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The Richard F. Caris Mirror Lab at the University of Arizona continues the production of 8.4 m lightweight honeycomb segments for the primary mirror of the Giant Magellan Telescope. GMT will have a center segment surrounded by six identical off-axis segments, plus an additional off-axis segment to allow continuous operation as segments are removed for coating. Production highlights of the last two years include the spin-casting of Segment 5, preliminary polishing of Segment 2, and completion of the rear surface processing for Segments 3 and 4. We completed a preliminary design of the significant modifications of the test systems required for Segment 4, the center segment. We finished an upgrade of the 8.4 m polishing machine; both the upgrade and experience gained with Segment 1 have contributed to much faster polishing convergence for Segment 2. Prior to polishing Segment 2, we verified the stability and accuracy of the measurement systems by re-measuring Segment 1, achieving good agreement among multiple independent tests as well as good agreement with the original acceptance tests of Segment 1.

Mitotically active, growth-arrested cells and proliferatively senescent cultures of human fetal lung fibroblasts (WI-38) were exposed to six different oxygen tensions for various lengths of time and then analyzed to determine the responses of their antioxidant defense system. Glutathione (GSH) concentration increased as a function of ambient oxygen tension in early passage cultures; the effect was larger in exponentially growing cultures than in those in a state of contact-inhibited growth arrest, but was absent in senescent cells. Conversely, the activity of glutathione disulfide reductase was greater in growth-arrested cultures than in mitotically active cells irrespective of oxygen tension. Glucose-6-phosphate dehydrogenase was lowest in log-phase cells exposed to different oxygen tensions for 24 h and in senescent cells. Both hypoxia and hyperoxia depressed selenium-dependent glutathione peroxidase activity in early passage cultures, while the activity of the enzyme progressively declined with increasing oxygen in senescent cells. The GSH S-transferase activity was unresponsive to changes in ambient oxygen tension in either young or senescent cultures. Manganese-containing superoxide dismutase (MnSOD) activity was unaffected by oxygen tension, but was elevated in young confluent cultures as compared with cultures in log-phase growth. MnSOD activity was significantly higher in senescent cultures than in early passage cultures and was also responsive to increased oxygen tension in senescent cultures. Copper–zinc-containing superoxide dismutases activity was not affected by oxygen tension or the passage of time, but it declined in senescent cultures.

We present the first scientific images obtained with a deformable secondary mirror adaptive optics (AO) system. We utilized the 6.5 m Multiple Mirror Telescope adaptive optics system to produce high-resolution (FWHM ¼ 0>07) near-infrared (1.6 lm) images of the young (� 1 Myr) Orion Trapezium � 1 Ori cluster members. A combination of high spatial resolution and high signal-to-noise ratio allowed the positions of these stars to be measured to within � 0>003 accuracies. We also present slightly lower resolution (FWHM � 0>085) images from Gemini with the Hokupa‘a AO system as well. Including previous speckle data from Weigelt et al., we analyze a 6 yr baseline of high-resolution observations of this cluster. Over this baseline we are sensitive to relative proper motions of only � 0>002 yr � 1 (4.2 km s � 1 at 450 pc). At such sensitivities we detect orbital motion in the very tight � 1 Ori B2-B3 (52 AU separation) and � 1 Ori A1-A2 (94 AU separation) systems. The relative velocity in the � 1 Ori B2-B3 system is 4:2 � 2: 1k m s � 1 . We observe 16:5 � 5: 7k m s � 1 of relative motion in the � 1 Ori A1-A2 system. These velocities are consistent with those independently observed by Schertl et al. with speckle interferometry, giving us confidence that these very small (� 0>002 yr � 1 ) orbital motions are real. All five members of the � 1 Ori B system appear likely gravitationally bound (B2-B3 is moving at � 1.4 km s � 1 in the plane of the sky with respect to B1, where Vesc � 6k m s � 1 for the B group). The very lowest mass member of the � 1 Ori B system (B4) has K 0 � 11:66 and an estimated mass of � 0.2 M� . Very little motion (4 � 15 km s � 1 ) of B4 was detected with respect to B1 or B2; hence, B4 is possibly part of the � 1 Ori B group. We suspect that if this very low mass member is physically associated, it most likely is in an unstable (nonhierarchical) orbital position and will soon be ejected from the group. The � 1 Ori B system appears to be a good example of a star formation ‘‘ minicluster,’’ which may eject the lowest mass members of the cluster in the near future. This ‘‘ ejection ’’ process could play a major role in the formation of low-mass stars and brown dwarfs. Subject headings: binaries: general — instrumentation: adaptive optics — stars: evolution — stars: formation — stars: low-mass, brown dwarfs On-line material: color figures

Basal antioxidant defense levels are often aberrant in tumor cells; however, less attention has been given to differences in the way that normal and transformed cells respond to changes in oxidative stress. This study evaluated differences in the responses of various normal and transformed cell lines to different oxygen tensions. Exposure to hyperoxia generally failed to induce either the activity of GSH peroxidase (GPx) or the manganese-containing form of superoxide dismutase (MnSOD) after 48 h, although at 605 mm Hg oxygen, small inductions of MnSOD activity were observed in adult lung fibroblasts and amelanotic melanoma. Exposure to 605 mm Hg O2 for 48 h was inhibitory to GPx activity. MnSOD activity was strongly induced in virally transformed WI-38 cells by treatment with the herbicide paraquat or inhibition of GSH synthesis with BSO. In normal cells GSH concentration was proportional to ambient oxygen tension. Tumor cells exhibited greater GSH concentrations at low oxygen tensions than normal cells but were unable to increase GSH in response to elevation of oxygen tension. These results reveal differences in tumor and normal cell responses to changes in ambient oxygen tension and show that MnSOD activity is inducible when an appropriate stimulus is applied.

Neuropeptides are released into the extracellular space from large secretory granules. In order to reach their release sites, these granules are translocated on microtubules and thought to interact with filamentous actin as they approach the cell membrane. We have used a green fluorescent protein-tagged neuropeptide prohormone (prepro-orphanin FQ) to visualize vesicle trafficking dynamics in NS20Y cells and cultures of primary hippocampal neurons. We found that the majority of secretory granules were mobile and accumulated at both the tips of neurites as well as other apparently specialized cellular sites. We also used live-cell imaging to test the notion that peptidergic vesicle mobility was regulated by secretagogues. We show that treatment with forskolin appeared to increase vesicle rates of speed, while depolarization with high K+ had no effect, even though both treatments stimulated neuropeptide secretion. In cultured hippocampal neurons the green fluorescent protein-tagged secretory vesicles were routed to both dendrites and axons, indicating that peptidergic vesicle transport was not polarized. Basal peptidergic vesicle mobility rates in hippocampal neurons were the same as those in NS20Y cells. Taken together, these studies suggest that secretory vesicle mobility is regulated by specific classes of secretagogues and that neuropeptide containing secretory vesicles may be released from dendritic structures.

Segment production for the Giant Magellan Telescope is well underway, with the off-axis Segment 1 completed, off-axis Segments 2 and 3 already cast, and mold construction in progress for the casting of Segment 4, the center segment. All equipment and techniques required for segment fabrication and testing have been demonstrated in the manufacture of Segment 1. The equipment includes a 28 m test tower that incorporates four independent measurements of the segment's figure and geometry. The interferometric test uses a large asymmetric null corrector with three elements including a 3.75 m spherical mirror and a computer-generated hologram. For independent verification of the large-scale segment shape, we use a scanning pentaprism test that exploits the natural geometry of the telescope to focus collimated light to a point. The Software Configurable Optical Test System, loosely based on the Hartmann test, measures slope errors to submicroradian accuracy at high resolution over the full aperture. An enhanced laser tracker system guides the figuring through grinding and initial polishing. All measurements agree within the expected uncertainties, including three independent measurements of radius of curvature that agree within 0.3 mm. Segment 1 was polished using a 1.2 m stressed lap for smoothing and large-scale figuring, and a set of smaller passive rigid-conformal laps on an orbital polisher for deterministic small-scale figuring. For the remaining segments, the Mirror Lab is building a smaller, orbital stressed lap to combine the smoothing capability with deterministic figuring.

The Steward Observatory Mirror Lab is nearing completion of the combined primary and tertiary mirrors of the Large Synoptic Survey Telescope. Fabrication of the combined mirror requires simulation of an active-optics correction that affects both mirror surfaces in a coordinated way. As is common for large mirrors, the specification allows correction of large-scale figure errors by a simulated bending of the substrate with the 156 mirror support actuators. Any bending affects both mirrors, so this active-optics correction is constrained by the requirement of bending the substrate so both mirrors meet their figure specifications simultaneously. The starting point of the simulated correction must be measurements of both mirrors with the substrate in the same shape, i. e. the same state of mechanical and thermal stress. Polishing was carried out using a 1.2 m stressed lap for smoothing and large-scale figuring, and a set of smaller passive rigid-conformal laps on an orbital polisher for deterministic small-scale figuring. The primary mirror is accurate to about 25 nm rms surface after the active-optics correction, while work continues toward completion of the tertiary.

The hypothalamic-pituitary-adrenal (HPA) axis is regulated by stress-related excitatory inputs, and various inhibitory and negative-feedback controls by glucocorticoids and opioids, including pro-opiomelanocortin (POMC)-derived peptides. The role of POMC-derived peptides of pituitary origin in the modulation of brain POMC mRNA expression and opioid receptor binding was investigated using a line of transgenic mice that express a fusion gene composed of the pituitary expression-specific promoter region of the POMC gene driving the herpes simplex viral-1 thymidine kinase (TK). Male adult mice were treated with the antiherpes agent ganciclovir that selectively ablates cells expressing TK. Following treatment, POMC mRNA levels, measured by quantitative solution hybridization/RNase protection assays, were decreased by 48% in the pituitary of the TK+/+ mice, reflecting an expected loss of the pituitary corticotrope POMC cells. This treatment also significantly lowered pituitary beta-endorphin immunoreactivity content and plasma concentrations of corticosterone. In contrast, POMC mRNA levels were increased by 79% in the hypothalamus of the TK+/+ mice with pituitary POMC cell ablation. Binding of [(3)H]DAMGO to mu opioid receptors, as measured by quantitative autoradiography, was significantly reduced in several brain regions including the central grey, median raphe and superficial grey layer of the superior colliculus. These regions are innervated by hypothalamic POMC neurones. No significant differences in binding to either kappa or delta opioid receptors were found in the brain regions studied. These results suggest that POMC-derived peptides of pituitary origin may exert a tonic negative-feedback effect on hypothalamic POMC neurones. In turn, the downregulation of central mu opioid receptors in this model may be mediated through a mechanism related to hypothalamic POMC overexpression.

Mechanical overloading of cartilage has been implicated in the initiation and progression of osteoarthrosis. Our objectives were to identify threshold levels of strain rate and peak stress at which sub-impact loads could induce cartilage matrix damage and chondrocyte injury in bovine osteochondral explants and to explore relationships between matrix damage, spatial patterns of cell injury, and applied loads. Single sub-impact loads characterized by a constant strain rate between 3×10−5 and 0.7 s −1 to a peak stress between 3.5 and 14 MPa were applied, after which explants were maintained in culture for four days. At the higher strain rates, matrix mechanical failure (tissue cracks) and cell deactivation were most severe near the cartilage superficial zone and were associated with sustained increased release of proteoglycan from explants. In contrast, low strain rate loading was associated with cell deactivation in the absence of visible matrix damage. Furthermore, cell activity and proteoglycan synthesis were suppressed throughout the cartilage depth, but in a radially dependent manner with the most severe effects at the center of cylindrical explants. Results highlight spatial patterns of matrix damage and cell injury which depend upon the nature of injurious loading applied. These patterns of injury may also differ in terms of their long-term implications for progression of degradative disease and possibilities for cartilage repair.

We previously devised a magnetic resonance imaging method that allows for the nondestructive and quantitative determination of glycosaminoglycan concentration in excised cartilage. The technique measures the concentration of the charged contrast agent Gd-DTPA2- (gadolinium diethylenetriamine-pentaacetic acid) equilibrated within cartilage, from which the tissue distribution of glycosaminoglycan can be calculated. The goals of our study were to determine the practicality of nondestructively monitoring glycosaminoglycan concentration in cartilage explants over a long-term culture period and to determine if glycosaminoglycan could be restored to glycosaminoglycan-depleted cartilage explants maintained in long-term culture. To meet our objectives, we harvested bovine cartilage explants, treated them initially with trypsin to reduce the glycosaminoglycan concentration, and cultured them for as long as 8 weeks. Images depicting glycosaminoglycan concentration were calculated from magnetic resonance images acquired at selected intervals during the trypsinization process and the subsequent culture period. The results indicate that gadolinium-enhanced magnetic resonance imaging can follow the reduction of glycosaminoglycan concentration over the course of enzymatic digestion and the replenishment of glycosaminoglycan over several weeks of culture and that cultured cartilage explants are capable of restoring glycosaminoglycan to 85% of its initial concentration. Of particular interest, samples cultured for 5 weeks indicated a depth dependence of glycosaminoglycan regeneration to values similar to the initial physiologic distribution. Thus, this magnetic resonance imaging method may be a very powerful means for exploring the spatial and temporal evolution of glycosaminoglycan in cartilage.

A computer model, SPRINKMOD, was tested with field data collected from a complex sprinkler irrigation system in operation. The original data collected were adjusted in order to calibrate the model for the irrigation system. Five modifications were made in the original data so that the model sensitivity could be evaluated. The model predicted the system operating point with less than 1% error, after some adjustments in the data. The relative mean standard error was 4.1% for the upstream pressure heads and 2.1% for the downstream pressure heads of all laterals analyzed together. Measurement of pressure and discharge at pump stations along with a reasonable estimate of leakage appears to be essential for simulation of old systems. Adjustments made to pipe wall thickness and pipe roughness were not as important as the adjustments made to the lateral valves closure in getting SPRINKMOD to simulate pressure heads that were close to the measured values. For long laterals, like center pivot laterals, a variable local loss coefficient for flow past a riser outlet worked better than a fixed value of 0.3.

A computer model, SPRINKMOD, was tested with field data collected from a simple sprinkler irrigation system in operation. The original data for laterals and the amount and way leakage is considered were modified to evaluate the model sensitivity. The model predicted the pump station flow rate within 2% and the pump station pressure head within 5% with the original data collected. For this irrigation system, no practical effect was observed in the system operating point by changing the lateral pipe characteristics, lateral leakage amount and distribution, lateral pipe roughness and lateral local loss coefficient for flow past a riser outlet. The amount of leakage had more effect on the model simulation of pressure heads than the way leakage was considered, localized or distributed along the laterals. The use of a variable local loss coefficient, K r, along the 350-m hand-move laterals had a negligible effect on both system operating point and distribution of pressure heads along the laterals.

Adjustment coefficients for air temperature and dew point temperature measurements at dry weather sites are determined for predicting reference evapotranspirations (ET0) for irrigated conditions. Seasonal and regional variation in changes in temperature parameters versus the ratio of effective precipitation to ET0 are analyzed. The determined adjustment coefficients are tested for validity using weather data from reference weather stations, and paired reference and dry stations. The effects of adjusting air temperature and dew point temperature for site aridity are analyzed using the FAO Penman-Monteith and the Hargreaves equations. Maximum reductions in ET0 caused by adjustments of temperature data for aridity effects was as high as 25%.

The Food and Agriculture Organization of the United Nations (FAO) version of the Penman-Monteith method (FAO-PM) has recently been proposed as the standard for estimating reference evapotranspiration (ETo). A standard, well-watered reference environment for the data used for the ETo estimate has also been proposed. At those sites providing a reference environment the average minimum air temperature (Tn) is approximately the same as average dew point temperature (Td). For this reason and because the computation of Td is plagued with errors, the FAO-PM can be modified to use Td=Tn. This modification is referred to as FAO-MOD. However, most weather stations lack the intensive input required by the FAO-PM. Few weather stations provide a standard environment. Errors in measuring the required weather variables are common. Under these conditions, equations with few weather parameters are recommended. The Hargreaves et al. (1985) method (ETH) is one of the simplest and most accurate empirical equations. The ETH m...

A software package called Command Area Decision Support Model (CADSM) was developed to estimate aggregate crop-water requirements and to study management options for irrigated areas. Daily water and salt balances are simulated for individual fields within command areas based on crop type and stage of development, field characteristics, soil properties, possible ground-water contribution, salinity level, and several queuing factors that take cultural practices into account. The water requirements for individual fields are aggregated to estimate the potential command area water requirement, then totaled to determine the potential irrigation system demand. A queuing system is used to allocate available water to command areas and fields. Average crop yield response is predicted considering root-zone water deficit, salinity concentration, and water logging. Daily weather data, including evaporation, air temperature, and reference crop evapotranspiration (ETo) can be generated using monthly statistical means an...

Effective use of water supplies via irrigation requires good methods for determining crop water needs. To implement irrigation scheduling programs requires an accurate estimate of water use by the ...

In this chapter, we present standard procedures for the culture of human cells that exhibit a finite proliferative capacity (replicative life-span). The use of a cell culture model has the advantage of providing a controlled environment to study a wide variety of cellular phenomena. It also has the inherent limitation of isolating cells from the regulatory elements that might be provided by other types of cells in vivo. Nevertheless, cell culture models have been crucial to our current understanding of mechanisms of growth, differentiation, development, and neoplasia and numerous other disease states. In this chapter we present procedures for human fibroblast culture including serumfree cultivation of cells, which is necessary when the cellular environment must be fully defined. In addition, we present procedures for the determination of replicative life-span, saturation density, and assessment of replicative capacity from labeled thymidine incorporation in fibroblasts. The methods described here have been well tested and provide highly reproducible results (1, 2).

Cartilage functionality is defined, in part, in terms of the ability of the extracellular matrix to support a mechanical load. It has been shown that such mechanical loading can influence the biological response of the chondrocytes that are embedded in the extracellular matrix. Cultured tissue explants have served as useful models for studying such chondrocyte-mediated responses to mechanical deformation. The explant paradigm facilitates the control of mechanical and biological variables that may influence cellular behavior. This chapter presents the means for assessing the biologic response of cartilage to controlled mechanical stimuli, and lays the foundation to further explore the response of cartilage to mechanical stimuli in the presence of other factors, such as cytokines and growth factors (e.g., IL-1β, TGF-β, IGF).

The primary mirror of the Giant Magellan Telescope consists of seven 8.4 m segments which are borosilicate honeycomb sandwich mirrors. Fabrication and testing of the off-axis segments is challenging and has led to a number of innovations in manufacturing technology. The polishing system includes an actively stressed lap that follows the shape of the aspheric surface, used for large-scale figuring and smoothing, and a passive "rigid conformal lap" for small-scale figuring and smoothing. Four independent measurement systems support all stages of fabrication and provide redundant measurements of all critical parameters including mirror figure, radius of curvature, off-axis distance and clocking. The first measurement uses a laser tracker to scan the surface, with external references to compensate for rigid body displacements and refractive index variations. The main optical test is a full-aperture interferometric measurement, but it requires an asymmetric null corrector with three elements, including a 3.75 m mirror and a computer-generated hologram, to compensate for the surface's 14 mm departure from the best-fit sphere. Two additional optical tests measure large-scale and small-scale structure, with some overlap. Together these measurements provide high confidence that the segments meet all requirements.

The effects of a static magnetic field (SMF) on the proliferation of various types of human cells were determined. All cultures were maintained at 37 °C throughout the experiment. SMF was generated by placing two magnets oppositely oriented on either side of a T25 flask. The flux density in the flask ranged from 35 to 120 mT. Growth curves were constructed by plotting cell number at 18 h and 4, 7, 11, and 14 days after seeding, with the 18-h point being a measure of attachment efficiency. Exposure to SMF significantly decreased initial attachment of fibroblasts and decreased subsequent growth compared to sham-exposed control. Significant effects were observed in both fetal lung (WI-38) and adult skin fibroblasts, but they were generally larger in the fetal lung fibroblast line. SMF did not affect attachment of human melanoma cells, but inhibited their growth by 20% on day 7. SMF produced no effects in a human adult stem cell line. Oxidant production increased 37% in WI-38 cells exposed to SMF (230-250 mT) during the first 18 h after seeding, when cell attachment occurs. Conversely, no elevation in oxidant levels was observed after a prolonged 5-day exposure. These results indicate that exposure to SMF has significant biological effects in some, but not all types of human cells.

Evapotranspiration coefficients and aerodynamic roughness and bulk stomatal resistance parameters were developed for predicting evapotranspiration (Et) from narrow, isolated stands of cattail and bulrush (hydrophytes) using measurements from drainage lysimeters. Peak ratios of Et from vegetation in the center of 6 m wide stands of hydrophytes to Et from alfalfa averaged 1.6 for cattails and 1.8 for bulrushes. A ratio of momentum roughness height to vegetative height of 0.3 for use in the Penman-Monteith equation best fit experimental data. Errors between prediction equations and lysimeter measurements were minimized using linear, segmented evapotranspiration coefficient curves as compared to using a fitted Penman-Monteith approach.

Diethylstilbestrol induces proliferation of Syrian hamster renal proximal tubular cells. By counting the number of cells in culture, we showed that liposomes containing superoxide dismutase or catalase suppressed diethylstilbestrol-induced proliferation, whereas empty liposomes or liposomes containing inactivated superoxide dismutase did not. Liposomes containing antioxidant enzymes did not suppress proliferation of cells in control media or of cells treated with ethinyl estradiol. In the absence of liposomes, exogenous superoxide dismutase did not suppress diethylstilbestrol-induced proliferation. The decrease in cell number when diethylstilbestrol-treated cells were treated with antioxidant enzyme-containing liposomes was not due to decreased cell viability. Results were confirmed by measuring a correlate of cell proliferation immunohistochemically, using an antibody to proliferating cell nuclear antigen. A larger proportion of diethylstilbestrol-treated cells than of control cell showed nuclear immunostaining with this antibody. The number of cells immunostained in diethylstilbestrol-treated cultures was sharply decreased by the addition of superoxide dismutase- or catalase-containing liposomes. Our studies suggest a role for active oxygen species in diethylstilbestrol-induced proliferation of cultured proximal tubular cells.

The second 8.4 m primary mirror and its active support system were delivered to the Large Binocular Telescope in September 2005. The mirror was figured to an accuracy of 15 nm rms surface after subtraction of low-order aberrations that will be controlled by the active support. The mirror was installed into its operational support cell in the lab, and support forces were optimized to produce a figure accurate to 20 nm rms surface with no synthetic correction. The mirror was polished on a new 8.4 m polishing machine that gives the Mirror Lab the capacity to process up to four 8.4 m mirrors simultaneously, with each mirror going through a sequence of stations: casting furnace, generating machine, polishing machine, and integration with its support cell. The new polishing machine has two carriages for polishing tools, allowing use of two 1.2 m stressed laps during loose-abrasive grinding and early polishing, followed by final figuring with a stressed lap and a small tool for local figuring.

Aging occurs in all aerobic multicellular species. Subcellular oxidant production can damage nucleic acids, lipids, and proteins providing a potential link to aging, but a specific mechanism by which oxidants cause the aging process has not been established. Cells contain antioxidant defenses that can mediate the deleterious effects of oxidant production. In some cases, the targets of oxidative stress may involve changes in regulatory pathways rather than direct damage to physical structures.

The objective of this study was to determine the dose–response effects of epinephrine, given by systemic intravenous infusion to the halothane-anesthetized newborn piglet, on renal blood flow, mean arterial blood pressure, and renal vascular resistance. Seven newborn piglets were acutely instrumented. A transit-time ultrasound flow probe was placed around the renal artery and a femoral arterial catheter was placed for blood pressure monitoring. Epinephrine was infused in doubling doses from 0.2 to 3.2 μg∙kg−1∙min−1. Mean arterial blood pressure increased from 54 mmHg (1 mmHg = 133.3 Pa) to an average of 96 mmHg at 3.2 μg∙kg−1∙min−1 of epinephrine. Renal blood flow increased from 165 mL∙min−1∙100 g−1 at baseline to 185 mL∙min−1∙100 g−1 at a dose of 0.2 μg∙kg−1∙min−1 and increased further at 0.4 and 0.8 μg∙kg−1∙min−1 to reach 261 mL∙min−1∙100 g−1. Renal blood flow began to fall at a dose of 3.2 μg∙kg−1∙min−1, remaining however, significantly above baseline (211 mL∙min−1∙100 g−1). Consequently, calculated renal vascular resistance fell as the dose was increased from 0.2 to 0.8 μg∙kg−1∙min−1 and then rose again at 1.6 and 3.2 μg∙kg−1∙min−1, being significantly above baseline at 3.2 μg∙kg−1∙min−1. These results demonstrate that epinephrine when given by systemic infusion to the halothane-anesthetized newborn pig is a renal vasodilator at low doses and causes renal vasoconstriction at moderate to high doses. Renal blood flow remained above baseline at all doses tested, and thus, within the dosage range tested, epinephrine infusion should not cause renal ischemia.Key words: epinephrine, kidney blood flow, piglet, renal vascular resistance.

Neuropeptides are released into the extracellular space from large secretory granules. In order to reach their release sites, these granules are translocated on microtubules and thought to interact with filamentous actin as they approach the cell membrane. We have used a green fluorescent protein-tagged neuropeptide prohormone (prepro-orphanin FQ) to visualize vesicle trafficking dynamics in NS20Y cells and cultures of primary hippocampal neurons. We found that the majority of secretory granules were mobile and accumulated at both the tips of neurites as well as other apparently specialized cellular sites. We also used live-cell imaging to test the notion that peptidergic vesicle mobility was regulated by secretagogues. We show that treatment with forskolin appeared to increase vesicle rates of speed, while depolarization with high K+ had no effect, even though both treatments stimulated neuropeptide secretion. In cultured hippocampal neurons the green fluorescent protein-tagged secretory vesicles were routed to both dendrites and axons, indicating that peptidergic vesicle transport was not polarized. Basal peptidergic vesicle mobility rates in hippocampal neurons were the same as those in NS20Y cells. Taken together, these studies suggest that secretory vesicle mobility is regulated by specific classes of secretagogues and that neuropeptide containing secretory vesicles may be released from dendritic structures.

Oxygen toxicity is believed to arise from changes in the rates at which cells generate reactive oxygen species (ROS). Sensitivity to hyperoxia has been postulated to depend on levels of antioxidant defense. Human cells obtained from fetal tissues have lower antioxidant defenses than those obtained from adult tissue. The present study was performed to determine whether the differences in fetal and adult antioxidant defense levels modulated their responses to changes in the ambient oxygen concentration. Our results demonstrate that oxygen modulates the proliferation of human fetal and adult skin fibroblasts in a similar fashion. In general, skin fibroblasts grew better at approximately 31 mm Hg, regardless of donor age. Manganese superoxide dismutase, catalase, and glutathione peroxidase activities were lower in fetal cells than in adult fibroblasts. Copper/zinc superoxide dismutase and glucose-6-phosphate dehydrogenase were similar in fetal and postnatal tissues and were unaltered appreciably by hyperoxic exposure. Glutathione concentration increased at higher oxygen tensions; however, the increase was much greater in fetal cells than in cultures derived from adult skin. These observations demonstrate that the capacity of fetal and adult cells to cope with oxidative stress, while similar, result from distinct mechanisms.

The adaptive optics (AO) system for the 6.5 m MMT conversion telescope is the first to compensate the aberrated wavefront at the telescope's secondary mirror. This approach has unique advantages in terms of optical simplicity, high throughput and low emissivity. Its realization presents many technical challenges, which have now been overcome. The deformable mirror is now characterized and accepted. It features a 1.9mm thick 640mm diameter convex aspheric mirror (manufactured at the Steward Observatory Mirror Lab), mounted on a 50 mm thick ULE reference body with 336 actuators, as well as a cluster of 168 DSP's and associated analog circuitry. A wavefront sensor with integrated CCD and lenslet array has also been completed. The complete system is now starting to produce laboratory results, which we present below. Closed loop operation is tested under an auto-collimation illumination system that reflects aberrated artificial starlight from the convex secondary.© (2002) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

The hypothalamic-pituitary-adrenal (HPA) axis is regulated by stress-related excitatory inputs, and various inhibitory and negative-feedback controls by glucocorticoids and opioids, including pro-opiomelanocortin (POMC)-derived peptides. The role of POMC-derived peptides of pituitary origin in the modulation of brain POMC mRNA expression and opioid receptor binding was investigated using a line of transgenic mice that express a fusion gene composed of the pituitary expression-specific promoter region of the POMC gene driving the herpes simplex viral-1 thymidine kinase (TK). Male adult mice were treated with the antiherpes agent ganciclovir that selectively ablates cells expressing TK. Following treatment, POMC mRNA levels, measured by quantitative solution hybridization/RNase protection assays, were decreased by 48% in the pituitary of the TK+/+ mice, reflecting an expected loss of the pituitary corticotrope POMC cells. This treatment also significantly lowered pituitary beta-endorphin immunoreactivity content and plasma concentrations of corticosterone. In contrast, POMC mRNA levels were increased by 79% in the hypothalamus of the TK+/+ mice with pituitary POMC cell ablation. Binding of [(3)H]DAMGO to mu opioid receptors, as measured by quantitative autoradiography, was significantly reduced in several brain regions including the central grey, median raphe and superficial grey layer of the superior colliculus. These regions are innervated by hypothalamic POMC neurones. No significant differences in binding to either kappa or delta opioid receptors were found in the brain regions studied. These results suggest that POMC-derived peptides of pituitary origin may exert a tonic negative-feedback effect on hypothalamic POMC neurones. In turn, the downregulation of central mu opioid receptors in this model may be mediated through a mechanism related to hypothalamic POMC overexpression.

We describe the optical fabrication and the active support system of the 6.5 m f/1.25 primary mirror for the first Magellan telescope. Figuring was performed with a 1.2 m stressed lap, which bends under active control to match the local curvature of the optical surface, and small passive tools. The figure was measured with IR and visible interferometers, using refractive null lenses to compensate 810 microns of aspheric departure. After subtraction of Seidel astigmatism and spherical aberration, the finished mirror is accurate to 14 nm rms surface and has an encircled energy of 80% in 0.06' diameter at 500 nm. The mirror was integrated with its active support system in the laboratory, and support forces were adjusted to optimize the figure. The optimization was performed by singular value decomposition of the influence functions into normal bending modes. Using the first 20 modes and a maximum correction force of 46 N, the surface accuracy is 24 nm rms with 80% of the light in 0.11' diameter.

Objective: To test the hypothesis that prolonged alkalosis decreases cardiac output and, furthermore, exacerbates hypoxic pulmonary vasoconstriction, as respiratory alkalosis is frequently induced as a therapy for persistent pulmonary hypertension of the newborn despite a lack of controlled evidence of improved outcomes. Potential adverse effects of prolonged alkalosis have been demonstrated.¶Method: Two groups (control, n = 6, and hypocapnic alkalosis, n = 6) of 1–3 day old fentanyl-anesthetized, vecuronium-paralyzed piglets were instrumented to measure cardiac index (CI) and mean systemic (MAP) and pulmonary (PAP) arterial pressures. Baseline values were recorded. Alveolar hypoxia was then induced to achieve an arterial oxygen saturation of between 50 and 60 % for 15 min. Respiratory alkalosis was then induced, by increasing ventilation to achieve a pH between 7.55–7.60, and was continued for 240 min. Inspired carbon dioxide was used with hyperventilation in the control group to maintain pressure of arterial carbon dioxide (PaCO2) at 35–45 mmHg and pH of 7.35–7.45. Hypoxia was induced again at 15 and 240 min. Pulmonary and systemic vascular resistances (PVR and SVR) were calculated.¶Results: Prolonged alkalosis led to a significant and progressive fall in mean MAP from 61 (SD 7) mmHg at the start of the study falling to 50 (SD 6.9, p = 0.043), with no effect on CI. Calculated SVR decreased (0.45 SD 0.03 vs 0.36 SD 0.05). There were no statistically significant changes in any of the variables in the control group. Neither acute nor prolonged respiratory alkalosis had a significant effect on hypoxic pulmonary vasoconstriction.¶Conclusions: Prolonged hyperventilation leads to systemic hypotension, however it does not exacerbate hypoxic pulmonary vasoconstriction.

1. The actions of the neuropeptide nociceptin, the putative nociceptin receptor antagonist [Phe1psi(CH(2)-NH)Gly(2)]-nociceptin-(1 - 13)NH(2) (Phe(1)psi-nociceptin(1 - 13)) and the putative nociceptin precursor products nocistatin (rat prepronociceptin(125 - 132)) and rat prepronociceptin(154 - 181) were examined on membrane properties of rat locus coeruleus (LC) neurons using whole cell patch clamp techniques. 2. Nociceptin inhibited I(Ba) in all LC neurons, (pD(2) of 8.9, maximum inhibition 50%). The inhibition of I(Ba) by nociceptin was associated with slowing of the activation of I(Ba) and could be significantly reversed by a strong depolarizing prepulse. Phe(1)psi-nociceptin(1 - 13) also inhibited I(Ba) in LC neurons (notional pD(2) of 7.6, maximum inhibition 18%). Application of Phe(1)psi-nociceptin(1 - 13) (1 microM) significantly occluded the subsequent effects of a co-application of nociceptin (3 nM) on I(Ba). 3. As previously reported for nociceptin, Phe(1)psi-nociceptin(1 - 13) caused an outward current in LC neurons voltage clamped at -60 mV (pD(2) of 7.1, maximum current 50% of that of methionine enkephalin, 10 microM). The Phe(1)psi-nociceptin(1 - 13) induced current reversed polarity at -112 mV and exhibited pronounced inward rectification. Phe(1)psi-nociceptin(1 - 13) (1 microM) reversibly inhibited the current caused by nociceptin (300 nM) by 30%. 4. Neither nocistatin nor rat prepronociceptin(154 - 181) inhibited I(Ba) in LC neurons, or prevented the subsequent inhibition by nociceptin. Neither nocistatin or prepronociceptin(154 - 181) affected the membrane properties of LC neurons. 5. This study demonstrates that nociceptin modulates somatic I(Ba) in rat LC neurons. The putative ORL1 antagonist Phe(1)psi-nociceptin(1 - 13) exhibited partial agonist activity at inhibiting I(Ba) and opening K(+) channels in LC. Other putative nociceptin precursor products were without effect on LC cells.

The viscosity of some polymeric fluids increases significantly when measured at pressures above atmospheric pressure. If this phenomenon is not taken into account for example when using CAD programmes to design injection moulding tools, then consequences such as short mouldings and inferior mechanical properties may be obtained, resulting in a high scrap rate and the need for expensive tool modifications. A simple new method of measuring the pressure coefficient of viscosity using a tandem-die technique with a modified capillary extrusion rheometer is described.

We describe the optical fabrication and testing of the 6.5 m f/1.25 primary mirror for the first Magellan telescope. Figuring was performed with a 1.2m stressed lap, which bends under active control to match the local curvature of the optical surface, and a variety of small passive tools. The figure was measured with RI and visible interferometers, using refractive null correctors to compensate 810 microns of aspheric departure. After subtraction of Seidel astigmatism and spherical aberration, the finished mirror is accurate to 14 nm rms surface error, and has an encircled energy of 80 percent in 0.06 inch diameter at 500 nm.

The activities and mRNA abundances of enzymes that regulate the rate of electron flow through the electron transport chain (ETC), including NADH dehydrogenase, succinate dehydrogenase, and cytochrome c oxidase, were examined in young and senescent fetal lung fibroblasts (WI-38). We also determined the activities and mRNA abundances of antioxidant defenses including superoxide dismutase, catalase, and glutathione peroxidase. We confirmed our previous report of a senescence-related increase in the abundance of ND4, a mitochondrially encoded subunit of NADH dehydrogenase. The activities of cytochrome c oxidase and NADH dehydrogenase were also elevated in senescent cultures. No differences were observed in the mRNA abundances of COX-1, a mitochondrially encoded subunit of cytochrome c oxidase or of nuclearly encoded subunits of various electron transport components (SD, COX-4, and ND 51). Lucigenin-detected chemiluminescence and H2O2 generation were both elevated in senescent cells. Catalase activity was also elevated in senescent fibroblasts. However, no differences in catalase mRNA abundance were observed. A small decrease in GSH peroxidase (GPx) mRNA abundance was observed in senescent cells. No other changes in the activities or mRNA abundances of any of the antioxidant defenses were observed in early and late passage cultures. The relationships between oxidant generation, mitochondrial enzyme activities, and antioxidant defense observed during proliferative senescence are dissimilar to those detected between fetal and postnatal fibroblasts as well as those found between fibroblast lines obtained from young and old individuals. The relevance of the differences between these models is discussed.

EPC-1 (early population doubling level cDNA-1) is a quiescence-specific gene expressed at high levels by early passage WI-38 fibroblasts under conditions of either density-dependent growth arrest or serum deprivation. Late passage WI-38 cells lose the ability to express EPC-1 under all conditions tested. The decline in EPC-1 mRNA is gradual during the replicative life span and correlates inversely with the population doubling level (PDL) of the cells. The objective of this study was to determine whether the decline in EPC-7 mRNA abundance observed during proliferative senescence also occurs in cultures derived from donors of different ages. To address this question, we examined the abundance of EPC-1 mRNA in 28 skin fibroblast lines established from healthy donors of different ages ranging from 12 fetal weeks to 94 years. EPC-1 expression was measured, under conditions of growth arrest, prior to the end of the replicative life span of the cultures. Despite some variability in steady-state transcript levels among the cell lines, EPC-1 expression was significantly lower in cells derived from the fetal donor group (12-20 gestational weeks) than in cells derived from adult donors. An in vitro age-dependent decline in EPC-1 expression was observed in all the skin lines examined, independent of donor age; however, no significant difference was observed between the young adult donor group (17-33 years) and the old adult donor group (78-94 years). Thus, expression of EPC-1 is linked to the replicative age of the cells and whether the cells are derived from fetal skin or adult skin. In adults, EPC-1 expression is independent of donor age.

We describe the fabrication of the 6.5 m f/1.25 primary mirrors for the Multiple Mirror Telescope Conversion and the first Magellan telescope. Figuring was performed with a 1.2 m stressed lap-which bends under active control to match the local curvature of the optical surface-and small passive tools. The figure was measured with IR and visible interferometers, using refractive null correctors to compensate 810 microns of aspheric departure. We present the final measurements as surface maps, synthetic interferograms and synthetic images.

The primary mirror for the Multiple Mirror Telescope Conversion is the first 6.5 m honeycomb sandwich mirror cast and polished by the Steward Observatory Mirror Lab. We describe the optical fabrication and testing of the f/1.25 paraboloid, and present the final measurements of figure accuracy and inferred image quality. Figuring was performed with a 1.2 m stressed lap--which bends under active control to match the local curvature of the optical surface--and a variety of small passive tools. The mirror was pressurized to compensate for polishing loads and thereby eliminate print-through of the honeycomb structure. The net result is a smoother surface on scales of 5 - 20 cm than has been achieved on previous honeycomb sandwich mirrors. The figure was measured with IR and visible interferometers, using refractive null correctors to compensate 810 microns of aspheric departure. The final measurements were used to calculate synthetic stellar images in a variety of seeing conditions.

We have examined the effects of three structurally distinct antioxidants (N-acetylcysteine [NAC], Trolox C [a water-soluble vitamin E derivative], and nordihydroguaiaretic acid [NGA]) on the expression of the c-fos gene over a 2-hour period. Determination of cellular glutathione concentration (the primary determinant of the cellular redox state) over the same time-course verifies that all the compounds studied cause an increase in cellular reduction potential. The level of c-fos messenger RNA increased rapidly in response to micromolar concentrations of these compounds, reaching a peak in 30-60 minutes. Induction of c-fos expression by these antioxidants is at least partly due to an increase in transcription, as determined by nuclear run-on assay. Down regulation of protein kinase C (PKC) by pretreatment for 24 hours with 500 nm PMA prevents induction by subsequent stimulation with either PMA or NGA. NAC induction of c-fos is unaffected by PMA pretreatment, while Trolox C superinduced c-fos following PMA pretreatment. None of these treatments stimulated translocation of PKC-alpha from the cytosol to the membrane. These results suggest that increasing the intracellular reducing potential induces c-fos expression through multiple pathways.

We have examined the activities and mRNA abundance of two hydrogen peroxide metabolizing enzymes (glutathione peroxidase and catalase), glutathione concentration, and the activities of several enzymes that influence glutathione concentration, including glutathione reductase (GR), glucose-6-phosphate dehydrogenase (G-6-PD), and gamma-glutamylcysteine synthetase (gamma-GCS), in 29 skin fibroblast lines derived from donors ranging in age from 14 gestational weeks to 94 years of age. H2O2 metabolizing enzyme activities and mRNA abundances were greater in skin fibroblast cultures established from postnatal donors than in fetally derived cultures. There were no significant differences in either of these parameters in cell lines established from postnatal donors of different ages. Total glutathione concentration decreased with age, but GR activity appeared to be unaffected by age. In order to estimate the ability of the cultures to produce NADPH (an important component of cellular redox status and a cofactor for GR), we determined glucose-6-phosphate dehydrogenase activity and mRNA abundance. We were unable to directly measure gamma-GCS activity or mRNA abundance in any of the skin lines or in fetal lung fibroblast; however, we were able to indirectly demonstrate the presence of this enzyme by stimulating fetal lung fibroblasts with H2O2 following treatment with L-buthionine-S,R-sulfoximine (BSO), an inhibitor of gamma-GCS activity. These results show that some, but not all, age-associated differences in antioxidant defense levels are maintained in a culture environment and are consistent with the hypothesis that developmental stages of life are associated with lower antioxidant defense levels than are present in postnatal phases of life.

We have determined the activities, protein, and mRNA abundances as well as the level of transcriptional activation of two intracellular forms of the free radical metabolizing enzyme superoxide dismutase in 29 human skin fibroblast lines established from donors of different ages. SOD-1 (a copper and zinc containing form of SOD) and SOD-2 (a manganese containing form of the enzyme) activities were both observed to be significantly lower in cell lines derived from fetal skin than in lines established from postnatal skin (ages 17-94 years). The percent of total activity contributed by SOD-1 decreased in an age-associated manner from approximately 50% in the fetal lines to less than 20% in lines established from old tissue donors. All of the cell lines were screened to exclude the possibility that they contained a polymorphism known to influence SOD-2 activity. Northern blot analysis revealed three SOD-1 mRNA transcripts that were 0.5, 0.7, and 1.9 kb in length. Although SOD-1 protein abundance was lower in fetal lines than in lines derived from postnatal donors, SOD-1 mRNA abundance did not differ between fetal cells and cell lines derived from young donors. SOD-2 protein abundance and mRNA abundance were both significantly lower in fetal lines than in postnatal lines. No postnatal age-dependent differences were observed in any of the SOD-2 parameters examined. Nuclear run-on analysis revealed that fetal cell lines exhibited a lower level of transcriptional initiation for SOD-1 than postnatal lines. The transcription of SOD-2 was readily detected in postnatal lines, but undetectable in fetal lines. These results are consistent with multiple levels of control for SOD-1 expression and with a strong transcriptional influence on SOD-2 expression.

We have produced and characterized lines of transgenic mice expressing a fusion gene composed of the pituitary expression-specific promoter region of the POMC gene, driving the herpes simplex viral-1 thymidine kinase. Adult mice were treated with the antiherpes agent ganciclovir at 70 mg/kg body weight (ip, twice daily for 10-12 days). Approximately 98% of the pituitary intermediate lobe melanotropes and anterior lobe corticotropes were ablated as determined by immunocytochemistry and RIA specific for the POMC-derived peptides, ACTH, beta-endorphin, and alpha-MSH. The number of lactotropes, somatotropes, thyrotropes, and gonadotropes was not altered compared with controls, indicating that in the adult pituitary, POMC products are not required to maintain the distribution of cell types. As expected, plasma corticosterone levels were substantially decreased after POMC cell ablation. In situ hybridization studies showed that the mouse ACTH receptor was expressed uniformly throughout the adrenal cortex, and RNase protection assays revealed that the ACTH receptor mRNA decreased after pituitary POMC cell ablation. Additionally, RNase protection assays showed that pituitary POMC cell ablation resulted in the decrease of adrenal p450c11 beta transcripts while p450c11AS (aldosterone synthase) mRNA levels remained constant. These data demonstrate differential regulation of steroid pathway-specific enzymes by POMC products. Our results also suggest that the thymidine kinase cell obliteration technique may not be dependent on cell division as a prerequisite for cytotoxicity, thus supporting the idea that targeted molecular ablation using cell- and tissue-specific promoter sequences to drive viral thymidine kinase expression can be refined further to study other nonmitotic cells.

Targeted tumorigenesis, using the POMC gene promoter ligated to the simian virus 40 large T antigen, generated transgenic mice with massive tumors of the intermediate lobe (IL) of the pituitary. Inoculation of nude mice with the IL tumor cells resulted in very large secondary tumors. As the IL from several species produces a potent PRL-releasing factor (PRF), it was of interest to determine whether IL tumors from these mice also contain PRF. The objectives were to 1) measure serum PRL levels in mice with IL tumors, 2) determine whether these tumors contain PRF and examine its chromatographic properties, and 3) analyze whether this PRF is related to POMC, its derivatives, or other PRL secretagogues. Serum PRL levels were 5- to 6-fold higher in transgenic than in control mice. Primary and secondary IL tumors were acid extracted and successively fractionated using Sephadex G-100 gel filtration and reverse phase and gel permeation HPLC. PRF activity was determined using short term incubation of tissue extracts or column fractions with GH3 cells. Crude tumor extracts exhibited a strong and dose-dependent PRF activity. Upon chromatography, the PRF activity from either primary or secondary tumors resolved into two classes of compounds: a big PRF with an estimated mol wt of 70-80 kilodaltons and two small, very hydrophobic peptides. The elution profiles of the three PRFs differed from those of beta-endorphin, alpha MSH, beta MSH, ACTH, TRH, oxytocin, angiotensin II, vasoactive intestinal polypeptide, or corticotropin-like intermediate peptide. In summary, we have identified an animal model with IL tumors that has hyperprolactinemia and overproduces PRF. Two classes of PRFs, big and small, were resolved which differ from POMC derivatives and known regulators of PRL release. These data suggest that PRF is produced by melanotrophs, but is not a product of the POMC gene. The IL tumors should provide an excellent source for the purification and structural elucidation of PRFs.

Mice harboring a transgene composed of proopiomelanocortin (POMC) gene promoter sequences (nucleotides -706 to +64) ligated to the simian virus (SV) 40 early gene encoding large T antigen developed large POMC-expressing pituitary tumors. Histologically the tumors arose from the intermediate lobe, contained nuclear SV40 T antigen and POMC peptides, but did not express other pituitary hormones. POMC processing in the pituitary tumors was indistinguishable from normal mouse intermediate lobe melanotrophs and was characterized by high proportions of acetylated and carboxyl-terminal shortened beta-endorphins, and amino-terminal acetylated alpha-melanocyte-stimulating hormone, and virtually no adrenocorticotropic hormone (ACTH)(1-39), beta-lipotropin, or POMC. The tumors contained abundant levels of mRNA for the prohormone convertase PC2 and undetectable levels of PC1. Normal mouse neurointermediate lobe also has a high ratio of PC2/PC1 expression that is distinct from the relative abundance of PC1 in anterior lobe and AtT-20 corticotroph cells. In contrast, extracts from tumors transplanted subcutaneously in nude mice contained predominantly nonacetylated forms of beta-endorphin(1-31) and -(1-27), very little ACTH(1-39), almost no corticotropin-like intermediate peptide or alpha-melanocyte-stimulating hormone, and higher proportions of intact POMC. Surprisingly, despite the less efficient proteolytic cleavage, a transplanted tumor expressed both PC1 and PC2. These studies are the first biochemical documentation of a melanotroph pituitary tumor in a rodent species and provide a new model for the investigation of pituitary oncogenesis and the molecular basis of tissue-specific prohormone post-translational processing.

Astrocytes have previously been shown to respond to cytokines such as interleukin-1 beta, tumor necrosis factor-alpha, and gamma-interferon from multiple sources including microglia and astrocytes. Recently, astrocytes have also been shown to express the opioid precursor gene proenkephalin and proenkephalin-derived peptides. The objectives of the current study were to determine if immune cytokines regulate proenkephalin gene expression in primary cultures of neonatal rat cerebral astrocytes. Northern analysis of RNA from primary cultures of neonatal rat cerebral astrocytes indicated that proenkephalin transcript levels were decreased by approximately 50% with gamma-interferon treatment and increased approximately 100% by treatment with both tumor necrosis factor-alpha and interleukin-1 beta relative to untreated controls. Tumor necrosis factor-alpha treatment was unable to reverse the inhibitory effect of gamma-interferon pretreatment on proenkephalin messenger RNA levels in the astrocytes. In contrast, expression of the constitutively expressed glutamine synthetase gene was not altered by either tumor necrosis factor-alpha or gamma-interferon treatment. These cytokines also regulate the secretion of proenkephalin-derived peptides from astrocytes. The levels of immunoreactive Met-enkephalin-Arg6-Phe7 were increased by approximately 50% with tumor necrosis factor-alpha and decreased by approximately 40% with gamma-interferon relative to untreated controls. Tumor necrosis factor-alpha was again unable to reverse the inhibitory effect of gamma-interferon pretreatment on the secretion of proenkephalin-derived peptides. These results provide additional support for the hypothesis that rapidly proliferating astrocytes may serve an important and pivotal role in mediating the bi-directional neuroimmune interactions during central nervous system disease, infection, or trauma.

The arousal responses after occlusion of the airway at the mid-trachea were compared with the responses after occlusion of the airway in a face mask in chronically instrumented 3- to 5-day-old piglets. For each site of occlusion arousal latency was significantly longer from active sleep than from quiet sleep. There was a significant increase in the frequency of early arousals after face mask occlusions compared with tracheal occlusions in both sleep states. During quiet sleep the frequency of arousal by 1 s after occlusion was 0.55 with face mask occlusions compared with 0.28 with tracheal occlusion (P less than 0.01). During active sleep the frequency of arousal by 3 s after a face mask occlusion was 0.32 compared with 0.08 after tracheal occlusion (P less than 0.05). Arousal from quiet sleep occurred before changes in arterial oxygen saturation. During active sleep mean saturation at arousal was not different between face mask and tracheal occlusions. Exposure of the upper airway to the pressures generated during airway occlusions results in earlier arousal in both quiet and active sleep, indicating a potential role for upper airway mechanoreceptors in initiating arousal in the newborn piglet.