Microbial processes in the Athabasca Oil Sands and their potential applications in microbial enhanced oil recovery.

The Athabasca Oil Sands are located within the Western Canadian Sedimentary Basin, which covers over 140,200 km(2) of land in Alberta, Canada. The oil sands provide a unique environment for bacteria as a result of the stressors of low water availability and high hydrocarbon concentrations. Understanding the mechanisms bacteria use to tolerate these stresses may aid in our understanding of how hydrocarbon degradation has occurred over geological time, and how these processes and related tolerance mechanisms may be used in biotechnology applications such as microbial enhanced oil recovery (MEOR). The majority of research has focused on microbiology processes in oil reservoirs and oilfields; as such there is a paucity of information specific to oil sands. By studying microbial processes in oil sands there is the potential to use microbes in MEOR applications. This article reviews the microbiology of the Athabasca Oil Sands and the mechanisms bacteria use to tolerate low water and high hydrocarbon availability in oil reservoirs and oilfields, and potential applications in MEOR.

The role of nutrient loading and eutrophication in estuarine ecology.

Eutrophication is a process that can be defined as an increase in the rate of supply of organic matter (OM) to an ecosystem. We provide a general overview of the major features driving estuarine eutrophication and outline some of the consequences of that process. The main chemical constituent of OM is carbon (C), and therefore rates of eutrophication are expressed in units of C per area per unit time. OM occurs in both particulate and dissolved forms. Allochthonous OM originates outside the estuary, whereas autochthonous OM is generated within the system, mostly by primary producers or by benthic regeneration of OM. The supply rates of limiting nutrients regulate phytoplankton productivity that contributes to inputs of autochthonous OM. The trophic status of an estuary is often based on eutrophication rates and can be categorized as oligotrophic (<100 g C m(-2) y(-1), mesotrophic (100-300 g C m(-2) y(-1), eutrophic (300-500 g C m(-2) y(-1), or hypertrophic (>500 g C m(-2) y(-1). Ecosystem responses to eutrophication depend on both export rates (flushing, microbially mediated losses through respiration, and denitrification) and recycling/regeneration rates within the estuary. The mitigation of the effects of eutrophication involves the regulation of inorganic nutrient (primarily N and P) inputs into receiving waters. Appropriately scaled and parameterized nutrient and hydrologic controls are the only realistic options for controlling phytoplankton blooms, algal toxicity, and other symptoms of eutrophication in estuarine ecosystems.

Ecosystem impacts of three sequential hurricanes (Dennis, Floyd, and Irene) on the United States' largest lagoonal estuary, Pamlico Sound, NC.

Three sequential hurricanes, Dennis, Floyd, and Irene, affected coastal North Carolina in September and October 1999. These hurricanes inundated the region with up to 1 m of rainfall, causing 50- to 500-year flooding in the watershed of the Pamlico Sound, the largest lagoonal estuary in the United States and a key West Atlantic fisheries nursery. We investigated the ecosystem-level impacts on and responses of the Sound to the floodwater discharge. Floodwaters displaced three-fourths of the volume of the Sound, depressed salinity by a similar amount, and delivered at least half of the typical annual nitrogen load to this nitrogen-sensitive ecosystem. Organic carbon concentrations in floodwaters entering Pamlico Sound via a major tributary (the Neuse River Estuary) were at least 2-fold higher than concentrations under prefloodwater conditions. A cascading set of physical, chemical, and ecological impacts followed, including strong vertical stratification, bottom water hypoxia, a sustained increase in algal biomass, displacement of many marine organisms, and a rise in fish disease. Because of the Sound's long residence time ( approximately 1 year), we hypothesize that the effects of the short-term nutrient enrichment could prove to be multiannual. A predicted increase in the frequency of hurricane activity over the next few decades may cause longer-term biogeochemical and trophic changes in this and other estuarine and coastal habitats.

Minimum time interval adjustment for 4-3-1 immunization rates among two-year-old children.

BACKGROUND: The purpose is to determine the administrative validity of the 4-3-1 immunization rates for DPT-OPV-MMR vaccines for 2-year-old children in a community health status assessment project by accounting for premature administration of specific vaccine doses according to ACIP recommended minimal timing intervals. METHODS: A retrospective survey of immunization certificates was made on a random sample of 1,059 kindergarten enrollees in the county, including public, private, and parochial schools. The immunization rates by the crude 4-3-1 counting method were compared with the same method adjusted for minimal time interval vaccine dosing. RESULTS: By the crude 4-3-1 counting method, 55.5% of the students had complete immunizations by their second birthdate, and 11.8% did not meet the minimum interval recommendations on at least one of 4-3-1 vaccine series. The adjustment for minimum time interval reduced the percent in compliance with the 4-3-1 counting method by age 2 in the community to 50.7%. CONCLUSION: The premature timing of vaccine doses is a threat to the validity of the 4-3-1 counting method. The crude 4-3-1 method over-estimates the completed immunization rates for 2-year-olds in this community-based study by about 4.8%.

DXA body composition: theory and practice.

Dual energy X-ray absorptiometry (DXA) was used to measure human body composition on a high speed fan beam scanner. High and low energy attenuation pairs, produced by the various combinations of fat mass and fat-free mass in the human body, were compared to attenuation values produced by standard materials (aluminum and acrylic). These standards were measured in various combinations to construct calibration curves for fat and fat-free mass. Primary calibration of the aluminum/acrylic combinations was achieved by direct comparison to the dual energy attenuation produced by stearic acid and pure water. Whole body examinations were accomplished using three 45 s longitudinal passes of the fan beam. These passes were acquired and assembled to create a giant, isocentric fan beam with a single center of focus. In vivo precision was 0.009 g/cm2 for BMD and 425 g for fat mass and fat-free mass (s.d.).

Postsynaptic action potentials do not alter short-term potentiation in the dentate gyrus.

The dendrites of neocortical neurons have been shown to support active action potentials which back-propagate from the soma after an output spike has been initiated. This observation has led to speculation that dendritic action potentials may participate in various forms of synaptic plasticity. The contribution of dendritic spikes to paired-pulse facilitation (PPF), a form of short-term plasticity, was investigated in the dentate gyrus of hippocampal slices. Paired orthodromic stimulation of the perforant path produced an average facilitation of the test population spike (PS) amplitude of 167% (n = 16, conditioning response = 100%). There was also a small but significant increase in slope of the field EPSP (fEPSP) of 108%. To determine whether increased presynaptic drive could account for this facilitation, the relationship between fEPSP slope and spike amplitude (I-O) was determined for a range of stimulus intensities. An increase in fEPSP slope of 171% was associated with an increase in PS amplitude equal to the facilitation produced by paired-pulse stimulation (167%), suggesting a postsynaptic component in PPF. Electric field effects were then used as a tool to alter the excitability of granule cells during the conditioning response without changing synaptic drive. Any change in the test response associated with manipulation of the conditioning population spike amplitude would suggest that dendritic spikes may contribute to the postsynaptic component of PPF. Surprisingly, altering the number of neurons responding to the conditioning stimulus with an action potential had no effect on the test response, suggesting that dendritic action potentials do not participate in this form of short-term synaptic plasticity.

African-American smokers and cancers of the lung and of the upper respiratory and digestive tracts. Is menthol part of the puzzle?

The prevalence of cigarette smoking is higher among African Americans than among whites. African Americans have higher rates of lung cancer than whites, although they smoke fewer cigarettes. To explore this black-white difference in lung cancer rates, I examine various aspects of tobacco use in African-American smokers, including the age of initiation of smoking, quantity of cigarettes smoked, quit rates, level of nicotine dependence, biochemical differences, and brand preferences, specifically menthol brand cigarettes. I also review briefly the sequelae of patterns of tobacco use, including rates of lung and other tobacco-related cancers. A preference for mentholated cigarettes by African Americans is well documented and is one of the most striking differences between African-American and white smokers. Menthol brand preference has been investigated in an attempt to explain the black-white differences in rates of cancers of the lungs and the upper respiratory and digestive tracts. Also, studies have evaluated smoking behavior both with and without menthol and have explicitly examined the question of whether menthol use helps explain the black-white difference in lung cancer rates. The results of these studies are so far inconclusive with regard to the use of menthol and the risk of lung cancer developing. I provide practical suggestions for clinicians in counseling African-American smokers to quit smoking and to maintain a nonsmoking status.

A practical vibration isolation workstation for electrophysiology.

Vibration control is a major concern in electrophysiological research, particularly during intracellular recording where movements of only a few micrometers may disrupt the cell membrane. The workstation described here is based on the concept of a table nested within a second table. The inner table supports a vibration-isolated surface while the outer table provides protection and a bench-top for equipment. The work surface is supported on squash balls in order to avoid the substantial cost of passive commercial isolators. The properties of the squash ball isolators were found to be similar to a set commercial isolators when compared with respect to impulse response, compliance and transmissibility. The table has been successfully used for both intra- and extracellular recording over a 3-year period. Work efficiency was enhanced by the close proximity of the bench-top and isolated work surface, while perimeter protection helped maintain stable recordings.

Epileptiform activity in the dentate gyrus during low-calcium perfusion and exposure to transient electric fields.

1. The dentate gyrus fails to develop epileptiform activity in many experimental models of epilepsy, including the in vitro low-Ca2+ model. Although manipulating the K+ concentration or osmolality of normal low-Ca2+ perfusion mediums can enhance the propensity of the dentate gyrus to develop seizure activity, the specific mechanisms contributing to this change are still under investigation. Identification of these mechanisms should improve our understanding of epileptogenesis and of the factors contributing to the propensity for seizure discharge in other tissues. 2. In the present experiments we used externally generated electric fields to depolarize the somata of large populations of dentate granule cells during exposure to a perfusion medium with no added Ca2+ (low-Ca2+ medium). Uniform electric fields were generated across an individual slice by passing current between two parallel AgCl-coated silver wires placed on the surface of the artificial cerebral spinal fluid. The wires were positioned to straddle the slice such that the current flow was parallel to the dendrosomatic axis of the cell population under investigation. 3. Under control conditions (low-Ca2+ medium, no applied field), stimulation of the dentate hilus evoked a single antidromic population spike in 89% of the slices studied (n = 27). During application of electric fields the same stimulus evoked epileptiform activity in all trials. Well-formed bursts first occurred at an average field intensity of +22.9 +/- 2.5 (SE) mV/mm (n = 24). The amplitude of individual spikes and the total number of spikes, within a burst increased in a graded fashion as the magnitude of the applied field was increased. 4. High field intensities evoked epileptiform activity in the absence of a synchronizing antidromic stimulus. These field-induced bursts occurred after a progressive buildup of rhythmic activity recorded in the extrasomatic space and could persist for the entire duration of an applied field, lasting for several seconds. The average field intensity required to produce a threshold burst was +84.6 +/- 3.6 mV/mm (n = 24). 5. In 11% of trials (3 of 27) the dentate gyrus exhibited poorly developed antidromic bursting without the application of depolarizing electric fields. These bursts were completely suppressed by hyperpolarizing fields in the range of -10 to -20 mV/mm. 6. The results of this investigation support the hypothesis that granule cell sensitivity to nonsynaptic interactions is adequate to support bursting in a normal low-Ca2+ medium, but bursting fails to occur because these cells are normally too hyperpolarized relative to their action potential threshold.(ABSTRACT TRUNCATED AT 400 WORDS)

Thy1 (+) and (-) lung fibrosis subpopulations in LEW and F344 rats.

Appreciation of the potential of fibroblasts as effector cells in inflammation has led to the recognition of fibroblast subpopulations, the most stable of which are the Thy1 (+) and Thy1 (-) subpopulations in mouse lung fibroblasts. We investigated the presence of Thy1 (+) and (-) fibroblasts in rats, comparing the percentage in primary cultures from rats with different susceptibility to fibrosis, and whether the characteristics were similar in mice and rats, and between normal and fibrotic rats lungs. Using primary cultures of rat fibroblasts obtained both from normal and fibrotic lungs, we analysed the percentage of Thy1 (+) and (-) fibroblasts by fluorescence-activated cell sorter (FACS) analysis. We sorted the fibroblasts to evaluate immune region associated antigen (Ia) expression, which tends to be raised in tissues involved in inflammation, and other characteristics. We found that Thy1 (+) and (-) fibroblasts: 1) are distinct subpopulations in rat lungs; 2) are found in different proportions in rat strains with different propensity towards lung fibrosis; and 3) have similar but not identical characteristics in mice and rats. We also found that bleomycin-induced fibrosis increases the percentage of Ia expression in Thy1 (-), but not Thy1 (+) fibroblasts. The presence of these stable fibroblast supopulations in multiple species, and the fact that these fibroblasts differ in their response to a fibrosing agent, suggests the importance of considering fibroblast subpopulations in development and disease.

The site for initiation of action potential discharge over the somatodendritic axis of rat hippocampal CA1 pyramidal neurons.

Early electrophysiological studies in the mammalian hippocampus reported that orthodromic depolarization of pyramidal cells evoked action potential discharge (presumed Na+ dependent) both at the axon hillock and at one or more sites in the dendritic arborization (Cragg and Hamlyn, 1955; Andersen, 1959, 1960; Spencer and Kandel, 1961; Andersen and Lomo, 1966). Although tetrodotoxin (TTX)-sensitive spikes have been recorded at the dendritic level (Wong et al., 1979; Benardo et al., 1982; Miyakawa and Kato, 1986; Turner et al., 1989), the site for initiation of these potentials has not yet been determined. In this study, we examine the site for initiation of Na+ spike discharge over the cell axis of rat hippocampal CA1 pyramidal neurons. Intrasomatic and intradendritic recordings were obtained from pyramidal neurons of hippocampal slices maintained in vitro. Spike discharge was evoked by alvear (antidromic) stimulation or orthodromically by stimulation of afferent inputs in stratum oriens (SO) or stratum radiatum (SR). Antidromic and orthodromic spikes were greatest in amplitude in somatic recordings and declined over the apical dendritic axis, while spike half-width was shortest at the cell body and increased with distance from stratum pyramidale. Measurements of orthodromic spike threshold revealed that the only location at which spikes discharged at a consistent membrane potential at threshold intensity (voltage threshold) was the cell body region. Finally, at threshold intensity, SR-evoked intradendritic spikes were blocked by local application of TTX in stratum pyramidale, while spike blockade at suprathreshold intensity required the diffusion of TTX into the apical dendritic region. These results indicate that, for threshold intensities of stimulation, antidromic and orthodromic spike discharge in CA1 pyramidal cells is initiated in the region of the cell body layer, subsequently conducting over the apical dendrites in a retrograde fashion. In contrast, SR-evoked orthodromic spike discharge exhibits an intensity-dependent shift in the site of origin up to 200 microns within the apical dendritic arborization.

Apical dendritic depolarizations and field interactions evoked by stimulation of afferent inputs to rat hippocampal CA1 pyramidal cells.

The relationship between orthodromic extracellular field potentials and intradendritic depolarizations in apical dendrites of CA1 pyramidal neurons was investigated using the in vitro slice preparation of rat hippocampus. Orthodromic synaptic field potentials evoked by stimulation of afferent inputs in stratum radiatum or stratum oriens were used to measure extracellular voltage gradients generated over the pyramidal cell axis. Extracellular gradients were of opposite polarity over the region of pyramidal cell apical dendrites in stratum radiatum. The stratum radiatum-evoked gradient was negative towards the apical dendrites and the stratum oriens-evoked gradient negative towards the cell body layer, with gradients reaching values of up to 50 mV/mm over the apical dendritic axis. Intradendritic recordings obtained greater than 150 microns from stratum pyramidale directly measured the subthreshold apical dendritic excitatory postsynaptic potentials evoked by stratum radiatum or stratum oriens stimulation. These ground-referenced recordings were then compared to the transmembrane potential calculated by subtraction of the corresponding extradendritic field potential. Both stratum radiatum and stratum oriens stimulation evoked graded excitatory postsynaptic potentials that could be recorded in apical dendritic impalements up to 265 microns from stratum pyramidale. The calculated transmembrane potential of the stratum radiatum-evoked excitatory postsynaptic potential had a significantly greater rate of rise, peak amplitude, and rate of decay than that of the ground-referenced excitatory postsynaptic potential. In contrast, the rates of rise and decay of the transmembrane potential of the stratum oriens-evoked excitatory postsynaptic potential were reduced with respect to the ground-referenced recording. The peak amplitude of the stratum oriens-evoked transmembrane potential, however, varied according to the polarity of the corresponding extradendritic population spike response recorded in stratum radiatum. These data reveal that synaptic activation of either basal or apical dendrites of CA1 pyramidal cells evokes a depolarization that can be recorded over a substantial region of the apical dendritic arbor. Furthermore, extradendritic field potentials evoked by stimulation of these inputs produce opposite effects on the transmembrane potential of apical dendrites. The magnitude of the accompanying extracellular voltage gradients suggest that these shifts in transmembrane potential reflect ephaptic interactions at the apical dendritic level of pyramidal cells.

Heligmosomoides polygyrus: effect of exogenous steroid hormones on egg output in vitro.

The effect of exogenous steroid hormones on the egg output of Heligmosomoides polygyrus (Nematoda: Trichostrongylidae) was examined in vitro. Using worms raised in female mice, it was found that estradiol, testosterone, and cortisone each significantly decreased egg output. Although similar trends were found using H. polygyrus raised in male mice, none of the decreases found was significant. No significant differences were found with ecdysone or progesterone treatments using worms from female or male mice. Treatment of worms with cortisone did not significantly affect retention of eggs within the uterus of H. polygyrus. Titration of the effect of cortisone on egg output indicated that levels of reduction were significant for concentrations of 5.6 x 10(-6) M to 5.6 x 10(-3) M in worms from female mice and for concentrations of 5.6 x 10(-8), x 10(-7), x 10(-5) and x 10(-3) in worms from male mice. Radioisotope labelling experiments showed incorporation of 3H-corticosterone in the nucleus of intestinal cells of H. polygyrus suggesting that its effect on egg production may be via a modulatory effect on the intestinal cells.

Action-potential discharge in hippocampal CA1 pyramidal neurons: current source-density analysis.

1. The site of origin of evoked action-potential discharge in hippocampal CA1 pyramidal neurons was investigated using the in vitro rat hippocampal slice preparation. 2. Action-potential discharge in pyramidal cells was evoked by stimulation of efferent pyramidal cell fibers in the alveus (antidromic) or afferent synaptic inputs in stratum oriens (SO) or stratum radiatum (SR). Laminar profiles of evoked extracellular field potentials were recorded at 25-micron intervals along the entire dendrosomatic axis of the pyramidal cell and a one-dimensional current source-density analysis was applied. 3. Suprathreshold stimulation of the alveus evoked an antidromic population spike response and current sink with the shortest peak latency in stratum pyramidale or proximal stratum oriens. A biphasic positive/negative potential associated with a current source/sink was recorded in dendritic regions, with both components increasing in peak latency with distance from the border of stratum pyramidale. 4. Suprathreshold stimulation of SO or SR evoked a population spike response superimposed upon the underlying synaptic depolarization at all levels of the dendrosomatic axis. The shortest latency population spike and current sink were recorded in stratum pyramidale or proximal stratum oriens. In dendritic regions, a biphasic positive/negative potential and current source/sink conducted with increasing latency from the border of stratum pyramidale. 5. A direct comparison of alvear- and SR-evoked responses revealed a basic similarity in population spike potentials and associated sink/source relationships at both the somatic and dendritic level and a similar shift in peak latency of spike components along the pyramidal cell axis. 6. It is concluded that the initial site for generation of a spike along the dendrosomatic axis of the pyramidal cell following antidromic or orthodromic stimulation is in the region of the cell body layer (soma or axon hillock). Action-potential discharge in dendritic regions then occurs as the result of a subsequent retrograde spike invasion of basal and apical dendritic arborizations.

Extracellular fields influence transmembrane potentials and synchronization of hippocampal neuronal activity.

The influence of extracellular fields on the transmembrane potential (TMP) of CA1 pyramidal neurons was investigated following both ortho- and antidromic stimulation in the in vitro hippocampal slice preparation. A short latency negative deflection on the intracellular potential coincided with the falling phase of the extracellular population spike. Subtraction of extracellular field potentials from ground referenced intracellular records revealed a sharp depolarizing wave of the TMP superimposed upon the underlying synaptic potential. This graded depolarization was capable of discharging CA1 cells and displayed a parallel shift in latency and amplitude with the extracellular population spike. A similar depolarizing wave was associated with the antidromically evoked population spike which persisted following blockade of synaptic activity. Finally, multiple population spike activity similar to that observed during epileptiform discharge was associated with repetitive depolarizing waves of the TMP. These data suggest that extracellular field potentials can ephaptically discharge CA1 neurons and may play a role in recruitment and synchronization of neuronal activity in the hippocampus.

Ephaptic interactions contribute to paired pulse and frequency potentiation of hippocampal field potentials.

The contribution of ephaptic interactions to potentiation of the hippocampal CA1 extracellular population spike during paired pulse or frequency stimulation of stratum radiatum (SR) inputs was investigated using the in vitro hippocampal slice preparation. Records of the transmembrane potential revealed a depolarizing wave with an amplitude and latency that varied directly with that of the extracellular population spike. Paired pulse or repetitive stimulation of SR resulted in a potentiation of the population spike amplitude and a corresponding increase in the amplitude of the TMP depolarizing wave. Action potentials generated during the stimulus train consistently arose from the peak of the depolarizing wave. It is proposed that ephaptic interactions contribute to potentiation of the extracellular population spike through recruitment of subthreshold neurons within the population during repetitive afferent stimulation.

The excitation of mammalian central neurones by amino acids.

1. The relative potencies of a number of analogues of L-glutamate as excitants of thalamic neurones in the rat have been compared. The most powerful compounds were kainate, ibotenate and (+/-)cis-1-amino-1,3-dicarboxycyclopentane. The D- and L-isomers of glutamate and aspartate were also compared. Whereas D-glutamate is approximately one-half as active as the L-form, D-aspartate is more potent than L-aspartate. 2. Computer analysis has indicated that ibotenate and cis-1-amino-1,3-dicarboxy-cyclopentane have relatively fixed and similar C alpha-N, Comega-N and C alpha-Comega interatomic distances which can also be achieved by glutamate in certain conformations of the molecule, but not by aspartate. 3. Parallel examination of the antagonists glutamate diethylester and D-alpha-aminoadipate has shown that the former preferentially reduces L-glutamate effects while the latter blocks the actions of other amino acid excitants more readily than those of L-glutamate. 4. The evidence is consistent with the hypothesis that at least two populations of neuronal receptors for the excitatory amino acids exist.

Mechanisms of excitation and inhibition in the nigrostriatal system.

The extracellular responses of neurones in the neostriatum following single pulse stimulation of the substantia nigra were investigated in urethane anaesthetized rats. Low intensity stimulation (less than 10 V) evoked single large amplitude spikes while higher intensities (10-20 V) elicit a high frequency burst of small amplitude spikes or waves. When spontaneous or glutamate-induced large spikes are recorded, nigral stimulation causes their inhibition coincidentally with the development of a burst. If the burst is prevented, the inhibitory response disappears. Both the nigral evoked inhibition and burst response are unaffected by iontophoretically or systemically administered antonists of dopamine or by chemical lesions of the dopamine-containing nigral neurones. The monosynaptic activation of large amplitude striatal neurones, which could also be identified antidromically by stimulation of the globus pallidus, was reversibly blocked by dopamine antagonists. It is concluded (a) that the burst responses are induced through the antidromic excitation of striatonigral axons within the striatum; (b) that the striatal neurones thus activated are inhibitory interneurones and (c) that the dopamine-containing neurones of the nigra make excitatory synaptic contact with a population of striatal output cells, some of which at least project to the globus pallidus.