Nam1p, a protein involved in RNA processing and translation, is coupled to transcription through an interaction with yeast mitochondrial RNA polymerase.

Alignment of three fungal mtRNA polymerases revealed conserved amino acid sequences in an amino-terminal region of the Saccharomyces cerevisiae enzyme implicated previously as harboring an important functional domain. Phenotypic analysis of deletion and point mutations, in conjunction with a yeast two-hybrid assay, revealed that Nam1p, a protein involved in RNA processing and translation in mitochondria, binds specifically to this domain. The significance of this interaction in vivo was demonstrated by the fact that the temperature-sensitive phenotype of a deletion mutation (rpo41Delta2), which impinges on this amino-terminal domain, is suppressed by overproducing Nam1p. In addition, mutations in the amino-terminal domain result specifically in decreased steady-state levels of mature mitochondrial CYTB and COXI transcripts, which is a primary defect observed in NAM1 null mutant yeast strains. Finally, one point mutation (R129D) did not abolish Nam1p binding, yet displayed an obvious COX1/CYTB transcript defect. This mutation exhibited the most severe mitochondrial phenotype, suggesting that mutations in the amino-terminal domain can perturb other critical interactions, in addition to Nam1p binding, that contribute to the observed phenotypes. These results implicate the amino-terminal domain of mtRNA polymerases in coupling additional factors and activities involved in mitochondrial gene expression directly to the transcription machinery.

History of high frequency oscillation.

The article describes the evolution of high frequency oscillation since its first use by Lunkenheimer through the initial failed NIH trial and subsequent more successful trials to its current widespread use in the neonatal population. The importance of oscillating at an optimal lung volume, achieved through a volume recruitment manoeuvre, is emphasised as is the efficacy with which oscillation clears CO2. The lack of adequate control of these two factors in the initial NIH trial is suggested as a possible cause of the trial's failure. Comment is made on optimising oscillator settings as well on elementary mechanics of high frequency oscillation and the effect of high frequency oscillation on surfactant degradation. Given the difficulty of recruiting lung volume in late RDS, a suggestion is made to combine high frequency oscillation with perfluorocarbon. The former as a mechanism for maintaining lung volume which has been recruited by the perfluorocarbon. The authors speculate that the use of high frequency oscillation will increase in both the paediatric and adult population.

The open lung during small tidal volume ventilation: concepts of recruitment and "optimal" positive end-expiratory pressure.

OBJECTIVES: To test the hypotheses that during small tidal volume ventilation (5 mL/kg) deliberate volume recruitment maneuvers allow expansion of atelectatic lung units and that a high positive end-expiratory pressure (PEEP) above the lower inflection point of the pressure/volume (PV) curve is not necessarily required to maintain recruited lung volume in acute lung injury. DESIGN: Prospective, randomized, controlled animal study. SETTING: An animal laboratory in a university setting. SUBJECTS: Adult New-Zealand rabbits. INTERVENTIONS: We studied a) the relationship of dynamic loops during intermittent positive pressure ventilation to the quasi-static PV curve, and b) the effect of lung recruitment on oxygenation, end-expiratory lung volume (EELV), and dynamic compliance in two groups (n = 4 per group) of lung-injured animals (lung lavage model): 1) the sustained inflation group, which received ventilation after a recruitment maneuver (sustained inflation); and 2) the control group, which received ventilation without any lung recruitment. MEASUREMENTS AND MAIN RESULTS: In the presence of PV hysteresis, a single sustained inflation to 30 cm H2O boosted the ventilatory cycle onto the deflation limb of the PV curve. This resulted in a significant increase in EELV, oxygenation, and dynamic compliance despite equal PEEP levels used before and after the recruitment maneuver. Furthermore, after a single sustained inflation, oxygenation remained high over 4 hrs of ventilation when a PEEP above the critical closing pressure of the lungs, defined as "optimal" PEEP, was used and was significantly higher compared with that in the control group ventilated at equal PEEP without preceding lung recruitment. CONCLUSIONS: The observation that ventilation occurs on the deflation limb of the tidal cycle-specific PV curve allows placement of the ventilatory cycle, by means of a recruitment maneuver, onto the deflation limb of the PV envelope of the optimally recruited lung. This strategy ensures sufficient lung volume recruitment to maintain the lungs during the tidal cycle while using relatively low airway pressures.

Concealed air leak associated with large tidal volumes in partial liquid ventilation.

Current ventilator strategies aim at maintaining an open lung and limiting both peak inspiratory pressures and tidal volumes to avoid alveolar distension. Perfluorocarbons, as well as being excellent solvents for oxygen and carbon dioxide, have the unique properties of being able to recruit dependent lung regions and improve pulmonary mechanics. Optimal ventilator strategies for partial liquid ventilation (PLV) have not yet been clearly defined. In the surfactant-depleted rabbit model, an approach involving a large tidal volume (VT) (15 ml/kg) and lung filled to FRC with perfluorocarbon (PFC) was compared with strategies involving a moderate VT (9 ml/kg) and partially filled lung (6 ml/kg), a moderate VT (9 ml/kg) and lung filled to FRC with PFC, and a large VT (15 ml/kg) and partially filled lung (6 ml/kg). PEEP was maintained at 5 cm H2O except in the moderate VT, partial-filling group, in which a PEEP of 9 cm H2O was used to maintain the rabbits for the duration of the experiment. Oxygenation was satisfactory in all groups, and peak inspiratory pressures were not significantly different. However, five of the 13 animals in the large-VT, PFC-filled lung group died of a pneumothorax prior to completion of the experiment. Of the eight animals in this group surviving the experiment, two had radiographic evidence of pneumothoraces, with an additional three animals having autopsy evidence of air leak. Of the 22 animals in the other groups, all survived with the exception of a single rabbit in the large VT, partial-filling group, which had both radiographic and autopsy evidence of air leak. We conclude that there is a significant risk of barotrauma in a PLV strategy in which a large VT is used in association with a lung filled to FRC with perfluorocarbon. Adequate gas exchange can be achieved with alternative ventilation strategies in combination with PLV.

The effect of single-dose and continuous skeletal muscle paralysis on respiratory system compliance in paediatric intensive care patients.

OBJECTIVE: To investigate the effect of single dose and continuous skeletal muscle paralysis on respiratory system compliance in 53 paediatric intensive care patients. DESIGN: Prospective clinical study. SETTING: Multidisciplinary paediatric intensive care unit. PATIENTS: Twenty-three children ventilated for acute pulmonary pathology, and 30 ventilated for isolated intracranial pathology, who initially had normal lungs. INTERVENTIONS: The 23 patients with acute pulmonary pathology received a single dose of muscle relaxant to facilitate diagnostic procedures. Fifteen patients with isolated intracranial pathology received continuous skeletal muscle paralysis for longer than 24 h, and the other 15 received no paralysis. MEASUREMENTS AND RESULTS: Respiratory system compliance deteriorated by 14% from 0.519 +/- 0.2 to 0.445 +/- 0.18 ml cmH2O-1 kg-1 (p < 0.001) following a single dose of muscle relaxant in the 23 patients with acute pulmonary pathology. In the 15 with isolated intracranial pathology who received continuous skeletal muscle paralysis there was a progressive deterioration in compliance, which reached 50% of the initial compliance by day 4 of paralysis (p < 0.001) and improved back to normal following discontinuation of paralysis. There were no changes in compliance in the 15 patients with isolated intracranial pathology who were ventilated but not paralysed. The paralysed patients required mechanical ventilation longer than the non-paralysed patients (p < 0.001), and 26% of these patients developed nosocomial pneumonia (p = 0.03), a complication that was not seen in the non-paralysed patients. CONCLUSIONS: Skeletal muscle paralysis results in immediate and progressive deterioration of respiratory system compliance and increased incidence of nosocomial pneumonia. The benefits of paralysis should be balanced against the risks of deteriorating pulmonary function.

Increased respiratory system resistance and bronchial smooth muscle hypertrophy in children with acute postoperative pulmonary hypertension.

Following surgery for congenital heart disease, there is often an increased reactivity of the pulmonary vasculature to stimuli, resulting in rapid increases in pulmonary artery pressure and a clinical impression of stiff lungs. Lung mechanics were measured in 30 children, mean age 6.7 +/- 4.1 mo, who were ventilated and had pulmonary artery pressure monitoring following surgery for congenital heart disease. A group of 15 patients developed postoperative pulmonary hypertension. In these patients, respiratory system resistance was 43% higher (p = 0.001) and compliance 11% lower (p = 0.004) during acute pulmonary hypertension compared with baseline pulmonary artery pressure. No changes in resistance or compliance were seen in the 15 patients who did not develop pulmonary hypertension. The changes in lung mechanics interfered with mechanical ventilation, resulting in a 9.4% rise in PaCO2 during pulmonary hypertension. The bronchial smooth muscle was found to be increased by 68%, and the vascular smooth muscle was more than twice normal in lung biopsies from 9 pulmonary hypertension patients compared with 6 age-matched postmortem controls patients who had no cardiac or pulmonary disease. The bombesin-immunoreactive pulmonary neuroendocrine cells (PNEC) were also increased in the pulmonary hypertension patients. These findings suggest a coconstriction and cohypertrophy of bronchial and vascular smooth muscle during pulmonary hypertension. Mediators, such as bombesin, endothelin-1, and serotonin, are known to be produced by PNEC and may be involved in the observed vasoconstriction, increased respiratory system resistance, and smooth muscle hypertrophy.

Maximal expiratory flows generated by rapid chest compression following end-inspiratory occlusion or expiratory clamping in young children.

Partial forced expiratory flow-volume curves obtained by the rapid chest compression technique are being widely used to assess pulmonary function in infants and young children. The aim of this study is to assess whether in this age group flow limitation is achieved with the partial forced expiratory flow-volume curve with rapid chest compression. In eight infants and young children sedated with chloral hydrate, flow-volume curves were obtained by regular rapid chest compression technique, end-inspiratory airway occlusion prior to rapid chest compression, and expiratory clamping prior to rapid chest compression. In each technique, beginning with a cuff pressure of 20 cmH2O, the cuff pressure was increased by 10 cmH2O increments until the compression pressure reached 90 cmH2O. Maximal flow-volume curves were generated by each technique. End-inspiratory occlusion prior to rapid chest compression caused higher flows over the entire phase of expiration than the regular rapid chest compression. This increase could be observed over the entire phase of expiration. Forced expiratory flow at 50% and at 75% of vital capacity (V50 and V75) with regular rapid chest compression were 207 +/- 44 ml.s-1 (mean +/- SD) and 138 +/- 59 ml.sec-1, respectively. When end-inspiratory occlusion preceded rapid chest compression, V50 and V75 increased to 283 +/- 114 and 206 +/- 61 ml.sec-1 respectively, with a mean increase in V50 of 34% and in V75 of 31%. When expiratory clamping preceded the compression, even higher expiratory flows resulted.(ABSTRACT TRUNCATED AT 250 WORDS)

Relationship between PaO2 and lung volume during high frequency oscillatory ventilation.

The relationship between oxygenation and lung volume during high frequency oscillatory ventilation (HFOV) was studied. We ventilated anesthetized, tracheostomized adult rabbits that were rendered surfactant-deficient by lung lavage. Lung volume was measured by the 'disconnection technique'. In the first experiment, HFOV was commenced after conventional mechanical ventilation (CMV) for 1 hr. In the absence of sustained inflation (SI), oxygenation improved with time during HFOV. In the second experiment, HFOV was instituted after CMV for 4 hr. In the absence of SI, all animals expired during the experimental period. In the third experiment we ventilated rabbits for 4 hr and then switched to HFOV. We applied SI first and increased mean airway pressure (MAP) by increments of 2 cmH2O every 15 min. However, there was little improvement in PaO2 despite the use of repeated SI and the increase in MAP. We conclude that oxygenation has a linear relationship to lung volume during HFOV, and that secondary lung injury due to long-term CMV impairs the response to HFOV. Therefore, it is important to minimize the risk of such secondary injuries before instituting HFOV.

Pulmonary hypertension after postlavage lung injury in rabbits: possible role of polymorphonuclear leukocytes.

Previous studies showed that repeated lung lavage leads to a severe lung injury with very poor gas exchange, a substantial protein leak into the alveoli with hyaline membrane formation, pulmonary hypertension, and migration of granulocytes (PMN) into the alveolar spaces. Depletion of PMN leads to a better gas exchange and a markedly decreased protein leak with only scanty hyaline membranes. In this study we show that there is sustained pulmonary hypertension after the lung lavage, but in PMN-depleted rabbits there is no postlavage increase in pulmonary arterial pressure. Changing the shunt fraction by manipulating mean airway pressure still leads to a hypoxic vasoconstriction with increase of pulmonary arterial pressure. Thus, after lung lavage, pulmonary reactivity to hypoxia is still preserved. Comparisons between high-frequency ventilation and conventional mechanical ventilation at the same mean airway pressures showed that equal mean airway pressure in these two very different modes of ventilation do not translate into the same mean functional lung volumes.

Lung injury in a surfactant-deficient lung is modified by indomethacin.

Repetitive total lung lavage in adult rabbits leads to a reproducible severe surfactant-deficient lung injury. Hypoxemia requiring mechanical ventilation occurs, accompanied by a substantial pulmonary hypertension, a large intra-alveolar protein leak, peripheral neutropenia, and pathological features of marked neutrophil infiltration with extensive hyaline membrane formation. Pretreatment with indomethacin abolishes postlavage pulmonary hypertension, preserves a slightly better lung function with higher arterial PO2, and prevents the postlavage peripheral neutropenia found in untreated animals. Pretreatment with a thromboxane A2 receptor blocker (L 655,240, Merck Frosst, Canada) also completely attenuated pulmonary hypertension, providing evidence that thromboxane A2 mediates pulmonary arterial hypertension after lung lavage. However, specific thromboxane receptor blockade had no other long-lasting beneficial effects on the ongoing injury in this model.

Sequential pulmonary function measurements during treatment of infantile chronic interstitial pneumonitis.

Three infants with histologically confirmed chronic interstitial pneumonitis were treated with monthly intravenously administered high doses of methylprednisolone with or without daily hydroxychloroquine therapy. We applied the multiple occlusion technique to measure the static respiratory system compliance, and the end-inspiratory occlusion technique to measure passive respiratory system compliance, resistance, and time constant. When assessed by clinical criteria and pulmonary function measurements, all three patients showed improvement with this treatment. Clinical improvement was associated with an increase in respiratory system compliance as measured by both techniques (60% to 100% increase in all patients). The passive respiratory resistance and the time constant did not closely reflect the clinical course. We conclude (1) that high doses (pulses) of methylprednisolone and daily oral doses of hydroxychloroquine are effective in the treatment of infantile chronic interstitial pneumonitis and (2) that the respiratory system compliance, measured by both pulmonary function techniques, correlates well with the response to treatment and change in clinical status.

A model of the maturation of respiratory control in the newborn infant.

A model of automatic neonatal respiratory control has been constructed as an aid in the investigation of a possible maturation in respiratory control loops during the newborn period. The primary objective was to provide a framework for investigating this hypothesis without the need for external stimuli or invasive measurements. Spontaneous sighs provide a physiological disturbance to the respiratory system by transiently altering the levels of the blood gases. The dynamic ventilatory response following such a disturbance was modeled. A change from a highly damped to less damped pattern was found when model parameter values were varied to mimic maturation in the neonatal period. A perturbation model analysis demonstrated the dynamic ventilatory response is most sensitive to factors affecting the gain of the peripheral chemoreflex loop. It is concluded that the model provides valuable insight into the hypothesis that the peripheral chemoreflex matures during the neonatal period and provides a viable method for testing this in the human infant.

Incidências de apnéias obstrutivas em RNS pré-termos / Prevalence of obstructive apneas in the pre-term newborn infant

Estudamos 20 recém-nascidos pré-termos com o objetivo de determinar a incidência de apnéias centrais, mistas e obstrutivas. O peso médio nesta populaçäo foi de 1086 + ou - 211g e a idade na época do estudo foi de 19.1 + ou - 94 dias. Neste grupo, 12 RNS eram portadores de hemorragia intracraniana (HIC) diagnosticada pelo Ultrassom ou Tomografia Computadorizada. Registramos os movimentos respiratórios com eletromiograma e magnetômetros torácicos e o fluxo nasal com um termistor. Observamos um total de 130 apnéias. No grupo sem hemorragia intracraniana houve predominância de episódios do tipo obstrutivo e misto. Concluimos que os episódios do tipo central säo os mais freqüentemente observados em RSN pré-termo e que episódios mistos e obstrutivos predominam somente em recém-nascidos com evidência de hemorragia intracraniana