 100% successful verification of ETT placement.

Air Ambulances↗

DNA fragmentation, DNA-protein crosslinks, postlabeled nucleotidic modifications, and 8-hydroxy-2'-deoxyguanosine in the lung but not in the liver of rats receiving intratracheal instillations of chromium(VI). Chemoprevention by oral N-acetylcysteine.

An in vivo study was carried out with the objectives of evaluating (a) the localization of DNA lesions resulting from exposure to chromium(VI) by the respiratory route, (b) the molecular nature of DNA alterations, and (c) modulation of DNA damage by a known chemopreventive agent. To this purpose, Sprague-Dawley rats received intratracheal instillations of sodium dichromate (0.25 mg/kg body weight) for three consecutive days, and the day after the last treatment lung and liver were removed for DNA purification. The results showed a selective localization of DNA lesions in the lung but not in the liver, which can be ascribed to toxicokinetics and metabolic characteristics of chromium(VI). DNA alterations included DNA-protein crosslinks, DNA fragmentation, nucleotidic modifications, and 8-hydroxy-2'-deoxyguanosine. The last two endpoints were evaluated, for the first time in chromium toxicology, by means of postlabeling procedures. This methodology was adapted to the detection of the DNA damage produced by those reactive oxygen species which result from the intracellular reduction of chromium(VI). The oral administration of the thiol N-acetylcysteine completely prevented any induction of DNA lesions in lung cells.

8-Hydroxy-2'-Deoxyguanosine↗

Morphologic and biochemical effects of intratracheally administered oil shale in rats.

Young adult rats were given three weekly intratracheal instillations of 30 mg of raw oil shale or spent oil shale suspended in 1.0 ml sterile physiological saline. Positive control groups received similar instillations of 30 mg or 5 mg of quartz. Animals were sacrificed and tissue samples taken for histopathology and biochemical analyses at 3 weeks, 7 weeks, 4 months, and 8 months following the first instillation. Rats exposed to raw shale, spent shale or quartz had increased lung weights compared with controls. Microscopically, all exposed groups developed granulomatous pneumonia and alveolar lipoproteinosis; pulmonary fibrosis was most severe in the quartz-exposed groups and progressed with time in these groups. Total amounts of pulmonary hydroxyproline, prolyl hydroxylase, total protein, and lipid phosphate were increased in shale or quartz-exposed groups; however, concentration of these substances on a per gram of lung tissue basis was not different from control groups.

Animals↗

Ultrastructural analysis of pulmonary capillaries in the course of experimental lung emphysema.

Processes which occur within the capillaries of interalveolar septa play a major role in many lung diseases. In the present study, ultrastructural analysis of pulmonary capillaries was made at 7 days after intratracheal instillation of a proteolytic enzyme - papain, that is when macrophage cumulation within pulmonary tissue was maximal. Experimental animals (male Wistar rats) were injected with BCG vaccine, in doses that would stimulate the macrophage system to produce TNF-alpha. All the experimental groups, compared with the control, showed cumulation and increased adherence of monocytes, granulocytes and blood platelets to endothelium. Changes within endothelial cells manifesting their activation were observed. In the animals receiving papain or BCG, focal destruction of endothelial cells was found. The picture of vascular changes induced with papain and by simultaneous stimulation with BCG vaccine revealed a number of features similar to those found in the adult respiratory distress syndrome. The study proved usefulness of vascular perfusion as a method to analyze the vascular system, particularly in the processes progressing with activation and increased adhesion of inflammatory cells to the endothelium of pulmonary capillaries.

Animals↗

Exogenous lung surfactant: effect on radiographic appearance in premature infants.

At birth, premature infants of 25-29 weeks gestation, at high risk for development of neonatal respiratory distress syndrome (RDS), were given a single dose (90 mg) of calf lung surfactant extract (CLSE) by intratracheal instillation. The frequency and severity of RDS were assessed with use of a simple radiographic scoring system in which pulmonary parenchymal densities and the prominence of the air-bronchogram effect were used as indicators of widespread atelectasis. Radiographs were obtained in surfactant-treated and control infants within the first 90 minutes of life as part of an initial evaluation of their pulmonary status. Subsequent examinations were performed at less than 24 hours and less than 48 hours of age. Radiographic assessment of lung disease compared consistently with coordinated data on oxygen and mean airway pressure requirements of the infants. Both indicated a significantly decreased frequency and severity of RDS in the infants treated with surfactant. The results provide supporting evidence of the effectiveness of exogenous lung surfactant replacement in mitigating RDS in very premature infants.

Animals↗

Bleomycin-induced pulmonary injury in mice deficient in SPARC.

SPARC (secreted protein, acidic and rich in cysteine) is a component of the matrix that appears to regulate tissue remodeling. There is evidence that it accumulates in the lung in the setting of pulmonary injury and fibrosis, but direct evidence of its involvement is only now emerging. We therefore investigated the development of pulmonary fibrosis induced by bleomycin administered either intratracheally or intraperitoneally in mice deficient in SPARC. Bleomycin (0.15 U/mouse) given intratracheally induced significantly more pulmonary fibrosis in mice deficient in SPARC compared with that in wild-type control mice, with the mutant mice demonstrating greater neutrophil accumulation in the lung. However, in wild-type and SPARC-deficient mice given intraperitoneal bleomycin (0.8 U/injection x 5 injections over 14 days), the pattern and severity of pulmonary fibrosis, as well as the levels of leukocyte recruitment, were similar in both strains of mice. These findings suggest that the involvement of SPARC in pulmonary injury is likely to be complex, dependent on several factors including the type, duration, and intensity of the insult. Furthermore, increased neutrophil accumulation in the peritoneal cavity was also observed in SPARC-null mice after acute chemical peritonitis. Together, these data suggest a possible role for SPARC in the recruitment of neutrophils to sites of acute inflammation.

Animals↗

Anti-vascular endothelial growth factor gene therapy attenuates lung injury and fibrosis in mice.

Vascular endothelial growth factor (VEGF) is an angiogenesis factor with proinflammatory roles. Flt-1 is one of the specific receptors for VEGF, and soluble flt-1 (sflt-1) binds to VEGF and competitively inhibits it from binding to the receptors. We examined the role of VEGF in the pathophysiology of bleomycin-induced pneumopathy in mice, using a new therapeutic strategy that comprises transfection of the sflt-1 gene into skeletal muscles as a biofactory for anti-VEGF therapy. The serum levels of sflt-1 were significantly increased at 3-14 days after the gene transfer. Transfection of the sflt-1 gene at 3 days before or 7 days after the intratracheal instillation of bleomycin decreased the number of inflammatory cells, the protein concentration in the bronchoalveolar lavage fluid and with von Willebrand factor expression at 14 days. Transfection of the sflt-1 gene also attenuated pulmonary fibrosis and apoptosis at 14 days. Since the inflammatory cell infiltration begins at 3 days and is followed by interstitial fibrosis, it is likely that VEGF has important roles as a proinflammatory, a permeability-inducing, and an angiogenesis factor not only in the early inflammatory phase but also in the late fibrotic phase. Furthermore, this method may be beneficial for treating lung injury and fibrosis from the viewpoint of clinical application, since it does not require the use of a viral vector or neutralizing Ab.

Animals↗

Protection of cattle from bovine tuberculosis by vaccination with BCG by the respiratory or subcutaneous route, but not by vaccination with killed Mycobacterium vaccae.

Groups of cattle were vaccinated either with BCG Pasteur by the intratracheal or subcutaneous route or with killed Mycobacterium vaccae by the intradermal route and challenged intratracheally 54 days later with Mycobacterium bovis. Vaccination with BCG resulted in fewer animals developing tuberculous lesions and in a reduction in the number of lesions in the diseased animals compared with the unvaccinated group and the group vaccinated with M vaccae. None of the nine animals vaccinated intratracheally with BCG developed any tuberculous lung lesions after challenge with M bovis, but two of the nine animals from each of the groups dosed subcutaneously with low and medium doses of BCG developed lung lesions. There was little difference in protection against the M bovis challenge between the animals receiving the low dose (10(3) colony forming units, cfu) or medium dose (10(5) cfu) of subcutaneous BCG, but the medium dose of BCG produced stronger cell-mediated immune responses to bovine purified protein derivative (PPD) after vaccination. Vaccination intradermally with 10(9) heat-killed M vaccae did not protect cattle against an experimental challenge with M bovis and induced only weak cell-mediated immune responses to bovine PPD.

Animals↗

Role of CC chemokines (macrophage inflammatory protein-1 beta, monocyte chemoattractant protein-1, RANTES) in acute lung injury in rats.

The role of the CC chemokines, macrophage inflammatory protein-1 beta (MIP-1 beta), monocyte chemotactic peptide-1 (MCP-1), and RANTES, in acute lung inflammatory injury induced by intrapulmonary deposition of IgG immune complexes injury in rats was determined. Rat MIP-1 beta, MCP-1, and RANTES were cloned, the proteins were expressed, and neutralizing Abs were developed. mRNA and protein expression for MIP-1 beta and MCP-1 were up-regulated during the inflammatory response, while mRNA and protein expression for RANTES were constitutive and unchanged during the inflammatory response. Treatment of rats with anti-MIP-1 beta Ab significantly decreased vascular permeability by 37% (p = 0.012), reduced neutrophil recruitment into lung by 65% (p = 0.047), and suppressed levels of TNF-alpha in bronchoalveolar lavage fluids by 61% (p = 0.008). Treatment of rats with anti-rat MCP-1 or anti-rat RANTES had no effect on the development of lung injury. In animals pretreated intratracheally with blocking Abs to MCP-1, RANTES, or MIP-1 beta, significant reductions in the bronchoalveolar lavage content of these chemokines occurred, suggesting that these Abs had reached their targets. Conversely, exogenously MIP-1 beta, but not RANTES or MCP-1, caused enhancement of the lung vascular leak. These data indicate that MIP-1 beta, but not MCP-1 or RANTES, plays an important role in intrapulmonary recruitment of neutrophils and development of lung injury in the model employed. The findings suggest that in chemokine-dependent inflammatory responses in lung CC chemokines do not necessarily demonstrate redundant function.

Acute Disease↗

Selective blockade of NF-kappa B activity in airway immune cells inhibits the effector phase of experimental asthma.

Knockout mice studies have revealed that NF-kappaB plays a critical role in Th2 cell differentiation and is therefore required for induction of allergic airway inflammation. However, the questions of whether NF-kappaB also plays a role in the effector phase of airway allergy and whether inhibiting NF-kappaB could have therapeutic value in the treatment of established asthma remain unanswered. To address these issues, we have assessed in OVA-sensitized wild-type mice the effects of selectively antagonizing NF-kappaB activity in the lungs during OVA challenge. Intratracheal administration of NF-kappaB decoy oligodeoxynucleotides to OVA-sensitized mice led to efficient nuclear transfection of airway immune cells, but not constitutive lung cells and draining lymph node cells, associated with abrogation of NF-kappaB activity in the airways upon OVA provocation. NF-kappaB inhibition was associated with strong attenuation of allergic lung inflammation, airway hyperresponsiveness, and local production of mucus, IL-5, IL-13, and eotaxin. IL-4 and OVA-specific IgE and IgG1 production was not reduced. This study demonstrates for the first time that activation of NF-kappaB in local immune cells is critically involved in the effector phase of allergic airway disease and that specific NF-kappaB inhibition in the lungs has therapeutic potential in the control of pulmonary allergy.

Allergens↗

Comparison of the investigational drug, LY146032, with vancomycin in experimental pneumonia due to methicillin-resistant Staphylococcus aureus.

The efficacy of LY146032 was compared with that of vancomycin in experimental pneumonia due to methicillin-resistant Staphylococcus aureus (MRSA). Emphysematous hamsters were challenged intratracheally with MRSA and given LY146032 (20 mg/kg 24h), vancomycin (40 mg/kg/24h) or normal saline by subcutaneous injection. Following infection with 2 X 10(9) cfu, survival among antibiotic-treated animals was significantly greater than that of the control group (P less than 0.01 at 96 h); however, no significant difference in survival between the hamsters given LY146032 and vancomycin was seen. To evaluate the influence of the antibiotics on the rate of bacterial killing within the lungs (pulmonary clearance), animals were challenged with a high inoculum (1 X 10(9) cfu) or low inoculum (1 X 10(6) cfu). Animals treated with LY146032 demonstrated a significant advantage in pulmonary clearance versus controls at both inocula; however, animals treated with vancomycin showed a statistically significant increase in pulmonary clearance versus controls only at the lower inoculum. We conclude that in this experimental model, LY146032 was as effective as vancomycin in treating infection with MRSA.

Animals↗

Pulmonary clearance and toxicity of respirable gallium arsenide particulates intratracheally instilled into rats.

Gallium arsenide (GaAs) is an intermetallic compound that is recognized as a potential toxicological risk to workers occupationally exposed to its dust. Previous results have shown that rats intratracheally instilled with a fraction of GaAs particulates, characterized with a mean count diameter of 8.30 microns and a mean volume diameter of 12.67 microns, developed signs of systemic arsenic intoxication, pulmonary inflammation, and pneumocyte hyperplasia. The results of the present study confirm these findings and also show that a significantly smaller fraction of GaAs is a relatively more severe pneumotoxicant. Decreasing the particle mean count and mean volume diameter to 1.63 micron and 5.82 microns, respectively, increased the in vivo dissolution rate of GaAs, increased the severity of pulmonary lesions previously associated with GaAs exposure, and resulted in unique pathological sequelae in affected lung tissue. Pulmonary fibrosis, as indicated by analysis of lung 4-hydroxyproline content, was not considered statistically significant although histological examination of lung tissue revealed a mild fibrotic response. These results provide additional evidence that pulmonary exposure to respirable GaAs particulates is a potential health hazard in the semiconductor industry.

Animals↗

Morphologic detection and functional assessment of reconstituted normal alveolar macrophages in the lungs of SCID mice.

Alveolar macrophages (AMs) from immunocompetent animals were isolated from bronchoalveolar lavage and labeled with the fluorescent marker 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI). These AMs were administered intratracheally into mechanically ventilated SCID mice. From 1 to 28 days later, the recipient mice underwent bronchoalveolar lavage to isolate their AMs. To determine whether reconstituted AMs were still immunocompetent, the recovered AMs were assayed for their ability to phagocytose fluorescein-labeled zymosan-coated beads. After incubation with the beads, samples were assayed using a fluorescent-activated cell sorter to identify DiI-labeled reconstituted AMs, unlabeled resident AMs, and the proportion of these two groups undergoing phagocytosis. DiI-labeled AMs accounted for approximately 50% of all returned AMs. Additionally, the reconstituted AMs from normal BALB/c mice retained phagocytic activity compared with AMs from immunodeficient SCID mice. Reconstituted AMs demonstrated enhanced phagocytic activity compared with resident SCID AMs for up to 28 days following reconstitution. These results indicate that immunocompetent AMs can be successfully reconstituted into an immunodeficient host to partially restore alveolar host defense.

Animals↗

Intratracheal catheters as drug delivery systems.

Medication delivery into the lungs can be used to provide a high therapeutic index for agents targeted to specific lung diseases. In addition, the lung can be used as a portal of entry for other agents targeted to systemic diseases. Delivery of medications into the lung can be accomplished by either instillation or aerosolization. Instillation approaches are limited by the fluid volume that can be given safely, and instilled liquids distribute according to gravity. In contrast, aerosolization approaches can deliver larger volumes over longer periods and aerosols distribute according to ventilation. In the mechanically ventilated patient, externally generated aerosols have very poor lung delivery because the endotracheal tube functions as a barrier to aerosol passage. Novel aerosol generating systems at the ends of small-diameter catheters that can be placed into the trachea (or beyond) are being developed to address this. In vitro testing has shown these systems to be capable of producing appropriately sized particles, with high rates of lung deposition. These catheters could be coupled with tracheal gas insufflation systems, not only to deliver therapeutic aerosols but also to create water aerosols to supply necessary humidification during tracheal gas insufflation.

Administration, Inhalation↗