Electrolytic drops in an electric field: A numerical study of drop deformation and breakup.

The deformation and breakup of an axisymmetric, conducting drop suspended in a nonconducting medium and subjected to an external electric field is numerically investigated here using an electrokinetic model. This model uses a combined level set-volume of fluid formulation of the deformable surfaces, along with a multiphase implementation of the Nernst-Planck equation for transport of ions, that allows for varying conductivity inside the drop. A phase diagram, based on a parametric study, is used to characterize the stability conditions. Stable drops with lower ion concentration are characterized by longer drop shapes than those achieved at higher ion concentrations. For higher drop ion concentration, greater charge accumulation is observed at drop tips. Consequently, such drops break up by pinching off rather than tip streaming. The charge contained in droplets released from unstable drops is shown to increase with drop ion concentration. These dynamic drop behaviors depend on the strength of the electric field and the concentration of ions in the drop and result from the interplay between the electric forces arising from the permittivity jump at the drop interface and the ions in the bulk.

Electroviscous resistance of nanofluidic bends.

Analysis tools that quantify the pressure and potential changes occurring over pressure-driven electrokinetic device elements are necessary for the design of optimal laboratory-on-a-chip devices. In this study, the resistance of a nanofluidic silica channel with negatively charged walls containing a 90^{∘} bend to the electroviscous flow of a potassium chloride salt solution is quantified in terms of two equivalent lengths using numerical analysis. One equivalent length is based on the excess pressure drop and the other on the excess potential rise. Over the entire range of simulations conducted, these equivalent lengths are relatively independent of salt concentration, flow velocity, channel size, and surface charge, remaining within the approximate ranges of 1.3-1.5 for the pressure equivalent length and 0.8-1.05 for the potential equivalent length.

Effect of wall permittivity on electroviscous flow through a contraction.

The electroviscous flow at low Reynolds number through a two-dimensional slit contraction with electric double-layer overlap is investigated numerically for cases where the permittivity of the wall material is significant in comparison with the permittivity of the liquid. The liquid-solid interface is assumed to have uniform surface-charge density. It is demonstrated that a finite wall permittivity has a marked effect on the distribution of ions in and around the contraction, with a significant build-up of counter-ions observed at the back-step. The development length of the flow increases substantially as the wall permittivity becomes significant, meaning that the electric double-layers require a longer distance to develop within the contraction. Consequently, there is a corresponding decrease in the hydrodynamic and electro-potential resistance caused by the contraction. The effect of wall-region width on the flow characteristics is also quantified, demonstrating that the development length increases with increasing wall-region width for widths up to 5 channel widths.

Exploring the native human antibody repertoire to create antiviral therapeutics.

Native human antibodies are defined as those that arise naturally as the result of the functioning of an intact human immune system. The utility of native antibodies for the treatment of human viral diseases has been established through experience with hyperimmune human globulins. Native antibodies, as a class, differ in some respects from those obtained by recombinant library methods (phage or transgenic mouse) and possess distinct properties that may make them ideal therapeutics for human viral diseases. Methods for cloning native human antibodies have been beset by technical problems, yet many antibodies specific for viral antigens have been cloned. In the present review, we discuss native human antibodies and ongoing improvements in cloning methods that should facilitate the creation of novel, potent antiviral therapeutics obtained from the native human antibody repertoire.

Analysis of the original antigenic sin antibody response to the major outer membrane protein of Chlamydia trachomatis.

The anamnestic antibody response to the Chlamydia trachomatis major outer membrane protein (MOMP) was evaluated in mice after priming with serovar C and boosting either with the homologous serovar or with heterologous serovars (A, H, K, and B). Microimmunofluorescence antibody responses demonstrated that boosting with heterologous serovars strongly recalled antibody to serovar C, typical of an original antigenic sin (OAS) response. Boosting with serovars antigenically related to serovar C (A, H, and K) recalled antibody to the variable domain 1 (VD1) peptide of the MOMP of serovar C as determined by a pin-peptide ELISA. Complete amino acid substitution analysis of the VD1 peptide epitope of the MOMP showed that the original antigenic sin response to each boosting serovar contained antibodies with unique patterns of VD1 peptide recognition. The data suggest that antigenically related C. trachomatis serovars differentially recruit B cell lineages from a heterogeneous memory B cell pool that had been induced by priming with the original serovar and thus account for the OAS antibody response.

Mapping epitopes of neutralizing monoclonal antibodies using phage random peptide libraries.

Identification of protective determinants from microbial proteins is a necessary step in the rational design of subunit vaccines. We have previously used a synthetic peptide scan (Pepscan) assay to map a panel of eight neutralizing monoclonal antibodies (mAb; designated as C1.1 to C1.8) to a common motif sequence from Chlamydia trachomatis. In the present study, five of the eight mAbs were used to screen phage random peptide libraries. mAbs C1.1 and C1.3 selected a motif sequence of G-L-X-N-D from a pIII-based phage random peptide library and a motif sequence of G-X-X-N-D from a pVIII-based random peptide library while mAbs C1.6 to C1.8 failed to select recognizable motifs from either of the phage libraries. However, C1.6 to C1.8 bound to the same motif sequence displayed on phage when the appropriate conformational constraints were imposed onto the motif sequence. Thus the specificity of the mAbs identified on Pepscan assays correlates with the mAbs' dependence on local epitope constraints displayed on the phage surface.

Chlamydia trachomatis CTP synthetase: molecular characterization and developmental regulation of expression.

Chlamydia trachomatis is a nucleotide parasite, being entirely dependent on its host eukaryotic cell for a supply of ATP, GTP, and UTP. Chlamydiae are not, however, auxotrophic for CTP, as they are able both to transport CTP from the host and synthesize CTP de novo via a chlamydial CTP synthetase. This study addresses the developmental regulation of CTP synthetase over the course of the C. trachomatis life cycle. Given the distinct life stages of C. trachomatis, analysis of temporal changes in gene expression and regulation of protein activity is the key to unravelling the mechanism of pathogenesis of this bacterium. The results of immunodetection analysis indicate that CTP synthetase is present in C. trachomatis elementary bodies and reticulate bodies and that it is widespread in other chlamydial strains. Reverse transcriptase-polymerase chain reaction (RT-PCR) and metabolic labelling experiments show that CTP synthetase is transcribed and translated primarily during the mid- and late stages of the chlamydial growth cycle. In addition, C. trachomatis CTP synthetase was transcribed with the CTP utilizing enzyme CMP-2-keto-3-deoxy-octanoic acid synthetase (CMP-KDO synthetase) as part of a polycistronic mRNA. The co-expression of these two enzymes suggests a role for CTP synthetase in lipopolysaccharide biosynthesis, potentially channelling CTP directly to CMP-KDO synthetase. The ability of the intact operon to complement CTP synthetase and CMP-KDO deficiencies in mutant Escherichia coli strains indicates that both enzymes are efficiently translated from a single messenger RNA. Kinetic analysis revealed that the C. trachomatis CTP synthetase possessed co-operativity patterns typical of both prokaryotic and eukaryotic CTP synthetases. However, the K(m) of the enzyme for UTP was lower than that of E. coli CTP synthetase, presumably in response to the low intracellular concentration of this nucleotide in C. trachomatis.

Functional and morphologic changes caused by acute ozone exposure in the isolated and perfused rat lung.

Ozone has been shown to increase airway resistance and/or airway reactivity in vivo in animals and humans. Because of the complexities inherent in studying this phenomenon in whole animals, we developed a model of ozone-induced effects on airway physiology using the isolated perfused rat lung. Rat lungs were suspended in an airtight chamber and perfused via the pulmonary circulation with a modified Krebs-Henseleit buffer containing 4.5% bovine albumin. Ventilation of the lungs was achieved by generating a fluctuating negative pressure within the chamber (-2 to -7 cm H2O) at a rate of 60 breaths/min. The lungs were ventilated with humidified 95% air and 5% CO2 alone (control condition) or mixed with ozone at 1.0 or 2.0 ppm. Transpulmonary pressure, flow rate, and tidal volume were recorded at 0, 1, 2, and 3 hours, and pulmonary resistance (RL) and dynamic compliance (Cdyn) were calculated. There was no significant difference in lung weight/total body weight ratios between the three groups at the end of the 3-h period. RL increased and Cdyn decreased in a time- and dose-dependent manner with ozone exposure. The percent increase above baseline in RL +/- SEM at 3 h was 9.4 +/- 4.1% for control lungs, 21.0 +/- 3.2% for 1.0 ppm ozone-exposed lungs, and 63.6 +/- 13.5% for 2.0 ppm ozone-exposed lungs. The percent decrease below baseline in Cdyn +/- SEM at 3 h was 27.4 +/- 2.1% for control lungs, 37.1 +/- 2.7% for 1.0 ppm ozone-exposed lungs, and 55.2 +/- 7.3% for 2.0 ppm ozone-exposed lungs.(ABSTRACT TRUNCATED AT 250 WORDS)

Dynamic respiratory mechanics in monkeys measured by forced oscillations.

We measured impedances of the lungs (ZL) and respiratory system (Zrs) by discrete frequency (f) forced oscillations in adult bonnet monkeys at two lung volumes, FRC and at transrespiratory system pressure (Prs) of -5 cm H2O. Measurements were made from 2 to 32 Hz to study f dependence of effective resistances (Reff) and reactances, and from 1 to 8 Hz to study f dependence of effective pulmonary compliance (Ceff,L). Estimates of resistances (R), inertances (I) and compliances (C) of the lungs (L), chest wall (W) and respiratory system (rs) were obtained by fitting the 2-32 Hz data to a series RIC network model. Reff,L, Reff,W and Reff,rs were found to be f dependent. The magnitude of ZL was small relative to that of Zrs at all f with RL only 30% of Rrs and CL six times Crs. Decreasing lung volume from FRC to Prs = -5 cm H2O increased RL, RW and Rrs, slightly decreased CL, CW and Crs, slightly increased resonance frequencies, and increased the f dependent behavior of Ceff,L. Our results indicate both similarities and differences between the respiratory system of these primates and man.

Respiratory mechanics in normal bonnet and rhesus monkeys.

We measured lung volumes and quasi-static volume-pressure relationships in 22 normal upright bonnet (Macaca radiata) and 12 rhesus (M. mulatta) monkeys. In comparison with interspecies pulmonary function/body weight regressions our monkeys' lung volumes are larger and their lungs are considerably more compliant, but their chest wall compliance is similar to a wide range of mammalian species. However, chest wall compliance of our monkeys was found to be considerably less than that of other more commonly used experimental mammals such as dogs, cats, and rodents. The monkey chest walls were found to be about four times as stiff (3.3 +/- 0.1 (ml/cmH2O)/kg), whereas their lungs were nearly twice as compliant (9.2 +/- 0.7 (ml/cmH2O)/kg) compared to those of supine beagle dogs. Thus, their stiff chest wall sets their functional residual capacity (64.1 +/- 1.2% TLC30) at a much larger percentage of total lung capacity (TLC30) than that of the supine beagle dog (33.8% TLC30). Residual volume (13.2 +/- 1.9% TLC30) equaled the trapped gas volume after bilateral thoracotomy and was set by airway closure. We found more hysteresis area in the chest wall than in the lungs. Our measurements indicate that the static mechanical behavior of the respiratory system of the monkey compares well to man and that the monkey has considerable merit as an animal model for human respiratory function and disease research.

Flow-volume curves and total pulmonary resistance in normal bonnet and rhesus monkeys.

We measured expiratory flow-volume curves and total pulmonary resistance in 15 normal, anesthetized, upright bonnet (Macaca radiata) and 8 rhesus (M. mulatta) monkeys. Absolute maximum flows are about half that found in humans, but, if expressed in vital capacities per second, monkey flows are about four times that of humans. However, irrespective of flow rate, the typical shape of maximum expiratory flow-volume (MEFV) curves of rhesus monkeys is similar to that of normal young adults and that of bonnet monkeys is similar to that of normal older adults. Flow-limiting mechanisms that may determine MEFV curve shape and explain the differences and similarities of monkey and human MEFV curves are discussed. We propose that differences in function between bonnet and rhesus monkeys relates to the concept of dysanaptic lung growth that evolved to subserve behavioral differences between the two species of monkey.

Influence of ruminal insufflation on pulmonary function and diaphragmatic electromyography in cattle.

In 8 healthy, awake cows with permanent cannulated ruminal fistulas and carotid artery loops, respiratory mechanics, ventilation, and diaphragmatic electrical activity were studied before and during stepwise insufflation of the rumen with air pressure to 40 mm of Hg. We found that ruminal insufflation increased intraperitoneal, intrapleural, and transdiaphragmatic pressures and decreased lung volume and lung compliance. In individual cows with rumen insufflation there was an increase in pulmonary resistance, but this trend was not significant in the group. Peak expiratory flow rate was increased and peak inspiratory flow rate was unchanged. Inspiratory duration (Ti) decidal volume decreased slightly, breathing frequency decreased markedly, and minute volume decreased. When intraruminal pressure reached 40 mm of Hg, arterial partial pressure of carbon dioxide (PaCO2) increased (P less than 0.01) and that of oxygen (PaO2) decreased (P less than 0,01) and arterial blood pH decreased (P less than 0.02). Diaphragmatic electromyographic activity was increased, but mechanical effectiveness of the diaphragm was reduced at increased intraruminal pressures.

Flow rates from an intermittent positive pressure breathing-anesthetic delivery apparatus for horses.

Inspiratory flow rates were measured and compared in an equine intermittent positive pressure breathing-anesthesia delivery apparatus powered by a positive phase ventilator, a positive-negative phase ventilator, and a modified positive phase ventilator with automatic flow acceleration at driving pressures of 2,600, 4,400, and 5,200 mm of Hg (50, 85, and 100 psi). The last-named apparatus consistently produced the highest flows for a given ventilator setting and driving pressure. Regardless of the unit used, the greater driving pressures and high ventilator range settings produced the greatest flows.

Effect of halothane and halothane-nitrous oxide on hematocrit and plasma protein concentration in dog and monkey.

Hematocrit and plasma protein concentration in healthy dogs and monkeys (Macaca arctoides) awake and anesthetized with halothane-oxygen and halothane-nitrous oxide oxygen were compared during conditions of spontaneous and controlled ventilation. Both hematocrit and plasma protein concentration decreased within 15 minutes following anesthetic induction. This decrease persisted throughout constant- or variable-depth anesthesia and did not vary appreciably with ventilation, anesthetic dose, or introduction of nitrous oxide. Plasma volume, determined by a dye dilution technique, concomitantly increased. This increase is compatible with the directional changes in hematocrit and plasma protein.

Circulatory effects of halothane and halothane-nitrous oxide anesthesia in the dog: spontaneous ventilation.

The cardiovascular effects of equipotent (minimum alveolar concentration; MAC) doses of halothane versus halothane plus 25% N2O (H25N2O) in spontaneously breathing dogs do not differe except that nitrous oxide increased mean arterial pressure (AP) and decreased arterial oxygen partial pressure (PAO2). When 75% nitrous oxide was added to halothane anesthesia, AP, mean pulmonary artery pressure (PAP), heart rate (HR), cardiac output (CO), stroke volume (SV), total peripheral resistance (TPR), and left ventricular work (LVW) increased and PAO2 and hemoglobin saturation decreased. Arterial oxygen tensions below 80 torr were common at moderate and deep anesthetic levels of halothane plus 75% N2O (H75N2O). The specific contribution of N2O, hypoxemia, hypercapnia, or temporal recovery (or a combination of these) in producing cardiovascular stimulation were not determined.