Transient oscillatory force-length behavior of activated airway smooth muscle.

Airway smooth muscle (ASM) is cyclically stretched during breathing, even in the active state, yet the factors determining its dynamic force-length behavior remain incompletely understood. We developed a model of the activated ASM strip and compared its behavior to that observed in strips of rat trachealis muscle stimulated with methacholine. The model consists of a nonlinear viscoelastic element (Kelvin body) in series with a force generator obeying the Hill force-velocity relationship. Isometric force in the model is proportional to the number of bound crossbridges, the attachment of which follows first-order kinetics. Crossbridges detach at a rate proportional to the rate of change of muscle length. The model accurately accounts for the experimentally observed transient and steady-state oscillatory force-length behavior of both passive and activated ASM. However, the model does not predict the sustained decrement in isometric force seen when activated strips of ASM are subjected briefly to large stretches. We speculate that this force decrement reflects some mechanism unrelated to the cycling of crossbridges, and which may be involved in the reversal of bronchoconstriction induced by a deep inflation of the lungs in vivo.

A further analysis of why pulmonary venous pressure rises after the onset of LV dysfunction.

Based on a dynamic computational model of the circulation, Burkhoff and Tyberg (Am J Physiol Heart Circ Physiol 265: H1819-H1828, 1993) concluded that the rise in pulmonary venous pressure (Pvp) with left ventricular (LV) dysfunction requires a decrease in vascular capacitance and transfer of unstressed volume to stressed volume (nu). We argue that the values they used for venous resistance (Rvs), venous compliance (Cvs), and nu were too low, and changing these values significantly changes the conclusion. We used a computational model of the circulation that was similar to theirs, but we made Rvs four times higher (0.06 versus 0.015 mmHg.s.ml(-1)), Cvs larger (110 versus 70 ml/mmHg), and nu larger (1,400 versus 750 ml); all other parameters, including those for the heart, were essentially the same. We simulated left ventricular dysfunction by decreasing end-systolic elastance (Eeslv) as they did and examined changes in cardiac output, arterial blood pressure, and Pvp. We then examined the effect of changes in Rvs, heart rate, and nu when Eeslv was depressed with and without pericardial constraint. In contrast to their findings, with our parameters the model predicts that decreasing Eeslv substantially increases Pvp. Furthermore, increasing systemic vascular resistance or decreasing Rvs or heart rate produces large increases in Pvp when Eeslv is reduced. Pericardial constraint limits the changes in Pvp. In conclusion, when Rvs and Cvs are increased, baseline nu must be higher to maintain normal cardiac output. This increased volume can shift between compartments under flow conditions and account for the increase in Pvp with decreased left ventricular function even without recruitment of unstressed volume.

Increasing likelihood of advanced pulmonary tuberculosis at initial diagnosis in a low-incidence US state.

SETTING: Arkansas, USA. OBJECTIVE: To investigate the relationship between an increase in the proportion of cases with advanced disease at first diagnosis and the recently observed slowing of the decline in tuberculosis (TB) incidence in low-incidence US states. DESIGN: We conducted descriptive statistical analyses of de-identified surveillance data of 1246 culture-confirmed TB patients reported in Arkansas during 1996-2013. We then fitted stepwise, multivariate logistic regression models to identify predictors for advanced disease at diagnosis, defined as having either smear-positive sputum or lung cavitation. RESULTS: From 1996 to 2013, the proportion of new cases with positive sputum smear and cases with lung cavitation increased from 51.6% to 75% and from 37.7% to 50%, respectively. Patients diagnosed during 2006-2013 were more likely to have positive sputum smears (adjusted odds ratio [aOR] 2.55, 95%CI 1.95-3.35) or lung cavitation (aOR 1.49, 95%CI 1.14-1.95) than those diagnosed during 1996-2005. During 1996-2013, age 15-64 years and excessive alcohol use were predictive of positive sputum smear or lung cavitation. CONCLUSION: Measures to reduce the proportion of cases with advanced disease at first diagnosis may be helpful to achieve further decline in TB incidence in low-incidence settings.

Effects of thermal adaptation at 40 degrees C on membrane viscosity and the sodium-potassium pump in Chinese hamster ovary cells.

We have demonstrated increased heat resistance in Chinese hamster ovary cells grown to confluence at 37 degrees C and thermally adapted at the nonlethal temperature of 40 degrees C for 24 h. Membrane viscosity, estimated by fluorescence anisotropy, was inversely related to temperature, from 5 degrees C to 45 degrees C. For a given temperature viscosity was consistently higher in thermally adapted cells than in native cells. Having demonstrated a change in membrane structure associated with thermal adaptation, we carried out a study of the Na+-K+ pump in native and thermally adapted cells as an example of a vital active transport process known to be sensitive to membrane viscosity. 86Rb uptake measured from 31 degrees C to 50 degrees C increased steadily to 46 degrees C and then decreased rapidly in both native and thermally adapted cells. Detailed measurement of ouabain-sensitive 86Rb influx demonstrated an increase in both Km and Vmax between 37 degrees C and 45 degrees C, but there was no difference between native and thermally adapted cells. We have thus demonstrated an adaptive structural change in the cell membrane of mammalian cells which may be related to the induction of thermal resistance at 40 degrees C but which is not associated with any change in this active transport system.

Thermoregulatory failure secondary to acute illness: complications and treatment.

We report on six patients in whom hypothermia secondary to acute illnesses, including pneumonia, congestive heart failure, renal failure, drug overdose, and hypoglycemia, developed. Complications that occurred were metabolic acidosis in six patients, altered sensorium in five, bradyarrhythmia in three, and hyperamylasemia in two. All patients failed to demonstrate a shivering response and represent cases of acute thermoregulatory failure. Five of the six patients survived. In the course of treatment, the choice of active or passive rewarming should be based on whether or not normal thermoregulatory mechanisms are intact.

Restriction endonuclease mapping and cloning of Mycobacterium intracellulare plasmid pLR7.

A restriction map of Mycobacterium intracellulare plasmid pLR7 was developed. This 15.3-kb plasmid had unique sites for BamHI, HindIII, and XbaI. Various large fragments of pLR7 were cloned into pBR322 or pHP34 and propagated in Escherichia coli. A hybrid pLR7 ::pBR322 plasmid carrying the complete pLR7 sequence was constructed by joining the plasmids at their HindIII sites. The construction of these hybrids will facilitate the analysis and manipulation of pLR7 and may allow the development of this plasmid as a model system for genetic analysis in mycobacteria.

Effects of cholecystokinin tetrapeptide on respiratory function in healthy volunteers.

OBJECTIVE: The authors evaluated respiratory response to cholecystokinin tetrapeptide (CCK-4) in healthy volunteers. METHOD: Subjects were randomly assigned to either a CCK-4 (N = 15) or placebo (N = 15) challenge under double-blind conditions. RESULTS: Dyspnea was reported by all of the subjects who received CCK-4 but only one subject who received placebo. CCK-4 caused a significant increase in tidal volume and minute ventilation but had no effect on breathing frequency. Placebo had no effect on any of the respiratory measures. CONCLUSIONS: These data indicate that the behavioral effects of CCK-4 are accompanied by changes in respiration in healthy volunteers.

The effect of delay in treatment on local control by radiotherapy.

PURPOSE: The objective of this study was to estimate the effect of delay in initiation of treatment on rates of local control by radiotherapy. METHODS AND MATERIALS: A model of the effects of delay was developed based on the following assumptions: (a) that tumor growth rate is exponential, (b) that a predetermined radiotherapy regimen will kill the same fraction of clonogenic cells in a given tumor whether it is administered early or late, and (c) that the absolute number of cells surviving in a tumor is determined by Poisson statistics. Monte Carlo simulation was used to estimate the expected rate of decrease in local control associated with delay in a population of tumors, which was heterogeneous with respect to doubling time and initial volume. The model was applied to carcinoma of the tonsillar region. RESULTS: It was shown that at some point in the evolution of every case, the probability of local control decreases sharply over a relatively short period of time. The maximum rate of decrease in the probability of local control occurs at the 37% local control level when it reaches 25.5% per tumor doubling time. When heterogeneity with respect to doubling time and stage was taken into account, it was estimated that the local control rate would decrease by approximately 10% per month in a typical series of patients with carcinoma of the tonsillar region. CONCLUSIONS: It was concluded that delay in initiation of radiotherapy may be associated with a clinically important deterioration in local control rates. We recommend that waiting times for radiotherapy should be As Short As Reasonably Achievable (ASARA).

Temperature dependence of melphalan efflux kinetics in Chinese hamster ovary cells.

The temperature dependence of the kinetics of efflux of melphalan from Chinese hamster ovary (CHO) cells was studied from 4 degrees to 47 degrees. Time courses for melphalan efflux showed an initial rapid phase of efflux followed by a plateau. The data for melphalan concentration (c) versus efflux time (t) were described by the equation c(t) = A + B exp(-kt), where A is the final steady-state melphalan concentration, B is the total change in melphalan concentration from time zero until steady-state conditions are reached, and k is the rate constant for the efflux process. The plateau level obtained was not dependent on temperature and corresponded to 22 +/- 3.2% of the drug remaining in the cells after efflux. The time for melphalan efflux to reach the plateau level was dependent on temperature. This was reflected by an increase in the rate constants for melphalan efflux with increasing temperature from 30 degrees to 47 degrees. The rate constant for melphalan efflux at 37 degrees was 0.045 +/- 0.002 min-1. Efflux of melphalan occurred much more slowly at lower temperatures such as 4 degrees and 20 degrees. An Arrhenius plot for melphalan efflux showed a linear and decreasing trend at temperatures between 30 degrees and 47 degrees with an activation energy of 1.046 x 10(3) J/mol.

Rapid definitive diagnosis of Legionnaires' disease.

We describe a sporadic case of Legionnaires' disease in which the diagnosis was made by direct immunofluorescence of material obtained by percutaneous aspiration of the involved lung via a needle. Employment of this technique among selected patients with suspected Legionnaires' disease would provide for more rapid diagnosis and more prompt initiation of definitive therapy for some patients.

Microbial etiology of acute pneumonia in hospitalized patients.

The purpose of this study was to determine the microbial etiology of pneumonia by using strict criteria among a group of hospitalized patients. Patients with acute community-acquired or hospital-acquired pneumonia were studied in a systematic and comprehensive manner for bacterial, viral, chlamydial, mycobacterial, and fungal pathogens. A total of 198 patients with 204 episodes of pneumonia were evaluated. Despite 100 percent follow-up of all surviving patients, a specific etiologic agent could be found in only 103 episodes. Among 154 episodes of community-acquired pneumonia, a diagnosis was made in 79; the most common pathogen was from the genus Legionella, followed by various Gram-negative enteric bacteria, Gram-positive cocci, influenza A virus, and Mycoplasma pneumoniae. The etiologic agent was found in 24 of the 50 patients with hospital-acquired pneumonia; no pathogen predominated. We conclude that even when elaborate diagnostic studies are done, including many invasive procedures, the etiology can be determined in only about half of the patients with acute pneumonia. The pathogens of pneumonia in this study are not markedly different between community-acquired and hospital-acquired infection.

A model-free way of representing hyperthermia cell survival data.

We present a method of fitting curves to cell survival data that is free from all model assumptions, requiring only that the fitted curves be decreasing and reasonably smooth, where the degree of smoothness is determined from considerations of experimental error. The fitted curves are then differentiated to yield frequency distributions of cell killing times, which may be of value in defining subpopulations with different sensitivities to the cytotoxic agent under study. In addition, confidence intervals on the fitted curves and frequency distributions are obtained by Monte Carlo simulation. The results allow the objective and model-free assessment of the effects of various experimental interventions on cell survival.

Thermal adaptation in CHO cells at 40 degrees C: the influence of growth conditions and the role of heat shock proteins.

The kinetics of thermal adaptation at the nonlethal temperature of 40 degrees C was studied in CHO (Chinese hamster ovary) cells in vitro. Thermal resistance, demonstrated as an increase in mean 45 degrees C killing time or as an increase in the shoulder of the 45 degrees C survival curve, was fully developed by 2 h. Control cells in early logarithmic phase were more heat sensitive than those in stationary phase. Corresponding 45 degrees C killing time frequency distributions were unimodal with an increase in mean killing time from early logarithmic to stationary phase. Cells which were thermally adapted at 40 degrees C for 6 h had biphasic 45 degrees C killing time frequency distributions, and as cells progressed from early logarithmic to stationary phase the heat-sensitive subpopulation progressively declined. Exposure to 40 degrees C produced a 30% increase in total protein synthesis. Proteins with molecular weights 72, 89, and 109 kDa which correspond to those induced by lethal heat shock were synthesized at 40 degrees C, but there was no close temporal correlation between the development of heat resistance at 40 degrees C and synthesis of the heat shock proteins. Cycloheximide (100 micrograms/ml) reduced the mean 45 degrees C killing time but did not totally prevent the development of heat resistance at 40 degrees C.

Ventilatory response to cholecystokinin tetrapeptide in anaesthetized dogs.

Cholecystokinin (CCK) has been implicated in the genesis of panic disorder. In this study we measured the ventilatory response to i.v. injection of CCK-tetrapeptide (CCK-4) in anaesthetized dogs. We found an immediate and transient increase in minute ventilation. The response was large (more than 100% of baseline) and lasted 1-2 min. Repeated CCK-4 injection produced a somewhat reduced response, suggestive of a slight degree of tachyphylaxis. Blood pressure and heart rate changed only slightly (<10%), while respiratory mechanical parameters remained unchanged following CCK-4 administration. We conclude that CCK-4 has a significant effect on ventilation in the anaesthetized dog, which suggests that this species may provide a useful quantitative assay for the anxiogenic effects of CCK.

Stochastic model of the pulmonary airway tree and its implications for bronchial responsiveness.

The resistance of the pulmonary conducting airway tree (Raw) is a consequence of the resistances of its component airways and how they are connected together. To date, theoretical calculations of Raw have been performed with the aid of mathematical models of the airway tree that are purely deterministic. That is, the mechanical properties of the component airways in these models are precisely defined functions of generation number. Such models take no account of the fact that the airways of a given generation are not all exactly the same but rather exhibit a spectrum of wall thicknesses, amounts of smooth muscle, and number of parenchymal attachments. In the present study, the properties of a 10-generation stochastic airway tree model are investigated. The lengths and radii of the airways in the tree are drawn randomly from probability distribution functions (PDFs), the means of which are deterministic functions of generation number and the standard deviations are assigned various values. Monte Carlo simulation is used to estimate the PDF of Raw itself in various conditions. We show that the relative width of the PDF of Raw may be comparable to that of the PDFs from which the individual airway radii were drawn. It is also shown that when bronchoconstriction is simulated by narrowing each airway by a random amount the resulting PDF for Raw may increase in width many times. We conclude that the variations in airway responsiveness seen in nature can only be properly understood when the distribution of airway properties within the lung are taken into account.

Acute pulmonary response to intravenous histamine at fixed lung volume in dogs.

We measured tracheal pressure (Ptr), tracheal flow, and two alveolar pressures in five open-chest anesthetized and paralyzed dogs. The lungs were maintained at a fixed volume for 50 s while small amplitude oscillations in flow at 6 Hz were applied at the tracheal opening. The measurements of alveolar pressure showed that the resulting oscillations in Ptr were virtually entirely determined by airway resistance (Raw) and consequently gave accurate estimates of the same. A 20-mg bolus of histamine was given intravenously at the start of this period when Ptr was 0.5 kPa. After approximately 10 s the mean Ptr increased sharply by approximately 40% and plateaued after approximately 25 s. Raw, in contrast, continued to increase throughout the oscillation period. Furthermore, the increases in mean Ptr were virtually identical in all dogs, whereas the increases in Raw were highly variable among the dogs. Our results suggest that the increases in mean Ptr caused by histamine were due to contraction of distal elements in the lung, whereas the changes in Raw were due mainly to constriction of more central airways.

Effect of stochastic heterogeneity on lung impedance during acute bronchoconstriction: a model analysis.

In a previous study (J. H. T. Bates, A. M. Lauzon, G. S. Dechman, G. N. Makaym, and T. F. Schuessler. J. Appl. Physiol. 76: 616-626, 1994), we investigated the acute changes in isovolume lung mechanics immediately after a bolus injection of histamine. We found that dynamic resistance and elastance increased progressively in the 80-s period after injection, whereas the estimated tissue hysteresivity reached a stable plateau after approximately 25 s. In the present study, we developed a computer model of the lung to investigate the mechanisms responsible for these observations. The model conforms to Horsfield's morphometry, with the addition of compliant airways and structural damping tissue units. Using this model, we simulated the time course of acute bronchoconstriction after intravenous administration of a bolus of bronchial agonist. Heterogeneity was induced by randomly varying the values of the maximal airway smooth muscle contraction and the tissue response to the agonist. Our results demonstrate that much of the increase in lung impedance observed in our previous study can be produced purely by the effects of airway heterogeneity. However, we were only able to reproduce the plateauing of hysteresivity by assigning a minimum radius to each airway, beyond which it would immediately snap completely shut. We propose that airway closure played an important role in our experimental observations.

Breathing responses to small inspiratory threshold loads in humans.

To investiage the effect of inspiratory threshold load (ITL) on breathing, all previous work studied loads that were much greater than would be encountered under pathophysiological conditions. We hypothesized that mild ITL from 2.5 to 20 cmH2O is sufficient to modify control and sensation of breathing. The study was performed in healthy subjects. The results demonstrated that with mild ITL 1) inspiratory difficulty sensation could be perceived at an ITL of 2.5 cmH2O; 2) tidal volume increased without change in breathing frequency, resulting in hyperpnea; and 3) although additional time was required for inspiratory pressure to attain the threshold before inspiratory flow was initiated, the total inspiratory muscle contraction time remained constant. This resulted in shortening of the available time for inspiratory flow, so that the tidal volume was maintained or increased by significant increase in mean inspiratory flow. On the basis of computer simulation, we conclude that the mild ITL is sufficient to increase breathing sensation and alter breathing control, presumably aiming at maintaining a certain level of ventilation but minimizing the energy consumption of the inspiratory muscles.

Estimation of time-varying respiratory mechanical parameters by recursive least squares.

Continuous estimation of time-varying respiratory mechanical parameters is required to fully characterize the time course of bronchoconstriction. To achieve such estimation, we developed an estimator that uses the recursive linear least-squares algorithm to fit the equation Ptr = RV + EV + K to measurements of tracheal pressure (Ptr) and flow (V). The volume (V) is obtained by numerical integration of V. The estimator has a finite memory with length into the past at each point in time that varies inversely with the difference between the current measurement of Ptr and that predicted by the model, to allow the algorithm to track rapidly varying parameters (R, E, and K). V usually exhibits significant drift and must be corrected. Of the several correction methods investigated, subtraction of the recursively weighted average of V before integration to V was found to perform best. The estimator was tested on simulated noisy data where it successfully followed a fivefold increase in R and a twofold increase in E occurring over 10 s. Three dogs and two cats were anesthetized, paralyzed, tracheostomized, and challenged with a bolus of methacholine (approximately 13 mg/kg iv). Increases of 3- to 10-fold were observed in R and 2- to 3-fold in E, beginning within 10-40 s after the bolus injection. In some animals we found that the increase in E occurred more slowly than that in R, which the V signal suggested was due to dynamic hyperinflation of the lungs. These results demonstrate that our recursive estimator is able to track rapid changes in respiratory mechanical parameters during bronchoconstrictor challenge.