The use of chilled condensers for the recovery of perfluorocarbon liquid in an experimental model of perfluorocarbon vapour loss during neonatal partial liquid ventilation

Background  

Perfluorocarbon (PFC) vapour in the expired gases during partial liquid ventilation should be prevented from entering the atmosphere and recovered for potential reuse.     

Use of perfluorocarbon in the culture of Saccharomyces cerevisiae

The growth of Saccharomyces cerevisiae was enhanced by using perfluorocarbon (PFC) while it was inhibited by pluronic F-68 alone. The use of 15%(v/v) PFC resulted in almost twice as much biomass concentration compared to the control. The inclusion of PFC emulsion in the culture medium also increased the maximum biomass concentration, but not as much as in the case of pure PFC.     

Enhancement of autotrophic growth rate ofAlcaligenes eutrophus in a medium containing perfluorocarbon under low oxygen partial pressure

Summary Under low oxygen partial pressure,Alcaligenes eutrophus was cultivated autotrophically in a medium containing perfluorocarbon (PFC) as an oxygen-carrier. In the presence of PFC of more than 70% of the medium volume, enhancement of the growth rate was observed. At 90% PFC, the rate was about 18 times greater than that at 0% PFC.     

Effect of the interfacial surfactant layer on oxygen transfer through the oil/water phase boundary in perfluorocarbon emulsions

The effect of interfacial surfactant molecules on oxygen transfer through oil/water phase boundary has been studied in FlurO(2) (TM) emulsions, i.e., perfluorocarbon (PFC) emulsions developed as oxygen carriers in cell culture. Measurements of oxygen permeability were made with a polarographic oxygen electrode in pure PFCs and in emulsions with various PFC volume fractions. Comparison of the experimental results with the theoretically derived values of relative oxygen permeability clearly indicates that the mass transfer resistance caused by the interfacial surfactant layer in PFC emulsions is insignificant. Therefore, oxygen dissolved in the enclosed PFC phase is readily available to cells growing in the aqueous media and FlurO(2) emulsions with very fine emulsion particles (< 0.2 mum) can be used to effectively enhance gas/liquid interfacial oxygen transfer in bioreactors. The inadequacy in describing mass transfer in heterogeneous systems, such as the PFC emulsions, by conventional concentration-based oxygen diffusion coefficients has also been discussed.     

Brain oxygenation and CNS oxygen toxicity after infusion of perfluorocarbon emulsion

Intravenous perfluorocarbon (PFC) emulsions, administered with supplemental inspired O(2), are being evaluated for their ability to eliminate N(2) from blood and tissue prior to submarine escape, but these agents can increase the incidence of central nervous system (CNS) O(2) toxicity, perhaps by enhancing O(2) delivery to the brain. To assess this, we infused a PFC emulsion (Oxycyte, 6 ml/kg iv) into anesthetized rats and measured cerebral Po(2) and regional cerebral blood flow (rCBF) in cortex, hippocampus, hypothalamus, and striatum with 100% O(2) at 1, 3, or 5 atmospheres absolute (ATA). At 1 ATA, brain Po(2) stabilized at >20 mmHg higher in animals infused with PFC emulsion than in control animals infused with saline, and rCBF fell by ~10%. At 3 ATA, PFC emulsion raised brain Po(2) >70 mmHg above control levels, and rCBF decreased by as much as 25%. At 5 ATA, brain Po(2) was ≥159 mmHg above levels in control animals for the first 40 min but then rose sharply; rCBF showed a similar profile, reflecting vasoconstriction followed by hyperemia. Conscious rats were also pretreated with PFC emulsion at 3 or 6 ml/kg iv and exposed to 100% O(2) at 5 ATA. At the lower dose, 80% of the animals experienced seizures by 33 min compared with 50% of the control animals. At the higher dose, seizures occurred in all rats within 25 min. At these doses, administration of PFC emulsion poses a clear risk of CNS O(2) toxicity in conscious rats exposed to hyperbaric O(2) at 5 ATA.     

Shunt and ventilation-perfusion distribution during partial liquid ventilation in healthy piglets

Mates, Elisabeth A., Jacob Hildebrandt, J. Craig Jackson,Peter Tarczy-Hornoch, and Michael P. Hlastala. Shunt and ventilation-perfusion distribution during partial liquid ventilation inhealthy piglets. J. Appl. Physiol.82(3): 933-942, 1997.Replacing gas in the lung withperfluorocarbon fluids (PFC) and periodically ventilating with a gas[partial liquid ventilation (PLV)] has been shown toimprove oxygenation in models of respiratory distress syndrome. Wehypothesized that the addition of PFC to healthy lungs would result inshunt, diffusion impairment, and increased ventilation-perfusion(A/) heterogeneity.Previously, Mates et al. showed thatO₂ shunt and arterial-alveolarCO₂ difference increased linearlywith dose in piglets given graded intratracheal doses of PFC (10, 20, and 30 ml/kg followed by mechanical ventilation with 100%O₂) (E. A. Mates, J. C. Jackson, J. Hildebrandt, W. E. Truog, T. A. Standaert, and M. P. Hlastala. In: Oxygen Transport to TissueXVI, 1994, p. 427-435). Here we reportA/ distribution inthe same animals, showing a 50% increase inA/ heterogeneity during PLV independent of PFC dose. Ventilation heterogeneity was themajor factor in this increase, and there was no significant change indead space ventilation. We also report on five animals given a single20 ml/kg dose of PFC and followed for 3 h. They showed an increase inshunt during PLV but no change in arterial-alveolar CO₂ difference.

     

A remedy to oxygen limitation problem in antibiotic production: addition of perfluorocarbon

In this study, the effect of an oxygen carrier, perfluorocarbon, on actinorhodin fermentation by Streptomyces coelicolor A3(2) was investigated using a chemically defined medium in 2 and 20 l bioreactors. The inclusion of 50% (v/v) perfluorocarbon in the fermentation medium resulted in a five-fold increase in the maximum antibiotic concentration. The use of perfluorocarbon also caused remarkable increases in both glucose and oxygen consumption rates. Moreover, the increasing concentrations of perfluorocarbon improved the dissolved oxygen profile by raising the minimum dissolved oxygen concentration. It was found that observed increases in the antibiotic production were linearly related to the volumetric oxygen uptake rates. This result could perhaps be attributed to the enhancement of oxygen transfer in S. coelicolor cultures due to the higher oxygen solubilities of the fermentation medium through inclusion of perfluorodecalin.     

Alterations of alveolar type II cells and intraalveolar surfactant after bronchoalveolar lavage and perfluorocarbon ventilation. An electron microscopical and stereological study in the rat lung

Background

Repeated bronchoalveolar lavage (BAL) has been used in animals to induce surfactant depletion and to study therapeutical interventions of subsequent respiratory insufficiency. Intratracheal administration of surface active agents such as perfluorocarbons (PFC) can prevent the alveolar collapse in surfactant depleted lungs. However, it is not known how BAL or subsequent PFC administration affect the intracellular and intraalveolar surfactant pool.

Methods

Male wistar rats were surfactant depleted by BAL and treated for 1 hour by conventional mechanical ventilation (Lavaged-Gas, n = 5) or partial liquid ventilation with PF 5080 (Lavaged-PF5080, n = 5). For control, 10 healthy animals with gas (Healthy-Gas, n = 5) or PF5080 filled lungs (Healthy-PF5080, n = 5) were studied. A design-based stereological approach was used for quantification of lung parenchyma and the intracellular and intraalveolar surfactant pool at the light and electron microscopic level.

Results

Compared to Healthy-lungs, Lavaged-animals had more type II cells with lamellar bodies in the process of secretion and freshly secreted lamellar body-like surfactant forms in the alveoli. The fraction of alveolar epithelial surface area covered with surfactant and total intraalveolar surfactant content were significantly smaller in Lavaged-animals. Compared with Gas-filled lungs, both PF5080-groups had a significantly higher total lung volume, but no other differences.

Conclusion

After BAL-induced alveolar surfactant depletion the amount of intracellularly stored surfactant is about half as high as in healthy animals. In lavaged animals short time liquid ventilation with PF5080 did not alter intra- or extracellular surfactant content or subtype composition.     

Visualization of abscess formation in a murine thigh infection model of Staphylococcus aureus by 19F-magnetic resonance imaging (MRI)

Background

During the last years, ¹⁹F-MRI and perfluorocarbon nanoemulsion (PFC) emerged as a powerful contrast agent based MRI methodology to track cells and to visualize inflammation. We applied this new modality to visualize deep tissue abscesses during acute and chronic phase of inflammation caused by Staphylococcus aureus infection.

Methodology and Principal Findings

In this study, a murine thigh infection model was used to induce abscess formation and PFC or CLIO (cross linked ironoxides) was administered during acute or chronic phase of inflammation. 24 h after inoculation, the contrast agent accumulation was imaged at the site of infection by MRI. Measurements revealed a strong accumulation of PFC at the abscess rim at acute and chronic phase of infection. The pattern was similar to CLIO accumulation at chronic phase and formed a hollow sphere around the edema area. Histology revealed strong influx of neutrophils at the site of infection and to a smaller extend macrophages during acute phase and strong influx of macrophages at chronic phase of inflammation.

Conclusion and Significance

We introduce ¹⁹F-MRI in combination with PFC nanoemulsions as a new platform to visualize abscess formation in a murine thigh infection model of S. aureus. The possibility to track immune cells in vivo by this modality offers new opportunities to investigate host immune response, the efficacy of antibacterial therapies and the influence of virulence factors for pathogenesis.     

Modeling diffusion limitation of gas exchange in lungs containing perfluorocarbon

We reportedchanges in alveolar-arterial PO₂ gradient,ventilation-perfusion heterogeneity, and arterial-alveolarPCO₂ gradient during partial liquid ventilation(PLV) in healthy piglets (E. A. Mates, P. Tarczy-Hornoch, J. Hildebrandt, J. C. Jackson, and M. P. Hlastala. In: OxygenTransport to Tissue XVII, edited by C. Ince. New York: Plenum,1996, vol. 388, p. 585-597). Here we develop two mathematicalmodels to predict transient and steady-state (SS) gas exchangeconditions during PLV and to estimate the contribution of diffusionlimitation to SS arterial-alveolar differences. In the simplest model,perfluorocarbon is represented as a uniform flat stirred layer and, ina more complex model, as an unstirred spherical layer in a ventilatedterminal alveolar sac. Time-dependent solutions of both models showthat SS is established for various inert and respiratory gases within5-150 s. In fluid-filled unventilated terminal units, all times toSS increased sometimes by hours, e.g., SF₆ exceeded 4 h. SSsolutions for the ventilated spherical model predicted minorend-capillary disequilibrium of inert gases and significantdisequilibrium of respiratory gases, which could explain a largeportion of the arterial-alveolar PCO₂ gradient measured during PLV (14). We conclude that, during PLV, diffusion gradients for gases are generally small, except for CO₂.

     

Effect of viscosity on instilled perfluorocarbon distribution in rabbit lungs

The effect of viscosity on the distribution of perfluorocarbon instilled into the lungs for liquid ventilation was investigated. Perfluorocarbon (either perfluorodecalin or FC-3283) was instilled into the trachea during ventilation at a constant infusion rate of 40 ml/min and radiographic images were obtained at 30 frames/s. Image analysis was performed and the homogeneity index of the distribution was computed for images at the end of inspiration of each breath to evaluate the evolution of perfluorocarbon distribution during filling. The higher viscosity perfluorocarbon (perfluorodecalin) resulted in a more homogeneous distribution. This was attributed to perfluorodecalin's higher propensity to form liquid plugs in large airways and to those plugs leaving behind a thicker liquid layer as they propagated through the lungs.     

Decreased response inhibition in middle-aged male patients with type 2 diabetes

Background  

This study was performed to examine whether patients with type 2 diabetes have cognitive deficits associated with the prefrontal cortex (PFC).     

In Vitro Surfactant and Perfluorocarbon Aerosol Deposition in a Neonatal Physical Model of the Upper Conducting Airways

Objective

Aerosol delivery holds potential to release surfactant or perfluorocarbon (PFC) to the lungs of neonates with respiratory distress syndrome with minimal airway manipulation. Nevertheless, lung deposition in neonates tends to be very low due to extremely low lung volumes, narrow airways and high respiratory rates. In the present study, the feasibility of enhancing lung deposition by intracorporeal delivery of aerosols was investigated using a physical model of neonatal conducting airways.

Methods

The main characteristics of the surfactant and PFC aerosols produced by a nebulization system, including the distal air pressure and air flow rate, liquid flow rate and mass median aerodynamic diameter (MMAD), were measured at different driving pressures (4–7 bar). Then, a three-dimensional model of the upper conducting airways of a neonate was manufactured by rapid prototyping and a deposition study was conducted.

Results

The nebulization system produced relatively large amounts of aerosol ranging between 0.3±0.0 ml/min for surfactant at a driving pressure of 4 bar, and 2.0±0.1 ml/min for distilled water (H₂Od) at 6 bar, with MMADs between 2.61±0.1 µm for PFD at 7 bar and 10.18±0.4 µm for FC-75 at 6 bar. The deposition study showed that for surfactant and H₂Od aerosols, the highest percentage of the aerosolized mass (∼65%) was collected beyond the third generation of branching in the airway model. The use of this delivery system in combination with continuous positive airway pressure set at 5 cmH₂O only increased total airway pressure by 1.59 cmH₂O at the highest driving pressure (7 bar).

Conclusion

This aerosol generating system has the potential to deliver relatively large amounts of surfactant and PFC beyond the third generation of branching in a neonatal airway model with minimal alteration of pre-set respiratory support.     

Distribution dynamics of perfluorocarbon delivery to the lungs: an intact rabbit model.

Motivated by the goal of understanding how to most homogeneously fill the lungs with perfluorocarbon for liquid ventilation, we investigate the transport of liquid instilled into the lungs using an intact rabbit model. Perfluorocarbon is instilled into the trachea of the ventilated animal. Radiographic images of the perfluorocarbon distribution are obtained at a rate of 30 frames/s during the filling process. Image analysis is used to quantify the liquid distribution (center of mass, spatial standard deviation, skewness, kurtosis, and indicators of homogeneity) as time progresses. We compare the distribution dynamics in supine animals to those in upright animals for three constant infusion rates of perfluorocarbon: 15, 40, and 60 ml/min. It is found that formation of liquid plugs in large airways, which is affected by posture and infusion rate, can result in a more homogeneous liquid distribution than gravity drainage alone. The supine posture resulted in more homogeneous filling of the lungs than did upright posture, in which the lungs tend to fill in the inferior regions first. Faster instillation of perfluorocarbon results in liquid plugs forming in large airways and, consequently, more uniform distribution of perfluorocarbon than slower instillation rates in the upright animals.     

Beneficial effects of enhanced aeration using perfluorodecalin in <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> cultures for lipase production

The inadequate supply of oxygen to biomass is a critical factor to the productivity of most aerobic submerged fermentations. This happens because oxygen is sparingly soluble in the aqueous media. The use of a second liquid phase of perfluorocarbon (PFC), an oxygen-carrying compound, in the culture medium can increase the availability of oxygen to the microorganisms. The effect of perfluorodecalin on Yarrowia lipolytica cultures was investigated in shake-flask cultures. It was found that the specific growth rate of Y. lipolytica, a strictly aerobic yeast, increases with increasing PFC concentration. Extracellular lipase production was increased with 20% (v/v) of PFC and agitation of 250 rev/min. It was shown that the PFC presence benefitted lipase production and not just its secretion to the extracellular medium.     

Effects of Perfluorocarbons on surfactant exocytosis and membrane properties in isolated alveolar type II cells

Background

Perfluorocarbons (PFC) are used to improve gas exchange in diseased lungs. PFC have been shown to affect various cell types. Thus, effects on alveolar type II (ATII) cells and surfactant metabolism can be expected, data, however, are controversial.

Objective

The study was performed to test two hypotheses: (I) the effects of PFC on surfactant exocytosis depend on their respective vapor pressures; (II) different pathways of surfactant exocytosis are affected differently by PFC.

Methods

Isolated ATII cells were exposed to two PFC with different vapor pressures and spontaneous surfactant exocytosis was measured. Furthermore, surfactant exocytosis was stimulated by either ATP, PMA or Ionomycin. The effects of PFC on cell morphology, cellular viability, endocytosis, membrane permeability and fluidity were determined.

Results

The spontaneous exocytosis was reduced by PFC, however, the ATP and PMA stimulated exocytosis was slightly increased by PFC with high vapor pressure. In contrast, Ionomycin-induced exocytosis was decreased by PFC with low vapor pressure. Cellular uptake of FM 1-43 - a marker of membrane integrity - was increased. However, membrane fluidity, endocytosis and viability were not affected by PFC incubation.

Conclusions

We conclude that PFC effects can be explained by modest, unspecific interactions with the plasma membrane rather than by specific interactions with intracellular targets.     

Analysis of perfluorochemical elimination from the respiratory system

Shaffer, Thomas H., Raymond Foust IIII, Marla R. Wolfson,and Thomas F. Miller, Jr. Analysis of perfluorochemicalelimination from the respiratory system. J. Appl.Physiol. 83(3): 1033-1040, 1997.We describe asimple apparatus for analysis of perfluorochemicals (PFC) in expiredgas and thus a means for determining PFC vapor and liquid eliminationfrom the respiratory system. The apparatus and data analysis are basedon thermal conduction and mass transfer principles of gases. In vitrostudies were conducted with the PFC vapor analyzer to determinecalibration curves for output voltage as a function of individualrespiratory gases, respiratory gases saturated with PFC vapor, andvolume percent standards for percent PFC saturation (%PFC-Sat) in air.Voltage-concentration data for %PFC-Sat of the vapor from the in vitrotests were accurate to within 2.0% from 0 to 100% PFC-Sat, linear(r = 0.99, P < 0.001), and highly reproducible.Calculated volume loss of PFC liquid over time correlated well withactual loss by weight (r = 0.99, P < 0.001). In vivo studies withneonatal lambs demonstrated that PFC volume loss and evaporation ratesdecreased nonlinearly as a function of time. These relationships weremodulated by changes in PFC physical properties, minute ventilation,and postural repositioning. The results of this study demonstrate thesensitivity and accuracy of an on-line method for PFC analysis ofexpired gas and describe how it may be useful in liquid-assistedventilation procedures for determining PFC volume loss, evaporationrate, and optimum dosing and ventilation strategy.

     

Comparison of the pharmacokinetics,safety and tolerability of two concentrations of a new liquid recombinant human growth hormone formulation versus the freeze-dried formulation

Background  

Somatropin is recombinant human growth hormone (GH) used for the treatment of growth failure in children and GH deficiency in adults. Two concentrations of a liquid formulation have been developed: 5.83 and 8.0 mg/mL. This trial compared the pharmacokinetics (PK), safety and tolerability of these two liquid concentrations against the freeze-dried (FD) formulation in healthy volunteers.     

Application of perfluorocarbon emulsions for the administration of photosensitizing preparations into bone marrow stem cells

It has been shown that, upon incubation of mouse bone marrow stem cells (BMSC) in vitro with the nanoparticles of perfluorocarbon (PFC) emulsion stabilized by proxanol 268, these nanoparticles penetrate into cells and stay there for a long time (up to 20 days of observation). It has been found that, under in vitro conditions, mouse BMSC loaded with the nanoparticles of both the original emulsion and the emulsion preliminarily incubated with radachlorine do not differ from control stem cells in the rate of division, stretching on a plastic support, and the formation of a monolayer. It has been shown that the exposure to laser radiation of BMSC incubated with the nanoparticles of a PFC emulsion preliminarily incubated with radachlorine under in vitro conditions leads to the death of these cells due to the destruction of the cell membrane. The treatment with laser radiation of BMSC incubated with the nanoparticles of the starting PFC emulsion (without preliminarily incubation with radachlorine) causes no death of these cells. It has been shown in in vivo experiments that, when transplanted to the organism of a recipient mouse, BMSC of a donor mouse incubated with the nanoparticles of a PFC emulsion preliminarily incubated with radachlorine retain their functional activity, in particular the ability to migrate in the animal body. In this case, radachlorine contained in these stem cells retains its major function, to induce the death of stem cells by the action of laser radiation due to the destruction of the cell membrane. The observation period after the transplantation was 5–7 days.