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Sample records for activity aw ph

  1. Effects of pH or a(w) stress on growth of Listeria monocytogenes.

    PubMed

    Cheroutre-Vialette, M; Lebert, I; Hebraud, M; Labadie, J C; Lebert, A

    1998-06-30

    The growth of three strains of Listeria monocytogenes at 20 degrees C in a meat broth of different pH or water activity was investigated. At inoculation or at the beginning of the exponential phase, cells were exposed to stress by the addition of NaOH or NH4+, acetic acid, NaCl or KCl, in order to reach a pH of either 9.0 or 5.6, or an a(w) of 0.950 or 0.965, respectively. The effects of the exposure to stress on the generation and lag times of each strain were analysed by turbidity measurements for cultures in micro-titer plates. Results were confirmed by conducting the same experiments in a fermentor, except for the maximal population reached. The three strains showed similar behaviour. Cells were able to overcome the alkaline stress rapidly whereas acid and osmotic shocks induced important changes of the growth parameters. Cells exposed to acid or osmotic conditions from the time of inoculation were less affected than cells exposed at the beginning of the mid-exponential phase.

  2. Impact of different water activities (aw) adjusted by solutes on high pressure high temperature inactivation of Bacillus amyloliquefaciens spores

    PubMed Central

    Sevenich, Robert; Reineke, Kai; Hecht, Philipp; Fröhling, Antje; Rauh, Cornelia; Schlüter, Oliver; Knorr, Dietrich

    2015-01-01

    Much research has been conducted to comprehend the mechanisms of high pressure (HP) inactivation of spores in aqueous systems but for food model systems these information are scarce. In these systems spores can interact with ingredients which then could possibly lead to retarded or reduced inactivation, which can cause a problem for the sterilization process. The protective mechanism of a reduced aw-value is still unclear. HP processing might prove valuable to overcome protective effects of solutes and achieve shorter process times for sterilization under HP. To gain insight into the underlying mechanisms five aw-values (0.9, 0.92, 0.94, 0.96, 1) were adjusted with two different solutes (NaCl, sucrose). Solutions were inoculated with spores of Bacillus amyloliquefaciens and treated at 105, 110, and 115°C at 600 MPa. Further a thermal inactivation was conducted at the same temperatures for a comparison with the HP data. Afterward, the influence of HP high temperature treatment on the inactivation, the dipicolinic acid (DPA)-release and membrane constitution was assessed by plate count, HPLC and flow cytometry (FCM). The results show that during HP treatments sucrose and salt both have a protective effect, in which the influence of sucrose on the retarded inactivation is higher. The threshold water activities (aw), which is 0.94, here salt and sucrose have a significant influence on the inactivation. The comparison of thermal (105–115°C) and HP and high temperature (600 MPa, 105–115°C) treated samples showed that the time needed to achieve a 4–5 log10 inactivation is reduced from 45 (aw = 1) to 75 (aw = 0.9) min at 105°C to 3 (aw = 1) to 15 (aw = 0.9) minutes at 600 MPa and 105°C. The release of DPA is the rate limiting step of the inactivation and therefore monitoring the release is of great interest. The DPA-release is slowed down in high concentrated solutions (e.g., sucrose, salt) in comparison to aw 1. Since there is a difference in the way the

  3. Effects of dehydroepiandrosterone on obesity and glucose-6-phosphate dehydrogenase activity in the lethal yellow mouse (strain 129/Sv-Ay/Aw).

    PubMed

    Granholm, N H; Staber, L D; Wilkin, P J

    1987-04-01

    We investigated the anti-obesity effects of the adrenal androgen, dehydroepiandrosterone (DHEA), on genetically predisposed obese lethal yellow mice (Ay/Aw). Secondly, we tested the hypothesis that DHEA promotes its anti-obesity effects by decreasing the activity of glucose-6-phosphate dehydrogenase (G6PDH). We subjected four genotype-sex combinations of yellow and agouti (control) mice to four dietary treatments and determined weight changes, food consumption, and G6PDH activity. Although G6PDH activities of yellow mice were considerably decreased in the 0.4% DHEA treatment group, they were elevated in the 0.0 and 0.1% DHEA treatment groups. In contrast, G6PDH activities of DHEA-treated control agouti mice remained relatively constant. These studies confirm that DHEA prevents the Ay gene from promoting excess fat deposition via some mechanism(s) other than reduced dietary intake. However, the overall absence of agreement between weight change (gain or loss) and G6PDH activity suggests that the anti-obesity activity of DHEA is not mediated via G6PDH. Since yellow obese (Ay/Aw) mice were found to be more susceptible to DHEA's effects than their agouti (Aw/Aw) littermates, Ay appears to induce an altered metabolism in Ay/Aw mice which is more susceptible to the effects of DHEA than the normal metabolism of Aw/Aw mice.

  4. Estimation of water activity from pH and °Brix values of some food products.

    PubMed

    Gabriel, Alonzo A

    2008-06-01

    In this study, a predictive model for the estimation of water activity (aw(25°C)) as a function of pH (1.00-8.00) and °Brix (0-82.00) values of simulated food solutions (SFS) was developed, through response surface methodology. Response fit analyses resulted in a highly significant (pH<0.0001) square root polynomial model that can predict aw(25°C) of SFS in terms of pH and °Brix values within the defined variable ranges. The linear, quadratic and interactive influences of pH and °Brix on aw(25°C) were all significant (pH<0.0001). Model validations in SFS and in a number of actual food systems showed that the model had acceptable predictive performance, as indicated by the calculated accuracy and bias indices.

  5. Modelling the unexpected effect of acetic and lactic acid in combination with pH and aw on the growth/no growth interface of Zygosaccharomyces bailii.

    PubMed

    Vermeulen, A; Dang, T D T; Geeraerd, A H; Bernaerts, K; Debevere, J; Van Impe, J; Devlieghere, F

    2008-05-10

    Microbial spoilage of shelf-stable acidified sauces is predominantly caused by lactic acid bacteria and yeasts. A specific spoilage yeast in these products is Zygosaccharomyces bailii, as this fructophilic, osmotolerant, and weak acid resistant yeast is difficult to control. A growth/no growth model was developed describing the influence of (i) pH in a range from pH 3.0 to pH 5.0 (5 levels), (ii) acetic acid in a range from 0 to 3.5% (w/v), and (iii) lactic acid in a range from 0 to 3.0% (w/v). aw was fixed at a level of 0.95 which is representative for acidified sauces with high sugar content. Modified Sabouraud medium was inoculated at +/- 10(4) CFU/ml, incubated at 30 degrees C and growth was assessed by optical density measurements. All combinations of environmental conditions were tested in at least twelve replicates, yielding precise values for the probability of growth. Results showed that replacing acetic acid by lactic acid, which has a milder taste, may imply some risks on food spoilage because, under some conditions, stimulation of growth by lactic acid was observed. This stimulation had also consequences on the model development: (i) only ordinary logistic regression models were able to describe this phenomenon due to their flexible behaviour, (ii) it was necessary to split up the data set into two subsets to have the best description of the obtained data. Two different ordinary logistic regression models were fitted on these data sets taking either the total acid concentration as one of the explanatory variables or differentiating between the undissociated and dissociated acid concentrations. The obtained models were compared with the CIMSCEE code [CIMSCEE, 1992. Code for the production of microbiologically safe and stable emulsified and non-emulsified sauces containing acetic acid. Comité des Industries des Mayonnaise et Sauces Condimentaires, de la Communauté Economique Européenne, Brussels, Belgium], a formula which is nowadays often used by the

  6. Probabilistic modeling of Saccharomyces cerevisiae inhibition under the effects of water activity, pH, and potassium sorbate concentration.

    PubMed

    López-Malo, A; Guerrero, S; Alzamora, S M

    2000-01-01

    Probabilistic microbial modeling using logistic regression was used to predict the boundary between growth and no growth of Saccharomyces cerevisiae at selected incubation periods (50 and 350 h) in the presence of growth-controlling factors such as water activity (a(w); 0.97, 0.95, and 0.93), pH (6.0, 5.0, 4.0, and 3.0), and potassium sorbate (0, 50, 100, 200, 500, and 1,000 ppm). The proposed model predicts the probability of growth under a set of conditions and calculates critical values of a(w), pH, and potassium sorbate concentration needed to inhibit yeast growth for different probabilities. The reduction of pH increased the number of combinations of a(w) and potassium sorbate concentration with probabilities to inhibit yeast growth higher than 0.95. With a probability of growth of 0.05 and using the logistic models, the critical pH values were higher for 50 h of incubation than those required for 350 h. With lower a(w) values and increasing potassium sorbate concentration the critical pH values increased. Logistic regression is a useful tool to evaluate the effects of the combined factors on microbial growth.

  7. Decision Document for the Low Activity Waste Retrieval Strategy for Tanks 241-AN-103 and 241-AN-104 and 241-AN-105 and 241-AW-101

    SciTech Connect

    RASMUSSEN, O.R.

    2000-09-28

    This report documents the preferred approach (retrieval strategy) to prepare and transfer waste from low-activity waste source tanks containing soluble solids (Tanks 241-AN-103, 241-AN-104, 241-AN-105 and 241-AW-101) to the vitrification plant. Several opportunities to further refine the selected retrieval strategy were identified; these were recommended for follow-on studies.

  8. Temperature, water activity and pH during conidia production affect the physiological state and germination time of Penicillium species.

    PubMed

    Nguyen Van Long, Nicolas; Vasseur, Valérie; Coroller, Louis; Dantigny, Philippe; Le Panse, Sophie; Weill, Amélie; Mounier, Jérôme; Rigalma, Karim

    2017-01-16

    Conidial germination and mycelial growth are generally studied with conidia produced under optimal conditions to increase conidial yield. Nonetheless, the physiological state of such conidia most likely differs from those involved in spoilage of naturally contaminated food. The present study aimed at investigating the impact of temperature, pH and water activity (aw) during production of conidia on the germination parameters and compatible solutes of conidia of Penicillium roqueforti and Penicillium expansum. Low temperature (5°C) and reduced aw (0.900 aw) during sporulation significantly reduced conidial germination times whereas the pH of the sporulation medium only had a slight effect at the tested values (2.5, 8.0). Conidia of P. roqueforti produced at 5°C germinated up to 45h earlier than those produced at 20°C. Conidia of P. roqueforti and P. expansum produced at 0.900 aw germinated respectively up to 8h and 3h earlier than conidia produced at 0.980 aw. Furthermore, trehalose and mannitol assessments suggested that earlier germination might be related to delayed conidial maturation even though no ultra-structural modifications were observed by transmission electron microscopy. Taken together, these results highlight the importance of considering environmental conditions during sporulation in mycological studies. The physiological state of fungal conidia should be taken into account to design challenge tests or predictive mycology studies. This knowledge may also be of interest to improve the germination capacity of fungal cultures commonly used in fermented foods.

  9. Modelling biofilm formation of Salmonella enterica ser. Newport as a function of pH and water activity.

    PubMed

    Dimakopoulou-Papazoglou, Dafni; Lianou, Alexandra; Koutsoumanis, Konstantinos P

    2016-02-01

    The effect of pH and water activity (aw) on the formation of biofilm by Salmonella enterica ser. Newport, previously identified as a strong biofilm producer, was assessed. Biofilm formation was evaluated in tryptone soy broth at 37 °C and at different combinations of pH (3.3-7.8) and aw (0.894-0.997). In total, 540 biofilm formation tests in 108 pH and aw combinations were carried out in polystyrene microtiter plates using crystal violet staining and optical density (OD; 580 nm) measurements. Since the individual effects of pH and aw on biofilm formation had a similar pattern to that observed for microbial growth rate, cardinal parameter models (CPMs) were used to describe these effects. CPMs described successfully the effects of these two environmental parameters, with the estimated cardinal values of pHmin, pHopt, pHmax, awmin and awopt being 3.58, 6.02, 9.71, 0.894 and 0.994, respectively. The CPMs assumption of the multiplicative inhibitory effect of environmental factors was validated in the case of biofilm formation using additional independent data (i.e. 430 OD data at 86 different combinations of pH and aw). The validation results showed a good agreement (r(2) = 0.938) between observed and predicted OD with no systematic error. In the second part of this study, a probabilistic model predicting the pathogen's biofilm formation boundaries was developed, and the degree of agreement between predicted probabilities and observations was as high as 99.8%. Hence, the effect of environmental parameters on biofilm formation can be quantitatively expressed using mathematical models, with the latter models, in turn, providing useful information for biofilm control in food industry environments.

  10. Modified atmosphere packaging for prevention of mold spoilage of bakery products with different pH and water activity levels.

    PubMed

    Guynot, M E; Marín, S; Sanchis, V; Ramos, A J

    2003-10-01

    A sponge cake analog was used to study the influence of pH, water activity (aw), and carbon dioxide (CO2) levels on the growth of seven fungal species commonly causing bakery product spoilage (Eurotium amstelodami, Eurotium herbariorum, Eurotium repens, Eurotium rubrum, Aspergillus niger, Aspergillus flavus, and Penicillium corylophilum). A full factorial design was used. Water activity, CO2, and their interaction were the main factors significantly affecting fungal growth. Water activity at levels of 0.80 to 0.90 had a significant influence on fungal growth and determined the concentration of CO2 needed to prevent cake analog spoilage. At an aw level of 0.85, lag phases increased twofold when the level of CO2 in the headspace increased from 0 to 70%. In general, no fungal growth was observed for up to 28 days of incubation at 25 degrees C when samples were packaged with 100% CO2, regardless of the aw level. Partial least squares projection to latent structures regression was used to build a polynomial model to predict sponge cake shelf life on the basis of the lag phases of all seven species tested. The model developed explained quite well (R2 = 79%) the growth of almost all species, which responded similarly to changes in tested factors. The results of this study emphasize the importance of combining several hurdles, such as modified atmosphere packaging, aw, and pH, that have synergistic or additive effects on the inhibition of mold growth.

  11. Preclinical immunogenicity and functional activity studies of an A+W meningococcal outer membrane vesicle (OMV) vaccine and comparisons with existing meningococcal conjugate- and polysaccharide vaccines.

    PubMed

    Tunheim, G; Arnemo, M; Næss, L M; Fjeldheim, Å K; Nome, L; Bolstad, K; Aase, A; Mandiarote, A; González, H; González, D; García, L; Cardoso, D; Norheim, G; Rosenqvist, E

    2013-12-09

    Meningococci of serogroups A and W (MenA and MenW) are the main causes of epidemic bacterial meningitis outbreaks in sub-Saharan Africa. In this study we prepared a detergent extracted outer membrane vesicle (dOMV) vaccine from representative African MenA and MenW strains, and compared the immunogenicity of this vaccine with existing meningococcal conjugate and polysaccharide (PS) vaccines in mice. NMRI mice were immunized with preclinical batches of the A+W dOMV vaccine, or with commercially available vaccines; a MenA conjugate vaccine (MenAfriVac(®), Serum Institute of India), ACYW conjugate vaccine (Menveo(®), Novartis) or ACYW PS vaccine (Mencevax(®), GlaxoSmithKline). The mice received 2 doses of 1/10 or 1/50 of a human dose with a three week interval. Immune responses were tested in ELISA, serum bactericidal activity (SBA) and opsonophagocytic activity (OPA) assays. High levels of IgG antibodies against both A and W dOMV were detected in mice receiving the A+W dOMV vaccine. High SBA titers against both MenA and MenW vaccine strains were detected after only one dose of the A+W dOMV vaccine, and the titers were further increased after the second dose. The SBA and OPA titers in mice immunized with dOMV vaccine were significantly higher than in mice immunized with the ACYW-conjugate vaccine or the PS vaccine. Furthermore, the A+W dOMV vaccine was shown to induce SBA and OPA titers against MenA of the same magnitude as the titers induced by the A-conjugate vaccine. In conclusion, the A+W dOMV vaccine induced high levels of functional antibodies to both MenA and MenW strains, levels that were shown to be higher or equal to the levels induced by licensed meningococcal vaccines. Thus, an A+W dOMV vaccine could potentially serve as an alternative or a supplement to existing conjugate and PS vaccines in the African meningitis belt.

  12. Modeling the growth and death kinetics of Salmonella in poultry litter as a function of pH and water activity.

    PubMed

    Payne, J B; Osborne, J A; Jenkins, P K; Sheldon, B W

    2007-01-01

    Contaminated poultry litter, serving as a reservoir for Salmonella, can be linked to both food safety concerns when contaminated birds enter processing plants and environmental concerns when used as a fertilizer. Predictive modeling allows for the estimation of microbial growth or inactivation as a function of controlling environmental growth factors. A study was conducted to observe the combined effects of pH and water activity (A(w)) at a constant temperature on Salmonella populations in used turkey litter to predict microbial response over time. Litter, first pH-adjusted and then inoculated with a 3-strain Salmonella serovar cocktail to an initial concentration of approximately 10(7) cfu/g, was placed into individual sealed plastic containers with saturated salt solutions for controlling A(w). A balanced design including 3 A(w) values (0.84, 0.91, 0.96), 3 pH values (4, 7, 9), and a constant temperature of 30 degrees C was used, with litter samples periodically removed and analyzed for Salmonella populations, pH, and A(w). At each combination of environmental factors, the Churchill or exponential inactivation mathematical models were used to describe the growth and death of Salmonella over time. Salmonella populations exhibited growth (approximately 2 log) with little decline up to 42 d in litter environments of pH 7 and 9 and a A(w) of 0.96. As litter A(w) and pH levels were reduced, populations declined, with the most drastic reductions (approximately 5 log in 9 h) occurring in low-pH (4) and low-A(w) (0.84) environments. Generalized models for bacterial growth and death under grouped pH environments were successfully developed to predict Salmonella behavior in litter over time. These findings suggest that the best management practices and litter treatments that lower litter A(w) to < or =0.84 and pH to < or =4 are effective in reducing Salmonella populations. The use of a single equation to predict the growth and decline of Salmonella populations as a

  13. Effect of weak acid preservatives on growth of bakery product spoilage fungi at different water activities and pH values.

    PubMed

    Suhr, K I; Nielsen, P V

    2004-08-15

    Inhibition of spoilage organisms from bakery products by weak acid preservatives in concentrations of 0%, 0.003%, 0.03% and 0.3% (w/v) was investigated experimentally on a substrate media with water activity (a(w)) and pH ranging from sourdough-fermented acidic rye bread to alkaline intermediate moisture sponge cake types (a(w) 0.80-0.95, pH 4.7-7.4). Initially, rye bread conditions (a(w) 0.94-0.97 and pH 4.4-4.8) in combination with calcium propionate were investigated. Results showed that the highest concentration of propionate (0.3%) at all conditions apart from high a(w) (0.97) and high pH (4.8) totally inhibited fungal growth for a 2-week period, with the exception of Penicillium roqueforti, Penicillium commune and Eurotium rubrum. Characteristically for the major spoiler of rye bread, P. roqueforti, all three isolates tested were stimulated by propionate and the stimulation was significantly enhanced at high water activity levels. The effect of propionate on production of secondary metabolites (mycophenolic acid, rugulovasine, echinulin, flavoglaucin) was also studied, and variable or isolate dependent results were found. Subsequently, a screening experiment representing a wider range of bakery products was conducted using calcium propionate, potassium sorbate and sodium benzoate. The obtained data was modelled using survival analysis to determine 'spoilage-free time' for the fungi. At the low a(w) level (0.80) only Eurotium species grew within the test period of 30 days. Higher water activity levels as well as higher pH values decreased spoilage-free times of the fungi. The preservative calcium propionate was less effective than potassium sorbate and sodium benzoate.

  14. Effect of temperature, water activity, and pH on growth and production of ochratoxin A by Aspergillus niger and Aspergillus carbonarius from Brazilian grapes.

    PubMed

    Passamani, Fabiana Reinis Franca; Hernandes, Thais; Lopes, Noelly Alves; Bastos, Sabrina Carvalho; Santiago, Wilder Douglas; Cardoso, Maria das Graças; Batista, Luís Roberto

    2014-11-01

    The growth of ochratoxigenic fungus and the presence of ochratoxin A (OTA) in grapes and their derivatives can be caused by a wide range of physical, chemical, and biological factors. The determination of interactions between these factors and fungal species from different climatic regions is important in designing models for minimizing the risk of OTA in wine and grape juice. This study evaluated the influence of temperature, water activity (aw), and pH on the development and production of OTA in a semisynthetic grape culture medium by Aspergillus carbonarius and Aspergillus niger strains. To analyze the growth conditions and production of OTA, an experimental design was conducted using response surface methodology as a tool to assess the effects of these abiotic variables on fungal behavior. A. carbonarius showed the highest growth at temperatures from 20 to 33°C, aw between 0.95 and 0.98, and pH levels between 5 and 6.5. Similarly, for A. niger, temperatures between 24 and 37°C, aw greater than 0.95, and pH levels between 4 and 6.5 were optimal. The greatest toxin concentrations for A. carbonarius and A. niger (10 μg/g and 7.0 μg/g, respectively) were found at 15°C, aw 0.99, and pH 5.35. The lowest pH was found to contribute to greater OTA production. These results show that the evaluated fungi are able to grow and produce OTA in a wide range of temperature, aw, and pH. However, the optimal conditions for toxin production are generally different from those optimal for fungal growth. The knowledge of optimal conditions for fungal growth and production of OTA, and of the stages of cultivation in which these conditions are optimal, allows a more precise assessment of the potential risk to health from consumption of products derived from grapes.

  15. AW-101 entrained solids - Solubility versus temperature

    SciTech Connect

    GJ Lumetta; RC Lettau; GF Piepel

    2000-03-31

    This report describes the results of a test conducted by Battelle to assess the solubility of the solids entrained in the diluted AW-101 low-activity waste (LAW) sample. BNFL requested Battelle to dilute the AW-1-1 sample using de-ionized water to mimic expected plant operating conditions. BNFL further requested Battelle to assess the solubility of the solids present in the diluted AW-101 sample versus temperature conditions of 30, 40, and 50 C. BNFL requested these tests to assess the composition of the LAW supernatant and solids versus expected plant-operating conditions. The work was conducted according to test plan BNFL-TP-29953-7, Rev. 0, Determination of the Solubility of LAW Entrained Solids. The test went according to plan, with no deviations from the test plan.

  16. Quantification of the relative effects of temperature, pH, and water activity on inactivation of Escherichia coli in fermented meat by meta-analysis.

    PubMed

    McQuestin, Olivia J; Shadbolt, Craig T; Ross, Tom

    2009-11-01

    Outbreaks of Escherichia coli infections linked to fermented meats have prompted much research into the kinetics of E. coli inactivation during fermented meat manufacture. A meta-analysis of data from 44 independent studies was undertaken that allowed the relative influences of pH, water activity (a(w)), and temperature on E. coli survival during fermented meat processing to be investigated. Data were reevaluated to determine rates of inactivation, providing 484 rate data points with various pH (2.8 to 6.14), a(w) (0.75 to 0.986), and temperature (-20 to 66 degrees C) values, product formulations, and E. coli strains and serotypes. When the data were presented as an Arrhenius model, temperature (0 to 47 degrees C) accounted for 61% of the variance in the ln(inactivation rate) data. In contrast, the pH or a(w) measured accounted for less than 8% of variability in the data, and the effects of other pH- and a(w)-based variables (i.e., total decrease and rates of reduction of those factors) were largely dependent on the temperature of the process. These findings indicate that although temperatures typically used in fermented meat manufacture are not lethal to E. coli per se, when other factors prevent E. coli growth (e.g., low pH and a(w)), the rate of inactivation of E. coli is dominated by temperature. In contrast, inactivation rates at temperatures above approximately 50 degrees C were characterized by smaller z values than those at 0 to 47 degrees C, suggesting that the mechanisms of inactivation are different in these temperature ranges. The Arrhenius model developed can be used to improve product safety by quantifying the effects of changes in temperature and/or time on E. coli inactivation during fermented meat manufacture.

  17. Interactive effects of temperature, pH, and water activity on the growth kinetics of Shiga toxin-producing Escherichia coli O104:H4 3.

    PubMed

    Juneja, Vijay K; Mukhopadhyay, Sudarsan; Ukuku, Dike; Hwang, Cheng-An; Wu, Vivian C H; Thippareddi, Harshavardhan

    2014-05-01

    The risk of non-O157 Shiga toxin-producing Escherichia coli strains has become a growing public health concern. Several studies characterized the behavior of E. coli O157:H7; however, no reports on the influence of multiple factors on E. coli O104:H4 are available. This study examined the effects and interactions of temperature (7 to 46°C), pH (4.5 to 8.5), and water activity (aw ; 0.95 to 0.99) on the growth kinetics of E. coli O104:H4 and developed predictive models to estimate its growth potential in foods. Growth kinetics studies for each of the 23 variable combinations from a central composite design were performed. Growth data were used to obtain the lag phase duration (LPD), exponential growth rate, generation time, and maximum population density (MPD). These growth parameters as a function of temperature, pH, and aw as controlling factors were analyzed to generate second-order response surface models. The results indicate that the observed MPD was dependent on the pH, aw, and temperature of the growth medium. Increasing temperature resulted in a concomitant decrease in LPD. Regression analysis suggests that temperature, pH, and aw significantly affect the LPD, exponential growth rate, generation time, and MPD of E. coli O104:H4. A comparison between the observed values and those of E. coli O157:H7 predictions obtained by using the U. S. Department of Agriculture Pathogen Modeling Program indicated that E. coli O104:H4 grows faster than E. coli O157:H7. The developed models were validated with alfalfa and broccoli sprouts. These models will provide risk assessors and food safety managers a rapid means of estimating the likelihood that the pathogen, if present, would grow in response to the interaction of the three variables assessed.

  18. Estimating the non-thermal inactivation of Listeria monocytogenes in fermented sausages relative to temperature, pH and water activity.

    PubMed

    Mataragas, M; Rantsioua, K; Alessandria, V; Cocolin, L

    2015-02-01

    Data relative to in situ Listeria monocytogenes inactivation in fermented sausages were collected from 13 individual studies found in the literature. Inactivation rates were extrapolated and used to develop a predictive model to evaluate the relative effects of pH, water activity (a(w)) and temperature on L. monocytogenes fate during fermentation and ripening. Temperature explained ca. 60% of the data variability, while pH and a(w) only a small part. Temperature alone may not be sufficient to cause pathogen's inactivation, but inactivation rate is dominated by temperature when pH and a(w) are in the range which prevent L. monocytogenes growth. A predictive model based on two Arrhenius equations (ln[inactivation rate]=-25.71-[-0.6829 / (8.314 × T)] for fermentation; and ln[inactivation rate]=-44.86-[-1.219/(8.314 × T)] for ripening)was developed. The model can be used to quantify the effect of temperature and/or time changes on fermented sausage safety. The advantages and limitations of the model are discussed.

  19. Effects of a(w) packaging time and atmosphere composition on aroma profile, biogenic amine content and microbiological features of dry fermented sausages.

    PubMed

    Tabanelli, Giulia; Montanari, Chiara; Grazia, Luigi; Lanciotti, Rosalba; Gardini, Fausto

    2013-06-01

    The aim was to evaluate the effects of different water contents (corresponding to a(w) 0.940, 0.914, 0.895 and 0.875) of sausages at different times on the characteristics of typical Italian dry fermented sausages packaged under two different modified atmospheres (MAP) (100%N2 or 30%CO2/70%N2). During ripening and storage, samples were analyzed to evaluate pH and aw changes, microbial population, biogenic amine (BA) accumulation and aroma profile. The results showed that during storage under MAP several microbiological and enzymatic activities proceeded, leading to changes in aroma profile. In particular, higher hexanal concentrations in samples at lower aw and significant amounts of acetone, 2-butanone, ethanol and 2-butanol in samples at aw>0.900 were detected. Moreover, data showed modifications in BA content, such as higher accumulation in samples at aw>0.900 in MAP with 100%N2. This behavior depends, in first instance, on the aw of the sausages when packaged.

  20. Development of a predictive model for Salmonella spp. reduction in meat jerky product with temperature, potassium sorbate, pH, and water activity as controlling factors.

    PubMed

    Juneja, Vijay K; Valenzuela-Melendres, Martin; Heperkan, Dilek; Bautista, Derrick; Anderson, David; Hwang, Cheng-An; Peña-Ramos, Aida; Camou, Juan Pedro; Torrentera-Olivera, Noemi

    2016-11-07

    The objective of this study was to develop a predictive model for the inactivation of Salmonella spp. in ground beef jerky as a function of temperature (T), pH, potassium sorbate (PS), and final water activity (aw). Following a central composite design, ground beef was combined with PS (0 to 0.3%, w/w), pH adjusted from 5 to 7, inoculated with a cocktail of 6 serotypes of Salmonella spp. and heat processed at temperatures between 65 and 85°C until the final aw ranging from 0.65 to 0.85 was achieved. Surviving Salmonella cells were enumerated on tryptic soy agar overlaid with xylose lysine deoxycholate agar (pre-tempered to 47°C) after incubation for 48h at 30°C. Bacterial inactivation was quantified in terms of logarithmic reductions of Salmonella counts (log10CFU/g) and inactivation rate (log10(CFU/g)/h). The results indicated that pH, PS and T significantly (p<0.05) interacted to inactivate Salmonella in beef jerky. Decreasing meat pH significantly (p<0.05) increased the efficacy of PS and T to reduce the levels of Salmonella spp. Beef jerky processed at 82°C, pH5.5, with 0.25% PS to a final aw of 0.7 resulted in a maximum Salmonella logarithmic reduction of 5.0log10CFU/g and an inactivation rate of 1.3log10(CFU/g)/h. The predictive model developed can be used to effectively design drying processes for beef jerky under low humidity conditions and thereby, ensuring an adequate degree of protection against risks associated with Salmonella spp.

  1. Effect of pH, water activity and gel micro-structure, including oxygen profiles and rheological characterization, on the growth kinetics of Salmonella Typhimurium.

    PubMed

    Theys, T E; Geeraerd, A H; Verhulst, A; Poot, K; Van Bree, I; Devlieghere, F; Moldenaers, P; Wilson, D; Brocklehurst, T; Van Impe, J F

    2008-11-30

    In this study, the growth of Salmonella Typhimurium in Tryptic Soy Broth was examined at different pH (4.50-5.50), water activity a(w) (0.970-0.992) and gelatin concentration (0%, 1% and 5% ) at 20 degrees C. Experiments in TSB with 0% gelatin were carried out in shaken erlenmeyers, in the weak 1% gelatin media in petri plates and in the firm 5% gelatin media in gel cassettes. A quantification of gel strength was performed by rheological measurements and the influence of oxygen supply on the growth of S. Typhimurium was investigated. pH, as well as a(w) as well as gelatin concentration had an influence on the growth rate. Both in broth and in gelatinized media, lowering pH or water activity caused a decrease of growth rate. In media with 1% gelatin a reduction of growth rate and maximal cell density was observed compared to broth at all conditions. However, the effects of decreasing pH and a(w) were less pronounced. A further increase in gelatin concentration to 5% gelatin caused a small or no additional drop of growth rate. The final oxygen concentration dropped from 5.5 ppm in stirred broth to anoxic values in petri plates, also when 0% and 5% gelatin media were tested in this recipient. Probably, not stirring the medium, which leads to anoxic conditions, has a more pronounced effect on the growth rate of S. Typhimurium then medium solidness. Finally, growth data were fitted with the primary model of Baranyi and Roberts [Baranyi, J. and Roberts, T. A., 1994. A dynamic approach to predicting bacterial growth in food. International Journal of Food Microbiology 23, 277-294]. An additional factor was introduced into the secondary model of Ross et al. [Ross, T. and Ratkowsky, D. A. and Mellefont, L. A. and McMeekin, T. A., 2003. Modelling the effects of temperature, water activity, pH and lactic acid concentration on the growth rate of Escherichia coli. International Journal of Food Microbiology 82, 33-43.] to incorporate the effect of gelatin concentration, next to

  2. Automatic Weather Station (AWS) Lidar

    NASA Technical Reports Server (NTRS)

    Rall, Jonathan A.R.; Abshire, James B.; Spinhirne, James D.; Smith, David E. (Technical Monitor)

    2000-01-01

    An autonomous, low-power atmospheric lidar instrument is being developed at NASA Goddard Space Flight Center. This compact, portable lidar will operate continuously in a temperature controlled enclosure, charge its own batteries through a combination of a small rugged wind generator and solar panels, and transmit its data from remote locations to ground stations via satellite. A network of these instruments will be established by co-locating them at remote Automatic Weather Station (AWS) sites in Antarctica under the auspices of the National Science Foundation (NSF). The NSF Office of Polar Programs provides support to place the weather stations in remote areas of Antarctica in support of meteorological research and operations. The AWS meteorological data will directly benefit the analysis of the lidar data while a network of ground based atmospheric lidar will provide knowledge regarding the temporal evolution and spatial extent of Type la polar stratospheric clouds (PSC). These clouds play a crucial role in the annual austral springtime destruction of stratospheric ozone over Antarctica, i.e. the ozone hole. In addition, the lidar will monitor and record the general atmospheric conditions (transmission and backscatter) of the overlying atmosphere which will benefit the Geoscience Laser Altimeter System (GLAS). Prototype lidar instruments have been deployed to the Amundsen-Scott South Pole Station (1995-96, 2000) and to an Automated Geophysical Observatory site (AGO 1) in January 1999. We report on data acquired with these instruments, instrument performance, and anticipated performance of the AWS Lidar.

  3. Combined effects of weak acid preservatives, pH and water activity on growth of Eurotium species on a sponge cake.

    PubMed

    Guynot, M E; Ramos, A J; Sala, D; Sanchis, V; Marín, S

    2002-06-05

    The combined effects of weak acid preservatives (sorbate, benzoate and propionate), pH (6.0, 7.5) and water activity (a(w)) levels (0.80, 0.85, 0.90) on growth of four Eurotium species isolated from bakery products on a sponge cake analogue were studied. Even though it is universally known that these preservatives are much more effective at lower pH values, we chose a 6-7.5 level to correlate with the pH of the Spanish cake product studied. In general, 0.3% doses of all three preservatives were effective only when they were applied at pH 6.0 and at 0.80-0.85 a(w). Potassium sorbate was clearly the most effective in inhibiting growth of all isolates. Under the conditions tested, application of all three preservatives added at 0.03% acted as growth promoter of all isolates rather than having a preservative effect.

  4. Washing of the AW-101 entrained solids

    SciTech Connect

    GJ Lumetta

    2000-03-31

    BNFL Inc. (BNFL) is under contract with the US Department of Energy, River Protection Project (DOE-RPP) to design, construct, and operate facilities for treating wastes stored in the single-shell and double-shell tanks at the Hanford Site, Richland, Washington. The DOE-BNFL RPP contract identifies two feeds to the waste treatment plant: (1) primarily liquid low-activity waste (LAW) consisting of less than 2 wt% entrained solids and (2) high-level waste (HLW) consisting of 10 to 200 g/L solids slurry. This report describes the results of a test conducted by Battelle to assess the effects of inhibited water washing on the composition of the entrained solids in the diluted AW-101 low-activity waste (LAW) sample. The objective of this work was to gather data on the solubility of the AW-101 entrained solids in 0.01 M NaOH, so that BNFL can evaluate whether these solids require caustic leaching. The work was conducted according to test plan BNFL-TP-29953-9, Rev. 0, LAW Entrained Solids Water Wash and Caustic Leach Testing. The test went according to plan, with no deviations from the test plan. Based on the results of the 0.01 M NaOH washing, a decision was made by BNFL to not proceed with the caustic leaching test. The composition of the washed solids was such that caustic leaching would not result in significant reduction in the immobilized HLW volume.

  5. Improved granular activated carbon for the stabilization of wastewater PH

    SciTech Connect

    Farmer, R.W.; Dussert, B.W.; Kovacic, S.L.

    1996-12-31

    Laboratory studies have identified the cause of the pH rise, which occurs during water treatment with activated carbon, as an interaction between the naturally occurring anions and protons in the water and the carbon surface. The interaction can be described as an ion exchange type of phenomenon, in which the carbon surface sorbs the anions and corresponding hydronium ions from the water. These studies have shown that the anion sorption and resulting pH increase is independent of the raw material used for the activated carbon production, e.g. bituminous or subbituminous coal, peat, wood or coconut. Also, the pH excursions occur with virgin, reactivated, and acid washed granular carbons. Current pH control technologies focus on adjustment of the wastewater pH prior to discharge or recycle of the initial effluent water until the pH increase abates. However, improved water pH control options have been realized by altering the carbon surface through controlled oxidation rather than the water chemistry or extended preprocessing at the treatment site.

  6. Validated empirical models describing the combined effect of water activity and pH on the heat resistance of spores of a psychrotolerant Bacillus cereus strain in broth and béchamel sauce.

    PubMed

    Samapundo, S; Heyndrickx, M; Xhaferi, R; Devlieghere, F

    2011-10-01

    The major objective of this study was to evaluate and model the combined effect of the water activity (a(w)) and pH of the heating menstrum on the heat resistance of spores of a psychrotolerant Bacillus cereus strain isolated from béchamel sauce. Two models, a quadratic polynomial equation and a reparameterized function, were assessed for their ability to describe the combined influence of a(w) and pH on the D(85°C)-values of the B. cereus isolate in tryptone soy broth. The performance of the models was validated by challenging the models with data independently collected in broth and béchamel sauce. Both models were found to adequately describe the validation data obtained in broth. However, it was determined that in béchamel sauce the predictions of the polynomial function not only showed bias (bias factor = 1.156) but were also fail-dangerous, as they deviated from the validation data by 17.2%. The reparameterized function was determined to be a good predictor of the D(85°C)-values in béchamel sauce as it showed no bias (bias factor = 1.033) and its predictions differed by only 7.9% from the validation data. The reparameterized function can be used to provide estimates of the minimum processing conditions required to achieve desired levels of spore inactivation within the a(w) and pH ranges studied and to determine the potential changes in heat resistance of B. cereus spores when a(w) and pH are changed, for example, during product reformulation. As validation of heat resistance models is rarely performed, let alone in actual food products, the models evaluated and validated in this study (in particular the reparameterized function) are of immediate relevance to the food industry.

  7. Modulation of autophagic activity by extracellular pH.

    PubMed

    Xu, Teng; Su, Hang; Ganapathy, Suthakar; Yuan, Zhi-Min

    2011-11-01

    Reprogramming energy metabolism from oxidative phosphorylation to aerobic glycolysis, a common feature of human cancer, is associated with a relative acidic tumor microenvironment which can sometimes be further accentuated by hypoxia operating within most solid tumors. We found that alteration of extracellular pH induces marked and rapid changes of autophagic activity. Interestingly, acidic and basic conditions induced completely opposite effect on autophagy, with its activity suppressed at lower pH whereas stimulated at higher pH. Gene knockdown experiments indicated that pH induced-autophagy requires Beclin 1, Vps34 and Atg5, key components of the autophagy pathway. Of note, an acidic condition not only inhibits the basal but also blocks the starvation-induced autophagy activity. Significantly, examination of different areas of tumor mass revealed a lower autophagic activity within the inner region than the outer region. These findings have important implications on the connections between autophagy and cancer as well as a wide range of other physiological and pathological processes.

  8. Surface Treatment of Sonotrode Material Aw 7075

    NASA Astrophysics Data System (ADS)

    Emmer, Štefan; Baksa, Peter; Kováčik, Jaroslav

    2014-12-01

    Electro spark deposited TiB2 functional layer and composite functional layers Ni-TiB2 and INC713LC-TiB2 on the aluminium alloy substrate AW 7075 were evaluated with respect to achieved microhardness, roughness, and microstructure. The thicknesses of the layers were observed in the range of 30 to 50 microns. Hardness of the layers (HV) is several times higher than the hardness of the aluminium alloy substrate AW 7075.

  9. AIRS Observations of DomeC in Antarctica and Comparison with Automated Weather Stations (AWS)

    NASA Technical Reports Server (NTRS)

    Aumann, Hartmut H.; Gregorich, Dave; Broberg, Steve

    2006-01-01

    We compare the surface temperatures at Dome Concordia (DomeC) deduced from AIRS data and two Automatic Weather Stations at Concordia Station: AWS8989 , which has been in operation since December 1996, and AWS.it, for which data are available between January and November 2005. The AWS8989 readings are on average 3 K warmer than the AWS.it readings, with a warmer bias in the Antarctic summer than in the winter season. Although AIRS measures the skin brightness temperature, while the AWS reports the temperature of the air at 3 meter above the surface, the AIRS measurements agree well with the AWS.it readings for all data and separately for the summer and winter seasons, if data taken in the presence of strong surface inversions are filtered out. This can be done by deducing the vertical temperature gradient above the surface directly from the AIRS temperature sounding channels or indirectly by noting that extreme vertical gradients near the surface are unlikely if the wind speed is more than a few meters per second. Since the AIRS measurements are very well calibrated, the agreement with AWS.it is very encouraging. The warmer readings of AWS8989 are likely due to thermal contamination of the AWS8989 site by the increasing activity at Concordia Station. Data from an AWS.it quality station could be used for the evaluation of radiometric accuracy and stability of polar orbiting sounders at low temperatures. Unfortunately, data from AWS.it was available only for a limited time. The thermal contamination of the AWS8989 data makes long-term trends deduced from AWS8989 and possibly results about the rapid Antarctic warming deduced from other research stations on Antarctica suspect. AIRS is the first hyperspectral infrared sounder designed in support of weather forecasting and climate research. It was launched in May 2002 on the EOS Aqua spacecraft into a 704 km altitude polar sun-synchronous orbit. The lifetime of AIRS, estimated before launch to be at least 5 years is

  10. Influence of Water Activity on the Growth of Clostridium perfringens

    PubMed Central

    Strong, Dorothy H.; Foster, Edith F.; Duncan, Charles L.

    1970-01-01

    Each of four strains of Clostridium perfringens was grown in modified fluid thioglycolate medium which was adjusted to yield selected water activity (aw) levels. The adjustments to secure the desired aw levels were made with NaCl, KCl, or glucose. At each aw level, further modification was effected to produce four pH values. Cultures were incubated at either 37 or 46 C. The solute used to achieve the reduced aw levels appeared to have a definite effect on the magnitude of growth achieved, the rate of growth, and the limiting aw at which growth would occur. Use of glucose as the controlling solute permitted growth at the lowest aw level tested, 0.960, and yielded the greatest magnitude of growth as measured by turbidity values, at all of the aw levels investigated. Cultures grown in the medium with added KCl generally demonstrated the longest lag times and the least amount of growth. Regardless of specific solute used, as the aw level was lowered and the pH value decreased within each aw level, the rate and amount of growth were lessened. It appeared, however, that low pH values had less effect on inhibiting growth at low aw levels than at higher aw levels. Those cultures incubated at 46 C generally exhibited shorter lag periods than those at 37 C, although the maximal growth attained was somewhat less than that achieved at 37 C. The response to all of the investigated conditions was similar for each of the four strains tested. PMID:4318452

  11. AWS breaks new ground with soldering specification.

    SciTech Connect

    Vianco, Paul Thomas

    2008-08-01

    Joining technologies continue to advance with new materials, process innovations, and inspection techniques. An increasing number of high-valued, high-reliability applications -- from boilers and ship hulls to rocket motors and medical devices -- have required the development of industry standards and specifications in order to ensure that the best design and manufacturing practices are being used to produce safe, durable products and assemblies. Standards writing has always had an important role at the American Welding Society (AWS). The AWS standards and specifications cover such topics as filler materials, joining processes, inspection techniques, and qualification methods that are used in welding and brazing technologies. These AWS standards and specifications, all of which are approved by the American National Standards Institute (ANSI), have also provided the basis for many similar documents used in Europe and in Pacific Rim countries.

  12. Introduction to the Abrupt Wing Stall (AWS) Program

    NASA Technical Reports Server (NTRS)

    Hall, Robert M.; Woodson, Shawn H.

    2003-01-01

    The Abrupt Wing Stall (AWS) Program has addressed the problem of uncommanded, transonic lateral motions, such as wing drop, with experimental, computational, and simulation tools. Background to the establishment of the AWS program is given as well as program objectives. In order to understand the fundamental flow mechanisms that caused the undesirable motions for a pre-production version of the F/A-18E, steady and unsteady flow field details were gathered from dedicated transonic wind-tunnel testing and computational studies. The AWS program has also adapted a free-to-roll (FTR) wind-tunnel testing technique traditionally used for low-speed studies of lateral dynamic stability to the transonic flow regime. This FTR capability was demonstrated first in a proof-of -concept study and then applied to an assessment of four different aircraft configurations. Figures of merit for static testing and for FTR testing have been evaluated for two configurations that demonstrated wing drop susceptibility during full-scale flight conditions (the pre-production F/A-18E and the AV-8B at the extremes of its flight envelope) and two configurations that do not exhibit wing drop (the F/A-18C and the F-16C). Design insights have been obtained from aerodynamic computational studies of the four aircraft configurations and from computations quantifying the impact of the various geometric wing differences between the F/A-18C and the F/A-18E wings. Finally, the AWS program provides guidance for assessing, in the simulator, the impact of experimentally determined lateral activity on flight characteristics before going to flight.

  13. Introduction to the Abrupt Wing Stall (AWS) Program

    NASA Technical Reports Server (NTRS)

    Hall, Robert M.; Woodson, Shawn H.

    2003-01-01

    The Abrupt Wing Stall (AWS) Program has addressed the problem of uncommanded, transonic lateral motions, such as wing drop, with experimental, computational, and simulation tools. Background to the establishment of the AWS program is given as well as program objectives. In order to understand the fundamental flow mechanisms that caused the undesirable motions for a pre-production version of the F/A-18E, steady and unsteady flow field details were gathered from dedicated transonic wind-tunnel testing and computational studies. The AWS program has also adapted a free-to- roll (FTR) wind-tunnel testing technique traditionally used for low-speed studies of lateral dynamic stability to the transonic flow regime. This FTR capability was demonstrated first in a proof-of-concept study and then applied to an assessment of four different aircraft configurations. Figures of merit for static testing and for FTR testing have been evaluated for two configurations that demonstrated wing drop susceptibility during full-scale flight conditions (the pre-production F/A-l8E and the AV-8B at the extremes of its flight envelope) and two configurations that do not exhibit wing drop (the F/A-l8C and the F-l6C). Design insights have been obtained from aerodynamic computational studies of the four aircraft configurations and from computations quantifying the impact of the various geometric wing differences between the F/A-18C and the F/A-l8E wings. Finally, the AWS program provides guidance for assessing, in the simulator, the impact of experimentally determined lateral activity on flight characteristics before going to flight.

  14. Mark Twain and "The Awful German Language."

    ERIC Educational Resources Information Center

    Hedderich, Norbert

    2003-01-01

    Analyzes Mark Twain's 1869 essay "The Awful German Language" in terms of Twain's comments on morphological, syntactical, lexical, and phonological features of German. The topic is presented in the context of Twain's German language learning experience. Relevance of the article for German language instruction today is also described.…

  15. 75 FR 63088 - RIN 1018-AW77

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-14

    ... 1018-AW77 Subsistence Management Regulations for Public Lands in Alaska, Subpart B; Special Actions... of Subsistence Management, 1011 East Tudor Road, MS 121, Anchorage, AK 99503, or on the Office of Subsistence Management Web site ( http://alaska.fws.gov/asm/index.cfml ). FOR FURTHER INFORMATION...

  16. A polysaccharide isolated from Agaricus blazei Murill (ABP-AW1) as a potential Th1 immunity-stimulating adjuvant.

    PubMed

    Cui, Liran; Sun, Yongxu; Xu, Hao; Xu, Huiyu; Cong, Huan; Liu, Jicheng

    2013-10-01

    In the present study, a low molecular weight polysaccharide, ABP-AW1, isolated from Agaricus blazei Murill was assessed for its potential adjuvant activity. ABP-AW1 is considered to create a 'depot' of antigen at a subcutaneous injection site. ICR mice were immunized with 100 μg ovalbumin (OVA) alone or with 100 μg OVA formulated in 0.9% saline containing 200 μg aluminum (alum) or ABP-AW1 (50, 100 and 200 μg) on days 1 and 15. Two weeks after the secondary immunization, splenocyte proliferation, the expression of surface markers, cytokine production and the OVA-specific antibody levels in the serum were determined. The OVA/ABP-AW1 vaccine, in comparison with OVA alone, markedly increased the proliferation of splenic lymphocytes and elicited greater antigen-specific CD4(+) T cell activation, as determined by splenic CD4(+)CD69(+) T cells and Th1 cytokine interferon (IFN)-γ release. The combination of ABP-AW1 and OVA also enhanced IgG2b antibody responses to OVA. In conclusion, these data indicated that ABP-AW1 significantly enhanced the humoral and cellular immune responses against OVA in the mice, suggesting that ABP-AW1 stimulated Th1-type immunity. We suggest that ABP-AW1 may serve as a new adjuvant.

  17. [Phosphatase activity in Amoeba proteus at low pH].

    PubMed

    Sopina, V A

    2009-01-01

    In free-living Amoeba proteus (strain B), three forms of tartrate-sensitive phosphatase were revealed using PAGE of the supernatant of ameba homogenates obtained with 1% Triton X-100 or distilled water and subsequent staining of gels with 2-naphthyl phosphate as substrate (pH 4.0). The form with the highest mobility in the ameba supernatant was sensitive to all tested phosphatase activity modulators. Two other forms with the lower mobilities were completely or significantly inactivated not only by sodium L-(+)-tartrate, but also by L-(+)-tartaric acid, sodium orthovanadate, ammonium molybdate, EDTA, EGTA, o-phospho-L-tyrosine, DL-dithiotreitol, H2O2, 2-mercaptoethanol, and ions of heavy metals - Fe2+, Fe3+, and Cu2+. Based on results of inhibitory analysis, lysosome location in the ameba cell, and wide substrate specificity of these two forms, it has been concluded that they belong to nonspecific acid phosphomonoesterases (AcP, EC 3.1.3.2). This AcP is suggested to have both phosphomonoesterase and phosphotyrosyl-protein phosphatase activitis. Two ecto-phosphatases were revealed in the culture medium, in which amebas were cultivated. One of them was inhibited by the same reagents as the ameba tartrate-sensitive AcP and seems to be the AcP released into the culture medium in the process of exocytosis of the content of food vacuoles. In the culture medium, apart from this AcP, another phosphatase was revealed, which was not inhibited by any tested inhibitors of AcP and alkaline phosphatase. It cannot be ruled out that this phosphatase belong to the ecto-ATPases found in many protists; however, its ability to hydrolyze ATP has not yet been proven.

  18. Modelling the effect of temperature, pH, water activity, and organic acids on the germination time of Penicillium camemberti and Penicillium roqueforti conidia.

    PubMed

    Kalai, Safaa; Anzala, Lexane; Bensoussan, Maurice; Dantigny, Philippe

    2017-01-02

    In this study, the influence of environmental factors on the germination time of Penicillium camemberti and Penicillium roqueforti conidia was evaluated. To do so, the effects of i/temperature, pH, water activity, and ii/organic acids were determined using models based on i/cardinal values, and ii/minimum inhibitory concentration (MIC) respectively. Cardinal values for germination of conidia were not observed to be species dependent. Minimum temperatures were estimated to be below the freezing point, with an optimum of 26.9°C, and a maximum of 33.5°C. For both species, minimal and optimal aw values were found to be 0.83 and 0.99, respectively, while for pH these values corresponded to 2.9, and 5.6. MIC values could not be determined for lactic acid because conidia of both species germinated in up to 1M concentrations, the highest concentration tested. At pH5.6, P. camemberti (MIC=0.197M) was more sensitive to propionic acid than P. roqueforti (MIC=0.796M).

  19. Low pH effects on swimming activity of Ambystoma salamander larvae

    SciTech Connect

    Kutka, F.J. . Dept. of Animal Ecology)

    1994-11-01

    Swimming activity of larval Ambystoma laterale increased linearly with pH between pH 4.0 and 6.5; near inactivity occurred at pH 4.0. Ambystoma maculatum exhibited a weaker relationship between activity and pH, and overall was less active than A. laterale. However, survival of larval A. maculatum declined linearly with pH in the presence of adult diving beetles (Dytiscus verticalis), with significantly lower survival below pH 4.8. These results suggest that, as with embryos, salamander larvae may suffer from pH levels below 5.0, even though these levels are not directly lethal. Because of their sensitivity, ease, and short duration, activity tests are recommended with amphibian larvae for use in other risk assessments.

  20. Effect of environmental pH on enzyme activity and growth of Bacteroides gingivalis W50.

    PubMed Central

    McDermid, A S; McKee, A S; Marsh, P D

    1988-01-01

    Since the pH of the gingival crevice increases from below neutrality in health to above pH 8 in disease, we decided to investigate the effect of environmental pH on the growth and enzyme activity of Bacteroides gingivalis W50. Cells were grown in a chemostat under hemin-excess conditions over a range of pH values; stable growth was observed only between pH 6.7 and 8.3, with the maximum yields obtained between pH 7.0 and 8.0. The enzyme profile of cells varied markedly with pH. Enzymes with a specificity for gingival connective tissue (collagenase, hyaluronidase) were produced optimally at or below neutral pH, whereas trypsinlike activity increased with the growth pH and was maximal at pH 8.0. Chymotrypsinlike activity was generally low, although its activity was highest at the extremes of growth pH, i.e., at pH 6.7 and 8.3. Inhibitor studies provided evidence that the breakdown of collagen involved the concerted action of both a collagenase and the trypsinlike enzyme. The ratio of trypsin to collagenolytic activity rose from 1:1 during growth at neutral pH and below to almost 7:1 during growth at pH 8.3. Thus B. gingivalis appears to be uniquely adapted as a periodontopathic organism in that under environmental conditions likely to prevail during the initial stages of pocket development it produces maximally those enzymes with a tissue-damaging potential. Then, as the pH of the pocket rises during the host inflammatory response, the activity of the trypsinlike enzyme increases markedly, which may enable cells to inactivate key components of the host defenses such as immunoglobulins and complement. PMID:3281900

  1. [Professor Wacław Kuśnierczyk (1908-1997)--Pro Memoria in the century of birthday].

    PubMed

    Brozek, Krzysztof; Kozakiewicz, Jacek; Kierzek, Andrzej

    2009-01-01

    Wacław Kuśnierczyk was born in 1908 in Sniatyń. He received the degree in medicine at Jan Kazimierz University in Lwów in 1932. He did his PhD degree under Professor Zaleski supervision in 1938 at Jan Kazimierz University. At that time he concentrated his scientific activity on research on tuberculosis. In 1953 he obtained the title of second degree specialist in ear, nose and throat diseases. He became a chief of Otolaryngology at Urban Hospital No 4 in Katowice in 1960. Since then this eminent physician was working on tumours located in upper respiratory tract and the possibility of its endoscopic diagnosis at Silesian Academy of Medicine in Katowice. As one of the first he pointed out the negative influence of smoking cigarettes on cancer of larynx. It was Wacław Kuśnierczyk who implemented new priorities for integrated programs in patient care, research, education and cancer prevention. He has published widely in peer reviewed journals and has edited or contributed to many books. He has given many major lectures and is the recipient of numerous prestigious awards for his scientific accomplishments. The achievement of Professor Kuśnierczyk were the valuable source of information for the physicians. In 1997, on the 31st of January he died in Katowice.

  2. Activity and stability trends of perovskite oxides for oxygen evolution catalysis at neutral pH.

    PubMed

    Han, Binghong; Risch, Marcel; Lee, Yueh-Lin; Ling, Chen; Jia, Hongfei; Shao-Horn, Yang

    2015-09-21

    Perovskite oxides (ABO3) have been studied extensively to promote the kinetics of the oxygen evolution reaction (OER) in alkaline electrolytes. However, developing highly active catalysts for OER at near-neutral pH is desirable for many photoelectrochemical/electrochemical devices. In this paper, we systematically studied the activity and stability of well-known perovskite oxides for OER at pH 7. Previous activity descriptors established for perovskite oxides at pH 13, such as having an eg occupancy close to unity or having an O p-band center close to Fermi level, were shown to scale with OER activity at pH 7. Stability was a greater challenge at pH 7 than at pH 13, where two different modes of instability were identified from combined transmission electron microscopy and density functional theory analyses. Perovskites with O p-band close to Fermi level showed leaching of A-site atoms and surface amorphization under all overpotentials examined at pH 7, while those with O p-band far from Fermi level were stable under low OER current/potential but became unstable at high current/potential accompanied by leaching of B-site atoms. Therefore, efforts are needed to enhance the activity and stability of perovskites against A-site or B-site loss if used at neutral pH.

  3. Phosphoglycolate phosphatase. Effect of cation and pH on activity.

    PubMed

    Christeller, J T; Tolbert, N E

    1978-03-25

    P-glycolate phosphatase requires divalent cations for activity. Activity-pH curves identified 2 active site residues with pK values at pH 5.7 and pH 9.1 in the presence of magnesium and at pH 5.7 and pH 7.5 in the presence of manganese or cobalt. Saturation velocity kinetics enabled the identification of two distinct divalent cation binding sites. The first, nonspecific site has a K0.5 of 2 to 7 x 10(-5) M, depending on the cation and the pH. The second site, which is specific for magnesium, binds this cation in a negatively cooperative fashion. The affinity at pH 8.1 varies approximately 100-fold from the first magnesium bound to the fourth. The negative cooperativity is greatest at high pH. Because the pH range of activity is very broad, both the phosphate monoanion and dianion of P-glycolate must be bound as the substrate. The concentration of these two species at the apparent Km is independent of magnesium concentration. The P-glycolate.magnesium complex is kinetically inactive.

  4. Influence of pH, water activity and acetic acid concentration on Listeria monocytogenes at 7 degrees C: data collection for the development of a growth/no growth model.

    PubMed

    Vermeulen, A; Gysemans, K P M; Bernaerts, K; Geeraerd, A H; Van Impe, J F; Debevere, J; Devlieghere, F

    2007-03-20

    Growth/no growth models can be used to determine the chance that microorganisms will grow in specific environmental conditions. As a consequence, these models are of interest in the assessment of the safety of foods which can be contaminated with food pathogens. In this paper, growth/no growth data for Listeria monocytogenes (in a monoculture and in a mixed strain culture) are presented. The data were gathered at 7 degrees C in Nutrient Broth with different combinations of environmental factors pH (5.0-6.0, six levels), water activity (0.960-0.990, six levels) and acetic acid concentration (0-0.8% (w/w), five levels). This combination of environmental factors for the development of a growth/no growth model was based on the characteristics of sauces and mayonnaise based salads. The strains used were chosen from screening experiments in which the pH, water activity and acetic acid resistance of 26 L. monocytogenes strains (LFMFP culture collection) was determined at 30 degrees C in Brain Heart Infusion broth. The screening showed that most L. monocytogenes strains were not able to grow at a(w)<0.930, pH<4.3 or a total acetic acid concentration >0.4% (w/w). Among these strains, the ones chosen were the most resistant to one of these factors in the hope that, if the resulting model predicted no growth at certain conditions for those more resistant strains, then these predictions would also be valid for the less resistant strains. A mixed strain culture was also examined to combine the strains that were most resistant to one of the factors. A full factorial design with the selected strains was tested. The experiments were performed in microtiter plates and the growth was followed by optical density measurements at 380 nm. The plates were inoculated with 6 log CFU/ml and twenty replicates were made for each treatment combination. These data were used (1) to determine the growth/no growth boundary and (2) to estimate the influence of the environmental conditions on the

  5. Comparison of AIRS and IASI Surface Observations of DomeC in Antarctica with Surface Temperatures Reported by AWS8989

    NASA Astrophysics Data System (ADS)

    Elliott, D. A.; Aumann, H. H.

    2008-12-01

    The decrease of the ice in the Antarctic indicates that the land and the ocean along the coastline are warming up. Representative numbers for warming at the surface further inland are much more complicated due to the vast size of the continent. The Automated Weather Station AWS8989 has been reporting temperatures from Concordia Station on DomeC in Antarctica every 10 minutes since 1996. AWS8989 is located about 1 mile from the power plant at Concordia Station. We compare the surface temperatures at DomeC deduced from Atmospheric Infrared Sounder (AIRS) and Infrared Atmospheric Sounding Interferometer (IASI) data to the surface temperature reported by Automated Weather Station AWS8989 for the year between May 1, 2007 and April 30, 2008. AIRS and IASI measure the mean skin brightness temperature in a 50-km-radius circle from DomeC, while the AWS reports the temperature of the air at 3 meters above the surface. The AIRS and IASI measurements agree within 50 mK over the entire temperature range from 190 K in the winter to 245 K in the summer, but consistently report a colder temperature than the AWS8989. The warm bias of AWS8989 is season dependent, changing from 1.5 K warm in the winter to 5.5 K warm in the summer. Comparison of AIRS data in 2005 with a temporary Italian AWS (Aumann et al. 2006) and located several miles upwind from the power station, showed no significant temperature bias throughout the year 2005. The warm readings of AWS8989 are likely due the combination of a season-independent 1.5 K warm calibration bias in the AWS8989 sensor plus thermal contamination of the AWS8989 site. This heat island effect ranges from near zero during the low-activity winter months to about 4 K during the summer months with the highest activity at Concordia Station. The fact that activities at DomeC are increasing makes surface temperature trends from AWS8989 suspect. AIRS and IASI are hyperspectral infrared sounders designed in support of weather forecasting and climate

  6. [Ludwik Hirszfeld (1884-1954) the Wrocław Years, 1945-1954].

    PubMed

    Heimrath, T

    2000-01-01

    Beginning with an initial overview of his earlier activities as a scholar, this article is a study of the years which Ludwik Hirszfeld spent in Wrocław and is based on published sources as well as the reminiscences of both his colleagues and of this author. Hirszfeld himself, in his described his life autobiography "History of One Life" until just after the Second World War. The years he spent in Wrocław organizing the Medical Department, of which he was the first director from 1945-1946 were unusually active ones. Hirszfeld was also at that time founder of the Department of Medical Microbiology and later of the Institute for Immunology and Experimental Therapy of the Polish Academy of Sciences in Wrocław. He was simultaneously involved in highly successful scientific research particularly in the area of immunopathology during pregnancy. He was a popular lecturer on this subject with a special gift for reaching his audience as well as an active participant in scientific discussions in a variety of foral forums. He had a great personal influence on the first generation of staff and students of the Medical Department in Wrocław in the years immediately following the end to the Second World War.

  7. Improved granular activated carbon for the stabilization of wastewater pH

    SciTech Connect

    1996-10-01

    Many times the start up of granular activated carbon adsorption systems for the control of organic contaminants in wastewater cm exhibit unacceptable increases in the adscurber effluent pH. Experience shows that the duration of the pH increase ranges from several hours to several days, during which time several hundred bed volumes of water can be discharged with a pH in excess of 9. Laboratory studies have identified the cause of the pH rise as an interaction between the naturally occurring anions and protons ar the water and the carbon surface. The interaction can be described as an ion exchange type of phenomenon, in which the carbon surface sorbs the anions and corresponding hydronium ions from the water. Capacities of the carbon for the anions range from 2 to 9 mg/g GAC, depending upon the water characteristics, the carbon type, the nature of the anion and its influent concentration. These studies have shown de the anion sorption and resulting pH increase is independent of the raw material used for die activated carbon production, e.g. bituminous or sub-bituminous coal, peat, wood or coconut. Also, the pH excursions occur with virgin, reactivated, and acid washed granular carbons. Current pH control technologies focus on adjustment of wastewater pH prior to discharge or recycle of the initial effluent water until the pH increase abates. However, improved water pH control options have been realized by altering the carbon surface rather than the water chemistry. The change to the carbon surface is accomplished through a controlled oxidation process. This process provides a more acidic carbon surface with a reduced affinity for the anions in the waste water. As a result, the pH excursions above 9 are eliminated and the initial effluent from the adsorption system can be discharged without further treatment.

  8. pH at the Air-Ice Interface: Connections to Halogen Activation Chemistry

    NASA Astrophysics Data System (ADS)

    Wren, S. N.; Abbatt, J.; Donaldson, D. J.

    2014-12-01

    Halogen activation - the conversion of relatively inert sea-salt derived halides, into reactive halogen species - has significant implications for perturbing the oxidative capacity of the boundary layer, as well as for mercury oxidation. Developing a good, mechanistic understanding of halogen activation chemistry - and particularly its pH dependence - requires knowledge of surface pH. Here we described the development and use of a surface-sensitive spectroscopic approach to investigate pH at frozen surfaces. The approach, which involves glancing-angle laser-induced fluorescence in conjunction with pH-sensitive fluorescent dyes (acridine and harmine), was used to study pH at frozen surfaces following the deposition of gas phase acids or bases. We demonstrate that frozen 'pure water' and frozen 'salt water' surfaces exhibit different responses. The responses are consistent with the presence of a chemically unique, disordered region at the frozen 'pure water' surface and the presence of liquid brine at the frozen 'salt water' surface. Significantly, we show that a frozen sea water surface is resistant to changes in pH, suggesting that some buffering capacity is maintained upon freezing. Finally the pH dependence of bromine and chlorine activation from artificial saline snow is presented. Bromine release is found to depend less strongly on pre-freezing snow pH, but the release of both species is favoured for low pre-freezing pH values. In the context of these pH and halogen activation studies, the potential importance of various frozen sea ice substrates for promoting halogen activation chemistry will be discussed.

  9. Thiourea compound AW00178 sensitizes human H1299 lung carcinoma cells to TRAIL-mediated apoptosis.

    PubMed

    Ryu, Byung Jun; Hwang, Mi-Kyung; Park, Mikyung; Lee, Kyunghee; Kim, Seong Hwan

    2012-06-15

    The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) triggers apoptosis in a wide variety of cancer cells. Recently, cancer cell resistance to TRAIL-mediated apoptosis has become a challenging issue in the development of TRAIL-based anti-cancer therapies. In this study, we found that 1-(5-chloro-2-methyl-phenyl)-3-[4-(5-trifluoromethyl-pyrazol-1-yl)-phenyl]-thiourea (AW00178) was able to sensitize TRAIL-resistant human lung cancer H1299 cells to TRAIL-mediated apoptosis. Treatment with AW00178, either alone or in combination with TRAIL, induced the expression of CHOP, a protein related to TRAIL sensitivity, and reduced the expression of survivin, an anti-apoptotic protein involved in TRAIL resistance. Additionally, AW00178, alone or in combination with TRAIL, induced the activation of c-Jun and inactivation of Akt. A pharmacologic inhibition study revealed that c-Jun activation and Akt inactivation were strongly related to CHOP induction and survivin down-regulation, respectively. In summary, these results suggested that AW00178 mediated sensitization to TRAIL-mediated apoptosis in H1299 cells by increasing sensitivity and decreasing resistance to TRAIL via the induction of c-Jun-dependent CHOP expression and the reduction of Akt-dependent survivin expression, respectively.

  10. Antitumour, antimicrobial and catalytic activity of gold nanoparticles synthesized by different pH propolis extracts

    NASA Astrophysics Data System (ADS)

    Gatea, Florentina; Teodor, Eugenia Dumitra; Seciu, Ana-Maria; Covaci, Ovidiu Ilie; Mănoiu, Sorin; Lazăr, Veronica; Radu, Gabriel Lucian

    2015-07-01

    The Romanian propolis was extracted in five different media, respectively, in water (pH 6.8), glycine buffer (pH 2.5), acetate buffer (pH 5), phosphate buffer (pH 7.4) and carbonate buffer (pH 9.2). The extracts presented different amounts of flavonoids and phenolic acids, increasing pH leading to higher concentrations of active compounds. Five variants of gold nanoparticles suspensions based on different pH Romanian propolis aqueous extracts were successfully synthesized. The obtained nanoparticles presented dimensions between 20 and 60 nm in dispersion form and around 18 nm in dried form, and different morphologies (spherical, hexagonal, triangular). Fourier transform infrared spectroscopy proved the attachment of organic compounds from propolis extracts to the colloidal gold suspensions and X-ray diffraction certified that the suspensions contain metallic gold. The obtained propolis gold nanoparticles do not exhibit any antibacterial or antifungal activity, but presented different catalytic activities and toxicity on tumour cells.

  11. pH and Antimicrobial Activity of Portland Cement Associated with Different Radiopacifying Agents.

    PubMed

    Guerreiro-Tanomaru, Juliane Maria; Cornélio, Ana Lívia G; Andolfatto, Carolina; Salles, Loise P; Tanomaru-Filho, Mário

    2012-01-01

    Objective. The aim of this study was to evaluate the antimicrobial activity and pH changes induced by Portland cement (PC) alone and in association with radiopacifiers. Methods. The materials tested were pure PC, PC + bismuth oxide, PC + zirconium oxide, PC + calcium tungstate, and zinc oxide and eugenol cement (ZOE). Antimicrobial activity was evaluated by agar diffusion test using the following strains: Micrococcus luteus, Streptococcus mutans, Enterococcus faecalis, Pseudomonas aeruginosa, and Candida albicans. After 24 hours of incubation at 37°C, inhibition of bacterial growth was observed and measured. For pH analysis, material samples (n = 10) were placed in polyethylene tubes and immersed in 10 mL of distilled water. After 12, 24, 48, and 72 hours, the pH of the solutions was determined using a pH meter. Results. All microbial species were inhibited by the cements evaluated. All materials composed of PC with radiopacifying agents promoted pH increase similar to pure Portland cement. ZOE had the lowest pH values throughout all experimental periods. Conclusions. All Portland cement-based materials with the addition of different radiopacifiers (bismuth oxide, calcium tungstate, and zirconium oxide) presented antimicrobial activity and pH similar to pure Portland cement.

  12. Applied PhD Research in a Work-Based Environment: An Activity Theory-Based Analysis

    ERIC Educational Resources Information Center

    Granata, S. N.; Dochy, F.

    2016-01-01

    Activity theory is used to compare PhD undertaken at university, that is, academic PhD, with PhD performed in collaboration with industry, that is, semi-industrial PhD. The research is divided into a literature review and a case study. Semi-industrial and academic PhD are modelled as activity systems, and differences are highlighted in terms of…

  13. Global lightning activity and climate change. Ph.D. Thesis

    SciTech Connect

    Price, C.G.

    1993-01-01

    The relationship between global lightning frequencies and global climate change is examined in this thesis. In order to study global impacts of climate change, global climate models or General Circulations Models (GCM`s) need to be utilized. Since these models have coarse resolutions many atmospheric phenomena that occur at subgrid scales, such as lightning, need to be parameterized whenever possible. The first chapter introduces a simple parameterization used to simulate total (intracloud and cloud-to-ground) lightning frequencies. The parameterization uses convective cloud top height to approximate lightning frequencies. The second chapter deals with a parameterization for simulating cloud-to-ground (CG) lightning around the globe. This parameterization uses the thickness of the cold cloud sector in thunderstorms (0 C to cloud top) to calculate the proportion of CG flashes in a particular thunderstorm. The third chapter deals with the modelling of lightning in the Goddard Institute for Space Studies (GISS) GCM. This chapter presents results from the model`s control run. The fourth chapter presents two climate change scenarios. One for a climate where the solar constant is reduced by 2% (5.9 C global cooling), and one for a climate with twice the present concentration of CO2 in the atmosphere (4.2 C global warming). The results imply a 24% / 30% decrease/increase in global lightning frequencies for the cooler/warmer climate. The fifth chapter considers the possibility of using the above findings to monitor future global warming. The results show that the earth`s ionospheric potential, which is regulated by global thunderstorm activity, could possibly supply valuable information regarding global surface temperature fluctuations. The sixth and final chapter looks at the implications of changes in both lightning frequencies and the hydrological cycle, as a result of global warming, on natural forest fires.

  14. The orbit of the Cepheid AW Per

    NASA Technical Reports Server (NTRS)

    Evans, Nancy Remage; Welch, Douglas L.

    1988-01-01

    An orbit for the classical Cepheid AW Per was derived. Phase residuals from the light curve are consistent with the light-time effect from the orbit. The companion was studied using IUE spectra. The flux distribution from 1300 to 1700 A is unusual, probably an extreme PbSi star, comparable to a B7V or B8V star. The flux of the composite spectrum from 1200 A through V is well matched by F7Ib and B8V standard stars with Delta M(sub upsilon) = 3(m) multiplied by 1. The mass function from the orbit indicates that the mass of the Cepheid must be greater that 4.7 solar mass if it is the more massive component. A B7V to B8V companion is compatible with the 1 sigma lower limit (3.5 solar mass) from the mass function. This implies that the Cepheid has the same mass, but the large magnitude difference rules this out. It is likely that the companion is itself a binary.

  15. An alkaline active xylanase: insights into mechanisms of high pH catalytic adaptation.

    PubMed

    Mamo, Gashaw; Thunnissen, Marjolein; Hatti-Kaul, Rajni; Mattiasson, Bo

    2009-09-01

    The alkaliphilic bacterium, Bacillus halodurans S7, produces an alkaline active xylanase (EC 3.2.1.8), which differs from many other xylanases in being operationally stable under alkaline conditions as well as at elevated temperature. Compared to non-alkaline active xylanases, this enzyme has a high percent composition of acidic amino acids which results in high ratio of negatively to positively charged residues. A positive correlation was observed between the charge ratio and the pH optima of xylanases. The recombinant xylanase was crystallized using a hanging drop diffusion method. The crystals belong to the space group P2(1)2(1)2(1) and the structure was determined at a resolution of 2.1 A. The enzyme has the common eight-fold TIM-barrel structure of family 10 xylanases; however, unlike non-alkaline active xylanases, it has a highly negatively charged surface and a deeper active site cleft. Mutational analysis of non-conserved amino acids which are close to the acid/base residue has shown that Val169, Ile170 and Asp171 are important to hydrolyze xylan at high pH. Unlike the wild type xylanase which has optimum pH at 9-9.5, the triple mutant xylanase (V169A, I170F and D171N), which was constructed using sequence information of alkaline sensitive xylanses was optimally active around pH 7. Compared to non-alkaline active xylanases, the alkaline active xylanases have highly acidic surfaces and fewer solvent exposed alkali labile residues. Based on these results obtained from sequence, structural and mutational analysis, the possible mechanisms of high pH stability and catalysis are discussed. This will provide useful information to understand the mechanism of high pH adaptation and engineering of enzymes that can be operationally stable at high pH.

  16. Pyrazinamide Is Active against Mycobacterium tuberculosis Cultures at Neutral pH and Low Temperature.

    PubMed

    den Hertog, Alice L; Menting, Sandra; Pfeltz, Richard; Warns, Matthew; Siddiqi, Salman H; Anthony, Richard M

    2016-08-01

    For the past decades, an acidic pH has been used to render Mycobacterium tuberculosis susceptible to pyrazinamide for in vitro testing. Here, we show that at the standard breakpoint concentration and reduced culture temperatures, pyrazinamide (PZA) is active against tuberculosis (TB) at neutral pH. This finding should help unravel the mechanism of action of PZA and allow drug susceptibility testing (DST) methods to be optimized.

  17. Active transport, ion movements, and pH changes : I. The chemistry of pH changes.

    PubMed

    Good, N E

    1988-10-01

    The transport of substances across cell membranes may be the most fundamental activity of living things. When the substance transported is any ion there can be a change in the concentration of hydrogen ions on the two sides of the membrane. These hydrogen ion concentration changes are not caused by fluxes of hydrogen ions although fluxes of hydrogen ions may sometimes be involved. The reason for the apparent contradiction is quite simple. All aqueous systems are subject to two constraints: (1) to maintain the charge balance, the sum of the cationic charges must equal the sum of the anionic charges and (2) the product of the molar concentration of H(+) and the molar concentration of OH(-), established and maintained by the association and the dissociation of water, remains always at 10(-14). As a consequence the concentrations of H(+) and OH(-) are determined uniquely by differences between the concentrations of the other cations and anions, with [H(+)] and [OH(-)] being dependent variables. Hydrogen ions and hydroxyl ions can be produced or consumed in local reactions whereas any strong ions such as Cl(-), Mg(2+), or K(+) can be neither produced nor consumed in biological reactions. Further consequences of these truisms are outlined here in terms of the chemistry of the kinds of reactions which can lead to pH changes.

  18. Effect of pH on DNA alkylation by enzyme-activated mitomycin C and porfiromycin.

    PubMed

    Yu, F; Pan, S S

    1993-06-01

    DNA adduct formation by enzyme-activated antibiotics, mitomycin C (MMC) or porfiromycin (PFM), at pH 7.6 or pH 6.0 under anaerobic conditions was analyzed by a 32P-postlabeling method. Antibiotic activation by rat liver NADPH-cytochrome P-450 reductase (EC 1.6.2.4) and bovine milk xanthine oxidase (EC 1.2.3.2) produced similar results. Five 32P-labeled MMC adducts were separated by thin layer chromatography and high performance liquid chromatography from DNA alkylated at either pH. Four of the radioactive spots separated by thin layer chromatography were identified as two monofunctional monoadducts [1" alpha and 1" beta forms of N2-(2" beta,7"-diaminomitosen-1"-yl)-2'-deoxyguanylic acid], one bifunctional monoadduct [N2-(10"-decarbamoyl-2",7"-diaminomitosen-1" alpha-yl)-2'-deoxyguanylic acid], and one cross-linked adduct [N2-(2" beta,7"-diamino-10"-deoxyguanyl-N2-yl-mitosen- 1" alpha-yl)-2'-deoxyguanylic acid]. One minor radioactive spot was not identified. By comparing DNA alkylated at the two pH values, based on equal amounts of 32P radioactivity, similar amounts of cross-links were detected. However, the DNA showed different ratios of the alpha and beta isomers of the monofunctional monoadduct. Furthermore, the DNA alkylated at pH 6.0 showed more bifunctional monoadducts than did the DNA alkylated at pH 7.6. Analysis of alkylated DNA by enzyme-activated PFM showed a similar spectrum of DNA adduct formation. The effect of pH on the distribution of the five PFM-DNA adducts was similar to that observed for the five MMC-DNA adducts. The distribution of adducts in DNA alkylated at the same pH was similar irrespective of which enzyme activated MMC or PFM. The pH of the reaction during DNA and MMC interaction was the determining factor for the quantitative distribution of the adducts. This pH effect may be important for the cytotoxicity of MMC and PFM in tumor cells that have high levels of reductive enzymes with low optimal pH values.

  19. Cross bridge-dependent activation of contraction in cardiac myofibrils at low pH.

    PubMed

    Swartz, D R; Zhang, D; Yancey, K W

    1999-05-01

    Striated muscle contracts in the absence of calcium at low concentrations of MgATP ([MgATP]), and this has been termed rigor activation because rigor cross bridges attach and activate adjacent actin sites. This process is well characterized in skeletal muscle but not in cardiac muscle. Rigor cross bridges are also thought to increase calcium binding to troponin C and play a synergistic role in activation. We tested the hypothesis that cross bridge-dependent activation results in an increase in contractile activity at normal and low pH values. Myofibrillar ATPase activity was measured as a function of pCa and [MgATP] at pH 7.0, and the data showed that, at pCa values of >/=5.5, there was a biphasic relationship between activity and [MgATP]. Peak activity occurred at 10-50 microM MgATP, and [MgATP] for peak activity was lower with increased pCa. The ATPase activity of rat cardiac myofibrils as a function of [MgATP] at a pCa of 9.0 was measured at several pH levels (pH 5.4-7.0). The ATPase activity as a function of [MgATP] was biphasic with a maximum at 8-10 microM MgATP. Lower pH did not result in a substantial decrease in myofibrillar ATPase activity even at pH 5.4. The extent of shortening, as measured by Z-line spacing, was greatest at 8 microM MgATP and less at both lower and higher [MgATP], and this response was observed at all pH levels. These studies suggest that the peak ATPase activity associated with low [MgATP] was coupled to sarcomere shortening. These results support the hypothesis that cross bridge-dependent activation of contraction may be responsible for contracture in the ischemic heart.

  20. β-Galactoside-binding activity of human galectin-1 at basic pH

    NASA Astrophysics Data System (ADS)

    Hiramatsu, Hirotsugu; Takeuchi, Katsuyuki; Fukuda, Koki; Nishino, Tomohide

    2013-06-01

    β-Galactoside-binding activity of human galectin-1 (hGal-1) was evaluated at pH 7-9.5 by fluorescence spectroscopy from the fraction bound to lactose gel (Y) and the lactose binding constant (Kb). Y decreases at pH > 8.2 ± 0.1 in the absence of NaCl, while it is constant in the presence of 150 mM NaCl. On the other hand, Kb is independent of pH and the NaCl concentration at basic pH. Analysis of Raman spectrum has shown that the pKa of Cys residues of hGal-1 is 8.5 ± 0.1 on average, indicating that about 40% of the six Cys residues of hGal-1 would be deprotonated at pH 8.2. The pH dependence of Y is explained by an increase of Coulombic repulsion among negatively charged hGal-1 on the lactose gel surface. This result suggests that Y is not always a good indicator of the β-galactoside-binding activity of galectins, which contain many Cys residues.

  1. The entomopathogenic fungus Metarhizium anisopliae alters ambient pH, allowing extracellular protease production and activity.

    PubMed

    St Leger, R J; Nelson, J O; Screen, S E

    1999-10-01

    Ambient pH regulates the expression of virulence genes of Metarhizium anisopliae, but it was unknown if M. anisopliae can regulate ambient pH. Mutants of M. anisopliae altered in production of oxalic acid were evaluated for the interrelationship of ambient pH, buffering capacity added to media, growth, and generation of extracellular proteases and ammonia. Wild-type and acid-overproducing mutants [Acid(+)] grew almost as well at pH 8 as at pH 6, but acid-non-producing [Acid(-)] mutants showed limited growth at pH 8, indicating that acid production is linked to the ability to grow at higher pH. Production of ammonia by M. anisopliae was strongly stimulated by low levels of amino acids in the medium when cells were derepressed for nitrogen and carbon. Likewise, although Aspergillus fumigatus and Neurospora crassa produced some ammonia in minimal media, addition of low levels of amino acids enhanced production. Ammonia production by A. fumigatus, N. crassa and M. anisopliae increased the pH of the medium and allowed production of subtilisin proteases, whose activities are observed only at basic pH. In contrast, protease production by the Acid(+) mutants of M. anisopliae was greatly reduced because of the acidification of the medium. This suggests that alkalinization by ammonia production is adaptive by facilitating the utilization of proteinaceous nutrients. Collectively, the data imply that ammonia may have functions related to regulation of the microenvironment and that it represents a previously unconsidered virulence factor in diverse fungi with the potential to harm tissues and disturb the host's immune system.

  2. Expanding the active pH range of Escherichia coli glutamate decarboxylase by breaking the cooperativeness.

    PubMed

    Thu Ho, Ngoc Anh; Hou, Chen Yuan; Kim, Woo Hyun; Kang, Taek Jin

    2013-02-01

    Bacterial glutamate decarboxylase (GAD) transforms glutamate into γ-aminobutyric acid (GABA) with the consumption of a proton. The enzyme is active under acidic environments only and sharply loses its activity as pH approaches neutrality with concomitant structural deformation. In an attempt to understand better the role of this cooperative loss of activity upon pH shifts, we prepared and studied a series of GAD site-specific mutants. In this report, we show that the cooperativeness was kept intact by at least two residues, Glu89 and His465, of which Glu89 is newly identified to be involved in the cooperativity system of GAD. Double mutation on these residues not only broke the cooperativity in the activity change but also yielded a mutant GAD that retained the activity at neutral pH. The resulting mutant GAD that was active at neutral pH inhibited the cell growth in a glycerol medium by converting intracellular Glu into GABA in an uncontrolled manner, which explains in part why the cooperativeness of GAD has to be kept by several layers of safety keepers. This unexpected result might be utilized to convert a low-valued by-product of biodiesel production, glycerol, into value-added product, GABA.

  3. Membrane-Associated Transporter Protein (MATP) Regulates Melanosomal pH and Influences Tyrosinase Activity

    PubMed Central

    Bin, Bum-Ho; Bhin, Jinhyuk; Yang, Seung Ha; Shin, Misun; Nam, Yeon-Ju; Choi, Dong-Hwa; Shin, Dong Wook; Lee, Ai-Young; Hwang, Daehee; Cho, Eun-Gyung; Lee, Tae Ryong

    2015-01-01

    The SLC45A2 gene encodes a Membrane-Associated Transporter Protein (MATP). Mutations of this gene cause oculocutaneous albinism type 4 (OCA4). However, the molecular mechanism of its action in melanogenesis has not been elucidated. Here, we discuss the role of MATP in melanin production. The SLC45A2 gene is highly enriched in human melanocytes and melanoma cell lines, and its protein, MATP, is located in melanosomes. The knockdown of MATP using siRNAs reduced melanin content and tyrosinase activity without any morphological change in melanosomes or the expression of melanogenesis-related proteins. Interestingly, the knockdown of MATP significantly lowered the melanosomal pH, as verified through DAMP analysis, suggesting that MATP regulates melanosomal pH and therefore affects tyrosinase activity. Finally, we found that the reduction of tyrosinase activity associated with the knockdown of MATP was readily recovered by copper treatment in the in vitro L-DOPA oxidase activity assay of tyrosinase. Considering that copper is an important element for tyrosinase activity and that its binding to tyrosinase depends on melanosomal pH, MATP may play an important role in regulating tyrosinase activity via controlling melanosomal pH. PMID:26057890

  4. Sodium-dependent pH regulation in active sea urchin sperm.

    PubMed

    Bibring, T; Baxandall, J; Harter, C C

    1984-02-01

    Extracellular sodium ion is required for activation of motility and respiration in sea urchin sperm when semen is diluted in seawater. We have investigated the role of sodium ion in maintenance of sperm activity. Active sperm lose activity on transfer to sodium-free artificial seawater and can be reactivated with external Na+. Reactivation occurs in the range of Na+ concentration required for initial activation; ammonium can substitute for sodium in reactivation. Sperm withdrawn from sodium and sperm prior to activation share a characteristic morphology with straight or gently bent flagella. Activation of sperm by amines in the absence of Na+ is unstable. It is followed by a steady respiratory decline which is temporarily reversed by addition of more amine and stably reversed by addition of Na+. Measurements of intracellular pH indicate that the internal pH rises during amine activation. Internal reacidification occurs during the period of respiratory decline, and Na+ again elevates internal pH. Treatment with cyanide abolishes the reacidification, indicating that it depends on respiration. We conclude that the sodium requirement persists in active sperm; respiration-dependent production of H+ must be balanced by sodium-dependent H+ removal to maintain activity.

  5. In Vitro Selection of Cathepsin E-Activity-Enhancing Peptide Aptamers at Neutral pH

    PubMed Central

    Biyani, Madhu; Futakami, Masae; Kitamura, Koichiro; Kawakubo, Tomoyo; Suzuki, Miho; Yamamoto, Kenji; Nishigaki, Koichi

    2011-01-01

    The aspartic protease cathepsin E has been shown to induce apoptosis in cancer cells under physiological conditions. Therefore, cathepsin E-activity-enhancing peptides functioning in the physiological pH range are valuable potential cancer therapeutic candidates. Here, we have used a general in vitro selection method (evolutionary rapid panning analysis system (eRAPANSY)), based on inverse substrate-function link (SF-link) selection to successfully identify cathepsin E-activity-enhancing peptide aptamers at neutral pH. A successive enrichment of peptide activators was attained in the course of selection. One such peptide activated cathepsin E up to 260%, had a high affinity (KD; ∼300 nM), and had physiological activity as demonstrated by its apoptosis-inducing reaction in cancerous cells. This method is expected to be widely applicable for the identification of protease-activity-enhancing peptide aptamers. PMID:21527983

  6. A neglected modulator of insulin-degrading enzyme activity and conformation: The pH.

    PubMed

    Grasso, Giuseppe; Satriano, Cristina; Milardi, Danilo

    2015-01-01

    Insulin-degrading enzyme (IDE), a ubiquitously expressed zinc metalloprotease, has multiple activities in addition to insulin degradation and its malfunction is believed to connect type 2 diabetes with Alzheimer's disease. IDE has been found in many different cellular compartments, where it may experience significant physio-pathological pH variations. However, the exact role of pH variations on the interplay between enzyme conformations, stability, oligomerization state and catalysis is not understood. Here, we use ESI mass spectrometry, atomic force microscopy, surface plasmon resonance and circular dichroism to investigate the structure-activity relationship of IDE at different pH values. We show that acidic pH affects the ability of the enzyme to bind the substrate and decrease the stability of the protein by inducing an α-helical bundle conformation with a concomitant dissociation of multi-subunit IDE assemblies into monomeric units and loss of activity. These effects suggest a major role played by electrostatic forces in regulating multi-subunit enzyme assembly and function. Our results clearly indicate a pH dependent coupling among enzyme conformation, assembly and stability and suggest that cellular acidosis can have a large effect on IDE oligomerization state, inducing an enzyme inactivation and an altered insulin degradation that could have an impact on insulin signaling.

  7. Use of nanoparticulate zinc oxide as intracanal medication in endodontics: pH and antimicrobial activity.

    PubMed

    Guerreiro-Tanomaru, Juliane M; Pereira, Kamila Figueiredo; Nascimento, Camila Almeida; Bernardi, Maria Inês Basso; Tanomaru-Filho, Mario

    2013-01-01

    The aim of this study was to evaluate the pH and antimicrobial activity of micro or nanoparticulate zinc oxide (ZnO) pastes with or without calcium hydroxide (CH). The following medications were evaluated: microparticulate ZnO + polyethylene glycol (PEG) 400; nanoparticulate ZnO + PEG 400; PEG 400; CH + microparticulate ZnO + PEG 400 and CH + nanoparticulate ZnO + PEG 400. The pH was assessed between 12 hours and 28 days, using a digital pH meter. The antimicrobial activity against Enterococcus faecalis (ATCC-9212), Candida albicans (ATCC-10231), Pseudomonas aeruginosa (ATCC-27853), Staphylococcus aureus (ATCC-6538) and Kocuria rhizophila (ATCC-9341) was determined in triplicate using agar diffusion test. The results were submitted to Kruskal-Wallis/Dunn and ANOVA/Tukey tests with 5% significance. The highest pH values were found for CH+ZnO, with higher values for nanoparticulate ZnO after 12 hours and 21 days (p < 0.05). CH+ZnO medication promoted higher growth inhibition against P. aeruginosa and lower against E. faecalis. Calcium hydroxide pastes have higher pH and antimicrobial activity when associated with either micro- or nanoparticulate zinc oxide.

  8. Effect of pH on the formation of lysosome-alginate beads for antimicrobial activity.

    PubMed

    Park, Hyun Jung; Min, Jiho; Ahn, Joo-Myung; Cho, Sung-Jin; Ahn, Ji-Young; Kim, Yang-Hoon

    2015-02-01

    In this study, we developed lysosome-alginate beads for application as an oral drug delivery system (ODDS). The beads harboring lysosomes, which have antimicrobial activity, and various concentrations of alginate were characterized and optimized. For application as an ODDS, pH-dependent lysosome-alginate beads were generated, and the level of lysosome release was investigated by using antimicrobial tests. At low pH, lysosomes were not released from the lysosome-alginate beads; however, at neutral pH, similar to the pH in the intestine, lysosome release was confirmed, as determined by a high antimicrobial activity. This study shows the potential of such an ODDS for the in vivo treatment of infection with pathogens.

  9. Influence of extracellular pH on growth, viability, cell size, acidification activity, and intracellular pH of Lactococcus lactis in batch fermentations.

    PubMed

    Hansen, Gunda; Johansen, Claus Lindvald; Marten, Gunvor; Wilmes, Jacqueline; Jespersen, Lene; Arneborg, Nils

    2016-07-01

    In this study, we investigated the influence of three extracellular pH (pHex) values (i.e., 5.5, 6.5, and 7.5) on the growth, viability, cell size, acidification activity in milk, and intracellular pH (pHi) of Lactococcus lactis subsp. lactis DGCC1212 during pH-controlled batch fermentations. A universal parameter (e.g., linked to pHi) for the description or prediction of viability, specific acidification activity, or growth behavior at a given pHex was not identified. We found viability as determined by flow cytometry to remain high during all growth phases and irrespectively of the pH set point. Furthermore, regardless of the pHex, the acidification activity per cell decreased over time which seemed to be linked to cell shrinkage. Flow cytometric pHi determination demonstrated an increase of the averaged pHi level for higher pH set points, while the pH gradient (pHi-pHex) and the extent of pHi heterogeneity decreased. Cells maintained positive pH gradients at a low pHex of 5.5 and even during substrate limitation at the more widely used pHex 6.5. Moreover, the strain proved able to grow despite small negative or even absent pH gradients at a high pHex of 7.5. The larger pHi heterogeneity at pHex 5.5 and 6.5 was associated with more stressful conditions resulting, e.g., from higher concentrations of non-dissociated lactic acid, while the low pHi heterogeneity at pHex 7.5 most probably corresponded to lower concentrations of non-dissociated lactic acid which facilitated the cells to reach the highest maximum active cell counts of the three pH set points.

  10. 47 CFR 27.1160 - Cost-sharing requirements for AWS.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... would occur. All AWS entities that benefit from the clearance of this spectrum by other AWS entities or... AWS licensee enters into a spectrum leasing arrangement (as set forth in part 1, subpart X of this chapter) and the spectrum lessee triggers a cost-sharing obligation, the licensee is the AWS...

  11. Improved volatile fatty acids anaerobic production from waste activated sludge by pH regulation: Alkaline or neutral pH?

    PubMed

    Ma, Huijun; Chen, Xingchun; Liu, He; Liu, Hongbo; Fu, Bo

    2016-02-01

    In this study, the anaerobic fermentation was carried out for volatile fatty acids (VFAs) production at different pH (between 7.0 and 10.0) conditions with untreated sludge and heat-alkaline pretreated waste activated sludge. In the fermentation with untreated sludge, the extent of hydrolysis of organic matters and extent of acidification at alkaline pH are 54.37% and 30.37%, respectively, resulting in the highest VFAs yield at 235.46mg COD/gVS of three pH conditions. In the fermentation with heat-alkaline pretreated sludge, the acidification rate and VFAs yield at neutral pH are 30.98% and 240.14mg COD/gVS, respectively, which are higher than that at other pH conditions. With the glucose or bovine serum albumin as substrate for VFAs production, the neutral pH showed a higher VFAs concentration than the alkaline pH condition. The results of terminal restriction fragment length polymorphism (T-RFLP) analysis indicated that the alkaline pH caused low microbial richness. Based on the results in this study, we demonstrated that the alkaline pH is favor of hydrolysis of organic matter in sludge while neutral pH improved the acidogenesis for the VFAs production from sludge. Our finding is obvious different to the previous research and helpful for the understanding of how heat-alkaline pretreatment and alkaline fermentation influence the VFAs production, and beneficial to the development of VFAs production process.

  12. An aposymbiotic primary coral polyp counteracts acidification by active pH regulation

    NASA Astrophysics Data System (ADS)

    Ohno, Yoshikazu; Iguchi, Akira; Shinzato, Chuya; Inoue, Mayuri; Suzuki, Atsushi; Sakai, Kazuhiko; Nakamura, Takashi

    2017-01-01

    Corals build their skeletons using extracellular calcifying fluid located in the tissue–skeleton interface. However, the mechanism by which corals control the transport of calcium and other ions from seawater and the mechanism of constant alkalization of calcifying fluid are largely unknown. To address these questions, we performed direct pH imaging at calcification sites (subcalicoblastic medium, SCM) to visualize active pH upregulation in live aposymbiotic primary coral polyps treated with HCl-acidified seawater. Active alkalization was observed in all individuals using vital staining method while the movement of HPTS and Alexa Fluor to SCM suggests that certain ions such as H+ could diffuse via a paracellular pathway to SCM. Among them, we discovered acid-induced oscillations in the pH of SCM (pHSCM), observed in 24% of polyps examined. In addition, we discovered acid-induced pH up-regulation waves in 21% of polyps examined, which propagated among SCMs after exposure to acidified seawater. Our results showed that corals can regulate pHSCM more dynamically than was previously believed. These observations will have important implications for determining how corals regulate pHSCM during calcification. We propose that corals can sense ambient seawater pH via their innate pH-sensitive systems and regulate pHSCM using several unknown pH-regulating ion transporters that coordinate with multicellular signaling occurring in coral tissue.

  13. An aposymbiotic primary coral polyp counteracts acidification by active pH regulation.

    PubMed

    Ohno, Yoshikazu; Iguchi, Akira; Shinzato, Chuya; Inoue, Mayuri; Suzuki, Atsushi; Sakai, Kazuhiko; Nakamura, Takashi

    2017-01-18

    Corals build their skeletons using extracellular calcifying fluid located in the tissue-skeleton interface. However, the mechanism by which corals control the transport of calcium and other ions from seawater and the mechanism of constant alkalization of calcifying fluid are largely unknown. To address these questions, we performed direct pH imaging at calcification sites (subcalicoblastic medium, SCM) to visualize active pH upregulation in live aposymbiotic primary coral polyps treated with HCl-acidified seawater. Active alkalization was observed in all individuals using vital staining method while the movement of HPTS and Alexa Fluor to SCM suggests that certain ions such as H(+) could diffuse via a paracellular pathway to SCM. Among them, we discovered acid-induced oscillations in the pH of SCM (pHSCM), observed in 24% of polyps examined. In addition, we discovered acid-induced pH up-regulation waves in 21% of polyps examined, which propagated among SCMs after exposure to acidified seawater. Our results showed that corals can regulate pHSCM more dynamically than was previously believed. These observations will have important implications for determining how corals regulate pHSCM during calcification. We propose that corals can sense ambient seawater pH via their innate pH-sensitive systems and regulate pHSCM using several unknown pH-regulating ion transporters that coordinate with multicellular signaling occurring in coral tissue.

  14. An aposymbiotic primary coral polyp counteracts acidification by active pH regulation

    PubMed Central

    Ohno, Yoshikazu; Iguchi, Akira; Shinzato, Chuya; Inoue, Mayuri; Suzuki, Atsushi; Sakai, Kazuhiko; Nakamura, Takashi

    2017-01-01

    Corals build their skeletons using extracellular calcifying fluid located in the tissue–skeleton interface. However, the mechanism by which corals control the transport of calcium and other ions from seawater and the mechanism of constant alkalization of calcifying fluid are largely unknown. To address these questions, we performed direct pH imaging at calcification sites (subcalicoblastic medium, SCM) to visualize active pH upregulation in live aposymbiotic primary coral polyps treated with HCl-acidified seawater. Active alkalization was observed in all individuals using vital staining method while the movement of HPTS and Alexa Fluor to SCM suggests that certain ions such as H+ could diffuse via a paracellular pathway to SCM. Among them, we discovered acid-induced oscillations in the pH of SCM (pHSCM), observed in 24% of polyps examined. In addition, we discovered acid-induced pH up-regulation waves in 21% of polyps examined, which propagated among SCMs after exposure to acidified seawater. Our results showed that corals can regulate pHSCM more dynamically than was previously believed. These observations will have important implications for determining how corals regulate pHSCM during calcification. We propose that corals can sense ambient seawater pH via their innate pH-sensitive systems and regulate pHSCM using several unknown pH-regulating ion transporters that coordinate with multicellular signaling occurring in coral tissue. PMID:28098180

  15. Illustrating the Effect of pH on Enzyme Activity Using Gibbs Energy Profiles

    ERIC Educational Resources Information Center

    Bearne, Stephen L.

    2014-01-01

    Gibbs energy profiles provide students with a visual representation of the energy changes that occur during enzyme catalysis, making such profiles useful as teaching and learning tools. Traditional kinetic topics, such as the effect of pH on enzyme activity, are often not discussed in terms of Gibbs energy profiles. Herein, the symbolism of Gibbs…

  16. Proteolytic Activity at Alkaline pH in Oat Leaves, Isolation of an Aminopeptidase 1

    PubMed Central

    Casano, Leonardo M.; Desimone, Marcelo; Trippi, Victorio S.

    1989-01-01

    Proteolytic activity in oat leaf extracts was measured with both azocasein and ribulose bisphosphate carboxylase (Rubisco) as substrates over a wide range of pH (3.0-9.2). With either azocasein or Rubisco activity peaks appeared at pH 4.8, 6.6, and 8.4. An aminopeptidase (AP) which hydrolyzes leucine-nitroanilide was partially purified. Purification consisted of a series of six steps which included ammonium sulfate precipitation, gel filtration, and two ionic exchange chromatographies. The enzyme was purified more than 100-fold. The apparent Km for leucine-nitroanilide is 0.08 millimolar at its pH optimum of 8.4. AP may be a cystein protease since it is inhibited by heavy metals and activated by 2-mercaptoethanol. Isolated chloroplasts were also able to hydrolyze leucine-nitroanilide at a pH optimum of 8.4, indicating that AP could be localized inside the photosynthetic organelles. PMID:16667194

  17. Enzymatic Characteristics of a Highly Thermostable β-(1-4)-Glucanase from Fervidobacterium islandicum AW-1 (KCTC 4680).

    PubMed

    Jeong, Woo Soo; Seo, Dong Ho; Jung, Jong Hyun; Jung, Dong Hyun; Lee, Dong-Woo; Park, Young-Seo; Park, Cheonseok

    2017-02-28

    A highly thermostable β-(1-4)-glucanase (NA23_08975) gene (fig) from Fervidobacterium islandicum AW-1, a native-feather degrading thermophilic eubacterium, was cloned and expressed in Escherichia coli. The recombinant FiG (rFiG) protein showed strong activity toward β-D-glucan from barley (367.0 IU/mg), galactomannan (174.0 IU/mg), and 4-nitrophenyl-cellobioside (66.1 IU/mg), but relatively weak activity was observed with hydroxyethyl cellulose (5.3 IU/mg), carboxymethyl cellulose (2.4 IU/mg), and xylan from oat spelt (1.4 IU/mg). rFiG exhibited optimal activity at 90°C and pH 5.0. In addition, this enzyme was extremely thermostable, showing a half-life of 113 h at 85°C. These results indicate that rFiG could be used for hydrolysis of cellulosic and hemicellulosic biomass substrates for biofuel production.

  18. pH control of the structure, composition, and catalytic activity of sulfated zirconia

    SciTech Connect

    Ivanov, Vladimir K.; Baranchikov, Alexander Ye.; Kopitsa, Gennady P.; Lermontov, Sergey A.; Yurkova, Lyudmila L.; Gubanova, Nadezhda N.; Ivanova, Olga S.; Lermontov, Anatoly S.; Rumyantseva, Marina N.; Vasilyeva, Larisa P.; Sharp, Melissa; Pranzas, P. Klaus; Tretyakov, Yuri D.

    2013-02-15

    We report a detailed study of structural and chemical transformations of amorphous hydrous zirconia into sulfated zirconia-based superacid catalysts. Precipitation pH is shown to be the key factor governing structure, composition and properties of amorphous sulfated zirconia gels and nanocrystalline sulfated zirconia. Increase in precipitation pH leads to substantial increase of surface fractal dimension (up to {approx}2.7) of amorphous sulfated zirconia gels, and consequently to increase in specific surface area (up to {approx}80 m{sup 2}/g) and simultaneously to decrease in sulfate content and total acidity of zirconia catalysts. Complete conversion of hexene-1 over as synthesized sulfated zirconia catalysts was observed even under ambient conditions. - Graphical abstract: Surface fractal dimension of amorphous sulfated zirconia and specific surface area and catalytic activity of crystalline sulfated zirconia as a function of precipitation pH. Highlights: Black-Right-Pointing-Pointer Structural transformation of amorphous hydrous zirconia into sulfated zirconia is studied. Black-Right-Pointing-Pointer Precipitation pH controls surface fractal dimension of amorphous zirconia gels. Black-Right-Pointing-Pointer Precipitation pH is the key factor governing properties of sulfated zirconia.

  19. Algal and bacterial activities in acidic (pH 3) strip mine lakes

    SciTech Connect

    Gyure, R.A.; Konopka, A.; Brooks, A.; Doemel, W.

    1987-09-01

    Reservoir 29 and Lake B are extremely acid lakes (epilimnion pHs of 2.7 and 3.2, respectively), because they receive acidic discharges from coal refuse piles. They differ in that the pH of profundal sediments in Reservoir 29 increased from 2.7 to 3.8 during the period of thermal stratification, whereas permanently anoxic sediments in Lake B had a pH of 6.2. The pH rise in Reservoir 29 sediments was correlated with a temporal increase in H/sub 2/S concentration in the anaerobic hypolimnion from 0 to >1 mM. The chlorophyll a levels in the epilimnion of Reservoir 29 were low, and the rate of primary production was typical of an oligotrophic system. However, there was a dense 10-cm layer of algal biomass at the bottom of the metalimnion. Production by this layer was low owing to light limitation and possibly H/sub 2/S toxicity. The specific photosynthetic rates of epilimnetic algae were low, which suggests that nutrient availability is more important than pH in limiting production. The highest photosynthetic rates were obtained in water samples incubated at pH 2.7 to 4. Heterotrophic bacterial activity (measured by (/sup 14/C)glucose metabolism) was greatest at the sediment/water interface. Bacterial production (assayed by thymidine incorporation) was as high in Reservoir 29 as in a nonacid mesotrophic Indiana lake.

  20. Algal and Bacterial Activities in Acidic (pH 3) Strip Mine Lakes

    PubMed Central

    Gyure, Ruth A.; Konopka, Allan; Brooks, Austin; Doemel, William

    1987-01-01

    Reservoir 29 and Lake B are extremely acid lakes (epilimnion pHs of 2.7 and 3.2, respectively), because they receive acidic discharges from coal refuse piles. They differ in that the pH of profundal sediments in Reservoir 29 increased from 2.7 to 3.8 during the period of thermal stratification, whereas permanently anoxic sediments in Lake B had a pH of 6.2. The pH rise in Reservoir 29 sediments was correlated with a temporal increase in H2S concentration in the anaerobic hypolimnion from 0 to >1 mM. The chlorophyll a levels in the epilimnion of Reservoir 29 were low, and the rate of primary production was typical of an oligotrophic system. However, there was a dense 10-cm layer of algal biomass at the bottom of the metalimnion. Production by this layer was low owing to light limitation and possibly H2S toxicity. The specific photosynthetic rates of epilimnetic algae were low, which suggests that nutrient availability is more important than pH in limiting production. The highest photosynthetic rates were obtained in water samples incubated at pH 2.7 to 4. Heterotrophic bacterial activity (measured by [14C]glucose metabolism) was greatest at the sediment/water interface. Bacterial production (assayed by thymidine incorporation) was as high in Reservoir 29 as in a nonacid mesotrophic Indiana lake. PMID:16347430

  1. Survival of Salmonella enterica serotype Tennessee during simulated gastric passage is improved by low water activity and high fat content.

    PubMed

    Aviles, Bryan; Klotz, Courtney; Smith, Twyla; Williams, Robert; Ponder, Monica

    2013-02-01

    The low water activity (a(w) 0.3) of peanut butter prohibits the growth of Salmonella in a product; however, illnesses are reported from peanut butter contaminated with very small doses, suggesting the food matrix itself influences the infectious dose of Salmonella, potentially by improving Salmonella's survival in the gastrointestinal tract. The purpose of our study was to quantify the survival of a peanut butter outbreak-associated strain of Salmonella enterica serotype Tennessee when inoculated into peanut butters with different fat contents and a(w) (high fat, high a(w); high fat, low a(w); low fat, high a(w); low fat, low a(w)) and then challenged with a simulated gastrointestinal system. Exposures to increased fat content and decreased a(w) both were associated with a protective effect on the survival of Salmonella Tennessee in the simulated gastric fluid compared with control cells. After a simulated intestinal phase, the populations of Salmonella Tennessee in the control and low-fat formulations were not significantly different; however, a 2-log CFU/g increase occurred in high-fat formulations. This study demonstrates that cross-protection from low-a(w) stress and the presence of high fat results in improved survival in the low pH of the stomach. The potential for interaction of food matrix and stress adaptations could influence the virulence of Salmonella and should be considered for risk analysis.

  2. Effect of medium, pH, and inoculum size on activity of ceftizoxime and Sch-34343 against anaerobic bacteria.

    PubMed Central

    Borobio, M V; Pascual, A; Dominguez, M C; Perea, E J

    1986-01-01

    The effect of two media, three pH values, and three inoculum densities on the activity of ceftizoxime and Sch-34343 against anaerobic bacteria was evaluated. The activities of both antimicrobial agents were affected by medium composition, especially against Bacteroides fragilis. Changes in pH and inoculum size affected only the activity of ceftizoxime against anaerobic bacteria. PMID:3466569

  3. Characterization of an invertase with pH tolerance and truncation of its N-terminal to shift optimum activity toward neutral pH.

    PubMed

    Du, Liqin; Pang, Hao; Wang, Zilong; Lu, Jian; Wei, Yutuo; Huang, Ribo

    2013-01-01

    Most invertases identified to date have optimal activity at acidic pH, and are intolerant to neutral or alkaline environments. Here, an acid invertase named uninv2 is described. Uninv2 contained 586 amino acids, with a 100 amino acids N-terminal domain, a catalytic domain and a C-terminal domain. With sucrose as the substrate, uninv2 activity was optimal at pH 4.5 and at 45°C. Removal of N-terminal domain of uninv2 has shifted the optimum pH to 6.0 while retaining its optimum temperaure at 45°C. Both uninv2 and the truncated enzyme retained highly stable at neutral pH at 37°C, and they were stable at their optimum pH at 4°C for as long as 30 days. These characteristics make them far superior to invertase from Saccharomyces cerevisiae, which is mostly used as industrial enzyme.

  4. [Phosphatase activity in Amoeba proteus at pH 9.0].

    PubMed

    Sopina, V A

    2007-01-01

    In the free-living amoeba Amoeba proteus (strain B), after PAAG disk-electrophoresis of the homogenate supernatant, at using 1-naphthyl phosphate as a substrate and pH 9.0, three forms of phosphatase activity were revealed; they were arbitrarily called "fast", "intermediate", and "slow" phosphatases. The fast phosphatase has been established to be a fraction of lysosomal acid phosphatase that preserves some low activity at alkaline pH. The question as to which particular class the intermediate phosphatase belongs to has remained unanswered: it can be both acid phosphatase and protein tyrosine phosphatase (PTP). Based on data of inhibitor analysis, large substrate specificity, results of experiments with reactivation by Zn ions after inactivation with EDTA, other than in the fast and intermediate phosphatases localization in the amoeba cell, it is concluded that only slow phosphatase can be classified as alkaline phosphatase (EC 3.1.3.1).

  5. [Doctor Bronisław Hager (1890-1969)--doctor and politician].

    PubMed

    Drozdz, Łukasz M; Iwaniszyn-Popowicz, Iwona J; Mokrus, Oliwia M; Madry, Łukasz J

    2008-01-01

    Doctor Bronisław Hager was born into the family of the Polish intelligentsia. He was brought up according to the Polish tradition with respect for freedom and independence. His father, Maxymilian was a fervent fighter in the struggle for independence of Silesia. His mother was involved in community service. Bronisław Hager finished his medical studies at Collegium Medicum of Jagiellonian University on 27th March 1914. Between World War I and World War II he actively fought for joining Silesia to Poland. During World War II he emigrated to London where he became one of members of the Polish government in exile. At the end of World War II he was a doctor of United Nations in Bavaria. He came back to the country on 22th November 1946. He settled in Tarnowskie Góry and worked at the community health center till 1967. He translated Sir Alexander Fleming's work on the discovery and using of penicillin. He died on 30th June 1969. Bronisław Hager was a good doctor, a great politician, a wonderful speaker and a real Pole.

  6. A shift in the optimum pH of phospholipase D produced by activating long-chain anions

    PubMed Central

    Quarles, R. H.; Dawson, R. M. C.

    1969-01-01

    1. The activity of phospholipase D (phosphatidylcholine phosphatidohydrolase, EC 3.1.4.4) towards ultrasonically treated phosphatidylcholine or large phosphatidylcholine particles activated with ether was maximal near pH5, and there was little activity above pH6. 2. When the enzyme was activated by the addition of phosphatidic acid to large phosphatidylcholine particles the pH optimum was shifted to pH6·5 irrespective of the amount of activator added. 3. When the enzyme was activated with low concentrations of dodecyl sulphate the pH optimum was 5·5 with little activity above pH6. With higher concentrations of dodecyl sulphate the pH–activity profile was shifted upwards towards a pH optimum of 6·5–6·6, the magnitude of the shift depending on the extent of the hydrolysis. 4. The shifts in the pH–activity profiles cannot be correlated with changes in the `surface pH' of the substrate particles calculated from the measurement of their ζ-potentials (electrophoretic mobilities). PMID:5821735

  7. The oxidation state of active site thiols determines activity of saccharopine dehydrogenase at low pH.

    PubMed

    Bobyk, Kostyantyn D; Kim, Sang Gon; Kumar, Vidya Prasanna; Kim, Sung-Kun; West, Ann H; Cook, Paul F

    2011-09-15

    Saccharopine dehydrogenase catalyzes the NAD-dependent conversion of saccharopine to generate L-lysine and α-ketoglutarate. A disulfide bond between cysteine 205 and cysteine 249, in the vicinity of the dinucleotide-binding site, is observed in structures of the apoenzyme, while a dithiol is observed in a structure with AMP bound, suggesting preferential binding of the dinucleotide to reduced enzyme. Mutation of C205 to S gave increased values of V/E(t) and V/KE(t) at pH 7 compared to wild type. Primary deuterium and solvent deuterium kinetic isotope effects suggest the catalytic pathway, which includes the hydride transfer and hydrolysis steps, contributes more to rate limitation in C205S, but the rates of the two steps relative to one another remain the same. There is a large increase in the rate constants V₁/E(t) and V₁/K(NAD)Et at pH values below 7 compared to WT. Data indicate the low pH increase in activity results from a decreased sensitivity of the C205S mutant enzyme to the protonation state of an enzyme group with a pK(a) of about 7, likely responsible for a pH-dependent conformational change. Reduction of WT and C205S mutant enzymes with TCEP gives equal activities at pH 6, consistent with the increased activity observed for the C205S mutant enzyme.

  8. Optical spectroscopy of SNHiTS15aw

    NASA Astrophysics Data System (ADS)

    Pignata, G.; Olivares, F.; Forster, F.; Smith, C.; Vivas, K.; Hamuy, M.; Martin, J. San; Maureira, J. C.; Cabrera, G.; Anderson, J.; Gonzalez-Gaitan, S.; Galbany, L.; Bufano, F.; de Jaeger, T.; Hsiao, E.; Munoz, R.; Vera, E.

    2015-03-01

    We report optical wavelength spectroscopy obtained using the Goodman instrument mounted on SOAR at CTIO on UT 2015-03-12.1, for SNHiTS15aw discovered by HiTS, the High Cadence Transient Survey (see ATELs #5949, #5956, #7099, #7108, #7115, #7122, #7131, #7132, #7146, #7148, #7221).

  9. Professor Bolesław Rutkowski's 70th birthday Laudatio

    PubMed Central

    Ostrowski, Janusz; Durlik, Magdalena; Dębska-Ślizień, Alicja; Klinger, Marian; Manitius, Jacek; Sułowicz, Władysław; Więcek, Andrzej; Zdrojewski, Zbigniew

    2015-01-01

    On 5 December 2014 Professor Bolesław Rutkowski, the outstanding Polish nephrologist, doctor and researcher, renowned organizer, our teacher, colleague and friend, will celebrate his 70th birthday. We would like to take this opportunity to present to you his life and his scientific achievements.

  10. Antibiotic Bactericidal Activity Is Countered by Maintaining pH Homeostasis in Mycobacterium smegmatis

    PubMed Central

    Bartek, I. L.; Reichlen, M. J.; Honaker, R. W.; Leistikow, R. L.; Clambey, E. T.; Scobey, M. S.; Hinds, A. B.; Born, S. E.; Covey, C. R.; Schurr, M. J.; Lenaerts, A. J.

    2016-01-01

    ABSTRACT Antibiotics target specific biosynthetic processes essential for bacterial growth. It is intriguing that several commonalities connect the bactericidal activity of seemingly disparate antibiotics, such as the numerous conditions that confer broad-spectrum antibiotic tolerance. Whether antibiotics kill in a manner unique to their specific targets or by a universal mechanism is a critical and contested subject. Herein, we demonstrate that the bactericidal activity of diverse antibiotics against Mycobacterium smegmatis and four evolutionarily divergent bacterial pathogens was blocked by conditions that worked to maintain intracellular pH homeostasis. Single-cell pH analysis demonstrated that antibiotics increased the cytosolic pH of M. smegmatis, while conditions that promoted proton entry into the cytosol prevented intracellular alkalization and antibiotic killing. These findings led to a hypothesis that posits antibiotic lethality occurs when antibiotics obstruct ATP-consuming biosynthetic processes while metabolically driven proton efflux is sustained despite the loss of proton influx via ATP synthase. Consequently, without a concomitant reduction in respiratory proton efflux, cell death occurs due to intracellular alkalization. Our findings indicate the effects of antibiotics on pH homeostasis should be considered a potential mechanism contributing to antibiotic lethality. IMPORTANCE Since the discovery of antibiotics, mortality due to bacterial infection has decreased dramatically. However, infections from difficult to treat bacteria such as Mycobacterium tuberculosis and multidrug-resistant pathogens have been on the rise. An understanding of the cascade of events that leads to cell death downstream of specific drug-target interactions is not well understood. We have discovered that killing by several classes of antibiotics was stopped by maintaining pH balance within the bacterial cell, consistent with a shared mechanism of antibiotic killing. Our

  11. Influence of radiopacifying agents on the solubility, pH and antimicrobial activity of portland cement.

    PubMed

    Weckwerth, Paulo Henrique; Machado, Adriano Cosme de Oliveira; Kuga, Milton Carlos; Vivan, Rodrigo Ricci; Polleto, Raquel da Silva; Duarte, Marco Antonio Hungaro

    2012-01-01

    The aim of this study was to evaluate the interference of the radiopacifiers bismuth oxide (BO), bismuth carbonate (BC), bismuth subnitrate (BS), and zirconiun oxide (ZO) on the solubility, alkalinity and antimicrobial properties of white Portland cement (WPC). The substances were incorporated to PC, at a ratio of 1:4 (v/v) and subjected to a solubility test. To evaluate the pH, the cements were inserted into retrograde cavities prepared in simulated acrylic teeth and immediately immersed in deionized water. The pH of the solution was measured at 3, 24, 72 and 168 h. The antimicrobial activity was evaluated by a radial diffusion method against the microorganisms S. aureus (ATCC 25923), P. aeruginosa (ATCC 27853), E. faecalis (ATCC 29212) and C. albicans (ATCC 10231). The zone of microbial growth inhibition was measured after 24 h. The addition of BS and BC increased the solubility of the cement. The pH values demonstrated that all materials produced alkaline levels. At 3 h, BS showed lower pH than WPC (p<0.05). At 168 h, all materials showed similar pHs (p>0.05). The materials did not present antimicrobial activity for S. aureus, P. aeruginosas and E. faecalis (p>0.05). With regards to C. albicans, all materials formed an inhibition zone, mainly the mixture of WPC with ZO (p<0.05). The type of radiopacifier incorporated into WPC interfered with its physical and antimicrobial properties. ZO was found to be a viable radiopacifier that can be used with WPC.

  12. Acceptance/operational test procedure 101-AW tank camera purge system and 101-AW video camera system

    SciTech Connect

    Castleberry, J.L.

    1994-09-19

    This procedure will document the satisfactory operation of the 101-AW Tank Camera Purge System (CPS) and the 101-AW Video Camera System. The safety interlock which shuts down all the electronics inside the 101-AW vapor space, during loss of purge pressure, will be in place and tested to ensure reliable performance. This procedure is separated into four sections. Section 6.1 is performed in the 306 building prior to delivery to the 200 East Tank Farms and involves leak checking all fittings on the 101-AW Purge Panel for leakage using a Snoop solution and resolving the leakage. Section 7.1 verifies that PR-1, the regulator which maintains a positive pressure within the volume (cameras and pneumatic lines), is properly set. In addition the green light (PRESSURIZED) (located on the Purge Control Panel) is verified to turn on above 10 in. w.g. and after the time delay (TDR) has timed out. Section 7.2 verifies that the purge cycle functions properly, the red light (PURGE ON) comes on, and that the correct flowrate is obtained to meet the requirements of the National Fire Protection Association. Section 7.3 verifies that the pan and tilt, camera, associated controls and components operate correctly. This section also verifies that the safety interlock system operates correctly during loss of purge pressure. During the loss of purge operation the illumination of the amber light (PURGE FAILED) will be verified.

  13. Inorganic and Radiochemical Analysis of AW-101 and AN-107 Tank Waste

    SciTech Connect

    MW Urie; JJ Wagner; LR Greenwood; OT Farmer; SK Fiskum; RT Ratner; CZ Soderquist

    1999-11-11

    This report presents the inorganic and radiochemical analytical results for AW-101 and AN-107 as received materials. The analyses were conducted in support of the BNFL Proposal No. 30406/29274 Task 5.0. The inorganic and radiochemical analysis results obtained from the as received materials are used to provide initial characterization information for subsequent process testing and to provide data to support permit application activities. Quality Assurance (QA) Plan MCS-033 provides the operational and quality control protocols for the analytical activities, and whenever possible, analyses were performed to SW-846 equivalent methods and protocols.

  14. Intracellular pH regulates basolateral K+ and Cl- conductances in colonic epithelial cells by modulating Ca2+ activation

    PubMed Central

    1991-01-01

    The role of intracellular pH as a modulator of basolateral K+ and Cl- conductances in epithelial cells was studied using digitonin- permeabilized colonic cell layers so that cytosolic pH could be clamped at specific values, while basolateral K+ and Cl- conductances were activated by stepwise increases in intracellular free Ca2+. Increasing the intracellular pH from 6.6 to 8.0 enhanced the sensitivity of both ionic conductances to intracellular Ca2+, but changing extracellular pH had no effect. Maximal K+ and Cl- currents activated by Ca2+ were not affected by changes in intracellular pH, suggesting that protons do not alter the conduction properties of the channels. Hill analysis of the Ca2+ activation process revealed that raising the cytosolic pH from 6.6 to 8.0 reduced the K1/2 for Ca2+ activation. In the absence of Ca2+, changes in intracellular pH did not have a significant effect on the basolateral K+ and Cl- conductances. These results are consistent with the notion that changes in cytosolic pH can modulate basolateral conductances by modifying the action of calcium, perhaps by acting at or near the activation site to provide a mechanism of variable "gain control." PMID:1719125

  15. Regulation of the voltage-insensitive step of HERG activation by extracellular pH.

    PubMed

    Zhou, Qinlian; Bett, Glenna C L

    2010-06-01

    Human ether-à-go-go-related gene (HERG, Kv11.1, KCNH2) voltage-gated K(+) channels dominate cardiac action potential repolarization. In addition, HERG channels play a role in neuronal and smooth cell excitability as well as cancer pathology. Extracellular pH (pH(o)) is modified during myocardial ischemia, inflammation, and respiratory alkalosis, so understanding the response of HERG channels to changes in pH is of clinical significance. The relationship between pH(o) and HERG channel gating appears complex. Acidification has previously been reported to speed, slow, or have no effect on activation. We therefore undertook comprehensive analysis of the effect of pH(o) on HERG channel activation. HERG channels have unique and complex activation gating characteristics with both voltage-sensitive and voltage-insensitive steps in the activation pathway. Acidosis decreased the activation rate, suppressed peak current, and altered the sigmoidicity of gating near threshold potentials. At positive voltages, where the voltage-insensitive transition is rate limiting, pH(o) modified the voltage-insensitive step with a pK(a) similar to that of histidine. Hill coefficient analysis was incompatible with a coefficient of 1 but was well described by a Hill coefficient of 4. We derived a pH(o)-sensitive term for a five-state Markov model of HERG channel gating. This model demonstrates the mechanism of pH(o) sensitivity in HERG channel activation. Our experimental data and mathematical model demonstrate that the pH(o) sensitivity of HERG channel activation is dominated by the pH(o) sensitivity of the voltage-insensitive step, in a fashion that is compatible with the presence of at least one proton-binding site on each subunit of the channel tetramer.

  16. 76 FR 11529 - Gregory Desobry, Ph.D.; Order Requiring Notification of Involvement in NRC-Licensed Activities

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-02

    ... From the Federal Register Online via the Government Publishing Office NUCLEAR REGULATORY COMMISSION Gregory Desobry, Ph.D.; Order Requiring Notification of Involvement in NRC-Licensed Activities I... Mr. Desobry, Ph.D., requests a hearing, that person shall set forth with particularity the manner...

  17. Genomic and Secondary Metabolite Analyses of Streptomyces sp. 2AW Provide Insight into the Evolution of the Cycloheximide Pathway

    PubMed Central

    Stulberg, Elizabeth R.; Lozano, Gabriel L.; Morin, Jesse B.; Park, Hyunjun; Baraban, Ezra G.; Mlot, Christine; Heffelfinger, Christopher; Phillips, Gillian M.; Rush, Jason S.; Phillips, Andrew J.; Broderick, Nichole A.; Thomas, Michael G.; Stabb, Eric V.; Handelsman, Jo

    2016-01-01

    The dearth of new antibiotics in the face of widespread antimicrobial resistance makes developing innovative strategies for discovering new antibiotics critical for the future management of infectious disease. Understanding the genetics and evolution of antibiotic producers will help guide the discovery and bioengineering of novel antibiotics. We discovered an isolate in Alaskan boreal forest soil that had broad antimicrobial activity. We elucidated the corresponding antimicrobial natural products and sequenced the genome of this isolate, designated Streptomyces sp. 2AW. This strain illustrates the chemical virtuosity typical of the Streptomyces genus, producing cycloheximide as well as two other biosynthetically unrelated antibiotics, neutramycin, and hygromycin A. Combining bioinformatic and chemical analyses, we identified the gene clusters responsible for antibiotic production. Interestingly, 2AW appears dissimilar from other cycloheximide producers in that the gene encoding the polyketide synthase resides on a separate part of the chromosome from the genes responsible for tailoring cycloheximide-specific modifications. This gene arrangement and our phylogenetic analyses of the gene products suggest that 2AW holds an evolutionarily ancestral lineage of the cycloheximide pathway. Our analyses support the hypothesis that the 2AW glutaramide gene cluster is basal to the lineage wherein cycloheximide production diverged from other glutarimide antibiotics. This study illustrates the power of combining modern biochemical and genomic analyses to gain insight into the evolution of antibiotic-producing microorganisms. PMID:27199910

  18. Genomic and Secondary Metabolite Analyses of Streptomyces sp. 2AW Provide Insight into the Evolution of the Cycloheximide Pathway.

    PubMed

    Stulberg, Elizabeth R; Lozano, Gabriel L; Morin, Jesse B; Park, Hyunjun; Baraban, Ezra G; Mlot, Christine; Heffelfinger, Christopher; Phillips, Gillian M; Rush, Jason S; Phillips, Andrew J; Broderick, Nichole A; Thomas, Michael G; Stabb, Eric V; Handelsman, Jo

    2016-01-01

    The dearth of new antibiotics in the face of widespread antimicrobial resistance makes developing innovative strategies for discovering new antibiotics critical for the future management of infectious disease. Understanding the genetics and evolution of antibiotic producers will help guide the discovery and bioengineering of novel antibiotics. We discovered an isolate in Alaskan boreal forest soil that had broad antimicrobial activity. We elucidated the corresponding antimicrobial natural products and sequenced the genome of this isolate, designated Streptomyces sp. 2AW. This strain illustrates the chemical virtuosity typical of the Streptomyces genus, producing cycloheximide as well as two other biosynthetically unrelated antibiotics, neutramycin, and hygromycin A. Combining bioinformatic and chemical analyses, we identified the gene clusters responsible for antibiotic production. Interestingly, 2AW appears dissimilar from other cycloheximide producers in that the gene encoding the polyketide synthase resides on a separate part of the chromosome from the genes responsible for tailoring cycloheximide-specific modifications. This gene arrangement and our phylogenetic analyses of the gene products suggest that 2AW holds an evolutionarily ancestral lineage of the cycloheximide pathway. Our analyses support the hypothesis that the 2AW glutaramide gene cluster is basal to the lineage wherein cycloheximide production diverged from other glutarimide antibiotics. This study illustrates the power of combining modern biochemical and genomic analyses to gain insight into the evolution of antibiotic-producing microorganisms.

  19. Active Control of pH in the Bioculture System Through Carbon Dioxide Control

    NASA Technical Reports Server (NTRS)

    Monhollon, Luke; Pletcher, David; Hauss, Jessica

    2016-01-01

    For successful cell research, the growth culture environment must be tightly controlled. Deviance from the optimal conditions will mask the desired variable being analyzed or lead to inconstancies in the results. In standard laboratories, technology and procedures are readily available for the reliable control of variables such as temperature, pH, nutrient loading, and dissolved gases. Due to the nature of spaceflight, and the inherent constraints to engineering designs, these same elements become a challenge to maintain at stable values by both automated and manual approaches. Launch mass, volume, and power usage create significant constraints to cell culture systems; nonetheless, innovative solutions for active environmental controls are available. The acidity of the growth media cannot be measured through standard probes due to the degradation of electrodes and reliance on indicators for chromatography. Alternatively, carbon dioxide sensors are capable of monitoring the pH by leveraging the relationship between the partial pressure of carbon dioxide and carbonic acid in solution across a membrane. In microgravity cell growth systems, the gas delivery system can be used to actively maintain the media at the proper acidity by maintaining a suitable gas mixture around permeable tubing. Through this method, launch mass and volume are significantly reduced through the efficient use of the limited gas supply in orbit.

  20. The influence of soil pH on the diversity, abundance and transcriptional activity of ammonia oxidizing archaea and bacteria.

    PubMed

    Nicol, Graeme W; Leininger, Sven; Schleper, Christa; Prosser, James I

    2008-11-01

    Autotrophic ammonia oxidation occurs in acid soils, even though laboratory cultures of isolated ammonia oxidizing bacteria fail to grow below neutral pH. To investigate whether archaea possessing ammonia monooxygenase genes were responsible for autotrophic nitrification in acid soils, the community structure and phylogeny of ammonia oxidizing bacteria and archaea were determined across a soil pH gradient (4.9-7.5) by amplifying 16S rRNA and amoA genes followed by denaturing gradient gel electrophoresis (DGGE) and sequence analysis. The structure of both communities changed with soil pH, with distinct populations in acid and neutral soils. Phylogenetic reconstructions of crenarchaeal 16S rRNA and amoA genes confirmed selection of distinct lineages within the pH gradient and high similarity in phylogenies indicated a high level of congruence between 16S rRNA and amoA genes. The abundance of archaeal and bacterial amoA gene copies and mRNA transcripts contrasted across the pH gradient. Archaeal amoA gene and transcript abundance decreased with increasing soil pH, while bacterial amoA gene abundance was generally lower and transcripts increased with increasing pH. Short-term activity was investigated by DGGE analysis of gene transcripts in microcosms containing acidic or neutral soil or mixed soil with pH readjusted to that of native soils. Although mixed soil microcosms contained identical archaeal ammonia oxidizer communities, those adapted to acidic or neutral pH ranges showed greater relative activity at their native soil pH. Findings indicate that different bacterial and archaeal ammonia oxidizer phylotypes are selected in soils of different pH and that these differences in community structure and abundances are reflected in different contributions to ammonia oxidizer activity. They also suggest that both groups of ammonia oxidizers have distinct physiological characteristics and ecological niches, with consequences for nitrification in acid soils.

  1. The voltage-activated hydrogen ion conductance in rat alveolar epithelial cells is determined by the pH gradient

    PubMed Central

    1995-01-01

    Voltage-activated H+ currents were studied in rat alveolar epithelial cells using tight-seal whole-cell voltage clamp recording and highly buffered, EGTA-containing solutions. Under these conditions, the tail current reversal potential, Vrev, was close to the Nernst potential, EH, varying 52 mV/U pH over four delta pH units (delta pH = pHo - pHi). This result indicates that H+ channels are extremely selective, PH/PTMA > 10(7), and that both internal and external pH, pHi, and pHo, were well controlled. The H+ current amplitude was practically constant at any fixed delta pH, in spite of up to 100-fold symmetrical changes in H+ concentration. Thus, the rate-limiting step in H+ permeation is pH independent, must be localized to the channel (entry, permeation, or exit), and is not bulk diffusion limitation. The instantaneous current- voltage relationship exhibited distinct outward rectification at symmetrical pH, suggesting asymmetry in the permeation pathway. Sigmoid activation kinetics and biexponential decay of tail currents near threshold potentials indicate that H+ channels pass through at least two closed states before opening. The steady state H+ conductance, gH, as well as activation and deactivation kinetic parameters were all shifted along the voltage axis by approximately 40 mV/U pH by changes in pHi or pHo, with the exception of the fast component of tail currents which was shifted less if at all. The threshold potential at which H+ currents were detectably activated can be described empirically as approximately 20-40(pHo-pHi) mV. If internal and external protons regulate the voltage dependence of gH gating at separate sites, then they must be equally effective. A simpler interpretation is that gating is controlled by the pH gradient, delta pH. We propose a simple general model to account for the observed delta pH dependence. Protonation at an externally accessible site stabilizes the closed channel conformation. Deprotonation of this site permits a

  2. Species composition and seasonal dynamics of mosquito larvae in the Wrocław, Poland area.

    PubMed

    Rydzanicz, K; Lonc, E

    2003-12-01

    A total of 13,214 larvae specimens representing eight species (Culex pipiens, Culiseta annulata, Ochlerotatus cantans, Ochlerotatus communis, Ochlerotatus excrucians, Ochlerotatus sticticus, Aedes vexans, and Anopheles maculipennis) from twelve sampling places in the Wrocław area in Poland were collected between May and October, 1998-2000. The development rate of each mosquito species depended on hydrological types and various plant communities in water bodies. The water quality (temperature, pH and ion concentration) influenced mosquito larvae dynamics as well. The most abundant and constant species was Cx. pipiens. Its monthly population was significantly higher in the water bodies located in the city center as compared to those in the suburbs.

  3. Variations of thiaminase I activity pH dependencies among typical Great Lakes forage fish and Paenibacillus thiaminolyticus.

    USGS Publications Warehouse

    Zajicek, J.L.; Brown, L.; Brown, S.B.; Honeyfield, D.C.; Fitzsimons, J.D.; Tillitt, D.E.

    2009-01-01

    The source of thiaminase in the Great Lakes food web remains unknown. Biochemical characterization of the thiaminase I activities observed in forage fish was undertaken to provide insights into potential thiaminase sources and to optimize catalytic assay conditions. We measured the thiaminase I activities of crude extracts from five forage fish species and one strain of Paenibacillus thiaminolyticus over a range of pH values. The clupeids, alewife Alosa pseudoharengus and gizzard shad Dorosoma cepedianum, had very similar thiaminase I pH dependencies, with optimal activity ranges (> or = 90% of maximum activity) between pH 4.6 and 5.5. Rainbow smelt Osmerus mordax and spottail shiner Notropis hudsonius had optimal activity ranges between pH 5.5-6.6. The thiaminase I activity pH dependence profile of P. thiaminolyticus had an optimal activity range between pH 5.4 and 6.3, which was similar to the optimal range for rainbow smelt and spottail shiners. Incubation of P. thiaminolyticus extracts with extracts from bloater Coregonus hoyi (normally, bloaters have little or no detectable thiaminase I activity) did not significantly alter the pH dependence profile of P. thiaminolyticus-derived thiaminase I, such that it continued to resemble that of the rainbow smelt and spottail shiner, with an apparent optimal activity range between pH 5.7 and 6.6. These data are consistent with the hypothesis of a bacterial source for thiaminase I in the nonclupeid species of forage fish; however, the data also suggest different sources of thiaminase I enzymes in the clupeid species.

  4. Voltage dependence and pH regulation of human polycystin-2-mediated cation channel activity.

    PubMed

    Gonzalez-Perrett, Silvia; Batelli, Marisa; Kim, Keetae; Essafi, Makram; Timpanaro, Gustavo; Moltabetti, Nicolas; Reisin, Ignacio L; Arnaout, M Amin; Cantiello, Horacio F

    2002-07-12

    Polycystin-2, the product of the human PKD2 gene, whose mutations cause autosomal dominant polycystic kidney disease, is a large conductance, Ca(2+)-permeable non-selective cation channel. Polycystin-2 is functionally expressed in the apical membrane of the human syncytiotrophoblast, where it may play a role in the control of fetal electrolyte homeostasis. Little is known, however, about the mechanisms that regulate polycystin-2 channel function. In this study, the role of pH in the regulation of polycystin-2 was assessed by ion channel reconstitution of both apical membranes of human syncytiotrophoblast and the purified FLAG-tagged protein from in vitro transcribed/translated material. A kinetic analysis of single channel currents, including dwell time histograms, confirmed two open and two close states for spontaneous channel behavior and a strong voltage dependence of the open probability of the channel (P(o)). A reduction of cis pH (pH(cis)) decreased P(o) and shifted the voltage dependence of channel function but had no effect on the single channel conductance. An increase in pH(cis), in contrast, increased NP(o) (channel number times P(o)). Elimination of the H(+) chemical gradient did not reverse the low pH(cis) inhibition of polycystin-2. Similar findings confirmed the pH effect on the in vitro translated, FLAG-tagged purified polycystin-2. The data indicate the presence of an H(+) ion regulatory site in the channel protein, which is accessible from the cytoplasmic side of the protein. This protonation site controls polycystin-2 cation-selective channel activity.

  5. An enzyme kinetics study of the pH dependence of chloride activation of oxygen evolution in photosystem II.

    PubMed

    Baranov, Sergei; Haddy, Alice

    2017-03-01

    Oxygen evolution by photosystem II (PSII) involves activation by Cl(-) ion, which is regulated by extrinsic subunits PsbQ and PsbP. In this study, the kinetics of chloride activation of oxygen evolution was studied in preparations of PSII depleted of the PsbQ and PsbP subunits (NaCl-washed and Na2SO4/pH 7.5-treated) over a pH range from 5.3 to 8.0. At low pH, activation by chloride was followed by inhibition at chloride concentrations >100 mM, whereas at high pH activation continued as the chloride concentration increased above 100 mM. Both activation and inhibition were more pronounced at lower pH, indicating that Cl(-) binding depended on protonation events in each case. The simplest kinetic model that could account for the complete data set included binding of Cl(-) at two sites, one for activation and one for inhibition, and four protonation steps. The intrinsic (pH-independent) dissociation constant for Cl(-) activation, K S, was found to be 0.9 ± 0.2 mM for both preparations, and three of the four pK as were determined, with the fourth falling below the pH range studied. The intrinsic inhibition constant, K I, was found to be 64 ± 2 and 103 ± 7 mM for the NaCl-washed and Na2SO4/pH7.5-treated preparations, respectively, and is considered in terms of the conditions likely to be present in the thylakoid lumen. This enzyme kinetics analysis provides a more complete characterization of chloride and pH dependence of O2 evolution activity than has been previously presented.

  6. The role of pH on the resistance of resting- and active anammox bacteria to NO2- inhibition.

    PubMed

    Carvajal-Arroyo, José M; Puyol, Daniel; Li, Guangbin; Sierra-Álvarez, Reyes; Field, Jim A

    2014-10-01

    The anaerobic oxidation of ammonium (anammox) uses nitrite as terminal electron acceptor. The nitrite can cause inhibition to the bacteria that catalyze the anammox reaction. The literature shows a great divergence on the levels of NO2 (-) causing inhibition. Moreover, the conditions influencing the resistance of anammox bacteria to NO2 (-) inhibitory effect are not well understood. This work investigated the effect of the pH and the concentration of nitrite on the activity and metabolism of anammox granular sludge under different physiological conditions. Batch activity tests in a range of pH values were carried out in which either actively metabolizing cells or resting cells were exposed to nitrite in the presence or absence of the electron donating substrate ammonium, respectively. The response of the bacteria was evaluated by analyzing the specific anammox activity, the accumulation of nitric oxide, and the evolution of the ATP content in the biomass. Additionally, the effect of the pH on the tolerance of the biomass to single substrate feeding interruptions was evaluated in continuous anammox bioreactors. The results show that the influence of the pH on the NO2 (-) inhibition of anammox bacteria is greater under non-metabolizing conditions than during active metabolism. The exposure of resting cells to NO2 (-) (100 mg N L(-1) ) at pH values below 7.2 caused complete inhibition of the anammox activity. The inhibition was accompanied by accumulation of the intermediate, nitric oxide, in the gas phase. In contrast, just mild inhibition was observed for resting cells exposed to the same NO2 (-) concentration at pH values higher than 7.5 or any of the pH values tested in assays with actively metabolizing cells. ATP initially increased and subsequently decreased in time after resting cells were exposed to NO2 (-) suggesting an active response of the cells to nitrite stress. Furthermore, bioreactors operated at pH lower than 6.8 had greater sensitivity to NO2

  7. Changes in medullary extracellular pH, sympathetic and phrenic nerve activity during brainstem perfusion with CO2 enriched solutions.

    PubMed

    König, S A; Offner, B; Czachurski, J; Seller, H

    1995-01-20

    Measurements are presented of sympathetic nerve activity (SNA), phrenic nerve activity (PNA), and local extracellular pH (ECF pH) within the rostral ventrolateral medulla (RVLM) in response to perfusions of the RVLM with CO2-enriched saline. Experiments were performed on cats anaesthetized with chloralose. The ventrolateral medullary surface was exposed, and a catheter was placed in the left vertebral artery from the axilla to allow perfusion of the RVLM. Baroreceptor and peripheral chemoreceptor denervations were performed by cutting the vagal, aortic and carotid sinus nerves. The activities of the renal and the phrenic nerve were recorded, in some experiments in parallel with the cardiac nerve. Recordings of the pH were done with ion-sensitive theta-microelectrodes. A linear relationship between the CO2 concentration of the perfusate and the evoked changes in ECF pH was found. The ECF pH did not change systematically in one or the other direction within depths between 1 and 3 mm below the surface of the medulla. The various patterns of interaction of ECF pH, SNA, and PNA are described in detail. Phrenic nerve response to perfusions was very variable; a more prolonged increase in amplitude of phasic discharges compared to the duration of changes in SNA and ECF pH was the most frequent finding, but non-phasic tonic activation and complete silence were also seen during perfusions. SNA could also deviate from ECF pH both with regard to its latency and to its time course in response to perfusions. Therefore, this study provides further evidence for deviations of cardiorespiratory adaptation from ECF pH, corroborating the notion that this parameter is not the decisive one for central chemoreception.

  8. pH Measurement Using Dual-Wavelength Fluorescent Ratio by Two-Photon Excitation for Mitochondrial Activity

    NASA Astrophysics Data System (ADS)

    Kanazashi, Yasuaki; Li, Yongbo; Onojima, Takumi; Iwami, Kentaro; Ohta, Yoshihiro; Umeda, Norihiro

    2012-11-01

    A mitochondrion has a pH gradient between the two sides of its inner membrane in order to produce adenosine triphosphate (ATP). Because ATP depletion causes numerous diseases, the measurement of the pH value around the mitochondrion is expected to clarify the mechanism of these diseases. In this study, a dual-wavelength pH-sensitive dye was excited by two-photon absorption initiated using a femtosecond pulse laser. In addition, fluorescence from the dye was directly collected from the fluorescent point using the collection-mode probe of a scanning near-field optical microscope. By this proposed method, a pH calibration curve was obtained from the fluorescent intensity ratio of the dye solution, and temporal pH variations with 0.1 s time resolution following the addition of acid were observed. Moreover, mitochondrial activity on the basis of the pH changes was successfully observed in three different mitochondrial densities.

  9. Carbon, nitrogen and pH regulate the production and activity of a polygalacturonase isozyme produced by Penicillium expansum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The influence of carbon, nitrogen and pH on polygalacturonase activity produced by Penicillium expansum were investigated. P. expansum mycelial growth was greatest on lyophilized fruit tissue and the highest PG activity occurred in apple pectin medium. Nitrogen source influenced PG activity and was ...

  10. Continuous volatile fatty acid production from waste activated sludge hydrolyzed at pH 12.

    PubMed

    Yang, Xue; Wan, Chunli; Lee, Duu-Jong; Du, Maoan; Pan, Xiangliang; Wan, Fang

    2014-09-01

    This study adopted rapid alkaline treatment at pH 12 to hydrolyze 66% of total chemical oxygen demands. Then the hydrolyzed liquor was fermented in a continuous-flow stirred reactor to produce volatile fatty acids (VFAs) at 8-h hydraulic retention time and at 35 °C. The maximum VFA productivity reached 365 mg VFAs g(-1) volatile suspended solids in a 45-d operation, with most produced VFAs being acetate and propionate, principally produced by protein degradation. The Bacteroidia, ε-proteobacteria and the Clostridia were identified to be the classes correlating with the fermentation processes. The fermented liquor was applied to denitrifying phosphorus removal process as alternative carbon source after excess phosphorus and nitrogen being recycled via struvite precipitation. Fermented liquors from alkaline hydrolysis-acid fermentation on waste activated sludge are a potential renewable resource for applications that need organic carbons.

  11. The Mechanism by Which a Propeptide-encoded pH Sensor Regulates Spatiotemporal Activation of Furin*

    PubMed Central

    Williamson, Danielle M.; Elferich, Johannes; Ramakrishnan, Parvathy; Thomas, Gary; Shinde, Ujwal

    2013-01-01

    The proprotein convertase furin requires the pH gradient of the secretory pathway to regulate its multistep, compartment-specific autocatalytic activation. Although His-69 within the furin prodomain serves as the pH sensor that detects transport of the propeptide-enzyme complex to the trans-Golgi network, where it promotes cleavage and release of the inhibitory propeptide, a mechanistic understanding of how His-69 protonation mediates furin activation remains unclear. Here we employ biophysical, biochemical, and computational approaches to elucidate the mechanism underlying the pH-dependent activation of furin. Structural analyses and binding experiments comparing the wild-type furin propeptide with a nonprotonatable His-69 → Leu mutant that blocks furin activation in vivo revealed protonation of His-69 reduces both the thermodynamic stability of the propeptide as well as its affinity for furin at pH 6.0. Structural modeling combined with mathematical modeling and molecular dynamic simulations suggested that His-69 does not directly contribute to the propeptide-enzyme interface but, rather, triggers movement of a loop region in the propeptide that modulates access to the cleavage site and, thus, allows for the tight pH regulation of furin activation. Our work establishes a mechanism by which His-69 functions as a pH sensor that regulates compartment-specific furin activation and provides insights into how other convertases and proteases may regulate their precise spatiotemporal activation. PMID:23653353

  12. Effect of stress-induced high post-mortem pH on protease activity and tenderness of beef.

    PubMed

    Beltrán, J A; Jaime, I; Santolaria, P; Sañudo, C; Albertí, P; Roncalés, P

    1997-02-01

    Forty-four Swiss Brown young bulls were stressed by regrouping unfamiliar animals before slaughter. M. longissimus thoracis (6-9th ribs) of carcasses were analysed for post-mortem pH, protease activities (m- and α-calpain, calpastatin and cathepsin B + L), Warner-Bratzler shear force and sensory tenderness and juiciness. Muscles were classified into three groups, according to ultimate pH values: > 6.3, 6.3-5.8 and < 5.8. The most significant difference related to high pH was a higher activity of m-calpain at 7th day post mortem. It was also found that meat showing the highest pH was significantly more tender and juicy. Sensory tenderness was highly correlated with activity of m-calpain at 7th day post mortem (r = 0.776) and with ultimate pH (r = 0.708). It is concluded that high ultimate pH induced by stress significantly increases m-calpain activity, and this results in a greatly enhanced tenderisation of beef meat.

  13. The pH dependence of hairpin ribozyme catalysis reflects ionization of an active site adenine.

    PubMed

    Cottrell, Joseph W; Scott, Lincoln G; Fedor, Martha J

    2011-05-20

    Understanding how self-cleaving ribozymes mediate catalysis is crucial in light of compelling evidence that human and bacterial gene expression can be regulated through RNA self-cleavage. The hairpin ribozyme catalyzes reversible phosphodiester bond cleavage through a mechanism that does not require divalent metal cations. Previous structural and biochemical evidence implicated the amidine group of an active site adenosine, A38, in a pH-dependent step in catalysis. We developed a way to determine microscopic pK(a) values in active ribozymes based on the pH-dependent fluorescence of 8-azaadenosine (8azaA). We compared the microscopic pK(a) for ionization of 8azaA at position 38 with the apparent pK(a) for the self-cleavage reaction in a fully functional hairpin ribozyme with a unique 8azaA at position 38. Microscopic and apparent pK(a) values were virtually the same, evidence that A38 protonation accounts for the decrease in catalytic activity with decreasing pH. These results implicate the neutral unprotonated form of A38 in a transition state that involves formation of the 5'-oxygen-phosphorus bond.

  14. RX-26-AW/AX rifle bullet tests

    SciTech Connect

    Sharp, D.D.

    1980-04-01

    A series of rifle bullet tests was performed at the request of LLL for two explosives, RX-26-AW and RX-26-AX, using the Pantex version of the Picatinny Arsenal test (PA-2). Both exposives tested displayed essentially the same low sensitivity to bullet impact. Slow burning of the explosive was the highest order of reaction observed. Also, in a continuing effort to improve the capability of the test facility, different rifle sighting and shot alignment procedures were implemented during this test series.

  15. Guided Multiple Launch Rocket System/Guided Multiple Launch Rocket System Alternative Warhead (GMLRS/GMLRS AW)

    DTIC Science & Technology

    2013-12-01

    Unit Cost BA - Budget Authority/Budget Activity BY - Base Year DAMIR - Defense Acquisition Management Information Retrieval Dev Est - Development...Strategey for GMLRS AW, signed on January 7, 2013, states the program will conduct the Inital Operational Test and Evaluation ( IOT &E) during the Engineering...needed. Necessary assets will be procured to support IOT &E during EMD. The Current Total LRIP reported in the December 31, 2012 SAR was 4943. This

  16. Effect of pH on the Activity of Some Respiratory Inhibitors.

    PubMed

    Chin, C K; Lee, M

    1980-12-01

    Inhibition of respiration of cultured cells of Petunia hybrida by NaF, NaN(3), malonic acid, and salicylhydroxamic acid increased at low pH. This increase could be partially reversed by raising the pH of the medium. Uptake experiments show that the greater inhibition of malonic acid at low pH was not the result of greater uptake. The results suggest that the increase in inhibition at low pH could not be attributed to greater cell penetration.

  17. pH Effects on solubility, zeta potential, and correlation between antibacterial activity and molecular weight of chitosan.

    PubMed

    Chang, Shun-Hsien; Lin, Hong-Ting Victor; Wu, Guan-James; Tsai, Guo Jane

    2015-12-10

    Six chitosans with molecular weights (MWs) of 300, 156, 72.1, 29.2, 7.1, and 3.3 kDa were prepared by cellulase degradation of chitosan (300 kDa) and ultrafiltration techniques. We examined the correlation between activity against Escherichia coli and Staphylococcus aureus and chitosan MW, and provided the underlying explanation. In acidic pH conditions, the chitosan activity increased with increasing MW, irrespective of the temperature and bacteria tested. However, at neutral pH, chitosan activity increased as the MW decreased, and little activity was observed for chitosans with MW >29.2 kDa. At pH 5.0 and 6.0, chitosans exhibited good water solubility and zeta potential (ZP) decreased with the MW, whereas the solubility and ZP of the chitosans decreased with increasing MW at pH 7.0. Particularly, low solubility and negative ZP values were determined for chitosans with MW >29.2 kDa, which may explain the loss of their antibacterial activity at pH 7.0.

  18. Description of Aspergillus flavus growth under the influence of different factors (water activity, incubation temperature, protein and fat concentration, pH, and cinnamon essential oil concentration) by kinetic, probability of growth, and time-to-detection models.

    PubMed

    Kosegarten, Carlos E; Ramírez-Corona, Nelly; Mani-López, Emma; Palou, Enrique; López-Malo, Aurelio

    2017-01-02

    A Box-Behnken design was used to determine the effect of protein concentration (0, 5, or 10g of casein/100g), fat (0, 3, or 6g of corn oil/100g), aw (0.900, 0.945, or 0.990), pH (3.5, 5.0, or 6.5), concentration of cinnamon essential oil (CEO, 0, 200, or 400μL/kg) and incubation temperature (15, 25, or 35°C) on the growth of Aspergillus flavus during 50days of incubation. Mold response under the evaluated conditions was modeled by the modified Gompertz equation, logistic regression, and time-to-detection model. The obtained polynomial regression models allow the significant coefficients (p<0.05) for linear, quadratic and interaction effects for the Gompertz equation's parameters to be identified, which adequately described (R(2)>0.967) the studied mold responses. After 50days of incubation, every tested model system was classified according to the observed response as 1 (growth) or 0 (no growth), then a binary logistic regression was utilized to model A. flavus growth interface, allowing to predict the probability of mold growth under selected combinations of tested factors. The time-to-detection model was utilized to estimate the time at which A. flavus visible growth begins. Water activity, temperature, and CEO concentration were the most important factors affecting fungal growth. It was observed that there is a range of possible combinations that may induce growth, such that incubation conditions and the amount of essential oil necessary for fungal growth inhibition strongly depend on protein and fat concentrations as well as on the pH of studied model systems. The probabilistic model and the time-to-detection models constitute another option to determine appropriate storage/processing conditions and accurately predict the probability and/or the time at which A. flavus growth occurs.

  19. Sorption of triclosan onto activated carbon, kaolinite and montmorillonite: effects of pH, ionic strength, and humic acid.

    PubMed

    Behera, Shishir Kumar; Oh, Seok-Young; Park, Hung-Suck

    2010-07-15

    Sorption of triclosan on three sorbents, viz., activated carbon, kaolinite and montmorillonite was studied as a function of pH, ionic strength and humic acid (HA) concentration through controlled batch experiments. Triclosan sorption was found to be higher in the acidic pH range, as varying pH showed significant influence on the surface charge of the sorbents and degree of ionization of the sorbate. Sorption capacity of the sorbents increased with an increase in the ionic strength of solution. At low pH (pH 3), the overall increase in triclosan sorption was 1.2, approximately 4 and 3.5 times, respectively for activated carbon, kaolinite and montmorillonite when ionic strength was increased from 1x10(-3) to 5x10(-1) M. Triclosan sorption onto activated carbon decreased from 31.4 to 10.6 mg g(-1) by increasing the HA concentration to 200 mg C L(-1). However, during sorption onto kaolinite and montmorillonite, the effect of HA was very complex probably due to (i) hydrophobicity (log K(ow)=4.76) of triclosan; and (ii) complexation of HA with triclosan. Though triclosan sorption onto activated carbon is higher, the potential of kaolinite and montmorillonite in controlling the transport of triclosan in subsurface environment can still be appreciable.

  20. Active auxin uptake by zucchini membrane vesicles: quantitation using ESR volume and delta pH determinations

    SciTech Connect

    Lomax, T.L.; Mehlhorn, R.J.; Briggs, W.R.

    1985-10-01

    Closed and pH-tight membrane vesicles prepared from hypocotyls of 5-day-old dark-grown seedlings of Cucurbita pepo accumulate the plant growth hormone indole-3-acetic acid along an imposed proton gradient (pH low outside, high inside). The use of electron paramagnetic spin probes permitted quantitation both of apparent vesicle volume and magnitude of the pH gradient. Under the experimental conditions used, hormone accumulation was at minimum 20-fold, a value 4 times larger than what one would predict if accumulation reflected only diffusional equilibrium at the measured pH gradient. It is concluded that hormone uptake is an active process, with each protonated molecule of hormone accompanied by an additional proton. Experiments with ionophores confirm that it is the pH gradient itself which drives the uptake.

  1. Diiron species containing a cyclic P(Ph)2N(Ph)2 diphosphine related to the [FeFe]H2ases active site.

    PubMed

    Lounissi, Sondès; Capon, Jean-François; Gloaguen, Frédéric; Matoussi, Fatma; Pétillon, François Y; Schollhammer, Philippe; Talarmin, Jean

    2011-01-21

    A new dissymmetrically disubstituted diiron dithiolate species, [Fe(2)(CO)(4)(κ(2)-P(Ph)(2)N(Ph)(2))(μ-pdt)] (pdt = S(CH(2))(3)S), was prepared by using a flexible cyclic base-containing diphosphine, 1,3,5,7-tetraphenyl 1,5-diaza-3,7-diphosphacyclooctane (P(Ph)(2)N(Ph)(2) = {PhPCH(2)NPh}(2)). Preliminary investigations of proton and electron transfers on the diiron system have been done.

  2. SMYD3-mediated lysine methylation in the PH domain is critical for activation of AKT1

    PubMed Central

    Yoshioka, Yuichiro; Suzuki, Takehiro; Matsuo, Yo; Nakakido, Makoto; Tsurita, Giichiro; Simone, Cristiano; Watanabe, Toshiaki; Dohmae, Naoshi; Nakamura, Yusuke; Hamamoto, Ryuji

    2016-01-01

    AKT1 is a cytosolic serine/threonine kinase that is overexpressed in various types of cancer and has a central role in human tumorigenesis. Although it is known that AKT1 is post-translationally modified in various ways including phosphorylation and ubiquitination, methylation has not been reported so far. Here we demonstrate that the protein lysine methyltransferase SMYD3 methylates lysine 14 in the PH domain of AKT1 both in vitro and in vivo. Lysine 14-substituted AKT1 shows significantly lower levels of phosphorylation at threonine 308 than wild-type AKT1, and knockdown of SMYD3 as well as treatment with a SMYD3 inhibitor significantly attenuates this phosphorylation in cancer cells. Furthermore, substitution of lysine 14 diminishes the plasma membrane accumulation of AKT1, and cancer cells overexpressing lysine 14-substiuted AKT1 shows lower growth rate than those overexpressing wild-type AKT1. These results imply that SMYD3-mediated methylation of AKT1 at lysine 14 is essential for AKT1 activation and that SMYD3-mediated AKT1 methylation appears to be a good target for development of anti-cancer therapy. PMID:27626683

  3. 241-AW Tank Farm Construction Extent of Condition Review for Tank Integrity

    SciTech Connect

    Barnes, Travis J.; Gunter, Jason R.; Reeploeg, Gretchen E.

    2013-11-19

    This report provides the results of an extent of condition construction history review for the 241-AW tank farm. The construction history of the 241-AW tank farm has been reviewed to identify issues similar to those experienced during tank AY-102 construction. Those issues and others impacting integrity are discussed based on information found in available construction records, using tank AY-102 as the comparison benchmark. In the 241-AW tank farm, the fourth double-shell tank farm constructed, similar issues as those with tank 241-AY-102 construction occured. The overall extent of similary and affect on 241-AW tank farm integrity is described herein.

  4. Effect of pH and Temperature on Enzyme Activity of Chitosanase Produced Under Solid Stated Fermentation by Trichoderma spp.

    PubMed

    da Silva, Luis C A; Honorato, Talita L; Cavalcante, Rosane S; Franco, Telma T; Rodrigues, Sueli

    2012-03-01

    Trichoderma strains were extensively studied as biocontrol agents due to their ability of producing hydrolytic enzymes, which are considered key enzymes because they attack the insect exoskeleton allowing the fungi infection. The present work aimed to evaluate the ability of chitosanase production by four Trichoderma strains (T. harzianum, T. koningii, T. viride and T. polysporum) under solid stated fermentation and to evaluate the effect of pH and temperature on enzyme activity. pH strongly affected the enzyme activity from all tested strains. Chitosanase from T. harzianum and T. viride presented optimum activity at pH 5.0 and chitosanase from T. koningii and T. polysporum presented optimum activity at pH 5.5. Temperature in the range of 40-50°C did not affect enzyme activity. T. polysporum was found as the most promising strain to produce chitosanase with maximal enzyme activity of about 1.4 IU/gds, followed by T. viride (~1.2 IU/gds) and T. harzianum (1.06 IU/gds).

  5. Detecting activity-evoked pH changes in human brain.

    PubMed

    Magnotta, Vincent A; Heo, Hye-Young; Dlouhy, Brian J; Dahdaleh, Nader S; Follmer, Robin L; Thedens, Daniel R; Welsh, Michael J; Wemmie, John A

    2012-05-22

    Localized pH changes have been suggested to occur in the brain during normal function. However, the existence of such pH changes has also been questioned. Lack of methods for noninvasively measuring pH with high spatial and temporal resolution has limited insight into this issue. Here we report that a magnetic resonance imaging (MRI) strategy, T(1) relaxation in the rotating frame (T(1)ρ), is sufficiently sensitive to detect widespread pH changes in the mouse and human brain evoked by systemically manipulating carbon dioxide or bicarbonate. Moreover, T(1)ρ detected a localized acidosis in the human visual cortex induced by a flashing checkerboard. Lactate measurements and pH-sensitive (31)P spectroscopy at the same site also identified a localized acidosis. Consistent with the established role for pH in blood flow recruitment, T(1)ρ correlated with blood oxygenation level-dependent contrast commonly used in functional MRI. However, T(1)ρ was not directly sensitive to blood oxygen content. These observations indicate that localized pH fluctuations occur in the human brain during normal function. Furthermore, they suggest a unique functional imaging strategy based on pH that is independent of traditional functional MRI contrast mechanisms.

  6. Compression dewatering of municipal activated sludge: effects of salt and pH.

    PubMed

    Raynaud, Mickael; Vaxelaire, Jean; Olivier, Jérémy; Dieudé-Fauvel, Emilie; Baudez, Jean-Christophe

    2012-09-15

    Even after mechanical dewatering, activated sludge contains a large amount of water. Due to its composition and biological nature this material is usually highly compressible and known to be difficult to dewater. In the present work, two treatments (salt addition and pH modification) are proposed to highlight some aspects which could explain the poor dewaterability of activated sludge. Dewatering tests are carried out in a pressure-driven device in order to well examine both, filtration and compression stages. Physico-chemical parameters, such as surface charge, hydrophobicity, extracellular polymeric substances (EPS) content and filtrate turbidity are measured on the tested sludge, for a better analysis of dewatering results. The dewatering ability of the sludge is widely linked to the cohesion of the flocculated matrix and the presence of fine particles. Both treatments alter the flocculated matrix and release fine particles. The release of fine particles tends to clog both, the filter cake and the filter medium. Consequently, the filtration rate decreases due to higher resistances to the flow. On another hand, the polymeric matrix breakdown enables to release some water trapped within the floc to the bulk liquid phase and thus facilitates its removal, which tends to decrease the moisture content of the filter-cake. It also impacts the compression dewatering step. The more destroyed structures lead to less elastic cakes and thus a slower primary consolidation stage. At the opposite, the mobility of the broken aggregates within the filter-cake does not seem to be improved by size reduction (the kinetics of the secondary consolidation stage are not significantly modified).

  7. Targetable kinase-activating lesions in Ph-like acute lymphoblastic leukemia | Office of Cancer Genomics

    Cancer.gov

    Publication Abstract:  Philadelphia chromosome-like acute lymphoblastic leukemia (Ph-like ALL) is characterized by a gene-expression profile similar to that of BCR-ABL1-positive ALL, alterations of lymphoid transcription factor genes, and a poor outcome. The frequency and spectrum of genetic alterations in Ph-like ALL and its responsiveness to tyrosine kinase inhibition are undefined, especially in adolescents and adults. We performed genomic profiling of 1725 patients with precursor B-cell ALL and detailed genomic analysis of 154 patients with Ph-like ALL.

  8. Influence of pH on the sonolysis of ciprofloxacin: Biodegradability, ecotoxicity and antibiotic activity of its degradation products.

    PubMed

    De Bel, Evelien; Dewulf, Jo; Witte, Bavo De; Van Langenhove, Herman; Janssen, Colin

    2009-09-01

    The presence of antibiotics in the aquatic environment has raised concerns due to the potential risk for the emergence or persistence of antibiotic resistance. Antibiotics are often poorly degraded in conventional wastewater treatment plants. In this study, sonolysis at 520 kHz and 92 W L(-1) was used for the degradation of the fluoroquinolone antibiotic ciprofloxacin. In a first experiment at pH 7, 57% of the ciprofloxacin (15 mg L(-1)) was degraded after 120 min of ultrasonic irradiation at 25 degrees C. pH proved to be an important parameter determining the degradation rate, since the pseudo first order degradation constant increased almost fourfold when comparing treatment at pH 7 (0.0058 min(-1)) and pH 10 (0.0069 min(-1)) with that at pH 3 (0.021 min(-1)). This effect can be attributed to the degree of protonation of the ciprofloxacin molecule. The BOD/COD ratio of the solutions, which is a measure for their biodegradability, increased from 0.06 to 0.60, 0.17, and 0.18 after 120 min of irradiation depending on the pH (3, 7, and 10, respectively). The solution treated at pH 3 can even be considered readily biodegradable (BOD/COD>0.4). The antibiotic activity against Escherichia coli (G-) and Bacillus coagulans (G+) of the treated solutions also reduced after sonolysis. The highest decrease was again found when irradiated at pH 3. In contrast, ecotoxicity of the solutions to the alga Pseudokirchneriella subcapitata increased 3- to 10-fold after 20 min of treatment, suggesting the formation of toxic degradation products. The toxicity slowly diminished during further treatment.

  9. Approach on quantitative structure-activity relationship for design of a pH neutral carrier containing tertiary amino group.

    PubMed

    Cao, Zhong; Gong, Fu-Chun; Li, He-Ping; Xiao, Zhong-Liang; Long, Shu; Zhang, Ling; Peng, San-Jun

    2007-01-02

    The quantitative structure-activity relationship (QSAR) for neutral carriers used to prepare hydrogen ion sensors has been studied. A series of synthesized carrier compounds were taken as the training set. Five molecular structure parameters of the compounds were calculated by using CNDO/2 algorithm and used as feature variables in constructing QSAR model. The lower and upper limits of the linear pH response range were taken as the activity measure. The corresponding model equations were derived from the stepwise regression procedure. With the established QSAR model, a new pH carrier, (4-hydroxybenzyl) didodecylamine (XIII) was proposed and synthesized. The PVC membrane pH electrode based on carrier XIII with a wide pH linear response range of 2.0-12.5 was prepared. Having a theoretical Nernstian response slope of 57.2+/-0.3 mV/pH (n=5 at 25 degrees C) without a super-Nernstian phenomenon, the sensor had low resistance, short response time, high selectivity and good reproducibility. Moreover, the sensor was successfully applied to detecting the pH value of serum samples.

  10. Astronomy popularization at the University of Wrocław

    NASA Astrophysics Data System (ADS)

    Preś, Paweł; Cader-Sroka, Barbara; Berlicki, Arkadiusz

    2016-06-01

    Science popularization found many expressions in the history of the Astronomical Institute of the University of Wrocław. The long-lasting is the tradition of popular astronomy lectures. Occasional celestial events are the base of public observations. Since 1998 the Institute boldly participates in the Lower-Silesian Science Festival. The rising public interest in astronomy encouraged us to establish ''Planetarium'' laboratory in 2008. Together with the Astronomical Institute of the Academy of Sciences of the Czech Republic we established in 2009 the very first trans-border dark-sky park in the location of Izera Mountains, where since then the public can benefit from excellent observational conditions and the astronomers' support.

  11. Multitaxon activity profiling reveals differential microbial response to reduced seawater pH and oil pollution.

    PubMed

    Coelho, Francisco J R C; Cleary, Daniel F R; Costa, Rodrigo; Ferreira, Marina; Polónia, Ana R M; Silva, Artur M S; Simões, Mário M Q; Oliveira, Vanessa; Gomes, Newton C M

    2016-09-01

    There is growing concern that predicted changes to global ocean chemistry will interact with anthropogenic pollution to significantly alter marine microbial composition and function. However, knowledge of the compounding effects of climate change stressors and anthropogenic pollution is limited. Here, we used 16S and 18S rRNA (cDNA)-based activity profiling to investigate the differential responses of selected microbial taxa to ocean acidification and oil hydrocarbon contamination under controlled laboratory conditions. Our results revealed that a lower relative abundance of sulphate-reducing bacteria (Desulfosarcina/Desulfococcus clade) due to an adverse effect of seawater acidification and oil hydrocarbon contamination (reduced pH-oil treatment) may be coupled to changes in sediment archaeal communities. In particular, we observed a pronounced compositional shift and marked reduction in the prevalence of otherwise abundant operational taxonomic units (OTUs) belonging to the archaeal Marine Benthic Group B and Marine Hydrothermal Vent Group (MHVG) in the reduced pH-oil treatment. Conversely, the abundance of several putative hydrocarbonoclastic fungal OTUs was higher in the reduced pH-oil treatment. Sediment hydrocarbon profiling, furthermore, revealed higher concentrations of several alkanes in the reduced pH-oil treatment, corroborating the functional implications of the structural changes to microbial community composition. Collectively, our results advance the understanding of the response of a complex microbial community to the interaction between reduced pH and anthropogenic pollution. In future acidified marine environments, oil hydrocarbon contamination may alter the typical mixotrophic and k-/r-strategist composition of surface sediment microbiomes towards a more heterotrophic state with lower doubling rates, thereby impairing the ability of the ecosystem to recover from acute oil contamination events.

  12. The enzymatic activities of the Escherichia coli basic aliphatic amino acid decarboxylases exhibit a pH zone of inhibition.

    PubMed

    Kanjee, Usheer; Gutsche, Irina; Ramachandran, Shaliny; Houry, Walid A

    2011-11-01

    The stringent response regulator ppGpp has recently been shown by our group to inhibit the Escherichia coli inducible lysine decarboxylase, LdcI. As a follow-up to this observation, we examined the mechanisms that regulate the activities of the other four E. coli enzymes paralogous to LdcI: the constitutive lysine decarboxylase LdcC, the inducible arginine decarboxylase AdiA, the inducible ornithine decarboxylase SpeF, and the constitutive ornithine decarboxylase SpeC. LdcC and SpeC are involved in cellular polyamine biosynthesis, while LdcI, AdiA, and SpeF are involved in the acid stress response. Multiple mechanisms of regulation were found for these enzymes. In addition to LdcI, LdcC and SpeC were found to be inhibited by ppGpp; AdiA activity was found to be regulated by changes in oligomerization, while SpeF and SpeC activities were regulated by GTP. These findings indicate the presence of multiple mechanisms regulating the activity of this important family of decarboxylases. When the enzyme inhibition profiles are analyzed in parallel, a "zone of inhibition" between pH 6 and pH 8 is observed. Hence, the data suggest that E. coli utilizes multiple mechanisms to ensure that these decarboxylases remain inactive around neutral pH possibly to reduce the consumption of amino acids at this pH.

  13. NHE1 is the sodium-hydrogen exchanger active in acute intracellular pH regulation in preimplantation mouse embryos.

    PubMed

    Siyanov, Violetta; Baltz, Jay M

    2013-06-01

    Sodium-hydrogen exchangers (NHE) of the Slc9 gene family are the major regulators of intracellular pH against acidosis in mammalian cells. Of five plasma membrane NHE isoforms, mouse oocytes and preimplantation embryos express mRNAs encoding NHE1 (SLC9A1), NHE3 (SLC9A3), and NHE4 (SLC9A4), with higher mRNA levels for each in oocytes through one-cell stage embryos and lower levels after the two-cell stage. NHE2 (SLC9A2) and NHE5 (SLC9A5) are not expressed. Measurements of intracellular pH during recovery from induced acidosis indicated that recovery occurred via NHE activity at all preimplantation stages assessed (one-cell, two-cell, eight-cell and morula). Recovery from acidosis at each stage was entirely inhibited by cariporide, which is very highly selective for NHE1. In contrast, the moderately NHE3-selective inhibitor S3226 did not preferentially block recovery, nor did adding S3226 increase inhibition over cariporide alone, indicating that NHE3 did not play a role. There was no indication of NHE4 activity. Another regulator of intracellular pH against acidosis, the sodium-dependent bicarbonate/chloride exchanger (NDBCE; SLC4A8), had low or absent activity in two-cell embryos. Thus, NHE1 appears to be the only significant regulator of intracellular pH in preimplantation mouse embryos. Culturing embryos from the one-cell or two-cell stages in acidotic medium inhibited their development. Unexpectedly, inhibition of NHE1 with cariporide, NDBCE with DIDS, or both together did not affect embryo development to the blastocyst stage more substantially under conditions of chronic acidosis than at normal pH. Preimplantation mouse embryos thus appear to have limited capacity to resist chronic acidosis using intracellular pH regulatory mechanisms.

  14. Influence of copper, manganese and pH on the growth and several enzyme activities in mycorrhizal fungus Amanita muscaria.

    PubMed

    Kong, E X

    1995-01-01

    The effects of various concentrations of copper, manganese and pH on the growth and several enzyme activities of mycorrhizal fungus Amanita muscaria were investigated. Cu (5-25 mg l-1) and lower pH (3.0-4.0) strongly inhibited the mycelial growth (dry weight), however, the protein content was not affected evidently. Some enzyme activities were lower as the Cu and Mn concentrations were higher and other enzymes had the maximum values at the specified concentration. The activities of the following enzymes were significantly correlated with the fungal growth after the treatment with Cu: G6PDH, MTLDH and trehalase, and with Mn: G6PDH, MTLDH and alpha-mannosidase respectively. Measurement of these enzyme activities might provide a useful biochemical criterion for the evaluation of the fungitoxicity of soil contaminated by copper or manganese.

  15. 47 CFR 27.1160 - Cost-sharing requirements for AWS.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... benefit from the clearance of this spectrum by other AWS entities or by a voluntarily relocating microwave... specified in §§ 22.602 and 101.69 of this chapter. If an AWS licensee enters into a spectrum leasing arrangement (as set forth in part 1, subpart X of this chapter) and the spectrum lessee triggers a...

  16. 47 CFR 27.1160 - Cost-sharing requirements for AWS.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... benefit from the clearance of this spectrum by other AWS entities or by a voluntarily relocating microwave... specified in §§ 22.602 and 101.69 of this chapter. If an AWS licensee enters into a spectrum leasing arrangement (as set forth in part 1, subpart X of this chapter) and the spectrum lessee triggers a...

  17. 47 CFR 27.1160 - Cost-sharing requirements for AWS.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... benefit from the clearance of this spectrum by other AWS entities or by a voluntarily relocating microwave... specified in § 22.602 and 101.69 of this chapter. If an AWS licensee enters into a spectrum leasing arrangement (as set forth in part 1, subpart X of this chapter) and the spectrum lessee triggers a...

  18. The effects of pH and various salts upon the activity of a series of superoxide dismutases.

    PubMed

    O'Neill, P; Davies, S; Fielden, E M; Calabrese, L; Capo, C; Marmocchi, F; Natoli, G; Rotilio, G

    1988-04-01

    The CuZn superoxide dismutases (SODs) from ox, sheep, pig and yeast were investigated by pulse radiolysis in order to evaluate the role of electrostatic interactions between O2.- and SOD proteins in the mechanism of action of the SOD enzymes. The protein net charge in this series varies, as evaluated by the protein pI values spanning over a large range of pH: 8.0 (sheep), 6.5 (pig), 5.2 (ox) and 4.6 (yeast). The amino acid sequences are largely conserved, with the three mammalian proteins being highly homologous and the yeast protein having some distinct variations in the region surrounding the active site. At pH 8.0 the activities of the SODs from various sources are similar, though the minor differences observed suggest that in the highly homologous mammalian series the most acidic protein is the most enzymically efficient one. The pH-dependences of the various activities in the pH range 7-12 are similar, and the related curves are best fitted by two pK values, which are approx. 9.2 and 11.0 for the mammalian enzymes and 9.1 and 11.4 for the yeast enzyme. The activities of the proteins at I 0.1 are decreased by approx. 20% when compared with the activity at I 0.02 at pH 8.5, whereas at pH above 10 the pH-dependence of the activity approaches that determined at I 0.02 and at pH 11.9 the activity is essentially independent of ionic strength. The dependence upon ionic strength also depends on the salt used, with perchlorate being more effective than phosphate or borate or Mops and still effective at pH above 10.5, where the effect of other salts becomes negligible. The dual and concerted dependence of the activities of different SODs on pH and salt concentration is explained with the encounter of O2.- with the active-site copper being governed by the protonation of two positively charged groups in the vicinity of the active site. The gradient between these localized charges and the rest of the protein may explain the different activities of the mammalian proteins at

  19. Improved methane production from waste activated sludge with low organic content by alkaline pretreatment at pH 10.

    PubMed

    Feng, L Y; Yang, L Q; Zhang, L X; Chen, H L; Chen, J

    2013-01-01

    Sludge with low organic content always results in an unsatisfactory performance, even failure of anaerobic digestion. The alkaline pretreatment effect on anaerobic digestion of sludge with low organic content has seldom been studied although it gives many benefits for sludge with high organic content. In this study the influence of alkaline pretreatment (pH 10, an effective alkaline pH) on the solubilization and methane production from waste activated sludge (WAS) with low organic content was investigated. Results from biochemical methane potential (BMP) experiments showed that anaerobic biodegradability of WAS was greatly improved by alkaline pretreatment at pH 10. Methane production from the current WAS under conditions of pretreatment time 4 h and digestion time 15 d was 139.6 mL/g VS (volatile solids), much higher than that from the unpretreated WAS with digestion time of 20 d (75.2 mL/g VS). Also, the solubilization of WAS was significantly accelerated by alkaline pretreatment. Mechanism exploration indicated that the general activities of anaerobic microorganisms, specific activities of key enzymes and the amounts of methanogens were enhanced by alkaline pretreatment at pH 10, showing good agreement with methane production.

  20. Cloning, Overexpression, and Characterization of a Metagenome-Derived Phytase with Optimal Activity at Low pH.

    PubMed

    Tan, Hao; Wu, Xiang; Xie, Liyuan; Huang, Zhongqian; Gan, Bingcheng; Peng, Weihong

    2015-06-01

    A phytase gene was identified in a publicly available metagenome derived from subsurface groundwater, which was deduced to encode for a protein of the histidine acid phosphatase (HAP) family. The nucleotide sequence of the phytase gene was chemically synthesized and cloned, in order to further overexpress the phytase in Escherichia coli. Purified protein of the recombinant phytase demonstrated an activity for phytic acid of 298 ± 17 μmol P/min/mg, at the pH optimum of 2.0 with the temperature of 37 °C. Interestingly, the pH optimum of this phytase is much lower in comparison with most HAP phytases known to date. It suggests that the phytase could possess improved adaptability to the low pH condition caused by the gastric acid in livestock and poultry stomachs.

  1. Effects of pH and inorganic salts on the adsorption of phenol from aqueous systems on activated decolorizing charcoal

    SciTech Connect

    Halhouli, K.A.; Darwish, N.A.; Al-Dhoon, N.M.

    1995-10-01

    An experimental investigation of the effects of pH and three inorganic salts (KCl, KI, and NaCl) on the adsorption isotherms of phenol (from a dilute aqueous solution) on activated charcoal was conducted. Each salt was studied at three different concentrations, i.e., 0.1, 0.01, and 0.005 M. The effect of pH (in the pH range 3 to 11) in the presence of KI, KCl, and NaCl was also investigated. The concentration of phenol in the aqueous systems studied ranged from 10 to 200 ppm. The temperature effect was also studied, and the resulting experimental equilibrium isotherms at 30, 40, and 55{degrees}C are well represented by Freundlich, Langmuir, and Redlich-Paterson isotherms. The relevant parameters for these isotherms are presented.

  2. Alternative exon-encoding regions of Locusta migratoria muscle myosin modulate the pH dependence of ATPase activity.

    PubMed

    Li, J; Lu, Z; He, J; Chen, Q; Wang, X; Kang, L; Li, X-D

    2016-12-01

    Whereas the vertebrate muscle myosin heavy chains (MHCs) are encoded by a family of Mhc genes, most insects examined to date contain a single Mhc gene and produce all of the different MHC isoforms by alternative RNA splicing. Here, we found that the migratory locust, Locusta migratoria, has one Mhc gene, which contains 41 exons, including five alternative exclusive exons and one differently included penultimate exon, and potentially encodes 360 MHC isoforms. From the adult L. migratoria, we identified 14 MHC isoforms (including two identical isoforms): four from flight muscle (the thorax dorsal longitudinal muscle), three from jump muscle (the hind leg extensor tibiae muscle) and seven from the abdominal intersegmental muscle. We purified myosins from flight muscle and jump muscle and characterized their motor activities. At neutral pH, the flight and the jump muscle myosins displayed similar levels of in vitro actin-gliding activity, whereas the former had a slightly higher actin-activated ATPase activity than the latter. Interestingly, the pH dependences of the actin-activated ATPase activity of these two myosins are different. Because the dominant MHC isoforms in these two muscles are identical except for the two alternative exon-encoding regions, we propose that these two alternative regions modulate the pH dependence of L. migratoria muscle myosin.

  3. The effect of pH on polymorph formation of the pharmaceutically active compound tianeptine.

    PubMed

    Orola, Liana; Veidis, Mikelis V; Sarcevica, Inese; Actins, Andris; Belyakov, Sergey; Platonenko, Aleksandrs

    2012-08-01

    The anti-depressant pharmaceutical tianeptine has been investigated to determine the dynamics of polymorph formation under various pH conditions. By varying the pH two crystalline polymorphs were isolated. The molecular and crystal structures have been determined to identify the two polymorphs. One polymorph is an amino carboxylic acid and the other polymorph is a zwitterion. In the solid state the tianeptine moieties are bonded through hydrogen bonds. The zwitterion was found to be less stable and transformed to the acid form. During this investigation an amorphous form was identified.

  4. AW00179 potentiates TRAIL-mediated death of human lung cancer H1299 cells through ROS-JNK-c-Jun-mediated up-regulation of DR5 and down-regulation of anti-apoptotic molecules.

    PubMed

    Hwang, Mi-Kyung; Ryu, Byung Jun; Kim, Seong Hwan

    2012-10-01

    Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) triggers apoptosis in tumor cells, but when used alone, it is not effective at treating TRAIL-resistant tumors. This resistance is challenging for TRAIL-based anti-cancer therapies. In this study, we found that 1-(4-trifluoromethoxy-phenyl)-3-[4-(5-trifluoromethyl-2,5-dihydro-pyrazol-1-yl)-phenyl]-urea (AW00179) sensitized human lung cancer H1299 cells to TRAIL-mediated apoptosis. Even in the absence of TRAIL, AW00179 strongly induced DR5 expression and decreased the expression of anti-apoptotic proteins, suggesting that the sensitizing effect of AW00179 on TRAIL-mediated apoptosis is due to increased levels of DR5 protein and decreased anti-apoptotic molecules. AW00179 also induced the activation of c-Jun and ERK; however, a pharmacologic inhibition study revealed that JNK-c-Jun signaling is involved in the induction of DR5 expression. In addition, reactive oxygen species (ROS) appear to be involved in AW00179 activity. In conclusion, AW00179 has the potential to sensitize H1299 cells to TRAIL-mediated apoptosis through two distinct mechanisms: ROS-JNK-c-Jun-mediated up-regulation of DR5, and down-regulation of anti-apoptotic molecules.

  5. Influence of Urinary pH on the Pharmacokinetics of Cinoxacin in Humans and on Antibacterial Activity In Vitro

    PubMed Central

    Barbhaiya, Rashmi H.; Gerber, Andreas U.; Craig, William A.; Welling, Peter G.

    1982-01-01

    The impact of acidification and alkalinization of the urine on the pharmacokinetics of cinoxacin was examined after single 500-mg oral doses were administered to nine healthy male volunteers. Acidic and alkaline conditions were achieved by repeated oral doses of ammonium chloride or sodium bicarbonate, respectively. Plasma cinoxacin levels in all subjects were adequately described in terms of one-compartment-model kinetics with first-order absorption and elimination. Acidification and alkalinization treatment had no effect on cinoxacin absorption or distribution. The mean elimination half-life of cinoxacin in plasma was 1.1, 2.0, and 0.6 h in control subjects and with acidification and alkalinization of urine, respectively. Recovery of intact cinoxacin in samples of urine collected 0 to 36 h after cinoxacin administration represented 65% of the dose in control subjects and urine acidification and 80% of the dose with alkalinization of urine. The mean renal clearance of cinoxacin was 76, 118, and 278 ml/min with acidification, control, and alkalinization, respectively, and renal clearance was highly correlated with urinary pH. Urine concentrations of cinoxacin were significantly higher with alkalinization compared with control values during the first 4 h after drug administration. Urine cinoxacin concentrations were reduced somewhat by acidification, but these tended not to be significantly different from control values. Changes in cinoxacin elimination owing to urine pH are less pronounced in humans than in dogs. The antibacterial activity of cinoxacin against some common urinary tract pathogens was pH dependent. A four- to eightfold reduction in cinoxacin activity was generally observed at pH 8 compared with lower pH values. However, in view of the high levels of cinoxacin which are obtained in both acidic and basic urine, the impact of urine pH on cinoxacin antibacterial efficacy would be of minor clinical importance. PMID:7103450

  6. Inter and intrasite analyses of cultural materials from U20aw, Nye County, Nevada

    SciTech Connect

    Hicks, P.A.; Pippin, L.C.; Henton, G.H.; Hemphill, M.L.; Lockett, C.L.

    1991-12-01

    In the Spring of 1986 Desert Research Institute (DRI) conducted a Class III archaeological survey of Drill Hole U20aw on the Nevada Test Site, Nye County, Nevada. Seven archaeological sites were located during the course of this survey including two temporary camps, four lithic scatters, and a possible pinyon cache. This report presents the results of the analyses of the data derived from all sites investigated during the data recovery operations on Drill Hole U20aw. Detailed analyses were focused on the spatial distribution of artifacts and features within and between sites in the southern portion of the study area (26Ny4867, 26Ny4869, and 26Ny4871). These analyses indicate that 26Ny4871 served principally as a temporary camp, while the area around the canyonhead to the east (which includes 26Ny4867 and 26Ny4869) seems to have been used as a site for both temporary camps and special activity loci. Projectile point styles suggest that the area was occupied from the Early Archaic into the early Historic period. Analyses of the artifacts that were recovered indicate that obsidian was the preferred material for all classes of flaked stone tools. All stages of lithic reduction are represented on the sites, but core reduction and thinning of bifaces appear to have been the primary activities. Processing of floral foods is indicated by the presence of several ground stone artifacts. Pinyon nuts and other items appear to have been stored in the area of 26Ny4869 and to the north of the drill hole as evidenced by the presence of several rock features that may have served as caches.

  7. Correlating hydrogen oxidation and evolution activity on platinum at different pH with measured hydrogen binding energy

    SciTech Connect

    Sheng, WC; Zhuang, ZB; Gao, MR; Zheng, J; Chen, JGG; Yan, YS

    2015-01-08

    The hydrogen oxidation/evolution reactions are two of the most fundamental reactions in distributed renewable electrochemical energy conversion and storage systems. The identification of the reaction descriptor is therefore of critical importance for the rational catalyst design and development. Here we report the correlation between hydrogen oxidation/evolution activity and experimentally measured hydrogen binding energy for polycrystalline platinum examined in several buffer solutions in a wide range of electrolyte pH from 0 to 13. The hydrogen oxidation/evolution activity obtained using the rotating disk electrode method is found to decrease with the pH, while the hydrogen binding energy, obtained from cyclic voltammograms, linearly increases with the pH. Correlating the hydrogen oxidation/evolution activity to the hydrogen binding energy renders a monotonic decreasing hydrogen oxidation/evolution activity with the hydrogen binding energy, strongly supporting the hypothesis that hydrogen binding energy is the sole reaction descriptor for the hydrogen oxidation/evolution activity on monometallic platinum.

  8. The effect of pH and natural microbial phosphatase activity on the speciation of uranium in subsurface soils

    NASA Astrophysics Data System (ADS)

    Beazley, Melanie J.; Martinez, Robert J.; Webb, Samuel M.; Sobecky, Patricia A.; Taillefert, Martial

    2011-10-01

    The biomineralization of U(VI) phosphate as a result of microbial phosphatase activity is a promising new bioremediation approach to immobilize uranium in both aerobic and anaerobic conditions. In contrast to reduced uranium minerals such as uraninite, uranium phosphate precipitates are not susceptible to changes in oxidation conditions and may represent a long-term sink for uranium in contaminated environments. So far, the biomineralization of U(VI) phosphate has been demonstrated with pure cultures only. In this study, two uranium contaminated soils from the Department of Energy Oak Ridge Field Research Center (ORFRC) were amended with glycerol phosphate as model organophosphate source in small flow-through columns under aerobic conditions to determine whether natural phosphatase activity of indigenous soil bacteria was able to promote the precipitation of uranium(VI) at pH 5.5 and 7.0. High concentrations of phosphate (1-3 mM) were detected in the effluent of these columns at both pH compared to control columns amended with U(VI) only, suggesting that phosphatase-liberating microorganisms were readily stimulated by the organophosphate substrate. Net phosphate production rates were higher in the low pH soil (0.73 ± 0.17 mM d -1) compared to the circumneutral pH soil (0.43 ± 0.31 mM d -1), suggesting that non-specific acid phosphatase activity was expressed constitutively in these soils. A sequential solid-phase extraction scheme and X-ray absorption spectroscopy measurements were combined to demonstrate that U(VI) was primarily precipitated as uranyl phosphate minerals at low pH, whereas it was mainly adsorbed to iron oxides and partially precipitated as uranyl phosphate at circumneutral pH. These findings suggest that, in the presence of organophosphates, microbial phosphatase activity can contribute to uranium immobilization in both low and circumneutral pH soils through the formation of stable uranyl phosphate minerals.

  9. Adsorption of divalent lead ions by zeolites and activated carbon: effects of pH, temperature, and ionic strength.

    PubMed

    Payne, Kelly B; Abdel-Fattah, Tarek M

    2004-01-01

    Lead alloy bullets used at the 2600 military small arm ranges and 9000 nonmilitary outdoor shooting ranges in the United States are a source of mobilized lead ions under conditions of low pH, significant changes in ionic strength, changes in the reduction oxidation potential (redox), and through binding metal ions to soil organic matter. Once mobile, these lead ions can contaminate adjacent soil and water. Batch adsorption kinetic and isotherm studies were conducted to compare and evaluate different types of adsorbents for lead ion removal from aqueous media. The effects on lead ion absorption from pH changes, competing ions, and temperature increases were also investigated. Adsorbent materials such as activated carbon and naturally occurring zeolites (clinoptilolite and chabazite) were selected because of their relative low cost and because the zeolites are potential point-of-use materials for mitigating wastewater runoff. Molecular sieves, Faujasite (13X) and Linde type A (5A) were selected because they provide a basis for comparison with previous studies and represent well-characterized materials. The relative rate for lead ion adsorption was: 13X > chabazite > clinoptilolite > 5A > activated carbon. Modeling lead ion adsorption by these adsorbents using the Langmuir and Freundlich isotherm expressions determined the adsorbents' capacity for lead ion removal from aqueous media. 13X, 5A, and activated carbon best fit the Langmuir isotherm expression; chabazite and clinoptilolite best fit the Freundlich isotherm. Applications of chabazite would require pH values between 4 and 11, clinoptilolite between 3 and 11, while activated carbon would operate at a pH above 7. Ionic competition reduced lead ion removal by the zeolites, but enhanced activated carbon performance. Increasing temperature improved adsorption performance for the zeolites; activated carbon lead ion adsorption was temperature independent.

  10. Possible association of Neu2 with plasma membrane fraction from mouse thymus exhibited sialidase activity with fetuin at pH 7.0 but not at pH 4.5.

    PubMed

    Kijimoto-Ochiai, Shigeko; Doi, Naoko; Fujii, Miwako; Go, Shinji; Kabayama, Kazuya; Moriya, Setsuko; Miyagi, Taeko; Koda, Toshiaki

    2013-08-01

    Compared to other organs, the mouse thymus exhibits a high level of sialidase activity in both the soluble and crude membrane fractions, as measured at neutral pH using 4MU-Neu5Ac as a substrate. The main purpose of the present study was to identify the sialidase with a high level of the activity at neutral pH in the crude membrane. Several parameters were analyzed using the soluble (S) fraction, N and D fractions that were obtained by NP-40 or DOC/NP-40 solubilization from the thymus crude membrane. The main sialidase activity in the N fraction exhibited almost the same pI as that of soluble Neu2 and 60% of the activity was removed from the membrane by three washes with 10 mM Tris-buffer, at pH 7.0. The N fraction preferentially hydrolyzed the sialic acid bond of glycoprotein and exhibited sialidase activity with fetuin at pH 7.0 but not at pH 4.5. The same activity was observed in a plasma membrane-rich fraction. To date, the removal of sialic acid from fetuin at pH 7.0 was reported only with soluble Neu2 and the membrane fraction from Neu2-transfected COS cells. We analyzed the gene that controls the sialidase activity in the crude membrane fraction at pH 7.0 using SMXA recombinant mice and found that compared with other three genes, Neu2 presented the best correlation with the activity level. We suggest that Neu2 is most likely responsible for the main activity in the N fraction, due to its association with the membrane by an unknown mechanism.

  11. Ruthenium dihydroxybipyridine complexes are tumor activated prodrugs due to low pH and blue light induced ligand release.

    PubMed

    Hufziger, Kyle T; Thowfeik, Fathima Shazna; Charboneau, David J; Nieto, Ismael; Dougherty, William G; Kassel, W Scott; Dudley, Timothy J; Merino, Edward J; Papish, Elizabeth T; Paul, Jared J

    2014-01-01

    Ruthenium drugs are potent anti-cancer agents, but inducing drug selectivity and enhancing their modest activity remain challenging. Slow Ru ligand loss limits the formation of free sites and subsequent binding to DNA base pairs. Herein, we designed a ligand that rapidly dissociates upon irradiation at low pH. Activation at low pH can lead to cancer selectivity, since many cancer cells have higher metabolism (and thus lower pH) than non-cancerous cells. We have used the pH sensitive ligand, 6,6'-dihydroxy-2,2'-bipyridine (66'bpy(OH)2), to generate [Ru(bpy)2(66'(bpy(OH)2)](2+), which contains two acidic hydroxyl groups with pKa1=5.26 and pKa2=7.27. Irradiation when protonated leads to photo-dissociation of the 66'bpy(OH)2 ligand. An in-depth study of the structural and electronic properties of the complex was carried out using X-ray crystallography, electrochemistry, UV/visible spectroscopy, and computational techniques. Notably, RuN bond lengths in the 66'bpy(OH)2 complex are longer (by ~0.3Å) than in polypyridyl complexes that lack 6 and 6' substitution. Thus, the longer bond length predisposes the complex for photo-dissociation and leads to the anti-cancer activity. When the complex is deprotonated, the 66'bpy(O(-))2 ligand molecular orbitals mix heavily with the ruthenium orbitals, making new mixed metal-ligand orbitals that lead to a higher bond order. We investigated the anti-cancer activities of [Ru(bpy)2(66'(bpy(OH)2)](2+), [Ru(bpy)2(44'(bpy(OH)2)](2+), and [Ru(bpy)3](2+) (44'(bpy(OH)2=4,4'-dihydroxy-2,2'-bipyridine) in HeLa cells, which have a relatively low pH. It is found that [Ru(bpy)2(66'(bpy(OH)2)](2+) is more cytotoxic than the other ruthenium complexes studied. Thus, we have identified a pH sensitive ruthenium scaffold that can be exploited for photo-induced anti-cancer activity.

  12. pH control of the structure, composition, and catalytic activity of sulfated zirconia

    NASA Astrophysics Data System (ADS)

    Ivanov, Vladimir K.; Baranchikov, Alexander Ye.; Kopitsa, Gennady P.; Lermontov, Sergey A.; Yurkova, Lyudmila L.; Gubanova, Nadezhda N.; Ivanova, Olga S.; Lermontov, Anatoly S.; Rumyantseva, Marina N.; Vasilyeva, Larisa P.; Sharp, Melissa; Pranzas, P. Klaus; Tretyakov, Yuri D.

    2013-02-01

    We report a detailed study of structural and chemical transformations of amorphous hydrous zirconia into sulfated zirconia-based superacid catalysts. Precipitation pH is shown to be the key factor governing structure, composition and properties of amorphous sulfated zirconia gels and nanocrystalline sulfated zirconia. Increase in precipitation pH leads to substantial increase of surface fractal dimension (up to ˜2.7) of amorphous sulfated zirconia gels, and consequently to increase in specific surface area (up to ˜80 m2/g) and simultaneously to decrease in sulfate content and total acidity of zirconia catalysts. Complete conversion of hexene-1 over as synthesized sulfated zirconia catalysts was observed even under ambient conditions.

  13. Characterization of nitrazine yellow as a photoacoustically active pH reporter molecule.

    PubMed

    Brown, Jordan E; Diaz, Lilibet; Christoff-Tempesta, Ty; Nesbitt, Kathryn M; Reed-Betts, Julia; Sanchez, John; Davies, Kevin W

    2015-04-07

    Throughout the fields of biomedical imaging, materials analysis, and routine chemical analysis, it is desirable to have a toolkit of molecules that can allow noninvasive/remote chemical sensing with minimal sample preparation. Here, we describe the photophysical properties involved in photoacoustic (PA) measurements and present a detailed analysis of the requirements and complications involved in PA sensing. We report the use of nitrazine yellow (NY) as a well-behaved PA pH reporter molecule. Both the basic and acidic forms of NY are photoacoustically well-behaved and allow for rapid and noninvasive measurement of pH in either transparent or turbid media. We also find that the serum protein-bound form of NY is photoacoustically well-behaved and should permit applications in noninvasive 3D imaging (e.g., the lymphatic system).

  14. The combined effect of pasteurization intensity, water activity, pH and incubation temperature on the survival and outgrowth of spores of Bacillus cereus and Bacillus pumilus in artificial media and food products.

    PubMed

    Samapundo, S; Heyndrickx, M; Xhaferi, R; de Baenst, I; Devlieghere, F

    2014-07-02

    The objective of the study was to evaluate the combined effects of pasteurization intensity (no heat treatment and 10 min at 70, 80 and 90 °C), water activity (aw) (0.960-0.990), pH (5.5-7.0) and storage temperature (7 and 10 °C) on the survival and outgrowth of psychrotolerant spores of Bacillus cereus FF119b and Bacillus pumilus FF128a. The experiments were performed in both artificial media and a validation was performed on real food products (cream, béchamel sauce and mixed vegetable soup). It was determined that in general, heat treatments of 10 min at 70 °C or 80 °C activated the spores of both B. cereus FF119b and B. pumilus FF128a, resulting in faster outgrowth compared to native (non-heat treated) spores. A pasteurization treatment of 10 min at 90 °C generally resulted in the longest lag periods before outgrowth of both isolates. Some of the spores were inactivated by this heat treatment, with more inactivation being observed the lower the pH value of the heating medium. Despite this, it was also observed that under some conditions the remaining (surviving) spores were actually activated as their outgrowth took place after a shorter period of time compared to native non-heated spores. While the response of B. cereus FF119b to the pasteurization intensity in cream and béchamel sauce was similar to the trends observed in the artificial media at 10 °C, in difference, outgrowth was only observed at 7 °C in both products when the spores had been heated for 10 min at 80 °C. Moreover, no inactivation was observed in cream or béchamel sauce when the spores were heated for 10 min at 90 °C in these two products. This was attributed to the protective effect of fat in the cream and the ingredients in the béchamel sauce. The study provides some insight into the potential microbial (stability and safety) consequences of the current trend towards milder heat treatments which is being pursued in the food industry.

  15. [Effects of abiotic factors (temperature, pH, heavy metals) on activities of glycosidases in invertebrate animals].

    PubMed

    Golovanova, I L

    2011-01-01

    We have established differences in amylolytical activity in the whole organisms of several invertebrate animals (larvae of chironomids Chironomnus plumosus, pond snail Limnaea stagnalis, orb snail Planorbis corneus, faucet snail Bithynia tentaculata, and zebra mussel Dreissena polymorpha)--potential objects of nutrition of fish benthophages under separate and combined actions of temperature (0, 10 or 20 degrees C), pH (5.0, 7.4 or 8.3), and ions of heavy metals (copper, zinc at a concentration of 10 mg/l) in vitro. The strongest inhibitory effect on the amylolytical activity in tissues of the studied animals was produced by a combination of the low temperature, acidic pH values, and the presence of the heavy metal ions. Enzymes of chironomids were more resistant to action of the studied agents as compared with mollusks.

  16. Sewage sludge pretreatment by microwave irradiation combined with activated carbon fibre at alkaline pH for anaerobic digestion.

    PubMed

    Sun, Dedong; Guo, Sixiao; Ma, Nina; Wang, Guowen; Ma, Chun; Hao, Jun; Xue, Mang; Zhang, Xinxin

    2016-01-01

    This research focuses on the effects of microwave-assisted activated carbon fibre (ACF) (MW-ACF) treatment on sewage sludge at alkaline pH. The disintegration and biodegradability of sewage sludge were studied. It was found that the MW-ACF process at alkaline pH provided a rapid and efficient process to disrupt the microbial cells in the sludge. The results suggested that when irradiated at 800 W MW for 110 s with a dose of 1.0 g ACF/g solid concentration (SS) at pH 10.5, the MW-ACF pretreatment achieved 55% SS disintegration, 23% greater than the value of MW alone (32%). The concentration of total nitrogen, total phosphorus, supernatant soluble chemical oxygen demand, protein, and polysaccharide increased by 60%, 144%, 145%, 74%, and 77%, respectively. An increase in biogas production by 63.7% was achieved after 20 days of anaerobic digestion (AD), compared to the control. The results indicated that the MW-ACF pretreatment process at alkaline pH provides novel sludge management options in disintegration of sewage sludge for further AD.

  17. Activation of AMP-activated protein kinase regulates hippocampal neuronal pH by recruiting Na(+)/H(+) exchanger NHE5 to the cell surface.

    PubMed

    Jinadasa, Tushare; Szabó, Elöd Z; Numat, Masayuki; Orlowski, John

    2014-07-25

    Strict regulation of intra- and extracellular pH is an important determinant of nervous system function as many voltage-, ligand-, and H(+)-gated cationic channels are exquisitely sensitive to transient fluctuations in pH elicited by neural activity and pathophysiologic events such as hypoxia-ischemia and seizures. Multiple Na(+)/H(+) exchangers (NHEs) are implicated in maintenance of neural pH homeostasis. However, aside from the ubiquitous NHE1 isoform, their relative contributions are poorly understood. NHE5 is of particular interest as it is preferentially expressed in brain relative to other tissues. In hippocampal neurons, NHE5 regulates steady-state cytoplasmic pH, but intriguingly the bulk of the transporter is stored in intracellular vesicles. Here, we show that NHE5 is a direct target for phosphorylation by the AMP-activated protein kinase (AMPK), a key sensor and regulator of cellular energy homeostasis in response to metabolic stresses. In NHE5-transfected non-neuronal cells, activation of AMPK by the AMP mimetic AICAR or by antimycin A, which blocks aerobic respiration and causes acidification, increased cell surface accumulation and activity of NHE5, and elevated intracellular pH. These effects were effectively blocked by the AMPK antagonist compound C, the NHE inhibitor HOE694, and mutation of a predicted AMPK recognition motif in the NHE5 C terminus. This regulatory pathway was also functional in primary hippocampal neurons, where AMPK activation of NHE5 protected the cells from sustained antimycin A-induced acidification. These data reveal a unique role for AMPK and NHE5 in regulating the pH homeostasis of hippocampal neurons during metabolic stress.

  18. Activation of AMP-activated Protein Kinase Regulates Hippocampal Neuronal pH by Recruiting Na+/H+ Exchanger NHE5 to the Cell Surface*

    PubMed Central

    Jinadasa, Tushare; Szabó, Elöd Z.; Numata, Masayuki; Orlowski, John

    2014-01-01

    Strict regulation of intra- and extracellular pH is an important determinant of nervous system function as many voltage-, ligand-, and H+-gated cationic channels are exquisitely sensitive to transient fluctuations in pH elicited by neural activity and pathophysiologic events such as hypoxia-ischemia and seizures. Multiple Na+/H+ exchangers (NHEs) are implicated in maintenance of neural pH homeostasis. However, aside from the ubiquitous NHE1 isoform, their relative contributions are poorly understood. NHE5 is of particular interest as it is preferentially expressed in brain relative to other tissues. In hippocampal neurons, NHE5 regulates steady-state cytoplasmic pH, but intriguingly the bulk of the transporter is stored in intracellular vesicles. Here, we show that NHE5 is a direct target for phosphorylation by the AMP-activated protein kinase (AMPK), a key sensor and regulator of cellular energy homeostasis in response to metabolic stresses. In NHE5-transfected non-neuronal cells, activation of AMPK by the AMP mimetic AICAR or by antimycin A, which blocks aerobic respiration and causes acidification, increased cell surface accumulation and activity of NHE5, and elevated intracellular pH. These effects were effectively blocked by the AMPK antagonist compound C, the NHE inhibitor HOE694, and mutation of a predicted AMPK recognition motif in the NHE5 C terminus. This regulatory pathway was also functional in primary hippocampal neurons, where AMPK activation of NHE5 protected the cells from sustained antimycin A-induced acidification. These data reveal a unique role for AMPK and NHE5 in regulating the pH homeostasis of hippocampal neurons during metabolic stress. PMID:24936055

  19. Simultaneous spatiotemporal mapping of in situ pH and bacterial activity within an intact 3D microcolony structure

    NASA Astrophysics Data System (ADS)

    Hwang, Geelsu; Liu, Yuan; Kim, Dongyeop; Sun, Victor; Aviles-Reyes, Alejandro; Kajfasz, Jessica K.; Lemos, Jose A.; Koo, Hyun

    2016-09-01

    Biofilms are comprised of bacterial-clusters (microcolonies) enmeshed in an extracellular matrix. Streptococcus mutans can produce exopolysaccharides (EPS)-matrix and assemble microcolonies with acidic microenvironments that can cause tooth-decay despite the surrounding neutral-pH found in oral cavity. How the matrix influences the pH and bacterial activity locally remains unclear. Here, we simultaneously analyzed in situ pH and gene expression within intact biofilms and measured the impact of damage to the surrounding EPS-matrix. The spatiotemporal changes of these properties were characterized at a single-microcolony level following incubation in neutral-pH buffer. The middle and bottom-regions as well as inner-section within the microcolony 3D structure were resistant to neutralization (vs. upper and peripheral-region), forming an acidic core. Concomitantly, we used a green fluorescent protein (GFP) reporter to monitor expression of the pH-responsive atpB (PatpB::gfp) by S. mutans within microcolonies. The atpB expression was induced in the acidic core, but sharply decreased at peripheral/upper microcolony regions, congruent with local pH microenvironment. Enzymatic digestion of the surrounding matrix resulted in nearly complete neutralization of microcolony interior and down-regulation of atpB. Altogether, our data reveal that biofilm matrix facilitates formation of an acidic core within microcolonies which in turn activates S. mutans acid-stress response, mediating both the local environment and bacterial activity in situ.

  20. Simultaneous spatiotemporal mapping of in situ pH and bacterial activity within an intact 3D microcolony structure

    PubMed Central

    Hwang, Geelsu; Liu, Yuan; Kim, Dongyeop; Sun, Victor; Aviles-Reyes, Alejandro; Kajfasz, Jessica K.; Lemos, Jose A.; Koo, Hyun

    2016-01-01

    Biofilms are comprised of bacterial-clusters (microcolonies) enmeshed in an extracellular matrix. Streptococcus mutans can produce exopolysaccharides (EPS)-matrix and assemble microcolonies with acidic microenvironments that can cause tooth-decay despite the surrounding neutral-pH found in oral cavity. How the matrix influences the pH and bacterial activity locally remains unclear. Here, we simultaneously analyzed in situ pH and gene expression within intact biofilms and measured the impact of damage to the surrounding EPS-matrix. The spatiotemporal changes of these properties were characterized at a single-microcolony level following incubation in neutral-pH buffer. The middle and bottom-regions as well as inner-section within the microcolony 3D structure were resistant to neutralization (vs. upper and peripheral-region), forming an acidic core. Concomitantly, we used a green fluorescent protein (GFP) reporter to monitor expression of the pH-responsive atpB (PatpB::gfp) by S. mutans within microcolonies. The atpB expression was induced in the acidic core, but sharply decreased at peripheral/upper microcolony regions, congruent with local pH microenvironment. Enzymatic digestion of the surrounding matrix resulted in nearly complete neutralization of microcolony interior and down-regulation of atpB. Altogether, our data reveal that biofilm matrix facilitates formation of an acidic core within microcolonies which in turn activates S. mutans acid-stress response, mediating both the local environment and bacterial activity in situ. PMID:27604325

  1. Effect of pH and sodium chloride on growth of Bacillus cereus in laboratory media and certain foods.

    PubMed

    Raevuori, M; Genigeorgis, C

    1975-01-01

    The effects of NaCl concentration, pH, and water activity (aw) on the ability of vegetative cells of Bacillus cereus to initiate aerobic growth in brain heart infusion broth at 30 C were studied in a factorial design experiment. By using multiple regression techniques, equations were derived which related the decimal reduction of the bacterial population to the concentration of NaCl and pH of broth to which the population was exposed. From these equations, the percentage of inoculated cells capable of initiating growth could be calculated. The reliability of these equations in foods was tested in laboratory-processed meat and rice media. The foods were less inhibitory than the broths, so that accurate prediction of growth initiation in foods was not possible by using the developed formulas. The impact of this type of quantitative study on the development of specific microbial standards for foods is discussed. When the NaCl concentration is increased, the aw is decreased and, with increased deviation of pH from optimum, more concentrated inoculum of B. cereus cells is needed to assure initiation of growth in culture media and foods.

  2. Effect of pH and Sodium Chloride on Growth of Bacillus cereus in Laboratory Media and Certain Foods

    PubMed Central

    Raevuori, Marrku; Genigeorgis, Constantin

    1975-01-01

    The effects of NaCl concentration, pH, and water activity (aw) on the ability of vegetative cells of Bacillus cereus to initiate aerobic growth in brain heart infusion broth at 30 C were studied in a factorial design experiment. By using multiple regression techniques, equations were derived which related the decimal reduction of the bacterial population to the concentration of NaCl and pH of broth to which the population was exposed. From these equations, the percentage of inoculated cells capable of initiating growth could be calculated. The reliability of these equations in foods was tested in laboratory-processed meat and rice media. The foods were less inhibitory than the broths, so that accurate prediction of growth initiation in foods was not possible by using the developed formulas. The impact of this type of quantitative study on the development of specific microbial standards for foods is discussed. When the NaCl concentration is increased, the aw is decreased and, with increased deviation of pH from optimum, more concentrated inoculum of B. cereus cells is needed to assure initiation of growth in culture media and foods. PMID:234158

  3. Atypical presentation of axillary web syndrome (AWS) in a male squash player: a case report

    PubMed Central

    Welsh, Patrick; Gryfe, David

    2016-01-01

    Axillary Web Syndrome (AWS), also known as lymphatic cording, refers to a condition in which a rope-like soft-tissue density develops in the axilla. It usually appears in the 5 to 8 week period following breast cancer surgery and can lead to shoulder pain and restricted motion. We present a case of AWS in a male squash player with no history of breast cancer or surgery following a period of intense exercise. This case highlights the rare presentation of AWS in a male patient and raises awareness for the health care practitioner who may not suspect this condition in this population. PMID:28065989

  4. Exploring the active site of tripeptidyl-peptidase II through studies of pH dependence of reaction kinetics.

    PubMed

    Eklund, Sandra; Lindås, Ann-Christin; Hamnevik, Emil; Widersten, Mikael; Tomkinson, Birgitta

    2012-04-01

    Tripeptidyl-peptidase II (TPP II) is a subtilisin-like serine protease which forms a large enzyme complex (>4MDa). It is considered a potential drug target due to its involvement in specific physiological processes. However, information is scarce concerning the kinetic characteristics of TPP II and its active site features, which are important for design of efficient inhibitors. To amend this, we probed the active site by determining the pH dependence of TPP II catalysis. Access to pure enzyme is a prerequisite for kinetic investigations and herein we introduce the first efficient purification system for heterologously expressed mammalian TPP II. The pH dependence of kinetic parameters for hydrolysis of two different chromogenic substrates, Ala-Ala-Phe-pNA and Ala-Ala-Ala-pNA, was determined for murine, human and Drosophila melanogaster TPP II as well as mutant variants thereof. The investigation demonstrated that TPP II, in contrast to subtilisin, has a bell-shaped pH dependence of k(cat)(app)/K(M) probably due to deprotonation of the N-terminal amino group of the substrate at higher pH. Since both the K(M) and k(cat)(app) are lower for cleavage of AAA-pNA than for AAF-pNA we propose that the former can bind non-productively to the active site of the enzyme, a phenomenon previously observed with some substrates for subtilisin. Two mutant variants, H267A and D387G, showed bell-shaped pH-dependence of k(cat)(app), possibly due to an impaired protonation of the leaving group. This work reveals previously unknown differences between TPP II orthologues and subtilisin as well as features that might be conserved within the entire family of subtilisin-like serine peptidases.

  5. Activation of immunity, immune response, antioxidant ability, and resistance against Vibrio alginolyticus in white shrimp Litopenaeus vannamei decrease under long-term culture at low pH.

    PubMed

    Chen, Yu-Yuan; Chen, Jiann-Chu; Tseng, Kuei-Chi; Lin, Yong-Chin; Huang, Chien-Lun

    2015-10-01

    The growth, activation of immunity, immune parameters, and transcript levels of cytMnSOD, mtMnSOD, ecCuZnSOD, glutathione peroxidase (GPx), catalase, lysozyme, and penaeidin 3a were examined in white shrimp Litopenaeus vannamei reared at pH 6.8 and 8.1 after 24 weeks. No significant difference in growth was observed between the two groups. An in vitro study indicated that phenoloxidase activity and respiratory bursts (RB, release of the superoxide anion) were significantly higher in the haemocytes of pH 8.1 shrimp (shrimp reared at pH 8.1) than in pH 6.8 shrimp (shrimp reared at pH 6.8). An in vivo study indicated that the levels of immune parameters of pH 8.1 shrimp were significantly higher than in pH 6.8 shrimp, and the transcript levels of cytMnSOD, ecCuZnSOD, glutathione peroxidase, lysozyme, and penaeidin 3a were down-regulated in pH 6.8 shrimp. In another experiment, shrimp reared at pH 6.8 and 8.1 for 24 weeks were challenged with Vibrio alginolyticus. The mortality rate of pH 6.8 shrimp was significantly higher than in pH 8.1 shrimp over 12-168 h. Phagocytic activity, phagocytic index, and clearance efficiency to V. alginolyticus were significantly lower in pH 6.8 shrimp. We concluded that shrimp under long-term culture at pH 6.8 exhibited decreased resistance against V. alginolyticus as evidenced by reductions in the activation of immunity and immune parameters together with decreased transcript levels of cytMnSOD, ecCuZnSOD, GPx, lysozyme, and penaeidin 3a.

  6. LEGUS Discovery of a Light Echo Around Supernova 2012aw

    NASA Astrophysics Data System (ADS)

    Van Dyk, Schuyler D.; Lee, Janice C.; Anderson, Jay; Andrews, Jennifer E.; Calzetti, Daniela; Bright, Stacey N.; Ubeda, Leonardo; Smith, Linda J.; Sabbi, Elena; Grebel, Eva K.; Herrero, Artemio; de Mink, Selma E.

    2015-06-01

    We have discovered a luminous light echo around the normal Type II-Plateau Supernova (SN) 2012aw in Messier 95 (M95; NGC 3351), detected in images obtained approximately two years after explosion with the Wide Field Channel 3 on board the Hubble Space Telescope by the Legacy ExtraGalactic Ultraviolet Survey. The multi-band observations span from the near-ultraviolet through the optical (F275W, F336W, F438W, F555W, and F814W). The apparent brightness of the echo at the time was ˜21-22 mag in all of these bands. The echo appears circular, although less obviously as a ring, with an inhomogeneous surface brightness, in particular, a prominent enhanced brightness to the southeast. The SN itself was still detectable, particularly in the redder bands. We are able to model the light echo as the time-integrated SN light scattered off of diffuse interstellar dust in the SN environment. We have assumed that this dust is analogous to that in the Milky Way with {R}V=3.1. The SN light curves that we consider also include models of the unobserved early burst of light from the SN shock breakout. Our analysis of the echo suggests that the distance from the SN to the scattering dust elements along the echo is ≈ 45 pc. The implied visual extinction for the echo-producing dust is consistent with estimates made previously from the SN itself. Finally, our estimate of the SN brightness in F814W is fainter than that measured for the red supergiant star at the precise SN location in pre-SN images, possibly indicating that the star has vanished and confirming it as the likely SN progenitor.

  7. 47 CFR 27.1160 - Cost-sharing requirements for AWS.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... entities that benefit from the clearance of this spectrum by other AWS entities or by a voluntarily... spectrum leasing arrangement (as set forth in part 1, subpart X of this chapter) and the spectrum...

  8. Chewing activity, saliva production, and ruminal pH of primiparous and multiparous lactating dairy cows.

    PubMed

    Maekawa, M; Beauchemin, K A; Christensen, D A

    2002-05-01

    Four multiparous (MP) and four primiparous (PP) ruminally cannulated lactating Holstein cows were used in a double 4 x 4 Latin square design to study the chewing behavior, saliva production, and ruminal pH of cows in the first or subsequent lactation. Cows were fed one of four diets; three total mixed rations containing 40, 50, or 60% silage (DM basis), and a separate ingredient diet containing 50% concentrate. Dry matter intake was higher for MP cows than for PP cows (19.2 vs. 17.1 kg/d) but not as a percentage of body weight (2.97 +/- 0.06%). Multiparous cows spent more time eating than PP cows (260 vs. 213 min/d, respectively), even after adjustment for dry matter intake (13.8 vs. 12.4 min/kg DM). Multiparous cows also spent more time ruminating per day than PP cows (560 vs. 508 min/d, respectively). Eating salivation rate was not affected by parity, but resting salivation rate was higher for MP cows than for PP. Although MP cows spent more time chewing than PP cows, total daily saliva production was only numerically higher for MP cows because the increase in saliva produced during chewing was accompanied by a decrease in saliva produced during resting. Furthermore, pH profiles tended to be lower for MP cows than for PP cows. Multiparous cows may have a greater risk of incurring acidosis than PP cows because increased salivary secretion associated with increased chewing may not sufficiently compensate the increment of fermentation acids produced in the rumen due to high feed intake.

  9. Effect of pH on activities of novel beta-lactamases and beta-lactamase inhibitors against these beta-lactamases.

    PubMed Central

    Ohsuka, S; Arakawa, Y; Horii, T; Ito, H; Ohta, M

    1995-01-01

    The effects of acidic conditions on activities of seven beta-lactamases--TEM-1 (class A), KOXY (class A), IMP-1 (class B), AmpC (class C), MOX-1 (class C), OXA-5 (class D), and PSE-2 (class D)--and their inhibitors were measured. The enzymatic activities of KOXY, IMP-1, and MOX-1 at pH 5.8 were slightly lower than those at pH 7.5. However, the activities of PSE-2 and OXA-5 were greatly reduced at pH 5.8. All of the beta-lactamase inhibitors tested had poorer inhibitory activities at pH 5.8 than at pH 7.5 except clavulanic acid for TEM-1. PMID:7486932

  10. LESSONS LEARNED, HEADQUARTERS, 4TH BATTALION (AW)(SP), 60TH ARTILLERY

    DTIC Science & Technology

    The 4th Battalion (AW)(SP) 60th Artillery with attached Battery E (MG), 41st Artillery, remained assigned to I Field Force Vietnam, attached to I ...Battalion (AW)(SP), 60th Artillery, with attached Battery E (MG), 41st Artillery, was detached from 41st Artillery Group and fully attached to I Field Force...States and Free World Military Assistance Forces throughout the II Corps Tactical Zone and the I Corps Tactical Zone.

  11. On the pH dependent behavior of the firefly bioluminescence: protein dynamics and water content in the active pocket.

    PubMed

    Kim, Hyun Woo; Rhee, Young Min

    2013-06-20

    Understanding bioluminescence presents fascinating challenges for fundamental sciences and numerous opportunities for practical applications. As a representative example, the firefly bioluminescent system has been intensively studied in both experimental and computational areas. However, there are still remaining questions regarding especially the detailed protein dynamics and the mechanisms of its color modulation. Here, we report on the pH dependent behavior of the firefly bioluminescence primarily based on molecular dynamics simulations. We find that the overall protein structure is generally resilient to pH variations. As the protein does not exhibit any structural distortions that can affect the emission property, we next focus on the dynamics in the active pocket and its effect on color modulation by adopting different protonation states in the pocket. With this, we observe red-shifted emissions at acidic conditions as consistent with previous studies. Most importantly, we find that a water molecule in the active pocket can mediate flexible motions of neighboring groups, which can subsequently modify the emission properties to a substantial degree. Based on the observations, we propose that the active pocket is in a dry condition during the luminescence process. Our results highlight the importance of understanding the role of the dynamics near the active pocket in modulating bioluminescence.

  12. Design of a Water Soluble Fluorescent 3-Hydroxy-4-Pyridinone Ligand Active at Physiological pH Values.

    PubMed

    Leite, Andreia; Silva, Ana M G; Coutinho, Catarina; Cunha-Silva, Luís; de Castro, Baltazar; Rangel, Maria

    2016-09-01

    In the present work we report the structure and the spectroscopic characterization of a new fluorescent 3-hydroxy-4-pyridinone ligand D-3,4-HPO. The synthesis of the compound was performed in two steps, which involve the reaction of the commercially available fluorophore dansyl chloride with a 3-hydroxy-4-pyridinone chelating unit and further deprotection. The new fluorescent chelator was characterized in the solid state by single-crystal X-ray diffraction and in solution by NMR, MS, absorption and fluorescence spectroscopies. The analysis of the variation of the absorption spectrum with pH allowed the determination of four pK a values (pK a1  = 3.50, pK a2  = 4.50, pK a3  = 9.60, pK a4  = 10.20) and establishment of the corresponding distribution diagram. The study of the fluorescence properties of the ligand show that in the pH range between 4 and 9 the fluorescence intensity is constant and has its maximum value thus allowing its further use at physiological pH values. The interaction of the ligand with copper(II) was accessed by fluorescence spectroscopy in MOPS buffer and the results show that the presence of copper(II) quenches the fluorescence of the ligand in ca 94 % at a ligand: metal ratio of 2:1. The latter result is consistent with the formation of a copper(II) complex with the bidentate ligand, as confirmed by the EPR spectroscopy. Graphical Abstract New water soluble fluorescent ligand active at physiological pH values.

  13. Testing the limits of rational design by engineering pH sensitivity into membrane-active peptides.

    PubMed

    Wiedman, Gregory; Wimley, William C; Hristova, Kalina

    2015-04-01

    In this work, we sought to rationally design membrane-active peptides that are triggered by low pH to form macromolecular-sized pores in lipid bilayers. Such peptides could have broad utility in biotechnology and in nanomedicine as cancer therapeutics or drug delivery vehicles that promote release of macromolecules from endosomes. Our approach to rational design was to combine the properties of a pH-independent peptide, MelP5, which forms large pores allowing passage of macromolecules, with the properties of two pH-dependent membrane-active peptides, pHlip and GALA. We created two hybrid sequences, MelP5_Δ4 and MelP5_Δ6, by using the distribution of acidic residues on pHlip and GALA as a guide to insert acidic amino acids into the amphipathic helix of MelP5. We show that the new peptides bind to lipid bilayers and acquire secondary structure in a pH-dependent manner. The peptides also destabilize bilayers in a pH-dependent manner, such that lipid vesicles release the small molecules ANTS/DPX at low pH only. Thus, we were successful in designing pH-triggered pore-forming peptides. However, no macromolecular release was observed under any conditions. Therefore, we abolished the unique macromolecular poration properties of MelP5 by introducing pH sensitivity into its sequence. We conclude that the properties of pHlip, GALA, and MelP5 are additive, but only partially so. We propose that this lack of additivity is a limitation in the rational design of novel membrane-active peptides, and that high-throughput approaches to discovery will be critical for continued progress in the field.

  14. Modeling growth of three bakery product spoilage molds as a function of water activity, temperature and pH.

    PubMed

    Dagnas, Stéphane; Onno, Bernard; Membré, Jeanne-Marie

    2014-09-01

    The objective of this study was to quantify the effect of water activity, pH and storage temperature on the growth of Eurotium repens, Aspergillus niger and Penicillium corylophilum, isolated from spoiled bakery products. Moreover, the behaviors of these three mold species were compared to assess whether a general modeling framework may be set and re-used in future research on bakery spoilage molds. The mold growth was modeled by building two distinct Gamma-type secondary models: one on the lag time for growth and another one on the radial growth rate. A set of 428 experimental growth curves was generated. The effect of temperature (15-35 °C), water activity (0.80-0.98) and pH (3-7) was assessed. Results showed that it was not possible to apply the same set of secondary model equations to the three mold species given that the growth rate varied significantly with the factors pH and water activity. In contrast, the temperature effect on both growth rate and lag time of the three mold species was described by the same equation. The equation structure and model parameter values of the Gamma models were also compared per mold species to assess whether a relationship between lag time and growth rate existed. There was no correlation between the two growth responses for E. repens, but a slight one for A. niger and P. corylophilum. These findings will help in determining bakery product shelf-life and guiding future work in the predictive mycology field.

  15. Growth of and fumitremorgin production by Neosartorya fischeri as affected by temperature, light, and water activity.

    PubMed Central

    Nielsen, P V; Beuchat, L R; Frisvad, J C

    1988-01-01

    The effects of temperature, light, and water activity (aw) on the growth and fumitremorgin production of a heat-resistant mold, Neosartorya fischeri, cultured on Czapek Yeast Autolysate agar (CYA) were studied for incubation periods of up to 74 days. Colonies were examined visually, and extracts of mycelia and CYA on which the mold was cultured were analyzed for mycotoxin content by high-performance liquid chromatography. Growth always resulted in the production of the tremorgenic mycotoxins verruculogen and fumitremorgins A and C. The optimum temperatures for the production of verruculogen and fumitremorgins A and C on CYA at pH 7.0 were 25, 30, and 37 degrees C, respectively. The production of fumitremorgin C by N. fischeri has not been previously reported. Fumitremorgin production was retarded at 15 degrees C, but an extension of the incubation period resulted in concentrations approaching those observed at 25 degrees C. Light clearly enhanced fumitremorgin production on CYA (pH 7.0, 25 degrees C), but not as dramatically as did the addition of glucose, fructose, or sucrose to CYA growth medium (pH 3.5, 25 degrees C). Growth and fumitremorgin production was greatest at aw of 0.980 on CYA supplemented with glucose or fructose and at aw of 0.990 on CYA supplemented with sucrose. Growth and fumitremorgin production were observed at aw as low as 0.925 on glucose-supplemented CYA but not at aw lower than 0.970 on CYA supplemented with sucrose. Verruculogen was produced in the highest amount on all test media, followed by fumitremorgin A and fumitremorgin C. PMID:3415223

  16. Rapid X-Ray Variability of Active Galaxies. Ph.D. Thesis

    NASA Technical Reports Server (NTRS)

    Tennant, A. F., Jr.

    1983-01-01

    Active galactic nuclei are luminous sources of X-rays. The thesis that the X-rays are generated within 10 gravitational radii from the central object is tested. A very sensitive search for rapid ( 1 day) X-ray variability from active galaxies was made.

  17. Mathematical modelling of the combined effect of water activity, pH and redox potential on the heat destruction.

    PubMed

    Reichart, O; Mohácsi-Farkas, C

    1994-12-01

    Heat destruction of seven foodborne microorganisms (Lactobacillus plantarum, Lactobacillus brevis, Saccharomyces cerevisiae, Zygosaccharomyces bailii, Yarrowia lipolytica, Paecilomyces varioti and Neosartoria fischeri) as a function of the temperature, pH, redox potential and water activity was studied in synthetic heating media. Several mathematical models were developed for describing the heat destruction rate, most of them resulted in a good correlation between the fitted and measured values. The determination coefficients of the model-fitting were the best in case of lactobacilli and moulds (0.96-0.99) and the worst in case of the yeasts (0.81-0.88).

  18. Study of the synergistic effects of salinity, pH, and temperature on the surface-active properties of biosurfactants produced by Lactobacillus pentosus.

    PubMed

    Bello, Xanel Vecino; Devesa-Rey, Rosa; Cruz, José Manuel; Moldes, Ana Belén

    2012-02-08

    Many studies have investigated the effects of pH, temperature, and salinity on the surface-active properties of various surfactants, although in most cases the variables have been studied separately, without considering the effects of any interactions between them. In the present study, a Box-Behnken factorial design was applied to study the effects of pH, temperature, and salinity on the surface-active properties of a biosurfactant produced by Lactobacillus pentosus. The data obtained enabled development of a second-order model describing the interrelationships between operational and experimental variables, by equations including linear, interaction, and quadratic terms. The variable that had the greatest effect on the surface-active properties of the biosurfactant was pH. Moreover, at pH 3-5.5, decreases in salinity and temperature acted synergistically, reducing the surface tension of the biosurfactant; at pH 8, the same effect was observed with increasing salinity and temperature.

  19. Photometric observations and numerical modeling of AW Sge

    NASA Astrophysics Data System (ADS)

    Montgomery, M. M.; Voloshina, I.; Goel, Amit

    2016-01-01

    In this work, we present R-band photometric light curves of Cataclysmic Variable AW Sge, an SU Uma type, near superoutburst maximum. The positive superhump shape changes over three days, from single peaked on October 11, 2013 to one maximum near phase ϕ ˜ 0.3 followed by minor peaks near phases ϕ ˜ 0.6 and ϕ ˜ 0.9, respectively, on October 13, 2013. Using the maxima from October 11-13, 2013 (JD 2456577-2356579), the observed positive superhump period is 0.074293 ± 0.000025 days. In addition to the observations, we also provide a three dimensional Smoothed Particle Hydrodynamic simulation near superoutburst maximum, for comparison, assuming a secondary-to-primary mass ratio q =M2 /M1 = 0.6 M⊙/0.132 M⊙ = 0.22. The simulation produces positive superhump shapes that are similar to the observations. The simulated positive superhump has a period of 0.076923 days, which is approximately 6% longer than the orbital period, assuming an orbital period Porb = 0.0724 days. The 3.5% difference from the observed positive superhump period is likely due to the assumptions used in generating the simulations, as the orbital period and masses are not well known. From an analysis of the simulated positive superhump shape near superoutburst maximum, the maximum occurs near ϕ ˜ 0.3, when the disk is highly elliptical and eccentric and at least one of the two density waves is compressing with the disk rim. Based on the simulation, we find that the disk may be tilted and precessing in the retrograde direction at a time that is just before the next outburst and/or superoutburst.

  20. Improved activity and pH stability of E. coli ATCC 11105 penicillin acylase by error-prone PCR.

    PubMed

    Balci, Huseyin; Ozturk, Merve Tuzlakoglu; Pijning, Tjaard; Ozturk, Saliha Issever; Gumusel, Fusun

    2014-05-01

    Penicillin G acylase is the key enzyme used in the industrial production of β-lactam antibiotics. This enzyme hydrolyzes penicillin G and related β-lactam antibiotics releasing 6-aminopenicillanic acid, which is an intermediate in the production of semisynthetic penicillins. To improve the enzymatic activity of Escherichia coli penicillin acylase, sequential rounds of error-prone polymerase chain reaction were applied to the E. coli pac gene. After the second round of evolution, the best mutant M2234 with enhanced activity was selected and analyzed. DNA sequence analyses of M2234 revealed that one amino acid residue (K297I), located far from the center of the catalytic pocket, was changed. This mutant (M2234) has a specific activity 4.0 times higher than the parent enzyme and also displayed higher stability at pH 10.

  1. In vivo selection for the enhancement of Thermotoga maritima exopolygalacturonase activity at neutral pH and low temperature.

    PubMed

    Drone, Jullien; Dion, Michel; Tellier, Charles; Rabiller, Claude

    2007-01-01

    The aim of this study was to develop an Escherichia coli-based metabolic selection system for the uncovering of new oligogalacturonate-active enzymes. Based on the expression of the specific permease TogMNAB, this system enabled the entry of oligogalacturonates into the cytoplasm of E. coli thus providing a modified strain usable for this purpose. This tool was used for the metabolic selection of Thermotoga maritima exopolygalacturonase (TmGalU) mutants enabling the uptake of sodium trigalacturonate as the sole carbon source by the bacterium. In only one round of error-prone PCR and selection, mutants of TmGalU with a 4-fold increased turnover at pH 7.0 and 2-fold more active at 37 degrees C than wild-type enzyme were isolated. These results show the versatility of this strain for the evolution of oligogalacturonate-active enzymes.

  2. Inter and intrasite analyses of cultural materials from U20aw, Nye County, Nevada

    SciTech Connect

    Hicks, P.A.; Pippin, L.C.; Henton, G.H.

    1991-12-01

    This archaeological presents the results of the analyses of the data derived from all sites investigated during the data recovery operations on Drill Hole U20aw on the Nevada Test Site, Nye County Nevada. These sites were determined to be potentially eligible for inclusion in the national Register of Historic Places. Detailed analyses were focused on the spatial distribution of artifacts and features within and between sites in the southern portion of the study area. These analyses indicate that one area served principally as a temporary camp, while the area around the canyonhead to the east seems to have been used as a site for both temporary camps and special activity loci. Projectile point styles suggest that the area was occupied from the Early Archaic into the early Historic period. Analyses of the artifacts that were recovered indicate that obsidian was the preferred material for all classes of flaked stone tools. All stages of lithic reduction are represented on the sites, but core reduction and thinning of bifaces appear to have been the primary activities. Processing of floral foods is indicated by the presence of several ground stone artifacts. Pinyon nuts and other items appear to have been stored in several areas as evidenced by the presence of several rock features that may have served as caches.

  3. Peripheral administration of low pH solutions causes activation and sensitisation of convergent dorsal horn neurones in the anaesthetised rat.

    PubMed

    Carpenter, K J; Nandi, M; Dickenson, A H

    2001-02-09

    This is the first study to examine the effects of peripheral administration of acid on the activity of dorsal horn neurones in vivo. Extracellular recordings from convergent neurones revealed increases in neuronal activity evoked by administration of low pH solutions into the peripheral receptive field. Threshold for activity ranged from pH 5.85 to 2.5. The magnitude of responses increased with decreasing pH; maximum effects were achieved with pH 2.5 (648+/-181 action potentials/60 s, as compared to control-evoked activity of 86.3+/-29 action potentials/60 s). Activity lasted for up to 60 s, likely to represent the time for which the solutions were able to surmount the buffering capacity of the intact hindpaw. Significant sensitisation of the neurones to both innocuous (von Frey filament 9 g) and noxious (30 g) mechanical punctate stimuli was also observed.

  4. Sulfoximine-mediated syntheses of optically active alcohols. Ph.D. Thesis

    NASA Technical Reports Server (NTRS)

    Stark, C. J., Jr.

    1978-01-01

    Several routes are described for the production of optically active secondary and tertiary alcohols. In all cases, the asymmetry emanates from the use of (+)-(S)-N,S-dimethyl-S-phenyl-sulfoximine (1) at some point in the variation of the diastereomers. One route relies upon the separation of the diastereomers produced from the condensation of (+)-(S)-(N-methylphenyl-sulfonimidoyl) methyllithium with prochiral aldehydes and ketones. Subsequent carbon-sulfur bond cleavage of the separated diastereomeric beta-hydroxysulfoximines yields optically active alcohols. Alternatively, beta-hydroxysulfoximines were produced from the reduction of chiral beta-ketosulfoximines. The reductions were most successfully achieved with diborane generated externally and bubbled into a toluene solution of the ketone at -78 C. Optically active alcohols were also produced from prochiral ketones by reduction with diborane or lithium aluminum hydride complexes of resolved diastereomers of beta-hydroxysulfoximines.

  5. Design for active and passive flutter suppression and gust alleviation. Ph.D. Thesis

    NASA Technical Reports Server (NTRS)

    Karpel, M.

    1981-01-01

    Analytical design techniques for active and passive control of aeroelastic systems are based on a rational approximation of the unsteady aerodynamic loads in the entire Laplace domain, which yields matrix equations of motion with constant coefficients. Some existing schemes are reviewed, the matrix Pade approximant is modified, and a technique which yields a minimal number of augmented states for a desired accuracy is presented. The state-space aeroelastic model is used to design an active control system for simultaneous flutter suppression and gust alleviation. The design target is for a continuous controller which transfers some measurements taken on the vehicle to a control command applied to a control surface. Structural modifications are formulated in a way which enables the treatment of passive flutter suppression system with the same procedures by which active control systems are designed.

  6. Maintenance of luminal pH and protease activity in lysosomes/late endosomes by vacuolar ATPase in chlorpromazine-treated RAW264 cells accumulating phospholipids.

    PubMed

    Hamaguchi, Ryohei; Haginaka, Jun; Tanimoto, Toshiko; Kuroda, Yukihiro

    2014-02-01

    Cationic amphiphilic drugs (CADs) inhibit phospholipases competitively/uncompetitively. It has also been reported that CADs spontaneously accumulate in acidic organelles and increase their luminal pH, which may lead to deactivation of phospholipid-metabolising enzymes, causing cellular phospholipid accumulation. Recently, however, contradictory results have also been reported in that the luminal pH is not increased by CAD treatment. In this study, we examined whether the lysosomal/late endosomal acidic pH was maintained by vacuolar ATPase (v-ATPase) after treatment with chlorpromazine (CPZ) as a model CAD. The activity of lysosomal protease after CPZ treatment was also measured. Oregon Green-dextran-tetramethylrhodamine conjugate was employed to determine the luminal pH of the lysosomes/late endosomes in RAW264 cells. The luminal pH remained acidic after treatment with CPZ for 23 h, and the lysosomal protease activity was not decreased by 5-min CPZ treatment. Co-treatment with CPZ and bafilomycin A1 (v-ATPase inhibitor) raised the luminal pH. These results suggest that the lysosomal/late endosomal pH is not affected by a 23-h CPZ treatment. In addition, lysosomal enzymes presumably maintain their activity when CPZ accumulates. Our results imply that the pH homeostasis in lysosomes/late endosomes is strictly maintained even after a longer treatment with CADs.

  7. Characterization of glutamate decarboxylase from Lactobacillus plantarum and its C-terminal function for the pH dependence of activity.

    PubMed

    Shin, Sun-Mi; Kim, Hana; Joo, Yunhye; Lee, Sang-Jae; Lee, Yong-Jik; Lee, Sang Jun; Lee, Dong-Woo

    2014-12-17

    The gadB gene encoding glutamate decarboxylase (GAD) from Lactobacillus plantarum was cloned and expressed in Escherichia coli. The recombinant enzyme exhibited maximal activity at 40 °C and pH 5.0. The 3D model structure of L. plantarum GAD proposed that its C-terminal region (Ile454-Thr468) may play an important role in the pH dependence of catalysis. Accordingly, C-terminally truncated (Δ3 and Δ11 residues) mutants were generated and their enzyme activities compared with that of the wild-type enzyme at different pH values. Unlike the wild-type GAD, the mutants showed pronounced catalytic activity in a broad pH range of 4.0-8.0, suggesting that the C-terminal region is involved in the pH dependence of GAD activity. Therefore, this study may provide effective target regions for engineering pH dependence of GAD activity, thereby meeting industrial demands for the production of γ-aminobutyrate in a broad range of pH values.

  8. Semimetallic MoP2: an active and stable hydrogen evolution electrocatalyst over the whole pH range

    NASA Astrophysics Data System (ADS)

    Pu, Zonghua; Saana Amiinu, Ibrahim; Wang, Min; Yang, Yushi; Mu, Shichun

    2016-04-01

    Developing efficient non-precious metal hydrogen evolution reaction (HER) electrocatalysts is a great challenge for sustainable hydrogen production from water. In this communication, for the first time, semimetallic MoP2 nanoparticle films on a metal Mo plate (MoP2 NPs/Mo) are fabricated through a facile two-step strategy. When used as a binder-free hydrogen evolution cathode, the as-prepared MoP2 NPs/Mo electrode exhibits superior HER catalytic activity at all pH values. At a current density of 10 mA cm-2, the catalyst displays overpotentials of 143, 211 and 194 mV in 0.5 M H2SO4, 1.0 M phosphate buffer solution and 1.0 M KOH, respectively. Furthermore, it exhibits excellent stability over a wide pH range. Thus, this in situ route opens up a new avenue for the fabrication of highly efficient, cost-effective and binder-free non-precious catalysts for water splitting and other electrochemical devices.Developing efficient non-precious metal hydrogen evolution reaction (HER) electrocatalysts is a great challenge for sustainable hydrogen production from water. In this communication, for the first time, semimetallic MoP2 nanoparticle films on a metal Mo plate (MoP2 NPs/Mo) are fabricated through a facile two-step strategy. When used as a binder-free hydrogen evolution cathode, the as-prepared MoP2 NPs/Mo electrode exhibits superior HER catalytic activity at all pH values. At a current density of 10 mA cm-2, the catalyst displays overpotentials of 143, 211 and 194 mV in 0.5 M H2SO4, 1.0 M phosphate buffer solution and 1.0 M KOH, respectively. Furthermore, it exhibits excellent stability over a wide pH range. Thus, this in situ route opens up a new avenue for the fabrication of highly efficient, cost-effective and binder-free non-precious catalysts for water splitting and other electrochemical devices. Electronic supplementary information (ESI) available: Experimental section and figures. See DOI: 10.1039/c6nr00820h

  9. Properties of acid whey as a function of pH and temperature.

    PubMed

    Chandrapala, Jayani; Duke, Mikel C; Gray, Stephen R; Zisu, Bogdan; Weeks, Mike; Palmer, Martin; Vasiljevic, Todor

    2015-07-01

    Compositional differences of acid whey (AW) in comparison with other whey types limit its processability and application of conventional membrane processing. Hence, the present study aimed to identify chemical and physical properties of AW solutions as a function of pH (3 to 10.5) at 4 different temperatures (15, 25, 40, or 90°C) to propose appropriate membrane-processing conditions for efficient use of AW streams. The concentration of minerals, mainly calcium and phosphate, and proteins in centrifuged supernatants was significantly lowered with increase in either pH or temperature. Lactic acid content decreased with pH decline and rose at higher temperatures. Calcium appeared to form complexes with phosphates and lactates mainly, which in turn may have induced molecular attractions with the proteins. An increase in pH led to more soluble protein aggregates with large particle sizes. Surface hydrophobicity of these particles increased significantly with temperature up to 40°C and decreased with further heating to 90°C. Surface charge was clearly pH dependent. High lactic acid concentrations appeared to hinder protein aggregation by hydrophobic interactions and may also indirectly influence protein denaturation. Processing conditions such as pH and temperature need to be optimized to manipulate composition, state, and surface characteristics of components of AW systems to achieve an efficient separation and concentration of lactic acid and lactose.

  10. [Effects of root-knot nematodes on cucumber leaf N and P contents, soil pH, and soil enzyme activities].

    PubMed

    Xu, Hua; Ruan, Wei-Bin; Gao, Yu-Bao; Song, Xiao-Yan; Wei, Yu-Kun

    2010-08-01

    A pot experiment was conducted to study the effects of inoculation with root-knot nematodes on the cucumber leaf N and P contents, and the rhizospheric and non-rhizospheric soil pH and enzyme activities. The rhizospheric soil pH didn't have a significant decrease until the inoculation rate reached 6000 eggs per plant. With the increase of inoculation rate, the leaf N and P contents, rhizospheric soil peroxidase activity, and rhizospheric and non-rhizospheric soil polyphenol oxidase activity all decreased gradually, rhizospheric soil catalase activity was in adverse, non-rhizospheric soil pH decreased after an initial increase, and non-rhizospheric soil catalase activity had no regular change. After inoculation, rhizospheric soil urease activity decreased significantly, but rhizospheric and non-rhizospheric soil phosphatase activity and non-rhizospheric soil peroxidase activity only had a significant decrease under high inoculation rate. In most cases, there existed significant correlations between rhizospheric soil pH, enzyme activities, and leaf N and P contents; and in some cases, there existed significant correlations between non-rhizospheric soil pH, enzyme activities, and leaf N and P contents.

  11. The respiratory burst activity and expression of catalase in white shrimp, Litopenaeus vannamei, during long-term exposure to pH stress.

    PubMed

    Wang, Wei-Na; Li, Bao-Sheng; Liu, Jin-Jian; Shi, Lei; Alam, M J; Su, Shi-Juan; Wu, Juan; Wang, Lei; Wang, An-Li

    2012-08-01

    In this study, changes of reactive oxygen species (ROS) and the mRNA expression of catalase of the Pacific white shrimp, Litopenaeus vannamei, exposed to pH (5.4, 6.7, 8.0, and 9.3) stress was investigated at different stress time (24, 48, 72, 96, and 120 h). Level of malondialdehyde (MDA) in shrimp also were assessed. The results revealed that acidic (pH 5.4 and 6.7) or alkaline exposure (pH 9.3) induced production of ROS hemocytes and increase of MDA level in shrimp. Moreover, the catalase mRNA expression in hepatopancreas of L. vannamei was up-regulated in 24 h at pH 5.4, in 72 h at pH 6.7 and in 48 h at pH 9.3, whereas was down-regulated significantly after 72 h acidic (pH 5.4 and 6.7) or alkaline (pH 9.4) exposure. In the present study, there was the relationship between ROS and catalase mRNA expression under normal acidic and alkaline conditions. At pH 8, the increase of catalase transcripts due to up-regulation by ROS, whereas MDA level did not significantly change, suggesting activation of corresponding protective mechanisms of detoxifying ROS is essential for the proper functioning of cells and the survival of shrimps.

  12. Characterization of steady-state activities of cytochrome c oxidase at alkaline pH: mimicking the effect of K-channel mutations in the bovine enzyme.

    PubMed

    Riegler, David; Shroyer, Lois; Pokalsky, Christine; Zaslavsky, Dmitry; Gennis, Robert; Prochaska, Lawrence J

    2005-01-07

    The cytochrome c oxidase activity of the bovine heart enzyme decreases substantially at alkaline pH, from 650 s(-1) at pH 7.0 to less than 10 s(-1) at pH 9.75. In contrast, the cytochrome c peroxidase activity of the enzyme shows little or no pH dependence (30-50 s(-1)) at pH values greater than 8.5. Under the conditions employed, it is demonstrated that the dramatic decrease in oxidase activity at pH 9.75 is fully reversible and not due to a major alkaline-induced conformational change in the enzyme. Furthermore, the Km values for cytochrome c interaction with the enzyme were also not significantly different at pH 7.8 and pH 9.75, suggesting that the pH dependence of the activity is not due to an altered interaction with cytochrome c at alkaline pH. However, at alkaline pH, the steady-state reduction level of the hemes increased, consistent with a slower rate of electron transfer from heme a to heme a3 at alkaline pH. Since it is well established that the rate of electron transfer from heme a to heme a3 is proton-coupled, it is reasonable to postulate that at alkaline pH, proton uptake becomes rate-limiting. The fact that this is not observed when hydrogen peroxide is used as a substrate in place of O2 suggests that the rate-limiting step is proton uptake via the K-channel associated with the reduction of the heme a3/CuB center prior to the reaction with O2. This step is not required for the reaction with H2O2, as shown previously in the examination of mutants of bacterial oxidases in which the K-channel was blocked. It is concluded that at pH values near 10, the delivery of protons via the K-channel becomes the rate-limiting step in the catalytic cycle with O2, so that the behavior of the bovine enzyme resembles that of the K-channel mutants in the bacterial enzymes.

  13. Accomplishments of the Abrupt Wing Stall (AWS) Program and Future Research Requirements

    NASA Technical Reports Server (NTRS)

    Hall, Robert M.; Woodson, Shawn H.; Chambers, Joseph R.

    2003-01-01

    The Abrupt Wing Stall (AWS) Program has addressed the problem of uncommanded lateral motions, such as wing drop and wing rock, at transonic speeds. The genesis of this Program was the experience of the F/A-18E/F Program in the late 199O's, when wing drop was discovered in the heart of the maneuver envelope for the pre-production aircraft. While the F/A-18E/F problem was subsequently corrected by a leading-edge flap scheduling change and the addition of a porous door to the wing fold fairing, the AWS Program was initiated as a national response to the lack of technology readiness available at the time of the F/A-18E/F Development Program. The AWS Program objectives were to define causal factors for the F/A-18E/F experience, to gain insights into the flow physics associated with wing drop, and to develop methods and analytical tools so that future programs could identify this type of problem before going to flight test. The paper reviews, for the major goals of the AWS Program, the status of the technology before the program began, the program objectives, accomplishments, and impacts. Lessons learned are presented for the benefit of future programs that must assess whether a vehicle will have uncommanded lateral motions before going to flight test. Finally, recommended future research needs are presented in light of the AWS Program experience.

  14. Alkali metals in addition to acidic pH activate the EvgS histidine kinase sensor in Escherichia coli.

    PubMed

    Eguchi, Yoko; Utsumi, Ryutaro

    2014-09-01

    Two-component signal transduction systems (TCSs) in bacteria perceive environmental stress and transmit the information via phosphorelay to adjust multiple cellular functions for adaptation. The EvgS/EvgA system is a TCS that confers acid resistance to Escherichia coli cells. Activation of the EvgS sensor initiates a cascade of transcription factors, EvgA, YdeO, and GadE, which induce the expression of a large group of acid resistance genes. We searched for signals activating EvgS and found that a high concentration of alkali metals (Na(+), K(+)) in addition to low pH was essential for the activation. EvgS is a histidine kinase, with a large periplasmic sensor region consisting of two tandem PBPb (bacterial periplasmic solute-binding protein) domains at its N terminus. The periplasmic sensor region of EvgS was necessary for EvgS activation, and Leu152, located within the first PBPb domain, was involved in the activation. Furthermore, chimeras of EvgS and PhoQ histidine kinases suggested that alkali metals were perceived at the periplasmic sensor region, whereas the cytoplasmic linker domain, connecting the transmembrane region and the histidine kinase domain, was required for low-pH perception.

  15. Effects of long-term pH elevation on the sulfate-reducing and methanogenic activities of anaerobic sewer biofilms.

    PubMed

    Gutierrez, Oriol; Park, Donghee; Sharma, Keshab Raj; Yuan, Zhiguo

    2009-05-01

    The dosage of alkali is often applied by the wastewater industry to reduce the transfer of hydrogen sulfide from wastewater to the sewer atmosphere. In this paper the activities of Sulfate Reducing Bacteria (SRB) and Methanogenic Archaea (MA) under elevated pH conditions (8.6 and 9.0) were evaluated in a laboratory scale anaerobic sewer reactor. Compared to those in a control reactor without pH control (pH 7.6+/-0.1), the SRB activity was reduced by 30% and 50%, respectively, at pH 8.6 and pH 9.0. When normal pH was resumed, it took approximately 1 month for the SRB activity to fully recover. Methanogenic activities developed in the control reactor in 3 months after the reactor start-up, while no significant methanogenic activities were detected in the experimental reactor until normal pH was resumed. The results suggest that elevated pH at 8.6-9.0 suppressed the growth of methanogens. These experimental results clearly showed that, in addition to its well-known effect of reducing H(2)S transfer from the liquid to the gas phase, pH elevation considerably reduces sulfide and methane production by anaerobic sewer biofilms. These findings are significant for the optimal use of alkali addition to sewers for the control of H(2)S and CH(4) emissions. A model-based study showed that, by adding the alkali at the beginning rather than towards the end of a rising main, substantial savings in chemicals can be achieved while achieving the same level of sulfide emission control, and complete methane emission control.

  16. Carbonic Anhydrase Activity Monitored In Vivo by Hyperpolarized 13C-Magnetic Resonance Spectroscopy Demonstrates Its Importance for pH Regulation in Tumors.

    PubMed

    Gallagher, Ferdia A; Sladen, Helen; Kettunen, Mikko I; Serrao, Eva M; Rodrigues, Tiago B; Wright, Alan; Gill, Andrew B; McGuire, Sarah; Booth, Thomas C; Boren, Joan; McIntyre, Alan; Miller, Jodi L; Lee, Shen-Han; Honess, Davina; Day, Sam E; Hu, De-En; Howat, William J; Harris, Adrian L; Brindle, Kevin M

    2015-10-01

    Carbonic anhydrase buffers tissue pH by catalyzing the rapid interconversion of carbon dioxide (CO2) and bicarbonate (HCO3 (-)). We assessed the functional activity of CAIX in two colorectal tumor models, expressing different levels of the enzyme, by measuring the rate of exchange of hyperpolarized (13)C label between bicarbonate (H(13)CO3(-)) and carbon dioxide ((13)CO2), following injection of hyperpolarized H(13)CO3(-), using (13)C-magnetic resonance spectroscopy ((13)C-MRS) magnetization transfer measurements. (31)P-MRS measurements of the chemical shift of the pH probe, 3-aminopropylphosphonate, and (13)C-MRS measurements of the H(13)CO3(-)/(13)CO2 peak intensity ratio showed that CAIX overexpression lowered extracellular pH in these tumors. However, the (13)C measurements overestimated pH due to incomplete equilibration of the hyperpolarized (13)C label between the H(13)CO3(-) and (13)CO2 pools. Paradoxically, tumors overexpressing CAIX showed lower enzyme activity using magnetization transfer measurements, which can be explained by the more acidic extracellular pH in these tumors and the decreased activity of the enzyme at low pH. This explanation was confirmed by administration of bicarbonate in the drinking water, which elevated tumor extracellular pH and restored enzyme activity to control levels. These results suggest that CAIX expression is increased in hypoxia to compensate for the decrease in its activity produced by a low extracellular pH and supports the hypothesis that a major function of CAIX is to lower the extracellular pH.

  17. Carbonic anhydrase activity monitored in vivo by hyperpolarized 13C-magnetic resonance spectroscopy demonstrate its importance for pH regulation in tumors

    PubMed Central

    Gallagher, Ferdia A.; Sladen, Helen; Kettunen, Mikko I.; Serrao, Eva M.; Rodrigues, Tiago B.; Wright, Alan; Gill, Andrew B.; McGuire, Sarah; Booth, Thomas C.; Boren, Joan; McIntyre, Alan; Miller, Jodi L.; Lee, Shen-Han; Honess, Davina; Day, Sam E.; Hu, De-en; Howat, William J.; Harris, Adrian L.; Brindle, Kevin M.

    2015-01-01

    Carbonic anhydrase (CA) buffers tissue pH by catalyzing the rapid interconversion of carbon dioxide (CO2) and bicarbonate (HCO3−). We assessed the functional activity of CAIX in two colorectal tumor models, expressing different levels of the enzyme, by measuring the rate of exchange of hyperpolarized 13C label between bicarbonate (H13CO3−) and carbon dioxide (13CO2), following injection of hyperpolarized H13CO3−, using 13C magnetic resonance spectroscopy (13C-MRS) magnetization transfer measurements. 31P-MRS measurements of the chemical shift of the pH probe, 3-aminopropylphosphonate, and 13C-MRS measurements of the H13CO3−/13CO2 peak intensity ratio showed that CAIX overexpression lowered extracellular pH in these tumors. However, the 13C measurements overestimated pH due to incomplete equilibration of the hyperpolarized 13C label between the H13CO3− and 13CO2 pools. Paradoxically, tumors overexpressing CAIX showed lower enzyme activity using magnetization transfer measurements, which can be explained by the more acidic extracellular pH in these tumors and the decreased activity of the enzyme at low pH. This explanation was confirmed by administration of bicarbonate in the drinking water, which elevates tumor extracellular pH and restored enzyme activity to control levels. These results suggest that CAIX expression is increased in hypoxia to compensate for the decrease in its activity produced by a low extracellular pH, and supports the hypothesis that a major function of CAIX is to lower the extracellular pH. PMID:26249175

  18. [Effect of pH of Adsorption Buffers on the Number and Antigen-Binding Activity of Monoclonal Antibodies Immobilized on the Surface of Polystyrene Microplates].

    PubMed

    Tarakanova, Yu N; Dmitriev, A D; Massino, Yu S; Pechelulko, A A; Segal, O L; Skoblov, Yu O; Ulanova, T I; Lavrov, V F; Dmitriev, D A

    2015-01-01

    The change in the concentration and antigen-binding activity of 28 monoclonal antibodies was studied after their adsorption on the surface of polystyrene microplates in buffers with different pH values (1.0, 2.8, 7.5, 9.6, and 11.9). We used 16 clones to the HIV p24 protein and 12 clones to the surface antigen of Hepatitis B Virus. The binding efficiency of adsorbed antibodies to the labeled antigen was evaluated by the slope of the linear region of the binding curve to the concentration axis. It was shown that the antigen-binding activity of six antibodies (21.5%) statistically significantly increased after adsorption at pH 2.8 and 11.9 as compared to pH 7.5 and 9.5. The maximum amount of antibodies was found to be adsorbed on the solid surface at pH 7.5. The analysis of the binding of 125I-HBs-antigen to adsorbed antibodies made it possible to evaluate the concentration of active antibodies on the polystyrene surface. It was shown that the increase in the antigen-binding activity was due to an increase in the proportion of antibodies with retained activity after adsorption at pH 2.8 and 11.9. Under these conditions, about 20% of the antibodies retained their antigen-binding activity, and 6% did so after immobilization at pH 7.5.

  19. Intracellular pH regulation in unstimulated Calliphora salivary glands is Na+ dependent and requires V-ATPase activity.

    PubMed

    Schewe, Bettina; Blenau, Wolfgang; Walz, Bernd

    2012-04-15

    Salivary gland cells of the blowfly Calliphora vicina have a vacuolar-type H(+)-ATPase (V-ATPase) that lies in their apical membrane and energizes the secretion of a KCl-rich primary saliva upon stimulation with serotonin (5-hydroxytryptamine). Whether and to what extent V-ATPase contributes to intracellular pH (pH(i)) regulation in unstimulated gland cells is unknown. We used the fluorescent dye BCECF to study intracellular pH(i) regulation microfluorometrically and show that: (1) under resting conditions, the application of Na(+)-free physiological saline induces an intracellular alkalinization attributable to the inhibition of the activity of a Na(+)-dependent glutamate transporter; (2) the maintenance of resting pH(i) is Na(+), Cl(-), concanamycin A and DIDS sensitive; (3) recovery from an intracellular acid load is Na(+) sensitive and requires V-ATPase activity; (4) the Na(+)/H(+) antiporter is not involved in pH(i) recovery after a NH(4)Cl prepulse; and (5) at least one Na(+)-dependent transporter and the V-ATPase maintain recovery from an intracellular acid load. Thus, under resting conditions, the V-ATPase and at least one Na(+)-dependent transporter maintain normal pH(i) values of pH 7.5. We have also detected the presence of a Na(+)-dependent glutamate transporter, which seems to act as an acid loader. Despite this not being a common pH(i)-regulating transporter, its activity affects steady-state pH(i) in C. vicina salivary gland cells.

  20. Evaluation of thiol Raman activities and pKa values using internally referenced Ramanbased pH titration

    NASA Astrophysics Data System (ADS)

    Suwandaratne, Nuwanthi

    Thiols are one of the most important classes of chemicals used broadly in organic synthesis, biological chemistry, and nanosciences. Thiol pKa values are key indicators of thiol reactivity and functionality. This study is an internally-referenced Raman-based pH titration method that enables reliable quantification of thiol pKa values for both mono- and di-thiols in water. The degree of thiol ionization is monitored directly using the peak intensity of the S-H stretching feature relative to an internal reference peak as a function of solution pH. The thiol pKa values and Raman activity relative to its internal reference were then determined by curve-fitting the experimental data with equations derived on the basis of the Henderson-Hasselbalch equation. Using this Raman titration method, first and second thiol pKa values for 1,2-benzenedithol in water were determined for the first time. This method is convenient to implement and its underlying theory is easy to follow.

  1. Semimetallic MoP2: an active and stable hydrogen evolution electrocatalyst over the whole pH range.

    PubMed

    Pu, Zonghua; Saana Amiinu, Ibrahim; Wang, Min; Yang, Yushi; Mu, Shichun

    2016-04-28

    Developing efficient non-precious metal hydrogen evolution reaction (HER) electrocatalysts is a great challenge for sustainable hydrogen production from water. In this communication, for the first time, semimetallic MoP2 nanoparticle films on a metal Mo plate (MoP2 NPs/Mo) are fabricated through a facile two-step strategy. When used as a binder-free hydrogen evolution cathode, the as-prepared MoP2 NPs/Mo electrode exhibits superior HER catalytic activity at all pH values. At a current density of 10 mA cm(-2), the catalyst displays overpotentials of 143, 211 and 194 mV in 0.5 M H2SO4, 1.0 M phosphate buffer solution and 1.0 M KOH, respectively. Furthermore, it exhibits excellent stability over a wide pH range. Thus, this in situ route opens up a new avenue for the fabrication of highly efficient, cost-effective and binder-free non-precious catalysts for water splitting and other electrochemical devices.

  2. Surface Properties and Photocatalytic Activities of the Colloidal ZnS:Mn Nanocrystals Prepared at Various pH Conditions

    PubMed Central

    Heo, Jungho; Hwang, Cheong-Soo

    2015-01-01

    Water-dispersible ZnS:Mn nanocrystals (NC) were synthesized by capping the surface with mercaptoacetic acid (MAA) molecules at three different pH conditions. The obtained ZnS:Mn-MAA NC products were physically and optically characterized by corresponding spectroscopic methods. The UV-Visible absorption spectra and PL emission spectra showed broad peaks at 310 and 590 nm, respectively. The average particle sizes measured from the HR-TEM images were 5 nm, which were also supported by the Debye-Scherrer calculations using the X-ray diffraction (XRD) data. Moreover, the surface charges and the degrees of aggregation of the ZnS:Mn-MAA NCs were determined by electrophoretic and hydrodynamic light scattering methods, indicating formation of agglomerates in water with various sizes (50–440 nm) and different surface charge values accordingly the preparation conditions of the NCs (−7.59 to −24.98 mV). Finally, the relative photocatalytic activities of the ZnS:Mn-MAA NCs were evaluated by measuring the degradation rate of methylene blue (MB) molecule in a pseudo first-order reaction condition under the UV-visible light irradiation. As a result, the ZnS:Mn-MAA NC prepared at the pH 7 showed the best photo-degradation efficiency of the MB molecule with the first-order rate constant (kobs) of 2.0 × 10−3·min−1. PMID:28347105

  3. Role of pH in the formation of structurally stable and catalytically active supported gold catalysts

    SciTech Connect

    Veith, Gabriel M; Lupini, Andrew R; Dudney, Nancy J

    2009-01-01

    We report the investigation of titania (Degussa P25) supported gold catalysts prepared by magnetron sputtering. Catalysts grown on natural fumed titania were structurally unstable, resulting in the rapid coarsening of 2.4 nm gold clusters into large {approx}20 nm gold clusters in a few days at room temperature under normal atmospheric conditions. However, treating the titania support powder to a mock deposition-precipitation process, at pH 4, followed by the subsequent deposition of gold onto this treated powder produced a remarkable enhancement in gold particle stability and a 20-fold enhancement of catalytic activity. Furthermore, it was found that treating the titania under basic conditions (pH 10) resulted in a further enhancement of structural stability and a further doubling of the reaction rate to 0.28 mol of CO/mol of Au {center_dot} s. This enhancement cannot be attributed to removing surface Cl{sup -} species from the titania, the formation of oxygen vacancies on the TiO{sub 2} surface, or an electronic effect. Instead, it appears to be associated with the formation of strongly bound hydroxyl species on the TiO{sub 2} surface. The formation of surface hydroxyls during the deposition-precipitation method is coincidental and contributes significantly to the properties of Au/TiO{sub 2} catalysts.

  4. Effect of thymoquinone on cytosolic pH and Na+/H+ exchanger activity in mouse dendritic cells.

    PubMed

    Yang, Wenting; Bhandaru, Madhuri; Pasham, Venkanna; Bobbala, Diwakar; Zelenak, Christine; Jilani, Kashif; Rotte, Anand; Lang, Florian

    2012-01-01

    The anti-inflammatory Nigella sativa component thymoquinone compromises the function of dendritic cells (DCs), key players in the regulation of innate and adaptive immunity. DC function is regulated by the Na(+)/H(+) exchanger (NHE), which is stimulated by lipopolysaccharides (LPS) and required for LPS-induced cell swelling, reactive oxygen species (ROS) production, TNF-α release and migration. Here we explored, whether thymoquinone influences NHE activity in DCs. To this end, bone marrow derived mouse DCs were treated with LPS in the absence and presence of thymoquinone (10 μM). Cytosolic pH (pH(i)) was determined from 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) fluorescence, NHE activity from the Na(+)-dependent realkalinization following an ammonium pulse, cell volume from forward scatter in FACS analysis, ROS production from 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) fluorescence, TNF-α production utilizing ELISA and DC migration with transwell migration assays. As a result, exposure of DCs to LPS (1 μg/ml) led within 4 hours to transient increase of NHE activity. Thymoquinone did not significantly modify cytosolic pH or cellular NHE activity in the absence of LPS, but abrogated the effect of LPS on NHE activity. Accordingly, in the presence of thymoquinone LPS-treatment resulted in cytosolic acidification. LPS further increased forward scatter and ROS formation, effects similarly abrogated by thymoquinone. Again, in the absence of LPS, thymoquinone did not significantly modify ROS formation and cell volume. LPS further triggered TNF-α release and migration, effects again blunted in the presence of thymoquinone. NHE1 inhibitor cariporide (10 μM) blunted LPS induced TNF-α release and migration. The effects of thymoquinone on NHE activity and migration were reversed upon treatment of the cells with t-butyl hydroperoxide (TBOOH, 5 μM). In conclusion, thymoquinone blunts LPS induced NHE activity, cell swelling, oxidative burst

  5. NIR technology for on-line determination of superficial a(w) and moisture content during the drying process of fermented sausages.

    PubMed

    Collell, Carles; Gou, Pere; Arnau, Jacint; Muñoz, Israel; Comaposada, Josep

    2012-12-01

    Three different NIR equipment were evaluated based on their ability to predict superficial water activity (a(w)) and moisture content in two types of fermented sausages (with and without moulds on surface), using partial least squares (PLS) regression models. The instruments differed mainly in wavelength range, resolution and measurement configuration. The most accurate equipment was used in a new experiment to achieve robust models in sausages with different salt contents and submitted to different drying conditions. The models developed showed determination coefficients (R(2)(P)) values of 0.990, 0.910 and 0.984, and RMSEP values of 1.560%, 0.220% and 0.007% for moisture, salt and a(w) respectively. It was demonstrated that NIR spectroscopy could be a suitable non-destructive method for on-line monitoring and control of the drying process in fermented sausages.

  6. Effect of pH and HNO2 concentration on the activity of ammonia-oxidizing bacteria in a partial nitritation reactor.

    PubMed

    Claros, J; Jiménez, E; Aguado, D; Ferrer, J; Seco, A; Serralta, J

    2013-01-01

    Ammonia-oxidizing bacteria (AOB) are very sensitive to environmental conditions and wastewater treatment plant operational parameters. One of the most important factors affecting their activity is pH. Its effect is associated with: NH3/NH4(+) and HNO2/NO2(-) chemical equilibriums and biological reaction rates. The aim of this study was to quantify and model the effect of pH and free nitrous acid (FNA) concentration on the activity of AOB present in a lab-scale partial nitritation reactor. For this purpose, two sets of batch experiments were carried out using biomass from this reactor. Fluorescent in situ hybridization analysis showed that Nitrosomona eutropha and Nitrosomona europaea species were dominant in the partial nitritation reactor (>94%). The experimental results showed that FNA inhibits the AOB activity. This inhibition was properly modelled by the non-competitive inhibition function and the half inhibition constant value was determined as 1.32 mg HNO2-N L(-1). The optimal pH for these AOB was found to be in the range 7.4-7.8. The pH inhibitory effect was stronger at high pH values than at low pH values. Therefore, an asymmetric inhibition function was proposed to represent the pH effect on these bacteria. A combination of two sigmoidal functions was able to reproduce the experimental results obtained.

  7. Activated RhoA Binds to the Pleckstrin Homology (PH) Domain of PDZ-RhoGEF, a Potential Site for Autoregulation

    SciTech Connect

    Chen, Zhe; Medina, Frank; Liu, Mu-ya; Thomas, Celestine; Sprang, Stephen R.; Sternweis, Paul C.

    2010-07-19

    Guanine nucleotide exchange factors (GEFs) catalyze exchange of GDP for GTP by stabilizing the nucleotide-free state of the small GTPases through their Dbl homology/pleckstrin homology (DH {center_dot} PH) domains. Unconventionally, PDZ-RhoGEF (PRG), a member of the RGS-RhoGEFs, binds tightly to both nucleotide-free and activated RhoA (RhoA {center_dot} GTP). We have characterized the interaction between PRG and activated RhoA and determined the structure of the PRG-DH {center_dot} PH-RhoA {center_dot} GTP{gamma}S (guanosine 5{prime}-O-[{gamma}-thio]triphosphate) complex. The interface bears striking similarity to a GTPase-effector interface and involves the switch regions in RhoA and a hydrophobic patch in PRG-PH that is conserved among all Lbc RhoGEFs. The two surfaces that bind activated and nucleotide-free RhoA on PRG-DH {center_dot} PH do not overlap, and a ternary complex of PRG-DH {center_dot} PH bound to both forms of RhoA can be isolated by size-exclusion chromatography. This novel interaction between activated RhoA and PH could play a key role in regulation of RhoGEF activity in vivo.

  8. Effects of heat and pH treatments and in vitro digestion on the biological activity of protein hydrolysates of Amaranthus hypochondriacus L. grain.

    PubMed

    López-Sánchez, J; Ponce-Alquicira, E; Pedroza-Islas, R; de la Peña-Díaz, A; Soriano-Santos, J

    2016-12-01

    The aim of this work was to assess the effects of temperature (T), time (t) and pH treatments and an in vitro digestion on the stability of the angiotensin I-converting-enzyme-inhibitory activity (ACEIA) and antithrombotic activity (ATA; assessed as inhibition of platelet aggregation) of selected protein hydrolysates of amaranth named Alb1H103 and GloH88 and GluH24 with dipeptidyl peptidase IV inhibitory activity (DPPIVIA). Heat treatment (40-100 °C) for 1 h showed no significant differences among ACEIA, DPPIVIA and ATA of the heated hydrolysates at pH 4 and 7. There was no statistically significant loss of any bioactivity under heat treatment for 3 h at pH 4.0. Alb1H103 and GluH24 maintained the inhibitory activity of ACE and ATA at pH 7.0 for 3 h, whereas GloH88 maintained ACEIA and ATA for 2.0 h at pH 7.0. The pH effect on hydrolysates bioactivity was assessed in the range of 2.0-12.0. This was negligible on ACEIA, ATA and DPPIVIA. The in vitro digestion was performed using pepsin, trypsin (T) and α-chymotrypsin (C). A previous treatment of hydrolysates with pepsin improved the proteolytic activities of T and C. The hydrolysates kept at 100 °C for 1 h at pH 4.0, showed a significant increase in bioactivity. Conversely, a treatment at pH 7.0 showed no significant difference (p < 0.05) in the hydrolysates bioactivities after their digestion. Thus, biological activity of hydrolysates may be preserved or enhanced, depending on their processing conditions.

  9. ATP-mediated intrinsic peroxidase-like activity of Fe3O4-based nanozyme: One step detection of blood glucose at physiological pH.

    PubMed

    Vallabani, N V Srikanth; Karakoti, Ajay S; Singh, Sanjay

    2017-05-01

    Fe3O4 nanoparticles (Fe3O4 NPs), demonstrating peroxidase-like activity has garnered attention in the detection of several biomolecules, therefore, emerged as an excellent nano-biosensing agent. The intrinsic peroxidase-like activity of Fe3O4 NPs at acidic pH is the fundamental action driving the oxidation of substrates like TMB, resulting in a colorimetric product formation used in the detection of biomolecules. Hence, the detection sensitivity essentially depends on the ability of oxidation by Fe3O4 NPs in presence of H2O2. However, the limited sensitivity and pH condition constraint have been identified as the major drawbacks in the detection of biomolecules at physiological pH. Herein, we report overwhelming of the fundamental limitation of acidic pH and tuning the peroxidase-like activity of Fe3O4 NPs at physiological pH by using ATP. In presence of ATP, Fe3O4 NPs exhibited enhanced peroxidase-like activity over a wide range of pH and temperatures. Mechanistically, it was found that the ability of ATP to participate in single electron transfer reaction, through complexation with Fe3O4 NPs, results in the generation of hydroxyl radicals which are responsible for enhanced peroxidase activity at physiological pH. We utilized this ATP-mediated enhanced peroxidase-like activity of Fe3O4 NPs for single step detection of glucose with a colorimetric detection limit of 50μM. Further, we extended this single step detection method to monitor glucose level in human blood serum and detected in a time span of <5min at pH 7.4.

  10. Acoustic wave (AW) based moisture sensor for use with corrosive gases

    DOEpatents

    Pfeifer, K.B.; Frye, G.C.; Schneider, T.W.

    1996-11-05

    Moisture corrosive gas stream is measured as a function of the difference in resonant frequencies between two acoustic wave (AW) devices, each with a film which accepts at least one of the components of the gas stream. One AW is located in the gas stream while the other is located outside the gas stream but in the same thermal environment. In one embodiment, the film is a hydrophilic material such as SiO{sub 2}. In another embodiment, the SiO{sub 2} is covered with another film which is impermeable to the corrosive gas, such that the AW device in the gas stream measures only the water vapor. In yet another embodiment, the film comprises polyethylene oxide which is hydrophobic and measures only the partial pressure of the corrosive gas. Other embodiments allow for compensation of drift in the system. 8 figs.

  11. Acoustic wave (AW) based moisture sensor for use with corrosive gases

    DOEpatents

    Pfeifer, Kent B.; Frye, Gregory C.; Schneider, Thomas W.

    1996-01-01

    Moisture corrosive gas stream is measured as a function of the difference in resonant frequencies between two acoustic wave (AW) devices, each with a film which accepts at least one of the components of the gas stream. One AW is located in the gas stream while the other is located outside the gas stream but in the same thermal environment. In one embodiment, the film is a hydrophilic material such as SiO.sub.2. In another embodiment, the SiO.sub.2 is covered with another film which is impermeable to the corrosive gas, such that the AW device in the gas stream measures only the water vapor. In yet another embodiment, the film comprises polyethylene oxide which is hydrophobic and measures only the partial pressure of the corrosive gas. Other embodiments allow for compensation of drift in the system.

  12. Intracellular carbonic anhydrase activity sensitizes cancer cell pH signaling to dynamic changes in CO2 partial pressure.

    PubMed

    Hulikova, Alzbeta; Aveyard, Nicholas; Harris, Adrian L; Vaughan-Jones, Richard D; Swietach, Pawel

    2014-09-12

    Carbonic anhydrase (CA) enzymes catalyze the chemical equilibration among CO2, HCO3(-) and H(+). Intracellular CA (CAi) isoforms are present in certain types of cancer, and growing evidence suggests that low levels correlate with disease severity. However, their physiological role remains unclear. Cancer cell CAi activity, measured as cytoplasmic CO2 hydration rate (kf), ranged from high in colorectal HCT116 (∼2 s(-1)), bladder RT112 and colorectal HT29, moderate in fibrosarcoma HT1080 to negligible (i.e. spontaneous kf = 0.18 s(-1)) in cervical HeLa and breast MDA-MB-468 cells. CAi activity in cells correlated with CAII immunoreactivity and enzymatic activity in membrane-free lysates, suggesting that soluble CAII is an important intracellular isoform. CAi catalysis was not obligatory for supporting acid extrusion by H(+) efflux or HCO3(-) influx, nor for maintaining intracellular pH (pHi) uniformity. However, in the absence of CAi activity, acid loading from a highly alkaline pHi was rate-limited by HCO3(-) supply from spontaneous CO2 hydration. In solid tumors, time-dependence of blood flow can result in fluctuations of CO2 partial pressure (pCO2) that disturb cytoplasmic CO2-HCO3(-)-H(+) equilibrium. In cancer cells with high CAi activity, extracellular pCO2 fluctuations evoked faster and larger pHi oscillations. Functionally, these resulted in larger pH-dependent intracellular [Ca(2+)] oscillations and stronger inhibition of the mTORC1 pathway reported by S6 kinase phosphorylation. In contrast, the pHi of cells with low CAi activity was less responsive to pCO2 fluctuations. Such low pass filtering would "buffer" cancer cell pHi from non-steady-state extracellular pCO2. Thus, CAi activity determines the coupling between pCO2 (a function of tumor perfusion) and pHi (a potent modulator of cancer cell physiology).

  13. Influence of temperature, pH and water activity on "in vitro" inhibition of Penicillium glabrum (Wehmer) Westling by yeasts.

    PubMed

    Sinigaglia, M; Corbo, M R; Ciccarone, C

    1998-08-01

    Four different yeast species (Metschnikowia pulcherrima, Saccharomycopsis vini, Kluyveromyces marxianus, Cryptococcus albidus), isolated from surface of grapes, were evaluated for biocontrol potential against Penicillium glabrum. In order to investigate the influence of temperature, pH, water activity and yeast cell concentration on Penicillium glabrum inhibition, the individual effects and the interaction of these factors were analyzed by means of a Central Composite Design (CCD). All yeast species tested showed antagonistic effects which were more pronounced at high cell concentrations. The other variables affected the antagonistic effect differentially depending on the yeast species. Results of the experimental design showed that the selective success of a competitive microflora is under environmental control; moreover, when microbial cells are subjected to multiple factors, the effects and the reciprocal interactions of the individual variables cannot be independently evaluated.

  14. A low-temperature-active alkaline pectate lyase from Xanthomonas campestris ACCC 10048 with high activity over a wide pH range.

    PubMed

    Yuan, Peng; Meng, Kun; Wang, Yaru; Luo, Huiying; Shi, Pengjun; Huang, Huoqing; Tu, Tao; Yang, Peilong; Yao, Bin

    2012-11-01

    Alkaline pectate lyases are favorable for the textile industry. Here, we report the gene cloning and expression of a low-temperature-active alkaline pectate lyase (PL D) from Xanthomonas campestris ACCC 10048. Deduced PL D consists of a putative 27-residue signal peptide and a catalytic domain of 320 residues belonging to family PF09492. Recombinant PL D (r-PL D) produced in Escherichia coli was purified to electrophoretic homogeneity with a single step of Ni(2+)-NTA affinity chromatography and showed an apparent molecular weight of ~38 kDa. The pH and temperature optima of r-PL D were found to be 9.0 °C and 30 °C, respectively. Compared with its microbial counterparts, r-PL D had higher activity over a wide pH range (>45 % of the maximum activity at pH 3.0-12.0) and at lower temperatures (>35 % of activity even at 0 °C). The K(m) and V(max) values of r-PL D for polygalacturonic acid were 4.9 gl(-1) and 30.1 μmolmin(-1) mg(-1), respectively. Compared with the commercial compound pectinase from Novozymes, r-PL D showed similar efficacy in reducing the intrinsic viscosity of polygalacturonic acid (35.1 % vs. 36.5 %) and in bioscouring of jute (10.25 % vs. 10.82 %). Thus, r-PL D is a valuable additive candidate for the textile industry.

  15. Outdoor cultures of Chlorella pyrenoidosa in the effluent of anaerobically digested activated sludge: The effects of pH and free ammonia.

    PubMed

    Tan, Xiao-Bo; Zhang, Ya-Lei; Yang, Li-Bin; Chu, Hua-Qiang; Guo, Jun

    2016-01-01

    A freshwater algae Chlorella pyrenoidosa was cultured outdoors using anaerobically digested activated sludge effluent. The effects of pH variations were evaluated. The coupled pH variations and free ammonia toxicity significantly affected the algal growth, lipids accumulation and contamination control during every season. The free ammonia toxicity at high pH levels actually inhibited the algal growth. Compared to an optimal algal growth at a pH of 5.7-6.5, biomass productivity at a high pH of 8.3-8.8 was reduced by 67.15±6.98%, 54.39±6.42% and 83.63±5.71% in the spring, fall and summer, respectively. When the pH rose above 9.1-9.6, algae were unable to grow in the wastewater. However, high pH levels reduced contamination (e.g., bacteria and microalgae grazers) and triggered lipids accumulation in algal cells. These findings suggest that pH control strategies are essential for this type of algal wastewater system, where ammonia is the dominant nitrogen source.

  16. Operational test report for 241-AW tank inlet air control stations

    SciTech Connect

    Minteer, D.J., Westinghouse Hanford

    1996-07-03

    This document reports the results of operational testing on tank inlet air control stations in 241-AW tank farm. An air control station was installed on each of the six AW tanks. Operational testing consisted of a simple functional test of each station`s air flow controller, aerosol testing of each station`s HEPA filter, and final ventilation system balancing (i.e., tank airflows and vacuum level) using the air control stations. The test was successful and the units were subsequently placed into operation.

  17. Peroxidase-like activity of gold nanoparticles stabilized by hyperbranched polyglycidol derivatives over a wide pH range

    NASA Astrophysics Data System (ADS)

    Drozd, Marcin; Pietrzak, Mariusz; Parzuchowski, Paweł; Mazurkiewicz-Pawlicka, Marta; Malinowska, Elżbieta

    2015-12-01

    The aim of this work was to carry out comparative studies on the peroxidase-like activity of gold nanoparticles (AuNPs) stabilized with low molecular weight hyperbranched polyglycidol (HBPG-OH) and its derivative modified with maleic acid residues (HBPG-COOH). The influence of the stabilizer to gold precursor ratio on the size and morphology of nanoparticles obtained was checked, and prepared nanoparticles were characterized by means of transmission electron microscopy and UV-Vis spectroscopy. The results indicated the divergent effect of increasing the concentration of stabilizers (HBPG-OH or HBPG-COOH) on the size of the nanostructures obtained. The gold nanoparticles obtained were characterized as having intrinsic peroxidase-like activity and the mechanism of catalysis in acidic and alkaline mediums was consistent with the standard Michaelis-Menten kinetics, revealing a strong affinity of AuNPs with 2, 2‧-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 3, 3‧, 5, 5‧-tetramethylbenzidine (TMB), and significantly lower affinity towards phenol. By comparing the kinetic parameters, a negligible effect of polymeric ligand charge on activity against various types of substrates (anionic or cationic) was indicated. The superiority of steric stabilization via the application of tested low-weight hyperbranched polymers over typical stabilizers in preventing salt-induced aggregation and maintaining high catalytic activity in time was proved. The applied hyperbranched stabilizers provide a good tool for manufacturing gold-based nanozymes, which are highly stable and active over a wide pH range.

  18. Production of two highly active bacterial phytases with broad pH optima in germinated transgenic rice seeds.

    PubMed

    Hong, Chwan-Yang; Cheng, Kuo-Joan; Tseng, Tung-Hai; Wang, Chang-Sheng; Liu, Li-Fei; Yu, Su-May

    2004-02-01

    Phytate is the main storage form of phosphorus in many plant seeds, but phosphate bound in this form is not available to monogastric animals. Phytase, an enzyme that hydrolyzes phosphate from phytate, has the potential to enhance phosphorus availability in animal diets when engineered in rice seeds as a feed additive. Two genes, derived from a ruminal bacterium Selenomonas ruminantium (SrPf6) and Escherichia coli (appA), encoding highly active phytases were expressed in germinated transgenic rice seeds. Phytase expression was controlled by a germination inducible alpha-amylase gene (alphaAmy8) promoter, and extracellular phytase secretion directed by an betaAmy8 signal peptide sequence. The two phytases were expressed in germinated transgenic rice seeds transiently and in a temporally controlled and tissue-specific manner. No adverse effect on plant development or seed formation was observed. Up to 0.6 and 1.4 U of phytase activity per mg of total extracted cellular proteins were obtained in germinated transgenic rice seeds expressing appA and SrPf6 phytases, respectively, which represent 46-60 times of phytase activities compared to the non-transformant. The appA and SrPf6 phytases produced in germinated transgenic rice seeds had high activity over broad pH ranges of 3.0-5.5 and 2.0-6.0, respectively. Phytase levels and inheritance of transgenes in one highly expressing plant were stable over four generations. Germinated transgenic rice seeds, which produce a highly active recombinant phytase and are rich in hydrolytic enzymes, nutrients and minerals, could potentially be an ideal feed additive for improving the phytate-phosphorus digestibility in monogastric animals.

  19. Effect of dietary roughage level on chewing activity, ruminal pH, and saliva secretion in lactating Holstein cows.

    PubMed

    Jiang, F G; Lin, X Y; Yan, Z G; Hu, Z Y; Liu, G M; Sun, Y D; Liu, X W; Wang, Z H

    2017-04-01

    Increasing dietary roughage level is a commonly used strategy to prevent subacute ruminal acidosis. We hypothesized that high-roughage diets could promote chewing activity, saliva secretion, and hence more alkaline to buffer rumen pH. To verify the hypothesis, 12 multiparous Holstein cows in mid lactation were randomly allocated to 4 treatments in a triplicated 4 × 4 Latin square experiment with one cow in each treatment surgically fitted with a ruminal cannula. Treatments were diets containing 40, 50, 60, or 70% of roughage on a DM basis. Increasing dietary roughage level decreased DM, CP, OM, starch, and NEL intake, increased ADF intake, and decreased milk yield linearly. Intake of NDF was quite stable across treatments and ranged from 7.8 to 8.1 kg/d per cow. Daily eating time increased linearly with increased roughage level. The increase in eating time was due to increased eating time per meal but not number of meals per day, which was stable and ranged from 8.3 to 8.5 meals per day across treatments. Increasing dietary roughage level had no effect on ruminating time (min/d), the number of ruminating periods (rumination periods per d), and chewing time per ruminating period (min/ruminating period). Ruminating time per kilogram of NDF intake and total chewing time per kilogram of ADF intake were similar across treatments (57.4 and 183.8 min/kg, respectively). Increasing dietary roughage level linearly increased daily total chewing time; linearly elevated the mean, maximum, and minimum ruminal pH; and linearly decreased total VFA concentration and molar proportion of propionate in ruminal fluid. Saliva secretion during eating was increased, the secretion during rumination was unaffected, but the secretion during resting tended to decrease with increased dietary roughage level. As a result, total saliva secretion was not affected by treatments. In conclusion, the results of the present study did not support the concept that high-roughage diets elevated ruminal pH

  20. Adaptive responses of Bacillus cereus ATCC14579 cells upon exposure to acid conditions involve ATPase activity to maintain their internal pH.

    PubMed

    Senouci-Rezkallah, Khadidja; Jobin, Michel P; Schmitt, Philippe

    2015-03-05

    This study examined the involvement of ATPase activity in the acid tolerance response (ATR) of Bacillus cereus ATCC14579 strain. In the current work, B. cereus cells were grown in anaerobic chemostat culture at external pH (pHe ) 7.0 or 5.5 and at a growth rate of 0.2 h(-1) . Population reduction and internal pH (pHi ) after acid shock at pH 4.0 was examined either with or without ATPase inhibitor N,N'-dicyclohexylcarbodiimide (DCCD) and ionophores valinomycin and nigericin. Population reduction after acid shock at pH 4.0 was strongly limited in cells grown at pH 5.5 (acid-adapted cells) compared with cells grown at pH 7.0 (unadapted cells), indicating that B. cereus cells grown at low pHe were able to induce a significant ATR and Exercise-induced increase in ATPase activity. However, DCCD and ionophores had a negative effect on the ability of B. cereus cells to survive and maintain their pHi during acid shock. When acid shock was achieved after DCCD treatment, pHi was markedly dropped in unadapted and acid-adapted cells. The ATPase activity was also significantly inhibited by DCCD and ionophores in acid-adapted cells. Furthermore, transcriptional analysis revealed that atpB (ATP beta chain) transcripts was increased in acid-adapted cells compared to unadapted cells before and after acid shock. Our data demonstrate that B. cereus is able to induce an ATR during growth at low pH. These adaptations depend on the ATPase activity induction and pHi homeostasis. Our data demonstrate that the ATPase enzyme can be implicated in the cytoplasmic pH regulation and in acid tolerance of B. cereus acid-adapted cells.

  1. In Situ Observation of Active Oxygen Species in Fe-Containing Ni-Based Oxygen Evolution Catalysts: The Effect of pH on Electrochemical Activity.

    PubMed

    Trześniewski, Bartek J; Diaz-Morales, Oscar; Vermaas, David A; Longo, Alessandro; Bras, Wim; Koper, Marc T M; Smith, Wilson A

    2015-12-09

    Ni-based oxygen evolution catalysts (OECs) are cost-effective and very active materials that can be potentially used for efficient solar-to-fuel conversion process toward sustainable energy generation. We present a systematic spectroelectrochemical characterization of two Fe-containing Ni-based OECs, namely nickel borate (Ni(Fe)-B(i)) and nickel oxyhydroxide (Ni(Fe)OOH). Our Raman and X-ray absorption spectroscopy results show that both OECs are chemically similar, and that the borate anions do not play an apparent role in the catalytic process at pH 13. Furthermore, we show spectroscopic evidence for the generation of negatively charged sites in both OECs (NiOO(-)), which can be described as adsorbed "active oxygen". Our data conclusively links the OER activity of the Ni-based OECs with the generation of those sites on the surface of the OECs. The OER activity of both OECs is strongly pH dependent, which can be attributed to a deprotonation process of the Ni-based OECs, leading to the formation of the negatively charged surface sites that act as OER precursors. This work emphasizes the relevance of the electrolyte effect to obtain catalytically active phases in Ni-based OECs, in addition to the key role of the Fe impurities. This effect should be carefully considered in the development of Ni-based compounds meant to catalyze the OER at moderate pHs. Complementarily, UV-vis spectroscopy measurements show strong darkening of those catalysts in the catalytically active state. This coloration effect is directly related to the oxidation of nickel and can be an important factor limiting the efficiency of solar-driven devices utilizing Ni-based OECs.

  2. Accomplishments of the Abrupt Wing Stall (AWS) Program and Future Research Requirements

    NASA Technical Reports Server (NTRS)

    Hall, Robert M.; Woodson, Shawn H.; Chambers, Joseph R.

    2003-01-01

    The Abrupt Wing Stall (AWS) Program has addressed the problem of uncommanded lateral motions, such as wing drop and wing rock, at transonic speeds. The genesis of this Program was the experience of the F/A-1 8E/F Program in the late 1990's, when wing drop was discovered in the heart of the maneuver envelope for the pre-production aircraft. While the F/A-1 8E/F problem was subsequently corrected by a leading-edge flap scheduling change and the addition of a porous door to the wing fold fairing, the AWS Program was initiated as a national response to the lack of technology readiness available at the time of the F/A-18E/F Development Program. The AWS Program objectives were to define causal factors for the F/A-18E/F experience, to gain insights into the flow physics associated with wing drop, and to develop methods and analytical tools so that future programs could identify this type of problem before going to flight test. The paper reviews, for the major goals of the AWS Program, the status of the technology before the

  3. 14. Photocopy of a photographca. 1902taken by A.W. Peters showing ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    14. Photocopy of a photograph--ca. 1902--taken by A.W. Peters showing Eastwood and his party surveying for the Big Creek Project. The surveying party is visible in the upper right corner. Courtesy Mr. Charles Allan Whitney. - Little Rock Creek Dam, Little Rock Creek, Littlerock, Los Angeles County, CA

  4. Loss, Failure, and an Awful Reputation: A Response to Jonathan Silin

    ERIC Educational Resources Information Center

    Britt, Clare

    2013-01-01

    In this article the author responds to Jonathan Silin's article "At a Loss: Scared and Excited", and in doing so, takes up his ideas around the generative potential of loss. She uses these notions of loss to illuminate how, in one diverse school community in Australia, loss, failure and an "awful reputation" have opened up…

  5. Tank characterization report for double-shell tank 241-AW-102

    SciTech Connect

    Bell, K.E.

    1997-05-29

    This document summarizes the information on the historical uses, present status, and the sampling and analysis results of waste stored in Tank 241-AW-102. This report supports the requirements of the Tri-Party Agreement Milestone M-44-05.

  6. Supporting document for the historical tank content estimate for AW-tank farm

    SciTech Connect

    Brevick, C.H., Stroup, J.L.; Funk, J.W., Fluor Daniel Hanford

    1997-03-06

    This Supporting Document provides historical in-depth characterization information on AW-Tank Farm, such as historical waste transfer and level data, tank physical information, temperature plots, liquid observation well plots, chemical analyte and radionuclide inventories for the Historical Tank Content Estimate Report for the Southeast Quadrant of the Hanford 200 Areas.

  7. Ultrasonic Examination of Double-Shell Tank 241-AW-101 Examination Completed February 2009.

    SciTech Connect

    Pardini, Allan F.; Weier, Dennis R.; Anderson, Kevin K.

    2009-03-18

    AREVA Federal Services LLC (AFS), under a contract from Washington River Protection Solutions (WRPS), has performed an ultrasonic examination of selected portions of Double-Shell Tank 241-AW-101. PNNL is responsible for preparing a report(s) that describes the results of the AFS ultrasonic examinations.

  8. Ultrasonic Examination of Double-Shell Tank 241-AW-105 Examination Completed April 2009.

    SciTech Connect

    Pardini, Allan F.; Weier, Dennis R.; Anderson, Kevin K.; Hathaway, John E.

    2009-05-31

    AREVA Federal Services LLC (AFS), under a contract from Washington River Protection Solutions (WRPS), has performed an ultrasonic examination of selected portions of Double-Shell Tank 241-AW-105. PNNL is responsible for preparing a report(s) that describes the results of the AFS ultrasonic examinations.

  9. Ultrasonic Examination of Double-Shell Tank 241-AW-106 Examination Completed July 2009.

    SciTech Connect

    Pardini, Allan F.; Weier, Dennis R.; Anderson, Kevin K.; Hathaway, John E.

    2009-08-28

    AREVA Federal Services LLC (AFS), under a contract from Washington River Protection Solutions (WRPS), has performed an ultrasonic examination of selected portions of Double-Shell Tank 241-AW-106. PNNL is responsible for preparing a report(s) that describes the results of the AFS ultrasonic examinations.

  10. Influence of pH on the formulation of TiO2 powder prepared by co-precipitation of TiCl3 and photocatalytic activity

    NASA Astrophysics Data System (ADS)

    Yudoyono, Gatut; Zharvan, Vicran; Ichzan, Nur; Daniyati, Rizqa; Indarto, Bachtera; Pramono, Yono Hadi; Zainuri, Mochamad; Darminto

    2016-02-01

    Titanium dioxide (titania) nanoparticle were synthesized by coprecipitation process of titanium trichloride (TiCl3) in aqueous medium, with NH4OH as pH regulator. The pH solution was varied during the synthesis process between pH 3-8.4, and all samples were calcined temperature at 400°C for 3 hours. Characteristics and properties of the TiO2 powder were investigated using X-ray diffraction (XRD), Scanning Electron Microscopy (SEM). XRD results show that the single-phase rutile formed when the pH is less than 5, anatase single phase formed began pH 8, and the pH of the solution between 5-8 formed mixed phase rutile-anatase-brookite, rutile-brookite or anatase-brookite. Methylene Blue (MB) photodegradation test were performed in order to evaluate photocatalytic activity. Nanoparticles TiO2 rutile, anatase phase, and mixed phase rutile-brookite, anatase-brookite used to test the photocatalytic activity by measuring the absorbance spectrum photodegradation using UV-Vis spectrometer. The test results showed that the mixture phase of rutile-brookite provide the greatest photodegradation than other phases.

  11. Effects of inorganic nanoparticles on viability and catabolic activities of Agrobacterium sp. PH-08 during biodegradation of dibenzofuran.

    PubMed

    Le, Thao Thanh; Murugesan, Kumarasamy; Kim, Eun-Ju; Chang, Yoon-Seok

    2014-09-01

    This study investigated the cytotoxicity, genotoxicity, and growth inhibition effects of four different inorganic nanoparticles (NPs) such as aluminum (nAl), iron (nFe), nickel (nNi), and zinc (nZn) on a dibenzofuran (DF) degrading bacterium Agrobacterium sp. PH-08. NP (0-1,000 mg L(-1)) -treated bacterial cells were assessed for cytotoxicity, genotoxicity, growth and biodegradation activities at biochemical and molecular levels. In an aqueous system, the bacterial cells treated with nAl, nZn and nNi at 500 mg L(-1) showed significant reduction in cell viability (30-93.6 %, p < 0.05), while nFe had no significant inhibition on bacterial cell viability. In the presence of nAl, nZn and nNi, the cells exhibited elevated levels of reactive oxygen species (ROS), DNA damage and cell death. Furthermore, NP exposure showed significant (p < 0.05) impairment in DF and catechol biodegradation activities. The reduction in DF biodegradation was ranged about 71.7-91.6 % with single NPs treatments while reached up to 96.3 % with a mixture of NPs. Molecular and biochemical investigations also clearly revealed that NP exposure drastically affected the catechol-2,3-dioxygenase activities and its gene (c23o) expression. However, no significant inhibition was observed in nFe treatment. The bacterial extracellular polymeric materials and by-products from DF degradation can be assumed as key factors in diminishing the toxic effects of NPs, especially for nFe. This study clearly demonstrates the impact of single and mixed NPs on the microbial catabolism of xenobiotic-degrading bacteria at biochemical and molecular levels. This is the first study on estimating the impact of mixed NPs on microbial biodegradation.

  12. Influence of pH on the adsorption of uranium ions by oxidized activated carbon and chitosan

    SciTech Connect

    Park, G.I.; Park, H.S.; Woo, S.I.

    1999-03-01

    The adsorption characteristics of uranyl ions on surface-oxidized carbon were compared with those of powdered chitosan over a wide pH range. In particular, an extensive analysis was made on solution pH variation during the adsorption process or after adsorption equilibrium. Uranium adsorption on the two adsorbents was revealed to be strongly dependent on the initial pH of the solution. A quantitative comparison of the adsorption capacities of the two adsorbents was made, based on the isotherm data obtained at initial pH 3, 4, and 5. In order to analyze the adsorption kinetics incorporated with pH effects, batch experiments at various initial pH values were carried out, and solution pH profiles with the adsorption time were also evaluated. The breakthrough behavior in a column packed with oxidized carbon was also characterized with respect to the variation of effluent pH. Based on these experimental results, the practical applicability of oxidized carbon for uranium removal from acidic radioactive liquid waste was suggested.

  13. Somatic proembryo production from excised, wounded zygotic carrot embryos on hormone-free medium: evaluation of the effects of pH, ethylene and activated charcoal

    NASA Technical Reports Server (NTRS)

    Smith, D. L.; Krikorian, A. D.

    1990-01-01

    Wounded zygotic embryos of cultivated carrot produce somatic proembryos on hormone-free nutrient medium containing 1 mM NH4+ as the sole nitrogen source. Continued maintenance of proembryos on this medium leads to a "pure" culture of preglobular stage proembryos (PGSPs). Ethylene had no effect on this process. Also, somatic embryo production was not affected by growing cultures on activated charcoal-impregnated filter papers. However, somatic proembyros initiated on activated charcoal papers were not maintainable as PGSPs and developed into later embryo stages. Normally, medium pH dropped from 5.7 to 4 during each subculture period, but when using activated charcoal papers the pH endpoint was around 6 - 7 due to a leachable substance(s) within the filter papers. When powdered, activated charcoal was used in the medium as an adsorbent of products potentially released after wounding, pH dropped at the normal rate and to the expected levels; proembryos did not mature into later embryo stages and were maintainable exclusively as PGSPs. Low pH (approximately 4) is detrimental to proembyro production, but is essential to maintaining PGSPs on hormone-free nutrient medium, whereas a sustained pH > or = 5.7 allows continued development of PGSPs into later embryo stages.

  14. Effect of pH and temperature on the global compactness, structure, and activity of cellobiohydrolase Cel7A from Trichoderma harzianum.

    PubMed

    Colussi, Francieli; Garcia, Wanius; Rosseto, Flávio Rodolfo; de Mello, Bruno Luan Soares; de Oliveira Neto, Mário; Polikarpov, Igor

    2012-01-01

    Due to its elevated cellulolytic activity, the filamentous fungus Trichoderma harzianum (T. harzianum) has considerable potential in biomass hydrolysis application. Cellulases from Trichoderma reesei have been widely used in studies of cellulose breakdown. However, cellulases from T. harzianum are less-studied enzymes that have not been characterized biophysically and biochemically as yet. Here, we examined the effects of pH and temperature on the secondary and tertiary structures, compactness, and enzymatic activity of cellobiohydrolase Cel7A from T. harzianum (Th Cel7A) using a number of biophysical and biochemical techniques. Our results show that pH and temperature perturbations affect Th Cel7A stability by two different mechanisms. Variations in pH modify protonation of the enzyme residues, directly affecting its activity, while leading to structural destabilization only at extreme pH limits. Temperature, on the other hand, has direct influence on mobility, fold, and compactness of the enzyme, causing unfolding of Th Cel7A just above the optimum temperature limit. Finally, we demonstrated that incubation with cellobiose, the product of the reaction and a competitive inhibitor, significantly increased the thermal stability of Th Cel7A. Our studies might provide insights into understanding, at a molecular level, the interplay between structure and activity of Th Cel7A at different pH and temperature conditions.

  15. Inter and intrasite analyses of cultural materials from U20aw, Nye County, Nevada. Technical report No. 66

    SciTech Connect

    Hicks, P.A.; Pippin, L.C.; Henton, G.H.; Hemphill, M.L.; Lockett, C.L.

    1991-12-01

    In the Spring of 1986 Desert Research Institute (DRI) conducted a Class III archaeological survey of Drill Hole U20aw on the Nevada Test Site, Nye County, Nevada. Seven archaeological sites were located during the course of this survey including two temporary camps, four lithic scatters, and a possible pinyon cache. This report presents the results of the analyses of the data derived from all sites investigated during the data recovery operations on Drill Hole U20aw. Detailed analyses were focused on the spatial distribution of artifacts and features within and between sites in the southern portion of the study area (26Ny4867, 26Ny4869, and 26Ny4871). These analyses indicate that 26Ny4871 served principally as a temporary camp, while the area around the canyonhead to the east (which includes 26Ny4867 and 26Ny4869) seems to have been used as a site for both temporary camps and special activity loci. Projectile point styles suggest that the area was occupied from the Early Archaic into the early Historic period. Analyses of the artifacts that were recovered indicate that obsidian was the preferred material for all classes of flaked stone tools. All stages of lithic reduction are represented on the sites, but core reduction and thinning of bifaces appear to have been the primary activities. Processing of floral foods is indicated by the presence of several ground stone artifacts. Pinyon nuts and other items appear to have been stored in the area of 26Ny4869 and to the north of the drill hole as evidenced by the presence of several rock features that may have served as caches.

  16. Amino acid determinants conferring stable sialidase activity at low pH for H5N1 influenza A virus neuraminidase.

    PubMed

    Takahashi, Tadanobu; Nidom, Chairul A; Quynh Le, Mai Thi; Suzuki, Takashi; Kawaoka, Yoshihiro

    2012-01-01

    Avian influenza A viruses (IAVs) and human 1918, 1957, and 1968 pandemic IAVs all have neuraminidases (NAs) that are stable at low pH sialidase activity, yet most human epidemic IAVs do not. We examined the pH stability of H5N1 highly pathogenic avian IAV (HPAI) NAs and identified amino acids responsible for conferring stability at low pH. We found that, unlike other avian viruses, most H5N1 IAVs isolated since 2003 had NAs that were unstable at low pH, similar to human epidemic IAVs. These H5N1 viruses are thus already human virus-like and, therefore, have the frequent infections of humans.

  17. Evaporation of Pretreated Hanford Tank AW-101 Sample Mixed with Recycle

    SciTech Connect

    Crowder, M.L.

    2004-03-01

    This task was undertaken to provide validation data for the Treated Feed Evaporator flowsheet and operating conditions using Hanford Tank 241-AW-101 pretreated waste blended with simulated River Protection Project Waste Treatment Plant(RPP-WTP) waste recycles. This task provided the first opportunity to compare performance of blended actual treated waste to the performance of blended treated waste simulants and recycle simulants. Specific objectives were: (1) determining the effect of plant recycles; (2) developing solubility data, as well as chemical and physical property data; and (3) evaluating antifoam effectiveness. The treated AW-101 sample was produced from the composite effluent of Ion Exchange testing carried out at Savannah River Technical Center. The design basis operating conditions were validated. Blended solutions of pretreated Hanford Tank 241-AW-101 and simulated SBS A2 recycle boiled at expected RPP-WTP flowsheet conditions: 60 mm Hg and 50 degrees Celsius. As expected, boiling point increased with concentration. Specifically, the 35:65 blend of AW-101 to SBS A2 at 7.5 M sodium had boiling points of 53 degrees Celsius at 60 mm Hg and 59 degrees Celsius at 80 mm Hg. These were 6 to 7 degrees Celsius higher than those estimated by an OLI model. Precipitation of solids in the AW-101/SBS A2 matrix began between 8.7 M and 10.4 Msodium. No problematic or excessive solids were observed. Therefore, SBS recycles are not expected to have any adverse effect on the evaporation process in the RPP-WTP.

  18. SIRT3-AMPK Activation by Nitrite and Metformin Improves Hyperglycemia and Normalizes Pulmonary Hypertension Associated with Heart Failure with Preserved Ejection Fraction (PH-HFpEF)

    PubMed Central

    Lai, Yen-Chun; Tabima, Diana M.; Dube, John J.; Hughan, Kara S.; Vanderpool, Rebecca R.; Goncharov, Dmitry A.; St Croix, Claudette M.; Garcia-Ocaña, Adolfo; Goncharova, Elena A.; Tofovic, Stevan P.; Mora, Ana L.; Gladwin, Mark T.

    2016-01-01

    Background Pulmonary hypertension associated with heart failure with preserved ejection fraction (PH-HFpEF) is an increasingly recognized clinical complication of metabolic syndrome. No adequate animal model of PH-HFpEF is available and no effective therapies have been identified to date. A recent study suggested that dietary nitrate improves insulin resistance in eNOS null mice, and multiple studies have reported that both nitrate and its active metabolite, nitrite, have therapeutic activity in pre-clinical models of PH. Methods and Results In order to evaluate the efficacy and mechanism of nitrite in metabolic syndrome associated with PH-HFpEF, we developed a “two-hit” PH-HFpEF model in rats with multiple features of metabolic syndrome due to double leptin receptor defect (obese ZSF1) with the combined treatment of VEGF receptor blocker SU5416. Chronic oral nitrite treatment improved hyperglycemia in obese ZSF1 rats by a process that requires skeletal muscle SIRT3-AMPK-GLUT4 signaling. The glucose lowering effect of nitrite was abolished in SIRT3 deficient human skeletal muscle cells, as well as in SIRT3 knockout mice fed a high-fat diet. Skeletal muscle biopsies from humans with metabolic syndrome after 12 weeks of oral sodium nitrite and nitrate treatment (IND#115926) displayed increased activation of SIRT3 and AMPK. Finally, early treatments with nitrite and metformin at the time of SU5416 injection reduced pulmonary pressures and vascular remodeling in the PH-HFpEF model with robust activation of skeletal muscle SIRT3 and AMPK. Conclusions These studies validate a rodent model of metabolic syndrome and PH-HFpEF, suggesting a potential role of nitrite and metformin as a preventative treatment for this disease. PMID:26813102

  19. Luminal and Cytosolic pH Feedback on Proton Pump Activity and ATP Affinity of V-type ATPase from Arabidopsis*

    PubMed Central

    Rienmüller, Florian; Dreyer, Ingo; Schönknecht, Gerald; Schulz, Alexander; Schumacher, Karin; Nagy, Réka; Martinoia, Enrico; Marten, Irene; Hedrich, Rainer

    2012-01-01

    Proton pumping of the vacuolar-type H+-ATPase into the lumen of the central plant organelle generates a proton gradient of often 1–2 pH units or more. Although structural aspects of the V-type ATPase have been studied in great detail, the question of whether and how the proton pump action is controlled by the proton concentration on both sides of the membrane is not understood. Applying the patch clamp technique to isolated vacuoles from Arabidopsis mesophyll cells in the whole-vacuole mode, we studied the response of the V-ATPase to protons, voltage, and ATP. Current-voltage relationships at different luminal pH values indicated decreasing coupling ratios with acidification. A detailed study of ATP-dependent H+-pump currents at a variety of different pH conditions showed a complex regulation of V-ATPase activity by both cytosolic and vacuolar pH. At cytosolic pH 7.5, vacuolar pH changes had relative little effects. Yet, at cytosolic pH 5.5, a 100-fold increase in vacuolar proton concentration resulted in a 70-fold increase of the affinity for ATP binding on the cytosolic side. Changes in pH on either side of the membrane seem to be transferred by the V-ATPase to the other side. A mathematical model was developed that indicates a feedback of proton concentration on peak H+ current amplitude (vmax) and ATP consumption (Km) of the V-ATPase. It proposes that for efficient V-ATPase function dissociation of transported protons from the pump protein might become higher with increasing pH. This feature results in an optimization of H+ pumping by the V-ATPase according to existing H+ concentrations. PMID:22215665

  20. Effects of supplemental hay and corn silage versus full-time grazing on ruminal pH and chewing activity of dairy cows.

    PubMed

    Graf, C M; Kreuzer, M; Dohme, F

    2005-02-01

    Grazing young, highly digestible swards with and without supplemental hay or corn silage (5.5 kg of DM/d) offered overnight was tested for its effects on ruminal pH and chewing activity. A double 3 x 3 Latin square arrangement with 6 rumen-cannulated Brown Swiss cows (29 kg/d of milk) was applied. Herbage intake was quantified by controlled-release alkane capsules. Chewing activity was determined using an automatic microcomputer-based system for digital recording of the jaw movements. Except during milking, ruminal pH was measured continuously over 7 d by applying a device consisting of an indwelling pH electrode and a data-recording unit integrated in the cannula's cover. The grazing system had no significant effect on body weight, milk yield or composition (except milk urea), or total DM intake (13.5, 13.8, and 15.7 kg/d with full-time grazing, hay, and corn silage supplementation). No differences occurred for ruminating time per day and time per kilogram of DM intake. Full-time grazing cows spent more time eating per day (+26%) and time per kilogram of DM intake (+31%) than the other cows. Ruminal pH and time with pH <5.8 at night did not differ. Throughout the day, hay-supplemented cows had a significantly lower pH (-0.23) than full-time grazing cows, and the period of pH <5.8 was longer compared with corn-silage fed cows (77 vs. 11 min). Nocturnal supplement feeding gave no advantage over full-time grazing, and supplemental hay led to lower daytime pH.

  1. High sensitivity detection of cancer in vivo using a dual-controlled activation fluorescent imaging probe based on H-dimer formation and pH activation.

    PubMed

    Ogawa, Mikako; Kosaka, Nobuyuki; Regino, Celeste A S; Mitsunaga, Makoto; Choyke, Peter L; Kobayashi, Hisataka

    2010-05-01

    The key to improving the sensitivity of in vivo molecular imaging is to increase the target-to-background signal ratio (TBR). Optical imaging has a distinct advantage over other molecular imaging methods in that the fluorescent signal can be activated at the target thus reducing background signal. Previously, we found that H-dimer formation quenches fluorescence of xanthene fluorophores, and among these, TAMRA had the highest quenching ratio. Another approach to lowering background signal is to employ pH activation based on the photon-induced electron transfer (PeT) theory. We hypothesized that combining these two strategies could lead to greater quenching capacity than was possible with either probe alone. A pH-sensitive fluorophore, pHrodo or TAMRA was conjugated to the cancer targeting molecules, avidin (Av) and trastuzumab (Tra). As expected, both pHrodo and TAMRA formed H-dimers when conjugated to avidin or antibody and the dimerization resulted in efficient fluorescence quenching. In addition, pHrodo conjugated probes showed pH-dependent fluorescence activation. When the probes were used in an in vivo animal model, fluorescence endoscopy with Av-pHrodo depicted tumors with high TBR 1 h and 2 h after injection. Av-TAMRA also visualized tumors 1 h and 2 h after the injection, however, TBR was lower due to the background signal from non-specific binding 1 h after the injection as well as background fluorescence from the unbound agent. Thus, we demonstrate that a dual-controlled activatable optical probe based on the combination of H-dimer formation and pH activation can achieve high TBR at early time points during in vivo molecular imaging.

  2. Inclusion complexes of quercetin with three β-cyclodextrins derivatives at physiological pH: Spectroscopic study and antioxidant activity

    NASA Astrophysics Data System (ADS)

    Liu, Min; Dong, Lina; Chen, Aiju; Zheng, Yan; Sun, Dezhi; Wang, Xu; Wang, Bingquan

    2013-11-01

    Properties of the inclusion complexes of quercetin (QUE) with sulfobutyl ether-β-cyclodextrin (SBE-β-CD), hydroxypropyl-β-cyclodextrin (HP-β-CD), and methylated-β-cyclodextrin (M-β-CD) in tris-HCl buffer solutions of pH 7.40 were investigated. The stoichiometry and thermodynamic parameters for the complexation process (stability constants K, Gibbs free energy change ΔG, enthalpy change ΔH and entropy change ΔS) were determined using phase-solubility and fluorescence spectra analysis. The thermodynamic studies indicated that the inclusion reactions between QUE and the three β-CDs are enthalpy-driven processes. Proton nuclear magnetic resonance spectroscopy indicated that B-ring, C-ring, and part of A-ring of QUE interact with the cavity of β-CDs. The antioxidant activity of QUE and its inclusion complexes were determined by the scavenging of stable radical DPPH*. The results showed that the complexed QUE/CDs were more effective than free QUE, with the QUE/SBE-β-CD complex as the best form.

  3. Effect of temperature, pH, and water activity on Mucor spp. growth on synthetic medium, cheese analog and cheese.

    PubMed

    Morin-Sardin, Stéphanie; Rigalma, Karim; Coroller, Louis; Jany, Jean-Luc; Coton, Emmanuel

    2016-06-01

    The Mucor genus includes a large number of ubiquitous fungal species. In the dairy environment, some of them play a technological role providing typical organoleptic qualities to some cheeses while others can cause spoilage. In this study, we compared the effect of relevant abiotic factors for cheese production on the growth of six strains representative of dairy technological and contaminant species as well as of a non cheese related strain (plant endophyte). Growth kinetics were determined for each strain in function of temperature, water activity and pH on synthetic Potato Dextrose Agar (PDA), and secondary models were fitted to calculate the corresponding specific cardinal values. Using these values and growth kinetics acquired at 15 °C on cheese agar medium (CA) along with three different cheese types, optimal growth rates (μopt) were estimated and consequently used to establish a predictive model. Contrarily to contaminant strains, technological strains showed higher μopt on cheese matrices than on PDA. Interestingly, lag times of the endophyte strain were strongly extended on cheese related matrices. This study offers a relevant predictive model of growth that may be used for better cheese production control but also raises the question of adaptation of some Mucor strains to the cheese.

  4. Hepatic xanthine oxidase activity and purine nucleosides levels as physiological mediators to analyze a subcutaneous treatment with (PhSe)2 in mice infected by Toxoplasma gondii.

    PubMed

    Doleski, Pedro H; Leal, Daniela B R; Machado, Vanessa S; Bottari, Nathieli B; Casali, Emerson A; Moritz, Cesar E J; Camillo, Giovana; Vogel, Fernanda F; Stefani, Lenita M; da Silva, Aleksandro Schafer

    2017-03-01

    The aim of this study was to evaluate the levels of purine nucleosides and xanthine oxidase (XO) activity in the liver of mice chronically infected by Toxoplasma gondii and treated with diphenyl diselenide (PhSe)2. For this experiment, forty Swiss mice were used. Twenty animals were orally infected by approximately 50 bradizoites of a cystogenic ME-49 strain of T. gondii, and the same number of uninfected mice was used as a control group. Ten infected and ten uninfected mice were subcutaneously treated twice (days 1 and 20 post-infection (PI)) with 5 μmol kg(-1) of (PhSe)2. On day 30 PI, liver samples were collected to measure the levels of hypoxanthine (HYPO), xanthine (XAN), uric acid (UA), and XO activity. Infected animals showed increased (P < 0.05) levels of hepatic XAN and UA, as well as XO activity compared to uninfected animals. The use of (PhSe)2 in healthy mice increased the levels of all nucleosides, but decreased XO activity compared to healthy untreated animals. The group of infected and treated animals showed increased XAN and UA levels, and XO activity compared to the healthy control group, however infected and treated mice showed a decrease in the XO activity compared to the infected untreated group. We conclude that chronic infection caused by T. gondii can induce hepatic changes, such as increased UA levels and XO activity, that can increase the pro-oxidative profile. The (PhSe)2 treatment of healthy animals altered the levels of nucleosides, possibly due to low XO activity that decreased nucleoside degradation. Finally, (PhSe)2 treatment decreased XO activity in the infected group and increased nucleoside levels; however it was unable to reduce the UA levels found during the infection.

  5. Alterations in ROS Activity and Lysosomal pH Account for Distinct Patterns of Macroautophagy in LINCL and JNCL Fibroblasts

    PubMed Central

    Casanova, Bonaventura; Aguado, Carmen; Knecht, Erwin

    2013-01-01

    Neuronal Ceroid Lipofuscinoses (NCL) are lysosomal storage disorders characterized by the accumulation of lipofuscin within lysosomes. Late infantile (LINCL) and juvenile (JNCL) are their most common forms and are caused by loss-of-function mutations in tripeptidyl peptidase 1 (TPP1), a lysosomal endopeptidase, and CLN3 protein (CLN3p), whose location and function is still controversial. LINCL patients suffer more severely from NCL consequences than JNCL patients, in spite of having in common an abnormal accumulation of material with a similar composition in the lysosomes. To identify distinctive characteristics that could explain the differences in the severity of LINCL and JNCL pathologies, we compared the protein degradation mechanisms in patientś fibroblasts. Pulse-chase experiments show a significant decrease in protein degradation by macroautophagy in fibroblasts bearing TPP1 (CLN2) and CLN3p (CLN3) mutations. In CLN2 fibroblasts, LC3-II levels and other procedures indicate an impaired formation of autophagosomes, which confirms the pulse-chase experiments. This defect is linked to an accumulation of Reactive Oxygen Species (ROS), an upregulation of the Akt-mTOR signalling pathway and increased activities of the p38α and ERK1/2 MAPKs. In CLN3 fibroblasts, LC3-II analysis indicates impairment in autophagosome maturation and there is also a defect in fluid phase endocytosis, two alterations that can be related to an observed increase of 0.5 units in lysosomal pH. CLN3 fibroblasts also accumulate ROS but to a lower extent than CLN2. TPP1 activity is completely abrogated in CLN2 and partially diminished in CLN3 fibroblasts. TPP1 cleaves small hydrophobic proteins like subunit c of mitochondrial ATP synthase and the lack or a lower activity of this enzyme can contribute to lipofuscin accumulation. These alterations in TPP1 activity lead to an increased ROS production, especially in CLN2 in which it is aggravated by a decrease in catalase activity. This could

  6. Soil pH, total phosphorus, climate and distance are the major factors influencing microbial activity at a regional spatial scale

    NASA Astrophysics Data System (ADS)

    Cao, Haichuan; Chen, Ruirui; Wang, Libing; Jiang, Lanlan; Yang, Fen; Zheng, Shixue; Wang, Gejiao; Lin, Xiangui

    2016-05-01

    Considering the extensive functional redundancy in microbial communities and great difficulty in elucidating it based on taxonomic structure, studies on the biogeography of soil microbial activity at large spatial scale are as important as microbial community structure. Eighty-four soil samples were collected across a region from south to north China (about 1,000 km) to address the questions if microbial activity displays biogeographic patterns and what are driving forces. These samples represented different soil types, land use and climate. Redundancy analysis and nonmetric multidimensional scaling clearly revealed that soil microbial activities showed distinct differentiation at different sites over a regional spatial scale, which were strongly affected by soil pH, total P, rainfall, temperature, soil type and location. In addition, microbial community structure was greatly influenced by rainfall, location, temperature, soil pH and soil type and was correlated with microbial activity to some extent. Our results suggest that microbial activities display a clear geographic pattern that is greatly altered by geographic distance and reflected by climate, soil pH and total P over large spatial scales. There are common (distance, climate, pH and soil type) but differentiated aspects (TP, SOC and N) in the biogeography of soil microbial community structure and activity.

  7. Soil pH, total phosphorus, climate and distance are the major factors influencing microbial activity at a regional spatial scale

    PubMed Central

    Cao, Haichuan; Chen, Ruirui; Wang, Libing; Jiang, Lanlan; Yang, Fen; Zheng, Shixue; Wang, Gejiao; Lin, Xiangui

    2016-01-01

    Considering the extensive functional redundancy in microbial communities and great difficulty in elucidating it based on taxonomic structure, studies on the biogeography of soil microbial activity at large spatial scale are as important as microbial community structure. Eighty-four soil samples were collected across a region from south to north China (about 1,000 km) to address the questions if microbial activity displays biogeographic patterns and what are driving forces. These samples represented different soil types, land use and climate. Redundancy analysis and nonmetric multidimensional scaling clearly revealed that soil microbial activities showed distinct differentiation at different sites over a regional spatial scale, which were strongly affected by soil pH, total P, rainfall, temperature, soil type and location. In addition, microbial community structure was greatly influenced by rainfall, location, temperature, soil pH and soil type and was correlated with microbial activity to some extent. Our results suggest that microbial activities display a clear geographic pattern that is greatly altered by geographic distance and reflected by climate, soil pH and total P over large spatial scales. There are common (distance, climate, pH and soil type) but differentiated aspects (TP, SOC and N) in the biogeography of soil microbial community structure and activity. PMID:27170469

  8. Soil pH, total phosphorus, climate and distance are the major factors influencing microbial activity at a regional spatial scale.

    PubMed

    Cao, Haichuan; Chen, Ruirui; Wang, Libing; Jiang, Lanlan; Yang, Fen; Zheng, Shixue; Wang, Gejiao; Lin, Xiangui

    2016-05-12

    Considering the extensive functional redundancy in microbial communities and great difficulty in elucidating it based on taxonomic structure, studies on the biogeography of soil microbial activity at large spatial scale are as important as microbial community structure. Eighty-four soil samples were collected across a region from south to north China (about 1,000 km) to address the questions if microbial activity displays biogeographic patterns and what are driving forces. These samples represented different soil types, land use and climate. Redundancy analysis and nonmetric multidimensional scaling clearly revealed that soil microbial activities showed distinct differentiation at different sites over a regional spatial scale, which were strongly affected by soil pH, total P, rainfall, temperature, soil type and location. In addition, microbial community structure was greatly influenced by rainfall, location, temperature, soil pH and soil type and was correlated with microbial activity to some extent. Our results suggest that microbial activities display a clear geographic pattern that is greatly altered by geographic distance and reflected by climate, soil pH and total P over large spatial scales. There are common (distance, climate, pH and soil type) but differentiated aspects (TP, SOC and N) in the biogeography of soil microbial community structure and activity.

  9. Deletion of pH Regulator pac-3 Affects Cellulase and Xylanase Activity during Sugarcane Bagasse Degradation by Neurospora crassa

    PubMed Central

    Campos Antoniêto, Amanda Cristina; Ramos Pedersoli, Wellington; dos Santos Castro, Lílian; da Silva Santos, Rodrigo; Cruz, Aline Helena da Silva; Nogueira, Karoline Maria Vieira; Silva-Rocha, Rafael; Rossi, Antonio

    2017-01-01

    Microorganisms play a vital role in bioethanol production whose usage as fuel energy is increasing worldwide. The filamentous fungus Neurospora crassa synthesize and secrete the major enzymes involved in plant cell wall deconstruction. The production of cellulases and hemicellulases is known to be affected by the environmental pH; however, the regulatory mechanisms of this process are still poorly understood. In this study, we investigated the role of the pH regulator PAC-3 in N. crassa during their growth on sugarcane bagasse at different pH conditions. Our data indicate that secretion of cellulolytic enzymes is reduced in the mutant Δpac-3 at alkaline pH, whereas xylanases are positively regulated by PAC-3 in acidic (pH 5.0), neutral (pH 7.0), and alkaline (pH 10.0) medium. Gene expression profiles, evaluated by real-time qPCR, revealed that genes encoding cellulases and hemicellulases are also subject to PAC-3 control. Moreover, deletion of pac-3 affects the expression of transcription factor-encoding genes. Together, the results suggest that the regulation of holocellulase genes by PAC-3 can occur as directly as in indirect manner. Our study helps improve the understanding of holocellulolytic performance in response to PAC-3 and should thereby contribute to the better use of N. crassa in the biotechnology industry. PMID:28107376

  10. Deletion of pH Regulator pac-3 Affects Cellulase and Xylanase Activity during Sugarcane Bagasse Degradation by Neurospora crassa.

    PubMed

    Campos Antoniêto, Amanda Cristina; Ramos Pedersoli, Wellington; Dos Santos Castro, Lílian; da Silva Santos, Rodrigo; Cruz, Aline Helena da Silva; Nogueira, Karoline Maria Vieira; Silva-Rocha, Rafael; Rossi, Antonio; Silva, Roberto Nascimento

    2017-01-01

    Microorganisms play a vital role in bioethanol production whose usage as fuel energy is increasing worldwide. The filamentous fungus Neurospora crassa synthesize and secrete the major enzymes involved in plant cell wall deconstruction. The production of cellulases and hemicellulases is known to be affected by the environmental pH; however, the regulatory mechanisms of this process are still poorly understood. In this study, we investigated the role of the pH regulator PAC-3 in N. crassa during their growth on sugarcane bagasse at different pH conditions. Our data indicate that secretion of cellulolytic enzymes is reduced in the mutant Δpac-3 at alkaline pH, whereas xylanases are positively regulated by PAC-3 in acidic (pH 5.0), neutral (pH 7.0), and alkaline (pH 10.0) medium. Gene expression profiles, evaluated by real-time qPCR, revealed that genes encoding cellulases and hemicellulases are also subject to PAC-3 control. Moreover, deletion of pac-3 affects the expression of transcription factor-encoding genes. Together, the results suggest that the regulation of holocellulase genes by PAC-3 can occur as directly as in indirect manner. Our study helps improve the understanding of holocellulolytic performance in response to PAC-3 and should thereby contribute to the better use of N. crassa in the biotechnology industry.

  11. Effect of Carbon, Nitrogen Sources and Water Activity on Growth and Ochratoxin Production of Aspergillus carbonarius (Bainier) Thom

    PubMed Central

    Hashem, Abeer; Fathi Abd-Allah, Elsayed; Sultan Al-Obeed, Rashid; Abdullah Alqarawi, Abdulaziz; Alwathnani, Hend Awad

    2015-01-01

    Background: Ochratoxin A (OTA) is a toxic secondary metabolite produced by fungi belonging to Aspergillus and Penicillium genera. The production of OTA is influenced by environmental conditions and nutritional requirements. The postharvest application of bunches of table grape fruit (TGF), with water activity of 0.8 aw, was highly effective for controlling OTA contamination in vitro and in vivo (table grape). Objectives: The aim of this study was to determine the influence of environmental conditions and nutritional requirements on growth and OTA production by Aspergillus carbonarius, as well as, the impact of water activity on OTA production and growth characters of A. carbonarius. Furthermore, we also examined the influence of the application of different levels of water activity (aw 0.8) on the preservation of the general appearance of TGF and control of their contamination with OTA. Materials and Methods: The growth and OTA production by A. carbonarius were studied using glucose-ammonium nitrate salt broth medium. Effect of water activity was studied using glycerol (0.80, 0.85, 0.90, and 0.98 aw). The bunches of table grape fruits were immersed in glycerol solution (equivalent to 0.80 aw) and placed as a double layer in cardboard boxes (25 × 35 × 10 cm). The boxes were stored at 20°C for 15 days to simulate local market conditions. Results: The maximum OTA production by A. carbonarius was observed on broth medium after eight days of incubation at 20°C, with pH 4, and fructose and ammonium nitrate supplementation as carbon and nitrogen sources, respectively. The water activity (0.9, 0.85 aw) caused significant decrease in OTA production by A. carbonarius. The postharvest application of water activity (0.8 aw) was highly effective for maintenance of the table grape quality, which was expressed as weight loss, firmness and decay, while it also controlled OTA contamination of fruits under concept of local market conditions. Conclusions: Our results reported

  12. Universal dependence of hydrogen oxidation and evolution reaction activity of platinum-group metals on pH and hydrogen binding energy.

    PubMed

    Zheng, Jie; Sheng, Wenchao; Zhuang, Zhongbin; Xu, Bingjun; Yan, Yushan

    2016-03-01

    Understanding how pH affects the activity of hydrogen oxidation reaction (HOR) and hydrogen evolution reaction (HER) is key to developing active, stable, and affordable HOR/HER catalysts for hydroxide exchange membrane fuel cells and electrolyzers. A common linear correlation between hydrogen binding energy (HBE) and pH is observed for four supported platinum-group metal catalysts (Pt/C, Ir/C, Pd/C, and Rh/C) over a broad pH range (0 to 13), suggesting that the pH dependence of HBE is metal-independent. A universal correlation between exchange current density and HBE is also observed on the four metals, indicating that they may share the same elementary steps and rate-determining steps and that the HBE is the dominant descriptor for HOR/HER activities. The onset potential of CO stripping on the four metals decreases with pH, indicating a stronger OH adsorption, which provides evidence against the promoting effect of adsorbed OH on HOR/HER.

  13. Study of human serum albumin structure by dynamic light scattering: two types of reactions under different pH and interaction with physiologically active compounds

    NASA Astrophysics Data System (ADS)

    Luik, A. I.; Naboka, Yu. N.; Mogilevich, S. E.; Hushcha, T. O.; Mischenko, N. I.

    1998-09-01

    The effect of pH and binding of ten physiologically active compounds (isoproterenol, yohimbine, propranolol, clonidine, phenylephrine, carbachol, tripeptide fMLP, diphenhydramine, chlorpromazine and atropine) on the molecular structure of human serum albumin (HSA) has been studied using the dynamic light scattering. It was found that albumin globule has the most compact configuration (Stokes diameter 59-62 Å) at physiological pH 7.4. The changes in pH, both increase to 8.0 and decrease to 5.4, result in the growth of globule size to 72-81 Å. At acidic shift of pH an additional peak arises in the correlation spectra caused by the light scattering on the structures with the Stokes diameters of 29-37 Å. Those conform to the sizes of the albumin subdomains. The indicated peak is not displayed at basic shift of pH. The interaction with propranolol, clonidine, phenylephrine, carbachol and tripeptide fMLP which hinder adenylate cyclase (AdC) and activate Ca-polyphosphoinositide (Ca-PPI) signaling system of a cell initiates structural rearrangements similar to acidic transitions. Isoproterenol, yohimbine diphenhydramine, chlorpromazine and atropine, which activate AdC and hinder Ca-PPI, cause conformational changes of HSA similar to basic transitions.

  14. Effects of temperature, pH and NaCl on protease activity in digestive tract of young turbot, Scophthalmus maximus

    NASA Astrophysics Data System (ADS)

    Chen, Muyan; Zhang, Xiumei; Gao, Tianxiang; Chen, Chao

    2006-09-01

    The protease activity in digestive tract of young turbot Scophthalmus maximum was studied, and the optimal pH, temperature and NaCl concentration were determined for different portions of the fish's internal organs. The optimal activity in the fish's stomach was at pH of 2.2, while that in the intestinal extracts was within the alkaline range from 9.5 to 10.0. In hepatopancreas, the optimal pH was in low alkalinity at 8.5. The optimal reaction temperature was above 40°C in stomach, intestine and hepatopancreas. With increasing temperature, the pH value increased in stomach, while in the intestine, an opposite tendency was observed due to combined effect of pH and temperature. NaCl concentration showed inhibitory impact on protein digestion in hepatopancreas. The main protease for protein digestion in turbot seemed to be pepsin. Moreover, the maximum protease activity in different segments of intestine existed in the hindgut.

  15. Universal dependence of hydrogen oxidation and evolution reaction activity of platinum-group metals on pH and hydrogen binding energy

    PubMed Central

    Zheng, Jie; Sheng, Wenchao; Zhuang, Zhongbin; Xu, Bingjun; Yan, Yushan

    2016-01-01

    Understanding how pH affects the activity of hydrogen oxidation reaction (HOR) and hydrogen evolution reaction (HER) is key to developing active, stable, and affordable HOR/HER catalysts for hydroxide exchange membrane fuel cells and electrolyzers. A common linear correlation between hydrogen binding energy (HBE) and pH is observed for four supported platinum-group metal catalysts (Pt/C, Ir/C, Pd/C, and Rh/C) over a broad pH range (0 to 13), suggesting that the pH dependence of HBE is metal-independent. A universal correlation between exchange current density and HBE is also observed on the four metals, indicating that they may share the same elementary steps and rate-determining steps and that the HBE is the dominant descriptor for HOR/HER activities. The onset potential of CO stripping on the four metals decreases with pH, indicating a stronger OH adsorption, which provides evidence against the promoting effect of adsorbed OH on HOR/HER. PMID:27034988

  16. Effects of active and passive recovery on blood lactate and blood pH after a repeated sprint protocol in children and adults.

    PubMed

    Kappenstein, Jennifer; Engel, Florian; Fernández-Fernández, Jaime; Ferrauti, Alexander

    2015-02-01

    The aim of this study was to compare the effect of active (AR) and passive recovery (PR) after a high-intensive repeated sprint running protocol on physiological parameters in children and adults. Blood lactate (La) and blood pH were obtained during two sets of 5 × 5 s all-out sprints and several times during subsequent 30-min recovery in 16 children and 16 adults. End-exercise La was significantly lower and pH significantly higher in children (La: 5.21 ± 2.73 mmol·L-1; pH: 7.37 ± 0.06) compared with adults (La: 10.35 ± 5.76 mmol·L-1; pH: 7.27 ± 0.10) (p < .01). La half-life during postexercise recovery was significantly shorter in children (AR: 436 ± 371 s, PR: 830 ± 349 s) than in adults (AR: 733 ± 371 s, PR: 1361 ± 372 s), as well as in active compared with passive recovery for both age groups (p < .01). The age x recovery interaction for La half-life only approached statistical significance (p = .06). The results suggest a faster lactate disappearance and an earlier return to resting pH after a repeated sprint running protocol in children compared with adults and a less pronounced advantage of active recovery in children.

  17. Silver Nanoparticles Modified by Gelatin with Extraordinary pH Stability and Long-Term Antibacterial Activity

    PubMed Central

    Sivera, Martin; Kvitek, Libor; Soukupova, Jana; Panacek, Ales; Prucek, Robert; Vecerova, Renata; Zboril, Radek

    2014-01-01

    The potential for application of any nanoparticles, including silver nanoparticles (AgNPs), is strongly dependent on their stability against aggregation. Therefore, improvement of this parameter is a key task, especially in the case of AgNPs, because a correlation between size and biological activity has been demonstrated. In the present work, a natural stabilizer, gelatin, was investigated for the stabilization of AgNPs in an aqueous dispersion. The particles were prepared via a modified Tollens process, and the gelatin modifier was added prior to the reducing agent. The stability against aggregation of the AgNPs prepared by this method was more than one order of magnitude higher (on the basis of the critical coagulation concentration (CCC)) than that of AgNPs prepared via a similar method but without the assistance of gelatin. Their high stability against aggregation was confirmed over wide pH range (from 2 to 13) in which the particles did not exhibit rapid aggregation; such stability has not been previously reported for AgNPs. Additionally, gelatin not only fulfills the role of a unique stabilizer but also positively influences the modified Tollens process used to prepare the AgNPs. The diameter of the gelatin-modified AgNPs was substantially smaller in comparison to those prepared without gelatin. The polydispersity of the dispersion significantly narrowed. Moreover, the gelatin-stabilized AgNPs exhibited long-term stability against aggregation and maintained high antibacterial activity when stored for several months under ambient conditions. PMID:25098570

  18. The pH dependence of the cathodic peak potential of the active sites in bilirubin oxidase.

    PubMed

    Filip, Jaroslav; Tkac, Jan

    2014-04-01

    This is the first study showing pH dependence of three distinct redox sites within bilirubin oxidase (BOD) adsorbed on a nanocomposite modified electrode. The 1st redox centre with the highest redox potential Ec(1st)=404 mV vs. Ag/AgCl (614 mV vs. NHE at pH7.0) exhibited pH dependence with a slope -dEc(1st)/dpH=66(±3) mV under a non-turnover process. The 2nd redox centre with a potential Ec(2nd)=228 mV vs. Ag/AgCl (438 mV vs. NHE at pH7.0) was not dependent on pH in the absence and presence of O2. Finally, the 3rd redox site with a redox potential Ec(3rd)=92 mV vs. Ag/AgCl (302 mV vs. NHE at pH7.0) exhibited pH dependence for a cathodic process with -dEc(3rd)/dpH=70(±6) mV and for anodic process with -dEa(3rd)/dpH=73(±2) mV, respectively. Moreover, two break points for dependence of Ec(1st) or Ec(3rd) on pH were observed for the 1st (T1) site and the 3rd site assigned to involvement of two acidic amino acids (Asp105 and Glu463). A diagram of a potential difference between cathodic peaks of BOD as a dependence on pH is shown. The results obtained can be of interest for construction of biofuel cells based on BOD such as for generation of a low level of electricity from body fluids.

  19. Structure-Activity Analysis of Niclosamide Reveals Potential Role for Cytoplasmic pH in Control of Mammalian Target of Rapamycin Complex 1 (mTORC1) Signaling*

    PubMed Central

    Fonseca, Bruno D.; Diering, Graham H.; Bidinosti, Michael A.; Dalal, Kush; Alain, Tommy; Balgi, Aruna D.; Forestieri, Roberto; Nodwell, Matt; Rajadurai, Charles V.; Gunaratnam, Cynthia; Tee, Andrew R.; Duong, Franck; Andersen, Raymond J.; Orlowski, John; Numata, Masayuki; Sonenberg, Nahum; Roberge, Michel

    2012-01-01

    Mammalian target of rapamycin complex 1 (mTORC1) signaling is frequently dysregulated in cancer. Inhibition of mTORC1 is thus regarded as a promising strategy in the treatment of tumors with elevated mTORC1 activity. We have recently identified niclosamide (a Food and Drug Administration-approved antihelminthic drug) as an inhibitor of mTORC1 signaling. In the present study, we explored possible mechanisms by which niclosamide may inhibit mTORC1 signaling. We tested whether niclosamide interferes with signaling cascades upstream of mTORC1, the catalytic activity of mTOR, or mTORC1 assembly. We found that niclosamide does not impair PI3K/Akt signaling, nor does it inhibit mTORC1 kinase activity. We also found that niclosamide does not interfere with mTORC1 assembly. Previous studies in helminths suggest that niclosamide disrupts pH homeostasis of the parasite. This prompted us to investigate whether niclosamide affects the pH balance of cancer cells. Experiments in both breast cancer cells and cell-free systems demonstrated that niclosamide possesses protonophoric activity in cells and in vitro. In cells, niclosamide dissipated protons (down their concentration gradient) from lysosomes to the cytosol, effectively lowering cytoplasmic pH. Notably, analysis of five niclosamide analogs revealed that the structural features of niclosamide required for protonophoric activity are also essential for mTORC1 inhibition. Furthermore, lowering cytoplasmic pH by means other than niclosamide treatment (e.g. incubation with propionic acid or bicarbonate withdrawal) recapitulated the inhibitory effects of niclosamide on mTORC1 signaling, lending support to a possible role for cytoplasmic pH in the control of mTORC1. Our data illustrate a potential mechanism for chemical inhibition of mTORC1 signaling involving modulation of cytoplasmic pH. PMID:22474287

  20. Activation of Prodrug Treosulfan at pH 7.4 and 37°C Accompanied by Hydrolysis of Its Active Epoxides: Kinetic Studies with Clinical Relevance.

    PubMed

    Romański, Michał; Urbaniak, Bartosz; Kokot, Zenon; Główka, Franciszek K

    2015-12-01

    Treosulfan (TREO), originally registered for treatment of ovarian cancer, is currently being investigated for conditioning prior to hematopoietic stem cell transplantation. TREO is a prodrug, which undergoes a pH- and temperature-dependent two-step conversion to active monoepoxide [S,S-EBDM, (2S,3S)-1,2-epoxybutane-3,4-diol-4-methanesulfonate] and diepoxide [S,S-DEB, (2S,3S)-1,2:3,4-diepoxybutane]. In this paper, the kinetics of the nonenzymatic transformation of TREO at pH 7.4 and 37°C were studied for the first time including the effects of the TREO concentration, buffer concentration, ionic strength, and the presence of NaCl. Transformation of TREO was well described by a kinetic model, which included first-order reactions for TREO activation, that is, TREO → S,S-EBDM → S,S-DEB, and pseudo-first-order reactions for the hydrolytic decomposition of S,S-EBDM and S,S-DEB. In contrast to the two-step activation of TREO, the hydrolysis of epoxides was influenced by electrolytes. In phosphate-buffered saline, decomposition of S,S-EBDM and S,S-DEB (mean half-lives 25.7 and 15.4 h) proceeded much slower than their formation (mean half-lives 1.5 and 3.5 h). In conclusion, the kinetics of the nonenzymatic transformation of TREO in the presence of plasma electrolytes cannot contribute to the very low levels of S,S-EBDM and S,S-DEB observed in patient plasma. The results also indicate that elimination of TREO proceeds primarily via conversion to S,S-EBDM.

  1. Activated carbon enhanced ozonation of oxalate attributed to HO oxidation in bulk solution and surface oxidation: effect of activated carbon dosage and pH.

    PubMed

    Xing, Linlin; Xie, Yongbing; Minakata, Daisuke; Cao, Hongbin; Xiao, Jiadong; Zhang, Yi; Crittenden, John C

    2014-10-01

    Ozonation of oxalate in aqueous phase was performed with a commercial activated carbon (AC) in this work. The effect of AC dosage and solution pH on the contribution of hydroxyl radicals (HO) in bulk solution and oxidation on the AC surface to the removal of oxalate was studied. We found that the removal of oxalate was reduced by tert-butyl alcohol (tBA) with low dosages of AC, while it was hardly affected by tBA when the AC dosage was greater than 0.3g/L. tBA also inhibited ozone decomposition when the AC dosage was no more than 0.05g/L, but it did not work when the AC dosage was no less than 0.1g/L. These observations indicate that HO in bulk solution and oxidation on the AC surface both contribute to the removal of oxalate. HO oxidation in bulk solution is significant when the dosage of AC is low, whereas surface oxidation is dominant when the dosage of AC is high. The oxalate removal decreased with increasing pH of the solution with an AC dosage of 0.5g/L. The degradation of oxalate occurs mainly through surface oxidation in acid and neutral solution, but through HO oxidation in basic bulk solution. A mechanism involving both HO oxidation in bulk solution and surface oxidation was proposed for AC enhanced ozonation of oxalate.

  2. Alkaline Cytosolic pH and High Sodium Hydrogen Exchanger 1 (NHE1) Activity in Th9 Cells.

    PubMed

    Singh, Yogesh; Zhou, Yuetao; Shi, Xiaolong; Zhang, Shaqiu; Umbach, Anja T; Salker, Madhuri S; Lang, Karl S; Lang, Florian

    2016-11-04

    CD4(+) T helper 9 (Th9) cells are a newly discovered Th cell subset that produce the pleiotropic cytokine IL-9. Th9 cells can protect against tumors and provide resistance against helminth infections. Given their pivotal role in the adaptive immune system, understanding Th9 cell development and the regulation of IL-9 production could open novel immunotherapeutic opportunities. The Na(+)/H(+) exchanger 1 (NHE1; gene name Slc9α1)) is critically important for regulating intracellular pH (pHi), cell volume, migration, and cell survival. The pHi influences cytokine secretion, activities of membrane-associated enzymes, ion transport, and other effector signaling molecules such as ATP and Ca(2+) levels. However, whether NHE1 regulates Th9 cell development or IL-9 secretion has not yet been defined. The present study explored the role of NHE1 in Th9 cell development and function. Th cell subsets were characterized by flow cytometry and pHi was measured using 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein-acetoxymethyl ester (BCECF-AM) dye. NHE1 functional activity was estimated from the rate of realkalinization following an ammonium pulse. Surprisingly, in Th9 cells pHi and NHE1 activity were significantly higher than in all other Th cell subsets (Th1/Th2/Th17 and induced regulatory T cells (iTregs)). NHE1 transcript levels and protein abundance were significantly higher in Th9 cells than in other Th cell subsets. Inhibition of NHE1 by siRNA-NHE1 or with cariporide in Th9 cells down-regulated IL-9 and ATP production. NHE1 activity, Th9 cell development, and IL-9 production were further blunted by pharmacological inhibition of protein kinase Akt1/Akt2. Our findings reveal that Akt1/Akt2 control of NHE1 could be an important physiological regulator of Th9 cell differentiation, IL-9 secretion, and ATP production.

  3. Army Helicopter Night/Adverse Weather System (N/AWS)

    DTIC Science & Technology

    1976-06-11

    paper ^ that proved invaluable. My wife, Marty, not only +yped the many drafts and the final paper, but she also offered perceptive commeüts to...OPERATIONS Operation Feet (above ground Visibility level) Nighti Over flat terrain 500 1 mile Over mountainous terrain 1,000 above the...Fatigue has been appropriated by many branches of science as the designation for various regressive phenomena. With respect to human activity, fatigue

  4. Novel assay utilizing fluorochrome-tagged physostigmine (Ph-F) to in situ detect active acetylcholinesterase (AChE) induced during apoptosis.

    PubMed

    Huang, Xuan; Lee, Brian; Johnson, Gary; Naleway, John; Guzikowski, Anthony; Dai, Wei; Darzynkiewicz, Zbigniew

    2005-01-01

    It was recently reported that acetylcholinesterase (AChE) is expressed in cells undergoing apoptosis and that its presence is essential for assembly of the apoptosome and subsequent caspase-9 activation. To obtain a marker of active AChE that could assay this enzyme in live intact cells and be applicable to fluorescence microscopy and cytometry, the fluorescein-tagged physostigmine (Ph-F), high affinity ligand (inhibitor) reactive with the active center of AChE, was constructed and tested for its ability to in situ label AChE and measure its induction during apoptosis. Ph-F inhibited cholinesterase activity in vitro (IC50 = 10(-6) and 5 x 10(-6) M for equine butyrylcholinesterase and human erythrocyte AChE, respectively) and was a selective marker of cells and structures that were AChE-positive. Thus, exposure of mouse bone marrow cells to Ph-F resulted in the exclusive labeling of megakaryocytes, and of the diaphragm muscle, preferential labeling of the nerve-muscle junctions (end-plates). During apoptosis of carcinoma HeLa cells and leukemic HL-60 or Jurkat cells triggered either by the DNA topoisomerase 1 inhibitor topotecan (TPT) or by oxidative stress (H2O2), the cells become reactive with Ph-F. Their Ph-F derived fluorescence was measured by flow and laser scanning cytometry. The appearance of Ph-F binding sites during apoptosis was preceded by the loss of mitochondrial potential, was concurrent with the presence of activated caspases, and was followed by loss of membrane integrity. At a very early stage of apoptosis, when nucleolar segregation was apparent, the Ph-F binding sites were distinctly localized within the nucleolus and at later stages of apoptosis in the cytoplasm. During apoptosis triggered by TPT, Ph-F binding was preferentially induced in S-phase cells. Our data on megakaryocytes and end-plates indicate that Ph-F reacts with active sites of AChE, and can be used to reveal the presence of this enzyme in live cells and possibly to study its

  5. Catalytically active alkaline molten globular enzyme: Effect of pH and temperature on the structural integrity of 5-aminolevulinate synthase.

    PubMed

    Stojanovski, Bosko M; Breydo, Leonid; Hunter, Gregory A; Uversky, Vladimir N; Ferreira, Gloria C

    2014-12-01

    5-Aminolevulinate synthase (ALAS), a pyridoxal-5'phosphate (PLP)-dependent enzyme, catalyzes the first step of heme biosynthesis in mammals. Circular dichroism (CD) and fluorescence spectroscopies were used to examine the effects of pH (1.0-3.0 and 7.5-10.5) and temperature (20 and 37°C) on the structural integrity of ALAS. The secondary structure, as deduced from far-UV CD, is mostly resilient to pH and temperature changes. Partial unfolding was observed at pH2.0, but further decreasing pH resulted in acid-induced refolding of the secondary structure to nearly native levels. The tertiary structure rigidity, monitored by near-UV CD, is lost under acidic and specific alkaline conditions (pH10.5 and pH9.5/37°C), where ALAS populates a molten globule state. As the enzyme becomes less structured with increased alkalinity, the chiral environment of the internal aldimine is also modified, with a shift from a 420nm to 330nm dichroic band. Under acidic conditions, the PLP cofactor dissociates from ALAS. Reaction with 8-anilino-1-naphthalenesulfonic acid corroborates increased exposure of hydrophobic clusters in the alkaline and acidic molten globules, although the reaction is more pronounced with the latter. Furthermore, quenching the intrinsic fluorescence of ALAS with acrylamide at pH1.0 and 9.5 yielded subtly different dynamic quenching constants. The alkaline molten globule state of ALAS is catalytically active (pH9.5/37°C), although the kcat value is significantly decreased. Finally, the binding of 5-aminolevulinate restricts conformational fluctuations in the alkaline molten globule. Overall, our findings prove how the structural plasticity of ALAS contributes to reaching a functional enzyme.

  6. Role of different Escherichia coli hydrogenases in H+ efflux and F₁F(o)-ATPase activity during glycerol fermentation at different pH values.

    PubMed

    Blbulyan, Syuzanna; Avagyan, Arev; Poladyan, Anna; Trchounian, Armen

    2011-06-01

    Escherichia coli is able to ferment glycerol and produce H2 by different Hyds (hydrogenases). Wild-type whole cells were shown to extrude H+ through the F1Fo-ATPase and by other means with a lower rate compared with that under glucose fermentation. At pH 7.5, H+ efflux was stimulated in fhlA mutant (with defective transcriptional activator of Hyd-3 or Hyd-4) and was lowered in hyaB or hybC mutants (with defective Hyd-1 or Hyd-2) and hyaB hybC double mutant; DCCD (dicyclohexylcarbodi-imide)-sensitive H+ efflux was observed. At pH 5.5, H+ efflux in wild-type was lower compared with that at pH 7.5; it was increased in fhlA mutant and absent in hyaB hybC mutant. Membrane vesicle ATPase activity was lower in wild-type glycerol-fermented cells at pH 7.5 compared with that in glucose-fermented cells; 100 mM K+ did not stimulate ATPase activity. The latter at pH 7.5, compared with that in wild-type, was lower in hyaB and less in hybC mutants, stimulated in the hyaB hybC mutant and suppressed in the fhlA mutant; DCCD inhibited ATPase activity. At pH 5.5, the ATPase activities of hyaB and hybC mutants had similar values and were higher compared with that in wild-type; ATPase activity was suppressed in hyaB hybC and fhlA mutants. The results indicate that during glycerol fermentation, H+ was expelled also via F1Fo. At pH 7.5 Hyd-1 and Hyd-2 but not FhlA or Hyd-4 might be related to F1Fo or have their own H+-translocating ability. At pH 5.5, both Hyd-1 and Hyd-2 more than F1Fo might be involved in H+ efflux.

  7. Acidic digestion in a teleost: postprandial and circadian pattern of gastric pH, pepsin activity, and pepsinogen and proton pump mRNAs expression.

    PubMed

    Yúfera, Manuel; Moyano, Francisco J; Astola, Antonio; Pousão-Ferreira, Pedro; Martínez-Rodríguez, Gonzalo

    2012-01-01

    Two different modes for regulation of stomach acid secretion have been described in vertebrates. Some species exhibit a continuous acid secretion maintaining a low gastric pH during fasting. Others, as some teleosts, maintain a neutral gastric pH during fasting while the hydrochloric acid is released only after the ingestion of a meal. Those different patterns seem to be closely related to specific feeding habits. However, our recent observations suggest that this acidification pattern could be modified by changes in daily feeding frequency and time schedule. The aim of this study was to advance in understanding the regulation mechanisms of stomach digestion and pattern of acid secretion in teleost fish. We have examined the postprandial pattern of gastric pH, pepsin activity, and mRNA expression for pepsinogen and proton pump in white seabream juveniles maintained under a light/dark 12/12 hours cycle and receiving only one morning meal. The pepsin activity was analyzed according to the standard protocol buffering at pH 2 and using the actual pH measured in the stomach. The results show how the enzyme precursor is permanently available while the hydrochloric acid, which activates the zymogen fraction, is secreted just after the ingestion of food. Results also reveal that analytical protocol at pH 2 notably overestimates true pepsin activity in fish stomach. The expression of the mRNA encoding pepsinogen and proton pump exhibited almost parallel patterns, with notable increases during the darkness period and sharp decreases just before the morning meal. These results indicate that white seabream uses the resting hours for recovering the mRNA stock that will be quickly used during the feeding process. Our data clearly shows that both daily illumination pattern and feeding time are involved at different level in the regulation of the secretion of digestive juices.

  8. pH dependent catalytic activities of platinum nanoparticles with respect to the decomposition of hydrogen peroxide and scavenging of superoxide and singlet oxygen

    NASA Astrophysics Data System (ADS)

    Liu, Yi; Wu, Haohao; Li, Meng; Yin, Jun-Jie; Nie, Zhihong

    2014-09-01

    Recently, platinum (Pt) nanoparticles (NPs) have received increasing attention in the field of catalysis and medicine due to their excellent catalytic activity. To rationally design Pt NPs for these applications, it is crucial to understand the mechanisms underlying their catalytic and biological activities. This article describes a systematic study on the Pt NP-catalyzed decomposition of hydrogen peroxide (H2O2) and scavenging of superoxide (O2&z.rad;-) and singlet oxygen (1O2) over a physiologically relevant pH range of 1.12-10.96. We demonstrated that the catalytic activities of Pt NPs can be modulated by the pH value of the environment. Our results suggest that Pt NPs possess peroxidase-like activity of decomposing H2O2 into &z.rad;OH under acidic conditions, but catalase-like activity of producing H2O and O2 under neutral and alkaline conditions. In addition, Pt NPs exhibit significant superoxide dismutase-like activity of scavenging O2&z.rad;- under neutral conditions, but not under acidic conditions. The 1O2 scavenging ability of Pt NPs increases with the increase in the pH of the environment. The study will provide useful guidance for designing Pt NPs with desired catalytic and biological properties.Recently, platinum (Pt) nanoparticles (NPs) have received increasing attention in the field of catalysis and medicine due to their excellent catalytic activity. To rationally design Pt NPs for these applications, it is crucial to understand the mechanisms underlying their catalytic and biological activities. This article describes a systematic study on the Pt NP-catalyzed decomposition of hydrogen peroxide (H2O2) and scavenging of superoxide (O2&z.rad;-) and singlet oxygen (1O2) over a physiologically relevant pH range of 1.12-10.96. We demonstrated that the catalytic activities of Pt NPs can be modulated by the pH value of the environment. Our results suggest that Pt NPs possess peroxidase-like activity of decomposing H2O2 into &z.rad;OH under acidic conditions

  9. Respective Contribution of Mitochondrial Superoxide and pH to Mitochondria-targeted Circularly Permuted Yellow Fluorescent Protein (mt-cpYFP) Flash Activity*

    PubMed Central

    Wei-LaPierre, Lan; Gong, Guohua; Gerstner, Brent J.; Ducreux, Sylvie; Yule, David I.; Pouvreau, Sandrine; Wang, Xianhua; Sheu, Shey-Shing; Cheng, Heping; Dirksen, Robert T.; Wang, Wang

    2013-01-01

    Superoxide flashes are transient bursts of superoxide production within the mitochondrial matrix that are detected using the superoxide-sensitive biosensor, mitochondria-targeted circularly permuted YFP (mt-cpYFP). However, due to the pH sensitivity of mt-cpYFP, flashes were suggested to reflect transient events of mitochondrial alkalinization. Here, we simultaneously monitored flashes with mt-cpYFP and mitochondrial pH with carboxy-SNARF-1. In intact cardiac myocytes and purified skeletal muscle mitochondria, robust mt-cpYFP flashes were accompanied by only a modest increase in SNARF-1 ratio (corresponding to a pH increase of <0.1), indicating that matrix alkalinization is minimal during an mt-cpYFP flash. Individual flashes were also accompanied by stepwise increases of MitoSOX signal and decreases of NADH autofluorescence, supporting the superoxide origin of mt-cpYFP flashes. Transient matrix alkalinization induced by NH4Cl only minimally influenced flash frequency and failed to alter flash amplitude. However, matrix acidification modulated superoxide flash frequency in a bimodal manner. Low concentrations of nigericin (< 100 nm) that resulted in a mild dissipation of the mitochondrial pH gradient increased flash frequency, whereas a maximal concentration of nigericin (5 μm) collapsed the pH gradient and abolished flash activity. These results indicate that mt-cpYFP flash events reflect a burst in electron transport chain-dependent superoxide production that is coincident with a modest increase in matrix pH. Furthermore, flash activity depends strongly on a combination of mitochondrial oxidation and pH gradient. PMID:23457298

  10. Potentiation of anti-cancer drug activity at low intratumoral pH induced by the mitochondrial inhibitor m-iodobenzylguanidine (MIBG) and its analogue benzylguanidine (BG)

    PubMed Central

    Kuin, A; Aalders, M; Lamfers, M; Zuidam, D J van; Essers, M; Beijnen, J H; Smets, L A

    1999-01-01

    Tumour-selective acidification is of potential interest for enhanced therapeutic gain of pH sensitive drugs. In this study, we investigated the feasibility of a tumour-selective reduction of the extracellular and intracellular pH and their effect on the tumour response of selected anti-cancer drugs. In an in vitro L1210 leukaemic cell model, we confirmed enhanced cytotoxicity of chlorambucil at low extracellular pH conditions. In contrast, the alkylating drugs melphalan and cisplatin, and bioreductive agents mitomycin C and its derivative EO9, required low intracellular pH conditions for enhanced activation. Furthermore, a strong and pH-independent synergism was observed between the pH-equilibrating drug nigericin and melphalan, of which the mechanism is unclear. In radiation-induced fibrosarcoma (RIF-1) tumour-bearing mice, the extracellular pH was reduced by the mitochondrial inhibitor m-iodobenzylguanidine (MIBG) or its analogue benzylguanidine (BG) plus glucose. To simultaneously reduce the intracellular pH, MIBG plus glucose were combined with the ionophore nigericin or the Na+/H+ exchanger inhibitor amiloride and the Na+-dependent HCO3−/Cl−exchanger inhibitor 4,4′-diisothiocyanostilbene-2,2′-disulphonic acid (DIDS). Biochemical studies confirmed an effective reduction of the extracellular pH to approximately 6.2, and anti-tumour responses to the interventions indicated a simultaneous reduction of the intracellular pH below 6.6 for at least 3 h. Combined reduction of extra- and intracellular tumour pH with melphalan increased the tumour regrowth time to 200% of the pretreatment volume from 5.7 ± 0.6 days for melphalan alone to 8.1 ± 0.7 days with pH manipulation (P< 0.05). Mitomycin C related tumour growth delay was enhanced by the combined interventions from 3.8 ± 0.5 to 5.2 ± 0.5 days (P< 0.05), but only in tumours of relatively large sizes. The interventions were non-toxic alone or in combination with the anti-cancer drugs and did not affect

  11. A unique F-type H⁺-ATPase from Streptococcus mutans: an active H⁺ pump at acidic pH.

    PubMed

    Sasaki, Yuka; Nogami, Eri; Maeda, Masatomo; Nakanishi-Matsui, Mayumi; Iwamoto-Kihara, Atsuko

    2014-01-10

    We have shown previously that the Streptococcus mutans F-type H(+)-ATPase (F(O)F(1)) c subunit gene could complement Escherichia coli defective in the corresponding gene, particularly at acidic pH (Araki et al., (2013) [14]). In this study, the entire S. mutans F(O)F(1) was functionally assembled in the E. coli plasma membrane (SF(O)F(1)). Membrane SF(O)F(1) ATPase showed optimum activity at pH 7, essentially the same as that of the S. mutans, although the activity of E. coli F(O)F(1) (EF(O)F(1)) was optimum at pH≥9. The membranes showed detectable ATP-dependent H(+)-translocation at pH 5.5-6.5, but not at neutral conditions (pH≥7), consistent with the role of S. mutans F(O)F(1) to pump H(+) out of the acidic cytoplasm. A hybrid F(O)F(1), consisting of membrane-integrated F(O) and -peripheral F(1) sectors from S. mutans and E. coli (SF(O)EF(1)), respectively, essentially showed the same pH profile as that of EF(O)F(1) ATPase. However, ATP-driven H(+)-transport was similar to that by SF(O)F(1), with activity at acidic pH. Replacement of the conserved c subunit Glu53 in SF(O)F(1) abolished H(+)-transport at pH 6 or 7, suggesting its role in H(+) transport. Mutations in the SF(O)F(1) c subunit, Ser17Ala or Glu20Ile, changed the pH dependency of H(+)-transport, and the F(O) could transport H(+) at pH 7, as the membranes with EF(O)F(1). Ser17, Glu20, and their vicinity were suggested to be involved in H(+)-transport in S. mutans at acidic pH.

  12. Strategies to increase the stability of intermediate moisture foods towards Zygosaccharomyces rouxii: the effect of temperature, ethanol, pH and water activity, with or without the influence of organic acids.

    PubMed

    Vermeulen, A; Marvig, C L; Daelman, J; Xhaferi, R; Nielsen, D S; Devlieghere, F

    2015-02-01

    Intermediate moisture foods (IMF) are in general microbiologically stable products. However, due to health concerns consumer demands are increasingly forcing producers to lower the fat, sugar and preservatives content, which impede the stability of the IMF products. One of the strategies to counteract these problems is the storage of IMF products at lower temperatures. Thorough knowledge on growth/no growth boundaries of Zygosaccharomyces rouxii in IMF products, also at different storage temperatures is an important tool for ensuring microbiologically stability. In this study, growth/no growth models for Z. rouxii, developed by Vermeulen et al. (2012) were further extended by incorporating the factor temperature. Three different data sets were build: (i) without organic acids, (ii) with acetic acid (10,000 ppm on product basis) and (iii) with sorbic acid (1500 ppm on product basis). For each of these data sets three different growth/no growth models were developed after 30, 60 and 90 days. The results show that the influence of temperature is only significant in the lower temperature range (8-15 °C). Also, the effect of pH is negligible (pH 5.0-6.2) unless organic acids are present. More specific, acetic acid had only an additive effect to ethanol and aw at low pH, whereas sorbic acid had also an additive effect at the higher pH values. For incubation periods longer than 30 days the growth/no growth boundary remained stable but enlarged gradually between day 60 and 90, except for the lower temperature range (<12 °C) where the boundary shifts to more stringent environmental conditions.

  13. Effect of immobilization support, water activity, and enzyme ionization state on cutinase activity and enantioselectivity in organic media.

    PubMed

    Vidinha, Pedro; Harper, Neil; Micaelo, Nuno M; Lourenco, Nuno M T; da Silva, Marco D R Gomes; Cabral, Joaquim M S; Afonso, Carlos A M; Soares, Claudio M; Barreiros, Susana

    2004-02-20

    We studied the reaction between vinyl butyrate and 2-phenyl-1-propanol in acetonitrile catalyzed by Fusarium solani pisi cutinase immobilized on zeolites NaA and NaY and on Accurel PA-6. The choice of 2-phenyl-1-propanol was based on modeling studies that suggested moderate cutinase enantioselectivity towards this substrate. With all the supports, initial rates of transesterification were higher at a water activity (a(w)) of 0.2 than at a(w) = 0.7, and the reverse was true for initial rates of hydrolysis. By providing acid-base control in the medium through the use of solid-state buffers that control the parameter pH-pNa, which we monitored using an organo-soluble chromoionophoric indicator, we were able, in some cases, to completely eliminate dissolved butyric acid. However, none of the buffers used were able to improve the rates of transesterification relative to the blanks (no added buffer) when the enzyme was immobilized at an optimum pH of 8.5. When the enzyme was immobilized at pH 5 and exhibited only marginal activity, however, even a relatively acidic buffer with a pK(a) of 4.3 was able to restore catalytic activity to about 20% of that displayed for a pH of immobilization of 8.5, at otherwise identical conditions. As a(w) was increased from 0.2 to 0.7, rates of transesterification first increased slightly and then decreased. Rates of hydrolysis showed a steady increase in that a(w) range, and so did total initial reaction rates. The presence or absence of the buffers did not impact on the competition between transesterification and hydrolysis, regardless of whether the butyric acid formed remained as such in the reaction medium or was eliminated from the microenvironment of the enzyme through conversion into an insoluble salt. Cutinase enantioselectivity towards 2-phenyl-1-propanol was indeed low and was not affected by differences in immobilization support, enzyme protonation state, or a(w).

  14. Structural analysis of color video camera installation on tank 241AW101 (2 Volumes)

    SciTech Connect

    Strehlow, J.P.

    1994-08-24

    A video camera is planned to be installed on the radioactive storage tank 241AW101 at the DOE` s Hanford Site in Richland, Washington. The camera will occupy the 20 inch port of the Multiport Flange riser which is to be installed on riser 5B of the 241AW101 (3,5,10). The objective of the project reported herein was to perform a seismic analysis and evaluation of the structural components of the camera for a postulated Design Basis Earthquake (DBE) per the reference Structural Design Specification (SDS) document (6). The detail of supporting engineering calculations is documented in URS/Blume Calculation No. 66481-01-CA-03 (1).

  15. The effect of pH and chloride concentration on the stability and antimicrobial activity of chlorine-based sanitizers.

    PubMed

    Waters, Brian W; Hung, Yen-Con

    2014-04-01

    Chlorinated water and electrolyzed oxidizing (EO) water solutions were made to compare the free chlorine stability and microbicidal efficacy of chlorine-containing solutions with different properties. Reduction of Escherichia coli O157:H7 was greatest in fresh samples (approximately 9.0 log CFU/mL reduction). Chlorine loss in "aged" samples (samples left in open bottles) was greatest (approximately 40 mg/L free chlorine loss in 24 h) in low pH (approximately 2.5) and high chloride (Cl(-) ) concentrations (greater than 150 mg/L). Reduction of E. coli O157:H7 was also negatively impacted (<1.0 log CFU/mL reduction) in aged samples with a low pH and high Cl(-) . Higher pH values (approximately 6.0) did not appear to have a significant effect on free chlorine loss or numbers of surviving microbial cells when fresh and aged samples were compared. This study found chloride levels in the chlorinated and EO water solutions had a reduced effect on both free chlorine stability and its microbicidal efficacy in the low pH solutions. Greater concentrations of chloride in pH 2.5 samples resulted in decreased free chlorine stability and lower microbicidal efficacy.

  16. Air and Water System (AWS) Design and Technology Selection for the Vision for Space Exploration

    NASA Technical Reports Server (NTRS)

    Jones, Harry; Kliss, Mark

    2005-01-01

    This paper considers technology selection for the crew air and water recycling systems to be used in long duration human space exploration. The specific objectives are to identify the most probable air and water technologies for the vision for space exploration and to identify the alternate technologies that might be developed. The approach is to conduct a preliminary first cut systems engineering analysis, beginning with the Air and Water System (AWS) requirements and the system mass balance, and then define the functional architecture, review the International Space Station (ISS) technologies, and discuss alternate technologies. The life support requirements for air and water are well known. The results of the mass flow and mass balance analysis help define the system architectural concept. The AWS includes five subsystems: Oxygen Supply, Condensate Purification, Urine Purification, Hygiene Water Purification, and Clothes Wash Purification. AWS technologies have been evaluated in the life support design for ISS node 3, and in earlier space station design studies, in proposals for the upgrade or evolution of the space station, and in studies of potential lunar or Mars missions. The leading candidate technologies for the vision for space exploration are those planned for Node 3 of the ISS. The ISS life support was designed to utilize Space Station Freedom (SSF) hardware to the maximum extent possible. The SSF final technology selection process, criteria, and results are discussed. Would it be cost-effective for the vision for space exploration to develop alternate technology? This paper will examine this and other questions associated with AWS design and technology selection.

  17. Wounded Warriors (AW2), 5th Anniversary Report, 2004-2009

    DTIC Science & Technology

    2009-01-01

    Saffrons continue their commitment to the Army and wounded warriors. Recently SSG Saffron and his spouse, Colleen, accepted more than 400 holiday cards...from Harker Heights High School band students for the AW2 Soldiers at Fort Hood, TX, in December 2008. Colleen Saffron , who is extremely involved in...Additionally, Colleen Saffron is a founding partner for a nonprofit called Operation Life Transformed that provides funding to caregivers and spouses

  18. Initial Approaches to Mitigating the Impacts of the AWS-3 Auction (1755-1780 MHz)

    DTIC Science & Technology

    2015-05-13

    AWS-3, Aeronautical Mobile Telemetry, LTE-A, Transition Plan, Transmitters, Receivers, Antenna Systems, Bandwidth Efficient Modulation, Spectrum 16... antenna systems will be modified to desensitize them to commercial operations in the 1755-1780 MHz and 2155-2180 MHz bands • Better filters and LNA’s...Or TBD Technologies identified in upcoming studies • Telemetry antenna systems will be modified to provide bi- directional capability to control

  19. Cloud Computing for Pharmacometrics: Using AWS, NONMEM, PsN, Grid Engine, and Sonic.

    PubMed

    Sanduja, S; Jewell, P; Aron, E; Pharai, N

    2015-09-01

    Cloud computing allows pharmacometricians to access advanced hardware, network, and security resources available to expedite analysis and reporting. Cloud-based computing environments are available at a fraction of the time and effort when compared to traditional local datacenter-based solutions. This tutorial explains how to get started with building your own personal cloud computer cluster using Amazon Web Services (AWS), NONMEM, PsN, Grid Engine, and Sonic.

  20. Impact of automatization in temperature series in Spain and comparison with the POST-AWS dataset

    NASA Astrophysics Data System (ADS)

    Aguilar, Enric; López-Díaz, José Antonio; Prohom Duran, Marc; Gilabert, Alba; Luna Rico, Yolanda; Venema, Victor; Auchmann, Renate; Stepanek, Petr; Brandsma, Theo

    2016-04-01

    Climate data records are most of the times affected by inhomogeneities. Especially inhomogeneities introducing network-wide biases are sometimes related to changes happening almost simultaneously in an entire network. Relative homogenization is difficult in these cases, especially at the daily scale. A good example of this is the substitution of manual observations (MAN) by automatic weather stations (AWS). Parallel measurements (i.e. records taken at the same time with the old (MAN) and new (AWS) sensors can provide an idea of the bias introduced and help to evaluate the suitability of different correction approaches. We present here a quality controlled dataset compiled under the DAAMEC Project, comprising 46 stations across Spain and over 85,000 parallel measurements (AWS-MAN) of daily maximum and minimum temperature. We study the differences between both sensors and compare it with the available metadata to account for internal inhomogeneities. The differences between both systems vary much across stations, with patterns more related to their particular settings than to climatic/geographical reasons. The typical median biases (AWS-MAN) by station (comprised between the interquartile range) oscillate between -0.2°C and 0.4 in daily maximum temperature and between -0.4°C and 0.2°C in daily minimum temperature. These and other results are compared with a larger network, the Parallel Observations Scientific Team, a working group of the International Surface Temperatures Initiative (ISTI-POST) dataset, which comprises our stations, as well as others from different countries in America, Asia and Europe.

  1. Many particle approximation of the Aw-Rascle-Zhang second order model for vehicular traffic.

    PubMed

    Francesco, Marco Di; Fagioli, Simone; Rosini, Massimiliano D

    2017-02-01

    We consider the follow-the-leader approximation of the Aw-Rascle-Zhang (ARZ) model for traffic flow in a multi population formulation. We prove rigorous convergence to weak solutions of the ARZ system in the many particle limit in presence of vacuum. The result is based on uniform BV estimates on the discrete particle velocity. We complement our result with numerical simulations of the particle method compared with some exact solutions to the Riemann problem of the ARZ system.

  2. Determination of xanthine oxidoreductase activity in broilers: effect of pH and temperature of the assay and distribution in tissues.

    PubMed

    Carro, M D; Falkenstein, E; Blemings, K P; Klandorf, H

    2009-11-01

    Xanthine oxidoreductase (XOR) is the enzyme responsible for the synthesis of uric acid, which exists primarily in the dehydrogenase form in birds. Uric acid is the major end product of the metabolism of nitrogen-containing compounds in birds and it functions as an antioxidant to reduce oxidative stress. Despite the importance of this enzyme, the tissue distribution of XOR in physiologically normal chickens is not well known. In this study, we analyzed XOR activity in extracts of 8 tissues from broilers at 7 and 10 wk of age. No differences in XOR activity due to the age were found in any tissue. Liver and kidney showed the greatest activity, that in the kidney being about 89% of the activity in the liver. Enzyme activity in intestine and pancreas was about 60 and 37% of that in the liver. All breast muscle, heart, and lung samples showed enzyme activity, but values were only 3.0, 1.2, and 0.6% of those found in the liver. Traces of enzyme activity were also detected in 3 out of 10 brain samples, and no activity was found in the plasma. Our results show that XOR distribution in chickens differs from that in mammals, in which the highest levels have been found in liver and intestine. An additional objective was the evaluation of the effect of pH (7.2, 7.7, 8.2, and 8.7) and temperature (25 and 41 degrees C) on the enzyme activity in liver and kidney samples. Temperature had a similar effect on both tissues, with the activity at 25 degrees C being about 30% of that measured at 41 degrees C. At 41 degrees C, the enzyme activity in liver and kidney decreased quadratically as pH decreased from 8.7 to 7.2. The highest activity in kidney was measured at pH 8.2, although there were no differences between enzyme activities at pH 8.7 or 8.2 in the liver. Our results indicate that the optimum pH of the enzyme in chicken liver and kidney is around 8.2.

  3. Identification and description of the axillary web syndrome (AWS) by clinical signs, MRI and US imaging.

    PubMed

    Leduc, O; Fumière, E; Banse, S; Vandervorst, C; Clément, A; Parijs, T; Wilputte, F; Maquerlot, F; Ezquer Echandia, M; Tinlot, A; Leduc, A

    2014-12-01

    The Axillary Web Syndrome (AWS) follows surgery for breast neoplasia and consists of one, or more frequently two or three, cords of subcutaneous tissue. Cords originate from the axilla, spread to the antero-medial surface of the arm down to the elbow and then move into the antero-medial aspect of the forearm and sometimes into the root of the thumb. The purpose of this study was to compare two techniques, ultrasound (US) and Magnetic Resonance Imaging (MRI) for their sensitivity and accuracy in identifying AWS cords and to provide insights to the origin of this pathology. US examinations were performed on fifteen patients using a high frequency probe (17 MHz). We first palpated and marked the cord with location aided by maximum abduction. To identify the cord with MRI (1.5 Tesla), a catheter filled with a gel detectable under MRI was placed on the skin at the site of the cord. We found that in some US cases, the dynamic abduction maneuver was essential to facilitate detection of the cord. This dynamic method on ultrasound confirmed the precise location of the cord even if it was located deeper in the hypodermis fascia junction. US and MRI images revealed features of the cords and surrounding tissues. Imaging the cords was difficult with either of the imaging modalities. However, US seemed to be more efficient than MRI and allowed dynamic evaluation. Overall analysis of our study results supports a lymphatic origin of the AWS cord.

  4. Period studies and photometric models for two EB-type binaries EU Hya and AW Vul

    NASA Astrophysics Data System (ADS)

    Yang, Yuan-Gui; Li, Qun; Li, Hua-Li; Dai, Hai-Feng

    2016-02-01

    New photometry for two Algol-type binaries, EU Hya (P = 0.7782 d) and AW Vul (P = 0.8065 d), was carried out using the 60-cm telescope at Xinglong station of National Astronomical Observatories, Chinese Academy of Sciences. With the updated Wilson-Devinney code, photometric elements were derived from the multi-color light curves. The modeled results indicate that the two systems are near-contact binaries, whose secondary components fill their Roche lobes. The fill-out factors of the primaries are fp = 81.2 (±0.2)% for EU Hya and fp = 82.4 (±0.3)% for AW Vul. Period analysis implies that there exists a downward parabola with a light-time orbit from the (O - C) curve. This kind of periodic oscillation may be attributed to the light-time orbit effect of a third companion. The long-term period decrease may be caused by mass and angular momentum loss. When the orbital period decreases, the fill-out factor of fp will increase. Our results indicate that the primaries will also eventually fill their Roche lobes. EU Hya and AW Vul may possibly evolve from semi-detached binaries into contact ones.

  5. [Use of a gum elastic bougie for tracheal intubation with Pentax-AWS airway scope].

    PubMed

    Ueshima, Hironobu; Asai, Takashi; Shingu, Koh; Inoue, Hisashi; Hashimoto, Yuichi; Enomoto, Yoshiro; Okuda, Yasuhisa

    2008-01-01

    Pentax-AWS laryngoscope (Pentax, Tokyo, Japan) consists of a disposable anatomically shaped blade, a 12-cm cable with a charge-coupled device (CCD) camera and a 2.4-inch liquid crystal device (LCD) color monitor display. A tracheal tube can be attached to the right side of the blade. The device may be useful in patients with difficult airways. One limitation of the device is that intubation may be difficult if it is difficult to position the glottis to the target symbol on the monitor display. We experienced such a difficulty in four patients, and the use of a gum elastic bougie enabled intubation. In a 57-year-old woman with a difficult airway, tracheal intubation using either a Macintosh laryngoscope or a fiberscope had failed. By inserting the AWS laryngoscope, the glottis was easily seen on the monitor display. Nevertheless, it was difficult to position the glottis to the target symbol, and advancing a tracheal tube collided with the tissue around the glottis. A bougie was passed through the tracheal tube, and it became possible to insert the bougie into the trachea by adjusting the angle of its tip. The tracheal tube was then easily passed over the bougie into the trachea. We successfully used the same technique in other three patients. We believe that the gum elastic bougie can be useful for tracheal intubation using the Pentax-AWS laryngoscope.

  6. Cultural Heritage Protection Issues In Leśnica, The Settlement Of Wrocław

    NASA Astrophysics Data System (ADS)

    Kononowicz, Alena

    2015-09-01

    Leśnica, today the settlement on the western edge of Wrocław, formerly was an independent town, located on a previously wooded area, with a linear street system. It developed in the Middle Ages around the castle and church playing a service role for the Silesian Piast court on their way to Legnica and during hunting. In the thirteenth century it received city rights, and lost them in the eighteenth century. After the Piast dynasty had died out, it was sold by John of Luxembourg, and repeatedly changed its owners. In the nineteenth century it developed thanks to the industry, tourism and a convenient railway connection to Wrocław as well as hotel and restaurant facilities. In 1928, Leśnica was incorporated into Wrocław. After the Second World War, it lost its cultural continuity. In the 1970's, middle-heigh and high prefabricated buildings were built in the vicinity of a residential district. At the end of the twentieth and early twenty-first century, local industries were liquidated, and intensive land development started, causing the systematic blurring of its small-town character and its urban space started to acquire a character of a big city. In 2004, the old part of Leśnica was entered in the Register of Monuments. Also a ring road was planned, moving the cumbersome and dangerous transit traffic away from historic Średzka Street beyond the southern border of the settlement.

  7. Bioindication of heavy metals in the town Wrocław (Poland) with evergreen plants

    NASA Astrophysics Data System (ADS)

    Samecka-Cymerman, A.; Kempers, A. J.

    In this paper the results are reported of a study on the concentration of macroelements (N, P, K, Ca, Mg, S and Fe) and heavy metals (Ni, Cr, Co, V, Zn, Mn, Pb, Cd, Cu, Hg, Ba and Sr) in soil and in three species of the evergreen plants Ilex aquifolium, Mahonia aquifolium and Rhododendron catawbiense. These plants are collected in the Botanical Garden of the Polish town Wrocław (which is exposed to atmospheric exhausts of heavy traffic, chemical factories, metal smelters and a heat and power plant partly alimented with lignite and coals) and in two presumed unpolluted reference (background) sites, one in Poland (the Dendrological Garden in Wojslawice near Niemcza, south from Wrocław) and the other in The Netherlands (the Botanical Garden of the University of Nijmegen). Analyses of both soil and plant samples point to the pollution of the Wrocław Botanical Garden by Ni, Cr, Co, Cd via the atmosphere and pollution by Zn, Pb and Hg probably via atmosphere and soil. Especially pollution with Hg via soil is supported by a significant positive correlation between Hg content in soil and in all the examined species of which Ilex aquifolium seemed to be the best monitor of soil pollution with this element.

  8. The Listeria monocytogenes hemolysin has an acidic pH optimum to compartmentalize activity and prevent damage to infected host cells.

    PubMed

    Glomski, Ian J; Gedde, Margaret M; Tsang, Albert W; Swanson, Joel A; Portnoy, Daniel A

    2002-03-18

    Listeria monocytogenes is a facultative intracellular bacterial pathogen that escapes from a phagosome and grows in the host cell cytosol. The pore-forming cholesterol-dependent cytolysin, listeriolysin O (LLO), mediates bacterial escape from vesicles and is approximately 10-fold more active at an acidic than neutral pH. By swapping dissimilar residues from a pH-insensitive orthologue, perfringolysin O (PFO), we identified leucine 461 as unique to pathogenic Listeria and responsible for the acidic pH optimum of LLO. Conversion of leucine 461 to the threonine present in PFO increased the hemolytic activity of LLO almost 10-fold at a neutral pH. L. monocytogenes synthesizing LLO L461T, expressed from its endogenous site on the bacterial chromosome, resulted in a 100-fold virulence defect in the mouse listeriosis model. These bacteria escaped from acidic phagosomes and initially grew normally in cells and spread cell to cell, but prematurely permeabilized the host membrane and killed the cell. These data show that the acidic pH optimum of LLO results from an adaptive mutation that acts to limit cytolytic activity to acidic vesicles and prevent damage in the host cytosol, a strategy also used by host cells to compartmentalize lysosomal hydrolases.

  9. Identification of some factors affecting pharmaceutical active compounds (PhACs) removal in real wastewater. Case study of fungal treatment of reverse osmosis concentrate.

    PubMed

    Badia-Fabregat, Marina; Lucas, Daniel; Gros, Meritxell; Rodríguez-Mozaz, Sara; Barceló, Damià; Caminal, Glòria; Vicent, Teresa

    2015-01-01

    Many technologies are being developed for the efficient removal of micropollutants from wastewater and, among them, fungal degradation is one of the possible alternative biological treatments. In this article, some factors that might affect pharmaceutically active compounds (PhACs) removal in a fungal treatment of real wastewater were identified in batch bioreactor treating reverse osmosis concentrate (ROC) from urban wastewater treatment plant (WWTP). We found that degradation of PhACs by Trametes versicolor was enhanced by addition of external nutrients (global removal of 44%). Moreover, our results point out that high aeration might be involved in the increase in the concentration of some PhACs. In fact, conjugation and deconjugation processes (among others) affect the removal assessment of emerging contaminants when working with real concentrations in comparison to experiments with spiked samples. Moreover, factors that could affect the quantification of micropollutants at lab-scale experiments were studied.

  10. pH dependent catalytic activities of platinum nanoparticles with respect to the decomposition of hydrogen peroxide and scavenging of superoxide and singlet oxygen.

    PubMed

    Liu, Yi; Wu, Haohao; Li, Meng; Yin, Jun-Jie; Nie, Zhihong

    2014-10-21

    Recently, platinum (Pt) nanoparticles (NPs) have received increasing attention in the field of catalysis and medicine due to their excellent catalytic activity. To rationally design Pt NPs for these applications, it is crucial to understand the mechanisms underlying their catalytic and biological activities. This article describes a systematic study on the Pt NP-catalyzed decomposition of hydrogen peroxide (H2O2) and scavenging of superoxide (O2˙(-)) and singlet oxygen ((1)O2) over a physiologically relevant pH range of 1.12-10.96. We demonstrated that the catalytic activities of Pt NPs can be modulated by the pH value of the environment. Our results suggest that Pt NPs possess peroxidase-like activity of decomposing H2O2 into ˙OH under acidic conditions, but catalase-like activity of producing H2O and O2 under neutral and alkaline conditions. In addition, Pt NPs exhibit significant superoxide dismutase-like activity of scavenging O2˙(-) under neutral conditions, but not under acidic conditions. The (1)O2 scavenging ability of Pt NPs increases with the increase in the pH of the environment. The study will provide useful guidance for designing Pt NPs with desired catalytic and biological properties.

  11. The effect of linoleic acid on pH inside sodium bis(2-ethylhexyl)sulfosuccinate reverse micelles in isooctane and on the enzymic activity of soybean lipoxygenase.

    PubMed

    Rodakiewicz-Nowak, J; Maślakiewicz, P; Haber, J

    1996-06-01

    The effective pH of sodium bis(2-ethylhexyl)sulfosuccinate (AOT) reverse micelles (pHrm), containing buffers of different pH (pHst) and various amounts of linoleic acid, was studied within the range of compositions used to study the activity of soybean lipoxygenase in reverse micelles. Significant shifts of pHrm versus pHst were observed for the solutions of relatively higher pHst, dependent on linoleic acid and buffer concentrations. The effect diminished as pHst became closer to 7. When low-ionic-strength buffers were added to AOT solutions in isooctane, a significant buffering effect of linoleic acid in reverse micelles was observed. Solubilization of > 3 mM linoleic acid in micellar solutions containing 25 mM buffers gave the observed pHrm values almost independent of pHst. This effect diminished with the ionic strength of the buffering solution, but did not vanish even at 200 mM buffer. The observed effects result from the balance between ionization of linoleic acid and its partition between the water pool and the micellar interface. The enzymic activity of soybean lipoxygenase in the AOT reverse micellar solutions of the determined pHrm values was also studied. A significant reduction of the kinetics of the enzymic activity was observed, for all studied reverse micellar solutions. Changes of pHrm, caused by the presence of acidic substrate (linoleic acid) do not explain the observed reduction of activity directly through the effect on the enzyme. Due to unfavourable partition of the substrate between the microphases present in the systems, enhanced by reduction of pH at higher total concentrations of linoleic acid, the saturation of the enzyme with the substrate was not observed in the system and is difficult to attain experimentally in reverse micelles. A shift of the lipoxygenase activity/pHrm profile but negligible shift of the activity/pHst profile, with respect to aqueous buffer solutions, were observed. This indicates that either the information given

  12. Identifying Low pH Active and Lactate-Utilizing Taxa within Oral Microbiome Communities from Healthy Children Using Stable Isotope Probing Techniques

    PubMed Central

    McLean, Jeffrey S.; Fansler, Sarah J.; Majors, Paul D.; McAteer, Kathleen; Allen, Lisa Z.; Shirtliff, Mark E.; Lux, Renate; Shi, Wenyuan

    2012-01-01

    Background Many human microbial infectious diseases including dental caries are polymicrobial in nature. How these complex multi-species communities evolve from a healthy to a diseased state is not well understood. Although many health- or disease-associated oral bacteria have been characterized in vitro, their physiology within the complex oral microbiome is difficult to determine with current approaches. In addition, about half of these species remain uncultivated to date with little known besides their 16S rRNA sequence. Lacking culture-based physiological analyses, the functional roles of uncultivated species will remain enigmatic despite their apparent disease correlation. To start addressing these knowledge gaps, we applied a combination of Magnetic Resonance Spectroscopy (MRS) with RNA and DNA based Stable Isotope Probing (SIP) to oral plaque communities from healthy children for in vitro temporal monitoring of metabolites and identification of metabolically active and inactive bacterial species. Methodology/Principal Findings Supragingival plaque samples from caries-free children incubated with 13C-substrates under imposed healthy (buffered, pH 7) and diseased states (pH 5.5 and pH 4.5) produced lactate as the dominant organic acid from glucose metabolism. Rapid lactate utilization upon glucose depletion was observed under pH 7 conditions. SIP analyses revealed a number of genera containing cultured and uncultivated taxa with metabolic capabilities at pH 5.5. The diversity of active species decreased significantly at pH 4.5 and was dominated by Lactobacillus and Propionibacterium species, both of which have been previously found within carious lesions from children. Conclusions/Significance Our approach allowed for identification of species that metabolize carbohydrates under different pH conditions and supports the importance of Lactobacilli and Propionibacterium in the development of childhood caries. Identification of species within healthy subjects that

  13. Directed evolution and mutagenesis of glutamate decarboxylase from Lactobacillus brevis Lb85 to broaden the range of its activity toward a near-neutral pH.

    PubMed

    Shi, Feng; Xie, Yilong; Jiang, Junjun; Wang, Nannan; Li, Yongfu; Wang, Xiaoyuan

    2014-01-01

    Glutamate decarboxylase (GAD) transforms l-glutamate into γ-aminobutyric acid (GABA) with the consumption of a proton. GAD derived from lactic acid bacteria exhibits optimum activity at pH 4.0-5.0 and significantly loses activity at near-neutral pH. To broaden the active range of the GAD GadB1 from Lactobacillus brevis Lb85 toward a near-neutral pH, irrational design using directed evolution and rational design using site-specific mutagenesis were performed. For directed evolution of GadB1, a sensitive high-throughput screening strategy based on a pH indicator was established. One improved mutant, GadB1(T17I/D294G/Q346H), was selected from 800 variants after one round of EP-PCR. It exhibited 3.9- and 25.0-fold increase in activity and catalytic efficiency, respectively at pH 6.0. Through site-specific mutagenesis, several improved mutants were obtained, with GadB1(E312S) being the best one. The combined mutant GadB1(T17I/D294G/E312S/Q346H) showed even higher catalytic efficiency, 13.1- and 43.2-fold that of wild-type GadB1 at pH 4.6 and 6.0, respectively. The amount of GABA produced in gadB1(T17I/D294G/Q346H)-, gadB1(E312S)- and gadB1(T17I/D294G/E312S/Q346H)-expressing Corynebacterium glutamicum ATCC 13032 from endogenous l-glutamate increased by 9.6%, 20.3% and 63.9%, respectively. These results indicate that these mutations have beneficial effects on expanding the active pH range and on GABA biosynthesis, suggesting these GadB1 variants as potent candidates for GABA production.

  14. Determining salt concentrations for equivalent water activity in reduced-sodium cheese by use of a model system.

    PubMed

    Grummer, J; Schoenfuss, T C

    2011-09-01

    The range of sodium chloride (salt)-to-moisture ratio is critical in producing high-quality cheese products. The salt-to-moisture ratio has numerous effects on cheese quality, including controlling water activity (a(w)). Therefore, when attempting to decrease the sodium content of natural cheese it is important to calculate the amount of replacement salts necessary to create the same a(w) as the full-sodium target (when using the same cheese making procedure). Most attempts to decrease sodium using replacement salts have used concentrations too low to create the equivalent a(w) due to the differences in the molecular weight of the replacers compared with salt. This could be because of the desire to minimize off-flavors inherent in the replacement salts, but it complicates the ability to conclude that the replacement salts are the cause of off-flavors such as bitter. The objective of this study was to develop a model system that could be used to measure a(w) directly, without manufacturing cheese, to allow cheese makers to determine the salt and salt replacer concentrations needed to achieve the equivalent a(w) for their existing full-sodium control formulas. All-purpose flour, salt, and salt replacers (potassium chloride, modified potassium chloride, magnesium chloride, and calcium chloride) were blended with butter and water at concentrations that approximated the solids, fat, and moisture contents of typical Cheddar cheese. Salt and salt replacers were applied to the model systems at concentrations predicted by Raoult's law. The a(w) of the model samples was measured on a water activity meter, and concentrations were adjusted using Raoult's law if they differed from those of the full-sodium model. Based on the results determined using the model system, stirred-curd pilot-scale batches of reduced- and full-sodium Cheddar cheese were manufactured in duplicate. Water activity, pH, and gross composition were measured and evaluated statistically by linear mixed model

  15. 47 CFR 27.1176 - Cost-sharing requirements for AWS in the 2150-2160/62 MHz band.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... from this spectrum, including BRS incumbents occupying the 2150-2162 MHz band on a primary basis, must... § 27.1190. If an AWS licensee enters into a spectrum leasing arrangement and the spectrum...

  16. Anaerobic phosphate release from activated sludge with enhanced biological phosphorus removal. A possible mechanism of intracellular pH control

    SciTech Connect

    Bond, P.L.; Keller, J.; Blackall, L.L.

    1999-06-05

    The biochemical mechanisms of the wastewater treatment process known as enhanced biological phosphorus removal (EBPR) are presently described in a metabolic model. The authors investigated details of the EBPR model to determine the nature of the anaerobic phosphate release and how this may be metabolically associated with polyhydroxyalkanoate (PHA) formation. Iodoacetate, an inhibitor of glycolysis, was found to inhibit the anaerobic formation of PHA and phosphate release, supporting the pathways proposed in the EBPR metabolic model. In the metabolic model, it is proposed that polyphosphate degradation provides energy for the microorganisms in anaerobic regions of these treatment systems. Other investigations have shown that anaerobic phosphate release depends on the extracellular pH. The authors observed that when the intracellular pH of EBPR sludge was raised, substantial anaerobic phosphate release was caused without volatile fatty acid (VFA) uptake. Acidification of the sludge inhibited anaerobic phosphate release even in the presence of VFA. from these observations, the authors postulate that an additional possible role of anaerobic polyphosphate degradation in EBPR is for intracellular pH control. Intracellular pH control may be a metabolic feature of EBPR, not previously considered, that could have some use in the control and optimization of EBPR.

  17. The effect of pH on the exchangeability with deuterium of protons coupled to molybdenum(V) in the active and the desulpho forms of xanthine oxidase.

    PubMed Central

    Malthouse, J P; Bray, R C

    1983-01-01

    The effect of pH variation on the exchangeability with deuterium of protons strongly coupled to Mo(V) in the active and desulpho forms of xanthine oxidase was studied by e.p.r. and rapid freezing, in extension of the work of Gutteridge, Tanner & Bray [Biochem. J. (1978) 175, 887-897]. Above neutrality, exchange rates increased with increasing pH. Detailed studies were made on the desulpho enzyme under a variety of conditions, and exchange rate constants at 22 degrees C ranged from 0.16s -1 at pH 6.6 to 1.6s -1 at pH 11.3. The mechanism of proton exchange in the enzyme is discussed. The interpretation by the above workers that the strongly coupled proton of the active enzyme is on sulphur and that of the desulpho enzyme is on oxygen remains valid (and is in agreement with other work), as do their proposals for the structures of the protonated and deprotonated species. However, pK values cannot be calculated from the exchange data. It is likely that the relatively low rates of exchange observed are due to the difference of structure between the protonated and the deprotonated forms. In the case of the desulpho enzyme, an exchange mechanism, which involves the proton exchanging both as such and along with oxygen in the form of a hydroxyl ion, is discussed. PMID:6312970

  18. Effects of physically effective fiber on chewing activity and ruminal pH of dairy cows fed diets based on barley silage.

    PubMed

    Yang, W Z; Beauchemin, K A

    2006-01-01

    The objective of this study was to investigate the effects of physically effective neutral detergent fiber (peNDF) content of dairy cow diets containing barley silage as the sole forage source on feed intake, chewing activity, and ruminal pH. The experiment was designed as a replicated 3 x 3 Latin square using 6 lactating dairy cows with ruminal cannulas. Cows were offered 1 of 3 diets (high, medium, and low peNDF) obtained using barley silage that varied in particle length: long (theoretical cut length of 9.5 mm), medium (equal proportions of long and fine silages), and fine (theoretical cut length of 4.8 mm). The peNDF contents were determined using the Penn State Particle Separator and were 13.8, 11.8, and 10.5%, for the high, medium, and low diets, respectively. The physical effectiveness factors (defined as proportion retained on 19- and 8-mm screens) for the long and fine silages were 0.84 and 0.68, respectively. Increased forage particle size increased intake of peNDF but did not affect intake of DM and NDF. Ruminating and total chewing time were linearly increased with increasing dietary peNDF. Mean ruminal pH, area between the curve and a horizontal line drawn at pH 5.8 or 5.5, and time that pH was below 5.8 or 5.5 were not affected by peNDF content. Intake of peNDF was not correlated to any chewing activity but proportion of long particles on the 19-mm sieve tended to be correlated to ruminating chews (r = 0.36) and ruminating time (r = 0.36). These results indicate that increasing the peNDF content of diets increases chewing time. However, increased chewing time does not always improve ruminal pH status. Increasing chewing time and thus increasing salivary secretion may not fully overcome the effects of feed digestion and the production of fermentation acids that lower rumen pH. The results suggest that dietary peNDF and fermentable OM intake are critical in regulating rumen pH. Dietary particle size, expressed as peNDF, was a reliable indication of

  19. Influenza HA Subtypes Demonstrate Divergent Phenotypes for Cleavage Activation and pH of Fusion: Implications for Host Range and Adaptation

    PubMed Central

    Galloway, Summer E.; Reed, Mark L.; Russell, Charles J.; Steinhauer, David A.

    2013-01-01

    The influenza A virus (IAV) HA protein must be activated by host cells proteases in order to prime the molecule for fusion. Consequently, the availability of activating proteases and the susceptibility of HA to protease activity represents key factors in facilitating virus infection. As such, understanding the intricacies of HA cleavage by various proteases is necessary to derive insights into the emergence of pandemic viruses. To examine these properties, we generated a panel of HAs that are representative of the 16 HA subtypes that circulate in aquatic birds, as well as HAs representative of the subtypes that have infected the human population over the last century. We examined the susceptibility of the panel of HA proteins to trypsin, as well as human airway trypsin-like protease (HAT) and transmembrane protease, serine 2 (TMPRSS2). Additionally, we examined the pH at which these HAs mediated membrane fusion, as this property is related to the stability of the HA molecule and influences the capacity of influenza viruses to remain infectious in natural environments. Our results show that cleavage efficiency can vary significantly for individual HAs, depending on the protease, and that some HA subtypes display stringent selectivity for specific proteases as activators of fusion function. Additionally, we found that the pH of fusion varies by 0.7 pH units among the subtypes, and notably, we observed that the pH of fusion for most HAs from human isolates was lower than that observed from avian isolates of the same subtype. Overall, these data provide the first broad-spectrum analysis of cleavage-activation and membrane fusion characteristics for all of the IAV HA subtypes, and also show that there are substantial differences between the subtypes that may influence transmission among hosts and establishment in new species. PMID:23459660

  20. Modelling thermal inactivation of Listeria monocytogenes in sucrose solutions of various water activities.

    PubMed

    Fernández, A; López, M; Bernardo, A; Condón, S; Raso, J

    2007-06-01

    The heat resistance of Listeria monocytogenes was determined in sucrose solutions with water activity (a(w)) ranging from 0.99 to 0.90. At all temperatures investigated shape of the survival curves depended on the a(w) of the treatment medium. The survival curves for a(w)=0.99 appeared to be linear, for a(w)=0.96 were slightly upwardly concaved and for a(w)=0.93 and 0.90 were markedly concave upward. A mathematical model based on the Weibull distribution provided a good fit for all the survival curves obtained in this investigation. The effect of the temperature and a(w) on the Weibull model parameters was also studied. The shape parameter (p) depended on the a(w) of the treatment medium but in each medium of different a(w) the temperature did not have a significant effect on this parameter. The p parameter followed a linear relationship with a(w). The scale parameter (delta) decreased with the temperature following an exponential relationship and increased by decreasing the a(w) in the range from 0.99 to 0.93. However the delta parameter of survival curves obtained at a(w)=0.90 were lower than those obtained at a(w)=0.93. A mathematical model based on the Weibull parameters was built to describe the joint effect of temperature and a(w) on thermal inactivation of L. monocytogenes. This model provides a more complete information on the influence of the a(w) on the L. monocytogenes than the data initially generated. The model developed indicated that the effect of the a(w) on the thermal resistance of L. monocytogenes varied depending upon the temperature of treatment.

  1. Effects of Temperature and pH on the Activities of Catechol 2,3-dioxygenase Obtained from Crude Oil Contaminated Soil in Ilaje, Ondo State, Nigeria

    PubMed Central

    Olukunle, O.F.; Babajide, O.; Boboye, B.

    2015-01-01

    Enrichment technique was employed for the isolation of the crude oil degrading bacteria. The isolated bacteria were screened for their degradative ability and the best degrading bacteria were selected based on their growth. Specific activities of Catechol-2,3-dioxygenase and effects of temperature and pH and their stabilities on the enzyme relative activities were observed. Bacteria isolated from the soil sample include; Bacillus cereus, B. amyloliquficiens, B. firmus, Acinetobacter calcoaceticus, Pseudomonas sp. P. fluorescens, P.putida, P.aeruginosa, Achromobacter xylosoxidans and Achromobacter sp. Screening of the degradative ability of the bacteria revealed P. aeruginosa, Bacillus cereus, Acinetobacter calcoaceticus and Achromobacter sp. to be the best degraders. The pH and temperature range with time for the enzyme activity were 6.0-8.0 and 30oC-50oC respectively. The enzyme exhibited activity that was slightly more tolerant to alkaline pH. Therefore, engineering of Catechol 2,3-dioxygenase may be employed for application on bioremediation of polluted sites. PMID:26464607

  2. Modulation of rat and human cytochromes P450 involved in PhIP and 4-ABP activation by an aqueous extract of Phyllanthus orbicularis.

    PubMed

    Ferrer, Mirle; Cristófol, Carles; Sánchez-Lamar, Angel; Fuentes, Jorge Luís; Barbé, Jordi; Llagostera, Montserrat

    2004-02-01

    Phyllanthus orbicularis HBK (Euphorbiaceae) is a medicinal plant, endemic to Cuba, whose aqueous extract has proven antimutagenic effects against hydrogen peroxide and some promutagenic aromatic amines (AAs), in addition to its antiviral properties. In this paper, antimutagenesis of this extract against two carcinogenic AAs, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 4-aminobiphenyl (4-ABP) has been studied. Liver microsomal fractions from both induced rats and humans were used to metabolise both procarcinogenic compounds in the Salmonella assay. The plant extract was effective in reducing the mutagenesis of these AAs, activated by both kinds of fractions. The optimal antimutagenic effect was obtained when both AAs were metabolised by human enzymes, with an almost total reduction of 4-ABP mutagenesis and a decrease of about 75% of PhIP mutagenicity. Mutagenicity of both AAs, activated by induced rat fraction, was only decreased by about 50%. Inhibition by plant extract of alkoxyresorufin O-dealkylation activities, dependent on CYP1A, of both fractions was determined. In accordance with the results obtained, the inhibition or modulation of CYP1A subfamily activities, and possibly of CYP1A2, is thought to be the main mechanism of antimutagenesis of the aqueous extract of Phyllanthus orbicularis against 4-ABP and PhIP.

  3. Photoelectrochemical water oxidation by screen printed ZnO nanoparticle films: effect of pH on catalytic activity and stability

    NASA Astrophysics Data System (ADS)

    Fekete, Monika; Riedel, Wiebke; Patti, Antonio F.; Spiccia, Leone

    2014-06-01

    Nanostructured ZnO films are promising photoanode materials in photoelectrochemical water splitting. While such ZnO photoanodes have achieved high activity and good light conversion efficiency in the UV spectral region, their application in water splitting devices has been hampered by the susceptibility of ZnO towards photocorrosion in aqueous electrolytes. We report a systematic investigation aimed at optimising the electrolyte solution to improve the long-term stability of ZnO photoanodes. A stability diagram, based on the band edge positions of ZnO and the pH-dependent photodegradation potentials of ZnO (relative to the decomposition of water), indicates that the optimum pH operating conditions for ZnO photoanodes lie between pH 9-12.5. To verify this prediction experimentally, the activity and long-term stability of uniform screen-printed nano-ZnO films was tested in a wide range of buffered and non-buffered electrolytes (pH 6-13.5). The ZnO films were more active in buffered, than in non-buffered electrolytes, and the highest activities were observed close to the pKa of the phosphate and borate buffers used. Under zero applied potential, these screen-printed films achieved the highest reported photocurrents to date (0.42 mA cm-2 at pH 6 and 0.67 mA cm-2 at pH 10.5) for any pristine or modified ZnO-based water oxidation catalyst. The films were subjected to 12 h of controlled potential electrolysis, in selected electrolytes, under AM 1.5G simulated sunlight. The results are in good agreement with calculations based on thermodynamic data for ZnO. Films tested at pH 6 and 7 (representing typically used operating conditions) degraded rapidly, whereas they exhibited the highest stability when tested in a pH 10.5 borate buffer. In this case, 75% of the initial photoactivity was preserved after 12 hours, indicating that the lifetime of the electrode could be increased by over an order of magnitude compared to standard testing conditions.Nanostructured ZnO films are

  4. Glycolytic potential and activity of adenosine monophosphate kinase (AMPK), glycogen phosphorylase (GP) and glycogen debranching enzyme (GDE) in steer carcasses with normal (<5.8) or high (>5.9) 24h pH determined in M. longissimus dorsi.

    PubMed

    Apaoblaza, A; Galaz, A; Strobel, P; Ramírez-Reveco, A; Jeréz-Timaure, N; Gallo, C

    2015-03-01

    Muscle glycogen concentration (MGC) and lactate (LA), activity of glycogen debranching enzyme (GDE), glycogen phosphorylase (GP) and adenosine monophosphate kinase (AMPK) were determined at 0.5h (T0) and 24h (T24) post-mortem in Longissimus dorsi samples from 38 steers that produced high pH (>5.9) and normal pH (<5.8) carcasses at 24h postmortem. MGC, LA and glycolytic potential were higher (P<0.05) in normal pH carcasses. GDE activity was similar (P>0.05) in both pH categories. GP activity increased between T0 and T24 only in normal pH carcasses. AMPK activity was four times higher in normal pH v/s high pH carcasses, without changing its activity over time. Results reinforce the idea that differences in postmortem glycogenolytic/glycolytic flow in L. dorsi of steers showing normal v/s high muscle pH at 24h, could be explained not only by the higher initial MGC in normal pH carcasses, but also by a high and sustained activity of AMPK and an increased GP activity at 24h postmortem.

  5. Scanning electrochemical microscopy based evaluation of influence of pH on bioelectrochemical activity of yeast cells - Saccharomyces cerevisiae.

    PubMed

    Ramanavicius, A; Morkvenaite-Vilkonciene, I; Kisieliute, A; Petroniene, J; Ramanaviciene, A

    2017-01-01

    In this research scanning electrochemical microscopy was applied for the investigation of immobilized yeast Saccharomyces cerevisiae cells. Two redox mediators based system was applied in order to increase the efficiency of charge transfer from yeast cells. 9,10-phenanthrenequinone (PQ) was applied as a lipophilic redox mediator, which has the ability to cross the cell's membrane; another redox mediator was ferricyanide, which acted as a hydrophylic electron acceptor able to transfer electrons from the PQ to the working electrode of SECM. Hill's function was applied to determine the optimal pH for this described SECM-based system. The influence of pH on cell viability could be well described by Hill's function. It was determined that at pH 6.5 the PQ has a minimal toxic influence on yeast cells, and the kinetics of metabolic processes in cells as well as electron transfer rate achieved in consecutive action of both redox mediators were appropriate to achieve optimal current signals.

  6. Effects of ph ON Ni Coating on Poly(ethylene Terephthalate) Substrate by Printing Prime in Combination with Palladium Activating

    NASA Astrophysics Data System (ADS)

    Huang, Junjun; Sun, Zhiping; Huang, Hongzhi; Liu, Qi; Gao, Min; Li, Mengyu; Zhao, Feng; Chen, Zhenming

    2016-04-01

    In this work, the primer-printed and then self-assembled poly(ethylene terephthalate) (PET) sheets were plated when the bath pH increased from 7 to 11. The effects of bath pH on the structural and electrical properties of electroless nickel plating were investigated systematically using X-ray diffraction (XRD) and scanning electron microscopy (SEM). Results showed that the crystallographic structures of Ni-plated PET (NPP) sheets were face centered cubic, the preferential orientation of the coatings changes from (200) plane to (111) plane, the growth rate of plated nickel coating increased from 3.71μm/min to 8.13μm/min, the average Ni crystal size was increased from 0.23μm to 0.92μm, the average EMI-SE of NPP sheets was increased from 37.2dB to 38.6dB and the surface electrical resistivity of NPP sheets was decreased from 6235ohm ṡ cm to 0.03ohm ṡ cm with the increase of bath pH. The changes in structural and electrical properties were most possibly due to the fact that the thickness of coating increased.

  7. Study of the adsorption of Cd and Zn onto an activated carbon: Influence of pH, cation concentration, and adsorbent concentration

    SciTech Connect

    Seco, A.; Marzal, P.; Gabaldon, C.; Ferrer, J.

    1999-06-01

    The single adsorption of Cd and Zn from aqueous solutions has been investigated on Scharlau Ca 346 granular activated carbon in a wide range of experimental conditions: pH, metal concentration, and carbon concentration. The results showed the efficiency of the activated carbon as sorbent for both metals. Metal removals increase on raising the pH and carbon concentration, and decrease on raising the initial metal concentration. The adsorption processes have been modeled using the surface complex formation (SCF) Triple Layer Model (TLM). The adsorbent TLM parameters were determined. Modeling has been performed assuming a single surface bidentate species or an overall surface species with fractional stoichiometry. The bidentate stoichiometry successfully predicted cadmium and zinc removals in all the experimental conditions. The Freundlich isotherm has been also checked.

  8. Insights into the effect of soil pH on N(2)O and N(2) emissions and denitrifier community size and activity.

    PubMed

    Cuhel, Jirí; Simek, Miloslav; Laughlin, Ronnie J; Bru, David; Chèneby, Dominique; Watson, Catherine J; Philippot, Laurent

    2010-03-01

    The objective of this study was to investigate how changes in soil pH affect the N(2)O and N(2) emissions, denitrification activity, and size of a denitrifier community. We established a field experiment, situated in a grassland area, which consisted of three treatments which were repeatedly amended with a KOH solution (alkaline soil), an H(2)SO(4) solution (acidic soil), or water (natural pH soil) over 10 months. At the site, we determined field N(2)O and N(2) emissions using the (15)N gas flux method and collected soil samples for the measurement of potential denitrification activity and quantification of the size of the denitrifying community by quantitative PCR of the narG, napA, nirS, nirK, and nosZ denitrification genes. Overall, our results indicate that soil pH is of importance in determining the nature of denitrification end products. Thus, we found that the N(2)O/(N(2)O + N(2)) ratio increased with decreasing pH due to changes in the total denitrification activity, while no changes in N(2)O production were observed. Denitrification activity and N(2)O emissions measured under laboratory conditions were correlated with N fluxes in situ and therefore reflected treatment differences in the field. The size of the denitrifying community was uncoupled from in situ N fluxes, but potential denitrification was correlated with the count of NirS denitrifiers. Significant relationships were observed between nirS, napA, and narG gene copy numbers and the N(2)O/(N(2)O + N(2)) ratio, which are difficult to explain. However, this highlights the need for further studies combining analysis of denitrifier ecology and quantification of denitrification end products for a comprehensive understanding of the regulation of N fluxes by denitrification.

  9. Effect of growth temperature and pH on the aminopeptidase activity of Pseudomonas putida, P. fluorescens and Flavobacterium odoratum; the 4-nitroaniline test is reliable.

    PubMed

    Lopez-Tomas, L A; Ordoñez, J A; Mediavilla, C; Rodriguez-Marin, J L; Sarmiento, P; Zamora, A; Garcia de Fernando, G

    2008-01-01

    No significant difference (p > 0.05) was observed in the specific aminopeptidase activity (SAA) developed by Pseudomonas fluorescens, P. putida and Flavobacterium odoratum either growing at pH 5.0-6.5 or at 7 and 12 degrees C. Nevertheless, a significant difference was found when comparing the SAA of these organisms. The SAA of F. odoratum was lower than those of pseudomonads. The 4-nitroaniline test is reliable to estimate the G(-) load of fresh food products.

  10. Hydrolysis and volatile fatty acids accumulation of waste activated sludge enhanced by the combined use of nitrite and alkaline pH.

    PubMed

    Huang, Cheng; Liu, Congcong; Sun, Xiuyun; Sun, Yinglu; Li, Rui; Li, Jiansheng; Shen, Jinyou; Han, Weiqing; Liu, Xiaodong; Wang, Lianjun

    2015-12-01

    Volatile fatty acids (VFAs) production from anaerobic digestion of waste activated sludge (WAS) is often limited by the slow hydrolysis and/or poor substrate availability. Increased attention has been given to enhance the hydrolysis and acidification of WAS recently. This study presented an efficient and green strategy based on the combined use of nitrite pretreatment and alkaline pH to stimulate hydrolysis and VFA accumulation from WAS. Results showed that both proteins and polysaccharides increased in the presence of nitrite, indicating the enhancement of sludge solubilization and hydrolysis processes. Mechanism investigations showed that nitrite pretreatment could disintegrate the sludge particle and disperse extracellular polymeric substances (EPS). Then, anaerobic digestion tests demonstrated VFA production increased with nitrite treatment. The maximal VFA accumulation was achieved with 0.1 g N/L nitrite dosage and pH 10.0 at a sludge retention time (SRT) of 7 days, which was much higher VFA production in comparison with the blank, sole nitrite pretreatment, or sole pH 10. The potential analysis suggested that the combined nitrite pretreatment and alkaline pH is capable of enhancing WAS digestion with a great benefit for biological nutrient removal (BNR).

  11. Effect of muscle and post-mortem rate of pH and temperature fall on antioxidant enzyme activities in beef.

    PubMed

    Pastsart, Umaporn; De Boever, Maarten; Claeys, Erik; De Smet, Stefaan

    2013-03-01

    The aim of this study was to investigate the effect of muscle, inner and outer Musculus biceps femoris (IBF and OBF respectively) and Musculus longissimus dorsi (LD), on the post-mortem rate of pH and temperature fall, and the activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) during simulated retail display. At day 0 of display (2 days post-mortem), the CAT and GSH-Px activities were lower in IBF than in OBF and LD (P<0.001), and the SOD activity was lower in OBF compared to IBF and LD (P<0.001). At day 10 of display, SOD and CAT activities had decreased in all three muscles compared to day 0 (P<0.001), whereas the GSH-Px activity did increase with time of display. Across muscles, there were significant relationships between temperature fall, colour, lipid and colour stability and antioxidant enzyme activities.

  12. Inter and intrasite analyses of cultural materials from U20aw, Nye County, Nevada. Technical report No. 66

    SciTech Connect

    Hicks, P.A.; Pippin, L.C.; Henton, G.H.

    1991-12-01

    This archaeological presents the results of the analyses of the data derived from all sites investigated during the data recovery operations on Drill Hole U20aw on the Nevada Test Site, Nye County Nevada. These sites were determined to be potentially eligible for inclusion in the national Register of Historic Places. Detailed analyses were focused on the spatial distribution of artifacts and features within and between sites in the southern portion of the study area. These analyses indicate that one area served principally as a temporary camp, while the area around the canyonhead to the east seems to have been used as a site for both temporary camps and special activity loci. Projectile point styles suggest that the area was occupied from the Early Archaic into the early Historic period. Analyses of the artifacts that were recovered indicate that obsidian was the preferred material for all classes of flaked stone tools. All stages of lithic reduction are represented on the sites, but core reduction and thinning of bifaces appear to have been the primary activities. Processing of floral foods is indicated by the presence of several ground stone artifacts. Pinyon nuts and other items appear to have been stored in several areas as evidenced by the presence of several rock features that may have served as caches.

  13. ON ABSORPTION BY CIRCUMSTELLAR DUST, WITH THE PROGENITOR OF SN 2012aw AS A CASE STUDY

    SciTech Connect

    Kochanek, C. S.; Khan, R.; Dai, X.

    2012-11-01

    We use the progenitor of SN 2012aw to illustrate the consequences of modeling circumstellar dust using Galactic (interstellar) extinction laws that (1) ignore dust emission in the near-IR and beyond, (2) average over dust compositions, and (3) mischaracterize the optical/UV absorption by assuming that scattered photons are lost to the observer. The primary consequences for the progenitor of SN 2012aw are that both the luminosity and the absorption are significantly overestimated. In particular, the stellar luminosity is most likely in the range 10{sup 4.8} < L {sub *}/L {sub Sun} < 10{sup 5.0} and the star was not extremely massive for a Type IIP progenitor, with M {sub *} < 15 M {sub Sun }. Given the properties of the circumstellar dust and the early X-ray/radio detections of SN 2012aw, the star was probably obscured by an ongoing wind with M-dot {approx}10{sup -5.5} to 10{sup -5.0} M {sub Sun} yr{sup -1} at the time of the explosion, roughly consistent with the expected mass-loss rates for a star of its temperature (T{sub *} {approx_equal} 3600{sup +300} {sub -200} K) and luminosity. In the spirit of Galactic extinction laws, we supply simple interpolation formulae for circumstellar extinction by dusty graphitic and silicate shells as a function of wavelength ({lambda} {>=} 0.3 {mu}m) and total (absorption plus scattering) V-band optical depth ({tau}{sub V} {<=} 20). These do not include the contributions of dust emission, but provide a simple, physical alternative to incorrectly using interstellar extinction laws.

  14. Analysis of the Foundation Slab Settlement of the "Africa Pavilion" Facility in Wrocław

    NASA Astrophysics Data System (ADS)

    Sawicki, Eugeniusz; Strzelecki, Tomasz

    2015-12-01

    This article presents the results of numerical calculations of soil consolidation underneath the "Africa Pavilion" structure in Wrocław Zoo, Poland. To determine the deformations of the baseplate of the "Africa Pavilion" and deformations of the subsoil, Biot's consolidation theory for two-phase medium was applied. The calculations were carried out using the professional program FlexPDE v.6, which is based on the Finite Element Method. Numerical calculations performed were used to evaluate the design assumptions allowing for the laying of hydraulic conduits under the slab.

  15. Boildown Study on Supernatant Liquid Retrieved from AW-106 in December 2012

    SciTech Connect

    Page, Jason S.

    2013-06-04

    This document reports the results of a boildown study using a composite created from supernatant liquid grab samples retrieved from tank 241-AW-106 in December of 2012. The composite was made using predetermined volumes of the grab samples which accounted for layering of the supernatant liquid in the tank. The finished composite was a clear, yellow liquid containing no visible solids at hot cell ambient temperatures (24 - 27 °C). The density of the test composite was measured in the hot cell immediately before the boildown study and was 1.266 g/mL at 27.1 °C.

  16. Extensions and amplifications of a traffic model of Aw and Rascle

    SciTech Connect

    Greenberg, J. M.

    2000-08-01

    In a recent paper [1] Aw and Rascle introduced a new model of traffic on a uni-directional highway. Here the author studies an extension of this model, one which accounts for drivers attempting to travel at their maximum allowable speed. The author looks at a Lagrangian reformulation of this problem; a formulation that leads to an effective computational algorithm for solving the resulting system. He also investigates approximation scheme introduced by Dafermos [5] for scalar conservation laws and demonstrates that this Dafermos scheme works well on this 2 x 2 system.

  17. An Overview of the Components of AW-IPM Campaigns against the New World Screwworm

    PubMed Central

    Mastrangelo, Thiago; Welch, John B.

    2012-01-01

    The New World Screwworm, Cochliomyia hominivorax (Coquerel), is one of the most damaging parasites of livestock, causing millions of dollars in annual losses to producers. The fly is an obligate parasite of warm-blooded animals, including humans. After a successful 50-year eradication campaign, C. hominivorax has been eradicated from the USA, Mexico and Central America by an area-wide integrated pest management approach. Recently, Caribbean and South American countries have expressed an interest in this approach. Aiming to support forthcoming projects in these countries, this review describes the main technical components of past and ongoing AW-IPM campaigns against C. hominivorax. PMID:26466720

  18. A Cumulative Spore Killing Approach: Synergistic Sporicidal Activity of Dilute Peracetic Acid and Ethanol at Low pH Against Clostridium difficile and Bacillus subtilis Spores.

    PubMed

    Nerandzic, Michelle M; Sankar C, Thriveen; Setlow, Peter; Donskey, Curtis J

    2016-01-01

    Background.  Alcohol-based hand sanitizers are the primary method of hand hygiene in healthcare settings, but they lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We previously demonstrated that acidification of ethanol induced rapid sporicidal activity, resulting in ethanol formulations with pH 1.5-2 that were as effective as soap and water washing in reducing levels of C difficile spores on hands. We hypothesized that the addition of dilute peracetic acid (PAA) to acidified ethanol would enhance sporicidal activity while allowing elevation of the pH to a level likely to be well tolerated on skin (ie, >3). Methods.  We tested the efficacy of acidified ethanol solutions alone or in combination with PAA against C difficile and Bacillus subtilis spores in vitro and against nontoxigenic C difficile spores on hands of volunteers. Results.  Acidification of ethanol induced rapid sporicidal activity against C difficile and to a lesser extent B subtilis. The addition of dilute PAA to acidified ethanol resulted in synergistic enhancement of sporicidal activity in a dose-dependent fashion in vitro. On hands, the addition of 1200-2000 ppm PAA enhanced the effectiveness of acidified ethanol formulations, resulting in formulations with pH >3 that were as effective as soap and water washing. Conclusions.  Acidification and the addition of dilute PAA induced rapid sporicidal activity in ethanol. Our findings suggest that it may be feasible to develop effective sporicidal ethanol formulations that are safe and tolerable on skin.

  19. A Cumulative Spore Killing Approach: Synergistic Sporicidal Activity of Dilute Peracetic Acid and Ethanol at Low pH Against Clostridium difficile and Bacillus subtilis Spores

    PubMed Central

    Nerandzic, Michelle M.; Sankar C, Thriveen; Setlow, Peter; Donskey, Curtis J.

    2016-01-01

    Background. Alcohol-based hand sanitizers are the primary method of hand hygiene in healthcare settings, but they lack activity against bacterial spores produced by pathogens such as Clostridium difficile and Bacillus anthracis. We previously demonstrated that acidification of ethanol induced rapid sporicidal activity, resulting in ethanol formulations with pH 1.5–2 that were as effective as soap and water washing in reducing levels of C difficile spores on hands. We hypothesized that the addition of dilute peracetic acid (PAA) to acidified ethanol would enhance sporicidal activity while allowing elevation of the pH to a level likely to be well tolerated on skin (ie, >3). Methods. We tested the efficacy of acidified ethanol solutions alone or in combination with PAA against C difficile and Bacillus subtilis spores in vitro and against nontoxigenic C difficile spores on hands of volunteers. Results. Acidification of ethanol induced rapid sporicidal activity against C difficile and to a lesser extent B subtilis. The addition of dilute PAA to acidified ethanol resulted in synergistic enhancement of sporicidal activity in a dose-dependent fashion in vitro. On hands, the addition of 1200–2000 ppm PAA enhanced the effectiveness of acidified ethanol formulations, resulting in formulations with pH >3 that were as effective as soap and water washing. Conclusions. Acidification and the addition of dilute PAA induced rapid sporicidal activity in ethanol. Our findings suggest that it may be feasible to develop effective sporicidal ethanol formulations that are safe and tolerable on skin. PMID:26885539

  20. [Influences of pH and complexing agents on degradation of reactive brilliant blue KN-R by ferrous activated persulfate].

    PubMed

    Zhang, Cheng; Wan, Jin-Quan; Ma, Yong-Wen; Wang, Yan; Huang, Ming-Zhi; Lan, Ming

    2012-03-01

    The influences of pH and complexing agents on degradation of reactive brilliant blue KN-R by ferrous activated persulfate were investigated, and finally the complexed and the uncomplexed system were compared. Because the lower the pH the more quickly will be the dissociation of S2O8(2-) to the SO4(-*) and the ORP of the dominant radical SO4(-*) in the acidic condition is higher than the dominant radical *OH in the alkaline condition, KN-R degradation rates in acidic condition are far outweigh in the neutral and alkaline conditions. When pH value was 3, the residual rate of KN-R was 17.0% within 3 hours by EDTA system with the lowest PS consumption rate 32.3%, so EDTA was the best complexing agent choice in acidic condition. When pH value was 7, the residual rates of KN-R were 11.3%, 12.4% within 3 days by EDTA, citric acid system with the PS residual rates 28.9%, 28.0% respectively, so EDTA, citric acid were the better choices in the neutral condition. When pH value was 10, glucose acid, citric acid, EDTA and tartaric acid systems all had the similar KN-R degradation rates and PS residual rates, so all could acted as the complexing agents, when the system contained trace amounts of ferrous, the addition of complexing agent would greatly improve the degradation rate of pollutant, from original 52.5% to 79.3% of 3 d, so PS is suitable for in situ chemical oxidation (ISCO).

  1. Energy separations for the electronic states of PH -2,PH 2 and PH +2

    NASA Astrophysics Data System (ADS)

    Balasubramanian, K.

    1993-03-01

    All-electron complete-active space multi-configuration self-consistent field (CASSCF) followed by second-order configuration interaction (SOCI) calculations in conjunction with large P(13s10p3d2flg/7s6p3d2flg) and H (10s5p1d/8s5p1d) basis sets are made on the electronic states of PH -2, PH 2 and PH +2. We compute the adiabatic electron affinities of PH 2 and PH. The 3B 1-X 1A 1, 1B 1-X 1A 1 energy separations of PH +2 and the 2A 1-X 2B 1 energy separation of PH 2 are computed.

  2. A silica-supported iron oxide catalyst capable of activating hydrogen peroxide at neutral pH values.

    PubMed

    Pham, Anh Le-Tuan; Lee, Changha; Doyle, Fiona M; Sedlak, David L

    2009-12-01

    Iron oxides catalyze the conversion of hydrogen peroxide (H(2)O(2)) into oxidants capable of transforming recalcitrant contaminants. Unfortunately, the process is relatively inefficient at circumneutral pH values because of competing reactions that decompose H(2)O(2) without producing oxidants. Silica- and alumina-containing iron oxides prepared by sol-gel processing of aqueous solutions containing Fe(ClO(4))(3), AlCl(3), and tetraethyl orthosilicate efficiently catalyzed the decomposition of H(2)O(2) into oxidants capable of transforming phenol at circumneutral pH values. Relative to hematite, goethite, and amorphous FeOOH, the silica-iron oxide catalyst exhibited a stoichiometric efficiency, defined as the number of moles of phenol transformed per mole of H(2)O(2) consumed, which was 10-40 times higher than that of the iron oxides. The silica-alumina-iron oxide catalyst had a stoichiometric efficiency that was 50-80 times higher than that of the iron oxides. The significant enhancement in oxidant production is attributable to the interaction of Fe with Al and Si in the mixed oxides, which alters the surface redox processes, favoring the production of strong oxidants during H(2)O(2) decomposition.

  3. Anti-Leukemia Activity of In Vitro-Expanded Human Gamma Delta T Cells in a Xenogeneic Ph+ Leukemia Model

    PubMed Central

    Siegers, Gabrielle M.; Felizardo, Tania C.; Mathieson, A. Mark; Kosaka, Yoko; Wang, Xing-Hua; Medin, Jeffrey A.; Keating, Armand

    2011-01-01

    Gamma delta T cells (GDTc) lyse a variety of hematological and solid tumour cells in vitro and in vivo, and are thus promising candidates for cellular immunotherapy. We have developed a protocol to expand human GDTc in vitro, yielding highly cytotoxic Vgamma9/Vdelta2 CD27/CD45RA double negative effector memory cells. These cells express CD16, CD45RO, CD56, CD95 and NKG2D. Flow cytometric, clonogenic, and chromium release assays confirmed their specific cytotoxicity against Ph+ cell lines in vitro. We have generated a fluorescent and bioluminescent Ph+ cell line, EM-2eGFPluc, and established a novel xenogeneic leukemia model. Intravenous injection of EM-2eGFPluc into NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice resulted in significant dose-dependent bone marrow engraftment; lower levels engrafted in blood, lung, liver and spleen. In vitro-expanded human GDTc injected intraperitoneally were found at higher levels in blood and organs compared to those injected intravenously; GDTc survived at least 33 days post-injection. In therapy experiments, we documented decreased bone marrow leukemia burden in mice treated with GDTc. Live GDTc were found in spleen and bone marrow at endpoint, suggesting the potential usefulness of this therapy. PMID:21304898

  4. A Silica-Supported Iron Oxide Catalyst Capable of Activating Hydrogen Peroxide at Neutral pH Values

    PubMed Central

    Pham, Anh Le-Tuan; Lee, Changha; Doyle, Fiona M.; Sedlak, David L.

    2009-01-01

    Iron oxides catalyze the conversion of hydrogen peroxide (H2O2) into oxidants capable of transforming recalcitrant contaminants. Unfortunately, the process is relatively inefficient at circumneutral pH values due to competing reactions that decompose H2O2 without producing oxidants. Silica- and alumina-containing iron oxides prepared by sol-gel processing of aqueous solutions containing Fe(ClO4)3, AlCl3 and tetraethyl orthosilicate efficiently catalyzed the decomposition of H2O2 into oxidants capable of transforming phenol at circumneutral pH values. Relative to hematite, goethite and amorphous FeOOH, the silica-iron oxide catalyst exhibited a stoichiometric efficiency, defined as the number of moles of phenol transformed per mole of H2O2 consumed, that was 10 to 40 times higher than that of the iron oxides. The silica-alumina-iron oxide catalyst had a stoichiometric efficiency that was 50 to 80 times higher than that of the iron oxides. The significant enhancement in oxidant production is attributable to the interaction of Fe with Al and Si in the mixed oxides, which alters the surface redox processes, favoring the production of strong oxidants during H2O2 decomposition. PMID:19943668

  5. Radiopacity, pH and antimicrobial activity of Portland cement associated with micro- and nanoparticles of zirconium oxide and niobium oxide.

    PubMed

    Guerreiro Tanomaru, Juliane Maria; Storto, Inara; Da Silva, Guilherme Ferreira; Bosso, Roberta; Costa, Bernardo Cesar; Bernardi, Maria Inês Basso; Tanomaru-Filho, Mário

    2014-01-01

    The aim of this study was to evaluate some properties of the calcium silicate materials Mineral Trioxide Aggregate (MTA) and Portland cement (PC) with microparticulated (micro) and nanoparticulated (nano) zirconium oxide (ZrO2) or niobium oxide (Nb2O5). The experimental materials: White PC (PC), MTA-Angelus(®) (MTA), PC+ZrO2micro, PC+ZrO2nano, PC+Nb2O5micro and PC+Nb2O5nano were submitted to radiopacity and pH evaluations. Furthermore, the antimicrobial activity against different microorganisms was assessed by agar diffusion test. MTA presented higher radiopacity than other materials. However, all materials except PC presented higher radiopacity than recommended by ISO/ADA. MTA promoted higher pH values in all analyzed periods (p≤0.05). At the initial periods, PC and PC+ZrO2micro showed pH similar to MTA. All materials showed antimicrobial activity against the evaluated microorganisms. In conclusion, ZrO2 and Nb2O5 could be alternative radiopacifiers to be added to calcium silicate materials.

  6. A beta-galactoside alpha2,6-sialyltransferase produced by a marine bacterium, Photobacterium leiognathi JT-SHIZ-145, is active at pH 8.

    PubMed

    Yamamoto, Takeshi; Hamada, Yoko; Ichikawa, Masako; Kajiwara, Hitomi; Mine, Toshiki; Tsukamoto, Hiroshi; Takakura, Yoshimitsu

    2007-11-01

    A gene encoding a sialyltransferase produced by Photobacterium leiognathi JT-SHIZ-145 was cloned, sequenced, and expressed in Escherichia coli. The sialyltransferase gene contained an open reading frame of 1494 base pairs (bp) encoding a predicted protein of 497 amino acid residues. The deduced amino acid sequence of the sialyltransferase had no significant similarity to mammalian sialyltransferases and did not contain sialyl motifs, but did show high homology to another marine bacterial sialyltransferase, a beta-galactoside alpha2,6-sialyltransferase produced by P. damselae JT0160. The acceptor substrate specificity of the new enzyme was similar to that of the alpha2,6-sialyltransferase from P. damselae JT0160, but its activity was maximal at pH 8. This property is quite different from the properties of all mammalian and bacterial sialyltransferases reported previously, which have maximal activity at acidic pH. In general, both sialosides and cytidine-5'-monophospho-N-acetylneuraminic acid, the common donor substrate of sialyltransferases, are more stable under basic conditions. Therefore, a sialyltransferase with an optimum pH in the basic range should be useful for the preparation of sialosides and the modification of glycoconjugates, such as asialo-glycoproteins and asialo-glycolipids. Thus, the sialyltransferase obtained from P. leiognathi JT-SHIZ-145 is a promising tool for the efficient production of sialosides.

  7. Chemical equilibrium model for interfacial activity of crude oil in aqueous alkaline solution: the effects of pH, alkali and salt

    SciTech Connect

    Chan, M.; Yen, T.F.

    1980-11-01

    A chemical equilibrium model for interfacial activity of crude in aqueous alkaline solution is proposed. The model predicts the observed effects of pH and concentrations of alkali and salt on the interfacial tension (IFT). The model proposed was shown to describe the observed effects of acid content, pH, and sodium ions on the interfacial activity of crude oil in water. Once the pH of the interface reaches the pKa of the acids, sometimes with the help of addition of some salt, the IFT experiences a sudden steep drop to the range of 10/sup -2/ dynes/cm. After that, further addition of sodium either in the form of NaOH or NaCl is going to increase the IFT due to a shift of equilibriumn to the formation of undissociated soap. This was confirmed by the difference in the observed effect of sodium on the IFT of the extracted soap molecules which are dissociated easily and those which are associated highly and precipitated easily. These soap molecules have dissociation constant values ranging from below 10/sup -2/ to above one. 13 references.

  8. Unliganded and substrate bound structures of the cellooligosaccharide active lytic polysaccharide monooxygenase LsAA9A at low pH.

    PubMed

    Frandsen, Kristian E H; Poulsen, Jens-Christian N; Tandrup, Tobias; Lo Leggio, Leila

    2017-03-24

    Lytic polysaccharide monooxygenases (LPMOs) have been found to be key components in microbial (bacterial and fungal) degradation of biomass. They are copper metalloenzymes that degrade polysaccharides oxidatively and act in synergy with glycoside hydrolases. Recently crystallographic studies carried out at pH 5.5 of the LPMO from Lentinus similis belonging to the fungal LPMO family AA9 have provided the first atomic resolution view of substrate-LPMO interactions. The LsAA9A structure presented here determined at pH 3.5 shows significant disorder of the active site in the absence of substrate ligand. Furthermore some differences are also observed in regards to substrate (cellohexaose) binding, although the major interaction with the N-terminal histidine remains unchanged.

  9. Plasma membrane of Beta vulgaris storage root shows high water channel activity regulated by cytoplasmic pH and a dual range of calcium concentrations.

    PubMed

    Alleva, Karina; Niemietz, Christa M; Sutka, Moira; Maurel, Christophe; Parisi, Mario; Tyerman, Stephen D; Amodeo, Gabriela

    2006-01-01

    Plasma membrane vesicles isolated by two-phase partitioning from the storage root of Beta vulgaris show atypically high water permeability that is equivalent only to those reported for active aquaporins in tonoplast or animal red cells (Pf=542 microm s(-1)). The values were determined from the shrinking kinetics measured by stopped-flow light scattering. This high Pf was only partially inhibited by mercury (HgCl2) but showed low activation energy (Ea) consistent with water permeation through water channels. To study short-term regulation of water transport that could be the result of channel gating, the effects of pH, divalent cations, and protection against dephosphorylation were tested. The high Pf observed at pH 8.3 was dramatically reduced by medium acidification. Moreover, intra-vesicular acidification (corresponding to the cytoplasmic face of the membrane) shut down the aquaporins. De-phosphorylation was discounted as a regulatory mechanism in this preparation. On the other hand, among divalent cations, only calcium showed a clear effect on aquaporin activity, with two distinct ranges of sensitivity to free Ca2+ concentration (pCa 8 and pCa 4). Since the normal cytoplasmic free Ca2+ sits between these ranges it allows for the possibility of changes in Ca2+ to finely up- or down-regulate water channel activity. The calcium effect is predominantly on the cytoplasmic face, and inhibition corresponds to an increase in the activation energy for water transport. In conclusion, these findings establish both cytoplasmic pH and Ca2+ as important regulatory factors involved in aquaporin gating.

  10. Strategy for sensor based on fluorescence emission red shift of conjugated polymers: applications in pH response and enzyme activity detection.

    PubMed

    Tang, Yanli; Liu, Yue; Cao, Ali

    2013-01-15

    A new strategy was developed and applied in monitoring pH response and enzyme activity based on fluorescence emission red shift (FERS) of the conjugated polymer PPP-OR10 induced by the inner filter effect (IFE) of nitrobenzene derivatives. Neutral poly(p-phenylenes) functionalized with oligo(oxyethylene) side chains (PPP-OR10) was designed and synthesized by the Suzuki cross-coupling reaction. Nitrobenzene derivatives display different light absorption activities in the acidic or basic form due to adopting different electron-transition types. When environmental pH is higher than their pK(a) values, nitrobenzene derivatives exhibit strong absorbance around 400 nm, which is close to the maximal emission of polymer PPP-OR10. As a result, the maximal emission wavelength of PPP-OR10/nitrobenzene derivatives red shifts with the pH value increasing. Apparently, the IFE plays a very important role in this case. A new method has been designed that takes advantage of this pH-sensitive platform to sensor α-chymotrypsin (ChT) based on the IFE of p-nitroaniline, since the absorption spectrum of p-nitroaniline, the ChT-hydrolyzed product of N-benzoyl-L-tyrosine-p-nitroaniline (BTNA), overlaps with the emission spectrum of PPP-OR10. In addition, the present approach can detect α-chymotrypsin with a detection limit of 0.1 μM, which is lower than that of the corresponding absorption spectroscopy method. Furthermore, the pH response and enzyme detections can be carried out in 10% serum, which makes this new FERS-based strategy promising in applications in more complex conditions and a broader field.

  11. Growth of Pseudomonas chloritidismutans AW-1(T) on n-alkanes with chlorate as electron acceptor.

    PubMed

    Mehboob, Farrakh; Junca, Howard; Schraa, Gosse; Stams, Alfons J M

    2009-06-01

    Microbial (per)chlorate reduction is a unique process in which molecular oxygen is formed during the dismutation of chlorite. The oxygen thus formed may be used to degrade hydrocarbons by means of oxygenases under seemingly anoxic conditions. Up to now, no bacterium has been described that grows on aliphatic hydrocarbons with chlorate. Here, we report that Pseudomonas chloritidismutans AW-1(T) grows on n-alkanes (ranging from C7 until C12) with chlorate as electron acceptor. Strain AW-1(T) also grows on the intermediates of the presumed n-alkane degradation pathway. The specific growth rates on n-decane and chlorate and n-decane and oxygen were 0.5 +/- 0.1 and 0.4 +/- 0.02 day(-1), respectively. The key enzymes chlorate reductase and chlorite dismutase were assayed and found to be present. The oxygen-dependent alkane oxidation was demonstrated in whole-cell suspensions. The strain degrades n-alkanes with oxygen and chlorate but not with nitrate, thus suggesting that the strain employs oxygenase-dependent pathways for the breakdown of n-alkanes.

  12. Historical Allotment Gardens in Wrocław - The Need to Protection

    NASA Astrophysics Data System (ADS)

    Kononowicz, Wanda; Gryniewicz-Balińska, Katarzyna

    2016-06-01

    Since about the mid-nineteenth century, together with the changing socio-economic situation, different types of allotments appeared in Wrocław. Initially, they were rented gardens, gardens for the poor or for factory workers. At the beginning of the twentieth century, school gardens and the so called Schreber gardens with a large common square were set up as part of Dr. Schreber's educational health program. In 1914-1918, "war" vegetable gardens were commonly cultivated. In the 1920s allotment gardens began to be systematically introduced to the city plan as permanent, purposefully designed elements of urban greenery. They were often designed together with urban parks, or so called "Folk Parks" of a recreational and sport character. In the 1930s, during the economic crisis, allotments with garden houses were adapted for the unemployed and the homeless to live in. Wrocław allotment gardens have undeniable historical, social, recreational, economic and compositional value. These gardens are a cultural heritage that should be protected. In Western Europe we are witnessing a renaissance of the idea of allotments, while in Poland - a tendency to eliminate them from urban landscapes.

  13. Meaning and Measurability of Single-Ion Activities, the Thermodynamic Foundations of pH, and the Gibbs Free Energy for the Transfer of Ions between Dissimilar Materials

    PubMed Central

    Rockwood, Alan L

    2015-01-01

    Considering the relationship between concentration and vapor pressure (or the relationship between concentration and fugacity) single-ion activity coefficients are definable in purely thermodynamic terms. The measurement process involves measuring a contact potential between a solution and an external electrode. Contact potentials are measurable by using thermodynamically reversible processes. Extrapolation of an equation to zero concentration and ionic strength enables determination of single-ion activity coefficients. Single-ion activities can be defined and measured without using any extra-thermodynamic assumptions, concepts, or measurements. This method could serve as a gold standard for the validation of extra-thermodynamic methods for determining single-ion activities. Furthermore, it places the concept of pH on a thermodynamically solid foundation. Contact potential measurements can also be used to determine the Gibbs free energy for the transfer of ions between dissimilar materials. PMID:25919971

  14. cAMP/protein kinase A activates cystic fibrosis transmembrane conductance regulator for ATP release from rat skeletal muscle during low pH or contractions.

    PubMed

    Tu, Jie; Lu, Lin; Cai, Weisong; Ballard, Heather J

    2012-01-01

    We have shown that cystic fibrosis transmembrane conductance regulator (CFTR) is involved in ATP release from skeletal muscle at low pH. These experiments investigate the signal transduction mechanism linking pH depression to CFTR activation and ATP release, and evaluate whether CFTR is involved in ATP release from contracting muscle. Lactic acid treatment elevated interstitial ATP of buffer-perfused muscle and extracellular ATP of L6 myocytes: this ATP release was abolished by the non-specific CFTR inhibitor, glibenclamide, or the specific CFTR inhibitor, CFTR(inh)-172, suggesting that CFTR was involved, and by inhibition of lactic acid entry to cells, indicating that intracellular pH depression was required. Muscle contractions significantly elevated interstitial ATP, but CFTR(inh)-172 abolished the increase. The cAMP/PKA pathway was involved in the signal transduction pathway for CFTR-regulated ATP release from muscle: forskolin increased CFTR phosphorylation and stimulated ATP release from muscle or myocytes; lactic acid increased intracellular cAMP, pCREB and PKA activity, whereas IBMX enhanced ATP release from myocytes. Inhibition of PKA with KT5720 abolished lactic-acid- or contraction-induced ATP release from muscle. Inhibition of either the Na(+)/H(+)-exchanger (NHE) with amiloride or the Na(+)/Ca(2+)-exchanger (NCX) with SN6 or KB-R7943 abolished lactic-acid- or contraction-induced release of ATP from muscle, suggesting that these exchange proteins may be involved in the activation of CFTR. Our data suggest that CFTR-regulated release contributes to ATP release from contracting muscle in vivo, and that cAMP and PKA are involved in the activation of CFTR during muscle contractions or acidosis; NHE and NCX may be involved in the signal transduction pathway.

  15. Extracellular pH alkalinization by Cl-/HCO3- exchanger is crucial for TASK2 activation by hypotonic shock in proximal cell lines from mouse kidney.

    PubMed

    L'Hoste, S; Barriere, H; Belfodil, R; Rubera, I; Duranton, C; Tauc, M; Poujeol, C; Barhanin, J; Poujeol, P

    2007-02-01

    We have previously shown that K(+)-selective TASK2 channels and swelling-activated Cl(-) currents are involved in a regulatory volume decrease (RVD; Barriere H, Belfodil R, Rubera I, Tauc M, Lesage F, Poujeol C, Guy N, Barhanin J, Poujeol P. J Gen Physiol 122: 177-190, 2003; Belfodil R, Barriere H, Rubera I, Tauc M, Poujeol C, Bidet M, Poujeol P. Am J Physiol Renal Physiol 284: F812-F828, 2003). The aim of this study was to determine the mechanism responsible for the activation of TASK2 channels during RVD in proximal cell lines from mouse kidney. For this purpose, the patch-clamp whole-cell technique was used to test the effect of pH and the buffering capacity of external bath on Cl(-) and K(+) currents during hypotonic shock. In the presence of a high buffer concentration (30 mM HEPES), the cells did not undergo RVD and did not develop outward K(+) currents (TASK2). Interestingly, the hypotonic shock reduced the cytosolic pH (pH(i)) and increased the external pH (pH(e)) in wild-type but not in cftr (-/-) cells. The inhibitory effect of DIDS suggests that the acidification of pH(i) and the alkalinization of pH(e) induced by hypotonicity in wild-type cells could be due to an exit of HCO(3)(-). In conclusion, these results indicate that Cl(-) influx will be the driving force for HCO(3)(-) exit through the activation of the Cl(-)/HCO(3)(-) exchanger. This efflux of HCO(3)(-) then alkalinizes pH(e), which in turn activates TASK2 channels.

  16. Dependence on the F0F1-ATP synthase for the activities of the hydrogen-oxidizing hydrogenases 1 and 2 during glucose and glycerol fermentation at high and low pH in Escherichia coli.

    PubMed

    Trchounian, Karen; Pinske, Constanze; Sawers, R Gary; Trchounian, Armen

    2011-12-01

    Escherichia coli has four [NiFe]-hydrogenases (Hyd); three of these, Hyd-1, Hyd-2 and Hyd-3 have been characterized well. In this study the requirement for the F(0)F(1)-ATP synthase for the activities of the hydrogen-oxidizing hydrogenases Hyd-1 and Hyd-2 was examined. During fermentative growth on glucose at pH 7.5 an E. coli F(0)F(1)-ATP synthase mutant (DK8) lacked hydrogenase activity. At pH 5.5 hydrogenase activity was only 20% that of the wild type. Using in-gel activity staining, it could be demonstrated that both Hyd-1 and Hyd-2 were essentially inactive at these pHs, indicating that the residual activity at pH 5.5 was due to the hydrogen-evolving Hyd-3 enzyme. During fermentative growth in the presence of glycerol, hydrogenase activity in the mutant was highest at pH 7.5 attaining a value of 0.76 U/mg, or ~50% of wild type activity, and Hyd-2 was only partially active at this pH, while Hyd-1 was inactive. Essentially no hydrogenase activity was measured at pH 5.5 during growth with glycerol. At this pH the mutant had a hydrogenase activity that was maximally only ~10% of wild type activity with either carbon substrate but a weak activity of both Hyd-1 and Hyd-2 could be detected. Taken together, these results demonstrate for the first time that the activity of the hydrogen-oxidizing hydrogenases in E. coli depends on an active F(0)F(1)-ATP synthase during growth at high and low pH.

  17. Effect of pH and surface chemistry on the mechanism of H{sub 2}S removal by activated carbons

    SciTech Connect

    Adib, F.; Bagreev, A.; Bandosz, T.J.

    1999-08-15

    The performances of three wood-based activated carbons as adsorbents of hydrogen sulfide were evaluated by dynamic breakthrough testing. The subsequent products of H{sub 2}S oxidation on the carbon surfaces were analyzed. The adsorbents were studied using sorption of nitrogen, thermal analysis, Boehm titration, FTIR, ion chromatography, and temperature programmed desorption. Based on the results, the effects of surface chemistry and structural features on the yield of water soluble products and on the regenerability of the exhausted carbons were evaluated. The results showed that the breakthrough capacity and the yield on regeneration depend on the average pH of the carbon surface related to the pH in local pore environment. When the surface is very acidic, the dissociation of H{sub 2}S is suppressed resulting in a very small concentration of hydrogen sulfide ions and thus in the formation of highly dispersed sulfur. Such conditions are favorable for oxidation of sulfur to S{sup 4+} and S{sup 6+}. When the surface is less acidic the degree of dissociation is higher and the creation of polymeric elemental sulfur species resistant to further oxidation is more favorable. A small increase in pH (half a unit) in the acidic range results in a 15-fold increase in hydrogen sulfide breakthrough capacity accompanied by only a one third decrease in the yield of sulfur oxides.

  18. Electrochemical and spectroscopic investigations of isoniazide and its analogs with ds.DNA at physiological pH: evaluation of biological activities.

    PubMed

    Arshad, Nasima; Yunus, Uzma; Razzque, Shumaila; Khan, Maliha; Saleem, Samreen; Mirza, Bushra; Rashid, Naghmana

    2012-01-01

    Interaction and binding of isonicotinic acid hydrazide (INH) and its two analogs; pyrazine carboxylic acid hydrazide (PCH) and 2,4-dihydroxy benzoic acid hydrazide (2,4-DHBAH) with DNA has been investigated by UV-spectroscopy and cyclic voltammetry (CV) at physiological conditions of pH and temperature. Experimental results from both techniques were in good agreement and indicated stronger binding and formation of hydrazides-DNA complexes via intercalation. Among three hydrazides, 2,4-DHBAH showed greater interaction toward DNA at stomach pH (4.7) as evident from its comparatively greater binding constant, {K(b); 2.02 × 10(4) M(-1) (UV), 3.13 × 10(4) M(-1) (CV)}. The greater binding site size (n = 3) for 2,4-DHBAH at stomach pH inferred 3:1 binding stoichiometry and possibility of electrostatic interactions or hydrogen bonding along with intercalative mode of interaction between 2,4-DHBAH and DNA. The free energies of hydrazides-DNA complexes indicated the spontaneity of their binding. 2,4-DHBAH has shown promising anti-bacterial activities while anti-oxidant and cytotoxic potentials were exhibited by all three hydrazides.

  19. Role of H(+)-pyrophosphatase activity in the regulation of intracellular pH in a scuticociliate parasite of turbot: Physiological effects.

    PubMed

    Mallo, Natalia; Lamas, Jesús; de Felipe, Ana-Paula; Sueiro, Rosa-Ana; Fontenla, Francisco; Leiro, José-Manuel

    2016-10-01

    The scuticociliatosis is a very serious disease that affects the cultured turbot, and whose causal agent is the anphizoic and marine euryhaline ciliate Philasterides dicentrarchi. Several protozoans possess acidic organelles that contain high concentrations of pyrophosphate (PPi), Ca(2+) and other elements with essential roles in vesicular trafficking, pH homeostasis and osmoregulation. P. dicentrarchi possesses a pyrophosphatase (H(+)-PPase) that pumps H(+) through the membranes of vacuolar and alveolar sacs. These compartments share common features with the acidocalcisomes described in other parasitic protozoa (e.g. acid content and Ca(2+) storage). We evaluated the effects of Ca(2+) and ATP on H (+)-PPase activity in this ciliate and analyzed their role in maintaining intracellular pH homeostasis and osmoregulation, by the addition of PPi and inorganic molecules that affect osmolarity. Addition of PPi led to acidification of the intracellular compartments, while the addition of ATP, CaCl2 and bisphosphonates analogous of PPi and Ca(2+) metabolism regulators led to alkalinization and a decrease in H(+)-PPase expression in trophozoites. Addition of NaCl led to proton release, intracellular Ca(2+) accumulation and downregulation of H(+)-PPase expression. We conclude that the regulation of the acidification of intracellular compartments may be essential for maintaining the intracellular pH homeostasis necessary for survival of ciliates and their adaptation to salt stress, which they will presumably face during the endoparasitic phase, in which the salinity levels are lower than in their natural environment.

  20. Dependence of Ethanolic Fermentation, Cytoplasmic pH Regulation, and Viability on the Activity of Alcohol Dehydrogenase in Hypoxic Maize Root Tips 1

    PubMed Central

    Roberts, Justin K. M.; Chang, Keejong; Webster, Cecelia; Callis, Judy; Walbot, Virginia

    1989-01-01

    We examined the role of alcohol dehydrogenase (ADH) in the metabolism and survival of hypoxic maize (Zea mays L.) root tips. The dependence of the rate of ethanolic fermentation, cytoplasmic pH, and viability on the activity of ADH in maize root tips during extreme hypoxia was determined. Maize lines with ADH activities differing over about a 200-fold range were studied. Effects of genetic background were controlled by comparing pairs of F4 progeny of crosses between mutant (low ADH activity) and reference inbred lines. The capacity of hypoxic root tips to perform ethanolic fermentation exhibited a dependence on ADH activity only at activities found in Adh 1 nulls. The ability of maize root tips to withstand prolonged and extreme hypoxia was like-wise independent of ADH activity, except at the lowest activities. Root tips that exhibited lower tolerance of hypoxia had more acidic cytoplasm during extreme hypoxia. We conclude that the activity of ADH in normal maize root tips does not limit the capacity for energy production via fermentation, and does not determine viability under extreme hypoxia. The significance of the induction of ADH activity in plants by hypoxia is discussed. PMID:16666696

  1. Dependence of ethanolic fermentation, cytoplasmic pH regulation, and viability on the activity of alcohol dehydrogenase in hypoxic maize root tips

    SciTech Connect

    Roberts, J.; Chang, Keejong; Webster, C.; Callis, J.; Walbot, V. Stanford Univ., CA )

    1989-04-01

    We examined the role of alcohol dehydrogenase (ADH) in the metabolism and survival of hypoxic maize (Zea mays L.) root tips. The dependence of the rate of ethanolic fermentation, cytoplasmic pH, and viability on the activity of ADH in maize root tips during extreme hypoxia was determined. Maize lines with ADH activities differing over about a 200-fold range were studied. Effects of genetic background were controlled by comparing pairs of F4 progeny of crosses between mutant (low ADH activity) and reference inbred lines. The capacity of hypoxic root tips to perform ethanolic fermentation exhibited a dependence on ADH activity only at activities found in Adh 1 nulls. The ability of maize root tips to withstand prolonged and extreme hypoxia was likewise independent of ADH activity, except at the lowest activities. Root tips that exhibited lower tolerance of hypoxia had more acidic cytoplasm during extreme hypoxia. We conclude that the activity of ADH in normal maize root tips does not limit the capacity for energy production via fermentation, and does not determine viability under extreme hypoxia. The significance of the induction of ADH activity in plants by hypoxia is discussed.

  2. All-weld-metal design for AWS E10018M, E11018M and E12018M type electrodes

    SciTech Connect

    Surian, E.S.; Vedia, L.A. de

    1999-06-01

    This paper presents the results of a research program conducted to design the all-weld metal deposited with AWS A5.5-81 E10018M, E11018M and E12018M SMAW-type electrodes. The role that different alloying elements such as manganese, carbon and chromium play on the tensile properties, hardness and toughness as well as on the microstructure was studied. Criteria for selecting the weld metal composition leading to optimum combination of tensile strength and toughness are suggested. The effect of the variation of heat input, within the requirements of the AWS standard, on the mentioned properties was also analyzed. It was found that the E11018M and E12018M all-weld-metal tensile properties are very sensitive to variations in heat input. For certain values of chemical composition, welding parameter ranges suitable to guarantee the fulfillment of AWS requirements were determined.

  3. Quenched Assembly of NIR-Active Gold Nanoclusters Capped with Strongly Bound Ligands by Tuning Particle Charge via pH and Salinity.

    PubMed

    Stover, Robert J; Murthy, Avinash K; Nie, Golay D; Gourisankar, Sai; Dear, Barton J; Truskett, Thomas M; Sokolov, Konstantin V; Johnston, Keith P

    2014-07-03

    Gold nanospheres coated with a binary monolayer of bound citrate and cysteine ligands were assembled into nanoclusters, in which the size and near-infrared (NIR) extinction were tuned by varying the pH and concentration of added NaCl. During full evaporation of an aqueous dispersion of 4.5 ± 1.8 nm Au primary particles, the nanoclusters were formed and quenched by the triblock copolymer polylactic acid (PLA)(1K)-b-poly(ethylene glycol) (PEG)(10K)-b-PLA(1K), which also provided steric stabilization. The short-ranged depletion and van der Waals attractive forces were balanced against longer ranged electrostatic repulsion to tune the nanocluster diameter and NIR extinction. Upon lowering the pH from 7 to 5 at a given salinity, the magnitude of the charge on the primary particles decreased, such that the weaker electrostatic repulsion increased the hydrodynamic diameter and, consequently, NIR extinction of the clusters. At a given pH, as the concentration of NaCl was increased, the NIR extinction decreased monotonically. Furthermore, the greater screening of the charges on the nanoclusters weakened the interactions with PLA(1K)-b-PEG(10K)-b-PLA(1K) and thus lowered the amount of adsorbed polymer on the nanocluster surface. The generalization of the concept of self-assembly of small NIR-active nanoclusters to include a strongly bound thiol and the manipulation of the morphologies and NIR extinction by variation of pH and salinity not only is of fundamental interest but also is important for optical biomedical imaging and therapy.

  4. Isolated secretion granules from parotid glands of chronically stimulated rats possess an alkaline internal pH and inward-directed H/sup +/ pump activity

    SciTech Connect

    Arvan, P.; Castle, J.D.

    1986-10-01

    Secretion granules have been isolated from the parotid glands of rats that have been chronically stimulated with the ..beta..-adrenergic agonist, isoproterenol. These granules are of interest because they package a quantitatively different set of secretory proteins in comparison with granules from the normal gland. Polypeptides enriched in proline, glycine, and glutamine, which are known to have pI's >10, replace ..cap alpha..-amylase (pI's = 6.8) as the principal content species. The internal pH of granules from the treated rats changes from 7.8 in a potassium sulfate medium to 6.9 in a choline chloride medium. The increased pH over that of normal parotid granules (approx.6.8) appears to protect the change in composition of the secretory contents. Whereas normal mature parotide granules have practically negligible levels of H/sup +/ pumping ATPase activity, the isolated granules from isoproterenol-treated rats undergo a time-dependent internal acidification that requires the presence of ATP and is abolished by an H/sup +/ ionophore. Additionally, an inside-positive granule transmembrane potential develops after ATP addition that depends upon ATP hydrolysis. Two independent methods have been used that exclude the possibility that contaminating organelles are the source of the H/sup +/-ATPase activity. Together these data provide clear evidence for the presence of an H/sup +/ pump in the membranes of parotid granules from chronically stimulated rats. However, despite the presence of H/sup +/-pump activity, fluorescence microscopy with the weak base, acridine orange, reveals that the intragranular pH in live cells is greater than that of the cytoplasm.

  5. Endophytic fungi producing of esterases: evaluation in vitro of the enzymatic activity using pH indicator.

    PubMed

    Lisboa, Helen Cristina Fávero; Biasetto, Carolina Rabal; de Medeiros, João Batista; Âraújo, Angela Regina; Silva, Dulce Helena Siqueira; Teles, Helder Lopes; Trevisan, Henrique Celso

    2013-01-01

    A sensitive and efficient colorimetric method was optimized for detection of esterase enzymes produced by endophytic fungi for development of High-Throughput Screening (HTS). The fungi were isolated and obtained previously from plant species of Cerrado and Atlantic Forest located in areas of environmental preservation in the State of Sao Paulo / Brazil, as part of the project "Chemical and biological prospecting endophytic fungi associated to plant species of Cerrado and Atlantic Forest". The compounds ethyl butyrate, ethyl acetate and methyl propionate were used as standards esters which were hydrolyzed by extracellular enzyme from endophytic fungi (EC. 3.1.1.1--carboxyl-esterases) for production of carboxylic acids. Thus, the reduction of the pH increases the protonated indicator concentration (bromothymol blue), changing the color of the reaction medium (from blue to yellow), that can be observed and measured by spectrophotometry at 616 nm. The methodology with acid-base indicator was performed on 13 microorganisms, aiming Periconia atropurpurea as a potential source of esterase for biotransformation of short chain esters. The results also evidenced that this methodology showed to be efficient, fast, cheap, having low consumption of reagents and easy development, and can be applied to screen carboxylic-ester hydrolases in a large number of microorganisms.

  6. Towards a metagenomic understanding on enhanced biomethane production from waste activated sludge after pH 10 pretreatment

    PubMed Central

    2013-01-01

    Background Understanding the effects of pretreatment on anaerobic digestion of sludge waste from wastewater treatment plants is becoming increasingly important, as impetus moves towards the utilization of sludge for renewable energy production. Although the field of sludge pretreatment has progressed significantly over the past decade, critical questions concerning the underlying microbial interactions remain unanswered. In this study, a metagenomic approach was adopted to investigate the microbial composition and gene content contributing to enhanced biogas production from sludge subjected to a novel pretreatment method (maintaining pH at 10 for 8 days) compared to other documented methods (ultrasonic, thermal and thermal-alkaline). Results Our results showed that pretreated sludge attained a maximum methane yield approximately 4-fold higher than that of the blank un-pretreated sludge set-up at day 17. Both the microbial and metabolic consortium shifted extensively towards enhanced biodegradation subsequent to pretreatment, providing insight for the enhanced methane yield. The prevalence of Methanosaeta thermophila and Methanothermobacter thermautotrophicus, together with the functional affiliation of enzymes-encoding genes suggested an acetoclastic and hydrogenotrophic methanogenesis pathway. Additionally, an alternative enzymology in Methanosaeta was observed. Conclusions This study is the first to provide a microbiological understanding of improved biogas production subsequent to a novel waste sludge pretreatment method. The knowledge garnered will assist the design of more efficient pretreatment methods for biogas production in the future. PMID:23506434

  7. Inhibition of potato polyphenol oxidase by anions and activity in various carboxylate buffers (pH 4.8) at constant ionic strength.

    PubMed

    Malkin, B D; Thickman, K R; Markworth, C J; Wilcox, D E; Kull, F J

    2001-01-01

    The activity of potato polyphenol oxidase (tyrosinase) toward DL-3,4-dihydroxyphenylalanine (K(M) 5.39 mM) was studied using a variety of carboxylate buffers at a common pH and ionic strength. Enzyme activity, greatest in citrate and least in oxalate, correlated with increasing carboxyl concentration and molecular mass. The lower activity in oxalate was attributed to more effective chelation of a copper(II) form of the enzyme by the oxalate dianion. Sodium halide salts inhibited the enzyme. Although there was little difference in inhibition between sodium and potassium salts, the degree and type of inhibition was anion dependent; K(is), values for NaCl and KCl, (competitive inhibitors) were 1.82 and 1.62 mM, whereas Na(2) SO(4) and K(2) SO(4) (mixed inhibitors) had K(is) and K(ii) values in the 250 to 450 mM range.

  8. Ultrasonic Examination of Double-Shell Tank 241-AW-103. Examination Completed September 2006

    SciTech Connect

    Pardini, Allan F.; Posakony, Gerald J.; Weier, Dennis R.

    2007-04-01

    AREVA NC Inc. (AREVA), under a contract from CH2M Hill Hanford Group (CH2M Hill), has performed an ultrasonic examination of selected portions of Double-Shell Tank 241-AW-103. PNNL is responsible for preparing a report(s) that describes the results of the AREVA ultrasonic examinations. The purpose of this examination was to provide information that could be used to evaluate the integrity of the wall of the primary and secondary tank. The requirements for the ultrasonic examination of Tank 241-AW-103 were to detect, characterize (identify, size, and locate), and record measurements made of any wall thinning, pitting, or cracks that might be present in the wall of the primary tank and the wall of the secondary tank. Any measurements that exceed the requirements set forth in the Engineering Task Plan (ETP), RPP-Plan-27202 (Jensen 2005) and summarized on page 1 of this document, are to be reported to CH2M Hill and the Pacific Northwest National Laboratory (PNNL) for further evaluation. Under the contract with CH2M Hill, all data is to be recorded on electronic media and paper copies of all measurements are provided to PNNL for third-party evaluation. PNNL is responsible for preparing a report(s) that describes the results of the AREVA ultrasonic examinations. The results of the examination of Tank 241-AW-103 have been evaluated by PNNL personnel. The primary tank ultrasonic examination consisted of two vertical 15-in.-wide scan paths over the entire height of the tank, the heat-affected zone (HAZ) of four vertical welds and one horizontal weld from Riser 29 and two vertical 15-in.-wide scan paths over the entire height of the tank from Riser 28. Additionally, two vertical 15-in.-wide scan paths over the entire height of the secondary tank from Riser 28 were performed. The examinations were performed to detect any wall thinning, pitting, or cracking in the primary tank wall.

  9. Ultrasonic Examination of Double-Shell Tank 241-AW-103 Examination Completed September 2006

    SciTech Connect

    Pardini, Allan F.; Posakony, Gerald J.

    2006-11-06

    AREVA NC Inc. (AREVA), under a contract from CH2M Hill Hanford Group (CH2M Hill), has performed an ultrasonic examination of selected portions of Double-Shell Tank 241-AW-103. PNNL is responsible for preparing a report(s) that describes the results of the AREVA ultrasonic examinations. The purpose of this examination was to provide information that could be used to evaluate the integrity of the wall of the primary and secondary tank. The requirements for the ultrasonic examination of Tank 241-AW-103 were to detect, characterize (identify, size, and locate), and record measurements made of any wall thinning, pitting, or cracks that might be present in the wall of the primary tank and the wall of the secondary tank. Any measurements that exceed the requirements set forth in the Engineering Task Plan (ETP), RPP-Plan-27202 (Jensen 2005) and summarized on page 1 of this document, are to be reported to CH2M Hill and the Pacific Northwest National Laboratory (PNNL) for further evaluation. Under the contract with CH2M Hill, all data is to be recorded on electronic media and paper copies of all measurements are provided to PNNL for third-party evaluation. PNNL is responsible for preparing a report(s) that describes the results of the AREVA ultrasonic examinations. The results of the examination of Tank 241-AW-103 have been evaluated by PNNL personnel. The primary tank ultrasonic examination consisted of two vertical 15-in.-wide scan paths over the entire height of the tank, the heat-affected zone (HAZ) of four vertical welds and one horizontal weld from Riser 29 and two vertical 15-in.-wide scan paths over the entire height of the tank from Riser 28. Additionally, two vertical 15-in.-wide scan paths over the entire height of the secondary tank from Riser 28 were performed. The examinations were performed to detect any wall thinning, pitting, or cracking in the primary tank wall.

  10. Intracellular pH regulation by HCO3-/Cl- exchange is activated during early mouse zygote development.

    PubMed

    Phillips, K P; Baltz, J M

    1999-04-15

    We report here that at least one major pHi-regulatory mechanism, the HCO3-/Cl- exchanger, is quiescent in unfertilized mouse eggs but becomes fully activated during early development following fertilization. Zygotes (8-12 h postfertilization) exhibited a marked intracellular alkalinization upon external Cl- removal, which is indicative of active HCO3-/Cl- exchangers, in contrast to the very small response observed in eggs. In addition, efflux of Cl- from eggs upon external Cl- removal was much slower than that from zygotes, indicating additional pathways for Cl- to cross the plasma membrane in zygotes. Furthermore, while zygotes quickly recovered from an induced alkalosis, eggs exhibited only a slow, incomplete recovery. Following in vitro fertilization (IVF), increased HCO3-/Cl- exchanger activity was first detectable about 4 h postfertilization and reached the maximal level after about 8 h. The upregulation of HCO3-/Cl- exchanger activity after fertilization appeared to occur by activation of existing, inactive exchangers rather than by synthesis or transport of new exchangers, as the increase in activity following IVF was unaffected by inhibition of protein synthesis or by disruption of the Golgi apparatus or the cytoskeleton. This activation may depend on the Ca2+ transients which follow fertilization, as suppression of these transients, using the Ca2+ chelator BAPTA, reduced subsequent upregulation of HCO3-/Cl- exchanger activity by about 50%. Activation of pHi-regulatory systems may be a widespread feature of the earliest period of embryonic development, not restricted to species such as marine invertebrates as previously believed.

  11. Dissociation energies of PH and PH+.

    NASA Astrophysics Data System (ADS)

    Reddy, R. R.; Nazeer Ahammed, Y.; Srinivasa Rao, A.; Rao, T. V. R.

    1995-12-01

    Dissociation energies for the ground electronic states of diatomic PH and PH+ are determined by fitting empirical potential functions to the respective RKRV curves using correlation coefficients. The estimated ground state dissociation energies of PH and PH+ are 3.10 and 3.20 eV respectively by the curve fitting procedure using the Lippincott potential function. The computed values are in good agreement with experimental values.

  12. The Role of Auxin, pH, and Stress in the Activation of Embryogenic Cell Division in Leaf Protoplast-Derived Cells of Alfalfa1

    PubMed Central

    Pasternak, Taras P.; Prinsen, Els; Ayaydin, Ferhan; Miskolczi, Pál; Potters, Geert; Asard, Han; Van Onckelen, Harry A.; Dudits, Dénes; Fehér, Attila

    2002-01-01

    Culturing leaf protoplast-derived cells of the embryogenic alfalfa (Medicago sativa subsp. varia A2) genotype in the presence of low (1 μm) or high (10 μm) 2, 4-dichlorophenoxyacetic acid (2,4-D) concentrations results in different cell types. Cells exposed to high 2,4-D concentration remain small with dense cytoplasm and can develop into proembryogenic cell clusters, whereas protoplasts cultured at low auxin concentration elongate and subsequently die or form undifferentiated cell colonies. Fe stress applied at nonlethal concentrations (1 mm) in the presence of 1 μm 2,4-D also resulted in the development of the embryogenic cell type. Although cytoplasmic alkalinization was detected during cell activation of both types, embryogenic cells could be characterized by earlier cell division, a more alkalic vacuolar pH, and nonfunctional chloroplasts as compared with the elongated, nonembryogenic cells. Buffering of the 10 μm 2,4-D-containing culture medium by 10 mm 2-(N-morpholino)ethanesulfonic acid delayed cell division and resulted in nonembryogenic cell-type formation. The level of endogenous indoleacetic acid (IAA) increased transiently in all protoplast cultures during the first 4 to 5 d, but an earlier peak of IAA accumulation correlated with the earlier activation of the division cycle in embryogenic-type cells. However, this IAA peak could also be delayed by buffering of the medium pH by 2-(N-morpholino)ethanesulfonic acid. Based on the above data, we propose the involvement of stress responses, endogenous auxin synthesis, and the establishment of cellular pH gradients in the formation of the embryogenic cell type. PMID:12177494

  13. Expression of human papilloma virus type 16 E5 protein in amelanotic melanoma cells regulates endo-cellular pH and restores tyrosinase activity

    PubMed Central

    Di Domenico, Fabio; Foppoli, Cesira; Blarzino, Carla; Perluigi, Marzia; Paolini, Francesca; Morici, Salvatrice; Coccia, Raffaella; Cini, Chiara; De Marco, Federico

    2009-01-01

    Background Melanin synthesis, the elective trait of melanocytes, is regulated by tyrosinase activity. In tyrosinase-positive amelanotic melanomas this rate limiting enzyme is inactive because of acidic endo-melanosomal pH. The E5 oncogene of the Human Papillomavirus Type 16 is a small transmembrane protein with a weak transforming activity and a role during the early steps of viral infections. E5 has been shown to interact with 16 kDa subunit C of the trans-membrane Vacuolar ATPase proton pump ultimately resulting in its functional suppressions. However, the cellular effects of such an interaction are still under debate. With this work we intended to explore whether the HPV16 E5 oncoprotein does indeed interact with the vacuolar ATPase proton pump once expressed in intact human cells and whether this interaction has functional consequences on cell metabolism and phenotype. Methods The expression of the HPV16-E5 oncoproteins was induced in two Tyrosinase-positive amelanotic melanomas (the cell lines FRM and M14) by a retroviral expression construct. Modulation of the intracellular pH was measured with Acridine orange and fluorescence microscopy. Expression of tyrosinase and its activity was followed by RT-PCR, Western Blot and enzyme assay. The anchorage-independence growth and the metabolic activity of E5 expressing cells were also monitored. Results We provide evidence that in the E5 expressing cells interaction between E5 and V-ATPase determines an increase of endo-cellular pH. The cellular alkalinisation in turn leads to the post-translational activation of tyrosinase, melanin synthesis and phenotype modulation. These effects are associated with an increased activation of tyrosine analogue anti-blastic drugs. Conclusion Once expressed within intact human cells the HPV16-E5 oncoprotein does actually interact with the vacuolar V-ATPase proton pump and this interaction induces a number of functional effects. In amelanotic melanomas these effects can modulate the

  14. Active microorganisms in forest soils differ from the total community yet are shaped by the same environmental factors: the influence of pH and soil moisture.

    PubMed

    Romanowicz, Karl J; Freedman, Zachary B; Upchurch, Rima A; Argiroff, William A; Zak, Donald R

    2016-10-01

    Predicting the impact of environmental change on soil microbial functions requires an understanding of how environmental factors shape microbial composition. Here, we investigated the influence of environmental factors on bacterial and fungal communities across an expanse of northern hardwood forest in Michigan, USA, which spans a 500-km regional climate gradient. We quantified soil microbial community composition using high-throughput DNA sequencing on coextracted rDNA (i.e. total community) and rRNA (i.e. active community). Within both bacteria and fungi, total and active communities were compositionally distinct from one another across the regional gradient (bacteria P = 0.01; fungi P < 0.01). Taxonomically, the active community was a subset of the total community. Compositional differences between total and active communities reflected changes in the relative abundance of dominant taxa. The composition of both the total and active microbial communities varied by site across the gradient (P < 0.01) and was shaped by differences in soil moisture, pH, SOM carboxyl content, as well as C and N concentration. Our study highlights the importance of distinguishing between metabolically active microorganisms and the total community, and emphasizes that the same environmental factors shape the total and active communities of bacteria and fungi in this ecosystem.

  15. Influence of rotary swaging and subsequent age hardening on properties of EN AW 6082 aluminium alloy

    NASA Astrophysics Data System (ADS)

    Maleček, L.; Palán, J.; Nacházel, J.; Dlouhý, J.

    2017-02-01

    Mechanical properties and microstructure of EN AW 6082 were investigated. The aluminium alloy was processed by combining the solution annealing, plastic deformation and artificial age hardening, respectively. The initial state of the investigated material was provided in the form of extruded rods with the diameter of 12 mm. For the solution annealing the temperature 530 °C was chosen. The plastic deformation was realized by rotary swaging at ambient temperature and the investigated material was rotary swaged from 12 to 10 mm in diameter. The effect of the age hardening temperature and time was studied at temperatures of 120 and 160 °C and times 1 - 12 hours. The impact of processing parameters on mechanical properties was assessed by tensile testing and hardness measurement. Metallographic examination was carried out by light optical microscopy (LOM) and scanning electron microscopy using electron backscatter diffraction (SEM-EBSD).

  16. Evaluation of medical image watermarking with tamper detection and recovery (AW-TDR).

    PubMed

    Zain, Jasni M; Fauzi, Abdul R M

    2007-01-01

    This paper will study and evaluate watermarking technique by Zain and Fauzi [1]. Recommendations will then be made to enhance the technique especially in the aspect of recovery or reconstruction rate for medical images. A proposal will also be made for a better distribution of watermark to minimize the distortion of the Region of Interest (ROI). The final proposal will enhance AW-TDR in three aspects; firstly the image quality in the ROI will be improved as the maximum change is only 2 bits in every 4 pixels, or embedding rate of 0.5 bits/pixel. Secondly the recovery rate will also be better since the recovery bits are located outside the region of interest. The disadvantage in this is that, only manipulation done in the ROI will be detected. Thirdly the quality of the reconstructed image will be enhanced since the average of 2 x 2 pixels would be used to reconstruct the tampered image.

  17. Prediction of traffic convective instability with spectral analysis of the Aw-Rascle-Zhang model

    NASA Astrophysics Data System (ADS)

    Belletti, Francois; Huo, Mandy; Litrico, Xavier; Bayen, Alexandre M.

    2015-10-01

    This article starts from the classical Aw-Rascle-Zhang (ARZ) model for freeway traffic and develops a spectral analysis of its linearized version. A counterpart to the Froude number in hydrodynamics is defined that enables a classification of the nature of vehicle traffic flow using the explicit solution resulting from the analysis. We prove that our linearization about an equilibrium is stable for congested regimes and unstable otherwise. NGSIM data for congested traffic trajectories is used so as to confront the linearized model's predictions to actual macroscopic behavior of traffic. The model is shown to achieve good accuracy for speed and flow. In particular, it accounts for the advection of oscillations on boundaries into the interior domain where the PDE under study is solved.

  18. [Definition of the history of medicine in the interpretation of Władysław Szumowski].

    PubMed

    Gryglewski, Ryszard W

    2009-01-01

    Władysław Szumowski is said to be one of the most eminent Polish historians of medicine. The present text is an attempt of describing Szumowski's ideas of the essence of the history of medicine, its place among sciences and a role it should play in education of future doctors. The paper presents both the definition and the methodology, which in Szumowski's opinion should result from the philosophy, and particularly from the logic. Szumowski is referring to these research inspirations, scientists and conceptions that had a significant influence on him. At the same time the author of the article is willing to extract an original thought of Szumowski and divide it from adopted ideas of other people's conceptions.

  19. Phosphatase activities of rat intestinal enterocytes and their relation to diverse luminal pH, with special references to the possible localization of phytase along the brush border membrane.

    PubMed

    Nakano, Y; Kawamoto, T; Takano, Y

    2001-12-01

    Phosphatase activities associated with the intestinal brush border membrane (BBM) of the rat were examined histochemically in relation to the characteristic environment of the intestine, where luminal pH fluctuates drastically between alkaline and acid pH ranges. Special attention was given to intestinal alkaline phosphatase (IALP) and phytase on the BBM. Whole body fresh-frozen sections of young rats and their rapidly frozen and freeze-substituted small intestines, embedded in Technovit 7100, were processed for the histochemical demonstration of phosphatase activity at three different pH values (9.2, 7.3, and 5.2), representing the deviation of luminal pH in vivo. Either an azo-dye method or lead-salt method was employed using naphthol AS-MX phosphate and ATP as substrate, respectively. With the azo-dye method, intense phosphatase reactions were demonstrated along the BBM at all three pH ranges. Phosphatase reactions of the BBM at pH 9.2 and 7.3 were abolished by L(+)-phenylalanine, heat pre-treatment, and EDTA chelation although some reaction remained at pH 7.3 after the treatment with EDTA or L(+)-phenylalanine. Phosphatase reactions of the BBM at pH 5.2 were resistant to L(+)-phenylalanine, L(+)-tartrate, PCMB and EDTA chelation, implying that the characteristics of the enzyme responsible for phosphohydrolysis at acid pH values differed from those at higher pH values. The lead-salt method in which ATP was used as substrate revealed intense reactions--which were dependent on Mg++ and stimulated by Ca++ and resistant to L(+)phenylalanine--to be localized along the BBM at alkaline and neutral pH values, but not at acid pH values. In vitro experiments showed progressive hydrolysis of naphthol AS-MX phosphate by purified phytase at pH 5.2, in a dose-dependent manner, and suggested the possible involvement of phytase in the phosphatase reactions of the BBM at acid pH. These data indicate that the phosphatase reactions at alkaline and neutral pH values, associated

  20. Purification and characterisation of a malto-oligosaccharide-forming amylase active at high pH from Bacillus clausii BT-21.

    PubMed

    Duedahl-Olesen, L; Kragh, K M; Zimmermann, W

    2000-10-20

    Bacillus clausii BT-21 produced an extracellular malto-oligosaccharide-forming amylase active at high pH when grown on starch substrates. The enzyme was purified to homogeneity by affinity and anion-exchange chromatography. The molecular weight of the enzyme estimated by sodium dodecyl sulfate polyacrylamide electrophoresis was 101 kDa. The enzyme showed an optimum of activity at pH 9.5 and 55 degrees C. Maltohexaose was detected as the main initially formed starch hydrolysis product. Maltotetraose and maltose were the main products obtained after hydrolysis of starch by the enzyme for an extended period of time and were not further degraded. The enzyme readily hydrolysed soluble starch, amylopectin and amylose, while cyclodextrins, pullulan or dextran were not degraded. The mode of action during hydrolysis of starch indicated an exo-acting type of amylolytic enzyme mainly producing maltohexaose and maltotetraose. Amino acid sequencing of the enzyme revealed high homology with the maltohexaose-forming amylase from Bacillus sp. H-167.

  1. Interconnected Co-Entrapped, N-Doped Carbon Nanotube Film as Active Hydrogen Evolution Cathode over the Whole pH Range.

    PubMed

    Xing, Zhicai; Liu, Qian; Xing, Wei; Asiri, Abdullah M; Sun, Xuping

    2015-06-08

    The use of electrocatalysts with low metal content (metal-deficient) or metal free for the hydrogen evolution reaction (HER) can prevent or decrease metal ion release, which reduces environmental impact; development of such catalysts with high activity and durability over the whole pH range is thus highly desired but still remains a huge challenge. Herein, we describe the direct growth of a film consisting of interconnected Co-entrapped, N-doped carbon nanotubes on carbon cloth using chemical vapor deposition from dicyanodiamine using a Co3 O4 nanowire array as catalyst. This integrated architecture is used as a flexible 3D electrode for the electrolytic hydrogen evolution with outstanding catalytic activity and durability in acidic media. Moreover, this electrode is also highly efficient under neutral and basic conditions. It offers us an attractive carbon-based metal-deficient HER catalyst outperforming most transition-metal and all metal-free/deficient catalysts.

  2. Charge screening by internal pH and polyvalent cations as a mechanism for activation, inhibition, and rundown of TRPM7/MIC channels.

    PubMed

    Kozak, J Ashot; Matsushita, Masayuki; Nairn, Angus C; Cahalan, Michael D

    2005-11-01

    The Mg2+-inhibited cation (MIC) current, believed to represent activity of TRPM7 channels, is found in lymphocytes and mast cells, cardiac and smooth muscle, and several other eukaryotic cell types. MIC current is activated during whole-cell dialysis with divalent-free internal solutions. Millimolar concentrations of intracellular Mg2+ (or other divalent metal cations) inhibit the channels in a voltage-independent manner. The nature of divalent inhibition and the mechanism of channel activation in an intact cell remain unknown. We show that the polyamines (spermine, spermidine, and putrescine) inhibit the MIC current, also in a voltage-independent manner, with a potency that parallels the number of charges. Neomycin and poly-lysine also potently inhibited MIC current in the absence of Mg2+. These same positively charged ions inhibited IRK1 current in parallel with MIC current, suggesting that they probably act by screening the head group phosphates on PIP2 and other membrane phospholipids. In agreement with this hypothesis, internal protons also inhibited MIC current. By contrast, tetramethylammonium, tetraethylammonium, and hexamethonium produced voltage-dependent block but no inhibition. We show that inhibition by internal polyvalent cations can be relieved by alkalinizing the cytosol using externally applied ammonium or by increasing pH in inside-out patches. Furthermore, in perforated-patch and cell-attached recordings, when intracellular Mg2+ is not depleted, endogenous MIC or recombinant TRPM7 currents are activated by cytosolic alkalinization and inhibited by acidification; and they can be reactivated by PIP2 following rundown in inside-out patches. We propose that MIC (TRPM7) channels are regulated by a charge screening mechanism and may function as sensors of intracellular pH.

  3. Tuning the Laplaza-Cummins 3-coordinate M[N(R)Ph]3 catalyst to activate and cleave CO2.

    PubMed

    Brookes, Nigel J; Ariafard, Alireza; Stranger, Robert; Yates, Brian F

    2011-05-28

    The Laplaza-Cummins catalyst L(3)Mo (L = N(R)Ar), is experimentally inactive towards carbon dioxide. Previous theoretical analysis identified the cause for this inactivity and suggested that a switch to a d(2) transition metal may induce activity towards the inert CO(2) molecule. In this manuscript we have tested this hypothesis by altering the central metal to Ta, Nb or V. Our calculations suggest that the tantalum analogue, TaL(3), will successfully bind to CO(2) in a mononuclear η(2) arrangement and, importantly, will strongly activate one C-O bond to a point where spontaneous C-O cleavage occurs. This prediction of a strongly exothermic reaction takes into consideration the initial barrier to formation, spin crossings, ligand bulk and even the choice of density functional in the calculations. The Nb analogue will likely coordinate CO(2) but reaction may not proceed further. In contrast, the V analogue faces an initial coordination barrier and is not expected to be sufficiently active to coordinate CO(2) to the triamide catalyst. A similar scenario exists for mixed metal interactions involving a d(2) and d(4) combination in a bridging dinuclear arrangement.

  4. 47 CFR 27.1176 - Cost-sharing requirements for AWS in the 2150-2160/62 MHz band.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... clearance of BRS incumbents from this spectrum, including BRS incumbents occupying the 2150-2162 MHz band on... relocation until the end of the period specified in § 27.1190. If an AWS licensee enters into a spectrum leasing arrangement and the spectrum lessee triggers a cost-sharing obligation, the licensee is the...

  5. 75 FR 3615 - Airworthiness Directives; Agusta S.p.A. (Agusta) Model AB139 and AW139 Helicopters

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-22

    ...; AD 2009-19-51] RIN 2120-AA64 Airworthiness Directives; Agusta S.p.A. (Agusta) Model AB139 and AW139... adopting Airworthiness Directive (AD) 2009-19-51, which was sent previously to all known U.S. owners and... the tail panels for debonding and, if the debonding area exceeds a certain limit, repairing...

  6. Replication mechanism and sequence analysis of the replicon of pAW63, a conjugative plasmid from Bacillus thuringiensis.

    PubMed

    Wilcks, A; Smidt, L; Okstad, O A; Kolsto, A B; Mahillon, J; Andrup, L

    1999-05-01

    A 5.8-kb fragment of the large conjugative plasmid pAW63 from Bacillus thuringiensis subsp. kurstaki HD73 containing all the information for autonomous replication was cloned and sequenced. By deletion analysis, the pAW63 replicon was reduced to a 4.1-kb fragment harboring four open reading frames (ORFs). Rep63A (513 amino acids [aa]), encoded by the largest ORF, displayed strong similarity (40% identity) to the replication proteins from plasmids pAMbeta1, pIP501, and pSM19035, indicating that the pAW63 replicon belongs to the pAMbeta1 family of gram-positive theta-replicating plasmids. This was confirmed by the facts that no single-stranded DNA replication intermediates could be detected and that replication was found to be dependent on host-gene-encoded DNA polymerase I. An 85-bp region downstream of Rep63A was also shown to have strong similarity to the origins of replication of pAMbeta1 and pIP501, and it is suggested that this region contains the bona fide pAW63 ori. The protein encoded by the second large ORF, Rep63B (308 aa), was shown to display similarity to RepB (34% identity over 281 aa) and PrgP (32% identity over 310 aa), involved in copy control of the Enterococcus faecalis plasmids pAD1 and pCF10, respectively. No significant similarity to known proteins or DNA sequences could be detected for the two smallest ORFs. However, the location, size, hydrophilicity, and orientation of ORF6 (107 codons) were analogous to those features of the putative genes repC and prgO, which encode stability functions on plasmids pAD1 and pCF10, respectively. The cloned replicon of plasmid pAW63 was stably maintained in Bacillus subtilis and B. thuringiensis and displayed incompatibility with the native pAW63. Hybridization experiments using the cloned replicon as a probe showed that pAW63 has similarity to large plasmids from other B. thuringiensis subsp. kurstaki strains and to a strain of B. thuringiensis subsp. alesti.

  7. EFFECTS OF pH AND OF VARIOUS CONCENTRATIONS OF SODIUM, POTASSIUM, AND CALCIUM CHLORIDE ON MUSCULAR ACTIVITY OF THE ISOLATED CROP OF PERIPLANETA AMERICANA (ORTHOPTERA)

    PubMed Central

    Griffiths, James T.; Tauber, Oscar E.

    1943-01-01

    1. Twenty-five solutions which contained KCl (0.0, 0.2, 0.4, 0.6, and 0.8 gm. per liter), in combination with CaCl2 (0.0, 0.2, 0.4, 0.6, and 0.8 gm. per liter), 10.0 gm. of NaCl, and 0.2 gm. of NaHCO3 per liter of solution were tested in order to determine satisfactory KCl/CaCl2 ratios in an insect physiological salt mixture for the maintenance of muscular activity by the isolated crop of the American roach. Satisfactory activity products (0.390 to 0.549) were obtained in seven mixtures with KCl/CaCl2 ratios of 0.2/0.2, 0.4/0.4, 0.6/0.6, 0.8/0.8, 0.2/0.4, 0.4/0.6, and 0.6/0.8, expressed as gram per liter. These ratios lie between 0.50 and 1.00. In solutions which contained calcium, but no potassium, approximately 50 per cent of the crops exhibited an initial tone increase and were arrested in rigor. See Fig. 2. In solutions which contained potassium, but no calcium, all crops showed an initial loss of tone and arrest in relaxation. See Fig. 2. 2. Seven KCl/CaCl2 ratios (see paragraph 1 above) were tested with eight NaCl concentrations (1.0, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, and 1.8 per cent) at a pH of 8.0. In these mixtures, the ones with KCl/CaCl2 ratios of less than 1.0 produced higher activity products than those with ratios equal to 1.00. The highest average activity product (0.849) was obtained in the solutions with 0.2 gm. of KCl and 0.4 gm. of CaCl2 per liter. 3. Four KCl/CaCl2 ratios (0.2/0.2, 0.4/0.4, 0.2/0.4, and 0.4/0.6 gm. per liter) were tested with 1.4, 1.5, and 1.6 per cent NaCl at a pH of 7.5. When analyzed with data from comparable solutions at a pH of 8.0, it was found that 1.4 per cent NaCl afforded an optimum environment for isolated crop activity. 4. Effects of hydrogen and hydroxyl ion concentrations were studied at pH values of 6.8, 7.5, 8.0, and 8.9. The highest average activity product, 1.011, was produced at a pH of about 8.0. 5. A satisfactory physiological salt solution for the isolated foregut of the American roach, Periplaneta americana

  8. The absence of rapid X-ray variability in active galaxies. Ph.D. Thesis - Maryland Univ.

    NASA Technical Reports Server (NTRS)

    Tennant, A. F.; Mushotzky, R. F.

    1982-01-01

    Variations on time scales ranging from minutes to several hours in the X-ray flux from 54 observations of 38 active galaxies are identified. The sample is composed mostly of Seyfert I galaxies but also includes radio galaxies, NELG's BL Lacs and 3C 273. Only NGC 6814 varied on time scales as short as 100 sec. No other source was observed to vary with a time scale of less than 12 hours. Large amplitude short term variations are not a characteristic of the X-ray emission from active galaxies. Upper limits on sigma sub I/I ranged from 2% for Cen A, 5% for NGC 4151, to approximately 20% for sources giving 1 ct/sec in the detector. Three objects NGC 3227, NGC 4151 and MCG 5-23-16 show variability consistent with a time scale of approximately 1 day. Ways to reconcile the rapid variability seen for NGC 6814 (and NGC 4051) with the general stability observed for the other objects are considered.

  9. Mutagenicity of organic pollutants adsorbed on suspended particulate matter in the center of Wrocław (Poland)

    NASA Astrophysics Data System (ADS)

    Bełcik, Maciej; Trusz-Zdybek, Agnieszka; Galas, Ewa; Piekarska, Katarzyna

    2014-10-01

    Mutagenicity of pollutants adsorbed on suspended dust of the PM10 fraction, collected in winter and summer season alike over the Wrocław city centre (Poland) was studied using the standard Salmonella assay (plate-incorporation) and the Kado modified assay (microsuspension method). The dust was collected using Staplex high volume air sampler. Further on it was extracted with dichloromethane in a Soxhlet apparatus. PAH content in extracts was determined by the high performance liquid chromatography technique using fluorescence detection, whereas the nitro-PAH content- by the gas chromatography using mass detection. Two Salmonella typhimurium strains, TA98 and YG1041, were used in the assays. The assays were conducted with and without a metabolic activation. Investigated air pollution extracts differed against each other with regard to a total content as well as to a percentage of individual compounds, depending on the sampling season. Both the total PAH content and the nitro-PAH content in the tested samples, and their spectrum as well, were found the highest in winter season. Higher mutagenic effect was noted for the dust extract from samples collected in wintertime than from those collected in summer. Pollutants directly affecting the genetic material and those showing such indirect action were present in the examined samples. The YG1041 strain turned out to be the most sensitive, which was the sign that large amounts of nitro-aromatic compounds were present in the tested samples. Obtained results proved that the Kado modified Salmonella assay would be useful for the atmospheric air pollution monitoring in urban agglomerations. Mutagenic effect in assays conducted according to the Kado procedure was obtained by using in the assays lower concentrations of tested extracts, compared to the classical assay.

  10. Mechanism of Flavoprotein l-6-Hydroxynicotine Oxidase: pH and Solvent Isotope Effects and Identification of Key Active Site Residues.

    PubMed

    Fitzpatrick, Paul F; Chadegani, Fatemeh; Zhang, Shengnan; Dougherty, Vi

    2017-02-14

    The flavoenzyme l-6-hydroxynicotine oxidase is a member of the monoamine oxidase family that catalyzes the oxidation of (S)-6-hydroxynicotine to 6-hydroxypseudooxynicotine during microbial catabolism of nicotine. While the enzyme has long been understood to catalyze oxidation of the carbon-carbon bond, it has recently been shown to catalyze oxidation of a carbon-nitrogen bond [Fitzpatrick, P. F., et al. (2016) Biochemistry 55, 697-703]. The effects of pH and mutagenesis of active site residues have now been utilized to study the mechanism and roles of active site residues. Asn166 and Tyr311 bind the substrate, while Lys287 forms a water-mediated hydrogen bond with flavin N5. The N166A and Y311F mutations result in ∼30- and ∼4-fold decreases in kcat/Km and kred for (S)-6-hydroxynicotine, respectively, with larger effects on the kcat/Km value for (S)-6-hydroxynornicotine. The K287M mutation results in ∼10-fold decreases in these parameters and a 6000-fold decrease in the kcat/Km value for oxygen. The shapes of the pH profiles are not altered by the N166A and Y311F mutations. There is no solvent isotope effect on the kcat/Km value for amines. The results are consistent with a model in which both the charged and neutral forms of the amine can bind, with the former rapidly losing a proton to a hydrogen bond network of water and amino acids in the active site prior to the transfer of hydride to the flavin.

  11. Changes in H+-ATP Synthase Activity, Proton Electrochemical Gradient, and pH in Pea Chloroplast Can Be Connected with Variation Potential

    PubMed Central

    Sukhov, Vladimir; Surova, Lyubov; Morozova, Ekaterina; Sherstneva, Oksana; Vodeneev, Vladimir

    2016-01-01

    Local stimulation induces generation and propagation of electrical signals, including the variation potential (VP) and action potential, in plants. Burning-induced VP changes the physiological state of plants; specifically, it inactivates photosynthesis. However, the mechanisms that decrease photosynthesis are poorly understood. We investigated these mechanisms by measuring VP-connected systemic changes in CO2 assimilation, parameters of light reactions of photosynthesis, electrochromic pigment absorbance shifts, and light scattering. We reveal that inactivation of photosynthesis in the pea, including inactivation of dark and light reactions, was connected with the VP. Inactivation of dark reactions decreased the rate constant of the fast relaxation of the electrochromic pigment absorbance shift, which reflected a decrease in the H+-ATP synthase activity. This decrease likely contributed to the acidification of the chloroplast lumen, which developed after VP induction. However, VP-connected decrease of the proton motive force across the thylakoid membrane, possibly, reflected a decreased pH in the stroma. This decrease may be another mechanism of chloroplast lumen acidification. Overall, stroma acidification can decrease electron flow through photosystem I, and lumen acidification induces growth of fluorescence non-photochemical quenching and decreases electron flow through photosystem II, i.e., pH decreases in the stroma and lumen, possibly, contribute to the VP-induced inactivation of light reactions of photosynthesis. PMID:27499760

  12. Effect of pH, aeration and sucrose feeding on the invertase activity of intact S. cerevisiae cells grown in sugarcane blackstrap molasses.

    PubMed

    Vitolo, M; Duranti, M A; Pellegrim, M B

    1995-08-01

    S. cerevisiae was grown in a blackstrap molasses containing medium in batch and fed-batch cultures. The following parameters were varied: pH (from 4.0 to 6.5), dissolved oxygen (DO) (from 0 to 5.0 mg O2 L-1) and sucrose feeding rate. When glucose concentration (S) was higher than 0.5 g L-1 a reduction in the specific invertase activity of intact cells (v) and an oscillatory behavior of v values during fermentation were observed. Both the invertase reduction and the oscillatory behavior of v values could be related to the glucose inhibitory effect on invertase biosynthesis. The best culture conditions for attaining S. cerevisiae cells suitable for invertase production were: temperature = 30 degrees C; pH = 5.0; DO = 3.3 mg O2 L-1; (S) = 0.5 g L-1 and sucrose added into the fermenter according to the equations: (V-Vo) = t2/16 or (V - Vo) = (Vf - Vo).(e0.6t-1)/10.

  13. Brown Kidney Bean Bowman-Birk Trypsin Inhibitor is Heat and pH Stable and Exhibits Anti-proliferative Activity.

    PubMed

    Chan, Yau Sang; Zhang, Yanbo; Ng, Tzi Bun

    2013-02-01

    A trypsin inhibitor with a molecular mass around 17 kDa was purified from the seeds of Phaseolus vulgaris cv. 'brown kidney bean'. The purification protocol involved, in sequence, affinity chromatography on Affi-gel blue gel, ion-exchange chromatography on Q-Sepharose and Mono Q, and gel filtration on Superdex 75. The molecular size and N-terminal amino acid sequence of the trypsin inhibitor resembled leguminous Bowman-Birk protease inhibitors (BBIs), signifying that brown kidney bean trypsin inhibitor is a BBI. Brown kidney bean trypsin inhibitor exhibited pronounced thermostability and pH stability. Its trypsin inhibitory activity was retained at all pH values (0-14) and up to 90 °C. The trypsin inhibitor also inhibited the proliferation of human breast cancer MCF7 cells with an IC(50) of 71.52 μM. On the other hand, it only slightly inhibited proliferation of hepatoma HepG2 and embryonic liver WRL68 cells at a concentration over 110 μM.

  14. Role of pH in the Formation of Structurally Stable and Catalytically Active TiO2-Supported Gold Catalysts

    SciTech Connect

    Veith, G.; Lupini, A; Dudney, N

    2008-01-01

    We report the investigation of titania (Degussa P25) supported gold catalysts prepared by magnetron sputtering. Catalysts grown on natural fumed titania were structurally unstable, resulting in the rapid coarsening of 2.4 nm gold clusters into large 20 nm gold clusters in a few days at room temperature under normal atmospheric conditions. However, treating the titania support powder to a mock deposition-precipitation process, at pH 4, followed by the subsequent deposition of gold onto this treated powder produced a remarkable enhancement in gold particle stability and a 20-fold enhancement of catalytic activity. Furthermore, it was found that treating the titania under basic conditions (pH 10) resulted in a further enhancement of structural stability and a further doubling of the reaction rate to 0.28 mol of CO/mol of Au s. This enhancement cannot be attributed to removing surface Cl- species from the titania, the formation of oxygen vacancies on the TiO2 surface, or an electronic effect. Instead, it appears to be associated with the formation of strongly bound hydroxyl species on the TiO2 surface. The formation of surface hydroxyls during the deposition-precipitation method is coincidental and contributes significantly to the properties of Au/TiO2 catalysts

  15. A factor confirmation and convergent validity of the “areas of worklife scale” (AWS) to Spanish translation

    PubMed Central

    2013-01-01

    Background Perceived incongruity between the individual and the job on work-life areas such as workload, control, reward, fairness, community and values have implications for the dimensions of burnout syndrome. The “Areas of Work-life Scale” (AWS) is a practical instrument to measure employees´ perceptions of their work environments. AIMS: Validate a Spanish translation of the AWS, and it relationship with Masclach Burnout Inventory dimensions. Methods The study was conducted in three medium-sized hospitals and seven rural and urban Primary Care centres (N = 871) in Spain. The “Maslach Burnout Inventory General Survey” (MBI-GS) and AWS were applied. We developed a complete psychometric analysis of its reliability, and validity. Results Data on the reliability supported a good internal consistency (Cronbach α between .71, and .85). Construct validity was confirmed by a six factor model of the AWS as a good measure of work environments (χ2(352) = 806.21, p < 0.001; χ2/df = 2.29; CFI = 0.935, RMSEA = 0.039); concurrent validity was analysed for its relationship with other measures (opposing dimensions to burnout, and MBI), and each correlation between dimensions and sub-dimensions were statistically significant; as well, predictive validity, by a series of Multiple Regression Analysis examined the resulting patterns of the Confirmatory Factor Analysis (CFA) confirms the relationship between the work-life areas and burnout dimensions. Conclusions Leiter and Maslach’s AWS has been an important instrument in exploring several work-life factors that contribute to burnout. This scale can now be used to assess the quality of work-life in order to design and assess the need for intervention programs in Spanish-speaking countries. PMID:23596987

  16. Effect of Mn-Zn ferrite on apatite-wollastonite glass-ceramic (A-W GC).

    PubMed

    Da Li, Guang; Zhou, Da Li; Pan, Tao Hua; Chen, Guo Sheng; Lin, Yun; Mao, Mao; Yan, Guo

    2009-08-01

    Magnetic bioactive glass-ceramics (M GC) were prepared by doping apatite-wollastonite glass-ceramic (A-W GC) with Mn-Zn ferrite. The effect of different contents of Mn-Zn ferrite on the phase structure, magnetic property and bioactivity of A-W GC was investigated. X-ray powder diffraction results showed that A-W GC exhibited apatite, fluorapatite and wollastonite as the main phases. The doping of Mn-Zn ferrite caused the formation of a new phase Zn(0.75)Mn(0.75)Fe(1.5)O(4) in M GC. The amount of this new phase increased with increasing content of Mn-Zn ferrite. Under a magnetic field of 7.96 x 10(5) A m(-1), the saturation magnetization of M GC increased from 4.63 to 9.7 A m(2) kg(-1), but the coercive forces of M GC decreased from 2.39 x 10(4) to 7.56 x 10(3) A m(-1) as the Mn-Zn ferrite content increased from 5% to 20% in the material. The bioactivity of samples was evaluated by soaking in simulated body fluid (SBF). The results showed that the doping of Mn-Zn ferrite decreased the bioactivity of A-W GC dramatically. It took 7 days for an apatite layer to form on the surface of A-W GC, while at least 30 days was needed for an apatite layer forming on the surface of M GC.

  17. Electrocatalytic activity of nickel oxide nanoparticles as mediatorless system for NADH and ethanol sensing at physiological pH solution.

    PubMed

    Sharifi, Ensiyeh; Salimi, Abdollah; Shams, Esmaeil

    2013-07-15

    The glassy carbon (GC) electrode modified by nickel oxide nanoparticles (NiOxNPs) is proposed as a novel electrocatalytic system for the oxidation of NADH without using any electron transfer mediator. Here, chronoamperometry was used not only as a simple method for the deposition of NiOxNPs onto the GC electrode but also as an efficient tool in the controlling of nanoparticles size and efficient electrocatalytic activity. The surface morphology and electrochemical properties of the NiOxNPs/GC electrode was investigated using scanning electron microscopy and cyclic voltammetry techniques, respectively. The NPs are deposited uniformly across the GC surface and the size of NiOxNPs varies from 20 to less than 100 nm. The NiOxNPs/GC electrode shows excellent electrocatalytic activity toward oxidation of NADH at reduced overvoltage. The detection limit and sensitivity of the modified electrode toward NADH were estimated to be 106 nM (S/N=3) and 0.052 μAμM(-1), respectively at a concentration range up to 1mM. Due to the biocompatibility of NiOxNPs toward biomolecules, this modified electrode can be used as an efficient transducer in the design of an ethanol biosensor based on the coupled alcohol dehydrogenase enzyme(ADH). Hydrodynamic amperometric detection of ethanol on the ADH-Nafion/NiOxNPs/GC modified electrode gives linear responses over the concentration range of 0.2-6mM with a detection limit of 6.4 μM and sensitivity of 36 nA mM(-1). Applicability of the proposed biosensor for ethanol detection in real samples, easy and simple preparation, being mediator free, high sensitivity and biocompatibility are the major advantages of the proposed biosensor.

  18. Interception of a Rh(I)-Rh(III) dinuclear trihydride complex revealing the dihydrogen activation by [Rh(CO)2{(R,R)-Ph-BPE}].

    PubMed

    Crozet, Delphine; McKay, David; Bijani, Christian; Gual, Aitor; Godard, Cyril; Claver, Carmen; Maron, Laurent; Urrutigoïty, Martine; Kalck, Philippe

    2012-03-28

    Reaction of [Rh(CO)(2){(R,R)-Ph-BPE}][BF(4)] 1 under 7 bar H(2) provides the dihydride [Rh(H)(2)(CO)(2){(R,R)-Ph-BPE}][BF(4)] 3, which reacts with the neutral hydride [Rh(H)(CO){(R,R)-Ph-BPE}] 2 arising from 3 in THF. The resulting complex is the dimeric monocationic Rh((I))-Rh((III)) complex [Rh(H)(2)(CO)(2){(R,R)-Ph-BPE}][BF(4)] 4.

  19. Acaricidal activity of Origanum bilgeri P.H. Davis (Lamiaceae) essential oil and its major component, carvacrol against adults Rhipicephalus turanicus (Acari: Ixodidae).

    PubMed

    Koc, Samed; Oz, Emre; Cinbilgel, Ilker; Aydin, Levent; Cetin, Huseyin

    2013-03-31

    The acaricidal activity of an essential oil obtained from aerial parts of Origanum bilgeri P.H. Davis (Lamiaceae), an endemic species in Turkey, and its major constituents, carvacrol was evaluated against unfed adults Rhipicephalus turanicus Pomerantzev (Acari: Ixodidae) collected from Kepez, Antalya. The composition of the essential oil was analyzed by GC/MS. The major compound identified in the oil was carvacrol (93.02%). Generally, tick mortalities to the O. bilgeri distillate and carvacrol increased with concentrations. O. bilgeri oil produced >83% mortality at 48h at a concentration of 0.8% and mortality was higher than 63% at a carvacrol concentration of 0.4%. Our results have shown that O. bilgeri essential oil and its major component, carvacrol, may have potential as acaricidal agents against R. turanicus.

  20. Purification, biochemical characterization and antifungal activity of a novel Aspergillus tubingensis glucose oxidase steady on broad range of pH and temperatures.

    PubMed

    Kriaa, Mouna; Hammami, Inès; Sahnoun, Mouna; Azebou, Manel Cheffi; Triki, Mohamed Ali; Kammoun, Radhouane

    2015-11-01

    This study was carried out to evaluate the in vitro and in vivo antifungal efficiency of Aspergillus tubingensis CTM 507 glucose oxidase (GOD) against plant pathogenic fungi. GOD displayed a wide inhibitory spectrum toward different fungi at a concentration of 20 AU. The GOD had a strong inhibitor effect on mycelia growth and spore germination of Pythium ultimum. Interestingly, the GOD exhibited a potent in vivo antifungal effect against P. ultimum responsible for potato plants disease. The antifungal GOD was purified 13-fold with 27 % yield and a specific activity of 3435 U/mg. The relative molecular mass of the GOD was 180 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The GOD activity was optimum at pH 4.5 and 60 °C. It was found to be stable over a large pH range (3-9). It also displayed a marked thermostability with a 50-min half-life at 65 °C. The 10 residues of the N-terminal sequence of the purified GOD (S-K-G-S-A-V-T-T-P-D) showed no homology to the other reported GOD, identifying a novel GOD. FTIR spectroscopic analysis revealed the presence of C-O and C=O groups corresponding to a D-glucono-lactone. The findings indicated that GOD is the first A. tubingensis-produced fungicide ever reported to exhibit such promising biological properties. It could become a natural alternative to synthetic fungicides to control certain important plant microbial diseases.

  1. Phosphorylation of ribosomal proteins during meiotic maturation and following activation in starfish oocytes: its relationship with changes of intracellular pH.

    PubMed

    Peaucellier, G; Picard, A; Robert, J J; Capony, J P; Labbe, J C; Doree, M

    1988-01-01

    An increased phosphorylation of ribosomal protein S6 has been shown to be correlated with an increase of intracellular pH (pHi) and with stimulation of protein synthesis in many systems. In this research changes in ribosome phosphorylation following hormone-induced meiotic maturation and fertilization or activation by ionophore A23187 were investigated in starfish oocytes. The hormone was found to stimulate, even in the absence of external Na+, the phosphorylation on serine residues of an Mr 31,000 protein identified as S6, as well as that of an acidic Mr 47,000 protein, presumably S1, on threonine residues. Phosphorylation of ribosomes was an early consequence of hormonal stimulation and did not decrease after completion of meiotic maturation. Fertilization or activation by ionophore of prophase-arrested oocytes also stimulated ribosome phosphorylation. Only S6 was labeled in this case, but to a lesser extent than upon hormone-induced meiotic maturation. Changes in pHi were monitored with ion-specific microelectrodes throughout meiotic maturation and following either fertilization or activation. The pHi did not change before germinal vesicle breakdown (GVBD) following hormone addition, but it increased before first polar body emission. It also increased following fertilization or activation by ionophore or the microinjection of Ca-EGTA. In all cases, alkalinization did not depend on activation of an amiloride-sensitive Na+/H+ exchanger. Microinjection of an alkaline Hepes buffer or external application of ammonia, both of which increased pHi, prevented unfertilized oocytes from arresting after formation of the female pronucleus and induced chromosome cycling. Phosphorylation of S6 was still observed following fertilization or induction of maturation when pHi was decreased by external application of acetate, a treatment which suppressed the emission of polar bodies. Protein synthesis increased in prophase-arrested oocytes after fertilization or activation. It also

  2. Final Report for the Erosion-Corrosion Anaysis of Tank 241-AW-02E Feed Pump Pit Jumpers B-2 and 1-4 Removed from Service in 2013

    SciTech Connect

    Page, Jason S.

    2014-04-07

    This document is the final report summarizing the results in the examination of two pipe sections (jumpers) from the tank 241-AW-02E feed pump pit in the 241-AW tank farm. These pipe section samples consisted of jumper AW02E-WT-J-[B – 2] and jumper AW02E-WT-J-[1 – 4]. For the remainder of this report, these jumpers will be referred to as B – 2 and 1 – 4.

  3. Mechanism of phosphoryl transfer by nucleoside diphosphate kinase pH dependence and role of the active site Lys16 and Tyr56 residues.

    PubMed

    Schneider, B; Babolat, M; Xu, Y W; Janin, J; Véron, M; Deville-Bonne, D

    2001-04-01

    Nucleoside diphosphate (NDP) kinase phosphorylates nucleoside diphosphates with little specificity for the base and the sugar. Although nucleotide analogues used in antiviral therapies are also metabolized to their triphosphate form by NDP kinase, their lack of the 3'-hydroxyl of the ribose, which allows them to be DNA chain terminators, severely impairs the catalytic efficiency of NDP kinase. We have analyzed the kinetics parameters of several mutant NDP kinases modified on residues (Lys16, Tyr56, Asn119) interacting with the gamma-phosphate and/or the 3'-OH of the Mg2+-ATP substrate. We compared the relative contributions of the active-site residues and the substrate 3'-OH for point mutations on Lys16, Tyr56 and Asn119. Analysis of additional data from pH profiles identify the ionization state of these residues in the enzyme active form. X-ray structure of K16A mutant NDP kinase shows no detectable rearrangement of the residues of the active site.

  4. Iron chelating active packaging: Influence of competing ions and pH value on effectiveness of soluble and immobilized hydroxamate chelators.

    PubMed

    Ogiwara, Yoshiko; Roman, Maxine J; Decker, Eric A; Goddard, Julie M

    2016-04-01

    Many packaged foods utilize synthetic chelators (e.g. ethylenediaminetetraacetic acid, EDTA) to inhibit iron-promoted oxidation or microbial growth which would result in quality loss. To address consumer demands for all natural products, we have previously developed a non-migratory iron chelating active packaging material by covalent immobilization of polyhydroxamate and demonstrated its efficacy in delaying lipid oxidation. Herein, we demonstrate the ability of this hydroxamate-functionalized iron chelating active packaging to retain iron chelating capacity; even in the presence of competing ions common in food. Both immobilized and soluble hydroxamate chelators retained iron chelating capacity in the presence of calcium, magnesium, and sodium competing ions, although at pH 5.0 the presence of calcium reduced immobilized hydroxamate iron chelation. A strong correlation was found between colorimetric and mass spectral analysis of iron chelation by the chelating packaging material. Such chelating active packaging may support reducing additive use in product formulations, while retaining quality and shelf life.

  5. Different rate-limiting activities of intracellular pH regulators for HCO3(-) secretion stimulated by forskolin and carbachol in rat parotid intralobular ducts.

    PubMed

    Ueno, Kaori; Hirono, Chikara; Kitagawa, Michinori; Shiba, Yoshiki; Sugita, Makoto

    2016-11-01

    Intracellular pH (pHi) regulation fundamentally participates in maintaining HCO3(-) release from HCO3(-)-secreting epithelia. We used parotid intralobular ducts loaded with BCECF to investigate the contributions of a carbonic anhydrase (CA), anion channels and a Na(+)-H(+) exchanger (NHE) to pHi regulation for HCO3(-) secretion by cAMP and Ca(2+) signals. Resting pHi was dispersed between 7.4 and 7.9. Forskolin consistently decreased pHi showing the dominance of pHi-lowering activities, but carbachol gathered pHi around 7.6. CA inhibition suppressed the forskolin-induced decrease in pHi, while it allowed carbachol to consistently increase pHi by revealing that carbachol prominently activated NHE via Ca(2+)-calmodulin. Under NHE inhibition, forskolin and carbachol induced the remarkable decreases in pHi, which were slowed predominantly by CA inhibition and by CA or anion channel inhibition, respectively. Our results suggest that forskolin and carbachol primarily activate the pHi-lowering CA and pHi-raising NHE, respectively, to regulate pHi for HCO3(-) secretion.

  6. Tank 214-AW-105, grab samples, analytical results for the finalreport

    SciTech Connect

    Esch, R.A.

    1997-02-20

    This document is the final report for tank 241-AW-105 grab samples. Twenty grabs samples were collected from risers 10A and 15A on August 20 and 21, 1996, of which eight were designated for the K Basin sludge compatibility and mixing studies. This document presents the analytical results for the remaining twelve samples. Analyses were performed in accordance with the Compatibility Grab Sampling and Analysis Plan (TSAP) and the Data Quality Objectives for Tank Farms Waste Compatibility Program (DO). The results for the previous sampling of this tank were reported in WHC-SD-WM-DP-149, Rev. 0, 60-Day Waste Compatibility Safety Issue and Final Results for Tank 241-A W-105, Grab Samples 5A W-95-1, 5A W-95-2 and 5A W-95-3. Three supernate samples exceeded the TOC notification limit (30,000 microg C/g dry weight). Appropriate notifications were made. No immediate notifications were required for any other analyte. The TSAP requested analyses for polychlorinated biphenyls (PCB) for all liquids and centrifuged solid subsamples. The PCB analysis of the liquid samples has been delayed and will be presented in a revision to this document.

  7. From eugenics to lysenkoism: the evolution of Stanisław Skowron.

    PubMed

    Dejong-Lambert, William

    2009-01-01

    This article describes the relationship between Polish geneticist Stanisław Skowron's views on eugenics during the interwar period, his experiences in Nazi concentration camps during World War II, and his response to Trofim D. Lysenko's ban on genetic research in Soviet-allied states after 1948. Skowron was educated at the Jagiellonian University in Krakow and received funding from the Rockefeller Foundation to study in the United States, Italy, Denmark, and Great Britain from 1924 to 1926. His exposure to research being conducted outside of Poland made him an important figure in Polish genetics. During this time Skowron also began to believe that an understanding of biological principles of heredity could play an important role in improving Polish society and became a supporter of eugenics. In 1939 he was arrested along with other faculty members at the Jagiellonian and sent to Sachsenhausen and Dachau. In 1947 he published the first book updating Polish biologists on recent developments in genetics; however, after learning of the outcome of the 1948 session of the Lenin All-Union Academy of Agricultural Sciences in Moscow, Skowron emerged as on of the most vocal advocates for Michurinism. I argue that Skowron's conversion to Lysenkoism was motivated by more than fear or opportunism, and is better understood as the product of his need to rationalize his own support for a theory he could not possibly have believed was correct.

  8. Orographic Flows across the Antarctic Peninsula: a comparison of AWS data and regional atmospheric model simulations.

    NASA Astrophysics Data System (ADS)

    Kirchgaessner, A.

    2012-04-01

    Over the past 50 years the Antarctic Peninsula (AP) has been one of the most rapidly warming regions on the planet, with the strongest warming rate occurring in winter along the west coast of the Peninsula. In summer, the largest temperature increase is observed over the northern part of the east coast. During summer the temperature on both sides of the Peninsula is correlated with the circumpolar westerlies, expressed in the Southern Annular Mode (SAM). The suggestion is that the stronger westerlies interact with the orographic barrier formed by the Peninsula, resulting in Föhn events on the lee side in the area of the Larsen Ice Shelf. In this study we use standard meteorological parameters as measured by Automatic Weather Stations (AWS) on either side of the Antarctic Peninsula to derive an indicator for Föhn events. This indicator is then used to analyse simulations by two high-resolution regional atmospheric models - the WRF model, run at 5 km resolution as part of the Antarctic Mesoscale Prediction System (AMPS) and the UK Met Office Unified Model (UM) run at 4 km resolution as part of an experimental real-time forecast system for the Antarctic Peninsula region with regard to their representation of such events. In a further step we will look into which impact the models' performance with regard to Föhn events may have on simulated components of the surface energy balance and thus potential melt.

  9. Novel silica surface charge density mediated control of the optical properties of embedded optically active materials and its application for fiber optic pH sensing at elevated temperatures.

    PubMed

    Wang, Congjun; Ohodnicki, Paul R; Su, Xin; Keller, Murphy; Brown, Thomas D; Baltrus, John P

    2015-02-14

    Silica and silica incorporated nanocomposite materials have been extensively studied for a wide range of applications. Here we demonstrate an intriguing optical effect of silica that, depending on the solution pH, amplifies or attenuates the optical absorption of a variety of embedded optically active materials with very distinct properties, such as plasmonic Au nanoparticles, non-plasmonic Pt nanoparticles, and the organic dye rhodamine B (not a pH indicator), coated on an optical fiber. Interestingly, the observed optical response to varying pH appears to follow the surface charge density of the silica matrix for all the three different optically active materials. To the best of our knowledge, this optical effect has not been previously reported and it appears universal in that it is likely that any optically active material can be incorporated into the silica matrix to respond to solution pH or surface charge density variations. A direct application of this effect is for optical pH sensing which has very attractive features that can enable minimally invasive, remote, real time and continuous distributed pH monitoring. Particularly, as demonstrated here, using highly stable metal nanoparticles embedded in an inorganic silica matrix can significantly improve the capability of pH sensing in extremely harsh environments which is of increasing importance for applications in unconventional oil and gas resource recovery, carbon sequestration, water quality monitoring, etc. Our approach opens a pathway towards possible future development of robust optical pH sensors for the most demanding environmental conditions. The newly discovered optical effect of silica also offers the potential for control of the optical properties of optically active materials for a range of other potential applications such as electrochromic devices.

  10. Effect of concentrate level and feeding management on chewing activities, saliva production, and ruminal pH of lactating dairy cows.

    PubMed

    Maekawa, M; Beauchemin, K A; Christensen, D A

    2002-05-01

    Eight ruminally cannulated lactating Holstein cows were used in a double 4 x 4 Latin square to determine the effects of 1) proportion of barley silage [40, 50, and 60% of dry matter (DM)] in the diet, and 2) feeding a total mixed ration (TMR) compared with separate ingredients (SI) on chewing activities, saliva production, and ruminal pH. Although cows fed SI were offered a diet containing 50% silage, they actually consumed a diet containing 43% silage (DM basis). Dry matter intake and milk yield were similar for all diets (18.2 kg of DM/d and 27.2 kg/d, respectively). Cows fed the 40% silage TMR spent more time eating than cows fed SI (243 vs. 198 min/d), but rumination time was similar (546 min/d). Eating time was similar among the TMR diets, but rumination time increased from 498 to 516 and 584 min/d as silage in the TMR increased from 40 to 50, and then to 60%, respectively. The secretion of saliva per gram of feed was 4.43, 3.18, and 1.19 ml/g of DM with consumption of silage, TMR, and concentrate, respectively. Resting salivation rate was similar for all diets (101 ml/min). Regardless of the diet, cows secreted 239 +/- 17 L/d of saliva, and ruminal pH was below 5.8 for 10 h/d. Results indicated increased chewing time did not increase total daily saliva secretion because increased eating and ruminating saliva was associated with decreased resting saliva. Feeding SI increased the risk of acidosis, because cows ate a higher proportion of concentrate than intended.

  11. A Determination and Comparison of Urease Activity in Feces and Fresh Manure from Pig and Cattle in Relation to Ammonia Production and pH Changes

    PubMed Central

    Dai, Xiaorong; Karring, Henrik

    2014-01-01

    Ammonia emission from animal production is a major environmental problem and has impacts on the animal health and working environment inside production houses. Ammonia is formed in manure by the enzymatic degradation of urinary urea and catalyzed by urease that is present in feces. We have determined and compared the urease activity in feces and manure (a urine and feces mixture) from pigs and cattle at 25°C by using Michaelis-Menten kinetics. To obtain accurate estimates of kinetic parameters Vmax and K'm, we used a 5 min reaction time to determine the initial reaction velocities based on total ammoniacal nitrogen (TAN) concentrations. The resulting Vmax value (mmol urea hydrolyzed per kg wet feces per min) was 2.06±0.08 mmol urea/kg/min and 0.80±0.04 mmol urea/kg/min for pig feces and cattle feces, respectively. The K'm values were 32.59±5.65 mmol urea/l and 15.43±2.94 mmol urea/l for pig feces and cattle feces, respectively. Thus, our results reveal that both the Vmax and K'm values of the urease activity for pig feces are more than 2-fold higher than those for cattle feces. The difference in urea hydrolysis rates between animal species is even more significant in fresh manure. The initial velocities of TAN formation are 1.53 mM/min and 0.33 mM/min for pig and cattle manure, respectively. Furthermore, our investigation shows that the maximum urease activity for pig feces occurs at approximately pH 7, and in cattle feces it is closer to pH 8, indicating that the predominant fecal ureolytic bacteria species differ between animal species. We believe that our study contributes to a better understanding of the urea hydrolysis process in manure and provides a basis for more accurate and animal-specific prediction models for urea hydrolysis rates and ammonia concentration in manures and thus can be used to predict ammonia volatilization rates from animal production. PMID:25397404

  12. A determination and comparison of urease activity in feces and fresh manure from pig and cattle in relation to ammonia production and pH changes.

    PubMed

    Dai, Xiaorong; Karring, Henrik

    2014-01-01

    Ammonia emission from animal production is a major environmental problem and has impacts on the animal health and working environment inside production houses. Ammonia is formed in manure by the enzymatic degradation of urinary urea and catalyzed by urease that is present in feces. We have determined and compared the urease activity in feces and manure (a urine and feces mixture) from pigs and cattle at 25°C by using Michaelis-Menten kinetics. To obtain accurate estimates of kinetic parameters Vmax and K'm, we used a 5 min reaction time to determine the initial reaction velocities based on total ammoniacal nitrogen (TAN) concentrations. The resulting Vmax value (mmol urea hydrolyzed per kg wet feces per min) was 2.06±0.08 mmol urea/kg/min and 0.80±0.04 mmol urea/kg/min for pig feces and cattle feces, respectively. The K'm values were 32.59±5.65 mmol urea/l and 15.43±2.94 mmol urea/l for pig feces and cattle feces, respectively. Thus, our results reveal that both the Vmax and K'm values of the urease activity for pig feces are more than 2-fold higher than those for cattle feces. The difference in urea hydrolysis rates between animal species is even more significant in fresh manure. The initial velocities of TAN formation are 1.53 mM/min and 0.33 mM/min for pig and cattle manure, respectively. Furthermore, our investigation shows that the maximum urease activity for pig feces occurs at approximately pH 7, and in cattle feces it is closer to pH 8, indicating that the predominant fecal ureolytic bacteria species differ between animal species. We believe that our study contributes to a better understanding of the urea hydrolysis process in manure and provides a basis for more accurate and animal-specific prediction models for urea hydrolysis rates and ammonia concentration in manures and thus can be used to predict ammonia volatilization rates from animal production.

  13. A Comparison of the AFGL Flash, Draper Dart and AWS Haze Models with the Rand Wetta Model for Calculating Atmospheric Contrast Reduction.

    DTIC Science & Technology

    1982-03-01

    AD-AI1S 425 AIR FORCE ENVIRON0ENTAL TECHNICAL APPLICATIONS CENTER--ETC F/G 4/2 A COMPARISON OF THE AFOL FLASH, DRAPER DART AND AWS HAZE MODELS--ETC(U...COMPARISON OF THE AFGL FLASH, DRAPER DART AND AWS HAZE MODELS WITH THE RAND WETTA MODEL FOR CALCULATING ATMOSPHERIC CONTRAST REDUCTION BY1 DR. PATRICK J...AFOL FLASH, DRAPER DART AND AWS HAE MODELS WITH THE RAND WETTA MODEL FOR CALCULATING ATMOIPlURIC CONTRAST REDUCTION, March 1982, is approved for public

  14. Zn(2+) induces hyperpolarization by activation of a K(+) channel and increases intracellular Ca(2+) and pH in sea urchin spermatozoa.

    PubMed

    Beltrán, Carmen; Rodríguez-Miranda, Esmeralda; Granados-González, Gisela; García de De la Torre, Lucia; Nishigaki, Takuya; Darszon, Alberto

    2014-10-01

    Zinc (Zn(2+)) has been recently recognized as a crucial element for male gamete function in many species although its detailed mechanism of action is poorly understood. In sea urchin spermatozoa, Zn(2+) was reported as an essential trace ion for efficient sperm motility initiation and the acrosome reaction by modulating intracellular pH (pHi). In this study we found that submicromolar concentrations of free Zn(2+) change membrane potential (Em) and increase the concentration of intracellular Ca(2+) ([Ca(2+)]i) and cAMP in Lytechinus pictus sperm. Our results indicate that the Zn(2+) response in sperm of this species mainly involves an Em hyperpolarization caused by K(+) channel activation. The pharmacological profile of the Zn(2+)-induced hyperpolarization indicates that the cGMP-gated K(+) selective channel (tetraKCNG/CNGK), which is crucial for speract signaling, is likely a main target for Zn(2+). Considering that Zn(2+) also induces [Ca(2+)]i fluctuations, our observations suggest that Zn(2+) activates the signaling cascade of speract, except for an increase in cGMP, and facilitates sperm motility initiation upon spawning. These findings provide new insights about the role of Zn(2+) in male gamete function.

  15. Characterisation of selected active agents regarding pKa values, solubility concentrations and pH profiles by SiriusT3.

    PubMed

    Schönherr, D; Wollatz, U; Haznar-Garbacz, D; Hanke, U; Box, K J; Taylor, R; Ruiz, R; Beato, S; Becker, D; Weitschies, W

    2015-05-01

    The aim of this work was to determine pKa values and solubility properties of 34active agents using the SiriusT3 apparatus. The selected drug substances belong to the groups of ACE-inhibitors, β-blockers, antidiabetics and lipid lowering substances. Experimentally obtained pKa and intrinsic solubility values were compared to calculated values (program ACD/ChemSketch) and pKa values to published data as well. Solubility-pH profiles were generated to visualise the substance solubility over the gastrointestinal pH range. The relationship between the solubility characteristic of a substance, its bioavailability and categorisation according to the Biopharmaceutics Classification System (BCS) was examined as well. The results showed a good agreement between experimentally obtained, calculated and published pKa values. The measured and calculated intrinsic solubility values indicated several major deviations. All solubility-pH profiles showed the expected shape and appearance for acids, bases or zwitterionic substances. The obtained results for the pKa and solubility measurements of the examined active agents may help to predict their physicochemical behaviour in vivo, and to understand the bioavailability of the substances according to their BCS categorisation. The easy and reproducible determination of pKa and solubility values makes the SiriusT3 apparatus a useful tool in early stages of drug and formulation development.

  16. Restoration of blood pH between repeated bouts of high-intensity exercise: effects of various active-recovery protocols.

    PubMed

    Del Coso, Juan; Hamouti, Nassim; Aguado-Jimenez, Roberto; Mora-Rodriguez, Ricardo

    2010-02-01

    To determine which active-recovery protocol would reduce faster the high blood H(+) and lactate concentrations produced by repeated bouts of high-intensity exercise (HIE). On three occasions, 11 moderately trained males performed 4 bouts (1.5 min) at 163% of their respiratory compensation threshold (RCT) interspersed with active-recovery: (1) 4.5 min pedalling at 24% RCT (S(HORT)); (2) 6 min at 18% RCT (M(EDIUM)); (3) 9 min at 12% RCT (L(ONG)). The total work completed during recovery was the same in all three trials. Respiratory gases and arterialized-blood samples were obtained during exercise. At the end of exercise, L(ONG) in comparison to S(HORT) and M(EDIUM) increased plasma pH (7.32 +/- 0.02 vs. approximately 7.22 +/- 0.03; P < 0.05), while reduced lactate concentration (8.5 +/- 0.9 vs. approximately 10.9 +/- 0.8 mM; P < 0.05). Ventilatory equivalent for CO(2) was higher in L(ONG) than S(HORT) and M(EDIUM) (31.4 +/- 0.5 vs. approximately 29.6 +/- 0.5; P < 0.05). Low-intensity prolonged recovery between repeated bouts of HIE maximized H(+) and lactate removal likely by enhancing CO(2) unloading.

  17. Biodegradation of Medium Chain Hydrocarbons by Acinetobacter venetianus 2AW Immobilized to Hair-Based Adsorbent Mats (Postprint)

    DTIC Science & Technology

    2010-09-01

    Difco Lab- oratories, Detroit, MI) was used as a complete medium for bacterial cultivation . Bushnell-Haas (BH) (Becton Dickinson 3 Laboratories, Sparks...Bacterial cultivation and characterization Acinetobacter venetianus 2AW was cultured aerobically in BH broth (Becton Dickinson Laboratories...over a period of 40 min using a Synergy 4 hybrid plate reader (Biotek, Winooski, VT). The same cell dilutions were cultivated overnight at room

  18. The type IIP supernova 2012aw in M95: Hydrodynamical modeling of the photospheric phase from accurate spectrophotometric monitoring

    SciTech Connect

    Dall'Ora, M.; Botticella, M. T.; Della Valle, M.; Pumo, M. L.; Zampieri, L.; Tomasella, L.; Cappellaro, E.; Benetti, S.; Pignata, G.; Bufano, F.; Bayless, A. J.; Pritchard, T. A.; Taubenberger, S.; Benitez, S.; Kotak, R.; Inserra, C.; Fraser, M.; Elias-Rosa, N.; Haislip, J. B.; Harutyunyan, A.; and others

    2014-06-01

    We present an extensive optical and near-infrared photometric and spectroscopic campaign of the Type IIP supernova SN 2012aw. The data set densely covers the evolution of SN 2012aw shortly after the explosion through the end of the photospheric phase, with two additional photometric observations collected during the nebular phase, to fit the radioactive tail and estimate the {sup 56}Ni mass. Also included in our analysis is the previously published Swift UV data, therefore providing a complete view of the ultraviolet-optical-infrared evolution of the photospheric phase. On the basis of our data set, we estimate all the relevant physical parameters of SN 2012aw with our radiation-hydrodynamics code: envelope mass M {sub env} ∼ 20 M {sub ☉}, progenitor radius R ∼ 3 × 10{sup 13} cm (∼430 R {sub ☉}), explosion energy E ∼ 1.5 foe, and initial {sup 56}Ni mass ∼0.06 M {sub ☉}. These mass and radius values are reasonably well supported by independent evolutionary models of the progenitor, and may suggest a progenitor mass higher than the observational limit of 16.5 ± 1.5 M {sub ☉} of the Type IIP events.

  19. Influence of Process Parameters on the Quality of Aluminium Alloy EN AW 7075 Using Selective Laser Melting (SLM)

    NASA Astrophysics Data System (ADS)

    Kaufmann, N.; Imran, M.; Wischeropp, T. M.; Emmelmann, C.; Siddique, S.; Walther, F.

    Selective laser melting (SLM) is an additive manufacturing process, forming the desired geometry by selective layer fusion of powder material. Unlike conventional manufacturing processes, highly complex parts can be manufactured with high accuracy and little post processing. Currently, different steel, aluminium, titanium and nickel-based alloys have been successfully processed; however, high strength aluminium alloy EN AW 7075 has not been processed with satisfying quality. The main focus of the investigation is to develop the SLM process for the wide used aluminium alloy EN AW 7075. Before process development, the gas-atomized powder material was characterized in terms of statistical distribution: size and shape. A wide range of process parameters were selected to optimize the process in terms of optimum volume density. The investigations resulted in a relative density of over 99%. However, all laser-melted parts exhibit hot cracks which typically appear in aluminium alloy EN AW 7075 during the welding process. Furthermore the influence of processing parameters on the chemical composition of the selected alloy was determined.

  20. Influence of pH on the formulation of TiO{sub 2} powder prepared by co-precipitation of TiCl{sub 3} and photocatalytic activity

    SciTech Connect

    Yudoyono, Gatut Zharvan, Vicran; Ichzan, Nur; Daniyati, Rizqa; Indarto, Bachtera; Pramono, Yono Hadi; Zainuri, Mochamad; Darminto

    2016-02-08

    Titanium dioxide (titania) nanoparticle were synthesized by coprecipitation process of titanium trichloride (TiCl{sub 3}) in aqueous medium, with NH{sub 4}OH as pH regulator. The pH solution was varied during the synthesis process between pH 3-8.4, and all samples were calcined temperature at 400°C for 3 hours. Characteristics and properties of the TiO{sub 2} powder were investigated using X-ray diffraction (XRD), Scanning Electron Microscopy (SEM). XRD results show that the single-phase rutile formed when the pH is less than 5, anatase single phase formed began pH 8, and the pH of the solution between 5-8 formed mixed phase rutile-anatase-brookite, rutile-brookite or anatase-brookite. Methylene Blue (MB) photodegradation test were performed in order to evaluate photocatalytic activity. Nanoparticles TiO{sub 2} rutile, anatase phase, and mixed phase rutile-brookite, anatase-brookite used to test the photocatalytic activity by measuring the absorbance spectrum photodegradation using UV-Vis spectrometer. The test results showed that the mixture phase of rutile-brookite provide the greatest photodegradation than other phases.

  1. Reducing activity, glucose metabolism and acid tolerance response of Bacillus cereus grown at various pH and oxydo-reduction potential levels.

    PubMed

    Le Lay, Julien; Bahloul, Halim; Sérino, Sylvie; Jobin, Michel; Schmitt, Philippe

    2015-04-01

    Bacillus cereus is a major foodborne bacterial pathogen able to survive a large number of physical-chemical stresses. B. cereus encounters different pH and redox potential (Eh7) levels during its passage through the gastrointestinal tract. Analysis of the combined influence of pH and redox stresses on B. cereus F4430/73 physiology found that B. cereus F4430/73 growth at pH 7.0 at 37 °C had strong reducing capacities, with a total change of 315 mV from an initial redox value of +214 ± 17 mV. The combination of low Eh7 and low pH led to a drastic reduction of growth parameters compared to oxidative Eh7 and neutral pH. Metabolic analysis showed that low pH significantly modifies glucose fermentative metabolism, with changes including decreased production of acid metabolite (acetate, lactate, formate) and increased production of 2,3-butanediol. Low Eh7 slightly enhanced the acid-tolerance response of B. cereus whereas low pH pre-adaptation led to thermal stress cross-protection. These results highlight new mechanisms that bring fresh insight into B. cereus pH and redox stress adaptations.

  2. THE RED SUPERGIANT PROGENITOR OF SUPERNOVA 2012aw (PTF12bvh) IN MESSIER 95

    SciTech Connect

    Van Dyk, Schuyler D.; and others

    2012-09-10

    We report on the direct detection and characterization of the probable red supergiant (RSG) progenitor of the intermediate-luminosity Type II-Plateau (II-P) supernova (SN) 2012aw in the nearby (10.0 Mpc) spiral galaxy Messier 95 (M95; NGC 3351). We have identified the star in both Hubble Space Telescope images of the host galaxy, obtained 17-18 yr prior to the explosion, and near-infrared ground-based images, obtained 6-12 yr prior to the SN. The luminous supergiant showed evidence for substantial circumstellar dust, manifested as excess line-of-sight extinction. The effective total-to-selective ratio of extinction to the star was R'{sub V} Almost-Equal-To 4.35, which is significantly different from that of diffuse interstellar dust (i.e., R{sub V} = 3.1), and the total extinction to the star was therefore, on average, A{sub V} Almost-Equal-To 3.1 mag. We find that the observed spectral energy distribution for the progenitor star is consistent with an effective temperature of 3600 K (spectral type M3), and that the star therefore had a bolometric magnitude of -8.29. Through comparison with recent theoretical massive-star evolutionary tracks we can infer that the RSG progenitor had an initial mass 15 {approx}< M{sub ini}(M{sub Sun }) < 20. Interpolating by eye between the available tracks, we surmise that the star had initial mass {approx}17-18 M{sub Sun }. The circumstellar dust around the progenitor must have been destroyed in the explosion, as the visual extinction to the SN is found to be low (A{sub V} = 0.24 mag with R{sub V} = 3.1).

  3. Investigation of solubilising effects of bile salts on an active pharmaceutical ingredient with unusual pH dependent solubility by NMR spectroscopy.

    PubMed

    Vogtherr, M; Marx, A; Mieden, A-C; Saal, C

    2015-05-01

    The interaction between an ampholytic and amphiphilic Active Pharmaceutical Ingredient (API) showing unusual pH dependent solubility and Fasted State Simulated Intestinal Fluid (FaSSIF) was studied by NMR spectroscopy. Solubility in FaSSIF was drastically increased, about 30 fold, compared to simulated gastrointestinal fluid without bile salts. Our studies aimed at understanding the mechanisms that lead to this drastic enhancement. All species present in solution at various concentrations of API were characterised by Diffusion Ordered Spectroscopy (DOSY) NMR measurements. These indicated the presence of mixed taurocholate-lecithin and pure taurocholate micelles in pure FaSSIF, and formation of mixed taurocholate-API micelles after addition of API. The formation of taurocholate-API micelles was also supported by Nuclear Overhauser Effect/Enhancement (NOE) contacts between taurocholate and the API. Formation of mixed taurocholate-API micelles took place at the expense of pure taurocholate micelles, whereas mixed taurocholate-lecithin micelles remained uninfluenced by the presence of API. Our results showed that the increase in solubility was due to similar amphiphilic properties of the API and taurocholate which enabled formation of mixed taurocholate-API micelles. From results of determination of solubility as well as NMR experiments a phase diagram comprising several micellar species was derived.

  4. A Computational Study of the Abrupt Wing Stall (AWS) Characteristics for Various Fighter Jets. Part 1; F/A-18E and F-16C

    NASA Technical Reports Server (NTRS)

    Parikh, Paresh; Chung, James

    2003-01-01

    Steady state Computational Fluid Dynamic (CFD) simulations are used to gain an understanding of the physics behind the Abrupt Wing Stall (AWS) phenomenon and to arrive at static Figures of Merit (FOMs). Navier-Stokes simulations are conducted using the NASA Langley developed TetrUSS simulation suite which is based on tetrahedral, unstructured grids. The physics of the AWS phenomenon is understood by comparing CFD simulation results on two aircraft; a pre-production F/A-18E configuration which exhibits AWS phenomenon under certain geometric and flow conditions, and an F-16C aircraft configuration that does not. The CFD code is first validated against two sets of experimental data to build confidence in its use for the problem of AWS. An attempt is then made to understand the possible causes of AWS by analyzing and comparing the detailed flow fields between the two configurations under a variety of flow conditions. Based on this approach, a number of static Figures of Merit are developed to predict the potential existence of AWS. The FOMs include the break in the lift and wing root bending moment versus angle-of-attack (AoA) curves and the rate of change of sectional lift with respect to AoA. A companion paper, in Part II, describes a similar CFD study on two other aircraft, the AV-8B Harrier and F/A-18C. Results from both these studies, as well as other CFD studies conducted as part of the AWS program are used to recommend a CFD procedure for predicting the existence of AWS in future aerospace designs.

  5. Kinetic activity, membrane mitochondrial potential, lipid peroxidation, intracellular pH and calcium of frozen/thawed bovine spermatozoa treated with metabolic enhancers.

    PubMed

    Boni, R; Gallo, A; Cecchini, S

    2017-01-01

    Owing to the progressive decline of sperm motility during storage there is a need to find substances capable of enhancing sperm energy metabolism and motility and/or preserving it from oxidative damage. The aim of this study was to evaluate in frozen/thawed bovine spermatozoa the effect of several compounds, such as myo-inositol, pentoxifylline, penicillamine + hypotaurine + epinephrine mixture (PHE), caffeine and coenzyme Q10+ zinc + d-aspartate mixture (CZA), on either kinetic or metabolic parameters. Sperm kinetics was evaluated by Sperm Class Analyser whereas specific fluorochromes were used to evaluated mitochondrial membrane potential (MMP), intracellular pH, intracellular calcium concentration and lipid peroxidation. Lipid peroxidation was also evaluated by TBARS analysis. Treatments significantly affected total and progressive motility with different dynamics in relation to the incubation time. After the first hour of incubation, CZA treatment produced the best performance in total and progressive sperm motility as well as in curvilinear velocity, average path velocity and amplitude of head displacement, whereas pentoxifylline stimulated the highest straight-line velocity. MMP showed higher values (p < 0.01) after treatment with pentoxifylline and PHE. Intracytoplasmic calcium concentration and lipid peroxidation were significantly (p < 0.05) affected by the incubation time rather than the treatments. Intracellular pH varied significantly (p < 0.01) in relation to either the incubation time or treatments. In particular, it showed a progressive increase throughout incubation with values in control group significantly higher than in myo-inositol, PHE, caffeine, pentoxifylline and CZA groups (7.37 ± 0.03 vs. 7.29 ± 0.03, 7.28 ± 0.03, 7.26 ± 0.03, 7.22 ± 0.03 and 7.00 ± 0.03, respectively; p < 0.01).; however, among treatments, CZA displayed the lowest values. Significant correlations were found between sperm kinetic and metabolic

  6. Intracellular pH regulation in isolated trout gill mitochondrion-rich (MR) cell subtypes: evidence for Na+/H+ activity.

    PubMed

    Parks, Scott K; Tresguerres, Martin; Galvez, Fernando; Goss, Greg G

    2010-02-01

    We have studied intracellular pH (pH(i)) recovery in isolated trout gill mitochondrion-rich (MR) cells following acidification by the NH(4)Cl pre-pulse technique. Within a mixed MR cell population, one cell type displayed Na(+)-independent pH(i) recovery while the other cell type lacked a Na(+)-independent pH(i) recovery. Cells displaying Na(+) independent recovery exhibited a significantly higher buffering capacity compared to cells lacking Na(+)-independent pH(i) recovery. Cells displaying Na(+) independent recovery were identified as PNA(+) (peanut lectin agluttinin binding) MR cells while those unable to recover were identified as PNA(-) (non-peanut lectin agluttinin binding) MR cells. Therefore, recovery from acidification in the absence of Na(+) provides a direct functional marker for PNA(+) and PNA(-) MR cells. Re-addition of Na(+) to acidified cells resulted in a transient pH(i) recovery in both cell types. This event was abolished by amiloride (500 microM) but it was insensitive to phenamil (50 microM). The phorbol ester PMA (1 microM) potentiated the Na(+) induced pH(i) recovery suggesting that activation by PKC is required for continuous Na(+)/H(+) exchanger activity in trout gill MR cells. This study is the first functional description of pH(i) recovery in lectin-identified trout gill MR cells and provides insight into a putative cellular signaling mechanism that may control pH(i) regulation in the gill epithelium.

  7. Effect of parsley (Petroselinum crispum (Mill.) Nym. ex A.W. Hill, Apiaceae) extracts on some biochemical parameters of oxidative stress in mice treated with CCl(4).

    PubMed

    Popović, Mira; Kaurinović, Biljana; Jakovljević, Vida; Mimica-Dukic, Neda; Bursać, Marija

    2007-08-01

    The in vitro and in vivo antioxidant activity of different extracts of leaves and root of parsley (Petroselinum crispum (Mill.) Nym. ex A.W. Hill, Apiaceae) were studied. Free radical scavenging capacity (RSC) was evaluated measuring the scavenging activity on the 2,2-diphenyl-1-picrylhydrazil (DPPH) and OH radicals. Also, the effects on lipid peroxidation (LP) were evaluated. The results obtained showed that all examined extracts act as good scavengers of DPPH and OH radicals and reduce the intensity of LP. The in vivo effects were evaluated on some antioxidant systems (activities of LPx, GSH-Px, Px, CAT and XOD, and GSH content) in the mice liver and blood after treatment with the examined parsley extracts, or in combination with carbon tetrachloride (CCl(4)). On the basis of the results obtained it can be concluded that the examined extracts exhibited a certain protective effect. However, combined treatments with CCl(4) and the examined extracts showed both positive and negative synergism, inducing or suppressing the influence of CCl(4) alone.

  8. Researchers Identify Genomic Alterations Associated with Drug-Targetable Kinase Activation in Ph-like Acute Lymphoblastic Leukemia | Office of Cancer Genomics

    Cancer.gov

    Acute lymphoblastic leukemia (ALL) is the most prevalent cancer among children and young adults, and standard treatments within this population generally result in favorable outcomes. By contrast, one particular subtype of this disease, Philadelphia chromosome-like ALL (Ph-like ALL), is associated with inferior outcomes. Ph-like ALL exhibits a gene expression profile similar to chromosome 9:22 translocation positive ALL, yet it lacks the characteristic BCR-ABL fusion protein.

  9. Boildown Study on Supernatant Liquid Retrieved from AW-106 in December 2012

    SciTech Connect

    Page, Jason S.

    2013-06-04

    This document reports the results of a boil down study using a composite created from supernatant liquid grab samples retrieved from tank 241-AW-I06 in December of 2012. The composite was made using predetermined volumes of the grab samples which accounted for layering of the supernatant liquid in the tank. The finished composite was a clear, yellow liquid containing no visible solids at hot cell ambient temperatures (24 - 27°C). The density of the test composite was measured in the hot cell immediately before the boildown study and was 1.266 g/mL at 27.1 °C. The boiling temperature of the composite was measured at three different pressures (40, 60, and 80 Torr) throughout the volume reduction, and the results show steadily increasing boiling temperatures with increasing volume reduction and no significant discontinuities. Moderate foaming was observed at the onset of the boildown. The foaming disappeared during the first reduction step, and minimal foaming was observed throughout the rest of the study. The bulk densities at 18.0 °C (D{sub Bulk}{sup 18 °C}) and quantities of settled and centrifuged solids were measured on samples of the boildown concentrates. Estimated values of the bulk densities at the 60-Torr boiling temperatures (D{sub Bulk}{sup 60 Torr}) were also calculated. Solids were first observed at boildown temperatures when the % VWR reached 39.3%. The quantity of solids in the composite quickly increased after this initial formation; the amount of centrifuged solids increased by 22% as the %WVR increased from 39.3 to 44.1 %. A small amount of solids did appear in the samples collected prior to the initial formation during the boildown. These solids precipitated while they sat at hot cell ambient temperature and in the 18. 0 °C water bath. Analysis of boil down test samples indicated that natrophosphate (Na7{sub 3}F(PO{sub 4}){sub 2}{centerdot} 19 H{sub 2}O) and kogarkoite (Na3FS04) accounted for a majority of the initial solids (~80% of the

  10. The dimeric assembly of Photobacterium leiognathi and Salmonella typhimurium SodC1 Cu,Zn superoxide dismutases is affected differently by active site demetallation and pH: an analytical ultracentrifuge study.

    PubMed

    Catacchio, B; D'Orazio, M; Battistoni, A; Chiancone, E

    2008-03-01

    To establish whether the species-specific variations at the subunit interface of bacterial Cu,Zn superoxide dismutases affect dimer assembly, the association state of the Photobacterium leiognathi (PlSOD) and Salmonella typhimurium (StSOD) enzymes, which differ in 11 out of 19 interface residues, was investigated by analytical ultracentrifugation. The same linkage pattern correlates quaternary assembly, active site metallation, and pH in the two enzymes albeit with quantitative differences. Both holo-enzymes are stable dimers at pH 6.8 and 8.0, although their shape is altered at alkaline pH. In contrast, dimer stability is affected differently by metal removal. Thus, apo-StSOD is a stable dimer at pH 6.8 whereas apo-PlSOD is in reversible monomer-dimer equilibrium. In both apoproteins a pH increase to 8.0 favors monomerization. These effects prove the existence of long-range communication between the active site and the subunit interface and provide a structural explanation for the known functional differences between the two enzymes.

  11. Breath gas metabolites and bacterial metagenomes from cystic fibrosis airways indicate active pH neutral 2,3-butanedione fermentation.

    PubMed

    Whiteson, Katrine L; Meinardi, Simone; Lim, Yan Wei; Schmieder, Robert; Maughan, Heather; Quinn, Robert; Blake, Donald R; Conrad, Douglas; Rohwer, Forest

    2014-06-01

    The airways of cystic fibrosis (CF) patients are chronically colonized by patient-specific polymicrobial communities. The conditions and nutrients available in CF lungs affect the physiology and composition of the colonizing microbes. Recent work in bioreactors has shown that the fermentation product 2,3-butanediol mediates cross-feeding between some fermenting bacteria and Pseudomonas aeruginosa, and that this mechanism increases bacterial current production. To examine bacterial fermentation in the respiratory tract, breath gas metabolites were measured and several metagenomes were sequenced from CF and non-CF volunteers. 2,3-butanedione was produced in nearly all respiratory tracts. Elevated levels in one patient decreased during antibiotic treatment, and breath concentrations varied between CF patients at the same time point. Some patients had high enough levels of 2,3-butanedione to irreversibly damage lung tissue. Antibiotic therapy likely dictates the activities of 2,3-butanedione-producing microbes, which suggests a need for further study with larger sample size. Sputum microbiomes were dominated by P. aeruginosa, Streptococcus spp. and Rothia mucilaginosa, and revealed the potential for 2,3-butanedione biosynthesis. Genes encoding 2,3-butanedione biosynthesis were disproportionately abundant in Streptococcus spp, whereas genes for consumption of butanedione pathway products were encoded by P. aeruginosa and R. mucilaginosa. We propose a model where low oxygen conditions in CF lung lead to fermentation and a decrease in pH, triggering 2,3-butanedione fermentation to avoid lethal acidification. We hypothesize that this may also increase phenazine production by P. aeruginosa, increasing reactive oxygen species and providing additional electron acceptors to CF microbes.

  12. The dependence on temperature and pH of the effects of zinc and copper on proteolytic activities of the digestive tract mucosa in piscivorous fish and their potential preys.

    PubMed

    Kuz'mina, V V; Ushakova, N V

    2010-09-01

    The dependence of the effects of zinc and copper on the activities of proteinases of the stomach and intestinal mucosa on temperature and pH in four species of boreal piscivorous fish (pike Esox lucius, zander Zander lucioperca, perch Perca fluviatilis and burbot Lota lota) as well as in some of their potential preys (kilka Clupeonella cultriventris, ruff Gymnocephalus cernuus, perch and roach Rutilus rutilus) was investigated. Species-specific differences of the effects of these heavy metals upon the activities of proteinases depending on temperature and pH were demonstrated. It was revealed that the stomach mucosa proteinases were more tolerant to the effects of the studied factors than the intestinal mucosa proteinases, especially true for pike. The effects of the heavy metals on the whole body proteinases of the fishes' potential preys were mostly dependent on temperature than on pH. At pH 3.0, the negative action of zinc and copper on the fish digestive tract mucosa proteolytic activity to a considerable degree was compensated by the high activity of the hemoglobinlytic proteinases, probably, cathepsine D.

  13. Impact of different pH control agents on biopesticidal activity of Bacillus thuringiensis during the fermentation of starch industry wastewater.

    PubMed

    Vu, Khanh Dang; Tyagi, R D; Valéro, J R; Surampalli, R Y

    2009-06-01

    Different pH control agents (NaOH/H(2)SO(4)--SodSulp, NaOH/CH(3)COOH--SodAcet, NH(4)OH/CH(3)COOH--AmmoAcet and NH(4)OH/H(2)SO(4)--AmmoSulp) were used to investigate their effects on growth, enzyme production (alkaline protease and amylase), and entomotoxicity of Bacillus thuringiensis var. kurstaki HD-1 (Btk) against eastern spruce budworm larvae (Choristoneura fumiferana) using starch industry wastewater (SIW) as a raw material in a 15-l fermentor. AmmoSulp and SodSulp were found to be the best pH control agents for alkaline protease and amylase production, respectively; whereas, the fermented broth obtained by using SodAcet as pH control agents recorded the highest delta-endotoxin production of 1043.0 mg/l and entomotoxicity value 18.4 x 10(9) SBU/l. Entomotoxicity of re-suspended centrifuged pellet in one-tenth of original volume in case of SodAcet as pH control agents was 26.7 x 10(9) SBU/l and was the highest value compared to three other pH control agents.

  14. Solute Transport Proteins and the Outer Membrane Protein NmpC Contribute to Heat Resistance of Escherichia coli AW1.7▿

    PubMed Central

    Ruan, Lifang; Pleitner, Aaron; Gänzle, Michael G.; McMullen, Lynn M.

    2011-01-01

    This study aimed to elucidate determinants of heat resistance in Escherichia coli by comparing the composition of membrane lipids, as well as gene expression, in heat-resistant E. coli AW1.7 and heat-sensitive E. coli GGG10 with or without heat shock. The survival of E. coli AW1.7 at late exponential phase was 100-fold higher than that of E. coli GGG10 after incubation at 60°C for 15 min. The cytoplasmic membrane of E. coli AW1.7 contained a higher proportion of saturated and cyclopropane fatty acids than that of E. coli GGG10. Microarray hybridization of cDNA libraries obtained from exponentially growing or heat-shocked cultures was performed to compare gene expression in these two strains. Expression of selected genes from different functional groups was quantified by quantitative PCR. DnaK and 30S and 50S ribosomal subunits were overexpressed in E. coli GGG10 relative to E. coli AW1.7 upon heat shock at 50°C, indicating improved ribosome stability. The outer membrane porin NmpC and several transport proteins were overexpressed in exponentially growing E. coli AW1.7. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of membrane properties confirmed that NmpC is present in the outer membrane of E. coli AW1.7 but not in that of E. coli GGG10. Expression of NmpC in E. coli GGG10 increased survival at 60°C 50- to 1,000-fold. In conclusion, the outer membrane porin NmpC contributes to heat resistance in E. coli AW1.7, but the heat resistance of this strain is dependent on additional factors, which likely include the composition of membrane lipids, as well as solute transport proteins. PMID:21398480

  15. Development of a model describing the effect of temperature, water activity and (gel) structure on growth and ochratoxin A production by Aspergillus carbonarius in vitro and evaluation in food matrices of different viscosity.

    PubMed

    Kapetanakou, Anastasia E; Ampavi, Anna; Yanniotis, Stavrianos; Drosinos, Eleftherios H; Skandamis, Panagiotis N

    2011-06-01

    The present study aimed: (i) to develop models for the combined effect of water activity (0.99, 0.94 and 0.90), microstructure expressed as 0, 5, 10 and 20% w/v gelatin, and temperature (15, 20 and 25 °C), on growth and OTA production rates by Aspergillus carbonarius; and (ii) to evaluate the performance of the developed models on food matrices (jelly, custard and marmalade) of different viscosity at pH 5.5. The square root of biomass increase rate (fungal growth rate) and OTA production rate were determined by the Baranyi model and were further modeled as a function of temperature, gelatin concentration and a(w) by applying polynomial models. Time for visible growth and the upper asymptote of the OTA production curve were also determined by the Baranyi model. Increase in gelatin concentration resulted in a significant delay in all parameters describing fungal growth and OTA production rates, at all temperatures and a(w). The effect of microstructure on fungal growth and OTA production rates was less evident at stress conditions of a(w) and temperature. Detection time for visible fungal growth was markedly influenced by a(w) and temperature. Coefficients of determination were 0.899 and 0.887 for the models predicting the square root (√μ(max)) of growth and OTA production rate, respectively. Predictions of growth rate agreed well with the recorded data of custard and marmalade, while observations of OTA production rate indicated low agreement with model predictions, in all food matrices except for marmalade. The present findings may provide a basis for reliable assessment of the risk of fungal growth and OTA production in foods of different structural and rheological properties.

  16. Evaluation of Flow Rate, pH, Buffering Capacity, Calcium, Total Proteins and Total Antioxidant Capacity Levels of Saliva in Caries Free and Caries Active Children: An In Vivo Study.

    PubMed

    Preethi, B P; Reshma, Dodawad; Anand, Pyati

    2010-10-01

    The purpose of this study was to evaluate the relationship between the physicochemical properties of saliva such as flow rate, pH, buffering capacity, calcium level, total protein and total antioxidant levels in caries free and caries active children. The present study included one hundred and twenty healthy children who were divided into two groups; group I and group II comprising of age groups 7-10 and 11-14 years, respectively. Both the groups were then sub-divided equally according to gender. They were further divided into caries free and caries active with 15 children in each group. Unstimulated saliva was collected by suction method and flow rates were determined. The samples were then analyzed for pH, buffering capacity, total protein, calcium and total antioxidant capacity. The data was statistically analyzed using student t test (unpaired). The results revealed that when all these parameters were compared among the caries free and caries active children, flow rate, pH, buffering capacity were slightly reduced in caries active children, but total protein and total antioxidant capacity of saliva increased significantly in caries active children and the total calcium decreased significantly in caries active children. Within the limitation of this study, we conclude that, the physicochemical properties of saliva play a major role in the development of caries.

  17. Internal Active Thermal Control System (IATCS) Sodium Bicarbonate/Carbonate Buffer in an Open Aqueous Carbon Dioxide System and Corollary Electrochemical/Chemical Reactions Relative to System pH Changes

    NASA Technical Reports Server (NTRS)

    Stegman, Thomas W.; Wilson, Mark E.; Glasscock, Brad; Holt, Mike

    2014-01-01

    The International Space Station (ISS) Internal Active Thermal Control System (IATCS) experienced a number of chemical changes driven by system absorption of CO2 which altered the coolant’s pH. The natural effects of the decrease in pH from approximately 9.2 to less than 8.4 had immediate consequences on system corrosion rates and corrosion product interactions with specified coolant constituents. The alkalinity of the system was increased through the development and implementation of a carbonate/bicarbonate buffer that would increase coolant pH to 9.0 – 10.0 and maintain pH above 9.0 in the presence of ISS cabin concentrations of CO2 up to twenty times higher than ground concentrations. This paper defines how a carbonate/bicarbonate buffer works in an open carbon dioxide system and summarizes the analyses performed on the buffer for safe and effective application in the on-orbit system. The importance of the relationship between the cabin environment and the IATCS is demonstrated as the dominant factor in understanding the system chemistry and pH trends before and after addition of the carbonate/bicarbonate buffer. The paper also documents the corollary electrochemical and chemical reactions the system has experienced and the rationale for remediation of these effects with the addition of the carbonate/bicarbonate buffer.

  18. Modeling the effect of water activity and storage temperature on chemical stability of coffee brews.

    PubMed

    Manzocco, Lara; Nicoli, Maria Cristina

    2007-08-08

    This work was addressed to study the chemical stability of coffee brew derivatives as a function of water activity (aw) and storage temperature. To this purpose, coffee brew was freeze-dried, equilibrated at increasing aw values, and stored for up to 10 months at different temperatures from -30 to 60 degrees C. The chemical stability of the samples was assessed by measuring H3O+ formation during storage. Independently of storage temperature, the rate of H3O+ formation was considerably low only when aw was reduced below 0.5 (94% w/w). Beyond this critical boundary, the rate increased, reaching a maximum value at ca. 0.8 aw (78% w/w). Further hydration up to the aw of the freshly prepared beverage significantly increased chemical stability. It was suggested that mechanisms other than lactones' hydrolysis, probably related to nonenzymatic browning pathways, could contribute to the observed increase in acidity during coffee staling. The temperature dependence of H3O+ formation was well-described by the Arrhenius equation in the entire aw range considered. However, aw affected the apparent activation energy and frequency factor. These effects were described by simple equations that were used to set up a modified Arrhenius equation. This model was validated by comparing experimental values, not used to generate the model, with those estimated by the model itself. The model allowed efficient prediction of the chemical stability of coffee derivatives on the basis of only the aw value and storage temperature.

  19. pH [Measure of Acidity].

    ERIC Educational Resources Information Center

    Henderson, Paula

    This autoinstructional program deals with the study of the pH of given substances by using litmus and hydrion papers. It is a learning activity directed toward low achievers involved in the study of biology at the secondary school level. The time suggested for the unit is 25-30 minutes (plus additional time for further pH testing). The equipment…

  20. Novel pH control strategy for efficient production of optically active l-lactic acid from kitchen refuse using a mixed culture system.

    PubMed

    Tashiro, Yukihiro; Inokuchi, Shota; Poudel, Pramod; Okugawa, Yuki; Miyamoto, Hirokuni; Miayamoto, Hisashi; Sakai, Kenji

    2016-09-01

    Uninvestigated control factors of meta-fermentation, the fermentative production of pure chemicals and fuels in a mixed culture system, were examined for production of optically pure l-lactic acid (LA) from food waste. In meta-fermentations by pH swing control, l-LA production with 100% optical purity (OPl-LA) was achieved even using unsterilized model kitchen refuse medium with preferential proliferation of l-LA-producing Bacillus coagulans, a minor member in the seed, whereas agitation decreased OPl-LA drastically. pH constant control shortened the fermentation time but decreased OPl-LA and LA selectivity (SLA) by stimulating growth of heterofermentative Bacillus thermoamylovorans. Deliberately switching from pH swing control to constant control exhibited the best performance for l-LA production: maximum accumulation, 39.2gL(-1); OPl-LA, 100%; SLA, 96.6%; productivity, 1.09gL(-1)h(-1). These results present a novel pH control strategy for efficient l-LA production in meta-fermentation based on a concept different from that of pure culture systems.

  1. Electrostatic Assemblies of Well-Dispersed AgNPs on the Surface of Electrospun Nanofibers as Highly Active SERS Substrates for Wide-Range pH Sensing.

    PubMed

    Yang, Tong; Ma, Jun; Zhen, Shu Jun; Huang, Cheng Zhi

    2016-06-15

    Surface-enhanced Raman scattering (SERS) has shown high promise in analysis and bioanalysis, wherein noble metal nanoparticles (NMNPs) such as silver nanoparticles were employed as substrates because of their strong localized surface plasmon resonance (LSPR) properties. However, SERS-based pH sensing was restricted because of the aggregation of NMNPs in acidic medium or biosamples with high ionic strength. Herein, by using the electrostatic interaction as a driving force, AgNPs are assembled on the surface of ethylene imine polymer (PEI)/poly(vinyl alcohol) (PVA) electrospun nanofibers, which are then applied as highly sensitive and reproducible SERS substrate with an enhancement factor (EF) of 10(7)-10(8). When p-aminothiophenol (p-ATP) is used as an indicator with its b2 mode, a good and wide linear response to pH ranging from 2.56 to 11.20 could be available, and the as-prepared nanocomposite fibers then could be fabricated as excellent pH sensors in complicated biological samples such as urine, considering that the pH of urine could reflect the acid-base status of a person. This work not only emerges a cost-effective, direct, and convenient approach to homogeneously decorate AgNPs on the surface of polymer nanofibers but also supplies a route for preparing other noble metal nanofibrous sensing membranes.

  2. Au@Ag Heterogeneous Nanorods as Nanozyme Interfaces with Peroxidase-Like Activity and Their Application for One-Pot Analysis of Glucose at Nearly Neutral pH.

    PubMed

    Han, Lei; Li, Cuncheng; Zhang, Tao; Lang, Qiaolin; Liu, Aihua

    2015-07-08

    As substitutes for natural peroxidases, most nanomaterial-based enzyme mimetics (nanozymes) have unique properties such as high stability, low-cost, large surface area, and high catalytic activity. However, they usually work in acidic conditions and thus impede their real applications. In this work, by modulating the nanostructure, composition, and surface property of the bimetallic materials, the positively charged poly(diallyldimethylammonium)-stabilized Au@Ag heterogeneous nanorods (NRs) were developed as synergistic peroxidase-like interfaces, which exhibited high activity over a wide pH range (pH 4.0-6.5) using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) as the chromogenic substrate. At pH 6.5, the peroxidase-like activity for the Au@Ag heterogeneous NRs was stable and optimal within 20-40 °C. Moreover, the Au@Ag heterogeneous NRs showed excellent temperature stability and long-term storage stability. Given these characters, the detection of H2O2 at pH 6.5 was proposed on the basis of the Au@Ag heterogeneous NRs catalyzing the colorimetric reaction of H2O2 and ABTS, where the oxidized ABTS showed a typical absorption peak at 414 nm. The absorbance at 414 nm was linear with H2O2 concentration from 0.01 to 10 mM. Further, considering that Au@Ag heterogeneous NRs and glucose oxidase (GOx) have similar optimal pH for catalytic activities, a novel one-pot method for the detection of glucose was developed by the coupled catalytic reaction using GOx, Au@Ag heterogeneous NRs, and ABTS at nearly neutral pH (pH 6.5) and 37 °C. This proposed method had simple and rapid processes, wide linear range (0.05-20 mM), and reliability for the successful analysis of real samples. On the basis of these attractive and unique characteristics, Au@Ag heterogeneous NRs can become promising substitutes for peroxidase in analytical chemistry and environmental science.

  3. Functional photoacoustic microscopy of pH

    NASA Astrophysics Data System (ADS)

    Chatni, M. Rameez; Yao, Junjie; Danielli, Amos; Favazza, Christopher P.; Maslov, Konstantin I.; Wang, Lihong V.

    2012-02-01

    pH is a tightly regulated indicator of metabolic activity. In mammalian systems, imbalance of pH regulation may result from or result in serious illness. Even though the regulation system of pH is very robust, tissue pH can be altered in many diseases such as cancer, osteoporosis and diabetes mellitus. Traditional high-resolution optical imaging techniques, such as confocal microscopy, routinely image pH in cells and tissues using pH sensitive fluorescent dyes, which change their fluorescence properties with the surrounding pH. Since strong optical scattering in biological tissue blurs images at greater depths, high-resolution pH imaging is limited to penetration depths of 1mm. Here, we report photoacoustic microscopy (PAM) of commercially available pH-sensitive fluorescent dye in tissue phantoms. Using both opticalresolution photoacoustic microscopy (OR-PAM), and acoustic resolution photoacoustic microscopy (AR-PAM), we explored the possibility of recovering the pH values in tissue phantoms. In this paper, we demonstrate that PAM was capable of recovering pH values up to a depth of 2 mm, greater than possible with other forms of optical microscopy.

  4. Dandelion PPO-1/PPO-2 domain-swaps: the C-terminal domain modulates the pH optimum and the linker affects SDS-mediated activation and stability.

    PubMed

    Leufken, Christine M; Moerschbacher, Bruno M; Dirks-Hofmeister, Mareike E

    2015-02-01

    Plant polyphenol oxidases (PPOs) have a conserved three-domain structure: (i) the N-terminal domain (containing the active site) is connected via (ii) a linker to (iii) the C-terminal domain. The latter covers the active site, thereby maintaining the enzyme in a latent state. Activation can be achieved with SDS but little is known about the mechanism. We prepared domain-swap variants of dandelion PPO-1 and PPO-2 to test the specific functions of individual domains and their impact on enzyme characteristics. Our experiments revealed that the C-terminal domain modulates the pH optimum curve and has a strong influence on the optimal pH value. The linker determines the SDS concentration required for full activation. It also influences the SDS concentration required for half maximal activation (kSDS) and the stability of the enzyme during prolonged incubation in buffers containing SDS, but the N-terminal domain has the strongest effect on these parameters. The N-terminal domain also determines the IC50 of SDS and the stability in buffers containing or lacking SDS. We propose that the linker and C-terminal domain fine-tune the activation of plant PPOs. The C-terminal domain adjusts the pH optimum and the linker probably contains an SDS-binding/interaction site that influences inactivation and determines the SDS concentration required for activation. For the first time, we have determined the influence of the three PPO domains on enzyme activation and stability providing insight into the regulation and activation mechanisms of type-3 copper proteins in general.

  5. Report on Foundation and Organization of Econophysics Graduate Courses at Faculty of Physics of University of Warsaw and Department of Physics and Astronomy of the Wrocław University

    NASA Astrophysics Data System (ADS)

    Kutner, R.; Grech, D.

    2008-09-01

    Two different, working examples of organization of econophysics graduate courses at the Faculty of Physics, University of Warsaw and the Department of Physics and Astronomy of the Wrocław University are considered. In the first example we have a system where the interdisciplinary, econophysical education begins only after three years study of physics. Within this system the M.Sc. as well as Ph.D. theses in econophysics are conducted only at the Faculty of Physics. In the second example the B.Sc. theses in econophysics are accomplished in the Department of Physics and Astronomy again after three years study but higher degrees can be prepared either in physics in the Institute of Theoretical Physics or in economy in the Institute of Economical Sciences. M.Sc. and Ph.D. theses can also be conducted. For both examples, the graduate students of econophysics are obliged to participate in traditional (typical) economical lectures and trainings which are offered them by economical departments while lectures and trainings (tutorials and/or laboratory classes) in econophysics are offered them by physics departments themselves. Thus Poland is one of a few countries, where so modern interdisciplinary knowledge is systematically offered to students.

  6. A study on the physicochemical parameters for Penicillium expansum growth and patulin production: effect of temperature, pH, and water activity.

    PubMed

    Tannous, Joanna; Atoui, Ali; El Khoury, André; Francis, Ziad; Oswald, Isabelle P; Puel, Olivier; Lteif, Roger

    2016-07-01

    Penicillium expansum is among the most ubiquitous fungi disseminated worldwide, that could threaten the fruit sector by secreting patulin, a toxic secondary metabolite. Nevertheless, we lack sufficient data regarding the growth and the toxigenesis conditions of this species. This work enables a clear differentiation between the favorable conditions to the P. expansum growth and those promising for patulin production. A mathematical model allowing the estimation of the P. expansum growth rate according to temperature, a W, and pH, was also developed. An optimal growth rate of 0.92 cm/day was predicted at 24°C with pH level of 5.1 and high a W level of 0.99. The model's predictive capability was tested successfully on artificial contaminated apples. This model could be exploited by apple growers and the industrialists of fruit juices in order to predict the development of P. expansum during storage and apple processing.

  7. pH control strategy in a shaken minibioreactor for polysaccharide production by medicinal mushroom Phellinus linteus and its anti-hyperlipemia activity.

    PubMed

    Zou, Xiang; Guo, Xia; Sun, Min

    2009-02-01

    A process at various pH values ranging from 4.5 to 7.5 for production of mycelia and extracellular polysaccharide (EPS) by P. linteus fermentation in pH-controlled shaken bioreactor was investigated. A two-stage pH control strategy in which pH value was kept at 6.5 for the first 24 h, and then switched to 4.5 was developed successfully to enhance simultaneously the cell growth and EPS production. The maximum cell density and EPS production reached 15.13 +/- 0.1 g/l on day 6 and 6.74 +/- 0.1 g/l on day 4, respectively. The anti- hyperlipemia effect of EPS and intracellular polysaccharide (IPS) extracted from mycelia were observed that both EPS and IPS can obviously reduce the serum triglyceride (TG), the blood cholesterol (TC) and serum low density lipoprotein (LDL) level, and increase the high density lipoprotein (HDL) level of the hyperlipemia mice. Polysaccharides from submerged cultivation of medicinal fungus P. linteus have favorable potency to develop anti-hyperlipermia drugs.

  8. Relative survival of four serotypes of Salmonella enterica in low-water activity whey protein powder held at 36 and 70°C at various water activity levels.

    PubMed

    Farakos, S M Santillana; Hicks, J W; Frye, J G; Frank, J F

    2014-07-01

    Salmonella enterica is not able to grow at water activity (aw) levels below 0.94, but it can survive in low-aw foods for long periods of time. Temperature, aw, substrate, and serotype affect its persistence. The aim of this study was to evaluate the influence of temperature and aw on the relative persistence among four serotypes of Salmonella enterica in low-aw whey protein powder. Whey protein powder was equilibrated to aws 0.18 ± 0.02 and 0.54 ± 0.03, inoculated with a cocktail of Salmonella serovars (Agona, Tennessee, Montevideo, and Typhimurium), vacuum sealed, and stored at 36°C for 6 months and at 70°C for 48 h. Presumptive Salmonella colonies (30 to 32) were randomly picked from each plate at the end of each survival study. PCR multiplex serotyping was used to identify the isolates. A multinomial mixed logistic model with Salmonella Tennessee as a reference was used to test for significant differences in frequency distribution of the surviving serotypes. Salmonella Tennessee and Salmonella Agona were the most prevalent surviving serotypes, followed in decreasing order by Salmonella Montevideo and Salmonella Typhimurium. Statistical analysis indicated that temperature (P = 0.003) and aw (P = 0.012) influenced the relative prevalence of the Salmonella serotypes. If other environmental conditions are equal, Salmonella Tennessee is better able to survive than Salmonella Montevideo and Salmonella Typhimurium at higher temperatures and higher aw levels in low-aw whey protein powder held at 36 and 70°C. The relative prevalence of Salmonella Agona to Salmonella Tennessee did not change with increasing temperature (P = 0.211) or aw (P = 0.453). These results should be considered in risk assessment and when developing predictive models for survival of Salmonella in low-aw foods.

  9. The activity of TcCYS4 modified by variations in pH and temperature can affect symptoms of witches' broom disease of cocoa, caused by the fungus Moniliophthora perniciosa.

    PubMed

    Freitas, Ana Camila Oliveira; Souza, Cristiane Ferreira; Monzani, Paulo Sérgio; Garcia, Wanius; de Almeida, Alex Alan Furtado; Costa, Marcio Gilberto Cardoso; Pirovani, Carlos Priminho

    2015-01-01

    The phytocystatins regulate various physiological processes in plants, including responses to biotic and abiotic stresses, mainly because they act as inhibitors of cysteine proteases. In this study, we have analyzed four cystatins from Theobroma cacao L. previously identified in ESTs libraries of the interaction with the fungus Moniliophthora perniciosa and named TcCYS1, TcCYS2, TcCYS3 and TcCYS4. The recombinant cystatins were purified and subjected to the heat treatment, at different temperatures, and their thermostabilities were monitored using their ability to inhibit papain protease. TcCYS1 was sensitive to temperatures above 50°C, while TcCYS2, TcCYS3, and TcCYS4 were thermostable. TcCYS4 presented a decrease of inhibitory activity when it was treated at temperatures between 60 and 70°C, with the greater decrease occurring at 65°C. Analyses by native gel electrophoresis and size-exclusion chromatography showed that TcCYS4 forms oligomers at temperatures between 60 and 70°C, condition where reduction of inhibitory activity was observed. TcCYS4 oligomers remain stable for up to 20 days after heat treatment and are undone after treatment at 80°C. TcCYS4 presented approximately 90% of inhibitory activity at pH values between 5 and 9. This protein treated at temperatures above 45°C and pH 5 presented reduced inhibitory activity against papain, suggesting that the pH 5 enhances the formation of TcCYS4 oligomers. A variation in the titratable acidity was observed in tissues of T. cacao during the symptoms of witches' broom disease. Our findings suggest that the oligomerization of TcCYS4, favored by variations in pH, is an endergonic process. We speculate that this process can be involved in the development of the symptoms of witches' broom disease in cocoa.

  10. Arecibo and Goldstone Radar Observations of the First-Recognized Binary Near-Earth Asteroid: (385186) 1994 AW1

    NASA Astrophysics Data System (ADS)

    Richardson, James E.; Taylor, Patrick A.; Rivera-Valentin, Edgard G.; Rodriguez-Ford, Linda A.; Benner, Lance A. M.; Naidu, Shantanu P.; Giorgini, Jon D.; Busch, Michael W.; Ghigo, Frank d.; Kobelski, Adam; Warner, Brian D.; Springmann, Alessondra; Marshall, Sean E.; Steckloff, Jordan K.; Sharkey, Benjamin

    2015-11-01

    Near-Earth asteroid (385186) 1994 AW1 was discovered at Palomar Observatory on 11 January 1994. Subsequent observations of this Amor family, Sa-class asteroid also identified it as the first candidate binary NEA, as indicated by multiple periodicities and possible mutual eclipsing/occulting events in the object’s lightcurve. On 15 July 2015 this asteroid made its closest approach to Earth since its discovery, coming within 0.065 AU (25 lunar distances), and prompting an extended observation campaign using both the JPL-Goldstone and Arecibo Observatory planetary radars. Goldstone observations covered the 14-19 July period of closest approach (0.066-0.070 AU) while the object remained below Arecibo’s observing horizon, with Arecibo picking up the observations between 20-30 July, as the object moved from 0.075 to 0.126 AU distance. At Goldstone, we were able to observe this object with range resolutions of 150 m using a Goldstone (DSS-14) to Green Bank Telescope (GBT) bistatic configuration, while at Arecibo, we conducted monostatic observations of 1994 AW1 using the 2380 MHz (12.6 cm) radar at resolutions of 30 m and 75 m.As a result, and twenty years after its discovery, these observations have confirmed the binary nature of 1994 AW1, showing the primary body to be about 600 m in diameter, the secondary body to be about half the diameter of the primary, with the two orbiting a common center of mass at a distance of about 1.2 km apart. Delay-Doppler image comparisons of the primary over the course of six nights (at 30 m resolution) confirm a lightcurve-derived rotation period of 2.518 +/- 0.002 hr, as >90% longitude coverage was achieved, revealing a slightly elongated, irregular surface morphology. Delay-Doppler images of the secondary reveal an elongated, irregular body which appears to be tidally locked, with its long axis pointed towards the primary as it orbits with a period of about 22 hr (also consistent with the lightcurve analysis). These very early

  11. Time sequence spectroscopy of AW UMa. The 518 nm Mg I triplet region analyzed with broadening functions

    SciTech Connect

    Rucinski, Slavek M.

    2015-02-01

    High-resolution spectroscopic observations of AW UMa, obtained on three consecutive nights with a median time resolution of 2.1 minutes, have been analyzed using the broadening function method in the spectral window of 22.75 nm around the 518 nm Mg i triplet region. Doppler images of the system reveal the presence of vigorous mass motions within the binary system; their presence puts into question the solid-body rotation assumption of the contact binary model. AW UMa appears to be a very tight, semi-detached binary; the mass transfer takes place from the more massive to the less massive component. The primary, a fast-rotating star with Vsini=181.4±2.5 km s{sup −1}, is covered with inhomogeneities: very slowly drifting spots and a dense network of ripples more closely participating in its rotation. The spectral lines of the primary show an additional broadening component (called the “pedestal”) that originates either in the equatorial regions, which rotate faster than the rest of the star by about 50 km s{sup −1}, or in an external disk-like structure. The secondary component appears to be smaller than predicted by the contact model. The radial velocity field around the secondary is dominated by accretion of matter transferred from (and possibly partly returned to) the primary component. The parameters of the binary are Asini=2.73±0.11 R{sub ⊙} and M{sub 1}sin{sup 3}i=1.29±0.15 M{sub ⊙}, M{sub 2}sin{sup 3}i=0.128±0.016 M{sub ⊙}. The mass ratio, q{sub sp}=M{sub 2}/M{sub 1}=0.099±0.003, while still the most uncertain among the spectroscopic elements, is substantially different from the previous numerous and mutually consistent photometric investigations which were based on the contact model. It should be studied why photometry and spectroscopy give such discrepant results and whether AW UMa is an unusual object or if only very high-quality spectroscopy can reveal the true nature of W UMa-type binaries.

  12. Acidic pH and short-chain fatty acids activate Na+ transport but differentially modulate expression of Na+/H+ exchanger isoforms 1, 2, and 3 in omasal epithelium.

    PubMed

    Lu, Zhongyan; Yao, Lei; Jiang, Zhengqian; Aschenbach, Jörg R; Martens, Holger; Shen, Zanming

    2016-01-01

    Low sodium content in feed and large amounts of salivary sodium secretion are essential requirements to efficient sodium reabsorption in the dairy cow. It is already known that Na(+)/H(+) exchange (NHE) of the ruminal epithelium plays a key role in Na(+) absorption, and its function is influenced by the presence of short-chain fatty acids (SCFA) and mucosal pH. By contrast, the functional role and regulation of NHE in omasal epithelium have not been completely understood. In the present study, we used model studies in small ruminants (sheep and goats) to investigate NHE-mediated Na(+) transport and the effects of pH and SCFA on NHE activity in omasal epithelium and on the expression of NHE isoform in omasal epithelial cells. Conventional Ussing chamber technique, primary cell culture, quantitative PCR, and Western blot were used. In native omasal epithelium of sheep, the Na(+) transport was electroneutral, and it was inhibited by the specific NHE3 inhibitor 3-[2-(3-guanidino-2-methyl-3-oxo-propenyl)-5-methyl-phenyl]-N-isopropylidene-2-methyl-acrylamide dihydrochloride, which decreased mucosal-to-serosal, serosal-to-mucosal, and net flux rates of Na(+) by 80% each. The application of low mucosal pH (6.4 or 5.8) in the presence of SCFA activated the Na(+) transport across omasal epithelium of sheep compared with that at pH 7.4. In cultured omasal epithelial cells of goats, mRNA and protein of NHE1, NHE2, and NHE3 were detected. The application of SCFA increased NHE1 mRNA and protein expression, which was most prominent when the culture medium pH decreased from 7.4 to 6.8. At variance, the mRNA and protein expression of NHE2 and NHE3 were decreased with low pH and SCFA, which was contrary to the published data from ruminal epithelial studies. In conclusion, this paper shows that (1) NHE1, NHE2, and NHE3 are expressed in omasal epithelium; (2) NHE3 mediates the major portion of transepithelial Na(+) transport in omasal epithelium; and (3) SCFA and acidic pH acutely

  13. Biological activity of neutral and cationic iridium(III) complexes with κP and κP,κS coordinated Ph₂PCH₂S(O)xPh (x = 0-2) ligands.

    PubMed

    Ludwig, Gerd; Mijatović, Sanja; Ranđelović, Ivan; Bulatović, Mirna; Miljković, Djordje; Maksimović-Ivanić, Danijela; Korb, Marcus; Lang, Heinrich; Steinborn, Dirk; Kaluđerović, Goran N

    2013-11-01

    Neutral iridium(III) complexes of the type [Ir(η(5)-C₅Me₅)Cl₂{Ph₂PCH₂S(O)xPh-κP}] (1-3) with diphenylphosphino-functionalized methyl phenyl sulfides, sulfoxides, and sulfones Ph₂PCH₂S(O)xPh (x = 0, L1; 1, L2; 2, L3) and the cationic complex [Ir(η(5)-C₅Me₅)Cl{Ph₂PCH₂SPh-κP,κS}][PF6] (4) were synthesized and fully characterized analytically and spectroscopically. Furthermore, the structure of 2 was determined by X-ray diffraction analysis. The biological potential of the neutral and cationic iridium(III) complexes was tested in vitro against the cell lines 8505C, A253, MCF-7, SW480 and 518A2. Complex [Ir(η(5)-C₅Me₅)Cl₂{Ph₂PCH₂S(O)Ph-κP}] (2), with ligand L2 κP coordinated containing a pendent sulfinyl group, is the most active one (IC₅₀ values of about 3 μM), thus, with activities comparable to cisplatin. Complex 2 proved to have an even a higher antiproliferative activity than cisplatin against 8505C and SW480 cell lines, used as a model system of highly anaplastic cancers with low sensitivity to conventional chemotherapeutics such as cisplatin. Additional experiments demonstrated that apoptosis and autophagic cell death contribute to the drug's tumoricidal action.

  14. Bis(alkyl) rare-earth complexes coordinated by bulky tridentate amidinate ligands bearing pendant Ph2P[double bond, length as m-dash]O and Ph2P[double bond, length as m-dash]NR groups. Synthesis, structures and catalytic activity in stereospecific isoprene polymerization.

    PubMed

    Rad'kova, Natalia Yu; Tolpygin, Aleksei O; Rad'kov, Vasily Yu; Khamaletdinova, Nadia M; Cherkasov, Anton V; Fukin, Georgi K; Trifonov, Alexander A

    2016-11-22

    A series of new tridentate amidines 2-[Ph2P[double bond, length as m-dash]X]C6H4NHC(tBu)[double bond, length as m-dash]N(2,6-R2C6H3) (X = O, R = iPr (1); X = S, R = Me (2); X = NPh, R = Me (3); X = N(2,6-Me2C6H3), R = Me (4)) bearing various types of donor Ph2P[double bond, length as m-dash]X groups in a pendant chain was synthesized. Bis(alkyl) complexes {2-[Ph2P[double bond, length as m-dash]X]C6H4NC(tBu)N(2,6-R2C6H3)}Ln(CH2SiMe3)2 (Ln = Y, X = O, R = iPr (5); Ln = Er, X = O, R = iPr (6); Ln = Lu, X = O, R = iPr (7); Ln = Y, X = NPh, R = Me (8); Ln = Lu, X = NPh, R = Me (9); Ln = Lu, X = N(2,6-Me2C6H3), R = Me (10)) were prepared using alkane elimination reactions of 1, 3 and 4 with Ln(CH2SiMe3)3(THF)2 (Ln = Y, Er, Lu) in toluene and were isolated in 45 (5), 62 (6), 56 (7), 65 (8), 60 (9), and 60 (10) % yields respectively. The X-ray diffraction studies showed that complexes 6-8 are solvent free and feature intramolecular coordination of the P[double bond, length as m-dash]X (X = O, NPh) group to the lanthanide ions. The ternary systems 5-10/borate/AlR3 (borate = [PhNHMe2][B(C6F5)4], [Ph3C][B(C6F5)4], AlR3 = AliBu3, AliBu2H; molar ratio = 1/1/10 or 1/1/1, toluene) proved to be active in isoprene polymerization and enable complete conversion of 1000-10 000 equivalents of the monomer into a polymer at 25 °C within 0.5-24 h affording polyisoprenes with polydispersities Mw/Mn = 1.22-3.18. A comparative study of the catalytic performance of the bis(alkyl) complexes coordinated by tridentate amidinate ligands containing different pendant donor groups demonstrated that replacement of the Ph2P[double bond, length as m-dash]O group by Ph2P[double bond, length as m-dash]NPh leads to a switch of stereoselectivity in isoprene polymerization from cis-1,4 (up to 98.5%) to trans-1,4 (up to 84.8%). And conversely introduction of methyl substituents in the 2,6 positions of the phenyl group of the Ph2P[double bond, length as m-dash]NPh fragment allows us to restore the 1,4-cis

  15. Comparisons of ANS, ASME, AWS, and NFPA standards cited in the NRC standard review plan, NUREG-0800, and related documents

    SciTech Connect

    Ankrum, A.R.; Bohlander, K.L.; Gilbert, E.R.; Spiesman, J.B.

    1995-11-01

    This report provides the results of comparisons of the cited and latest versions of ANS, ASME, AWS and NFPA standards cited in the NRC Standard Review Plan for the Review of Safety Analysis Reports for Nuclear Power Plants (NUREG 0800) and related documents. The comparisons were performed by Battelle Pacific Northwest Laboratories in support of the NRC`s Standard Review Plan Update and Development Program. Significant changes to the standards, from the cited version to the latest version, are described and discussed in a tabular format for each standard. Recommendations for updating each citation in the Standard Review Plan are presented. Technical considerations and suggested changes are included for related regulatory documents (i.e., Regulatory Guides and the Code of Federal Regulations) citing the standard. The results and recommendations presented in this document have not been subjected to NRC staff review.

  16. [Trends of faunistic-ecological researches in the Department of Parasitology in Wrocław University].

    PubMed

    Okulewicz, Anna; Lonc, Elzbieta

    2007-01-01

    Paper deals with the results of systematic-ecological investigations of host-parasite relationships in the context of 45 year history of the Parasitological Department at the University of Wrocław. Achievements are concentrated on the presentations of new taxonomic units and rearrangement of classification of parasitic protozoans and tapeworms (J. Janiszewska), nematodes (A. Okulewicz), lice (J. Złotorzycka, E. Lonc, M. Modrzejewska). Faunistic and biometrical data of parasitic commities of fishes, birds and mammals, mainly rodents as well as the vector role of ticks and mosquitoes at Wroclaw area are also discussed. Localities of those parasites and their hosts, collected mainly from the Lower Silesia region are rearranged according to new system of physicogeographic differentiation of Poland and its regional position at biogeographical map of world.

  17. Laser and pyrolysis removal of fluorinated ethylene propylene thin layers applied on EN AW-5251 aluminium substrates

    NASA Astrophysics Data System (ADS)

    Guerrero, G. R.; Sevilla, L.; Soriano, C.

    2015-10-01

    This work compares laser and pyrolysis removal procedures of the fluorinated ethylene propylene FEP-rich coatings applied on EN AW-5251 H34 aluminium magnesium alloy sheets. The study has been conducted in three successive FEP coating application-removal cycles. Roughness, yield and tensile strength, elongation percentage, hardness, grain size and constituent particles distribution of the processed samples have been analyzed. According to the results, it is concluded that similar mechanical properties has been obtained for both removal technologies, being the laser decoating removal rates lower than those achieved by the pyrolytic procedure. However, the laser decoating process is consolidated as an industrial alternative against the dangerous and harmful (to the environment) pyrolytic extraction systems.

  18. Staphylococcal nuclease active-site amino acids: pH dependence of tyrosines and arginines by sup 13 C NMR and correlation with kinetic studies

    SciTech Connect

    Grissom, C.G.; Markley, J.L. )

    1989-03-07

    The pH and temperature dependence of the kinetic parameters of staphylococcal nuclease have been examined with three p-nitrophenyl phosphate containing DNA analogues that vary as to 3'-substituent. With wild-type (Foggi variant) nuclease (nuclease wt) and the substrates thymidine 3'-phosphate 5'-(p-nitrophenyl phosphate) (PNPdTp), thymidine 3'-methylphosphonate 5'-(p-nitrophenyl phosphate) (PNPdTp Me), and thymidine 5'-(p-nitrophenyl phosphate) (PNPdT), k{sub cat} remains nearly constant at 13 min{sup {minus}1}. However, k{sub cat}/k{sub m} with nuclease wt varies considerably. The data suggests that the inflection k{sub cat}/K{sub m} with pK{sub a} at 9.67 arises from ionization of tyrosine-85, which hydrogen bonds to the divalent 3'-phosphomonester of substrates with this substituent. The enthalpy of ionization of both deprotonation steps in the k{sub cat}/K{sub m} versus pH profile is 5 kcal/mol. {sup 13}C NMR has been used to determine the pK{sub a} values of the arginine and tyrosine residues. The results do not rule out arginine as a candidate for the acidic catalyst that protonates the 5'-ribose alkoxide prior to product release. The phenolic hydroxyl carbon of tyrosine-85 has been assigned by comparing the {sup 13}C NMR spectrum of nuclease wt and nuclease Y85F. This correlation between pK{sub a} values along with the absence of other candidates indicates that the ionization of tyrosine-85 is the pK{sub a} seen in the k{sub cat}/K{sub m} vs pH profile for substrates with a divalent 3'-phosphomonester. This conclusion is consistent with the proposed role of tyrosine-85 as a hydrogen-bond donor to the 3'-phosphomonoester of substrates poised for exonucleolytic hydrolysis.

  19. Plant Habitat (PH)

    NASA Technical Reports Server (NTRS)

    Onate, Bryan

    2016-01-01

    The International Space Station (ISS) will soon have a platform for conducting fundamental research of Large Plants. Plant Habitat (PH) is designed to be a fully controllable environment for high-quality plant physiological research. PH will control light quality, level, and timing, temperature, CO2, relative humidity, and irrigation, while scrubbing ethylene. Additional capabilities include leaf temperature and root zone moisture and oxygen sensing. The light cap will have red (630 nm), blue (450 nm), green (525 nm), far red (730 nm) and broad spectrum white LEDs. There will be several internal cameras (visible and IR) to monitor and record plant growth and operations.

  20. Field solar degradation of pesticides and emerging water contaminants mediated by polymer films containing titanium and iron oxide with synergistic heterogeneous photocatalytic activity at neutral pH.

    PubMed

    Mazille, F; Schoettl, T; Klamerth, N; Malato, S; Pulgarin, C

    2010-05-01

    Photocatalytic degradation of phenol, nalidixic acid, mixture of pesticides, and another of emerging contaminants in water was mediated by TiO(2) and iron oxide immobilized on functionalized polyvinyl fluoride films (PVF(f)-TiO(2)-Fe oxide) in a compound parabolic collector (CPC) solar photoreactor. During degradation, little iron leaching (<0.2mgL(-1)) was observed. Phenol was efficiently degraded and mineralized at operational pH<5 and nalidixic acid degradation was complete even at pH 7, but mineralization stopped at 35%. Pesticide mixture was slowly degraded (50%) after 150min of irradiation. Degradation of the emergent contaminant mixture was successful for eight compounds and less efficient for six other compounds. The significant reactivity differences between tested compounds were assigned to the differences in structure namely that the presence of complexing or chelating groups enhanced the rates. PVF(f)-TiO(2)-Fe oxide photoactivity gradually increased during 20 days of experiments. X-ray photoelectron spectroscopy (XPS) measurements revealed significant changes on the catalyst surface. These analyses confirm that during photocatalysis mediated by PVF(f)-TiO(2)-Fe oxide, some iron leaching led to enlargement of the TiO(2) surface exposed to light, increasing its synergy with iron oxides and leading to enhanced pollutant degradation.

  1. pH Basics

    ERIC Educational Resources Information Center

    Lunelli, Bruno; Scagnolari, Francesco

    2009-01-01

    The exposition of the pervasive concept of pH, of its foundations and implementation as a meaningful quantitative measurement, in nonspecialist university texts is often not easy to follow because too many of its theoretical and operative underpinnings are neglected. To help the inquiring student we provide a concise introduction to the depth just…

  2. pH optrode

    DOEpatents

    Northrup, M. Allen; Langry, Kevin C.

    1993-01-01

    A process is provided for forming a long-lasting, stable, pH-sensitive dye-acrylamide copolymer useful as a pH-sensitive material for use in an optrode or other device sensitive to pH. An optrode may be made by mechanically attaching the copolymer to a sensing device such as an optical fiber.

  3. Ph.D. shortage

    NASA Astrophysics Data System (ADS)

    The late 1990s will see a shortage of Ph.D. graduates, according to the Association of American Universities, Washington, D.C. AAU's new comprehensive study, “The Ph.D. Shortage: The Federal Role,” reports that competition for new Ph.D.s is already intense and can only intensify because demand is greater than supply in both academic and nonacademic markets.Doctoral education plays an increasingly important role in U.S. research and development programs. Students have a pivotal part in doing research and enriching it with new ideas. The AAU report says that graduate students are “major determinants of the creativity and productivity of U.S. academic research, the source of more than 50% of the nation's basic research.’ The market for doctoral education extends beyond the university. In 1985, about 43% of all Ph.D.s employed in this country were working outside higher education; the demand for doctorate recipients in nonacademic sectors continues to grow.

  4. Evaluation of sample preparation methods for water activity determination in jerky and kippered beef: a research note.

    PubMed

    Harper, N M; Getty, K J K; Boyle, E A E

    2010-10-01

    Commercially available packaged whole muscle beef jerky, chopped and formed beef jerky, and kippered beef steak were obtained from retail stores to determine the effect of two sample preparation methods on water activity (a(w)). Intact samples were prepared by cutting product into a hexagonal shape with a 3.2 cm diameter. Diced samples were prepared by cutting the product into 0.4 x 0.4 cm squares. Whole muscle jerky a(w) was higher (0.016 units; P<0.001) using the intact method compared to the diced method. There was no difference (P>0.05) in a(w) levels between the two preparation methods for chopped and formed jerky or kippered beef steak. An intact sample preparation method is recommended for a(w) determination of whole muscle jerky to obtain a more conservative value, especially if a(w) is near the margin of safety.

  5. Water activity and temperature effects on growth of Eurotium amstelodami, E. chevalieri and E. herbariorum on a sponge cake analogue.

    PubMed

    Abellana, M; Magrí, X; Sanchis, V; Ramos, A J

    1999-11-01

    Eurotium is a widespread storage fungal genus that has been frequently isolated from bakery products. The objective of this study was (i) to obtain a method for studying the growth of xerophilic fungi on bakery products, and (ii) to determine the effects of water activity (a(w)), temperature, isolate and their interactions on mycelial growth of Eurotium spp. on an analogue medium of sponge cake. Statistical analysis showed that there were intra-isolate differences (P<0.001) due to a(w), temperature, isolate, and two- and three-way interactions. Optimum growth of all isolates over a(w) x temperature range tested showed optima at 0.90 a(w) and 30 degrees C, with an interval of growth rate of 3.8-5.1 mm x d(-1). At 0.75 a(w), growth was less than 0.15 mm x d(-1), if there was any.

  6. The effect of NaCl substitution with KCl on proteinase activities of cell-free extract and cell-free supernatant at different pH levels and salt concentrations: Lactobacillus acidophilus and Lactobacillus casei.

    PubMed

    Ayyash, M M; Sherkat, F; Shah, N P

    2013-01-01

    The aim of this study was to investigate the effect of substitution of NaCl with KCl at different pH levels and salt concentrations on proteinase activity of cell-free extract and cell-free supernatant of the probiotics Lactobacillus acidophilus and Lactobacillus casei. de Man, Rogosa, and Sharpe broth aliquots were mixed with 2 pure salts (NaCl and KCl) and 2 salt concentrations at 2 concentration levels (5 and 10%), inoculated with Lactobacillus acidophilus or Lactobacillus casei, and incubated aerobically at 37°C for 22 h. The cultures were then centrifuged at 4,000×g for 30 min, and the collected cell pellets were used to prepare cell-wall proteinases and the supernatants used as a source of supernatant (extracellular) proteinases. The proteolytic activity and protein content of both portions were determined. After incubation of both portions with 3 milk caseins (α-, β-, κ-casein), the supernatants were individually subjected to analysis of angiotensin-converting enzyme (ACE)-inhibitory activity and proteolytic activity using the o-phthalaldehyde method. Significant differences were observed in ACE-inhibitory and proteolytic activities between salt substitution treatments of cell-free extract and cell-free supernatant from both probiotic strains at the same salt concentration and pH level.

  7. Running climate model on a commercial cloud computing environment: A case study using Community Earth System Model (CESM) on Amazon AWS

    NASA Astrophysics Data System (ADS)

    Chen, Xiuhong; Huang, Xianglei; Jiao, Chaoyi; Flanner, Mark G.; Raeker, Todd; Palen, Brock

    2017-01-01

    The suites of numerical models used for simulating climate of our planet are usually run on dedicated high-performance computing (HPC) resources. This study investigates an alternative to the usual approach, i.e. carrying out climate model simulations on commercially available cloud computing environment. We test the performance and reliability of running the CESM (Community Earth System Model), a flagship climate model in the United States developed by the National Center for Atmospheric Research (NCAR), on Amazon Web Service (AWS) EC2, the cloud computing environment by Amazon.com, Inc. StarCluster is used to create virtual computing cluster on the AWS EC2 for the CESM simulations. The wall-clock time for one year of CESM simulation on the AWS EC2 virtual cluster is comparable to the time spent for the same simulation on a local dedicated high-performance computing cluster with InfiniBand connections. The CESM simulation can be efficiently scaled with the number of CPU cores on the AWS EC2 virtual cluster environment up to 64 cores. For the standard configuration of the CESM at a spatial resolution of 1.9° latitude by 2.5° longitude, increasing the number of cores from 16 to 64 reduces the wall-clock running time by more than 50% and the scaling is nearly linear. Beyond 64 cores, the communication latency starts to outweigh the benefit of distributed computing and the parallel speedup becomes nearly unchanged.

  8. Revitalisation As a Force Promoting Spatial Changes in Wrocław/ Idea Rewitalizacji Motorem Przemian Przestrzennych Wrocławia

    NASA Astrophysics Data System (ADS)

    Kożan, Aldona

    2015-06-01

    The idea of revitalization, which grew out of the need to repair degraded urban spaces, has taken on a new meaning in the context of contemporary development challenges. The work includes references to the basic programme documents and strategies. It shows regeneration activities carried out in Wrocław in connection with the organization of international events, cultural and sport events, and with the improvement of spatial order. These actions contributed to the revitalization of the image of the city. The paper contains the results of these changes, the complementarity of decisions and their impact on social behaviour. Idea rewitalizacji, wyrosła z konieczności naprawy zdegradowanych przestrzeni miejskich i zniesienia konfliktów społecznych, nabrała nowego znaczenia w kontekście współczesnych wyzwań rozwojowych. Władze Wrocławia opracowały strategię i szereg dokumentów programowych, których efektem są zrealizowane przedsięwzięcia rewitalizacyjne: obiekty stanowiące infrastrukturę techniczną dla realizacji imprez sportowo - kulturalnych (Stadion Miejski Euro 2012, obiekty Hali Stulecia, rozwoju nauki: kampusy uczelni wyższych i obiekty kultury). Rewitalizacji przestrzennej zostały poddane przestrzenie publiczne miasta: ulice i place, modernizacji i rozbudowie podlegały układy i obiekty transportowe: nowy terminal lotniczy, przebudowa Dworca Głównego we Wrocławiu. Działania rewitalizacyjne wpłynęły na nowy wizerunek miasta, podniosły jakość życia i wyzwoliły nowe zachowania społeczne. Praca zawiera wyniki tych przemian, komplementarności podjętych decyzji i ich wpływu na zachowania społeczne.

  9. The influence of redox chemistry and pH on chemically active forms of arsenic in sewage sludge-amended soil

    SciTech Connect

    Carbonell-Barrachina, A.; Jugsujinda, A.; DeLaune, R.D.; Patrick, W.H. Jr.; Burlo, F.; Sirisukhodom, S.; Anurakpongsatorn, P.

    1999-07-01

    Chemical fractionation procedures were used to quantify the effect of the sediment redox and pH conditions on the adsorption and solubility of arsenic (As) in municipal sewage sludge and sewage sludge-amended soil. Sludge and sludge-amended soil were incubated in microcosms in which Eh-pH conditions were controlled. Samples were sequentially extracted to determine As in various chemical forms (water soluble, exchangeable, bound to carbonates, bound to iron (Fe) and manganese (Mn) oxides, bound to insoluble organics and sulfides) and the chemically inactive fraction (mineral residues). In both sewage sludge and sludge-amended soil, As chemistry was governed by large molecular humic matter and sulfides and Fe and Mn-oxides. Solubility of As remained low and constant under both aerobic and anaerobic conditions in sludge-amended soil. After dissolution of Fe and Mn-oxides, As{sup 5+} was released into sludge solution, reduced to As{sup 3+} and likely precipitated as sulfide. Therefore, an organic amendment rich in sulfur compounds, such as sewage sludge, would drastically reduce the potential risks derived from As pollution under highly anoxic conditions by precipitation of this toxic metalloid as insoluble and immobile sulfides.

  10. The Activity of TcCYS4 Modified by Variations in pH and Temperature Can Affect Symptoms of Witches’ Broom Disease of Cocoa, Caused by the Fungus Moniliophthora perniciosa

    PubMed Central

    Freitas, Ana Camila Oliveira; Souza, Cristiane Ferreira; Monzani, Paulo Sérgio; Garcia, Wanius; de Almeida, Alex Alan Furtado; Costa, Marcio Gilberto Cardoso; Pirovani, Carlos Priminho

    2015-01-01

    The phytocystatins regulate various physiological processes in plants, including responses to biotic and abiotic stresses, mainly because they act as inhibitors of cysteine proteases. In this study, we have analyzed four cystatins from Theobroma cacao L. previously identified in ESTs libraries of the interaction with the fungus Moniliophthora perniciosa and named TcCYS1, TcCYS2, TcCYS3 and TcCYS4. The recombinant cystatins were purified and subjected to the heat treatment, at different temperatures, and their thermostabilities were monitored using their ability to inhibit papain protease. TcCYS1 was sensitive to temperatures above 50°C, while TcCYS2, TcCYS3, and TcCYS4 were thermostable. TcCYS4 presented a decrease of inhibitory activity when it was treated at temperatures between 60 and 70°C, with the greater decrease occurring at 65°C. Analyses by native gel electrophoresis and size-exclusion chromatography showed that TcCYS4 forms oligomers at temperatures between 60 and 70°C, condition where reduction of inhibitory activity was observed. TcCYS4 oligomers remain stable for up to 20 days after heat treatment and are undone after treatment at 80°C. TcCYS4 presented approximately 90% of inhibitory activity at pH values between 5 and 9. This protein treated at temperatures above 45°C and pH 5 presented reduced inhibitory activity against papain, suggesting that the pH 5 enhances the formation of TcCYS4 oligomers. A variation in the titratable acidity was observed in tissues of T. cacao during the symptoms of witches’ broom disease. Our findings suggest that the oligomerization of TcCYS4, favored by variations in pH, is an endergonic process. We speculate that this process can be involved in the development of the symptoms of witches’ broom disease in cocoa. PMID:25830226

  11. A highly efficient Mukaiyama-Mannich reaction of N-Boc isatin ketimines and other active cyclic ketimines using difluoroenol silyl ethers catalyzed by Ph3PAuOTf.

    PubMed

    Yu, Jin-Sheng; Zhou, Jian

    2015-12-07

    Ph3PAuOTf is identified as a powerful catalyst for the addition of difluoroenol silyl ethers to N-Boc isatin ketimines and other two kinds of active cyclic ketimines. This represents the first Au(i)-catalyzed Mukaiyama-Mannich reaction, and the corresponding non-fluorinated enol silyl ether proves to be even much more reactive under the same conditions. This method paves the way to the total synthesis of difluoromethylated analogues of AG-041R, a gastrin/CCK-B receptor antagonist.

  12. 17-4 PH and 15-5 PH

    NASA Technical Reports Server (NTRS)

    Johnson, Howard T.

    1995-01-01

    17-4 PH and 15-5 PH are extremely useful and versatile precipitation-hardening stainless steels. Armco 17-4 PH is well suited for the magnetic particle inspection requirements of Aerospace Material Specification. Armco 15-5 PH and 17-4 PH are produced in billet, plate, bar, and wire. Also, 15-5 PH is able to meet the stringent mechanical properties required in the aerospace and nuclear industries. Both products are easy to heat treat and machine, making them very useful in many applications.

  13. LiDAR and 2D Electrical Resistivity Tomography as a Supplement of Geomorphological Investigations in Urban Areas: a Case Study from the City of Wrocław (SW Poland)

    NASA Astrophysics Data System (ADS)

    Kasprzak, Marek; Traczyk, Andrzej

    2014-06-01

    In urbanized areas, particularly in lowland terrains and floors of large river valleys, the natural land configuration is often hard to recognize due to a long history of human activity. Accordingly, archaeological works in cities, which supply knowledge on settlement conditions, are usually accompanied by geological and geomophological research. Lately, data from light detection and ranging (LiDAR) have become a valuable source of information on urban land configuration. Geophysical methods are also becoming increasingly popular in background studies. The paper presents a method of using and linking these sources of spatial information about landforms in such areas. The main aim is to identify to what extent these complementary sources of data and the proposed method can be used in such a specific environment to reconstruct natural, buried terrain morphology. The city of Wrocław in Central Europe serves as an example. To this end geomorphometric studies were conducted with the use of digital elevation models (DEMs) based on LiDAR scanning and derivated land-surface parameters—SAGA Wetness Index, Channel Network Base Level and Altitude above Channel Network. The study also involved determining morphological edges and measurements of the meanders of the Odra, as well as expanding information on the spatial distribution of alluvia and the structure of slope breaks. To this end, geophysical measurements were conducted using the Two-Dimensional Electrical Resistivity Tomography method. Additionally, five typical sequences of man-made ground present within the perimeter of the city were distinguished. As a result, a map of the main landforms of Wrocław is presented. Finally, we argue that although high resolution DEM and derivate land-surface parameters are very useful in terrain analysis, places with thick man-made ground or strongly levelled areas must be recognized by geoarchaeological excavations or geological bore holes. The geophysical survey is useful to

  14. PET Imaging of Extracellular pH in Tumors with 64Cu- and 18F-Labeled pHLIP Peptides: A Structure–Activity Optimization Study

    PubMed Central

    2016-01-01

    pH (low) insertion peptides (pHLIP peptides) target acidic extracellular environments in vivo due to pH-dependent cellular membrane insertion. Two variants (Var3 and Var7) and wild-type (WT) pHLIP peptides have shown promise for in vivo imaging of breast cancer. Two positron emitting radionuclides (64Cu and 18F) were used to label the NOTA- and NO2A-derivatized Var3, Var7, and WT peptides for in vivo biodistribution studies in 4T1 orthotopic tumor-bearing BALB/c mice. All of the constructs were radiolabeled with 64Cu or [18F]-AlF in good yield. The in vivo biodistribution of the 12 constructs in 4T1 orthotopic allografted female BALB/c mice indicated that NO2A-cysVar3, radiolabeled with either 18F (4T1 uptake; 8.9 ± 1.7%ID/g at 4 h p.i.) or 64Cu (4T1 uptake; 8.2 ± 0.9%ID/g at 4 h p.i. and 19.2 ± 1.8% ID/g at 24 h p.i.), shows the most promise for clinical translation. Additional studies to investigate other tumor models (melanoma, prostate, and brain tumor models) indicated the universality of tumor targeting of these tracers. From this study, future clinical translation will focus on 18F- or 64Cu-labeled NO2A-cysVar3. PMID:27396694

  15. Time-dependent activity of Na+/H+ exchanger isoform 1 and homeostasis of intracellular pH in astrocytes exposed to CoCl2 treatment.

    PubMed

    Wang, Peng; Li, Ling; Zhang, Zhenxiang; Kan, Quancheng; Gao, Feng; Chen, Suyan

    2016-05-01

    Hypoxia causes injury to the central nervous system during stroke and has significant effects on pH homeostasis. Na+/H+ exchanger isoform 1 (NHE1) is important in the mechanisms of hypoxia and intracellular pH (pHi) homeostasis. As a well-established hypoxia-mimetic agent, CoCl2 stabilizes and increases the expression of hypoxia inducible factor‑1α (HIF-1α), which regulates several genes involved in pH balance, including NHE1. However, it is not fully understood whether NHE1 is activated in astrocytes under CoCl2 treatment. In the current study, pHi and NHE activity were analyzed using the pHi‑sensitive dye BCECF‑AM. Using cariporide (an NHE1‑specific inhibitor) and EIPA (an NHE nonspecific inhibitor), the current study demonstrated that it was NHE1, not the other NHE isoforms, that was important in regulating pHi homeostasis in astrocytes during CoCl2 treatment. Additionally, the present study observed that, during the early period of CoCl2 treatment (the first 2 h), NHE1 activity and pHi dropped immediately, and NHE1 mRNA expression was reduced compared with control levels, whereas expression levels of the NHE1 protein had not yet changed. In the later period of CoCl2 treatment, NHE1 activity and pHi significantly increased compared with the control levels, as did the mRNA and protein expression levels of NHE1. Furthermore, the cell viability and injury of astrocytes was not changed during the initial 8 h of CoCl2 treatment; their deterioration was associated with the higher levels of pHi and NHE1 activity. The current study concluded that NHE1 activity and pHi homeostasis are regulated by CoCl2 treatment in a time-dependent manner in astrocytes, and may be responsible for the changes in cell viability and injury observed under hypoxia-mimetic conditions induced by CoCl2 treatment.

  16. Comparisons of the Pentax-AWS, Glidescope, and Macintosh Laryngoscopes for Intubation Performance during Mechanical Chest Compressions in Left Lateral Tilt: A Randomized Simulation Study of Maternal Cardiopulmonary Resuscitation

    PubMed Central

    Lee, Sanghyun; Kim, Wonhee; Kang, Hyunggoo; Oh, Jaehoon; Lim, Tae Ho; Lee, Yoonjae; Kim, Changsun; Cho, Jun Hwi

    2015-01-01

    Purpose. Rapid advanced airway management is important in maternal cardiopulmonary resuscitation (CPR). This study aimed to compare intubation performances among Pentax-AWS (AWS), Glidescope (GVL), and Macintosh laryngoscope (MCL) during mechanical chest compression in 15° and 30° left lateral tilt. Methods. In 19 emergency physicians, a prospective randomized crossover study was conducted to examine the three laryngoscopes. Primary outcomes were the intubation time and the success rate for intubation. Results. The median intubation time using AWS was shorter than that of GVL and MCL in both tilt degrees. The time to visualize the glottic view in GVL and AWS was significantly lower than that of MCL (all P < 0.05), whereas there was no significant difference between the two video laryngoscopes (in 15° tilt, P = 1; in 30° tilt, P = 0.71). The progression of tracheal tube using AWS was faster than that of MCL and GVL in both degrees (all P < 0.001). Intubations using AWS and GVL showed higher success rate than that of Macintosh laryngoscopes. Conclusions. The AWS could be an appropriate laryngoscope for airway management of pregnant women in tilt CPR considering intubation time and success rate. PMID:26161426

  17. Crystal structures of cyanide- and triiodide-bound forms of Arthromyces ramosus peroxidase at different pH values. Perturbations of active site residues and their implication in enzyme catalysis.

    PubMed

    Fukuyama, K; Kunishima, N; Amada, F; Kubota, T; Matsubara, H

    1995-09-15

    The structures of the cyanide and triiodide complexes of Arthromyces ramosus peroxidase (ARP) at different pH values were investigated by x-ray crystallography in order to examine the behavior of the invariant residues of arginine (Arg-52) and distal histidine (His-56) during the enzyme reaction as well as to provide the structural basis of the active site of peroxidase. The models of the cyanide complexes at pH 7.5, 5.0, and 4.0, respectively, were refined to the R-factors of 17.8, 17.8, and 18.5% using 7.0-1.6-A resolution data, and those of the triiodide complexes at pH 6.5 and 5.0 refined to 16.9 and 16.8% using 7.0-1.9-A resolution data. The structures of the cyanide complexes at pH 7.5, 5.0, and 4.0 are identical within experimental error. Cyanide ion bound to the heme in the bent conformation rather than in the tilt conformation. Upon cyanide ion binding, the N epsilon atom of His-56 moved toward the ion by rotation of the imidazole ring around the C beta-C gamma bond, but there was little conformational change in the remaining residues. The distance between the N epsilon atom of His-56 and the nitrogen atom of the cyanide suggests the presence of a hydrogen bond between them in the pH range investigated. In the triiodide complexes, one of the two triiodides bound to ARP was located at the distal side of the heme. When triiodide bound to ARP, unlike the rearrangement of the distal arginine of cytochrome c peroxidase that occurs on formation of the fluoride complex or compound I, the side chain of Arg-52 moved little. The conformation of the side chain of His-56, however, changed markedly. Conformational flexibility of His-56 appears to be a requisite for proton translocation from one oxygen atom to the other of HOO- by acid-base catalysis to produce compound I. The iron atom in each cyanide complex (low-spin ferric) is located in the heme plane, whereas in each triiodide complex (high-spin ferric) the iron atom is displaced from the plane about 0.2 A toward

  18. [Some remarks on the theory of sets by Richard Dedekind and Stanisław Leśniewski].

    PubMed

    Obojska, Lidia

    2014-01-01

    Mereology, is a part-whole theory, also called the theory of collective sets. It was founded in 1916 by Stanisław Leśniewski and this is an alternative theory versus the classical set theory by Georg Cantor. These two theories are usually teamed up together as Leśniewski himself was referring to the concept of the set by Cantor and Cantor is considered the "main" ideologist of the set theory. However, when analyzing the original texts of various authors, it seems that the very concept of a collective set is closer to the idea of Richard Dedekind rather than that of Georg Cantor. It is known that Cantor borrowed some concepts on the notion of set from Dedekind, whose ideas were also known to Leśniewski, however, there is no study on this topic. This work is therefore an attempt to compare some set-theoretical concepts of both of these authors, i.e. S. Leśiewski and R. Dedekind and the presentation of their convergence.

  19. Activation of Aplysia ARF6 induces neurite outgrowth and is sequestered by the overexpression of the PH domain of Aplysia Sec7 proteins.

    PubMed

    Jang, Deok-Jin; Jun, Yong-Woo; Shim, Jaehoon; Sim, Su-Eon; Lee, Jin-A; Lim, Chae-Seok; Kaang, Bong-Kiun

    2017-02-01

    ADP-ribosylation factors (ARFs) are small guanosine triphosphatases of the Ras superfamily involved in membrane trafficking and regulation of the actin cytoskeleton. Aplysia Sec7 protein (ApSec7), a guanine nucleotide exchange factor for ARF1 and ARF6, induces neurite outgrowth and plays a key role in 5-hydroxyltryptamine-induced neurite growth and synaptic facilitation in Aplysia sensory-motor synapses. However, the specific role of ARF6 signaling on neurite outgrowth in Aplysia neurons has not been examined. In the present study, we cloned Aplysia ARF6 (ApARF6) and revealed that an overexpression of enhanced green fluorescent protein (EGFP)-fused constitutively active ApARF6 (ApARF6-Q67L-EGFP) could induce neurite outgrowth in Aplysia sensory neurons. Further, we observed that ApARF6-induced neurite outgrowth was inhibited by the co-expression of a Sec7 activity-deficient mutant of ApSec7 (ApSec7-E159K). The pleckstrin homology domain of ApSec7 may bind to active ApARF6 at the plasma membrane and prevent active ApARF6-induced functions, including intracellular vacuole formation in HEK293T cells. The results of the present study suggest that activation of ARF6 signaling could induce neurite outgrowth in Aplysia neurons and may be involved in downstream signaling of ApSec7-induced neurite outgrowth in Aplysia neurons.

  20. Proven in vitro evolution of protease cathepsin E-inhibitors and -activators at pH 4.5 using a paired peptide method.

    PubMed

    Kitamura, Koichiro; Komatsu, Masayuki; Biyani, Madhu; Futakami, Masae; Kawakubo, Tomoyo; Yamamoto, Kenji; Nishigaki, Koichi

    2012-12-01

    Improving a particular function of molecules is often more difficult than identifying such molecules ab initio. Here, a method to acquire higher affinity and/or more functional peptides was developed as a progressive library selection method. The primary library selection products were utilized to build a secondary library composed of blocks of 4 amino acids, of which selection led to peptides with increased activity. These peptides were further converted to randomly generate paired peptides. Cathepsin E-inhibitors thus obtained exhibited the highest activities and affinities (pM order). This was also the case with cathepsin E-activating peptides, proving the methodological effectiveness. The primary, secondary, and tertiary library selections can be regarded as module-finding, module-shuffling, and module-pairing, respectively, which resembles the progression of the natural evolution of proteins. The mode of peptide binding to their target proteins is discussed in analogy to antibodies and epitopes of an antigen.

  1. Sono-incorporation of CuO nanoparticles on the surface and into the mesoporous hexatitanate layers: Enhanced Fenton-like activity in degradation of orange-G at its neutral pH

    NASA Astrophysics Data System (ADS)

    Sehati, S.; Entezari, M. H.

    2017-03-01

    In this work, for the first time, CuO/Ti6O13 mesoporous nanocomposite was synthesized by direct intercalation of CuO nanoparticles into hexatitanate layers in the presence of ultrasound. In fact mesoporous potassium hexatitanate with high pore size (44.94 nm) was used as a support. CuO nanoparticles were grown into the titanate pores which caused formation of ultra small CuO with uniform size and high surface area. In fact, titanate is considered as a substrate for better dispersion and nucleation of the CuO nanoparticles which prevented the agglomeration and overgrowth of guest molecules. The prepared sample was characterized by XRD, FE-SEM, TEM, UV-vis spectra, N2 adsorption-desorption, Raman spectra and FT-IR techniques. The product was used as a heterogeneous Fenton-like catalyst for the degradation of Orang G (OG). The effect of important parameters, including pH, H2O2 addition rate and catalyst loading on the decolorization of OG were investigated. Based on the results, CuO/Ti6O13 catalyst exhibited high catalytic activity for OG degradation in aqueous solution at neutral pH of the dye. Moreover, breaking of H2O2 during the catalytic reaction was monitored by spectroscopic method. The results confirmed the decomposition of H2O2 to produce rad OH which is the main active species for the degradation of OG.

  2. Water activity and temperature effects on growth of Alternaria arborescens on tomato medium.

    PubMed

    Vaquera, Sandra; Patriarca, Andrea; Fernández Pinto, Virginia

    2014-08-18

    Alternaria arborescens is the causal agent of tomato stem canker, a disease frequently responsible of substantial economic losses. A. arborescens can produce several mycotoxins, such as alternariol, alternariol monomethyl ether and tenuazonic acid and phytotoxins such as the AAL toxins. The objective of this study was to determine the effect of water activity (aw, 0.950, 0.975, 0.995) and temperature (6, 15, 20, 25 and 30°C) on the germination and radial growth rate of A. arborescens on a synthetic tomato medium. Germination followed by growth was observed at all temperatures and aw levels analyzed. The shortest germination time (0.5 days) was observed at 0.995 aw, both at 25°C and at 30°C. The germination time increased with a reduction of aw and temperature. The highest growth rate was registered at 0.995 aw and 30°C (7.21 mm/day) while the lowest occurred at 0.950 aw and 6°C (0.52 mm/day), conditions at which the longest lag phase was observed (8 days). Growth rates increased with aw and temperature. Knowledge of the ecophysiology of the fungus in this substrate is necessary to formulate future strategies to prevent its development and evaluate the consumer health risk posed by potential exposure to the toxins.

  3. Chemical composition, antimicrobial and antioxidant activities of the essential oils of Sideritis erythrantha Boiss. and Heldr. (var. erythrantha and var. cedretorum P.H. Davis) endemic in Turkey.

    PubMed

    Köse, Elif Odabaş; Deniz, Ismail Gökhan; Sarıkürkçü, Cengiz; Aktaş, Ozgür; Yavuz, Mustafa

    2010-10-01

    In the present study, chemical compositions, antimicrobial and antioxidant activities of the essential oils of Sideritis erythrantha var. erythrantha (SE) and Sideritis erythrantha var. cedretorum (SC), which are endemic taxa in Turkey, were investigated. The essential oils obtained by hydrodistillation were analyzed by gas chromatography-mass spectrometry (GC-MS). α-Pinene was the major component of the essential oils of SC and SE. SC essential oil was as effective as antibiotic against methicillin-resistant Staphylococcus aureus (MRSA), vancomycin resistant Enterococcus faecalis (VRE), ampicillin resistant Haemophilusinfluenzae and vancomycin sensitive E. faecalis. Similarly, SE essential oil was also as effective as antibiotic against VRE and ampicillin resistant H. influenzae. Antioxidant activities of the essential oils of SC and SE were determined by 1,1-diphenyl-2-picrylhydrazyl (DPPH), β-carotene/linoleic acid and reducing power. Both essential oils exhibited weak antioxidant activity. This is the first report on antimicrobial and antioxidant activities of the essential oils of SC and SE.

  4. Integrated Activity and Genetic Profiling of Secreted Peptidases in Cryptococcus neoformans Reveals an Aspartyl Peptidase Required for Low pH Survival and Virulence

    PubMed Central

    Clarke, Starlynn C.; Dumesic, Phillip A.; Homer, Christina M.; O’Donoghue, Anthony J.; La Greca, Florencia; Pallova, Lenka; Majer, Pavel; Madhani, Hiten D.; Craik, Charles S.

    2016-01-01

    The opportunistic fungal pathogen Cryptococcus neoformans is a major cause of mortality in immunocompromised individuals, resulting in more than 600,000 deaths per year. Many human fungal pathogens secrete peptidases that influence virulence, but in most cases the substrate specificity and regulation of these enzymes remains poorly understood. The paucity of such information is a roadblock to our understanding of the biological functions of peptidases and whether or not these enzymes are viable therapeutic targets. We report here an unbiased analysis of secreted peptidase activity and specificity in C. neoformans using a mass spectrometry-based substrate profiling strategy and subsequent functional investigations. Our initial studies revealed that global peptidase activity and specificity are dramatically altered by environmental conditions. To uncover the substrate preferences of individual enzymes and interrogate their biological functions, we constructed and profiled a ten-member gene deletion collection of candidate secreted peptidases. Through this deletion approach, we characterized the substrate specificity of three peptidases within the context of the C. neoformans secretome, including an enzyme known to be important for fungal entry into the brain. We selected a previously uncharacterized peptidase, which we term Major aspartyl peptidase 1 (May1), for detailed study due to its substantial contribution to extracellular proteolytic activity. Based on the preference of May1 for proteolysis between hydrophobic amino acids, we screened a focused library of aspartyl peptidase inhibitors and identified four high-affinity antagonists. Finally, we tested may1Δ strains in a mouse model of C. neoformans infection and found that strains lacking this enzyme are significantly attenuated for virulence. Our study reveals the secreted peptidase activity and specificity of an important human fungal pathogen, identifies responsible enzymes through genetic tests of their

  5. H⁺-activated Na⁺ influx in the ventricular myocyte couples Ca²⁺-signalling to intracellular pH.

    PubMed

    Garciarena, Carolina D; Youm, Jae Boum; Swietach, Pawel; Vaughan-Jones, Richard D

    2013-08-01

    Acid extrusion on Na(+)-coupled pH-regulatory proteins (pH-transporters), Na(+)/H(+) exchange (NHE1) and Na(+)-HCO3(-) co-transport (NBC), drives Na(+) influx into the ventricular myocyte. This H(+)-activated Na(+)-influx is acutely up-regulated at pHi<7.2, greatly exceeding Na(+)-efflux on the Na(+)/K(+) ATPase. It is spatially heterogeneous, due to the co-localisation of NHE1 protein (the dominant pH-transporter) with gap-junctions at intercalated discs. Overall Na(+)-influx via NBC is considerably lower, but much is co-localised with L-type Ca(2+)-channels in transverse-tubules. Through a functional coupling with Na(+)/Ca(2+) exchange (NCX), H(+)-activated Na(+)-influx increases sarcoplasmic-reticular Ca(2+)-loading and release during in