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Attenuation of ocular wall pulsation in eyes containing a gas bubble after vitrectomy.

PURPOSE: To evaluate the effect of an intraocular gas bubble on ocular wall pulsation. METHODS: Eight patients that underwent vitrectomy with intraocular gas injection for various reasons were studied. Ocular pulsation was evaluated in a semi-quantitative way by means of Goldman applanation tonometry in both eyes before and after the operation. Vitreous cavity gas fill in the operated eye was estimated by means of indirect ophthalmoscopy and was correlated with ocular pulsation. The follow up lasted until the gas totally disappeared. RESULTS: Ocular pulsation disappeared postoperatively in the operated eye and gradually reappeared when the gas fill diminished. Pulsation returned to preoperative level when the gas disappeared from the vitreous cavity. A negative statistical correlation between ocular pulsation and gas fill could be documented. CONCLUSION: The compressible nature of the intraocular gas bubble results in absorption of the vascular pulsation and prevents its transfer to the ocular wall. This leads to attenuation or even cancellation of ocular wall pulsation when a gas bubble exists within the eye.

Eye↗

The degradability characteristics of fifty-four roughages and roughage neutral-detergent fibres as described by in vitro gas production and their relationship to voluntary feed intake.

Fifty-four roughages of known voluntary dry-matter intakes (DMI; range 7.8-35.2 g/kg live weight per d) were examined in vitro in a gas production test. Samples (200 mg) of roughage and roughage neutral-detergent fibre (NDF) respectively were incubated in a mixed suspension of rumen contents for 96 h and the gas volumes recorded after 4, 6, 8, 12, 24, 30, 36, 48, 54, 60 and 96 h. The kinetics of gas production were derived from the volume recordings described by the exponential equation Y = A + B(l-e-ct) where A is the intercept and ideally reflects the fermentation of the soluble and readily available fraction of the feed, B describes the fermentation of the insoluble (but with time fermentable) fraction and c the fractional rate at which B is fermented per h; A + B describes total fermentation. In vitro true dry matter (TD) and NDF degradabilities (NDF-D) after 24 h incubation were also determined. Of the variation in DMI, 75% was accounted for by the in vitro gas production parameters A, B and c in stepwise multiple regressions; 82% of the variation in DMI was explained by the parameters (ANDF + BNDF) and cNDF as obtained from the incubation of roughage NDF. The rate constants (c) were less important than parameters related to the extent of gas production, accounting for only 6.5 (whole roughage) and 4.1% (NDF) of the variation in DMI. There was no statistical advantage in the use of the exponential model describing extent and rate of fermentation over some of the simple gas volume measurements: 75% of the variation in DMI was accounted for by in vitro gas production of whole roughage after 8 h of incubation. On average gas production from NDF measured from 24-96 h accounted for 81% of the variation in DMI. A combination of gas volume measurements after a short period of incubation (4-8 h) with a concomitant determination of NDF-D after many hours (> or = 24 h) can render NDF preparations and long incubation times redundant. A method is suggested to obtain two results for DMI prediction in one single incubation. Of the variation in DMI 80% was accounted for by the incubation of 500 mg whole roughage when incubation was terminated after 24 h and the residual undegraded substrate quantified.

Animal Feed↗

Evaluation of soil gas sampling and analysis techniques at a former petrochemical plant site.

Methods for soil gas sampling and analysis are evaluated as part of a research study on soil vapour intrusion into buildings, conducted at a former petro-chemical plant site ("Chatterton site"). The evaluation process was designed to provide information on reliability and selection of appropriate methods for soil gas sampling and analysis, and was based on a literature review of data and methods, and experiments completed as part of the research study. The broader context of this work is that soil gas characterization is increasingly being used for input into risk assessment of contaminated sites, particularly when evaluating the potential intrusion of soil vapour into buildings. There are only a limited number of research studies and protocols addressing soil gas sampling and analysis. There is significant variability in soil gas probe design and sample collection and analysis methods used by practitioners. The experimental studies conducted to evaluate soil gas methods address the permeation or leakage of gases from Tedlar bags, time-dependent sorption of volatile organic compound (VOC)-vapours onto probe surfaces and sampling devices, and analytical and quality control issues for light gas and VOC analyses. Through this work, common techniques for soil gas collection and analysis are described together with implications for data quality arising from the different methods used. Some of the potential pitfalls that can affect soil gas testing are identified, and recommendations and guidance for improved protocols are provided.

Biodegradation, Environmental↗

A study for the thermal treatment of dehydrated sewage sludge with gas-agitated double screw type dryer.

A specially designed dryer has been developed for the thermal treatment of dehydrated, highly viscous sewage sludge with moisture content more than 80 wt.% by an gas-agitated, double-screw type dryer system. The treatment capacity of sludge dryer was about 100 kg/h. It consists of burner, feeding hopper, hot gas supplying ducts with double screw conveyor, damper and outlets of drying gas and dried material. The sewage sludge was transported by the revolution of the cylinder conveyor together with the tumbling and mixing action of the screw and lifters. The heating of the sludge was made efficient by the combination action of conduction and convection modes together with the gas-agitation process. The conduction occurred across the surface of the combustor cylinder, the convection was made by the flue gas interaction over the sludge and the agitation action through holes of conveyor cylinder into the bottom of the sludge. The number and location of gas-agitation holes and thereby the fraction of drying gas into the agitation holes were evaluated by numerical calculation of turbulent reacting flow. To evaluate the performance of the dryer developed in this study, a series of parametric experiments were performed in terms of important variables. In general drying process occurred successfully even for the highly agglomerating municipal sewage sludge through the first lumped plastic phase to a fine granulate state when the heat and mass balance was matched by adjusting rpm of the conveyor, burner capacity, and the fraction of gas-agitation. Further the avoidance of initial sludge agglomeration was considered to be important by the combination action of screw and lifter. The weight of the sludge was reduced approximately by 60% and therefore the volume of sludge was decreased to approximately 75% of initial volume with the water content of 10-20%. Energy efficiency of dryer was evaluated in the range of 70-75% at the sludge feed rate, 100 kg/h. The results obtained show the development of a robust, drying action applicable to highly viscous and high moisture content sewage sludge by the novel design of conveyor system and gas-agitation process developed in this study.

Algorithms↗

The secretion of inert gas into the swim-bladder of fish.

The composition of the gas mixture secreted into the swim-bladders of several species of fish has been determined in the mass spectrometer. The secreted gas differed greatly from the gas mixture breathed by the fish in the relative proportions of the chemically inert gases, argon, neon, helium, and nitrogen. Relative to nitrogen the proportion of the very soluble argon was increased and the proportions of the much less soluble neon and helium decreased. The composition of the secreted gas approaches the composition of the gas mixture dissolved in the tissue fluid. A theory of inert gas secretion is proposed. It is suggested that oxygen gas is actively secreted and evolved in the form of minute bubbles, that inert gases diffuse into these bubbles, and that the bubbles are passed into the swim-bladder carrying with them inert gases. Coupled to a preferential reabsorption of oxygen from the swim-bladder this mechanism can achieve high tensions of inert gas in the swim-bladder. The accumulation of nearly pure nitrogen in the swim-bladder of goldfish (Carassius auratus) is accomplished by the secretion of an oxygen-rich gas mixture followed by the reabsorption of oxygen.

Biological Transport↗

Independent control of blood gas PO2 and PCO2 in a bubble oxygenator.

An in-vitro circuit simulating cardiopulmonary bypass has been used to evaluate the gas transfer characteristics, gaseous microemboli production and blood damage of the Bentley-10 Plus bubble oxygenator. This oxygenator incorporates a unique integral gas controller which proportions the gas flow between the gas sparger and heat exchange area. The results of the study indicate that maintenance of a gas flow which ensures adequate CO2 removal can be prevented from inducing over-oxygenation by the manipulation of the gas controller which variably proportions the flow of gas from the oxygenation-efficient conventional gas sparger to the less efficient heat exchange area. Independent control of PaO2 with maintenance of CO2 transfer was demonstrated. Reduction in gaseous microemboli production and red blood cell damage with the use of the gas controller was demonstrated.

Blood Pressure↗

The carrier gas in anaesthesia: nitrous oxide/oxygen, medical air/oxygen and pure oxygen.

PURPOSE OF REVIEW: The gas passing the module for the delivery of inhalation anaesthetics and carrying vapourized anaesthetics into the breathing system is called the carrier gas. Oxygen is the absolutely indispensable component of the carrier gas. Additive gaseous components can be medical air (nitrogen), nitrous oxide, cyclopropane, or xenon, the latter three being anaesthetic gases themselves. Cyclopropane is not used any more and xenon is not approved as a medical gas yet, leaving medical air and nitrous oxide as the only currently available adjuncts to oxygen. RECENT FINDINGS: As review articles on nitrous oxide recommend unequivocally not using this gas routinely, and many anaesthetists already consistently omit any use of this gas, the alternative carrier gases are either a mixture of medical air and oxygen or pure oxygen. A nitrogen/oxygen mixture is absolutely inert with respect to metabolism or environment; there are no contraindications or shortcomings at all. The use of pure oxygen will result in high oxygen concentrations, improving the patients' safety by increasing the pulmonary oxygen reservoir and rendering impossible the development of hypoxic gas mixtures within the breathing system. Whether the incidence of postoperative wound infections or postoperative nausea and vomiting is reduced by perioperative hyperoxia is still under scientific discussion. In general, the use of high oxygen concentrations over a period of 6-8 h is not harmful for the patient, assuming the performance of recruitment manoeuvres, consistent use of positive end-expiratory pressure, and thorough observance of the contraindications that can result from the increased vulnerability of tissues to hyperoxia. SUMMARY: Nitrous oxide should not be used routinely as a component of the carrier gas any more. A mixture of medical air and oxygen must be acknowledged to be the gold standard. Pure oxygen may be used as a carrier gas if medical air or properly performing flow controls for medical air are not available.

Journal Article↗

Perfluorocarbon-associated gas exchange improves oxygenation, lung mechanics, and survival in a model of adult respiratory distress syndrome.

OBJECTIVE: To compare the effectiveness of perfluorocarbon-associated gas exchange to volume controlled positive pressure breathing in supporting gas exchange, lung mechanics, and survival in an acute lung injury model. DESIGN: A prospective, randomized study. SETTING: A university medical school laboratory approved for animal research. SUBJECTS: Neonatal piglets. INTERVENTIONS: Eighteen piglets were randomized to receive perfluorcarbon-associated gas exchange with perflubron (n=10) or volume controlled continuous positive pressure breathing (n=8) after acute lung injury was induced by oleic acid infusion (0.15 mL/kg iv). MEASUREMENTS AND MAIN RESULTS: Arterial and venous blood gases, hemodynamics, and lung mechanics were measured every 15 mins during a 3-hr study period. All animals developed a metabolic and a respiratory acidosis during the infusion of oleic acid. Following randomization, the volume controlled positive pressure breathing group developed a profound acidosis (p<.05), while pH did not change in the perfluorocarbon-associated gas exchange group. Within 15 mins of initiating perfluorocarbon-associated gas exchange, oxygenation increased from a PaO2 of 52 +/- 12 torr (6.92 +/- 1.60 kPa) to 151 +/- 93 torr (20.0 +/- 12.4 kPa) and continued to improve throughout the study (p<.05). Animals that received volume controlled positive pressure breathing remained hypoxic with no appreciable change in PaO2. Although both groups developed hypercarbia during oleic acid infusion, PaCO2, steadily increased over time in the control group (p<.01). Static lung compliance significantly increased postrandomization (60 mins) in the animals supported by perflurocarbon-associated gas exchange (p<.05), whereas it remained unchanged over time in the volume controlled positive pressure breathing group. However, survival was significantly higher in the perfluorocarbon-associated gas exchange group with eight (80%) of ten animals surviving the entire study period. Only two (25%) of the eight animals in the volume controlled positive pressure breathing group were alive at the end of the study period (log-rank statistic, p=.013). CONCLUSIONS: Perflurocarbon-associated gas exchange enhanced gas exchange, pulmonary mechanics, and survival in this model of acute lung injury.

Animals↗

Helium versus oxygen for tracheal gas insufflation during mechanical ventilation.

OBJECTIVE: To evaluate and compare the effect of tracheal gas insufflation using two gases with different physical properties, helium and oxygen, as an adjunct to conventional mechanical ventilation in patients with respiratory failure. DESIGN: Prospective, intervention study. SETTING: General intensive care unit in a tertiary university medical center. PATIENTS: Seven sedated and paralyzed patients with respiratory failure of various etiologies. All patients were ventilated in the volume-control mode (tidal volume 5 to 7 mL/kg). Inclusion criteria were PaCO2 of > or =50 torr (> or =6.7 kPa), together with peak inspiratory pressure of > or =35 cm H2O and respiratory rate of > or =14 breaths/min. INTERVENTIONS: All patients were intubated with an endotracheal tube that had an additional lumen opening at its distal end, through which tracheal gas insufflation was administered. The tracheal gas insufflation was applied continuously throughout the respiratory cycle at three flow rates (2, 4, and 6 L/min) with two gases, oxygen and helium, while the ventilatory settings were maintained constant. MEASUREMENTS AND MAIN RESULTS: In addition to airway pressures and arterial blood gases, the relative efficacy of tracheal gas insufflation with each gas was estimated using a "coefficient of efficiency" (which we defined as the change in PaCO2/peak inspiratory pressure) compared with baseline measurements. Tracheal gas insufflation with both gases decreased PaCO2 significantly (p < .05) at all flow rates. This effect was accompanied by an increase in airway pressure with both gases (oxygen and helium). However, at flow rates of 6 L/min, tracheal gas insufflation with helium resulted in lower peak inspiratory pressure than with oxygen. Tracheal gas insufflation with helium was more effective (as estimated by the coefficient of efficiency) than with oxygen at all flow rates (p < .05). CONCLUSION: In volume-controlled, mechanically ventilated patients with respiratory failure, tracheal gas insufflation with helium might be suggested as an alternative to oxygen.

Acute Disease↗

A fast, digitally controlled flow proportional gas injection system for studies in lung function.

The aim of this paper is to describe a device for flow proportional injection of tracer gas in the lungs of mechanically ventilated patients. This device may then be used for the study of the multiple breath indicator gas washout technique to determine the end-expiratory lung volume. Such a tracer gas injection device may also be used in the study of other techniques that rely on uptake and elimination of tracer gas by the lungs. In this paper, an injector is described which enables injection of indicator gas at a predetermined concentration in a breathing circuit independent of the type of breathing. The presented setup uses a control computer to produce steering signals to a multivalve array in proportion to the input breathing signals. The multivalve array consists of ten circular valves, each with a different diameter, which can be opened or closed individually according to the input signal of the array. By opening of a certain combination of valves an amount of sulphur hexafluoride gas proportional to the inspiratory breathing signal is released. The rate of transmission between the components of the injection system was 80 Hz. The injector has a full flow range between 0-10 L/min. The delay time between the breathing signal and the flow response was 70 ms. The aimed washin gas concentration of 1% SF6 was achieved after 0.5 s. The study describes the results of tests to determine valve-flow ratios, step response and dynamic response of the injector. The flow output response of the injector system was shown to increase in input frequencies above 3 Hz. The valve flow ratios showed the largest relative deviation in the two smallest valves of the 10 valve array, respectively 0.005 L/min (25%) and 0.002 L/min (20%). We conclude that the injector can achieve a stable concentration of indicator gas in a breathing system with an accuracy of 0.005 L/min to execute the multiple breath indicator washout test in human subjects. The results of the study indicate that the injector may be of use in other application fields in respiratory physiology in which breathing circuit injection of indicator gas is required.

Algorithms↗

Intestinal tone and gas motion.

The intestine propels and evacuates large gas loads without detectable phasic contractions by manometry. We hypothesized that intestinal gas motion is produced by changes in gut tone and capacitance. In 13 healthy subjects, changes in duodenal tone were measured by a barostat during continuous perfusion of lipids (Intralipid, 1 kcal min(-1)) into the duodenum for 60 min. In separate groups, the effects of jejunal gas infusion (N2, CO2 and O2 in venous proportions at 12 mL min(-1) starting after 15 min lipid perfusion) and sham infusion were tested. Gas outflow was collected continuously via an intrarectal cannula. Duodenal lipid perfusion produced a rapid duodenal relaxation (volume increased by 48 +/- 18%; P < 0.01 vs basal). Gas infusion increased gas evacuation (184 +/- 59 mL), and this was associated with a tonic contraction of the duodenum (R = 0.86; P < 0.01) that completely reverted the lipid-induced duodenal relaxation (volume decreased by 42 +/- 13%; P < 0.05). During sham infusion only 52 +/- 28 mL of gas were evacuated (P < 0.05 vs gas infusion), and the duodenum remained relaxed due to the effect of lipids (0 +/- 1% volume reduction; ns). In conclusion, intestinal gas propulsion and clearance is associated with a tonic contraction of the gut wall and reduced gut capacitance.

Adult↗

Applications of a total dissolved gas pressure probe in ground water studies.

Measurements of dissolved gases have numerous applications in ground water hydrology, and it is now possible to measure the total dissolved gas pressure in situ using a probe. Dissolved gas pressure is measured by submerging a headspace volume with a gas-permeable membrane, allowing dissolved gases in the water to equilibrate with gases in the headspace, then measuring the pressure in the headspace with a pressure transducer. Total dissolved gas pressure (TGP) probes have many potential uses in ground water studies employing dissolved gases, including: (1) determining approximate excess air levels, which may provide information about the time and location of recharge; (2) screening wells for air contamination, which can compromise the accuracy of dissolved gas tracer techniques: (3) detecting a trapped gas phase, which can significantly reduce hydraulic conductivity and impede the transport of dissolved solutes and gases; (4) enabling the use of gas-filled passive diffusion samplers for determining accurate dissolved gas concentrations; and (5) determining relative concentrations of CH4 and CO2 when they are known to be highly abundant. Although TGP probes designed for surface water have been available for several years, TGP probes suitable for ground water applications have only recently become available. Herein we present what are, to our knowledge, the first reported ground water dissolved gas data collected using a TGP probe. We also explain the basic operating principles of these probes and discuss the potential applications listed.

Carbon Dioxide↗

Intrapulmonary receptor response to changes in airway-gas composition in Gallus domesticus.

1. A unidirectional, artificial ventilating system was used to observe respiratory and neural responses to changes in CO(2) concentration in pulmonary airways of the chicken.2. Sectioning any of the three pulmonary branches of the vagus slowed the respiratory response to sudden removal of CO(2) from unidirectional ventilating gas stream.3. The impulse frequency of CO(2)-sensitive units was inversely related to the CO(2) content in the ventilating gas stream.4. Abrupt elimination of CO(2) from the ventilatory gas stream caused a transient burst of activity from CO(2)-sensitive receptors, which quickly adapted to a constant discharge frequency. Fifty percent of the units responded within 0.35 sec after CO(2)-free gas reached the lungs. The neural response to abrupt re-addition of CO(2) to the gas stream was less rapid.5. Acetylcholine or NaCN injected into the pulmonary artery had no apparent effect on the discharge of CO(2)-sensitive receptors. Veratridine temporarily reduced discharge and occasionally produced short, rapid bursts of activity.6. Hypoxia and hyperoxia (5% and 80% O(2)) produced no significant change in the discharge of CO(2)-sensitive receptors.7. Carbon dioxide-sensitive units in spontaneously breathing chickens were silenced by adding 10% CO(2) to the inspired gas; the units responded vigorously to inflation of the respiratory system with a gas containing no CO(2) but remained silent during inflation with gas containing 15% CO(2). Units insensitive to CO(2), which increased their impulse frequency during inflation regardless of the CO(2) content of the inflating gas, also were observed.

Acetylcholine↗

Mathematical model of gas bubble evolution in a straight tube.

Deep sea divers suffer from decompression sickness (DCS) when their rate of ascent to the surface is too rapid. When the ambient pressure drops, inert gas bubbles may form in blood vessels and tissues. The evolution of a gas bubble in a rigid tube filled with slowly moving fluid, intended to simulate a bubble in a blood vessel, is studied by solving a coupled system of fluid-flow and gas transport equations. The governing equations for the fluid motion are solved using two techniques: an analytical method appropriate for small nondeformable spherical bubbles, and the boundary element method for deformable bubbles of arbitrary size, given an applied steady flow rate. A steady convection-diffusion equation is then solved numerically to determine the concentration of gas. The bubble volume, or equivalently the gas mass inside the bubble for a constant bubble pressure, is adjusted over time according to the mass flux at the bubble surface. Using a quasi-steady approximation, the evolution of a gas bubble in a tube is obtained. Results show that convection increases the gas pressure gradient at the bubble surface, hence increasing the rate of bubble evolution. Comparing with the result for a single gas bubble in an infinite tissue, the rate of evolution in a tube is approximately twice as fast. Surface tension is also shown to have a significant effect. These findings may have important implications for our understanding of the mechanisms of inert gas bubbles in the circulation underlying decompression sickness.

Atmospheric Pressure↗

Measurement of biologically available naphthalene in gas and aqueous phases by use of a Pseudomonas putida biosensor.

Genetically constructed microbial biosensors for measuring organic pollutants are mostly applied in aqueous samples. Unfortunately, the detection limit of most biosensors is insufficient to detect pollutants at low but environmentally relevant concentrations. However, organic pollutants with low levels of water solubility often have significant gas-water partitioning coefficients, which in principle makes it possible to measure such compounds in the gas rather than the aqueous phase. Here we describe the first use of a microbial biosensor for measuring organic pollutants directly in the gas phase. For this purpose, we reconstructed a bioluminescent Pseudomonas putida naphthalene biosensor strain to carry the NAH7 plasmid and a chromosomally inserted gene fusion between the sal promoter and the luxAB genes. Specific calibration studies were performed with suspended and filter-immobilized biosensor cells, in aqueous solution and in the gas phase. Gas phase measurements with filter-immobilized biosensor cells in closed flasks, with a naphthalene-contaminated aqueous phase, showed that the biosensor cells can measure naphthalene effectively. The biosensor cells on the filter responded with increasing light output proportional to the naphthalene concentration added to the water phase, even though only a small proportion of the naphthalene was present in the gas phase. In fact, the biosensor cells could concentrate a larger proportion of naphthalene through the gas phase than in the aqueous suspension, probably due to faster transport of naphthalene to the cells in the gas phase. This led to a 10-fold lower detectable aqueous naphthalene concentration (50 nM instead of 0.5 micro M). Thus, the use of bacterial biosensors for measuring organic pollutants in the gas phase is a valid method for increasing the sensitivity of these valuable biological devices.

Bacterial Proteins↗

Comparative study of the structure of gas vacuoles.

The fine structure of gas vacuoles was examined in two blue-green algae, two green bacteria, three purple sulfur bacteria, and two halobacteria. The gas vacuole is a compound organelle, composed of a variable number of gas vesicles. These are closed, cylindrical, gas-containing structures with conical ends, about 80 to 100 nm in width and of variable length, ranging from 0.2 to over 1.0 mum. The wall of the gas vesicle is a non-unit membrane 2 to 3 nm in thickness, bearing very regular striations with a periodicity of 4 nm, oriented more or less at right angles to the long axis of the cylinder. This fine structure could be clearly resolved in isolated gas vesicles prepared from a blue-green alga and from Halobacterium halobium, and its presence in the gas vesicles of the green bacterium Pelodictyon clathratiforme was inferred from thin sections. The gas vacuole thus appears to be a homologous organelle in all of these procaryotic groups. Minor differences with respect to the length and arrangement of the gas vesicles were observed. In blue-green algae and green bacteria, the vesicles are relatively long and tend to be arrayed in parallel bundles; in purple sulfur bacteria and Halobacterium, they are shorter and more irregularly distributed in the cell.

Bacteria↗

Eight of fourteen gvp genes are sufficient for formation of gas vesicles in halophilic archaea.

The minimal number of genes required for the formation of gas vesicles in halophilic archaea has been determined. Single genes of the 14 gvp genes present in the p-vac region on plasmid pHH1 of Halobacterium salinarum (p-gvpACNO and p-gvpDEFGHIJKLM) were deleted, and the remaining genes were tested for the formation of gas vesicles in Haloferax volcanii transformants. The deletion of six gvp genes (p-gvpCN, p-gvpDE, and p-gvpHI) still enabled the production of gas vesicles in H. volcanii. The gas vesicles formed in some of these gvp gene deletion transformants were altered in shape (Delta I, Delta C) or strength (Delta H) but still functioned as flotation devices. A minimal p-vac region (minvac) containing the eight remaining genes (gvpFGJKLM-gvpAO) was constructed and tested for gas vesicle formation in H. volcanii. The minvac transformants did not form gas vesicles; however, minvac/gvpJKLM double transformants contained gas vesicles seen as light refractile bodies by phase-contrast microscopy. Transcript analyses demonstrated that minvac transformants synthesized regular amounts of gvpA mRNA, but the transcripts derived from gvpFGJKLM were mainly short and encompassed only gvpFG(J), suggesting that the gvpJKLM genes were not sufficiently expressed. Since gvpAO and gvpFGJKLM are the only gvp genes present in minvac/JKLM transformants containing gas vesicles, these gvp genes represent the minimal set required for gas vesicle formation in halophilic archaea. Homologs of six of these gvp genes are found in Anabaena flos-aquae, and homologs of all eight minimal halobacterial gvp genes are present in Bacillus megaterium and in the genome of Streptomyces coelicolor.

Archaeal Proteins↗

Complexity of gas vesicle biogenesis in Halobacterium sp. strain NRC-1: identification of five new proteins.

The genome of Halobacterium sp. strain NRC-1 contains a large gene cluster, gvpMLKJIHGFEDACNO, that is both necessary and sufficient for the production of buoyant gas-filled vesicles. Due to the resistance of gas vesicles to solubilization, only the major gas vesicle protein GvpA and a single minor protein, GvpC, were previously detected. Here, we used immunoblotting analysis to probe for the presence of gas vesicle proteins corresponding to five additional gvp gene products. Polyclonal antisera were raised in rabbits against LacZ-GvpF, -GvpJ, and -GvpM fusion proteins and against synthetic 15-amino-acid peptides from GvpG and -L. Immunoblotting analysis was performed on cell lysates of wild-type Halobacterium sp. strain NRC-1, gas vesicle-deficient mutants, and purified gas vesicles, after purification of LacZ fusion antibodies on protein A and beta-galactosidase affinity columns. Our results show the presence of five new gas vesicle proteins (GvpF, GvpG, GvpJ, GvpL, and GvpM), bringing the total number of proteins identified in the organelles to seven. Two of the new gas vesicle proteins are similar to GvpA (GvpJ and GvpM), and two proteins contain predicted coiled-coil domains (GvpF and GvpL). GvpL exhibited a multiplet ladder on sodium dodecyl sulfate-polyacrylamide gels indicative of oligomerization and self-assembly. We discuss the possible functions of the newly discovered gas vesicle proteins in biogenesis of these unique prokaryotic flotation organelles.

Amino Acid Sequence↗