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At least 37 records · Page 2

Intracellular pH changes in human aortic smooth muscle cells in response to fluid shear stress

The smooth muscle cell (SMC) layers of human arteries may be exposed to blood flow after endothelium denudation, for example, following balloon angioplasty treatment. These SMCs are also constantly subjected to pressure driven transmural fluid flow. Flow-induced shear stress can alter SMC growth and metabolism. Signal transduction mechanisms involved in these flow effects on SMCs are still poorly understood. In this work, the hypothesis that shear stress alters the intracellular pH (pHi) of SMC is examined. When exposed to venous and arterial levels of shear stress, human aortic smooth muscle cells (hASMC) undergo alkalinization. The alkalinization plateau persisted even after 20 min of cell exposure to flow. Addition of amiloride (10 micromoles) or its 5-(N-ethyl-N-isopropyl) analog (EIPA, 10 micromoles), both Na+/H+ exchanger inhibitors, attenuated intracellular alkalinization, suggesting the involvement of the Na+/H+ exchanger in this response. The same concentrations of these inhibitors did not show an effect on pHi of hASMCs in static culture. 4-Acetamido-4'-isothio-cyanatostilbene-2,2'-disulfonic acid (SITS, 1 mM), a Cl-/HCO3- exchange inhibitor, affected the pHi of hASMCs both in static and flow conditions. Our results suggest that flow may perturb the Na+/H+ exchanger leading to an alkalinization of hASMCs, a different response from the flow-induced acidification seen with endothelial cells at the same levels of shear stress. Understanding the flow-induced signal transduction pathways in the vascular cells is of great importance in the tissue engineering of vascular grafts. In the case of SMCs, the involvement of pHi changes in nitric oxide production and proliferation regulation highlights further the significance of such studies.

NASA Discipline Cell Biology↗

High Ph, Ammonia Toxicity, and the Search for Life on the Jovian Planets

The effects of pH and ammonia concentration were studied separately, where possible, on a variety of organisms, including some isolated from natural environments of high pH and/or ammonia concentration. Escherichia coli and Bacillus subtilis are both extremely sensitive to ammonia. An aerobic organism (growth up to pH 11.4) from an alkaline spring is more resistant, but exhibits a toxic response to ammonia at a pH much lower than its maximum for growth. The greatest ammonia resistance has been found in an unidentified organism growing at near neutral pH. Even in this case, however, urvival at ammonia concentrations reasonably expected on the Jovian planets is measured in hours. This is two to three orders of magnitude longer than for E. coli. Results support the tentative conclusion that contamination of the Jovian planets with terrestrial organisms that can grow is unlikely. However, the range of toxic response noted, coupled with the observation that terrestrial life has not been exposed to high ammonia concentrations for millions of years, suggests that adaptation to greater ammonia tolerance may be possible.

Deal, P. H.↗

Properties of ribulose diphosphate carboxylase immobilized on porous glass

Ribulose-1,5-diphosphate carboxylase from spinach has been bound to arylamine porous glass with a diazo linkage and to alklamine porous glass with glutaraldehyde. Stability at elevated temperatures and responses to changes of pH and ribulose-1,5-diphosphate, Mg(2+), and dithiothreitol concentrations were not significantly different from the soluble enzyme, though stability at 4 C was somewhat improved.

Shapira, J.↗

Cellular mechanisms underlying growth asymmetry during stem gravitropism

Plant stems respond to gravitropic stimulation with a rapid, local and reversible change in cell growth rate (elongation), generally on both the upper and lower sides of the stem. The cellular and biochemical mechanisms for this differential growth are reviewed. Considerable evidence implicates an asymmetry in wall pH in the growth response. The strengths and weaknesses of the wall "loosening enzyme" concept are reviewed and the possibility of expansin involvement in the bending response of stems is considered. Also discussed is the possibility that wall stiffening processes, e.g. phenolic coupling driven by oxidative bursts or altered orientation of newly deposited cellulose, might mediate the growth responses during gravitropism.

NASA Discipline Plant Biology↗

Tomato responses to ammonium and nitrate nutrition under controlled root-zone pH

Tomato (Lycopersicon esculentum L. Mill. 'Vendor') plants were grown for 21 days in flowing solution culture with N supplied as either 1.0 mM NO3- or 1.0 mM NH4+. Acidity in the solutions was automatically maintained at pH 6.0. Accumulation and distribution of dry matter and total N and net photosynthetic rate were not affected by source of N. Thus, when rhizosphere acidity was controlled at pH 6.0 during uptake, either NO3- or NH4+ can be used efficiently by tomato. Uptake of K+ and Ca2+ were not altered by N source, but uptake of Mg2+ was reduced in NH4(+)-fed plants. This indicates that uptake of Mg2+ was regulated at least partially by ionic balance within the plant.

NASA Program CELSS↗

Thermally altered palagonitic tephra - A spectral and process analog to the soil and dust of Mars

Six palagonitic soil samples (PH-1 through PH-6) which were collected at 30-cm intervals from a lava slab on Mauna Kea, Hawaii, are studied. The samples present an alteration sequence caused by heating during emplacement of molten lava over a preexisting tephra cone. Techniques employed include visible and near-IUR spectroscopy, Moessbauer spectroscopy, and magnetic analysis. The four samples closest to the slab (PH-1 through PH-4) were strongly altered in response to heating during its emplacement; their iron oxide mineralogy is dominated by nanophase ferric oxide. The sample adjacent to the slab (PH-1) has a factor of 3 less H2O and contains crystalline hematite and magnetite in addition to nanophase ferric oxide. It is argued that localized thermal alteration events may provide a volumetrically important mechanism for the palagonitization of basaltic glass and the production of crystalline ferric oxides on Mars.

Bell, James F., III↗

Ecosystem and physiological controls over methane production in northern wetlands

Peat chemistry appears to exert primary control over methane production rates in the Canadian Northern Wetlands Study (NOWES) area. We determined laboratory methane production rate potentials in anaerobic slurries of samples collected from a transect of sites through the NOWES study area. We related methane production rates to indicators of resistance to microbial decay (peat C: N and lignin: N ratios) and experimentally manipulated substrate availability for methanogenesis using ethanol (EtOH) and plant litter. We also determined responses of methane production to pH and temperature. Methane production potentials declined along the gradient of sites from high rates in the coastal fens to low rates in the interior bogs and were generally highest in surface layers. Strong relationships between CH4 production potentials and peat chemistry suggested that methanogenesis was limited by fermentation rates. Methane production at ambient pH responded strongly to substrate additions in the circumneutral fens with narrow lignin: N and C: N ratios (delta CH4/delta EtOH = 0.9-2.3 mg/g) and weakly in the acidic bogs with wide C: N and lignin: N ratios (delta CH4/delta EtOH = -0.04-0.02 mg/g). Observed Q(sub 10) values ranged from 1.7 to 4.7 and generally increased with increasing substrate availability, suggesting that fermentation rates were limiting. Titration experiments generally demonstrated inhibition of methanogenesis by low pH. Our results suggest that the low rates of methane emission observed in interior bogs during NOWES likely resulted from pH and substrate quality limitation of the fermentation step in methane production and thus reflect intrinsically low methane production potentials. Low methane emission rates observed during NOWES will likely be observed in other northern wetland regions with similar vegetation chemistry.

Valentine, David W.↗

Paraventricular Stimulation with Glutamate Elicits Bradycardia and Pituitary Responses

The excitatory neurotransmitter, L-glutamate (0.5 M, pH 7.4), or the organic acid, acetate (0.5 M, pH 7.4), was microinjected (50 nl over 2 min) directly into the paraventricular nuclei (PVN) of pentobarbital sodium-anesthetized rats while arterial blood pressure and heart rate and plasma adrenocorticotropic hormone (ACTH), vasopressin, and oxytocin were measured. Activation of PVN neurons with L-glutamate led to increases in plasma ACTH, vasopressin, and oxytocin and a profound bradycardia (-80 beats/min) with little change in arterial blood pressure. Microinjection of acetate had no effect on the above variables. The decrease in heart rate was shown to be dependent on the concentration of glutamate injected and the volume of injectate. The bradycardia was mediated through the autonomic nervous system because ganglionic blockade (pentolinium tartrate) eliminated the response; atropine and propranolol severely attenuated the bradycardia. The bradycardia was greatest when L-glutamate was microinjected into the caudal PVN. Injections into the rostral PVN or into nuclei surrounding the PVN led to small or nonsignificant decreases in heart rate. Focal electric stimulation (2-50 pA) of the PVN also led to decreases in heart rate and arterial blood pressure. These data suggest that activation of PVN neurons leads to the release of ACTH, vasopressin, and oxytocin from the pituitary and a bradycardia that is mediated by the autonomic nervous system.

Darlington, Daniel N.↗

Temporal and vertical distribution of acidity and ionic composition in clouds - Comparison between modeling results and observations

Cyclic temporal variations of pH and ionic concentration in sampled clouds which traversed the Mt. Mitchell State Park (North Carolina) site during the summers of 1986, 1987, and 1988 are reported. These clouds typically had a measured pH minimum during their initial and final stages. The cause of this basic cylic pattern is attributed to sampling at different vertical levels of the cloud. This is substantiated by visual observations made while sampling. The results also suggest that the measured pH patterns do not always exhibit minima during the formative and dissipative stages of the cloud, apparently in response to the underlying dynamical processes. The relationship between temporal pH measurements made at a stationary site to vertical cloud levels provides insights into the physical processes (e.g., nucleation scavenging near cloud base, dry air entrainment near cloud top) influencing the observed cloud-water chemistry on a real-time basis and would improve cloud chemistry models.

Defelice, T. P.↗

Dynamics of vegetation and soils of oak/saw palmetto scrub after fire: Observations from permanent transects

Ten permanent 15 m transects previously established in two oak/saw palmetto scrub stands burned in December 1986, while two transects remained unburned. Vegetation in the greater than 0.5 m and the less than 0.5 m layers on these transects was sampled at 6, 12, 18, 24, and 36 months postburn and determined structural features of the vegetation (height, percent bare ground, total cover). The vegetation data were analyzed from each sampling by height layer using detrended correspondence analysis ordination. Vegetation data for the greater than 0.5 m layer for the entire time sequence were combined and analyzed using detrended correspondence analysis ordination. Soils were sampled at 6, 12, 18, and 24 months postburn and analyzed for pH, conductivity, organic matter, exchangeable cations (Ca, Mg, K, Na), NO3-N, NH4-N, Al, available metals (Cu, Fe, Mn, Zn), and PO4-P. Shrub species recovered at different rates postfire with saw palmetto reestablishing cover greater than 0.5 m within one year, but the scrub oaks had not returned to preburn cover greater than 0.5 m in 3 years after the fire. These differences in growth rates resulted in dominance shifts after the fire with saw palmetto increasing relative to the scrub oaks. Overall changes in species richness were minor, although changes occurred in species richness by height layers due to different growth rates. Soils of well drained and poorly drained sites differed markedly. Soil responses to the fire appeared minor. Soil pH increased at 6 and 12 months postfire; calcium increased at 6 months postburn. Nitrate-nitrogen increased at 12 months postburn. Low values of conductivity, PO4-P, Mg, K, Na, and Fe at 12 months postburn may be related to heavy rainfall the preceding month. Seasonal variability in some soil parameters appeared to occur.

Schmalzer, Paul A.↗

Ionic signaling in plant gravity and touch responses

Plant roots are optimized to exploit resources from the soil and as each root explores this environment it will encounter a range of biotic and abiotic stimuli to which it must respond. Therefore, each root must possess a sensory array capable of monitoring and integrating these diverse stimuli to direct the appropriate growth response. Touch and gravity represent two of the biophysical stimuli that plants must integrate. As sensing both of these signals requires mechano-transduction of biophysical forces to biochemical signaling events, it is likely that they share signal transduction elements. These common signaling components may allow for cross-talk and so integration of thigmotropic and gravitropic responses. Indeed, signal transduction events in both plant touch and gravity sensing are thought to include Ca(2+)- and pH-dependent events. Additionally, it seems clear that the systems responsible for root touch and gravity response interact to generate an integrated growth response. Thus, primary and lateral roots of Arabidopsis respond to mechanical stimuli by eliciting tropic growth that is likely part of a growth strategy employed by the root to circumvent obstacles in the soil. Also, the mechano-signaling induced by encountering an obstacle apparently down-regulates the graviperception machinery to allow this kind of avoidance response. The challenge for future research will be to define how the cellular signaling events in the root cap facilitate this signal integration and growth regulation. In addition, whether other stimuli are likewise integrated with the graviresponse via signal transduction system cross-talk is an important question that remains to be answered.

Review↗

In Vivo Model to Test Implanted Biosensors for Blood pH

Biosensors for monitoring physiologic data continuously through telemetry are available for heart rate, respiration, and temperature but not for blood pH or ions affected by hydrogen ion concentration. A telemetric biosensor for monitoring blood pH on-line could be used to identify and manage problems in fluid and electrolyte metabolism, cardiac and respiratory function during space flight and the acid-base status of patients without the need for venipuncture in patients on Earth. Critical to the development of biosensors is a method for evaluating their performance after implantation. Mature rats, prepared with jugular, cannulas for repeated blood samples, were exposed to a gas mixture containing high levels of carbon dioxide (7%) in a closed environment to induce mild respiratory acidosis. Serial blood gas and pH measurements in venous blood were compared with electrical responses from sensors implanted in the subcutaneous tissue. Animals became slightly tachypneic after exposure to excess CO2, but remained alert and active. After 5 minutes, basal blood pH decreased from 7.404 +/- 0.013 to 7.289 +/- 0.010 (p less than 0.001)and PC02 increased from 45 +/- 6 to 65 +/- 4 mm. Hg (p les than 0.001). Thereafter pH and blood gas parameters remained stable. Implanted sensors showed a decrease in millivolts (mV) which paralleled the change in pH and averaged 5-6 mV per 0.1 unit pH. Implanted sensors remained sensitive to modest changes in tissue pH for one week. A system for inducing acidosis in rats was developed to test the in vivo performance of pH biosensors. The system provides a method which is sensitive, rapid and reproducible in the same and different animals with full recovery, for testing the performance of sensors implanted in subcutaneous tissues.

Arnaud, Sara B.↗

Auxin-induced growth of Avena coleoptiles involves two mechanisms with different pH optima

Although rapid auxin-induced growth of coleoptile sections can persist for at least 18 hours, acid-induced growth lasts for a much shorter period of time. Three theories have been proposed to explain this difference in persistence. To distinguish between these theories, the pH dependence for auxin-induced growth of oat (Avena sativa L.) coleoptiles has been determined early and late in the elongation process. Coleoptile sections from which the outer epidermis was removed to facilitate buffer entry were incubated, with or without 10 micromolar indoleacetic acid, in 20 millimolar buffers at pH 4.5 to 7.0 to maintain a fixed wall pH. During the first 1 to 2 hours after addition of auxin, elongation occurs by acid-induced extension (i.e. the pH optimum is <5 and the elongation varies inversely with the solution pH). Auxin causes no additional elongation because the buffers prevent further changes in wall pH. After 60 to 90 minutes, a second mechanism of auxin-induced growth, whose pH optimum is 5.5 to 6.0, predominates. It is proposed that rapid growth responses to changes in auxin concentration are mediated by auxin-induced changes in wall pH, whereas the prolonged, steady-state growth rate is controlled by a second, auxin-mediated process whose pH optimum is less acidic.

NASA Program Space Biology↗

Evaluation of a Multi-Parameter Sensor for Automated, Continuous Cell Culture Monitoring in Bioreactors

Compact and automated sensors are desired for assessing the health of cell cultures in biotechnology experiments in microgravity. Measurement of cell culture medium allows for the optirn.jzation of culture conditions on orbit to maximize cell growth and minimize unnecessary exchange of medium. While several discrete sensors exist to measure culture health, a multi-parameter sensor would simplify the experimental apparatus. One such sensor, the Paratrend 7, consists of three optical fibers for measuring pH, dissolved oxygen (p02), dissolved carbon dioxide (pC02) , and a thermocouple to measure temperature. The sensor bundle was designed for intra-arterial placement in clinical patients, and potentially can be used in NASA's Space Shuttle and International Space Station biotechnology program bioreactors. Methods: A Paratrend 7 sensor was placed at the outlet of a rotating-wall perfused vessel bioreactor system inoculated with BHK-21 (baby hamster kidney) cells. Cell culture medium (GTSF-2, composed of 40% minimum essential medium, 60% L-15 Leibovitz medium) was manually measured using a bench top blood gas analyzer (BGA, Ciba-Corning). Results: A Paratrend 7 sensor was used over a long-term (>120 day) cell culture experiment. The sensor was able to track changes in cell medium pH, p02, and pC02 due to the consumption of nutrients by the BHK-21. When compared to manually obtained BGA measurements, the sensor had good agreement for pH, p02, and pC02 with bias [and precision] of 0.02 [0.15], 1 mm Hg [18 mm Hg], and -4.0 mm Hg [8.0 mm Hg] respectively. The Paratrend oxygen sensor was recalibrated (offset) periodically due to drift. The bias for the raw (no offset or recalibration) oxygen measurements was 42 mm Hg [38 mm Hg]. The measured response (rise) time of the sensor was 20 +/- 4s for pH, 81 +/- 53s for pC02, 51 +/- 20s for p02. For long-term cell culture measurements, these response times are more than adequate. Based on these findings , the Paratrend sensor could offer automated, continuous monitoring of cell cultures with a temporal resolution of 1 minute, which is not attainable by sampling via handheld blood analyzer (i-STAT). Conclusion: The resulting bias and precision found in these cell culture-based studies is comparable to Paratrend sensor clinical results. Although the large error in p02 measurements (+/-18 mm Hg) may be acceptable for clinical applications, where Paratrend values are periodically adjusted to a BGA measurement, the O2 sensor in this bundle may not be reliable enough for the single-calibration requirement of sensors used in NASA's bioreactors. The pH and pC02 sensors in the bundle are reliable and stable over the measurement period, and can be used without recalibration to measure cell cultures in rn.jcrogravity biotechnology experiments. Future work will test additional Paratrend sensors to provide statistical assessment of sensor performance.

Pappas, D.↗

Generation of avian cells resembling osteoclasts from mononuclear phagocytes

Several lines of indirect evidence suggest that a monocyte family precursor gives rise to the osteoclast, although this hypothesis is controversial. Starting with a uniform population of nonspecific esterase positive, tartrate-sensitive, acid phosphatase-producing, mannose receptor-bearing mononuclear cells, prepared from dispersed marrow of calcium-deprived laying hens by cell density separation and selective cellular adherence, we generated multinucleated cells in vitro. When cultured with devitalized bone, these cells show, by electron microscopy, the characteristic osteoclast morphology in that they are mitochondria-rich, multinucleated, and, most importantly, develop characteristic ruffled membranes at the matrix attachment site. Moreover, as documented by scanning electron microscopy, these cells pit bone slices in a manner identical to freshly isolated osteoclasts. In addition, isoenzymes of acid phosphatase from generated osteoclasts, separated by 7.5% polyacrylamide gel electrophoresis at pH 4, are identical to those of mature osteoclasts in migration pattern and tartrate resistance, although the precursor cells from which the osteoclasts are generated produce an entirely different isoenzyme, which is tartrate-sensitive and migrates less rapidly at pH 4. The fused cells also exhibit a cAMP response to prostaglandin E2. Therefore, osteoclast-like cells can be derived by in vitro culture of a marrow-derived monocyte cell population.

NASA Discipline Musculoskeletal↗

Dimensional stability considerations for cryogenic metals

Work performed as part of an effort to identify, and where possible separate out, some of the factors that contribute to dimensional stability in cryogenic wind tunnel models is reported. Initial problems were encountered with two dimensional models made of 15-5 PH stainless steel, which warped significantly after being subjected to cryogenic testing in the 0.3 Meter Transonic Cryogenic Tunnel. Subsequently, an effort was undertaken to investigate the mechanisms that could cause model warpage during cryogenic testing. The two basic mechanisms that can lead to warpage are (1) metallurgical structural instability in which one phase transforms partially or fully into a second phase which has a different crystal structure and volume, and (2) deformation due to the creation, or relief, of unbalanced induced or residual stresses. In the case of the 15-5 PH airfoils, it is highly probable that metallurgical instability was responsible for most of the observed warpage. A particular specimen configuration was established for use in the systematic evaluation of the factors influencing warpage. Preliminary studies of a specimen made of VASCOMAX 200 suggest the possibility of manipulating the stresses in the surface layers by appropriate combinations of milling and grinding steps. This opens up the possibility of correcting or establishing the required surface profile of an airfoil.

Wigley, D. A.↗

Chemochromic Indicators for the Detection of Hypergolic Fuels

The toxicity and hazard level associated with the use of hypergolic fuels necessitates the development of technology capable of detecting the presence of such fuels in a variety of different environments and conditions. The most commonly used sensors for the detection of hypergolic fuels are electrochemical in nature, which have serious limitations when used as area monitoring devices. Recent collaborative work between Kennedy Space Center and ASRC Aerospace has led to the development of indicators which exhibit a color change upon exposure to hydrazine under different conditions. The indicators under investigation on this developmental effort are para-dimethylaminobenzaldehyde (PDAB), various formulations of universal pH indicators, and potassium tetrachloroaurate (KAuCl4). These chemochromic indicators have been tested for the detection of hydrazine under various conditions: pure liquid fuel, aqueous fuel solution, saline aqueous fuel solutions, vapor fuel, and 3-month shelf life study, which included UV protection, thermal extremes, and normal storage conditions. The hypergolic fuel indicator test was conducted with the indicator impregnated into a wipe material to test the applicability of the indicator to be used to capture (absorb) and indicate the presence of hypergolic fuels. Each of the indicators performed well, with the universal pH indicator being the best candidate because of the visible response color change and the indicator stability after the shelf life study.

Santiago-Maldonado, E.↗