Optimization of Culture Medium Enhances Viable Biomass Production and Biocontrol Efficacy of the Antagonistic Yeast, Candida diversa

PubMed Central

Liu, Jia; Li, Guangkun; Sui, Yuan

2017-01-01

Viable biomass production is a key determinant of suitability of antagonistic yeasts as potential biocontrol agents. This study investigated the effects of three metal ions (magnesium, ferrous, and zinc) on biomass production and viability of the antagonistic yeast, Candida diversa. Using response surface methodology to optimize medium components, a maximum biomass was obtained, when the collective Mg2+, Fe2+, and Zn2+ concentrations were adjusted in a minimal mineral (MM) medium. Compared with the unmodified MM, and three ion-deficient MM media, yeast cells cultured in the three ion-modified MM medium exhibited a lower level of cellular oxidative damage, and a higher level of antioxidant enzyme activity. A biocontrol assay indicated that C. diversa grown in the ion-modified MM exhibited the greatest level of control of gray mold on apple fruit. These results provide new information on culture medium optimization to grow yeast antagonists in order to improve biomass production and biocontrol efficacy. PMID:29089939

The effects of cyclosporin on the collagenolytic activity of gingival fibroblasts.

PubMed

Hyland, Paula L; Traynor, Patrick S; Myrillas, Theofilos T; Marley, John J; Linden, Gerard J; Winter, Paul; Leadbetter, Nicola; Cawston, Timothy E; Irwin, Chris R

2003-04-01

The immunosuppressive agent cyclosporin is associated with a number of major side-effects including the development of gingival overgrowth. Although the pathogenesis of cyclosporin-induced gingival overgrowth remains unclear, it has been suggested that the finely regulated balance between extracellular matrix synthesis and degradation may be disturbed, resulting in an accumulation of excess connective tissue components within the gingival tissue. The aim of this study was to investigate the effect of cyclosporin on matrix metalloproteinases (MMP)-1 and tissue inhibitors of MMP (TIMP)-1 expression at the mRNA, protein, and enzyme activity levels. Gingival fibroblasts were grown to confluence and then cultured in serum-free medium supplemented with cyclosporin over the concentration range of 0 to 2000 ng/ml. MMP-1 and TIMP-1 mRNA levels in cultures were determined by reverse transcription polymerase chain reaction (RT-PCR), protein levels in whole conditioned medium were assessed by enzyme-linked immunosorbent assay (ELISA), and collagenolytic activity determined using a 3H-acetylated type I collagen degradation assay. Tissue mRNA levels in normal and overgrown gingiva were also determined by RT-PCR. Results indicated that cyclosporin inhibited MMP-1 expression at both the mRNA and protein level in a dose- and time-dependent fashion. The effects on TIMP-1 expression were less clear, cyclosporin inhibiting mRNA expression, but having no effect on TIMP-1 protein levels at any concentration studied. Addition of the drug resulted in reduced levels of collagenolytic activity in the culture medium. MMP-1 mRNA expression was significantly reduced in overgrown compared to normal tissue. These results add support to the hypothesis that the accumulation of collagen seen in gingival overgrowth can be explained by a cyclosporin-induced inhibition of collagenolytic activity within the gingival tissues.

Activation of the prelimbic medial prefrontal cortex induces anxiety-like behaviors via N-Methyl-D-aspartate receptor-mediated glutamatergic neurotransmission in mice.

PubMed

Saitoh, Akiyoshi; Ohashi, Masanori; Suzuki, Satoshi; Tsukagoshi, Mai; Sugiyama, Azusa; Yamada, Misa; Oka, Jun-Ichiro; Inagaki, Masatoshi; Yamada, Mitsuhiko

2014-08-01

We investigated the possible roles of the prelimbic medial prefrontal cortex (PL) in the regulation of anxiety-like behaviors by pharmacologically activating the terminals of neuronal inputs or postsynaptic efferent neurons with a sodium channel activator veratrine. The extracellular glutamate levels were measured by in vivo microdialysis, and the behaviors were assessed with the open field (OF) test in mice simultaneously. The samples were collected every 10 min for 60 min, as basal levels of glutamate. The medium containing drugs were perfused for 30 min. The OF test was performed in the last 10 min of drug perfusion. After the drug treatments, the perfusion medium containing drugs was switched back to perfusion medium without drugs, and then samples were collected for another 90 min. The extracellular glutamate levels were significantly elevated after local perfusion of veratrine in the PL. At the same time, perfusion of veratrine in the PL produced anxiety-like behaviors in mice. Local coperfusion of a sodium channel blocker, lamotrigine, completely diminished the veratrine-induced elevated extracellular glutamate levels and the behavioral changes. Local coperfusion of an NMDA receptor antagonist, MK-801, but not a non-NMDA (AMPA/kainate) receptor antagonist, CNQX, completely diminished the behavioral changes without any effects on the veratrine-induced elevated extracellular glutamate levels. This study demonstrates that the activation of the PL with veratrine induces anxiety-like behaviors via NMDA receptor-mediated glutamatergic neurotransmission in mice. © 2014 Wiley Periodicals, Inc.

Expression of Bacillus protease (Protease BYA) from Bacillus sp. Y in Bacillus subtilis and enhancement of its specific activity by site-directed mutagenesis-improvement in productivity of detergent enzyme-.

PubMed

Tobe, Seiichi; Shimogaki, Hisao; Ohdera, Motoyasu; Asai, Yoshio; Oba, Kenkichi; Iwama, Masanori; Irie, Masachika

2006-01-01

An attempt was made to express protease BYA produced by an alkalophilic Bacillus sp. Y in Bacillus subtilis by gene engineering methods. The gene encoding protease BYA was cloned from Bacillus sp. Y, and expression vector pTA71 was constructed from the amylase promoter of Bacillus licheniformis, DNA fragments encoding the open reading frame of protease BYA, and pUB110. Protease BYA was secreted at an activity level of 5100 APU/ml in the common industrial culture medium of Bacillus subtilis transformed with pTA71. We then attempted to increase the specific activity of protease BYA by site-directed mutagenesis. Amino acid residue Ala29 next to catalytic Asp30 was replaced by one of three uncharged amino acid residues (Val29, Leu29, Ile29), and each mutant enzyme was expressed and isolated from the culture medium. Val29 mutant enzyme was secreted at an activity level of greater than 7000 APU/ml in culture medium, and its specific activity was 1.5-fold higher than that of the wild-type enzyme. Other mutant enzymes had specific activity similar to that of the original one and were less stabile than the wild-type enzyme. It can be thought that the substitution at amino acid residue 29 affects the level of activity and stability of protease BYA.

Antioxidant effect of mono- and dihydroxyphenols in sunflower oil with different levels of naturally present tocopherols

PubMed Central

Hrádková, Iveta; Merkl, Roman; Šmidrkal, Jan; Kyselka, Jan; Filip, Vladimír

2013-01-01

Antioxidant properties of mono- and dihydroxyphenolic acids and their alkyl esters were examined, with emphasis on the relationship between their molecular structure and antioxidant activity. Test media with different tocopherol level were used for determining the oxidative stability: original refined sunflower oil (total tocopherols 149.0 mg/kg), partially tocopherol-stripped sunflower oil (total tocopherols 8.7 mg/kg) and distilled fatty acid methyl esters (FAME) as a tocopherol-free medium. The chemical reaction of tocopherols with diazomethane tested for the purpose to eliminate their antioxidant activity failed due to the negligible degree of methylation of hydroxyl group in the tocopherol molecule. Caffeic acid and protocatechuic acid (3,4-dihydroxyphenolic acids) and their alkyl esters were found to be more active antioxidants than monohydroxyphenolic acid (p-hydroxybenzoic acid), 2,5-dihydroxyphenolic acid (gentisic acid), 3-methoxy-4-hydroxyphenolic acids (vanillic and ferulic acids) and their corresponding alkyl esters. Naturally present tocopherols in refined sunflower oil proved to have a synergistic effect on gentisic acid but not on its alkyl esters. In contrast, tocopherols showed an antagonistic effect on alkyl esters of caffeic acid, because their protection factors decreased with increasing level of tocopherols in the test medium. Moreover, the antioxidant activity of these alkyl esters decreased with increasing length of their alkyl chain in conformity with the polar paradox hypothesis. Practical applications: Tocopherols as naturally present antioxidants influence considerably the antioxidant activity of other antioxidants added to plant oils used as a test medium. Distilled fatty acid methyl esters prepared from refined sunflower oil may serve as an optimal tocopherol-free test medium. Some alkyl esters of phenolic acids were evaluated to be applicable as natural more lipophilic antioxidants in comparison with phenolic acids. PMID:23997655

Optical activity via Kerr nonlinearity in a spinning chiral medium

NASA Astrophysics Data System (ADS)

Khan, Anwar Ali; Bacha, Bakht Amin; Khan, Rahmat Ali

2016-11-01

Optical activity is investigated in a chiral medium by employing the four level cascade atomic model, in which the optical responses of the atomic medium are studied with Kerr nonlinearity. Light entering into a chiral medium splits into circular birefringent beams. The angle of divergence between the circular birefringent beams and the polarization states of the two light beams is manipulated with Kerr nonlinearity. In the stationary chiral medium the angle of divergence between the circular birefringent beams is calculated to be 1.3 radian. Furthermore, circular birefringence is optically controlled in a spinning chiral medium, where the maximum rotary photon drag angle for left (right) circularly polarized beam is ±1.1 (±1.5) microradian. The change in the angle of divergence between circular birefringent beams by rotary photon drag is calculated to be 0.4 microradian. The numerical results may help to understand image designing, image coding, discovery of photonic crystals and optical sensing technology.

Tween 80 effect on glucosyltransferase synthesis by Streptococcus salivarius.

PubMed Central

Wittenberger, C L; Beaman, A J; Lee, L N

1978-01-01

Streptococcus salivarius (ATCC 25975) produced very low or nondetectable amounts of the extracellular enzyme glucosyltransferase (GTase) when grown in a chemically defined medium. The addition of Tween 80 to this medium resulted in the production of markedly enhanced levels of the enzyme. Oleic acid, the methyl ester of oleic acid, and sucrose each could not substitute for Tween 80 in this regard. The surfactant had no direct activating effect on performed enzyme activity. Tween 80 also stimulated the production of GTase by concentrated cells suspended in defined medium during a time when no measurable growth occurred. Under these conditions, the stimulatory effect of Tween 80 was blocked by chloramphenicol. It was further found that the surfactant dramatically stimulated the differential rate of GTase synthesis. These and other data strongly suggest that Tween 80 stimulates the production of extracellular GTase by acting either directly or indirectly at the level of enzyme synthesis. PMID:618839

Production of L-asparaginase by filamentous fungi.

PubMed

Sarquis, Maria Inez de Moura; Oliveira, Edna Maria Morais; Santos, Alberdan Silva; Costa, Gisela Lara da

2004-08-01

L-asparaginase production was investigated in the filamentous fungi Aspergillus tamarii and Aspergillus terreus. The fungi were cultivated in medium containing different nitrogen sources. A. terreus showed the highest L-asparaginase (activity) production level (58 U/L) when cultivated in a 2% proline medium. Both fungi presented the lowest level of L-asparaginase production in the presence of glutamine and urea as nitrogen sources. These results suggest that L-asparaginase production by of filamentous fungi is under nitrogen regulation.

Optimization of end-pumped, actively Q-switched quasi-III-level lasers.

PubMed

Jabczynski, Jan K; Gorajek, Lukasz; Kwiatkowski, Jacek; Kaskow, Mateusz; Zendzian, Waldemar

2011-08-15

The new model of end-pumped quasi-III-level laser considering transient pumping processes, ground-state-depletion and up-conversion effects was developed. The model consists of two parts: pumping stage and Q-switched part, which can be separated in a case of active Q-switching regime. For pumping stage the semi-analytical model was developed, enabling the calculations for final occupation of upper laser level for given pump power and duration, spatial profile of pump beam, length and dopant level of gain medium. For quasi-stationary inversion, the optimization procedure of Q-switching regime based on Lagrange multiplier technique was developed. The new approach for optimization of CW regime of quasi-three-level lasers was developed to optimize the Q-switched lasers operating with high repetition rates. Both methods of optimizations enable calculation of optimal absorbance of gain medium and output losses for given pump rate. © 2011 Optical Society of America

Effect of Lipid Peroxidation Products on the Activity of Human Retinol Dehydrogenase 12 (RDH12) and Retinoid Metabolism

PubMed Central

Lee, Seung-Ah; Belyaeva, Olga V.; Kedishvili, Natalia Y.

2008-01-01

SUMMARY Mutations in human Retinol Dehydrogenase 12 (RDH12) are known to cause photoreceptor cell death but the physiological function of RDH12 in photoreceptors remains poorly understood. In vitro, RDH12 recognizes both retinoids and medium-chain aldehydes as substrates. Our previous study suggested that RDH12 protects cells against toxic levels of retinaldehyde and retinoic acid [Lee et al., J. Biol. Chem. 282 (2007) 35621–35628]. Here, we investigated whether RDH12 can also protect cells against highly reactive medium-chain aldehydes. Analysis of cell survival demonstrated that RDH12 was protective against nonanal but not against 4-hydroxynonenal. At high concentrations, nonanal inhibited the activity of RDH12 towards retinaldehyde, suggesting that nonanal was metabolized by RDH12. 4-Hydroxynonenal did not inhibit the RDH12 retinaldehyde reductase activity, but it strongly inhibited the activities of lecithin:retinol acyl transferase and aldehyde dehydrogenase, resulting in decreased levels of retinyl esters and retinoic acid and accumulation of unesterified retinol. Thus, the results of this study showed that RDH12 is more effective in protection against retinaldehyde than against medium-chain aldehydes, and that medium-chain aldehydes, especially 4-hydroxynonenal, severely disrupt cellular retinoid homeostasis. Together, these findings provide a new insight into the effects of lipid peroxidation products and the impact of oxidative stress on retinoid metabolism. PMID:18396173

[Change in concentration of cytosolic Ca2+ caused by extracellular ATP and ecto-ATP-ase activity in thymocytes and transformed MT-4 cells].

PubMed

Hrebinyk, S M; Artemenko, O Iu; Hryniuk, I I; Perepelitsyna, O M; Matyshevs'ka, O P

2009-01-01

The comparative study of extracellular ATP (ATP0) effect on free cytosolic calcium concentration ([Ca2+]i) in normal (isolated rat thymocytes) and transformed (leukosis MT-4 line) T-cells was carried out. Addition of 1 mM ATP to Ca-free incubation medium of both types of cells, loaded with indo-1, had no effect on [Ca2+]i level. Upon subsequent addition of 1 mM CaCl2 to the incubation medium the rapid and significant increase of [Ca2+]i in MT-4 cells was registered. This effect was maintained within 10 min and was not inhibited by phospholipase C inhibitor 0.2 mM neomycin, that was induced by cation entry into the cells from the extracellular medium. Both types of cells were shown to demonstrate ecto-ATPase activity in the presence of 1 mM MgCl2 or CaC12 in the incubation medium. Estimation of kinetic parameters has indicated that the maximum rate of extracellular ATP hydrolysis by MT-4 cells is higher and Mg2+ and Ca2+ activation constants are lower as compared to respective parameters of ATP hydrolysis by thymocytes. The possible functional significance of the increased level of ecto-ATPase activity in malignantly transformed cells is discussed.

Use of care management practices in small- and medium-sized physician groups: do public reporting of physician quality and financial incentives matter?

PubMed

Alexander, Jeffrey A; Maeng, Daniel; Casalino, Lawrence P; Rittenhouse, Diane

2013-04-01

To examine the effect of public reporting (PR) and financial incentives tied to quality performance on the use of care management practices (CMPs) among small- and medium-sized physician groups. Survey data from The National Study of Small and Medium-sized Physician Practices were used. Primary data collection was also conducted to assess community-level PR activities. The final sample included 643 practices engaged in quality reporting; about half of these practices were subject to PR. We used a treatment effects model. The instrumental variables were the community-level variables that capture the level of PR activity in each community in which the practices operate. (1) PR is associated with increased use of CMPs, but the estimate is not statistically significant; (2) financial incentives are associated with greater use of CMPs; (3) practices' awareness/sensitivity to quality reports is positively related to their use of CMPs; and (4) combined PR and financial incentives jointly affect CMP use to a greater degree than either of these factors alone. Small- to medium-sized practices appear to respond to PR and financial incentives by greater use of CMPs. Future research needs to investigate the appropriate mix and type of incentive arrangements and quality reporting. © Health Research and Educational Trust.

Sialidase activities of cultured human fibroblasts and the metabolism of GM3 ganglioside

DOE Office of Scientific and Technical Information (OSTI.GOV)

Usuki, S.; Lyu, S.C.; Sweeley, C.C.

1988-05-15

Free sialic acid has been found in the cell-conditioned medium of human foreskin fibroblasts. It is proposed that the accumulation of extracellular sialic acid may result from the hydrolysis of GM3 ganglioside on the cell surface of these fibroblasts. Sialidase activities with GM3 ganglioside and sialyllactitol as substrates were demonstrated in cell-conditioned medium, and the levels of their activities correlated positively with cell density. The GM3 sialidase activity at pH 4.5 was 4.1 and 38 pmol/h/ml of medium at sparse and confluent densities, respectively; the corresponding activities with sialyllactitol as the substrate were 12 and 75 pmol/h/ml of medium (pHmore » 4.5). The pH versus activity profiles with GM3 as the substrate suggested the presence of a second sialidase with an optimal activity at pH 6.5 in the conditioned medium of preconfluent cells. This activity was virtually absent in the medium of contact-inhibited cells and could not be assayed with sialyllactitol as the substrate. The turnover of cell surface GM3 was assessed by pulse labeling human foreskin fibroblasts with a radioactive precursor of sialic acid ((1-14C)N-acetylmannosamine) and a radioactive precursor of ceramide ((3,3-3H2)serine). During a chase period of 24 h turnover of the doubly labeled cellular GM3 was observed; there was a loss of about 35% of the 14C-labeled sialic acid without any measureable loss of 3H-labeled ceramide from GM3. We have speculated that the enzyme-catalyzed removal of sialic acid from the GM3 ganglioside on the extracellular aspect of the plasma membrane may be a necessary event involved in the modulation of cell growth.« less

Relationship of weight status, physical activity and screen time with academic achievement in adolescents.

PubMed

García-Hermoso, Antonio; Marina, Raquel

The aim of this study was to examine the relationship of weight status, physical activity and screen time with academic achievement in Chilean adolescents. The present cross-sectional study included 395 adolescents. The International Obesity Task Force cut-off points were used to define the weight status. Physical activity was assessed using the Physical Activity Questionnaire for Adolescents and screen time was assessed using several questions about television, videogame and computer use. Academic achievement was measured using the mean of the grades obtained in mathematics and language subjects. In both genders, adolescents with obesity and excessive screen time earned worse grades compared to their non-obese peers and their peers that complied with screen time recommendations. The logistic regression analysis showed that adolescents with obesity, classified with medium-low physical activity and excessive screen time recommendations (excess ≥2h/day) are less likely to obtain high academic achievement (boys: OR=0.26; girls: OR=0.23) compared to their non-obese peers, high levels of physical activity and those who comply with the current screen time recommendations. Similar results were observed in adolescents with obesity and classified with medium-low physical activity (boys: OR=0.46; girls: OR=0.33) or excessive screen time (boys: OR=0.35; girls: OR=0.36) compared to adolescents with high levels of physical activity and those who complied with the screen time recommendations, respectively. This study shows that when combined, obesity, low-medium levels of physical activity and excessive screen time might be related to poor academic achievement. Copyright © 2015 Asia Oceania Association for the Study of Obesity. Published by Elsevier Ltd. All rights reserved.

Breast carcinoma cells modulate the chemoattractive activity of human bone marrow-derived mesenchymal stromal cells by interfering with CXCL12.

PubMed

Wobus, Manja; List, Catrin; Dittrich, Tobias; Dhawan, Abhishek; Duryagina, Regina; Arabanian, Laleh S; Kast, Karin; Wimberger, Pauline; Stiehler, Maik; Hofbauer, Lorenz C; Jakob, Franz; Ehninger, Gerhard; Anastassiadis, Konstantinos; Bornhäuser, Martin

2015-01-01

We investigated whether breast tumor cells can modulate the function of mesenchymal stromal cells (MSCs) with a special emphasis on their chemoattractive activity towards hematopoietic stem and progenitor cells (HSPCs). Primary MSCs as well as a MSC line (SCP-1) were cocultured with primary breast cancer cells, MCF-7, MDA-MB231 breast carcinoma or MCF-10A non-malignant breast epithelial cells or their conditioned medium. In addition, the frequency of circulating clonogenic hematopoietic progenitors was determined in 78 patients with breast cancer and compared with healthy controls. Gene expression analysis of SCP-1 cells cultured with MCF-7 medium revealed CXCL12 (SDF-1) as one of the most significantly downregulated genes. Supernatant from both MCF-7 and MDA-MB231 reduced the CXCL12 promoter activity in SCP-1 cells to 77% and 47%, respectively. Moreover, the CXCL12 mRNA and protein levels were significantly reduced. As functional consequence of lower CXCL12 levels, we detected a decreased trans-well migration of HSPCs towards MSC/tumor cell cocultures or conditioned medium. The specificity of this effect was confirmed by blocking studies with the CXCR4 antagonist AMD3100. Downregulation of SP1 and increased miR-23a levels in MSCs after contact with tumor cell medium as well as enhanced TGFβ1 expression were identified as potential molecular regulators of CXCL12 activity in MSCs. Moreover, we observed a significantly higher frequency of circulating colony-forming hematopoietic progenitors in patients with breast cancer compared with healthy controls. Our in vitro results propose a potential new mechanism by which disseminated tumor cells in the bone marrow may interfere with hematopoiesis by modulating CXCL12 in protected niches. © 2014 UICC.

A Study of the Diffusion, Electrochemical Mobility and Removal of Dissolved Copper in a Saturated Porous Medium.

DTIC Science & Technology

1980-11-01

aqueous solutions : use of activity coefficients in transition-state models: Geochimica et Cosmochimica Acta, v...native state at 25°C at any activity level below 10+46.0 in an aqueous solution . Because such an activity level is impossible, sodium cannot be reduced...stoichiometric coefficients . It is necessary to calculqte the activity coefficients of dissolved copper in the test solutions in order to render an

[Degradation of anthraquinone blue by Trametes trogii].

PubMed

Levin, L; Jordan, A; Forchiassin, F; Viale, A

2001-01-01

The ability of the white rot fungus Trametes trogii BAFC 463 (high producer of ligninolytic enzymes, especially laccase and manganese peroxidase) to degrade the dye anthraquinone blue, refractory to bacterial attack, was evaluated. Both tropho- and idiophasic T. trogii cultures in synthetic medium (glucose/asparagine) and complex medium (malt extract/glucose) were able to transform up to 88% dye in 4 hours. The activity of laccase, an oxygen-dependent phenoloxidase which was present at high levels in all the conditions assayed, might be related to the ability of the fungus to degrade the colorant. This is supported by the fact that in bioreactor experiences carried out at pH 4.5 the addition of anthraquinone blue caused a decrease in the levels of soluble oxygen. However, although high levels of laccase were produced at pH 7.5, the enzyme was not active, and neither dye transformation nor loss in the levels of soluble oxygen were quantified.

Hepatoprotective activity of sea cucumber Phyllophorus sp. extract in carp (Cyprinus carpio)

NASA Astrophysics Data System (ADS)

Sulmartiwi, Laksmi; Triastuti, Juni; Andriyono, Sapto; Umami, Mardiah Rahma

2017-02-01

Many procedures continuously in aquaculture and scientific research like tagging and vaccinating cause pain, involving damaging tissue and also cause stress responses in fish. Stress responses in fish influence liver because liver have vital role to supply energy and metabolism. Histology alteration in liver is caused by stress response like changes of vacuolation hepatocyte and characteristic colour. Triterpenoid was known had hepatoprotective activity. One of marine organism contained triterpenoid was sea cucumber. Result of research showed that liver tissue in fish with injected acetic acid 5 % (in upper lip) as pain stimulus have histopathology damages such as pyknosis (medium damage level) and oedema (heavy damage level) after 8 hour injection. Injected Lidocaine 1mg/fish as analgesic drug have histopathology damages such as oedema (heavy damages level), necrosis and pyknosis (low damages level). Injected acetic acid 5 % (in upper lip) and ethanolic extract of sea cucumber Phyllophorus sp. dose 5 mg/50 gr body weight shown histopathology damages such as necrosis, edema (medium damage level) and pyknosis (low damage level).

Chloride-dependency of amyloid beta protein-induced enhancement of glutamate neurotoxicity in cultured rat hippocampal neurons.

PubMed

Zhang, Nan-Yan; Kitagawa, Kaori; Wu, Bo; Xiong, Zheng-Mei; Otani, Hitomi; Inagaki, Chiyoko

2006-05-15

In our previous studies, pathophysiological concentrations of amyloid-beta (Abeta) proteins increased intracellular Cl(-) concentration ([Cl(-)]i) and enhanced glutamate neurotoxicity in primary cultured neurons, suggesting Cl(-)-dependent changes in glutamate signaling. To test this possibility, we examined the effects of isethionate-replaced low Cl(-) medium on the Abeta-induced enhancement of glutamate neurotoxicity in the primary cultured rat hippocampal neurons. In a normal Cl(-) (135 mM) medium, treatment with 10 nM Abeta25-35 for 2 days increased neuronal [Cl(-)]i to a level three times higher than that of control as assayed using a Cl(-)-sensitive fluorescent dye, while in a low Cl(-) (16 mM) medium such an Abeta25-35-induced increase in [Cl(-)]i was not observed. The Abeta treatment aggravated glutamate neurotoxicity in a normal Cl(-) medium as measured by mitochondrial reducing activity and lactate dehydrogenase (LDH) release, while in a low Cl(-) medium the Abeta treatment did not enhance glutamate toxicity. Upon such Abeta plus glutamate treatment under a normal Cl(-) condition, activated anti-apoptotic molecule Akt (Akt-pS473) level monitored by Western blot significantly decreased to 74% of control. Under a low Cl(-) condition, a resting Akt-pS473 level was higher than that under a normal Cl(-) condition and did not significantly change upon Abeta plus glutamate treatment. Tyrosine phosphorylation levels of 110 and 60 kDa proteins (pp110 and pp60) increased upon Abeta plus glutamate treatment under a normal Cl(-), but not low Cl(-), condition. These findings indicated that Abeta-induced enhancement of glutamate neurotoxicity is Cl(-)-dependent. Chloride-sensitive Akt pathway and tyrosine phosphorylation of proteins (pp110 and pp60) may be involved in this process.

75 FR 61432 - Takes of Marine Mammals Incidental to Specified Activities; Construction of the Parsons Slough...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-10-05

... a medium, such as air or water. Sound levels are compared to a reference sound pressure to identify the medium. For air and water, these reference pressures are ``re 20 microPa'' and ``re 1 microPa... produce a wide range of social signals, most occurring at relatively low frequencies (Southall et al...

Effects of melanin on the accumulation of exopolysaccharides by Aureobasidium pullulans grown on nitrate.

PubMed

Zheng, Weifa; Campbell, Bradley S; McDougall, Barbara M; Seviour, Robert J

2008-11-01

Aureobasidium pullulans produced pullulan and melanin when grown in medium containing low nitrate levels. With high nitrate concentrations, however, this fungus produced a mixture of exopolysaccharides (EPS) without melanin synthesis. At 0.78 g l(-1) N as nitrate, where no melanin synthesis occurred, maximum EPS yields reached 6.92 g l(-1) and then decreased to the final yield of 2.36 g l(-1). Following melanin addition (0.1 g l(-1)), yields reached 7.02 g l(-1) at 48 h and fell to a final yield of 5.21 g l(-1). The EPS produced in high nitrate medium contained both pullulan and (1-->3)-beta-glucan, but only pullulan was produced with melanin-supplementation. With melanin addition a doubling of (1-->3)-beta-glucanase activity was observed in high nitrate medium compared to that without supplementation. On the other hand amylolytic activities disappeared in medium with melanin production or addition. Culture filtrates sustained a higher reducing capacity (RC) when melanin was present. Low RC appeared to reduce (1-->3)-beta-glucanase activity and increase amylolytic activities. Thus, higher RC appears to inhibit production/activity of amylose-degrading enzymes capable of degrading pullulan, and stimulates (1-->3)-beta-glucanase synthesis/activity, leading to a preferential accumulation of pullulan.

Production, secretion, and stability of human secreted alkaline phosphatase in tobacco NT1 cell suspension cultures.

PubMed

Becerra-Arteaga, Alejandro; Mason, Hugh S; Shuler, Michael L

2006-01-01

Tobacco NT1 cell suspension cultures secreting active human secreted alkaline phosphatase (SEAP) were generated for the first time as a model system to study recombinant protein production, secretion, and stability in plant cell cultures. The SEAP gene encodes a secreted form of the human placental alkaline phosphatase (PLAP). During batch culture, the highest level of active SEAP in the culture medium (0.4 U/mL, corresponding to approximately 27 mg/L) was observed at the end of the exponential growth phase. Although the level of active SEAP decreased during the stationary phase, the activity loss did not appear to be due to SEAP degradation (based on Western blots) but due to SEAP denaturation. The protein-stabilizing agents polyvinylpirrolidone (PVP) and bacitracin were added extracellularly to test for their ability to reduce the loss of SEAP activity during the stationary phase. Bacitracin (100 mg/L) was the most effective treatment at sustaining activity levels for up to 17 days post-subculture. Commercially available human placental alkaline phosphatase (PLAP) was used to probe the mechanism of SEAP deactivation. Experiments with PLAP in sterile and conditioned medium corroborated the denaturation of SEAP by factors generated by cell growth and not due to simple proteolysis. We also show for the first time that the factors promoting activity loss are heat labile at 95 degrees C but not at 70 degrees C, and they are not inactivated after a 5 day incubation period under normal culture conditions (27 degrees C). In addition, there were no significant changes in pH or redox potential when comparing sterile and cell-free conditioned medium during PLAP incubation, indicating that these factors were unimportant.

PHYSICAL EFFECTS OCCURRING DURING GENERATION AND AMPLIFICATION OF LASER RADIATION: Discharge energy balance in the nitrogen-containing active medium of an electron-beam-controlled CO laser

NASA Astrophysics Data System (ADS)

Dolinina, V. I.; Koterov, V. N.; Pyatakhin, Mikhail V.; Urin, B. M.

1989-02-01

Numerical methods were used to investigate theoretically the dynamics of the energy balance of a discharge in a CO-N2 mixture, taking into account the mutual influence of the distributions of the electron energy and of the populations of the molecules over the vibrational levels. It was shown that this influence plays a decisive part in substantially redistributing the pump energy between the vibrational levels of the CO and N2 molecules in favor of the N2 molecules. A stabilizing action of the nitrogen on the thermal regime of the CO laser-active medium was discovered and the range of optimal CO:N2 ratios was determined.

Effect of blanching treatments on antioxidant activity of frozen green capsicum (Capsicum annuum L. var bell pepper) using radical scavenging activity (DPPH) assay

NASA Astrophysics Data System (ADS)

Azizzuddin, Norafida; Abdullah, Aminah

2016-11-01

Blanching treatments are needed to deactivate enzymes in frozen vegetables. Antioxidant activity using DPPH radical scavenging activity assay were evaluated in steaming, boiling water, and microwave blanching at different temperature, time and microwave power level on frozen green capsicum. Green capsicum was chosen for frozen treatment compared to other capsicum with different maturity index because of the firm texture. The objective of this study was to compare the antioxidant activity of frozen green capsicum between conventional and Oxi Count Kit® assay for DPPH radical scavenging activity. Results showed frozen green capsicum blanched using microwave at high level/90 seconds (sample J) contained higher level of DPPH in both conventional method and Oxi Count Kit® compared to other treatments. However, there were no significant differences between sample J and fresh sample (sample A). Overall, the sequences from highest to lowest in blanching treatments for both DPPH conventional method, and DPPH Oxi Count Kit® were J (microwave high level/90 seconds) > A (Fresh) > H (Microwave Medium Level/120 seconds) > D (Boiling Water 80°C/150 seconds) > K (Microwave High Level/120 seconds) > I (Microwave Medium Level/150 seconds) > F (Microwave Low Level/150 seconds)> B (Steam 100°C/150 seconds) > E (Boiling Water 100°C /120 seconds) > G (Microwave Low Level /180 seconds)> C (Steam 100°C/180 seconds). Almost all frozen green capsicum samples showed no significant differences for comparison between test using DPPH conventional method and Oxi Count Kit®. Frozen storage for 0, and 3rd months showed no significant differences which indicate no changes on antioxidant activity during frozen storage at -18°C.

CATALASE AND SUPEROXIDE DISMUTASE OF ROOT-COLONIZING SAPROPHYTIC FLUORESCENT PSEUDOMONADS

EPA Science Inventory

Root-colonizing, saprophytic fluorescent pseudomonads of the Pseudomonas putida-P. fluorescens group express similar levels of catalase and superoxide dismutase activities during growth on a sucrose- and amino acid-rich medium. ncreased specific activities of catalase but not sup...

Protective properties of ginsenoside Rb1 against UV-B radiation-induced oxidative stress in human dermal keratinocytes.

PubMed

Oh, Sun-Joo; Kim, Kyunghoon; Lim, Chang-Jin

2015-06-01

Ginsenosides, also known as ginseng saponins, are responsible for most pharmacological effect of ginseng. Ginsenoside Rb1 (Rb1) exerts a variety of pharmacological properties, including anti-inflammatory, antistress, anti-aging and anti-neurodegenerative activities. The aim of the present work was to assess the skin anti-photoaging properties of Rb1 in human dermal keratinocyte HaCaT cells. The anti-photoaging activity was evaluated by analyzing the levels of reactive oxygen species (ROS) and matrix metalloproteinases (MMPs) as well as cell viability for HaCaT cells under UV-B irradiation. Rb1 was able to suppress the ROS levels which were elevated under UV-B irradiation, and unable to influence cellular survival in UV-B-irradiated HaCaT cells. Rb1 diminished the enhancement of MMP-2 gelatinolytic activity in conditioned medium, which corresponded with the decreased MMP-2 protein levels in both conditioned medium and cellular lysate prepared from UV-B-irradiated HaCaT cultures. Rb1 could restore the total glutathione (GSH) and superoxide dismutase (SOD) activity diminished in UV-B-irradiated HaCaT cells. Ginsenoside Rb1 possesses skin anti-photoaging properties through scavenging ROS and decreasing MMP-2 levels possibly by enhancing antioxidant activity in keratinocytes under UV-B irradiation.

Screening of Actinomycetes from mangrove ecosystem for L-asparaginase activity and optimization by response surface methodology.

PubMed

Usha, Rajamanickam; Mala, Krishnaswami Kanjana; Venil, Chidambaram Kulandaisamy; Palaniswamy, Muthusamy

2011-01-01

Marine actinomycetes were isolated from sediment samples collected from Pitchavaram mangrove ecosystem situated along the southeast coast of India. Maximum actinomycete population was noted in rhizosphere region. About 38% of the isolates produced L-asparaginase. One potential strain KUA106 produced higher level of enzyme using tryptone glucose yeast extract medium. Based on the studied phenotypic characteristics, strain KUA106 was identified as Streptomyces parvulus KUA106. The optimization method that combines the Plackett-Burman design, a factorial design and the response surface method, which were used to optimize the medium for the production of L-asparaginase by Streptomycetes parvulus. Four medium factors were screened from eleven medium factors by Plackett-Burman design experiments and subsequent optimization process to find out the optimum values of the selected parameters using central composite design was performed. Asparagine, tryptone, d) extrose and NaCl components were found to be the best medium for the L-asparaginase production. The combined optimization method described here is the effective method for screening medium factors as well as determining their optimum level for the production of L-asparaginase by Streptomycetes parvulus KUAP106.

Human periodontal ligament stem cells secretome from multiple sclerosis patients suppresses NALP3 inflammasome activation in experimental autoimmune encephalomyelitis

PubMed Central

Soundara Rajan, Thangavelu; Giacoppo, Sabrina; Diomede, Francesca; Bramanti, Placido; Trubiani, Oriana; Mazzon, Emanuela

2017-01-01

Research in recent years has largely explored the immunomodulatory effects of mesenchymal stem cells (MSCs) and their secretory products, called “secretome,” in the treatment of neuroinflammatory diseases. Here, we examined whether such immunosuppressive effects might be elicited due to inflammasome inactivation. To this end, we treated experimental autoimmune encephalomyelitis (EAE) mice model of multiple sclerosis (MS) with the conditioned medium or purified exosomes/microvesicles (EMVs) obtained from relapsing-remitting-MS patients human periodontal ligament stem cells (hPDLSCs) and investigated the regulation of NALP3 inflammasome. We noticed enhanced expression of NALP3, Cleaved Caspase 1, interleukin (IL)-1β, and IL-18 in EAE mouse spinal cord. Conversely, hPDLSCs-conditioned medium and EMVs significantly blocked NALP3 inflammasome activation and provided protection from EAE. Reduction in NALP3, Cleaved Caspase 1, IL-1β, and IL-18 level was noticed in conditioned medium and EMVs-treated EAE mice. Pro-inflammatory Toll-like receptor (TLR)-4 and nuclear factor (NF)-κB were elevated in EAE, while hPDLSCs-conditioned medium and EMVs treatment reduced their expression and increased IκB-α expression. Characterization of hPDLSCs-conditioned medium showed substantial level of anti-inflammatory IL-10, transforming growth factor (TGF)-β, and stromal cell–derived factor 1α (SDF-1α). We propose that the immunosuppressive role of hPDLSCs-derived conditioned medium and EMVs in EAE mice may partly attribute to the presence of soluble immunomodulatory factors, NALP3 inflammasome inactivation, and NF-κB reduction. PMID:28764573

Strain selection and medium optimization for glucoamylase production from industrial potato waste by Aspergillus niger.

PubMed

Izmirlioglu, Gulten; Demirci, Ali

2016-06-01

Glucoamylase is one of the most common enzymes used in the food industry to break down starch into its monomers. Glucoamylase production and its activity are highly dependent on medium composition. Starch is well known as a glucoamylase inducer, and utilization of industrial starchy potato waste is an inexpensive way of improving glucoamylase production. Since glucoamylase production is highly dependent on medium composition, in this study medium optimization for glucoamylase production was considered to enhance glucoamylase activity. Among the evaluated microbial species, Aspergillus niger van Tieghem was found to be the best glucoamylase-producing fungus. The Plackett-Burman design was used to screen various medium ingredients, and malt extract, FeSO4 .7H2 O and CaCl2 ·2H2 O were found to have significant effects on glucoamylase production. Finally, malt extract, FeSO4 .7H2 O and CaCl2 .2H2 O were optimized by using a central composite design of response surface methodology. The results showed that the optimal medium composition for A. niger van Tieghem was 50 g L(-1) industrial waste potato mash supplemented with 51.82 g L(-1) malt extract, 9.27 g L(-1) CaCl2 ·2H2 O and 0.50 g L(-1) FeSO4 .7H2 O. At the end of optimization, glucoamylase activity and glucose production were improved 126% and 98% compared to only industrial waste potato mash basal medium; 274.4 U mL(-1) glucoamylase activity and 41.7 g L(-1) glucose levels were achieved, respectively. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

Reduction in voltage-gated K+ channel activity in primary sensory neurons in painful diabetic neuropathy: role of brain-derived neurotrophic factor.

PubMed

Cao, Xue-Hong; Byun, Hee-Sun; Chen, Shao-Rui; Cai, You-Qing; Pan, Hui-Lin

2010-09-01

Abnormal hyperexcitability of primary sensory neurons plays an important role in neuropathic pain. Voltage-gated potassium (Kv) channels regulate neuronal excitability by affecting the resting membrane potential and influencing the repolarization and frequency of the action potential. In this study, we determined changes in Kv channels in dorsal root ganglion (DRG) neurons in a rat model of diabetic neuropathic pain. The densities of total Kv, A-type (IA) and sustained delayed (IK) currents were markedly reduced in medium- and large-, but not in small-, diameter DRG neurons in diabetic rats. Quantitative RT-PCR analysis revealed that the mRNA levels of IA subunits, including Kv1.4, Kv3.4, Kv4.2, and Kv4.3, in the DRG were reduced approximately 50% in diabetic rats compared with those in control rats. However, there were no significant differences in the mRNA levels of IK subunits (Kv1.1, Kv1.2, Kv2.1, and Kv2.2) in the DRG between the two groups. Incubation with brain-derived neurotrophic factor (BDNF) caused a large reduction in Kv currents, especially IA currents, in medium and large DRG neurons from control rats. Furthermore, the reductions in Kv currents and mRNA levels of IA subunits in diabetic rats were normalized by pre-treatment with anti-BDNF antibody or K252a, a TrkB tyrosine kinase inhibitor. In addition, the number of medium and large DRG neurons with BDNF immunoreactivity was greater in diabetic than control rats. Collectively, our findings suggest that diabetes primarily reduces Kv channel activity in medium and large DRG neurons. Increased BDNF activity in these neurons likely contributes to the reduction in Kv channel function through TrkB receptor stimulation in painful diabetic neuropathy.

Medium-chain fatty acids as ligands for orphan G protein-coupled receptor GPR84.

PubMed

Wang, Jinghong; Wu, Xiaosu; Simonavicius, Nicole; Tian, Hui; Ling, Lei

2006-11-10

Free fatty acids (FFAs) play important physiological roles in many tissues as an energy source and as signaling molecules in various cellular processes. Elevated levels of circulating FFAs are associated with obesity, dyslipidemia, and diabetes. Here we show that GPR84, a previously orphan G protein-coupled receptor, functions as a receptor for medium-chain FFAs with carbon chain lengths of 9-14. Medium-chain FFAs elicit calcium mobilization, inhibit 3',5'-cyclic AMP production, and stimulate [35S]guanosine 5'-O-(3-thiotriphosphate) binding in a GPR84-dependent manner. The activation of GPR84 by medium-chain FFAs couples primarily to a pertussis toxin-sensitive G(i/o) pathway. In addition, we show that GPR84 is selectively expressed in leukocytes and markedly induced in monocytes/macrophages upon activation by lipopolysaccharide. Furthermore, we demonstrate that medium-chain FFAs amplify lipopolysaccharide-stimulated production of the proinflammatory cytokine interleukin-12 p40 through GPR84. Our results indicate a role for GPR84 in directly linking fatty acid metabolism to immunological regulation.

CNTF-ACM promotes mitochondrial respiration and oxidative stress in cortical neurons through upregulating L-type calcium channel activity.

PubMed

Sun, Meiqun; Liu, Hongli; Xu, Huanbai; Wang, Hongtao; Wang, Xiaojing

2016-09-01

A specialized culture medium termed ciliary neurotrophic factor-treated astrocyte-conditioned medium (CNTF-ACM) allows investigators to assess the peripheral effects of CNTF-induced activated astrocytes upon cultured neurons. CNTF-ACM has been shown to upregulate neuronal L-type calcium channel current activity, which has been previously linked to changes in mitochondrial respiration and oxidative stress. Therefore, the aim of this study was to evaluate CNTF-ACM's effects upon mitochondrial respiration and oxidative stress in rat cortical neurons. Cortical neurons, CNTF-ACM, and untreated control astrocyte-conditioned medium (UC-ACM) were prepared from neonatal Sprague-Dawley rat cortical tissue. Neurons were cultured in either CNTF-ACM or UC-ACM for a 48-h period. Changes in the following parameters before and after treatment with the L-type calcium channel blocker isradipine were assessed: (i) intracellular calcium levels, (ii) mitochondrial membrane potential (ΔΨm), (iii) oxygen consumption rate (OCR) and adenosine triphosphate (ATP) formation, (iv) intracellular nitric oxide (NO) levels, (v) mitochondrial reactive oxygen species (ROS) production, and (vi) susceptibility to the mitochondrial complex I toxin rotenone. CNTF-ACM neurons displayed the following significant changes relative to UC-ACM neurons: (i) increased intracellular calcium levels (p < 0.05), (ii) elevation in ΔΨm (p < 0.05), (iii) increased OCR and ATP formation (p < 0.05), (iv) increased intracellular NO levels (p < 0.05), (v) increased mitochondrial ROS production (p < 0.05), and (vi) increased susceptibility to rotenone (p < 0.05). Treatment with isradipine was able to partially rescue these negative effects of CNTF-ACM (p < 0.05). CNTF-ACM promotes mitochondrial respiration and oxidative stress in cortical neurons through elevating L-type calcium channel activity.

Studies of the Acetate Kinase-Phosphotransacetylase and the Butanediol-Forming Systems in Aerobacter aerogenes

PubMed Central

Brown, T. D. K.; Pereira, C. R. S.; Størmer, F. C.

1972-01-01

Mutants of Aerobacter aerogenes devoid of acetate kinase and phosphotransacetylase activities were isolated by selection for resistance to fluoroacetate on lactate medium. The mutants were used to study the role of the acetate kinase-phosphotransacetylase system in growth on acetate and glucose. Acetate kinase-negative and phosphotransacetylase-negative mutants were unable to grow on acetate minimal medium. Their growth rates on glucose minimal medium were identical with that of the parent strain under aerobic conditions, but lower growth rates were observed in the mutant strains during anaerobic growth on glucose medium. The mutants were unable to incorporate [2-14C]-acetate rapidly while growing on glycerol. Variations in acetate kinase and phosphotransacetylase levels during growth on glucose were studied. The specific activities of the enzymes increased approximately fivefold during aerobic growth on glucose in batch culture. The enzyme levels were also studied during anaerobic growth on glucose at constant pH (pH 5.8 and 7.0). Smaller increases in specific activities were found under these conditions. The role of acetate in the induction of the diacetyl (acetoin) reductase was investigated using a mutant deficient in both acetate kinase and phosphotransacetylase. The effect of pH on the induction of this enzyme during growth on glucose under anaerobic conditions was tested. The data support the idea that free acetic acid is the inducer for the enzymes of the butanediol-forming pathway in A. aerogenes. PMID:4640502

Expression of the Sinorhizobium meliloti small RNA gene mmgR is controlled by the nitrogen source.

PubMed

Ceizel Borella, Germán; Lagares, Antonio; Valverde, Claudio

2016-05-01

Small non-coding regulatory RNAs (sRNAs) are key players in post-transcriptional regulation of gene expression. Hundreds of sRNAs have been identified in Sinorhizobium meliloti, but their biological function remains unknown for most of them. In this study, we characterized the expression pattern of the gene encoding the 77-nt sRNA MmgR in S. meliloti strain 2011. A chromosomal transcriptional reporter fusion (PmmgR-gfp) showed that the mmgR promoter is active along different stages of the interaction with alfalfa roots. In pure cultures, PmmgR-gfp activity paralleled the sRNA abundance indicating that mmgR expression is primarily controlled at the level of transcriptional initiation. PmmgR-gfp activity was higher during growth in rhizobial defined medium (RDM) than in TY medium. Furthermore, PmmgR-gfp was induced at 60 min after shifting growing cells from TY to RDM medium, i.e. shorter than the cell doubling time. In defined RDM medium containing NO3 (-), both PmmgR-gfp and MmgR level were repressed by the addition of tryptone or single amino acids, suggesting that mmgR expression depends on the cellular nitrogen (N) status. In silico analysis failed to detect conserved motifs upstream the promoter RNA polymerase binding site, but revealed a strongly conserved motif centered at -28 that may be linked to the observed regulatory pattern by the N source. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Modulation of CycD3;1-CDK complexes by phytohormones and sucrose during maize germination.

PubMed

Garza-Aguilar, Sara M; Lara-Núñez, Aurora; García-Ramírez, Elpidio; Vázquez-Ramos, Jorge M

2017-05-01

Maize CycD3;1 associates to CDKA or CDKB1;1 proteins during germination and the complexes formed develop kinase activity. These complexes appear to vary in size as germination proceeds, suggesting association to different sets of proteins. CycD3;1 and associated CDK proteins respond to phytohormones and sucrose. Results revealed a reduction in the CycD3;1 protein amount along germination in the presence of indoleacetic acid (IAA) or abscisic acid (ABA), although in the latter protein levels recover at the end of germination. While the levels of CDKA increase with IAA, they decrease with ABA. Both phytohormones, IAA and ABA, increase levels of CDKB1;1 only during the early germination times. CycD3;1 associated kinase activity is only reduced by both phytohormones towards the end of the germination period. On the other hand, lack of sucrose in the imbibition medium strongly reduces CycD3;1 protein levels without affecting the levels of neither CDKA nor CDKB1;1. The corresponding CycD3;1 associated kinase activity is also severely decreased. The presence of sucrose in the medium appears to stabilize the CycD3;1 protein levels. © 2016 Scandinavian Plant Physiology Society.

Flood replenishment: a new method of processor control.

PubMed

Frank, E D; Gray, J E; Wilken, D A

1980-01-01

In mechanized radiographic film processors that process medium to low volumes of film, roll films, and those that process single-emulsion films from nuclear medicine scans, computed tomography, and ultrasound, it is difficult to maintain the developer solution at a stable processing level. We describe our experience using flood replenishment, which is a method in which developer replenisher containing starter solution is introduced in the processor at timed intervals, independent of the number of films being processed. By this process, a stable level of developer activity is maintained in a processor used to develop a medium to low volume of single-emulsion film.

Freezing-Out Technique Applied to the Concentration of Biologically Active Materials

PubMed Central

Wilson, T. E.; Evans, D. J.; Theriot, Mary L.

1964-01-01

When applied to a dilute solution of folic acid and glucose, a freezing-out (with agitation) technique was shown to be an effective method of achieving a 20-fold reduction in volume with a loss of 10% of the active material being concentrated. Concentration of a stimulatory factor for Lactobacillus casei produced by Candida albicans in a complex medium was limited by the total solute concentration. Salts in the medium were concentrated to levels inhibitory for L. casei. The process is not selective and all solutes are concentrated. Images FIG. 2 FIG. 3 PMID:14131370

Role of manganese peroxidases and lignin peroxidases of Phanerochaete chrysosporium in the decolorization of kraft bleach plant effluent.

PubMed

Michel, F C; Dass, S B; Grulke, E A; Reddy, C A

1991-08-01

The role of lignin peroxidases (LIPs) and manganese peroxidases (MNPs) of Phanerochaete chrysosporium in decolorizing kraft bleach plant effluent (BPE) was investigated. Negligible BPE decolorization was exhibited by a per mutant, which lacks the ability to produce both the LIPs and the MNPs. Also, little decolorization was seen when the wild type was grown in high-nitrogen medium, in which the production of LIPs and MNPs is blocked. A lip mutant of P. chrysosporium, which produces MNPs but not LIPs, showed about 80% of the activity exhibited by the wild type, indicating that the MNPs play an important role in BPE decolorization. When P. chrysosporium was grown in a medium with 100 ppm of Mn(II), high levels of MNPs but no LIPs were produced, and this culture also exhibited high rates of BPE decolorization, lending further support to the idea that MNPs play a key role in BPE decolorization. When P. chrysosporium was grown in a medium with no Mn(II), high levels of LIPs but negligible levels of MNPs were produced and the rate and extent of BPE decolorization by such cultures were quite low, indicating that LIPs play a relatively minor role in BPE decolorization. Furthermore, high rates of BPE decolorization were seen on days 3 and 4 of incubation, when the cultures exhibit high levels of MNP activity but little or no LIP activity. These results indicate that MNPs play a relatively more important role than LIPs in BPE decolorization by P. chrysosporium.

Increased expression of tissue plasminogen activator and its inhibitor and reduced fibrinolytic potential of human endothelial cells cultured in elevated glucose.

PubMed

Maiello, M; Boeri, D; Podesta, F; Cagliero, E; Vichi, M; Odetti, P; Adezati, L; Lorenzi, M

1992-08-01

In diabetic patients, elevated plasma levels of t-PA and PAI-1 accompany impaired fibrinolysis. To identify mechanisms for these abnormalities, we examined whether vascular endothelial cells exposed to high glucose upregulate t-PA and PAI-1 production and whether ambient PA activity is decreased concomitantly. In 17 cultures of human umbilical vein endothelial cells grown to confluency in 30 mM glucose, the t-PA antigen released to the medium in 24 h was (median) 52 ng/10(6) cells (range 10-384) and the PAI-1 antigen was 872 ng/10(6) cells (range 217-2074)--both greater (P less than 0.02) than the amounts released by paired control cultures grown in 5 mM glucose--29 ng/10(6) cells (range 7.5-216) and 461 ng/10(6) cells (range 230-3215), respectively. In the presence of high glucose, the steady-state levels of t-PA and PAI-1 mRNAs were increased correspondingly (median 142 and 183% of control, respectively, P less than 0.05); high glucose per se and hypertonicity contributed to the upregulation in additive fashion. The PA activity of conditioned medium from cultures exposed to high glucose was 0.4 IU/ml (range 0.2-0.6), which was significantly lower (P less than 0.02) than the PA activity of control medium (0.5 IU/ml, range 0.2-0.9). No difference was observed when comparing the PA activities of acidified conditioned media, expected to be depleted of inhibitors. Thus, high glucose coordinately upregulates endothelial t-PA and PAI-1 expression through effects exerted at the pretranslational level and enhanced by even mild degrees of hypertonicity.(ABSTRACT TRUNCATED AT 250 WORDS)

Using Minute Papers to Determine Student Cognitive Development Levels

ERIC Educational Resources Information Center

Vella, Lia

2015-01-01

Can anonymous written feedback collected during classroom assessment activities be used to assess students' cognitive development levels? After library instruction in a first-year engineering design class, students submitted minute papers that included answers to "what they are left wondering." Responses were coded into low, medium and…

Drosophila melanogaster as a model system of aluminum toxicity and aging.

PubMed

Kijak, Ewelina; Rosato, Ezio; Knapczyk, Katarzyna; Pyza, Elżbieta

2014-04-01

The aim of this study was to investigate the toxic effects of aluminum (Al) on the model organism-Drosophila melanogaster. The study is especially concerned with the effects of aluminum on the fruit fly's development, life span, and circadian rhythm in rest and activity. Flies were exposed to aluminum in concentrations from 40 to 280 mg/kg in rearing media or the flies were raised on control medium. Moreover, the life span of insects exposed to aluminum containing 40, 120, or 240 mg/kg of Al in the medium, only during their larval development, during the whole life cycle and only in their adult life was tested. To check if aluminum and aging cause changes in D. melanogaster behavior, the locomotor activity of flies at different ages was recorded. Results showed that aluminum is toxic in concentrations above 160 mg/kg in the rearing medium. Depending on Al concentration and time of exposure, the life span of the flies was shortened. At intermediate concentrations (120 mg/kg), however, Al had a stimulating effect on males increasing their life span and level of locomotor activity. At higher concentration the aluminum exposure increased or decreased the level of locomotor activity of D. melanogaster depending on age of flies. In addition, in the oldest insects reared on aluminum supplemented media and in mid-aged flies reared on the highest concentration of Al the daily rhythm of activity was disrupted. © 2013 Institute of Zoology, Chinese Academy of Sciences.

Optimisation of the expression of a Trametes versicolor laccase gene in Pichia pastoris.

PubMed

O'Callaghan, J; O'Brien, M M; McClean, K; Dobson, A D W

2002-08-01

A cDNA encoding a laccase enzyme was isolated from a Trametes versicolor cDNA library. The gene was subcloned into the Pichia pastoris expression vector pPIC3.5 and transformed into the P. pastoris strains KM71 and GS115. Laccase-secreting transformants were selected by their ability to oxidise the substrate ABTS. No difference in laccase activity was observed between culture supernatants from GS115 (proteolytic) and KM71 (nonproteolytic) strains. The presence of at least 200 microM copper was necessary for optimal laccase activity in the culture supernatants. During growth of P. pastoris on minimal medium the pH of the medium was reduced to <3.0. If alanine was added to the medium the pH reduction was not as pronounced and at alanine concentrations >0.6% w/v the pH was kept constant for >7 days. Cultures in which the pH was maintained by alanine metabolism produced higher levels of laccase activity than those grown in the absence of alanine. This study describes the development of a medium that allows convenient pH control of P. pastoris without the need for continuous neutralisation.

Medium-Level Laser in Chronic Tinnitus Treatment

PubMed Central

Dejakum, K.; Piegger, J.; Plewka, C.; Gunkel, A.; Thumfart, W.; Kudaibergenova, S.; Goebel, G.; Kral, F.; Freysinger, W.

2013-01-01

The purpose of this study was to evaluate the effect of medium-level laser therapy in chronic tinnitus treatment. In a prospective double-blind placebo-controlled trial, either active laser (450 mW, 830 nm combined Ga-Al-As diode laser) or placebo irradiation was applied through the external acoustic meatus of the affected ear towards the cochlea. Fourty-eight patients with chronic tinnitus were studied. The main outcome was measured using the Goebel tinnitus questionnaire, visual analogue scales measuring the perceived loudness of tinnitus, the annoyance associated with tinnitus, and the degree of attention paid to tinnitus as well as psycho-acoustical matches of tinnitus pitch and loudness. The results did show only very moderate temporary improvement of tinnitus. Moreover, no statistically relevant differences between laser and placebo group could be found. We conclude that medium-level laser therapy cannot be regarded as an effective treatment of chronic tinnitus in our therapy regime considering the limited number of patients included in our study. PMID:24294604

Medium-level laser in chronic tinnitus treatment.

PubMed

Dejakum, K; Piegger, J; Plewka, C; Gunkel, A; Thumfart, W; Kudaibergenova, S; Goebel, G; Kral, F; Freysinger, W

2013-01-01

The purpose of this study was to evaluate the effect of medium-level laser therapy in chronic tinnitus treatment. In a prospective double-blind placebo-controlled trial, either active laser (450 mW, 830 nm combined Ga-Al-As diode laser) or placebo irradiation was applied through the external acoustic meatus of the affected ear towards the cochlea. Fourty-eight patients with chronic tinnitus were studied. The main outcome was measured using the Goebel tinnitus questionnaire, visual analogue scales measuring the perceived loudness of tinnitus, the annoyance associated with tinnitus, and the degree of attention paid to tinnitus as well as psycho-acoustical matches of tinnitus pitch and loudness. The results did show only very moderate temporary improvement of tinnitus. Moreover, no statistically relevant differences between laser and placebo group could be found. We conclude that medium-level laser therapy cannot be regarded as an effective treatment of chronic tinnitus in our therapy regime considering the limited number of patients included in our study.

Muscle activity during leg strengthening exercise using free weights and elastic resistance: effects of ballistic vs controlled contractions.

PubMed

Jakobsen, Markus Due; Sundstrup, Emil; Andersen, Christoffer H; Aagaard, Per; Andersen, Lars L

2013-02-01

The present study's aim was to evaluate muscle activity during leg exercises using elastic vs. isoinertial resistance at different exertion and loading levels, respectively. Twenty-four women and eighteen men aged 26-67 years volunteered to participate in the experiment. Electromyographic (EMG) activity was recorded in nine muscles during a standardized forward lunge movement performed with dumbbells and elastic bands during (1) ballistic vs. controlled exertion, and (2) at low, medium and high loads (33%, 66% and 100% of 10 RM, respectively). The recorded EMG signals were normalized to MVC EMG. Knee joint angle was measured using electronic inclinometers. The following results were obtained. Loading intensity affected EMG amplitude in the order: low

Disruption of plastid acyl:acyl carrier protein synthetases increases medium chain fatty acid accumulation in seeds of transgenic Arabidopsis.

PubMed

Tjellström, Henrik; Strawsine, Merissa; Silva, Jillian; Cahoon, Edgar B; Ohlrogge, John B

2013-04-02

Engineering transgenic plants that accumulate high levels of medium-chain fatty acids (MCFA) has been least successful for shorter chain lengths (e.g., C8). We demonstrate that one limitation is the activity of acyl-ACP synthetase (AAE) that re-activates fatty acids released by acyl-ACP thioesterases. Seed expression of Cuphea pulcherrima FATB acyl-ACP thioesterase in a double mutant lacking AAE15/16 increased 8:0 accumulation almost 2-fold compared to expression in wild type. These results also provide an in planta demonstration that AAE enzymes participate not only in activation of exogenously added MCFA but also in activation of MCFA synthesized in plastids. Copyright © 2013 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

Hair-growth stimulation by conditioned medium from vitamin D3-activated preadipocytes in C57BL/6 mice.

PubMed

Jung, Min Kyung; Ha, Soogyeong; Huh, Scarlett Yoona; Park, Seung Beom; Kim, Sangyoon; Yang, Yoolhee; Kim, Daejin; Hur, Dae Young; Jeong, Hyuk; Bang, Sa Ik; Park, Hyunjeong; Cho, Daeho

2015-05-01

Recently, immature adipocyte lineage cells have been suggested as a potential hair-growth stimulator. Diverse studies have been attempted to find methods for the preconditioning of immature adipocyte lineage cells. The present study investigates the effect of conditioned medium (CM) from vitamin D3 (Vd3) pre-activated preadipocytes on hair-growth ability. To test the effect of CM from Vd3 pre-activated preadipocytes on hair-growth efficiency in mice, we compared the differences in hair regenerated after injecting CM from mouse preadipocytes pre-activated with or without Vd3. Next, to determine the regulating factors, the VEGF level was measured by ELISA and angiogenesis level was evaluated by IHC. Finally, the signaling mechanism was investigated by inhibitor kinase assay and western blotting. The CM from Vd3 pre-activated preadipocyte injection markedly promoted the ability of hair regeneration in mice. The VEGF levels were increased by Vd3 treatment in vitro and the CM from Vd3 pre-activated preadipocytes significantly increased the angiogenesis in vivo, suggesting the involvement of angiognensis in the hair regeneration induced by CM from pre-activated preadipocytes. In signaling study, Vd3-enhanced VEGF production was reduced by an ERK1/2 inhibitor and the level of ERK1/2 phosphorylation was increased by treatment with Vd3. This has been the first report on CM from Vd3 pre-activated preadipocyte displaying stimulatory effects on hair growth via the enhancement of angiogenesis in a hairless-induced C57BL/6 mice. Copyright © 2015 Elsevier Inc. All rights reserved.

The neural correlates of cognitive effort in anxiety: effects on processing efficiency.

PubMed

Ansari, Tahereh L; Derakshan, Nazanin

2011-03-01

We investigated the neural correlates of cognitive effort/pre-target preparation (Contingent Negative Variation activity; CNV) in anxiety using a mixed antisaccade task that manipulated the interval between offset of instructional cue and onset of target (CTI). According to attentional control theory (Eysenck et al., 2007) we predicted that anxiety should result in increased levels of compensatory effort, as indicated by greater frontal CNV, to maintain comparable levels of performance under competing task demands. Our results showed that anxiety resulted in faster antisaccade latencies during medium compared with short and long CTIs. Accordingly, high-anxious individuals compared with low-anxious individuals showed greater levels of CNV activity at frontal sites during medium CTI suggesting that they exerted greater cognitive effort and invested more attentional resources in preparation for the task goal. Our results are the first to demonstrate the neural correlates of processing efficiency and compensatory effort in anxiety and are discussed within the framework of attentional control theory. Copyright © 2011 Elsevier B.V. All rights reserved.

NMDA receptor gating of information flow through the striatum in vivo.

PubMed

Pomata, Pablo E; Belluscio, Mariano A; Riquelme, Luis A; Murer, M Gustavo

2008-12-10

A role of NMDA receptors in corticostriatal synaptic plasticity is widely acknowledged. However, the conditions that allow NMDA receptor activation in the striatum in vivo remain obscure. Here we show that NMDA receptors contribute to sustain the membrane potential of striatal medium spiny projection neurons close to threshold during spontaneous UP states in vivo. Moreover, we found that the blockade of striatal NMDA receptors reduces markedly the spontaneous firing of ensembles of medium spiny neurons during slow waves in urethane-anesthetized rats. We speculate that recurrent activation of NMDA receptors during UP states allows off-line information flow through the striatum and system level consolidation during habit formation.

Zinc is released by cultured astrocytes as a gliotransmitter under hypoosmotic stress-loaded conditions and regulates microglial activity.

PubMed

Segawa, Shohei; Nishiura, Takeshi; Furuta, Takahiro; Ohsato, Yuki; Tani, Misaki; Nishida, Kentaro; Nagasawa, Kazuki

2014-01-17

Astrocytes contribute to the maintenance of brain homeostasis via the release of gliotransmitters such as ATP and glutamate. Here we examined whether zinc was released from astrocytes under stress-loaded conditions, and was involved in the regulation of microglial activity as a gliotransmitter. Hypoosmotic stress was loaded to astrocytes using balanced salt solution prepared to 214-314 mOsmol/L, and then intra- and extra-cellular zinc levels were assessed using Newport Green DCF diacetate (NG) and ICP-MS, respectively. Microglial activation by the astrocytic supernatant was assessed by their morphological changes and poly(ADP-ribose) (PAR) polymer accumulation. Exposure of astrocytes to hypoosmotic buffer, increased the extracellular ATP level in osmolarity-dependent manners, indicating a load of hypoosmotic stress. In hypoosmotic stress-loaded astrocytes, there were apparent increases in the intra- and extra-cellular zinc levels. Incubation of microglia in the astrocytic conditioned medium transformed them into the activated "amoeboid" form and induced PAR formation. Administration of an extracellular zinc chelator, CaEDTA, to the astrocytic conditioned medium almost completely prevented the microglial activation. Treatment of astrocytes with an intracellular zinc chelator, TPEN, suppressed the hypoosmotic stress-increased intracellular, but not the extracellular, zinc level, and the increase in the intracellular zinc level was blocked partially by a nitric oxide synthase inhibitor, but not by CaEDTA, indicating that the mechanisms underlying the increases in the intra- and extra-cellular zinc levels might be different. These findings suggest that under hypoosmotic stress-loaded conditions, zinc is released from astrocytes and then plays a primary role in microglial activation as a gliotransmitter. Copyright © 2013 Elsevier Inc. All rights reserved.

Character Education for Cyberspace: Developing Good Netizens

ERIC Educational Resources Information Center

Milson, Andrew J.; Chu, Beong-Wan

2002-01-01

At the simplest level, cyberspace is a sea of information that functions as a huge, generally disorganized, database. At a more complex level, cyberspace functions as a communication medium. In the most complex sense, cyberspace may be conceived as a parallel world where people can conduct ordinary activities without engaging in the physical acts…

The Effects of Bench Press Variations in Competitive Athletes on Muscle Activity and Performance

PubMed Central

Saeterbakken, Atle Hole; Mo, Dag-André; Scott, Suzanne; Andersen, Vidar

2017-01-01

Abstract The aim of the study was to compare the EMG activity performing 6RM competition style bench press (flat bench-wide grip) with 1) medium and narrow grip widths on a flat bench and 1) inclined and declined bench positions with a wide grip. Twelve bench press athletes competing at national and international level participated in the study. EMG activity was measured in the pectoralis major, anterior and posterior deltoid, biceps brachii, triceps brachii and latissimus dorsi. Non-significant differences in activation were observed between the three bench positions with the exception of 58.5-62.6% lower triceps brachii activation, but 48.3-68.7% greater biceps brachii activation in the inclined bench compared with the flat and declined bench position. Comparing the three grip widths, non-significant differences in activations were observed, with the exception of 25.9-30.5% lower EMG activity in the biceps brachii using a narrow grip, compared to the medium and wide grip conditions. The 6-RM loads were 5.8-11.1% greater using a medium and wide grip compared to narrow grip width and 18.5-21.5% lower in the inclined bench position compared with flat and declined. Comparing the EMG activity during the competition bench press style with either the inclined and declined bench position (wide grip) or using a narrow and medium grip (flat bench), only resulted in different EMG activity in the biceps- and triceps brachii. The 6RM loads varied with each bench press variation and we recommend the use of a wide grip on a flat bench during high load hypertrophy training to bench press athletes. PMID:28713459

The Effects of Bench Press Variations in Competitive Athletes on Muscle Activity and Performance.

PubMed

Saeterbakken, Atle Hole; Mo, Dag-André; Scott, Suzanne; Andersen, Vidar

2017-06-01

The aim of the study was to compare the EMG activity performing 6RM competition style bench press (flat bench-wide grip) with 1) medium and narrow grip widths on a flat bench and 1) inclined and declined bench positions with a wide grip. Twelve bench press athletes competing at national and international level participated in the study. EMG activity was measured in the pectoralis major, anterior and posterior deltoid, biceps brachii, triceps brachii and latissimus dorsi. Non-significant differences in activation were observed between the three bench positions with the exception of 58.5-62.6% lower triceps brachii activation, but 48.3-68.7% greater biceps brachii activation in the inclined bench compared with the flat and declined bench position. Comparing the three grip widths, non-significant differences in activations were observed, with the exception of 25.9-30.5% lower EMG activity in the biceps brachii using a narrow grip, compared to the medium and wide grip conditions. The 6-RM loads were 5.8-11.1% greater using a medium and wide grip compared to narrow grip width and 18.5-21.5% lower in the inclined bench position compared with flat and declined. Comparing the EMG activity during the competition bench press style with either the inclined and declined bench position (wide grip) or using a narrow and medium grip (flat bench), only resulted in different EMG activity in the biceps- and triceps brachii. The 6RM loads varied with each bench press variation and we recommend the use of a wide grip on a flat bench during high load hypertrophy training to bench press athletes.

Antitumor Activity of Rat Mesenchymal Stem Cells during Direct or Indirect Co-Culturing with C6 Glioma Cells.

PubMed

Gabashvili, A N; Baklaushev, V P; Grinenko, N F; Mel'nikov, P A; Cherepanov, S A; Levinsky, A B; Chehonin, V P

2016-02-01

The tumor-suppressive effect of rat mesenchymal stem cells against low-differentiated rat C6 glioma cells during their direct and indirect co-culturing and during culturing of C6 glioma cells in the medium conditioned by mesenchymal stem cells was studied in an in vitro experiment. The most pronounced antitumor activity of mesenchymal stem cells was observed during direct co-culturing with C6 glioma cells. The number of live C6 glioma cells during indirect co-culturing and during culturing in conditioned medium was slightly higher than during direct co-culturing, but significantly differed from the control (C6 glioma cells cultured in medium conditioned by C6 glioma cells). The cytotoxic effect of medium conditioned by mesenchymal stem cells was not related to medium depletion by glioma cells during their growth. The medium conditioned by other "non-stem" cells (rat astrocytes and fibroblasts) produced no tumor-suppressive effect. Rat mesenchymal stem cells, similar to rat C6 glioma cells express connexin 43, the main astroglial gap junction protein. During co-culturing, mesenchymal stem cells and glioma C6 cells formed functionally active gap junctions. Gap junction blockade with connexon inhibitor carbenoxolone attenuated the antitumor effect observed during direct co-culturing of C6 glioma cells and mesenchymal stem cells to the level produced by conditioned medium. Cell-cell signaling mediated by gap junctions can be a mechanism of the tumor-suppressive effect of mesenchymal stem cells against C6 glioma cells. This phenomenon can be used for the development of new methods of cell therapy for high-grade malignant gliomas.

Antioxidant capacity of 3D human skin EpiDerm model: effects of skin moisturizers.

PubMed

Grazul-Bilska, A T; Bilski, J J; Redmer, D A; Reynolds, L P; Abdullah, K M; Abdullah, A

2009-06-01

The objective of this study was to determine the effects of skin moisturizers on total antioxidant capacity (TAC) of human skin using EpiDerm model. Three different skin moisturizers containing antioxidant ingredients (samples 1-3) or aloe vera extract were topically applied to EpiDerm units and incubated for 2 and 24 h to determine acute and longer-term effects of applied samples on TAC and glutathione peroxidase activity in medium and/or homogenized skin tissues. Total antioxidant capacity in medium and skin homogenates was enhanced (P < 0.0001) by gel containing antioxidant ingredients (sample 2) after 2 and 24 h of incubation. Total antioxidant capacity in medium was also enhanced (P < 0.001) by cream containing antioxidant ingredients (sample 3) after 24 h of incubation. Overall, TAC in medium was greater (P < 0.02) after 24 h than 2 h of incubation. Skin moisturizer cream with high antioxidant levels determined by using oxygen radical absorbance capacity testing (sample 1) and aloe vera extract did not affect TAC. Glutathione peroxidase activity was enhanced (P < 0.0001) in medium and skin homogenates by sample 2 but not by any other sample. These data demonstrate high potential of gel and cream (samples 2 and 3) containing antioxidant ingredients in enhancing antioxidant capacity of EpiDerm which will likely contribute to overall skin health. Results of this experiment will help to better understand mechanisms of effects of skin moisturizers containing antioxidant ingredients on skin function at the tissue level and to establish effective strategies for skin protection and clinical treatments of skin disorders and possibly healing wounds.

Physical structure changes of solid medium by steam explosion sterilization.

PubMed

Zhao, Zhi-Min; Wang, Lan; Chen, Hong-Zhang

2016-03-01

Physical structure changes of solid medium were investigated to reveal effects of steam explosion sterilization on solid-state fermentation (SSF). Results indicated that steam explosion changed the structure of solid medium at both molecular and three-dimensional structural levels, which exposed hydrophilic groups and enlarged pores and cavities. It was interesting to find that pores where capillary water located were the active sites for SSF, due to the close relationship among capillary water relaxation time, specific surface area and fermentation performance. Therefore, steam explosion sterilization increased the effective contact area for microbial cells on solid medium, which contributed to improving SSF performance. Combined with the previous research, mechanisms of SSF improvement by steam explosion sterilization contained both chemical and physical effects. Copyright © 2015 Elsevier Ltd. All rights reserved.

Adaptive Management Approach to Oil and Gas Activities in Areas Occupied by Pacific Walrus

NASA Astrophysics Data System (ADS)

Ireland, D.; Broker, K.; San Filippo, V.; Brzuzy, L.; Morse, L.

2016-12-01

During Shell's 2015 exploration drilling program in the Chukchi Sea, activities were conducted in accordance with a Letter of Authorization issued by the United States Fish and Wildlife Service that allowed the incidental harassment of Pacific Walrus and Polar Bears under the Marine Mammal Protection Act. As a part of the request for authorization, Shell proposed a process to monitor and assess the potential for activities to interact with walruses on ice, especially if ice posed a potential threat to the drill site. The process assimilated near real-time information from multiple data sources including vessel-based observations, aerial surveys, satellite-linked GPS tags on walrus, and satellite imagery of ice conditions and movements. These data were reviewed daily and assessed in the context of planned activities to assign a risk level (low, medium, or high). The risk level was communicated to all assets in the field and decision makers during morning briefings. A low risk level meant that planned activities could occur without further review. A medium risk level meant that some operations had a greater potential of interacting with walrus on ice and that additional discussions of those activities were required to determine the relative risk of potential impacts compare to the importance of the planned activity. A high risk level meant that the planned activities were necessary and walrus on ice were likely to be encountered. Assignment of a high risk level triggered contact with agency personnel and directly incorporated them into the assessment and decision making process. This process made effective use of relevant available information to provide meaningful assessments at temporal and spatial scales that allowed approved activities to proceed while minimizing potential impacts. More so, this process provides a valuable alternative to large-scale restriction areas with coarse temporal resolution without reducing protection to target species.

Adaptive Management Approach to Oil and Gas Activities in Areas Occupied by Pacific Walrus

NASA Astrophysics Data System (ADS)

Ireland, D.; Broker, K.; San Filippo, V.; Brzuzy, L.; Morse, L.

2016-02-01

During Shell's 2015 exploration drilling program in the Chukchi Sea, activities were conducted in accordance with a Letter of Authorization issued by the United States Fish and Wildlife Service that allowed the incidental harassment of Pacific Walrus and Polar Bears under the Marine Mammal Protection Act. As a part of the request for authorization, Shell proposed a process to monitor and assess the potential for activities to interact with walruses on ice, especially if ice posed a potential threat to the drill site. The process assimilated near real-time information from multiple data sources including vessel-based observations, aerial surveys, satellite-linked GPS tags on walrus, and satellite imagery of ice conditions and movements. These data were reviewed daily and assessed in the context of planned activities to assign a risk level (low, medium, or high). The risk level was communicated to all assets in the field and decision makers during morning briefings. A low risk level meant that planned activities could occur without further review. A medium risk level meant that some operations had a greater potential of interacting with walrus on ice and that additional discussions of those activities were required to determine the relative risk of potential impacts compare to the importance of the planned activity. A high risk level meant that the planned activities were necessary and walrus on ice were likely to be encountered. Assignment of a high risk level triggered contact with agency personnel and directly incorporated them into the assessment and decision making process. This process made effective use of relevant available information to provide meaningful assessments at temporal and spatial scales that allowed approved activities to proceed while minimizing potential impacts. More so, this process provides a valuable alternative to large-scale restriction areas with coarse temporal resolution without reducing protection to target species.

Riluzole in the prelimbic medial prefrontal cortex attenuates veratrine-induced anxiety-like behaviors in mice.

PubMed

Ohashi, Masanori; Saitoh, Akiyoshi; Yamada, Misa; Oka, Jun-Ichiro; Yamada, Mitsuhiko

2015-01-01

We previously demonstrated in mice that the activation of prelimbic medial prefrontal cortex (PL) with the sodium channel activator veratrine induces anxiety-like behaviors via NMDA receptor-mediated glutamatergic neurotransmission. Riluzole directly affects the glutamatergic system and has recently been suggested to have an anxiolytic-like effect in both experimental animals and patients with anxiety disorders. We investigated the effects of co-perfusion of riluzole on veratrine-induced anxiety-like behaviors in mice. Extracellular glutamate levels were measured in 7-week-old male C57BL6 mice by using an in vivo microdialysis-HPLC/ECD system, and behaviors were assessed simultaneously in an open field (OF) test. Basal levels of glutamate were measured by collecting samples every 10 min for 60 min. The medium containing drugs was perfused for 30 min, and the OF test was performed during the last 10 min of drug perfusion. After the drug treatments, the drug-containing medium was switched to perfusion of control medium lacking drugs, and then samples were collected for another 90 min. Riluzole co-perfusion attenuated veratrine-induced increase in extracellular glutamate levels in the PL and completely diminished veratrine-induced anxiety-like behaviors. Interestingly, riluzole perfusion alone in the PL did not affect the basal levels of glutamate and anxiety-like behaviors. Our results suggest that compounds like riluzole that inhibit glutamatergic function in the PL are possible candidates for novel anxiolytics.

Influence of culture medium growth variables on Ganoderma lucidum exopolysaccharides structural features.

PubMed

Fraga, Irene; Coutinho, João; Bezerra, Rui M; Dias, Albino A; Marques, Guilhermina; Nunes, Fernando M

2014-10-13

In this work the effect of carbon and nitrogen levels and initial pH of the wheat extract culture medium of submerged culture of Ganoderma lucidum on the amount, purity and structural features of exopolysaccharides (EPS) were studied. A low peptone level (1.65 g L(-1)) favored mycelium biomass, EPS purity, but a higher supply of peptone (4.80 g L(-1)) is needed for maximum EPS production. The carbohydrate composition of the EPS and structural features also changed significantly according to the different growing conditions, being observed significant differences in the (1 → 3)/(1 → 4)-Glcp ratio and also on the branching degree of EPS. As the biological activities of EPS are highly dependent on the polysaccharide structural features, this variability can have implications on the EPS biological activities, but can also be used advantageously to produce tailor made polysaccharides with specific applications. Copyright © 2014 Elsevier Ltd. All rights reserved.

Development of markers for cholinergic neurones in re-aggregate cultures of foetal rat whole brain in serum-containing and serum-free media: effects of triiodothyronine (T3).

PubMed Central

Atterwill, C. K.; Kingsbury, A.; Nicholls, J.; Prince, A.

1984-01-01

Development has been studied in re-aggregate cultures derived from the 16 day foetal rat brain and the effects of triiodothyronine (T3) investigated. Cultures were maintained in either a medium containing 10% serum (S+), or in serum-free culture medium (S-) or in serum-free medium containing 30nM T3. The muscarinic cholinoceptor, measured by specific binding of [3H]-quinuclidinyl benzitate ([3H]-QNB) at 9 and 14 days in vitro, was at a lower level in the serum-free cultured cells compared with those in serum-containing culture medium (S+). In cultures in the latter medium, receptor concentration at day 14 was of a similar magnitude to that in rat brain at an equivalent postnatal age. Binding increased with development from 9 to 14 days in vitro in the S+ medium but not in the S- medium. T3 treatment caused an 85% increase in [3H]-QNB binding compared with the cultures in S- medium at day 14 to a level equivalent to that found in the cells grown in S+ medium. This increase was reflected in the Bmax but not in the KD (approx. 0.1nM). Choline acetyltransferase (ChAT) activity developed more slowly in the S- medium than in the S+ medium where the specific activity approximated values obtained in vivo. T3 treatment of cultures grow in S- medium significantly enhanced the developmental rate of increase of ChAT activity. The characteristics of [3H]-choline uptake and metabolism in the cultures was examined. Uptake was strictly Na+-independent but was energy-dependent, and inhibited by 2, 4'-dinitrophenol (2, 4'-DNP) and cooling (0-4 degrees C). Neither iodoacetate nor ouabain had any effect on the amount of uptake. Hemicholinium (HC3) was a potent inhibitor of uptake (70% inhibition at 10 microM HC3). Metabolism studies showed virtually no conversion to [3H]-acetylcholine ([3H]-ACH) in reaggregates grown in either the S+, S- or T3 containing media. However, a small amount of [3H]-choline was incorporated into phosphorylcholine. T3 treatment had no effect on this metabolic profile. The kinetics of [3H]-choline uptake by the re-aggregates was also studied in the re-aggregate cultures (after 12 and 22 days in vitro) using [3H]-choline at 0.05-100 microM. Both Eadie-Hofstee transformation and least-squares analysis of the data showed that the uptake comprised only a single low-affinity component with an apparent Kt = approx. 50 microM. Unlike ChAT and [3H]-QNB binding, there appeared to be no difference between the uptake in the different culture conditions.(ABSTRACT TRUNCATED AT 400 WORDS) Images Fig. 6 PMID:6487898

Decolorization of RBBR by plant cells and correlation with the transformation of PCBs.

PubMed

Chroma, Ludmila; Macek, Tomas; Demnerova, Katerina; Macková, Martina

2002-11-01

An extracellular H2O2-requiring Remazol Brilliant Blue R (RBBR) decolorizing enzyme activity was detected after cultivation of cells of various plant species both in liquid medium and when growing on agar plates containing RBBR. Level of the enzyme activity was compared with the ability to metabolize polychlorinated biphenyls (PCBs). The ability to decolorize RBBR was tested in the presence and absence of PCBs. The cultures with high PCB-transforming activity proved to exhibit RBBR oxidase much more resistant towards the influence of PCBs. In addition low activities of lignin peroxidase (LiP) and manganese dependent peroxidase (MnP) were detected in medium and in plant cells. No correlation of MnP and LiP activities with PCB degradation could be found. The RBBR decolorization could be used as a rough screening method for plant cultures able to metabolize PCBs.

MicroRNAs, polyamines, and the activities antioxidant enzymes are associated with in vitro rooting in white pine (Pinus strobus L.).

PubMed

Fei, Yunjun; Xiao, Bo; Yang, Man; Ding, Qiong; Tang, Wei

2016-01-01

Molecular mechanism of in vitro rooting in conifer is not fully understood. After establishment of a regeneration procedure in eastern white pine (Pinus strobus L.) using mature embryos as explants to induce shoot formation on medium containing 3 μM IAA, 6 μM BA and 6 μM TDZ and induce root formation on medium containing 0.001-0.05 μM IAA, 0.001-0.05 μM IBA, 0.001-0.05 μM TDZ, we have investigated the changes of polyamine content and the activities of antioxidant enzymes during in vitro rooting in P. strobus. Our results demonstrated that putrescine (Put), spermidine (Spd), and spermine (Spm) did not increase in P. strobus during the first week of rooting on medium supplemented with 0.01 μM indole-3-acetic acid (IAA), whereas the levels of Put, Spd, and Spm increased during the 1st-3rd week of culture on medium with IAA, and then decreased on medium with IAA. No such a change in Put, Spd, and Spm was observed on medium without IAA. Measurement of antioxidant enzyme activity demonstrated that the activities of polyphenol oxidase, catalase, and peroxidase slightly increased in the first week of culture and reached to the highest peak in the 3rd-5th week of culture. Quantitative RT-PCR results indicated that miR160 was increased on the 7th day, miR162, miR397, and miR408 was increased from the 21th to 35th day, miR857 was increased on the 35th day, and miR827 was increased on the 49th day. These results demonstrated that enhanced polyamine biosynthesis, antioxidant enzyme activity, and microRNAs are correlated with the root induction and formation in P. strobus.

A case study of the response of the magnetosphere to changes in the interplanetary medium

NASA Technical Reports Server (NTRS)

Rostoker, G.; Baumjohann, W.; Russell, C. T.

1983-01-01

A detailed analysis of world-wide ground based magnetometer data is presented, together with information on the plasma and magnetic field properties of the interplanetary medium and magnetosheath obtained from the ISEE 1 and 2 and IMP 8 spacecraft. The event concerned exhibited an interval of relatively stable southward IMF followed by a sharp northward turning. It is pointed out that during the interval of southward IMF there were occasional transient northward turnings with significant substorm expansive phase activity appearing to be triggered by these transient northward turnings. The final northward turning of the IMF was linked with an episode of strong magnetospheric substorm expansive phase activity after which the level of high latitude magnetic activity declined to a low level. Evidence is presented indicating that the driven system auroral electrojets begin to decay at the time of the northward turning of the IMF, even as the substorm expansive phase activity is initiated in the midnight sector. The collapse of the substorm current wedge during the final decay of high latitude activity is described in some detail, and it is shown that this collapse occurs progressively from east to west in a series of impulsive episodes.

Kidney transplantation: a systematic review of interventional and observational studies of physical activity on intermediate outcomes.

PubMed

Macdonald, Jamie Hugo; Kirkman, Danielle; Jibani, Mahdi

2009-11-01

Kidney transplant patients have decreased quality and longevity of life. Whether exercise can positively affect associated outcomes such as physical functioning, metabolic syndrome, kidney function, and immune function, has only been addressed in relatively small studies. Thus the aim of this systematic review was to determine effects of physical activity level on these intermediate outcomes in kidney transplant patients. We electronically and hand searched to identify 21 studies (6 retrospective assessments of habitual physical activity and 15 intervention studies including 6 controlled trials). After study quality assessment, intermediate outcomes associated with quality and longevity of life were expressed as correlations or percentage changes in addition to effect sizes. Habitual physical activity level was positively associated with quality of life and aerobic fitness and negatively associated with body fat (medium to large effect sizes). Exercise interventions also showed medium to large positive effects on aerobic capacity (10%-114% increase) and muscle strength (10%-22% increase). However, exercise programs had minimal or contradictory effects on metabolic syndrome and immune and kidney function. In kidney transplant patients, physical activity intervention is warranted to enhance physical functioning. Whether exercise impacts on outcomes associated with longevity of life requires further study.

Glutamine Acts as a Neuroprotectant against DNA Damage, Beta-Amyloid and H2O2-Induced Stress

PubMed Central

Chen, Jianmin; Herrup, Karl

2012-01-01

Glutamine is the most abundant free amino acid in the human blood stream and is ‘conditionally essential’ to cells. Its intracellular levels are regulated both by the uptake of extracellular glutamine via specific transport systems and by its intracellular synthesis by glutamine synthetase (GS). Adding to the regulatory complexity, when extracellular glutamine is reduced GS protein levels rise. Unfortunately, this excess GS can be maladaptive. GS overexpression is neurotoxic especially if the cells are in a low-glutamine medium. Similarly, in low glutamine, the levels of multiple stress response proteins are reduced rendering cells hypersensitive to H2O2, zinc salts and DNA damage. These altered responses may have particular relevance to neurodegenerative diseases of aging. GS activity and glutamine levels are lower in the Alzheimer's disease (AD) brain, and a fraction of AD hippocampal neurons have dramatically increased GS levels compared with control subjects. We validated the importance of these observations by showing that raising glutamine levels in the medium protects cultured neuronal cells against the amyloid peptide, Aβ. Further, a 10-day course of dietary glutamine supplementation reduced inflammation-induced neuronal cell cycle activation, tau phosphorylation and ATM-activation in two different mouse models of familial AD while raising the levels of two synaptic proteins, VAMP2 and synaptophysin. Together, our observations suggest that healthy neuronal cells require both intracellular and extracellular glutamine, and that the neuroprotective effects of glutamine supplementation may prove beneficial in the treatment of AD. PMID:22413000

Chitin synthesis in Saccharomyces cerevisiae in response to supplementation of growth medium with glucosamine and cell wall stress.

PubMed

Bulik, Dorota A; Olczak, Mariusz; Lucero, Hector A; Osmond, Barbara C; Robbins, Phillips W; Specht, Charles A

2003-10-01

In Saccharomyces cerevisiae most chitin is synthesized by Chs3p, which deposits chitin in the lateral cell wall and in the bud-neck region during cell division. We have recently found that addition of glucosamine (GlcN) to the growth medium leads to a three- to fourfold increase in cell wall chitin levels. We compared this result to the increases in cellular chitin levels associated with cell wall stress and with treatment of yeast with mating pheromone. Since all three phenomena lead to increases in precursors of chitin, we hypothesized that chitin synthesis is at least in part directly regulated by the size of this pool. This hypothesis was strengthened by our finding that addition of GlcN to the growth medium causes a rapid increase in chitin synthesis without any pronounced change in the expression of more than 6,000 genes monitored with Affymetrix gene expression chips. In other studies we found that the specific activity of Chs3p is higher in the total membrane fractions from cells grown in GlcN and from mutants with weakened cell walls. Sucrose gradient analysis shows that Chs3p is present in an inactive form in what may be Golgi compartments but as an active enzyme in other intracellular membrane-bound vesicles, as well as in the plasma membrane. We conclude that Chs3p-dependent chitin synthesis in S. cerevisiae is regulated both by the levels of intermediates of the UDP-GlcNAc biosynthetic pathway and by an increase in the activity of the enzyme in the plasma membrane.

The behavior of seasonal variations in induced seismicity in the Koyna-Warna region, western India

NASA Astrophysics Data System (ADS)

Smirnov, V. B.; Srinagesh, D.; Ponomarev, A. V.; Chadha, R.; Mikhailov, V. O.; Potanina, M. G.; Kartashov, I. M.; Stroganova, S. M.

2017-07-01

Based on the earthquake catalog data for the Koyna-Warna region of induced seismicity in western India, the seasonal variations in seismic activity associated with annual fluctuations in the reservoir water level are analyzed over the time span of the entire history of seismological observations in this region. The regularities in the time changes in the structure of seasonal variations are revealed. The seasonal seismic activity is minimal in May-June when the reservoir level is lowest. During the remaining part of the year, the activity has three peaks: the fall peak in September, winter peak in November-December, and spring peak in February-March. The first mentioned peak, which falls in the phase of the water level reaching its maximal seasonal value is considered as the immediate response of the fluid saturated medium to the additional loading under the weight of reservoir water. The two subsequent maxima concur with the decline phase in the reservoir level and are interpreted as the delayed response associated with the changes in the properties of the medium due to water diffusion. It is shown that the intensities of the immediate and delayed responses to the seasonal water level variations both vary with time as does their ratio. The probable factors affecting the variations in the intensity of the seasonal components of the reservoir-induced seismicity are discussed.

Alterations in protein kinase C activity and processing during zinc-deficiency-induced cell death.

PubMed

Chou, Susan S; Clegg, Michael S; Momma, Tony Y; Niles, Brad J; Duffy, Jodie Y; Daston, George P; Keen, Carl L

2004-10-01

Protein kinases C (PKCs) are a family of serine/threonine kinases that are critical for signal transduction pathways involved in growth, differentiation and cell death. All PKC isoforms have four conserved domains, C1-C4. The C1 domain contains cysteine-rich finger-like motifs, which bind two zinc atoms. The zinc-finger motifs modulate diacylglycerol binding; thus, intracellular zinc concentrations could influence the activity and localization of PKC family members. 3T3 cells were cultured in zinc-deficient or zinc-supplemented medium for up to 32 h. Cells cultured in zinc-deficient medium had decreased zinc content, lowered cytosolic classical PKC activity, increased caspase-3 processing and activity, and reduced cell number. Zinc-deficient cytosols had decreased activity and expression levels of PKC-alpha, whereas PKC-alpha phosphorylation was not altered. Inhibition of PKC-alpha with Gö6976 had no effect on cell number in the zinc-deficient group. Proteolysis of the novel PKC family member, PKC-delta, to its 40-kDa catalytic fragment occurred in cells cultured in the zinc-deficient medium. Occurrence of the PKC-delta fragment in mitochondria was co-incident with caspase-3 activation. Addition of the PKC-delta inhibitor, rottlerin, or zinc to deficient medium reduced or eliminated proteolysis of PKC-delta, activated caspase-3 and restored cell number. Inhibition of caspase-3 processing by Z-DQMD-FMK (Z-Asp-Gln-Met-Asp-fluoromethylketone) did not restore cell number in the zinc-deficient group, but resulted in processing of full-length PKC-delta to a 56-kDa fragment. These results support the concept that intracellular zinc concentrations influence PKC activity and processing, and that zinc-deficiency-induced apoptosis occurs in part through PKC-dependent pathways.

Influence of environmental temperature on in vivo energy expenditure in vitro ouabain-sensitive respiration in duodenal mucosa and liver in rats fed different levels of dietary fiber or protein.

PubMed

Jørgensen, H; Zhao, X Q

1997-12-01

Seventy two Wistar rats were used in two repeat studies to investigate the effect of environmental temperature (18 degrees C or 28 degrees C) and increasing levels of dietary fibre (low, 68 g/kg DM; medium 110 g/kg DM; high, 157 g/kg DM) or protein (low, 91 g/kg DM; medium, 171 g/kg DM; high, 262 g/kg DM) on digestive tract, visceral organ size, energy metabolism, and respiration attributable to Na+,K(+)-ATPase activity in duodenal mucosa and liver. Total and ouabain-sensitive (a measure of Na+,K(+)-ATPase activity) O2 consumption in vitro of tissues were measured polarographically using a Clark-style YSI biological O2 monitor. Whole body heat production (in vivo) was measured using open-circuit respiration chambers. The weight of the visceral organs was higher in rats housed at 18 degrees C than at 28 degrees C. The empty weight of the small intestine, caecum, and colon increased as the level of dietary fibre increased (P 0.05). Heat production as a proportion of metabolizable energy was higher (P < 0.05) at 18 degrees C than at 28 degrees C in the first experiment but this difference was significant in the second experiment. Rats fed the low protein diet had significantly higher (P > 0.05) heat production than those fed medium or high protein diets. Compared to 28 degrees C, environmental temperature of 18 degrees C caused an increased total and ouabain-sensitive O2 consumption in duodenal mucosa. There was no significant effect of environmental temperature on total and ouabain-sensitive O2 consumption in the liver. However, ouabain-sensitive O2 consumption in liver was significantly higher (P 0.05) when rats were fed a low protein diet compared to the medium or high protein diet. Total and ouabain-sensitive O2 consumption increased in duodenal mucosa of rats fed low level of dietary fibre compared to the medium or high dietary fibre diets. The in vitro results corresponded with the whole animal energy expenditure and O2 consumption in vivo.

Causal Relationships among Technology Acquisition, Absorptive Capacity, and Innovation Performance: Evidence from the Pharmaceutical Industry.

PubMed

Jeon, Jieun; Hong, Suckchul; Ohm, Jay; Yang, Taeyong

2015-01-01

This paper discusses the importance of absorptive capacity in improving a firm's innovation performance. Specifically, we examine firm interaction with the knowledge and capabilities of outside organizations and the effect on the firm's bottom line. We use the impulse-response function of the vector auto-regressive model to gain insight into this relationship by estimating the time required for the effect of each activity level to reach outputs, the spillover effects. We apply this methodology to pharmaceutical firms, which we classify into two sub-groups--large firms and medium and small firms--based on sales. Our results show that the impact of an activity on any other activity is delayed by three years for large firms and by one to two years for small and medium firms.

Tolerance of Bacteria to High Concentrations of NaCl and Glycerol in the Growth Medium

PubMed Central

Marshall, Betty J.; Ohye, D. F.; Christian, J. H. B.

1971-01-01

When compared at similar levels of water activity, glycerol was more inhibitory than sodium chloride to relatively salt-tolerant bacteria and less inhibitory than salt to salt-sensitive species. PMID:5549707

Accumulation of mesalazine pills in the medium ileum in a patient with Crohn´s disease.

PubMed

Martínez Huertas, Carmen; Garcia-Villanova Ruiz, Paloma; Pozo Sánchez, José; Dávila Arias, Cristina

2017-03-01

Crohn´s disease is an inflammatory disease that can involve any portion of the gastrointestinal tract, although terminal ileum is the most commonly affected portion. It is characterized by a transmural and discontinuous distribution pattern, with alternating periods of active disease and remission. We present the case of a 23-year-old patient diagnosed with Crohn´s disease, in treatment with extended release Mesalazine and corticoids. The CT Enterography showed activity signs and a great dilatation of medium ileum with lots of Mesalazine pills accumulated inside. Pill accumulation occurred because of stenosis, which did not let the pills at this level progress to distal ileum, and be absorbed.

Cost-effective medium for the production of mosquito pupicidal lipopeptide from Bacillus subtilis subsp. subtilis (VCRC B471).

PubMed

Bhuvaneswari, S; Manonmani, A M; Geetha, I

2015-03-01

A cyclic lipopeptide (CLP), surfactin produced by a strain of Bacillus subtilis subsp. subtilis (VCRC B471) was found to exhibit mosquitocidal activity. The present study was carried out to enhance the surfactin level using low cost material in the production medium. Two carbon sources, glucose and common sugar, and two nitrogen sources, ammonium nitrate and soya were used in the study. Different concentrations of 'C' and 'N' sources were used in the production medium to enhance the production of surfactin. A new medium (SS7) containing 2% sugar, 6% soya and 0.5% common salt with micronutrients was designed which was found to enhance the production of surfactin. The crude mosquitocidal metabolite (CMM) produced in this medium was 3 g/l which was two times higher than that obtained using synthetic medium NYSM. The LC50 dosage of the CMM to the pupal stages of An. stephensi (2.3 μg/ml) was comparable to that obtained with CMM from the conventional medium. The newly designed cost-effective medium designated as sugar soya medium (SSM) enhanced the production of surfactin and the cost of production was estimated as [symbol: see text] 6 per litre, which is six times lesser than that of the conventional medium. Replacement of sodium chloride with cooking salt further reduced the cost of the medium.

Assessing polyphenols content and antioxidant activity in coffee beans according to origin and the degree of roasting

PubMed

Dybkowska, Ewa; Sadowska, Anna; Rakowska, Rita; Dębowska, Maria; Świderski, Franciszek; Świąder, Katarzyna

The roasting stage constitutes a key component in the manufacturing process of natural coffee because temperature elicits changes in bioactive compounds such as polyphenols and that Maillard-reaction compounds appear, thus affecting the product’s sensory and antioxidant properties. Actual contents of these compounds may depend on which region the coffee is cultivated as well as the extent to which the beans are roasted To determine polyphenols content and antioxidant activity in the ‘Arabica’ coffee type coming from various world regions of its cultivation and which have undergone industrial roasting. Also to establish which coffee, taking into account the degree of roasting (ie. light, medium and strong), is nutritionally the most beneficial The study material was natural coffee beans (100% Arabica) roasted to various degrees, as aforementioned, that had been cultivated in Brazil, Ethiopia, Columbia and India. Polyphenols were measured in the coffee beans by spectrophotometric means based on the Folin-Ciocalteu reaction, whereas antioxidant activity was measured colourimetrically using ABTS+ cat-ionic radicals Polyphenol content and antioxidant activity were found to depend both on the coffee’s origin and degree of roasting. Longer roasting times resulted in greater polyphenol degradation. The highest polyphenol concentrations were found in lightly roasted coffee, ranging 39.27 to 43.0 mg/g, whereas levels in medium and strongly roasted coffee respectively ranged 34.06 to 38.43 mg/g and 29.21 to 36.89 mg/g. Antioxidant activity however significantly rose with the degree of roasting, where strongly roasted coffee had higher such activity than lightly roasted coffee. This can be explained by the formation of Maillard-reaction compounds during roasting, leading then to the formation of antioxidant melanoidin compounds which, to a large extent, compensate for the decrease in polyphenols during roasting Polyphenols levels and antioxidant activities in the studied Arabica coffee beans that had undergone roasting depended on the cultivation region of the world. Longer roasting caused a significant decline in polyphenols compound levels (from 7.3% to 32.1%) in the coffee beans. Antioxidant activities of coffee increased with roasting, despite reduced levels of natural antioxidants. From a nutritional standpoint, the most favoured coffees are those lightly or medium roasted

Study of factors affecting growth and cold acclimation of Vitis callus cultures

DOE Office of Scientific and Technical Information (OSTI.GOV)

Deng, L.

1987-01-01

In vitro grape tissue culture initiation, growth, and cold acclimation were studied. Factors involved were genotypes, media, plant growth regulators, age, light, temperature, antioxidant, clearing and adsorbing agents, sucrose level, osmotic potential, ABA, chilling and freezing treatments. Murashige and Skoog (MS) medium containing 1 ..mu..M 2,4-d + 0.1 uM Ba, MS containing 1 uM 2,4-D, and woody plant medium containing 1 uM 2,4-D + 0.1 uM BA produced abundant callus tissue for most grape genotypes; either WPM or MS containing 1 uM BA stimulated shoot growth in all the 12 genotypes tested. Adding 1 uM abscisic acid (ABA) to themore » B5 medium with 1 uM 2,4-D and 0.5 uM BA enhanced growth and quality of Chancellor callus. /sup 3/H-ABA was taken up actively by callus tissue at 12 days after subculture, but by 20 d this effect disappeared. When /sup 14/C-sucrose was added to the medium. /sup 14/C level of cells reached a plateau after 48 h; this plateau was higher if ABA was also present in the medium. Cells on media containing ABA were larger in size, lighter in color, and more loosely connected.« less

Response surface optimization of medium components for naringinase production from Staphylococcus xylosus MAK2.

PubMed

Puri, Munish; Kaur, Aneet; Singh, Ram Sarup; Singh, Anubhav

2010-09-01

Response surface methodology was used to optimize the fermentation medium for enhancing naringinase production by Staphylococcus xylosus. The first step of this process involved the individual adjustment and optimization of various medium components at shake flask level. Sources of carbon (sucrose) and nitrogen (sodium nitrate), as well as an inducer (naringin) and pH levels were all found to be the important factors significantly affecting naringinase production. In the second step, a 22 full factorial central composite design was applied to determine the optimal levels of each of the significant variables. A second-order polynomial was derived by multiple regression analysis on the experimental data. Using this methodology, the optimum values for the critical components were obtained as follows: sucrose, 10.0%; sodium nitrate, 10.0%; pH 5.6; biomass concentration, 1.58%; and naringin, 0.50% (w/v), respectively. Under optimal conditions, the experimental naringinase production was 8.45 U/mL. The determination coefficients (R(2)) were 0.9908 and 0.9950 for naringinase activity and biomass production, respectively, indicating an adequate degree of reliability in the model.

Evaluation of cellular influences of platinum nanoparticles by stable medium dispersion.

PubMed

Horie, Masanori; Kato, Haruhisa; Endoh, Shigehisa; Fujita, Katsuhide; Nishio, Keiko; Komaba, Lilian Kaede; Fukui, Hiroko; Nakamura, Ayako; Miyauchi, Arisa; Nakazato, Tetsuya; Kinugasa, Shinichi; Yoshida, Yasukazu; Hagihara, Yoshihisa; Morimoto, Yasuo; Iwahashi, Hitoshi

2011-11-01

Platinum nanoparticles have industrial application, for example in catalysis, and are used in consumer products such as cosmetics and supplements. Therefore, among the many nanoparticles, platinum is one of the more accessible nanoparticles for consumers. Most platinum nanoparticles that are used in cosmetics and supplements which have an anti-oxidant activity are modified particles. However, the cellular influences of pristine platinum nanoparticles are still unclear, although it has been reported that platinum nanoparticles induce oxidative stress. In this study, we investigated the cellular influences induced by pure pristine platinum nanoparticles. Platinum nanoparticles of 100% purity were dispersed in a cell culture medium and stable medium dispersion was obtained. The platinum nanoparticle medium dispersion was applied to two kinds of cultured cells, A549 and HaCaT cells, and the cellular influences were examined. Cell viability (MTT assay), cell proliferation (clonogenic assay), apoptosis induction (caspase-3 activity), intracellular ROS level (DCFH assay), and lipid peroxidation level (DPPP assay) were measured as markers of cellular influences. Transmission electron microscope observation showed cellular uptake of platinum nanoparticles. However, the platinum nanoparticles did not drive any markers. It is known that some metal oxide nanoparticles such as NiO and CuO show severe cytotoxicity via metal ion release. Compared with these toxic nanoparticles, the platinum nanoparticles used in this study did not release platinum ions into the culture media. These results suggest that the physically and chemically inactive cellular influences of platinum nanoparticles are small.

The effect of microgravity on the in vitro NK cell function during six International Space Station Missions

NASA Astrophysics Data System (ADS)

Buravkova, L. B.; Grigorieva, V.; Rykova, M. P.

2007-09-01

The level of natural killer (NK) cytotoxic activity was measured during co-cultivation of human lymphocytes and target cells (K-562) in microgravity. Flight experiments were carried out using special instrumentation, the "Fibroblast-1 " cassettes, in the frame of Russian scientific program during six ISS missions. Lymphocyte suspensions from human venous blood were used in experiments during short-term flights on six ISS missions (7-12). Russian space crew members performed the experiments after Soyuz docking. The first step was mixing lymphocytes and3H-labeled K-562 cells and their incubation at 37°C during 24 hs; the second step was filtration of the cell suspension. The frozen medium and filters were analyzed for the cytokine level and cytotoxic activity after landing. It was found that lymphocytes with different basal levels of cytotoxic activity kept the ability of recognizing and lysing malignant cells. In microgravity, cytotoxity increased to 160% of the basal levels. Donor individual features modulated the magnitude of the increase. The measurement of interleukin levels (TNF-α, IL-1, IL-2) in medium showed that synthesis of TNF-α increased during cell co-cultivation in microgravity. The level of IL-2 was very low inflight and ground control samples. The production of IL-1 by lymphocytes decreased after in-flight incubation. The results indicate that microgravity did not disturb the cytotoxic function of immune cells in vitro during 24 h incubation with specific target cells.

Characteristics of phytoplankton in Lake Karachay, a storage reservoir of medium-level radioactive waste.

PubMed

Atamanyuk, Natalia I; Osipov, Denis I; Tryapitsina, Galina A; Deryabina, Larisa V; Stukalov, Pavel M; Ivanov, Ivan A; Pryakhin, Evgeny A

2012-07-01

The status of the phytoplankton community in Lake Karachay, a storage reservoir of liquid medium-level radioactive waste from the Mayak Production Association, Chelyabinsk Region, Russia, is reviewed. In 2010, the concentration of Sr in water of this reservoir was found to be 6.5 × 10(6) Bq L, the concentration of 137Cs was 1.6 × 10(7) Bq L, and total alpha activity amounted to 3.0 × 10(3) Bq L. An increased level of nitrates was observed in the reservoir-4.4 g L. It has been demonstrated that in this reservoir under the conditions of the maximum contamination levels known for aquatic ecosystems in the entire biosphere, a phytoplankton community exists that has a pronounced decline in species diversity, almost to the extent of a monoculture of widely-spread thread eurytopic cyanobacteria Geitlerinema amphibium.

Melatonin enhances lipid production in Monoraphidium sp. QLY-1 under nitrogen deficiency conditions via a multi-level mechanism.

PubMed

Zhao, Yongteng; Li, Dafei; Xu, Jun-Wei; Zhao, Peng; Li, Tao; Ma, Huixian; Yu, Xuya

2018-07-01

In this study, melatonin (MT) promoted lipid accumulation in Monoraphidium sp. QLY-1 under nitrogen deficiency conditions. The lipid accumulation increased 1.22- and 1.36-fold compared with a nitrogen-starved medium and a normal BG-11 medium, respectively. The maximum lipid content was 51.38%. The reactive oxygen species (ROS) level in the presence of melatonin was lower than that in the control group, likely because of the high antioxidant activities. The application of melatonin upregulated the gibberellin acid (GA) production and rbcL and accD expression levels but downregulated the abscisic acid (ABA) content and pepc expression levels. These findings demonstrated that exogenous melatonin could further improve the lipid production in Monoraphidium sp. QLY-1 by regulating antioxidant systems, signalling molecules, and lipid biosynthesis-related gene expression under nitrogen deficiency conditions. Copyright © 2018 Elsevier Ltd. All rights reserved.

Treatment optimization in MS: Canadian MS Working Group updated recommendations.

PubMed

Freedman, Mark S; Selchen, Daniel; Arnold, Douglas L; Prat, Alexandre; Banwell, Brenda; Yeung, Michael; Morgenthau, David; Lapierre, Yves

2013-05-01

The Canadian Multiple Sclerosis Working Group (CMSWG) developed practical recommendations in 2004 to assist clinicians in optimizing the use of disease-modifying therapies (DMT) in patients with relapsing multiple sclerosis. The CMSWG convened to review how disease activity is assessed, propose a more current approach for assessing suboptimal response, and to suggest a scheme for switching or escalating treatment. Practical criteria for relapses, Expanded Disability Status Scale (EDSS) progression and MRI were developed to classify the clinical level of concern as Low, Medium and High. The group concluded that a change in treatment may be considered in any RRMS patient if there is a high level of concern in any one domain (relapses, progression or MRI), a medium level of concern in any two domains, or a low level of concern in all three domains. These recommendations for assessing treatment response should assist clinicians in making more rational choices in their management of relapsing MS patients.

Gaseous oxygen uptake in porous media at different moisture contents and airflow velocities.

PubMed

Sharma, Prabhakar; Poulsen, Tjalfe G; Kalluri, Prasad N V

2009-06-01

The presence and distribution of water in the pore space is a critical factor for flow and transport of gases through unsaturated porous media. The water content also affects the biological activity necessary for treatment of polluted gas streams in biofilters. In this research, microbial activity and quantity of inactive volume in a porous medium as a function of moisture content and gas flow rate were investigated. Yard waste compost was used as a test medium, and oxygen uptake rate measurements were used to quantify microbial activity and effective active compost volume using batch and column flow-through systems. Compost water contents were varied from air-dry to field capacity and gas flows ranged from 0.2 to 2 L x min(-1). The results showed that overall microbial activity and the relative fraction of active compost medium volume increased with airflow velocity for all levels of water content up to a certain flow rate above which the oxygen uptake rate assumed a constant value independent of gas flow. The actual value of the maximum oxygen uptake rate was controlled by the water content. The oxygen uptake rate also increased with increasing water content and reached a maximum between 42 and 48% volumetric water content, above which it decreased, again likely because of formation of inactive zones in the compost medium. Overall, maximum possible oxygen uptake rate as a function of gas flow rate across all water contents and gas flows could be approximated by a linear expression. The relative fraction of active volume also increased with gas flow rate and reached approximately 80% for the highest gas flows used.

Effects of Copper Pollution on the Phenolic Compound Content, Color, and Antioxidant Activity of Wine.

PubMed

Sun, Xiangyu; Ma, Tingting; Han, Luyang; Huang, Weidong; Zhan, Jicheng

2017-05-03

The effects of copper pollution on the polyphenol content, color, and antioxidant activity of wine, as well as correlations among these factors, were investigated. Copper had clear influences on wine polyphenol content. At low copper concentrations, the concentrations of nearly all polyphenols increased, and the antioxidant activity values of the wine also increased. When the copper concentration reached the lowest level of the medium copper range (9.6~16 mg/L), most of the indices also improved. When the copper concentrations reached the latter part of the medium copper range (19.2 and 22.4 mg/L), many of the tested indices began to decrease. Furthermore, when the copper concentration reached the high ranges (32, 64, and 96 mg/L), the polyphenol content, CIELAB color parameters, and antioxidant activity of wine were substantially decreased, indicating the need to control increasing copper content in grape must.

Probing the Molecular Mechanism of Hypericin-Induced Parasite Death Provides Insight into the Role of Spermidine beyond Redox Metabolism in Leishmania donovani

PubMed Central

Singh, Shalini; Sarma, Shyamali; Katiyar, Shashank P.; Das, Mousumi; Bhardwaj, Ruchika; Sundar, Durai

2014-01-01

Hypericin, a natural compound from Hypericum perforatum (St. John's wort), has been identified as a specific inhibitor of Leishmania donovani spermidine synthase (LdSS) using integrated computational and biochemical approaches. Hypericin showed in vitro inhibition of recombinant LdSS enzyme activity. The in vivo estimation of spermidine levels in Leishmania promastigotes after hypericin treatment showed significant decreases in the spermidine pools of the parasites, indicating target specificity of the inhibitor molecule. The inhibitor, hypericin, showed significant antileishmanial activity, and the mode of death showed necrosis-like features. Further, decreased trypanothione levels and increased glutathione levels with elevated reactive oxygen species (ROS) levels were observed after hypericin treatment. Supplementation with trypanothione in the medium with hypericin treatment restored in vivo trypanothione levels and ROS levels but could not prevent necrosis-like death of the parasites. However, supplementation with spermidine in the medium with hypericin treatment restored in vivo spermidine levels and parasite death was prevented to a large extent. The data overall suggest that the parasite death due to spermidine starvation as a result of LdSS inhibition is not related to elevated levels of reactive oxygen species. This suggests the involvement of spermidine in processes other than redox metabolism in Leishmania parasites. Moreover, the work provides a novel scaffold, i.e., hypericin, as a potent antileishmanial molecule. PMID:25313212

Stimulation of d- and l-lactate dehydrogenases transcriptional levels in presence of diammonium hydrogen phosphate resulting to enhanced lactic acid production by Lactobacillus strain.

PubMed

Singhvi, Mamata; Zendo, Takeshi; Iida, Hiroshi; Gokhale, Digambar; Sonomoto, Kenji

2017-12-01

The present study revealed the effect of nitrogen sources on lactic acid production and stimulation of d- and l-lactate dehydrogenases (LDH) of parent Lactobacillus lactis NCIM 2368 and its mutant RM2-24 generated after UV mutagenesis. Both the parent and mutant strains were evaluated for d-lactic acid production in control and modified media. The modified media did not show remarkable effect on lactic acid production in case of parent whereas mutant exhibited significant enhancement in d-lactic acid production along with the appearance of l-lactic acid in the broth. Both LDH activities and specific activities were found to be higher in mutant than the parent strain. These results suggested that the diammonium hydrogen phosphate in modified media triggered the expression of LDH genes leading to enhanced lactic acid production. This observation has been proved by studying the expression levels of d- and l-LDH genes of parent and mutant in control and modified media using quantitative RT-PCR technique. In case of mutant, the transcriptional levels of d-LDH and l-LDH increased ∼17 fold and ∼1.38 fold respectively in modified medium compared to the values obtained with control medium. In case of parent, no significant change in transcriptional levels of d- and l-LDH was found when the cells were grown in either control medium or modified medium. This study suggested that the mutant, RM2-24 has l-LDH gene which is expressed in presence of (NH 4 ) 2 HPO 4 resulting in l-lactic acid production. Co-production of l-lactic acid in d-lactic acid fermentation may be detrimental in the PLA production. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Xylanase production with xylan rich lignocellulosic wastes by a local soil isolate of Trichoderma viride

PubMed Central

Goyal, Meenakshi; Kalra, K.L.; Sareen, V.K.; Soni, G.

2008-01-01

In the present study, cultural and nutritional conditions for enhanced production of xylanase by a local soil isolate of Trichoderma viride, using various lignocellulosic substrates in submerged culture fermentation have been optimized. Of the lignocellulosics used, maize straw was the best inducer followed by jowar straw for xylanase production. The highest activity achieved was between 14 to 17 days of fermentation. A continuous increase in xylanase production was observed with increasing level of lignocellulosics in the medium and highest activity was observed with maize straw at 5% level. Xylanase production with higher levels of lignocellulosics (3 to 5%) of maize, jowar and barseem was found to be higher as compared to that with commercial xylan as carbon source. Sodium nitrate was the best nitrogen source among the six sources used. Maximum xylanase production was achieved with initial medium pH of 3.5–4.0 and incubation temperature of 25ºC.The enzyme preparation was effective in bringing about saccharification of different lignocellulosics. The xylanase production could be further improved by using alkali treated straw as carbon source. PMID:24031262

Osmotic regulation of myo-inositol uptake in primary astrocyte cultures.

PubMed

Isaacks, R E; Bender, A S; Kim, C Y; Prieto, N M; Norenberg, M D

1994-03-01

Uptake of myo-inositol by astrocytes in hypertonic medium (440 mosm/kg H2O) was increased near 3-fold after incubation for 24 hours, which continued for 72 hours, as compared with the uptake by cells cultured in isotonic medium (38 nmoles/mg protein). myo-Inositol uptake by astrocytes cultured in hypotonic medium (180 mosm/kg H2O) for periods up to 72 hours was reduced by 74% to 8 to 10 nmoles/mg protein. Astrocytes incubated in either hypotonic or hypertonic medium for 24 hours and then placed in isotonic medium reversed the initial down- or up-regulation of uptake. Activation of chronic RVD and RVI correlates with regulation of myo-inositol uptake. A 30 to 40 mosm/kg H2O deviation from physiological osmolality can influence myo-inositol homeostasis. The intracellular content of myo-inositol in astrocytes in isotonic medium was 25.6 +/- 1.3 micrograms/mg protein (28 mM). This level of myo-inositol is sufficient for this compound to function as an osmoregulator in primary astrocytes and it is likely to contribute to the maintenance of brain volume.

Active carbon-pools in rhizosphere of wheat (Triticum aestivum L.) grown under elevated atmospheric carbon dioxide concentration in a Typic Haplustept in sub-tropical India.

PubMed

Kant, Pratap C B; Bhadraray, Subhendu; Purakayastha, T J; Jain, Vanita; Pal, Madan; Datta, S C

2007-05-01

Study on active and labile carbon-pools can serve as a clue for soil organic carbon dynamics on exposure to elevated level of CO2. Therefore, an experimental study was conducted in a Typic Haplustept in sub-tropical semi-arid India with wheat grown in open top chambers at ambient (370 micromol mol-1) and elevated (600 micromol mol-1) concentrations of atmospheric CO2. Elevated atmospheric CO2 caused increase in yield and carbon uptake by all plant parts, and their preferential partitioning to root. Increases in fresh root weight, volume and length have also been observed. Relative contribution of medium-sized root to total root length increased at the expense of very fine roots at elevated CO2 level. All active carbon-fractions gained due to elevated atmospheric CO2 concentration, and the order followed their relative labilities. All the C-pools have recorded a significant increase over initial status, and are expected to impart short-to-medium-term effect on soil carbon sequestration.

Preventive effect of Zea mays L. (purple waxy corn) on experimental diabetic cataract.

PubMed

Thiraphatthanavong, Paphaphat; Wattanathorn, Jintanaporn; Muchimapura, Supaporn; Wipawee, Thukham-mee; Wannanon, Panakaporn; Terdthai, Tong-un; Suriharn, Bhalang; Lertrat, Kamol

2014-01-01

Recently, substances possessing antioxidant can prevent cataractogenesis of diabetic cataract. Therefore, this study was carried out to determine the anticataract effect of Zea mays L. (purple waxy corn), a flavonoids rich plant, in experimental diabetic cataract. Enucleated rat lenses were incubated in artificial aqueous humor containing 55 mM glucose with various concentrations of Zea mays L. (purple waxy corn) ranging between 2, 10, and 50 mg/mL at room temperature for 72 h. At the end of the incubation period, the evaluation of lens opacification, MDA level, and the activities of SOD, CAT, GPx, and AR in lens were performed. The results showed that both medium and high doses of extract decreased lens opacity together with the decreased MDA level. In addition, medium dose of extract increased GPx activity while the high dose decreased AR activity. No other significant changes were observed. The purple waxy corn seeds extract is the potential candidate to protect against diabetic cataract. The mechanism of action may occur via the decreased oxidative stress and the suppression of AR. However, further research in vivo is still essential.

Assessment of Microvascular Abnormalities by Nailfold Capillaroscopy in Juvenile Dermatomyositis After Medium- to Long-Term Followup.

PubMed

Barth, Zoltan; Witczak, Birgit N; Flatø, Berit; Koller, Akos; Sjaastad, Ivar; Sanner, Helga

2018-05-01

In juvenile dermatomyositis (DM), microvascular abnormalities, measured by nailfold capillaroscopy (NFC), are common early in the disease course. We aimed to compare the presence of NFC abnormalities in patients with medium- to long-term juvenile DM with that of controls, and to explore associations between NFC abnormalities and disease activity and other disease characteristics. Fifty-eight juvenile DM patients with a median disease duration of 16.8 (range 2-38) years were clinically examined and compared with matched controls. By NFC, we assessed nailfold capillary density (NCD), giant capillaries, scleroderma, and neovascular pattern (defined as scleroderma active or late pattern). NFC was analyzed with researchers blinded to patient/control identity and disease characteristics. We measured disease activity and damage by validated tools, and patients were categorized as having active or inactive disease according to the Paediatric Rheumatology International Trials Organisation criteria. Compared to controls, patients had decreased NCD (mean ± SD 6.4 ± 2.1/mm versus 7.6 ± 0.8/mm; P = 0.001) and showed more abnormality in all other NFC parameters; 36% of patients versus 4% of controls had NCD <6/mm (P < 0.001). Giant capillaries, scleroderma, and neovascular pattern were found in 9%, 84%, and 41% of patients, respectively. Patients with active disease (n = 30) presented more frequently with neovascular pattern than patients with inactive disease (n = 28) (P = 0.041). Decreased NCD and neovascular pattern were associated with higher levels of disease activity and impaired muscle function. After medium- to long-term followup, juvenile DM patients had decreased NCD and, often, neovascular pattern; both were associated with higher levels of disease activity and impaired muscle function. This suggests that NFC can be a biomarker for disease activity in longstanding juvenile DM too. © 2017, American College of Rheumatology.

Formulation and Statistical Optimization of Culture Medium for Improved Production of Antimicrobial Compound by Streptomyces sp. JAJ06

PubMed Central

Arul Jose, Polpass; Sivakala, Kunjukrishnan Kamalakshi; Jebakumar, Solomon Robinson David

2013-01-01

Streptomyces sp. JAJ06 is a seawater-dependent antibiotic producer, previously isolated and characterised from an Indian coastal solar saltern. This paper reports replacement of seawater with a defined salt formulation in production medium and subsequent statistical media optimization to ensure consistent as well as improved antibiotic production by Streptomyces sp. JAJ06. This strain was observed to be proficient to produce antibiotic compound with incorporation of chemically defined sodium-chloride-based salt formulation instead of seawater into the production medium. Plackett-Burman design experiment was applied, and three media constituents, starch, KBr, and CaCO3, were recognised to have significant effect on the antibiotic production of Streptomyces JAJ06 at their individual levels. Subsequently, Response surface methodology with Box-Behnken design was employed to optimize these influencing medium constituents for the improved antibiotic production of Streptomyces sp. JAJ06. A total of 17 experiments were conducted towards the construction of a quadratic model and a second-order polynomial equation. Optimum levels of medium constituents were obtained by analysis of the model and numerical optimization method. When the strain JAJ06 was cultivated in the optimized medium, the antibiotic activity was increased to 173.3 U/mL, 26.8% increase as compared to the original (136.7 U/mL). This study found a useful way to cultivate Streptomyces sp. JAJ06 for enhanced production of antibiotic compound. PMID:24454383

Causal Relationships among Technology Acquisition, Absorptive Capacity, and Innovation Performance: Evidence from the Pharmaceutical Industry

PubMed Central

Jeon, Jieun; Hong, Suckchul; Ohm, Jay; Yang, Taeyong

2015-01-01

This paper discusses the importance of absorptive capacity in improving a firm’s innovation performance. Specifically, we examine firm interaction with the knowledge and capabilities of outside organizations and the effect on the firm’s bottom line. We use the impulse-response function of the vector auto-regressive model to gain insight into this relationship by estimating the time required for the effect of each activity level to reach outputs, the spillover effects. We apply this methodology to pharmaceutical firms, which we classify into two sub-groups – large firms and medium and small firms – based on sales. Our results show that the impact of an activity on any other activity is delayed by three years for large firms and by one to two years for small and medium firms. PMID:26181440

Physiology of Growth and Sporulation in Bacillus cereus I. Effect of Glutamic and Other Amino Acids

PubMed Central

Buono, F.; Testa, R.; Lundgren, D. G.

1966-01-01

Buono, F. (Syracuse University, Syracuse, N.Y.), R. Testa, and D. G. Lundgren. Physiology of growth and sporulation in Bacillus cereus. I. Effect of glutamic and other amino acids. J. Bacteriol. 91:2291–2299. 1966.—Growth and sporulation were studied in Bacillus cereus by use of an active culture technique and a synthetic medium. A high level of glutamic acid (70 mm) was required for optimal growth and glucose oxidation followed by sporulation even though relatively little glutamic acid was consumed (14 mm). Optimal growth occurred with a combination of 14 mm glutamic acid and 56 mm (NH4)2SO4, aspartic acid, or alanine. Ornithine or arginine at 70 mm could replace glutamic acid in the synthetic medium without affecting the normal growth cycle. Glutamic acid was not replaced by any other amino acid, by (NH4)2SO4, or by a combination of either α-ketoglutarate or pyruvate plus (NH4)2SO4. Enzyme assays of cell-free extracts prepared from cells harvested at different times were used to study the metabolism of glutamic acid. Glutamic-oxaloacetic and glutamic-pyruvate transaminases were completely activated (or derepressed) during early stages of sporulation (period of 6 to 8 hr). Alanine dehydrogenase responded in a similar manner, but the levels of this enzyme were much higher throughout the culture cycle. Neither glutamic dehydrogenase nor α-ketoglutarate dehydrogenase was detected. Sporulation in a replacement salts medium was studied with cells harvested at different times from the synthetic medium. Cultures 2 to 6 hr old were unable to sporulate in the replacement salts medium unless glutamic acid (7.0 mm) was present. By the 6th hr, cells were in the early stages of sporulation, showing spore septa development. Cultures 8 hr old sporulated in the replacement salts medium. Other metabolic intermediates able to replace glutamic acid in the replacement salts medium were alanine, aspartic acid, and glutamine at equimolar concentrations. Also, ammonium ions in combination with pyruvic, oxaloacetic, α-ketoglutaric, or fumaric acid replaced glutamic acid. The likely role of these metabolites is discussed. PMID:4957615

Intermittent hydrostatic pressure inhibits matrix metalloproteinase and pro-inflammatory mediator release from human osteoarthritic chondrocytes in vitro.

PubMed

Trindade, Michael C D; Shida, Jun-ichi; Ikenoue, Takashi; Lee, Mel S; Lin, Eric Y; Yaszay, Burt; Yerby, Scott; Goodman, Stuart B; Schurman, David J; Smith, R Lane

2004-09-01

This study tested the hypothesis that intermittent hydrostatic pressure applied to human osteoarthritic chondrocytes modulates matrix metalloproteinase and pro-inflammatory mediator release in vitro. Human osteoarthritic articular chondrocytes were isolated and cultured as primary high-density monolayers. For testing, chondrocyte cultures were transferred to serum-free medium and maintained without loading or with exposure to intermittent hydrostatic pressure (IHP) at 10 MPa at a frequency of 1 Hz for periods of 6, 12 and 24 h. Levels of matrix metalloproteinase-2, -9 (MMP-2, -9), tissue inhibitor of metalloproteinase-1 (TIMP-1), and the pro-inflammatory mediators, interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1), released into the culture medium were assessed by ELISA. Matrix metalloproteinase activity was confirmed by zymographic analysis. In the absence of IHP, levels of MMP-2, TIMP-1, IL-6, and MCP-1 in the chondrocyte culture medium increased in a time-dependent manner. Application of IHP decreased MMP-2 levels at all time periods tested, relative to unloaded control cultures maintained for the same time periods. Although 84/82 kDa bands were faintly detectable by zymography, MMP-9 levels were not quantifiable in medium from loaded or unloaded cultures by ELISA. TIMP-1 levels were not altered in response to IHP at any time period tested. IL-6 and MCP-1 levels decreased in cultures exposed to IHP at 12 and 24 h, relative to unloaded control cultures maintained for the same time periods. IHP decreased release of MMP-2, IL-6 and MCP-1 by osteoarthritic chondrocytes in vitro suggesting that pressure influences cartilage stability by modulating chondrocyte expression of these degradative and pro-inflammatory proteins in vivo.

Cytotoxic and hemolytic effects of Tritrichomonas foetus on mammalian cells.

PubMed Central

Burgess, D E; Knoblock, K F; Daugherty, T; Robertson, N P

1990-01-01

Geographically distinct lines of Tritrichomonas foetus were assayed for their ability to cause cytotoxicity in nucleated mammalian cells and lysis of bovine erythrocytes. T. foetus was highly cytotoxic toward a human cervical cell line (HeLa) and early bovine lymphosarcoma (BL-3) but displayed low levels of cytotoxicity against African green monkey kidney (Vero) cells. In addition to variation in the extent of cytotoxicity toward different targets, differences in the levels of cytotoxicity in the same nucleated target occurred with different parasite lines. Whole T. foetus, unfractionated whole-cell extracts, and parasite-conditioned medium (RPMI 1640 without serum) all caused lysis of bovine erythrocytes. Lytic activity in the conditioned medium was substantially reduced by repeated freezing and thawing or heating to 90 degrees C for 30 min. Damage of mammalian target cells by live T. foetus could be reduced by the presence of protease inhibitors; however, such inhibitors did not diminish the lytic effects of conditioned medium. These results suggested that proteolytic enzymes were necessary for the lytic mechanism of the live parasites but were not required once lytic factors were released into the parasite-conditioned medium. They further suggested that the lytic molecules were either proteins or had proteinaceous components. Images PMID:2228233

An analysis of long and medium-haul air passenger demand, volume 1

NASA Technical Reports Server (NTRS)

Eriksen, S. E.

1978-01-01

A basic model was developed which is a two equation pair econometric system in which air passenger demand and airline level-of-service are the endogenous variables. The model aims to identify the relationship between each of these two variables and its determining factors, and to identify the interaction of demand and level-of-service with each other. The selected variable for the measure of air passenger traffic activity in a given pair market is defined as the number of passengers in a given time that originate in one region and fly to the other region for purposes other than to make a connection to a third region. For medium and long haul markets, the model seems to perform better for larger markets. This is due to a specification problem regarding the route structure variable. In larger markets, a greater percentage of nonlocal passengers are accounted for by this variable. Comparing the estimated fare elasticities of long and medium haul markets, it appears that air transportation demand is more price elastic in longer haul markets. Long haul markets demand will saturate with a fewer number of departures than will demand in medium haul markets.

Determination of lytic enzyme activities of indigenous Trichoderma isolates from Pakistan.

PubMed

Asad, Saeed Ahmad; Tabassum, Ayesha; Hameed, Abdul; Hassan, Fayyaz Ul; Afzal, Aftab; Khan, Sabaz Ali; Ahmed, Rafiq; Shahzad, Muhammad

2015-01-01

This study investigated lytic enzyme activities in three indigenous Trichoderma strains namely, Trichoderma asperellum, Trichoderma harzianum and Trichoderma sp. Native Trichoderma strains and a virulent strain of Rhizoctonia solani isolated from infected bean plants were also included in the study. Enzyme activities were determined by measuring sugar reduction by dinitrosalicylic acid (DNS) method using suitable substrates. The antagonists were cultured in minimal salt medium with the following modifications: medium A (1 g of glucose), medium B (0.5 g of glucose + 0.5 g of deactivated R. solani mycelia), medium C (1.0 g of deactivated respective antagonist mycelium) and medium D (1 g of deactivated R. solani mycelia). T asperellum showed presence of higher amounts of chitinases, β-1, 3-glucanases and xylanases in extracellular protein extracts from medium D as compared to medium A. While, the higher activities of glucosidases and endoglucanses were shown in medium D extracts by T. harzianum. β-glucosidase activities were lower compared with other enzymes; however, activities of the extracts of medium D were significantly different. T. asperellum exhibited maximum inhibition (97.7%). On the other hand, Trichoderma sp. did not show any effect on mycelia growth of R. solani on crude extract.

Determination of lytic enzyme activities of indigenous Trichoderma isolates from Pakistan

PubMed Central

Asad, Saeed Ahmad; Tabassum, Ayesha; Hameed, Abdul; Hassan, Fayyaz ul; Afzal, Aftab; Khan, Sabaz Ali; Ahmed, Rafiq; Shahzad, Muhammad

2015-01-01

Abstract This study investigated lytic enzyme activities in three indigenous Trichoderma strains namely, Trichoderma asperellum, Trichoderma harzianum and Trichoderma sp. Native Trichoderma strains and a virulent strain of Rhizoctonia solani isolated from infected bean plants were also included in the study. Enzyme activities were determined by measuring sugar reduction by dinitrosalicylic acid (DNS) method using suitable substrates. The antagonists were cultured in minimal salt medium with the following modifications: medium A (1 g of glucose), medium B (0.5 g of glucose + 0.5 g of deactivated R. solani mycelia), medium C (1.0 g of deactivated respective antagonist mycelium) and medium D (1 g of deactivated R. solani mycelia). T asperellum showed presence of higher amounts of chitinases, β-1, 3-glucanases and xylanases in extracellular protein extracts from medium D as compared to medium A. While, the higher activities of glucosidases and endoglucanses were shown in medium D extracts by T. harzianum. β-glucosidase activities were lower compared with other enzymes; however, activities of the extracts of medium D were significantly different. T. asperellum exhibited maximum inhibition (97.7%). On the other hand, Trichoderma sp. did not show any effect on mycelia growth of R. solani on crude extract. PMID:26691463

Chitin Synthesis in Saccharomyces cerevisiae in Response to Supplementation of Growth Medium with Glucosamine and Cell Wall Stress

PubMed Central

Bulik, Dorota A.; Olczak, Mariusz; Lucero, Hector A.; Osmond, Barbara C.; Robbins, Phillips W.; Specht, Charles A.

2003-01-01

In Saccharomyces cerevisiae most chitin is synthesized by Chs3p, which deposits chitin in the lateral cell wall and in the bud-neck region during cell division. We have recently found that addition of glucosamine (GlcN) to the growth medium leads to a three- to fourfold increase in cell wall chitin levels. We compared this result to the increases in cellular chitin levels associated with cell wall stress and with treatment of yeast with mating pheromone. Since all three phenomena lead to increases in precursors of chitin, we hypothesized that chitin synthesis is at least in part directly regulated by the size of this pool. This hypothesis was strengthened by our finding that addition of GlcN to the growth medium causes a rapid increase in chitin synthesis without any pronounced change in the expression of more than 6,000 genes monitored with Affymetrix gene expression chips. In other studies we found that the specific activity of Chs3p is higher in the total membrane fractions from cells grown in GlcN and from mutants with weakened cell walls. Sucrose gradient analysis shows that Chs3p is present in an inactive form in what may be Golgi compartments but as an active enzyme in other intracellular membrane-bound vesicles, as well as in the plasma membrane. We conclude that Chs3p-dependent chitin synthesis in S. cerevisiae is regulated both by the levels of intermediates of the UDP-GlcNAc biosynthetic pathway and by an increase in the activity of the enzyme in the plasma membrane. PMID:14555471

Influence of petroleum hydrocarbon contamination on microalgae and microbial activities in a long-term contaminated soil.

PubMed

Megharaj, M; Singleton, I; McClure, N C; Naidu, R

2000-05-01

Petroleum hydrocarbons are widespread environmental pollutants. Although biodegradation of petroleum hydrocarbons has been the subject of numerous investigations, information on their toxicity to microorganisms in soil is limited, with virtually no work conducted on soil algae. We carried out a screening experiment for total petroleum hydrocarbons (TPH) and their toxicity to soil algal populations, microbial biomass, and soil enzymes (dehydrogenase and urease) in a long-term TPH-polluted site with reference to an adjacent unpolluted site. Microbial biomass, soil enzyme activity, and microalgae declined in medium to high-level (5,200-21,430 mg kg(-1) soil) TPH-polluted soils, whereas low-level (<2,120 mg kg(-1) soil) pollution stimulated the algal populations and showed no effect on microbial biomass and enzymes. However, inhibition of all the tested parameters was more severe in soil considered to have medium-level pollution than in soils that were highly polluted. This result could not be explained by chemical analysis alone. Of particular interest was an observed shift in the species composition of algae in polluted soils with elimination of sensitive species in the medium to high polluted soils. Also, an algal growth inhibition test carried out using aqueous eluates prepared from polluted soils supported these results. Given the sensitivity of algae to synthetic pollutants, alteration in the algal species composition can serve as a useful bioindicator of pollution. The results of this experiment suggest that chemical analysis alone is not adequate for toxicological estimations and should be used in conjunction with bioassays. Furthermore, changes in species composition of algae proved to be more sensitive than microbial biomass and soil enzyme activity measurements.

Cutinase production by Fusarium oxysporum in liquid medium using central composite design.

PubMed

Pio, Tatiana Fontes; Macedo, Gabriela Alves

2008-01-01

The objective of the present study was to measure the production of cutinase by Fusarium oxysporum in the presence of several carbon and nitrogen sources (glycides, fatty acids and oils, and several organic and inorganic nitrogen sources), trying to find a cost-effective substitute for cutin in the culture medium as an inducer of cutinase production. The results were evaluated by the Tukey test, and flaxseed oil was found to give the best results as a cutinase inducer. The authors optimized the composition of the growth medium employing response surface methodology. The experimental results were fitted to a second-order polynomial model at a 95% level of significance (p < 0.05). The greatest cutinolytic activity was obtained in a liquid mineral medium supplemented with flaxseed oil, showing an increase in enzymatic activity from 11 to 22.68 U/mL after 48 h of fermentation. A CCD study of the fermentation conditions was carried out, and the best production of cutinase was registered with the use of 30 degrees C and 100 rpm. These results support the use of flaxseed oil as a substitute for cutin, which is difficult and expensive to obtain, for the production of cutinase in a larger scale.

Optimization of media composition for Nattokinase production by Bacillus subtilis using response surface methodology.

PubMed

Deepak, V; Kalishwaralal, K; Ramkumarpandian, S; Babu, S Venkatesh; Senthilkumar, S R; Sangiliyandi, G

2008-11-01

Response surface methodology and central composite rotary design (CCRD) was employed to optimize a fermentation medium for the production of Nattokinase by Bacillus subtilis at pH 7.5. The four variables involved in this study were Glucose, Peptone, CaCl2, and MgSO4. The statistical analysis of the results showed that, in the range studied; only peptone had a significant effect on Nattokinase production. The optimized medium containing (%) Glucose: 1, Peptone: 5.5, MgSO4: 0.2 and CaCl2: 0.5 resulted in 2-fold increased level of Nattokinase (3194.25U/ml) production compared to initial level (1599.09U/ml) after 10h of fermentation. Nattokinase production was checked with fibrinolytic activity.

Activity of blood coagulation and fibrinolysis during and after hydroxyethyl starch (HES) colloidal volume replacement.

PubMed

Omar, M N; Shouk, T A; Khaleq, M A

1999-06-01

To examine the effect of medium molecular weight hydroxyethyl starch on protein C levels and the changes in the activation state of blood platelets, coagulation and fibrinolyis during and after 5 day of its infusion. Fifty male patients (mean age: 47 years, range 45-50 years) who required prostatectomy for benign prostatic hyperplasia were divided into two equal groups. One group was given 15 mL/kg body weight (mean volume 1000 mL +/- 100 mL) of 6% hydroxyethyl starch (HES) 200/0.5, the other received an equal volume of 5% human albumin during the operation. Blood samples were collected immediately before infusion (baseline values) and at 20, 40, 60, 90, 240, and 480 min after the infusion started then daily for the next 5 days postoperatively. Hematocrit, factor VIII:C, thrombin-antithrombin III complex; the anticoagulant protein C levels; the fibrinolytic parameters tissue type plasminogen activator (t-PA), and the fibrinolytic product D-Dimer and the platelet aggregation activity were measured. The data obtained did not detect any significant differences between HES and human albumin in the plasma levels of thrombin-antithrombin III complex, protein C, tissue-type plasminogen activator and the fibrin split products D-Dimer. Factor VIII:C and platelet aggregation were significantly lower in the hydroxyethyl starch group in comparison with albumin. Baseline values were attained postoperatively for factor VIII:C and platelet aggregation by the first and fifth days, respectively. The lowering effect of medium molecular weight hydroxyethyl starch on factor VIII:C would not be attributed to increased proteolytic activity of protein C on this coagulation cofactor because there is a nonsignificant change in protein C levels.

The inhibitory effects of sorbate and benzoate against Clostridium perfringens type A isolates.

PubMed

Alnoman, Maryam; Udompijitkul, Pathima; Paredes-Sabja, Daniel; Sarker, Mahfuzur R

2015-06-01

This study evaluated the inhibitory effects of sorbate and benzoate against Clostridium perfringens type A food poisoning (FP) and non-food-borne (NFB) disease isolates. No significant inhibition of germination of spores of both FP and NFB isolates was observed in rich medium (pH 7.0) supplemented with permissive level of sodium sorbate (0.3% ≈ 0.13 mM undissociated sorbic acid) or sodium benzoate (0.1% ≈ 0.01 mM undissociated benzoic acid) used in foods. However, these levels of sorbate and benzoate effectively arrested outgrowth of germinated C. perfringens spores in rich medium. Lowering the pH of the medium increases the inhibitory effects of sorbate and benzoate against germination of spores of NFB isolates, and outgrowth of spores of both FP and NFB isolates. Furthermore, sorbate and benzoate inhibited vegetative growth of C. perfringens isolates. However, the permissible levels of these organic salts could not control the growth of C. perfringens spores in chicken meat stored under extremely abusive conditions. In summary, although sorbate and benzoate showed inhibitory activities against C. perfringens in the rich medium, no such effect was observed in cooked chicken meat. Therefore, caution should be taken when applying these organic salts into meat products to reduce or eliminate C. perfringens spores. Copyright © 2014 Elsevier Ltd. All rights reserved.

The Impact of the Roast Levels of Coffee Extracts on their Potential Anticancer Activities.

PubMed

Mojica, Benigno E; Fong, Lisa E; Biju, Denny; Muharram, Alfeah; Davis, Isabel M; Vela, Klarisse O; Rios, Diana; Osorio-Camacena, Elena; Kaur, Baljit; Rojas, Sebastian M; Forester, Sarah C

2018-04-01

Coffee is one of the most widely consumed beverages in the world and contains numerous phytochemicals that are beneficial to consumer health. The phytochemical profile of coffee, however, can be affected by the roast level. In this study, we compared the effect of roasting level on the growth inhibitory activity of HT-29 (colon) and SCC-25 (oral) cancer cell lines. The different roasting stages selected for this study were green, cinnamon/blonde, city/medium, full city/medium-dark, and full city plus/dark. Cancer cells were treated with various concentrations of coffee extracts for 72 hr. Cell viability was quantified using the thiazolyl blue tetrazolium bromide assay. It was found that the lighter roast extracts, Cinnamon in particular, reduced cell growth more than darker roast extracts. The Cinnamon extract had the greatest amount of total phenolic content and antioxidant activity. Relative levels of gallic, caffeic, and chlorogenic acid in the extracts were also compared. The Cinnamon coffee extract had the highest levels of gallic and caffeic acids, which have both been widely-regarded as bioactive phytochemicals. In conclusion, the consumption of lighter roasted coffee, may contribute to the prevention of certain types of cancer such as oral and colon. Chemical compounds in coffee may reduce the risk for certain types of cancers. These compounds may be particularly abundant in lighter roasted coffee. Therefore, lighter roasted coffee could contribute to the prevention of cancer through a healthy diet. © 2018 Institute of Food Technologists®.

Rhythmic expression of DEC2 protein in vitro and in vivo.

PubMed

Sato, Fuyuki; Muragaki, Yasuteru; Kawamoto, Takeshi; Fujimoto, Katsumi; Kato, Yukio; Zhang, Yanping

2016-06-01

Basic helix-loop-helix (bHLH) transcription factor DEC2 (bHLHE41/Sharp1) is one of the clock genes that show a circadian rhythm in various tissues. DEC2 regulates differentiation, sleep length, tumor cell invasion and apoptosis. Although studies have been conducted on the rhythmic expression of DEC2 mRNA in various tissues, the precise molecular mechanism of DEC2 expression is poorly understood. In the present study, we examined whether DEC2 protein had a rhythmic expression. Western blot analysis for DEC2 protein revealed a rhythmic expression in mouse liver, lung and muscle and in MCF-7 and U2OS cells. In addition, AMP-activated protein kinase (AMPK) activity (phosphorylation of AMPK) in mouse embryonic fibroblasts (MEFs) exhibited a rhythmic expression under the condition of medium change or glucose-depleted medium. However, the rhythmic expression of DEC2 in MEF gradually decreased in time under these conditions. The medium change affected the levels of DEC2 protein and phosphorylation of AMPK. In addition, the levels of DEC2 protein showed a rhythmic expression in vivo and in MCF-7 and U2OS cells. The results showed that the phosphorylation of AMPK immunoreactivity was strongly detected in the liver and lung of DEC2 knockout mice compared with that of wild-type mice. These results may provide new insights into rhythmic expression and the regulation between DEC2 protein and AMPK activity.

Return to recreational sports activity after anterior cruciate ligament reconstruction: a one- to six-year follow-up study.

PubMed

Rodríguez-Roiz, Juan M; Caballero, Miguel; Ares, Oscar; Sastre, Sergi; Lozano, Luis; Popescu, Dragos

2015-08-01

The aim of this study was to evaluate mid-term return to recreational sport in general population and identify factors related with sports return. Retrospective evaluation of 99 recreational sports players (Tegner score of 6) with ACL arthroscopic reconstructions with hamstring autograft, between 2006-2011. 74 male and 25 female with middle age of 30 years (14-52). We made a questionnaire focused on sports level before injury and after surgery, and different scales: Lysholm, Tegner Activity Level, IKDC and a Likert scale for quantify their motivation for return to sports. With a medium follow-up of 36 months, 90 patients (91.9 %) had returned to recreational sport. 51 (51.52 %) had returned to sports at the same level, and these are those with lower BMI (average 23), higher IKDC and Lysholm scores (p < 0.01) and also they believe that sport is an important activity. Only 9 % of patients left sports. They were principally male athletes (88 %), with higher medium age (32), lesser time between injury and surgery (22 months), higher BMI (26), in comparison with athletes that return to sports. The results suggest good mid-term return to recreational sports in general population. The following factors had a statistically significant influence on the return to sports activity: type of sport, sex and functional state of the operated knee (IKDC-Lysholm). Psychological and social factors may have a fundamental influence on return to sports activity.

Metabolomics Guides Rational Development of a Simplified Cell Culture Medium for Drug Screening against Trypanosoma brucei

PubMed Central

Creek, Darren J.; Nijagal, Brunda; Kim, Dong-Hyun; Rojas, Federico; Matthews, Keith R.

2013-01-01

In vitro culture methods underpin many experimental approaches to biology and drug discovery. The modification of established cell culture methods to make them more biologically relevant or to optimize growth is traditionally a laborious task. Emerging metabolomic technology enables the rapid evaluation of intra- and extracellular metabolites and can be applied to the rational development of cell culture media. In this study, untargeted semiquantitative and targeted quantitative metabolomic analyses of fresh and spent media revealed the major nutritional requirements for the growth of bloodstream form Trypanosoma brucei. The standard culture medium (HMI11) contained unnecessarily high concentrations of 32 nutrients that were subsequently removed to make the concentrations more closely resemble those normally found in blood. Our new medium, Creek's minimal medium (CMM), supports in vitro growth equivalent to that in HMI11 and causes no significant perturbation of metabolite levels for 94% of the detected metabolome (<3-fold change; α = 0.05). Importantly, improved sensitivity was observed for drug activity studies in whole-cell phenotypic screenings and in the metabolomic mode of action assays. Four-hundred-fold 50% inhibitory concentration decreases were observed for pentamidine and methotrexate, suggesting inhibition of activity by nutrients present in HMI11. CMM is suitable for routine cell culture and offers important advantages for metabolomic studies and drug activity screening. PMID:23571546

RESONATORS. MODES: Modes of a plano - spherical laser resonator with the Gaussian gain distribution of the active medium

NASA Astrophysics Data System (ADS)

Malyutin, A. A.

2007-03-01

Modes of a laser with plano-spherical degenerate and nondegenerate resonators are calculated upon diode pumping producing the Gaussian gain distribution in the active medium. Axially symmetric and off-axis pumpings are considered. It is shown that in the first case the lowest Hermite-Gaussian mode is excited with the largest weight both in the degenerate and nondegenerate resonator if the pump level is sufficiently high or the characteristic size wg of the amplifying region greatly exceeds the mode radius w0. The high-order Ince-Gaussian modes are excited upon weak off-axis pumping in the nondegenerate resonator both in the absence and presence of the symmetry of the gain distribution with respect to the resonator axis. It is found that when the level of off-axis symmetric pumping of the resonator is high enough, modes with the parameters of the TEM00 mode periodically propagating over a closed path in the resonator can exist. The explanation of this effect is given.

Matrix Metalloproteinase-20 Over-Expression Is Detrimental to Enamel Development: A Mus musculus Model

PubMed Central

Shin, Masashi; Hu, Yuanyuan; Tye, Coralee E.; Guan, Xiaomu; Deagle, Craig C.; Antone, Jerry V.; Smith, Charles E.; Simmer, James P.; Bartlett, John D.

2014-01-01

Background Matrix metalloproteinase-20 (Mmp20) ablated mice have enamel that is thin and soft with an abnormal rod pattern that abrades from the underlying dentin. We asked if introduction of transgenes expressing Mmp20 would revert this Mmp20 null phenotype back to normal. Unexpectedly, for transgenes expressing medium or high levels of Mmp20, we found opposite enamel phenotypes depending on the genetic background (Mmp20−/− or Mmp20+/+) in which the transgenes were expressed. Methodology/Principal Findings Amelx-promoter-Mmp20 transgenic founder mouse lines were assessed for transgene expression and those expressing low, medium or high levels of Mmp20 were selected for breeding into the Mmp20 null background. Regardless of expression level, each transgene brought the null enamel back to full thickness. However, the high and medium expressing Mmp20 transgenes in the Mmp20 null background had significantly harder more mineralized enamel than did the low transgene expresser. Strikingly, when the high and medium expressing Mmp20 transgenes were present in the wild-type background, the enamel was significantly less well mineralized than normal. Protein gel analysis of enamel matrix proteins from the high and medium expressing transgenes present in the wild-type background demonstrated that greater than normal amounts of cleavage products and smaller quantities of higher molecular weight proteins were present within their enamel matrices. Conclusions/Significance Mmp20 expression levels must be within a specific range for normal enamel development to occur. Creation of a normally thick enamel layer may occur over a wider range of Mmp20 expression levels, but acquisition of normal enamel hardness has a narrower range. Since over-expression of Mmp20 results in decreased enamel hardness, this suggests that a balance exists between cleaved and full-length enamel matrix proteins that are essential for formation of a properly hardened enamel layer. It also suggests that few feedback controls are present in the enamel matrix to prevent excessive MMP20 activity. PMID:24466234

Requirement of a Relatively High Threshold Level of Mg2+ for Cell Growth of a Rhizoplane Bacterium, Sphingomonas yanoikuyae EC-S001

PubMed Central

Hoo, Henny; Hashidoko, Yasuyuki; Islam, Md. Tofazzal; Tahara, Satoshi

2004-01-01

Mg2+ is one of the essential elements for bacterial cell growth. The presence of the magnesium cation (Mg2+) in various concentrations often affects cell growth restoration in plant-associating bacteria. This study attempted to determine whether Mg2+ levels in Sphingomonas yanoikuyae EC-S001 affected cell growth restoration in the host plant and what the threshold level is. S. yanoikuyae EC-S001, isolated from the rhizoplane of spinach seedlings grown from surface-sterilized seeds under aseptic conditions, displayed uniform dispersion and attachment throughout the rhizoplane and phylloplane of the host seedlings. S. yanoikuyae EC-S001 did not grow in potato-dextrose broth medium but grew well in an aqueous extract of spinach leaves. Chemical investigation of the growth factor in the spinach leaf extract led to identification of the active principle as the magnesium cation. A concentration of ca. 0.10 mM Mg2+ or more allowed S. yanoikuyae EC-S001 to grow in potato-dextrose broth medium. Some saprophytic and/or diazotrophic bacteria used in our experiment were found to have diverse threshold levels for their Mg2+ requirements. For example, Burkholderia cepacia EC-K014, originally isolated from the rhizoplane of a Melastoma sp., could grow even in Mg2+-free Hoagland's no. 2 medium with saccharose and glutamine (HSG medium) and requires a trace level of Mg2+ for its growth. In contrast, S. yanoikuyae EC-S001, together with Bacillus subtilis IFO12113, showed the most drastic restoring responses to subsequent addition of 0.98 mM Mg2+ to Mg2+-free HSG medium. Our studies concluded that Mg2+ is more than just the essential trace element needed for cell growth restoration in S. yanoikuyae EC-S001 and that certain nonculturable bacteria may require a higher concentration of Mg2+ or another specific essential element for their growth. PMID:15345402

Acidic pH stimulates the production of the angiogenic CXC chemokine, CXCL8 (interleukin-8), in human adult mesenchymal stem cells via the extracellular signal-regulated kinase, p38 mitogen-activated protein kinase, and NF-kappaB pathways.

PubMed

Bischoff, David S; Zhu, Jian-Hua; Makhijani, Nalini S; Yamaguchi, Dean T

2008-07-01

Blood vessel injury results in limited oxygen tension and diffusion leading to hypoxia, increased anaerobic metabolism, and elevated production of acidic metabolites that cannot be easily removed due to the reduced blood flow. Therefore, an acidic extracellular pH occurs in the local microenvironment of disrupted bone. The potential role of acidic pH and glu-leu-arg (ELR(+)) CXC chemokines in early events in bone repair was studied in human mesenchymal stem cells (hMSCs) treated with medium of decreasing pH (7.4, 7.0, 6.7, and 6.4). The cells showed a reciprocal increase in CXCL8 (interleukin-8, IL-8) mRNA levels as extracellular pH decreased. At pH 6.4, CXCL8 mRNA was induced >60x in comparison to levels at pH 7.4. hMSCs treated with osteogenic medium (OGM) also showed an increase in CXCL8 mRNA with decreasing pH; although, at a lower level than that seen in cells grown in non-OGM. CXCL8 protein was secreted into the medium at all pHs with maximal induction at pH 6.7. Inhibition of the G-protein-coupled receptor alpha, G(alphai), suppressed CXCL8 levels in response to acidic pH; whereas phospholipase C inhibition had no effect on CXCL8. The use of specific mitogen-activated protein kinase (MAPK) signal transduction inhibitors indicated that the pH-dependent increase in CXCL8 mRNA is due to activation of ERK and p38 pathways. The JNK pathway was not involved. NF-kappaB inhibition resulted in a decrease in CXCL8 levels in hMSCs grown in non-OGM. However, OGM-differentiated hMSCs showed an increase in CXCL8 levels when treated with the NF-kappaB inhibitor PDTC, a pyrrolidine derivative of dithiocarbamate. 2008 Wiley-Liss, Inc.

Comparison of sugar molecule decomposition through glucose and fructose: a high-level quantum chemical study.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Assary, R. S.; Curtiss, L. A.; MSD)

Efficient chemical conversion of biomass is essential to produce sustainable energy and industrial chemicals. Industrial level conversion of glucose to useful chemicals, such as furfural, hydroxymethylfurfural, and levulinic acid, is a major step in the biomass conversion but is difficult because of the formation of undesired products and side reactions. To understand the molecular level reaction mechanisms involved in the decomposition of glucose and fructose, we have carried out high-level quantum chemical calculations [Gaussian-4 (G4) theory]. Selective 1,2-dehydration, keto-enol tautomerization, isomerization, retro-aldol condensation, and hydride shifts of glucose and fructose molecules were investigated. Detailed kinetic and thermodynamic analyses indicate that,more » for acyclic glucose and fructose molecules, the dehydration and isomerization require larger activation barriers compared to the retro-aldol reaction at 298 K in neutral medium. The retro-aldol reaction results in the formation of C2 and C4 species from glucose and C3 species from fructose. The formation of the most stable C3 species, dihydroxyacetone from fructose, is thermodynamically downhill. The 1,3-hydride shift leads to the cleavage of the C-C bond in the acyclic species; however, the enthalpy of activation is significantly higher (50-55 kcal/mol) than that of the retro-aldol reaction (38 kcal/mol) mainly because of the sterically hindered distorted four-membered transition state compared to the hexa-membered transition state in the retro-aldol reaction. Both tautomerization and dehydration are catalyzed by a water molecule in aqueous medium; however, water has little effect on the retro-aldol reaction. Isomerization of glucose to fructose and glyceraldehyde to dihydroxyacetone proceeds through hydride shifts that require an activation enthalpy of about 40 kcal/mol at 298 K in water medium. This investigation maps out accurate energetics of the decomposition of glucose and fructose molecules that is needed to help find more efficient catalysts for the conversion of hexose to useful chemicals.« less

Association between Municipal Health Promotion Volunteers’ Health Literacy and Their Level of Outreach Activities in Japan

PubMed Central

Taguchi, Atsuko; Murayama, Hiroshi; Murashima, Sachiyo

2016-01-01

Objectives To explore the association between health literacy and levels of three types of core activities among health promotion volunteers (developing a healthy lifestyle, outreach to family, and outreach to community members). Study Design A cross-sectional, anonymous, self-administered postal survey of registered health promotion volunteers in the Konan area in Shiga Prefecture in Japan, conducted in January 2010. The study sample was 575 registered health promotion volunteers. Methods The survey collected data on health literacy, gender, age, education, self-rated health, perceptions about the volunteer organization, and perceptions of recognition in the community. The level of engagement in health promotion activities was measured by the extent to which the participants engaged in seven healthy behaviors and promoted them to family members and the community. The authors compared the health literacy level and other characteristics of the participants by core health promotion activities, using a chi-squared test, to examine the associations between demographic and other variables and the three core activities (healthy lifestyle, outreach to family, and outreach to community).Logistic regression analysis was conducted to examine the association between the degree to which the volunteers engaged in core activities (“healthy lifestyle,” “outreach to family,” “outreach to community”) and the levels of health literacy (low, medium, high) among health promotion volunteers, controlling for the effects of age, gender, health condition, education which may also have an impact on volunteers’ outreach activities. Results Four hundred and fifty-four questionnaires were returned, a 79.0% response rate. Excluding 16 cases with missing values on health literacy or the degree of health promotion activities, 438 research subjects were included in the analysis (valid response rate: 76.2%). Health literacy and a few demographic and other characteristics of the volunteers were associated with the three core health promotion activities. In bivariate analyses, active participation in the core activities was more prevalent among older volunteers (p<0.001 for all three activities). Self-rated health condition was associated with both outreach to family (p = 0.018) and community (p = 0.046). Years of experience as volunteer and perception of being recognized in the community also had statistically significant association with outreach to the community (p<0.001). In multiple logistic regression, those with higher level of health literacy were more likely than others to actively engage in outreach to family (OR = 1.70, 95% CI 1.03 to 2.80; OR = 1.76, 95%CI 1.04 to 3.00 for medium and high, respectively) and outreach to community (OR = 2.26, 95%CI 1.34 to 3.83; OR = 2.61 95%CI 1.49 to 4.58 for medium and high, respectively). Perception of being recognized in the community also had a statistically significant and positive impact on outreach to the community (OR = 1.52, 95%CI 1.17 to 1.99). Conclusions Volunteers with higher health literacy were more likely to actively engage in outreach to family and outreach to community. Providing educational programs to improve volunteers’ health literacy may facilitate their work. PMID:27736942

Association between Municipal Health Promotion Volunteers' Health Literacy and Their Level of Outreach Activities in Japan.

PubMed

Taguchi, Atsuko; Murayama, Hiroshi; Murashima, Sachiyo

2016-01-01

To explore the association between health literacy and levels of three types of core activities among health promotion volunteers (developing a healthy lifestyle, outreach to family, and outreach to community members). A cross-sectional, anonymous, self-administered postal survey of registered health promotion volunteers in the Konan area in Shiga Prefecture in Japan, conducted in January 2010. The study sample was 575 registered health promotion volunteers. The survey collected data on health literacy, gender, age, education, self-rated health, perceptions about the volunteer organization, and perceptions of recognition in the community. The level of engagement in health promotion activities was measured by the extent to which the participants engaged in seven healthy behaviors and promoted them to family members and the community. The authors compared the health literacy level and other characteristics of the participants by core health promotion activities, using a chi-squared test, to examine the associations between demographic and other variables and the three core activities (healthy lifestyle, outreach to family, and outreach to community).Logistic regression analysis was conducted to examine the association between the degree to which the volunteers engaged in core activities ("healthy lifestyle," "outreach to family," "outreach to community") and the levels of health literacy (low, medium, high) among health promotion volunteers, controlling for the effects of age, gender, health condition, education which may also have an impact on volunteers' outreach activities. Four hundred and fifty-four questionnaires were returned, a 79.0% response rate. Excluding 16 cases with missing values on health literacy or the degree of health promotion activities, 438 research subjects were included in the analysis (valid response rate: 76.2%). Health literacy and a few demographic and other characteristics of the volunteers were associated with the three core health promotion activities. In bivariate analyses, active participation in the core activities was more prevalent among older volunteers (p<0.001 for all three activities). Self-rated health condition was associated with both outreach to family (p = 0.018) and community (p = 0.046). Years of experience as volunteer and perception of being recognized in the community also had statistically significant association with outreach to the community (p<0.001). In multiple logistic regression, those with higher level of health literacy were more likely than others to actively engage in outreach to family (OR = 1.70, 95% CI 1.03 to 2.80; OR = 1.76, 95%CI 1.04 to 3.00 for medium and high, respectively) and outreach to community (OR = 2.26, 95%CI 1.34 to 3.83; OR = 2.61 95%CI 1.49 to 4.58 for medium and high, respectively). Perception of being recognized in the community also had a statistically significant and positive impact on outreach to the community (OR = 1.52, 95%CI 1.17 to 1.99). Volunteers with higher health literacy were more likely to actively engage in outreach to family and outreach to community. Providing educational programs to improve volunteers' health literacy may facilitate their work.

Substrate-Induced Transcriptional Activation of the MoCel7C Cellulase Gene Is Associated with Methylation of Histone H3 at Lysine 4 in the Rice Blast Fungus Magnaporthe oryzae

PubMed Central

Vu, Ba Van; Pham, Kieu Thi Minh

2013-01-01

The mechanisms involved in substrate-dependent regulation of a Magnaporthe oryzae gene encoding a cellulase which we designate MoCel7C (MGG_14954) were investigated. The levels of MoCel7C transcript were dramatically increased more than 1,000-fold, 16 to 24 h after transfer to a medium containing 2% carboxymethylcellulose (CMC), while levels were very low or undetectable in conventional rich medium. Green fluorescent protein reporter assays showed that the MoCel7C promoter was activated by cello-oligosaccharides larger than a pentamer. CMC-induced activation of the MoCel7C promoter was suppressed by glucose and cellobiose. Chromatin immunoprecipitation assays revealed that histone H3 methylation on lysine 4 (H3K4) at the MoCel7C locus was associated with activation of the gene by CMC. Consistently, CMC-induced MoCel7C gene activation was drastically diminished in a knockout (KO) mutant of the MoSET1 gene, which encodes a histone lysine methyltransferase that catalyzes H3K4 methylation in M. oryzae. Interestingly, however, MoCel7C transcript levels under noninducing conditions were significantly increased in the MoSET1 KO mutant, suggesting that MoSET1 directly or indirectly plays a role in both activation and suppression of the MoCel7C gene in response to environmental signals. In addition, gene expression and silencing vectors using the MoCel7C promoter were constructed. PMID:23995923

Efficient Third-Order Distributed Feedback Laser with Enhanced Beam Pattern

NASA Technical Reports Server (NTRS)

Hu, Qing (Inventor); Lee, Alan Wei Min (Inventor); Kao, Tsung-Yu (Inventor)

2015-01-01

A third-order distributed feedback laser has an active medium disposed on a substrate as a linear array of segments having a series of periodically spaced interstices therebetween and a first conductive layer disposed on a surface of the active medium on each of the segments and along a strip from each of the segments to a conductive electrical contact pad for application of current along a path including the active medium. Upon application of a current through the active medium, the active medium functions as an optical waveguide, and there is established an alternating electric field, at a THz frequency, both in the active medium and emerging from the interstices. Spacing of adjacent segments is approximately half of a wavelength of the THz frequency in free space or an odd integral multiple thereof, so that the linear array has a coherence length greater than the length of the linear array.

Medium-Chain Fatty Acids Improve Cognitive Function in Intensively Treated Type 1 Diabetic Patients and Support In Vitro Synaptic Transmission During Acute Hypoglycemia

PubMed Central

Page, Kathleen A.; Williamson, Anne; Yu, Namyi; McNay, Ewan C.; Dzuira, James; McCrimmon, Rory J.; Sherwin, Robert S.

2009-01-01

OBJECTIVE We examined whether ingestion of medium-chain triglycerides could improve cognition during hypoglycemia in subjects with intensively treated type 1 diabetes and assessed potential underlying mechanisms by testing the effect of β-hydroxybutyrate and octanoate on rat hippocampal synaptic transmission during exposure to low glucose. RESEARCH DESIGN AND METHODS A total of 11 intensively treated type 1 diabetic subjects participated in stepped hyperinsulinemic- (2 mU · kg−1 · min−1) euglycemic- (glucose ∼5.5 mmol/l) hypoglycemic (glucose ∼2.8 mmol/l) clamp studies. During two separate sessions, they randomly received either medium-chain triglycerides or placebo drinks and performed a battery of cognitive tests. In vitro rat hippocampal slice preparations were used to assess the ability of β-hydroxybutyrate and octanoate to support neuronal activity when glucose levels are reduced. RESULTS Hypoglycemia impaired cognitive performance in tests of verbal memory, digit symbol coding, digit span backwards, and map searching. Ingestion of medium-chain triglycerides reversed these effects. Medium-chain triglycerides also produced higher free fatty acids and β-hydroxybutyrate levels compared with placebo. However, the increase in catecholamines and symptoms during hypoglycemia was not altered. In hippocampal slices β-hydroxybutyrate supported synaptic transmission under low-glucose conditions, whereas octanoate could not. Nevertheless, octanoate improved the rate of recovery of synaptic function upon restoration of control glucose concentrations. CONCLUSIONS Medium-chain triglyceride ingestion improves cognition without adversely affecting adrenergic or symptomatic responses to hypoglycemia in intensively treated type 1 diabetic subjects. Medium-chain triglycerides offer the therapeutic advantage of preserving brain function under hypoglycemic conditions without causing deleterious hyperglycemia. PMID:19223595

Inhibition of acetylcholinesterase in guppies (Poecilia reticulata) by chlorpyrifos at sublethal concentrations: Methodological aspects

DOE Office of Scientific and Technical Information (OSTI.GOV)

van der Wel, H.; Welling, W.

1989-04-01

Acetylcholinesterase activity is a potential biochemical indicator of toxic stress in fish and a sensitive parameter for testing water for the presence of organophosphates. A number of methodological aspects regarding the determination of the in vivo effect of chlorpyrifos on acetylcholinesterase in guppies have been investigated. It was found that with acetylthiocholine as a substrate, the contribution of pseudocholinesterase to the total cholinesterase activity can be neglected. Protection of acetylcholinesterase of guppies exposed to chlorpyrifos from additional, artifactual in vitro enzyme inhibition during homogenization is necessary. Very low concentrations of acetone in the exposure medium, resulting from dilution of themore » stock solution of chlorpyrifos in acetone, can result in large decreases in the oxygen content of this medium. This may affect the uptake rate of the toxic compound and, thereby, cholinesterase inhibition. Very low, sublethal concentrations of chlorpyrifos result in high inhibition levels of acetylcholinesterase (80-90%) in guppies within 2 weeks of continuous exposure. Recovery of the enzyme activity occurs after the exposed animals are kept in clean medium for 4 days, but the rate of recovery is considerably lower than the rate of inhibition.« less

Extracellular production of an intact and biologically active human growth hormone by the Bacillus brevis system.

PubMed

Kajino, T; Saito, Y; Asami, O; Yamada, Y; Hirai, M; Udata, S

1997-10-01

The characteristic features of the Bacillus brevis system are very high productivity of heterologous proteins and very low extracellular protease activity. However, degradation of some heterologous proteins, especially mammalian proteins, can be observed and resulted in a lowering of protein productivity. By using a mutant expressing low levels of proteases and the addition of EDTA to the medium, intact human growth hormone (hGH) was successfully produced with the B. brevis system. Signal peptide modification with higher basicity in the amino terminal region and higher hydrophobicity in the middle region brought about a twelve-fold increase in hGH production. The hGH yield was further elevated to 240 mg L-1 by optimization of culture conditions. Thus, biologically active and mature hGH can be efficiently produced directly in the medium with the B. brevis system.

The influence of different submerged cultivation conditions on mycelial biomass and protease production by Lentinus citrinus Walleyn et Rammeloo DPUA 1535 (Agaricomycetideae).

PubMed

Kirsch, Larissa de Souza; Pinto, Ana Carolina dos Santos; Porto, Tatiana Souza; Porto, Ana Lúcia Figueiredo; Teixeira, Maria Francisca Simas

2011-01-01

The influence of different carbon and nitrogen sources, pH of the culture medium, and temperature and period of cultivation on mycelial biomass production and protease activity by Lentinus citrinus DPUA 1535 were investigated in submerged culture. A 2(5) full factorial design with three central points was employed, and the results showed that at a significance level of 95% only nitrogen source and temperature were statistically significant for mycelial biomass production. On the other hand, for protease activity all factors and some interactions were significant, and the temperature and nitrogen source had the most significant effect. The best condition for mycelial biomass production (5.76 mg mL(-1)) and protease activity (32.3 U mL(-1)) was obtained in medium formulated with 0.5% soluble starch, 0.2% gelatin, pH 7.0, 25 degrees C, in 5 days.

Ambulatory Physical Activity Performance in Youth With Cerebral Palsy and Youth Who Are Developing Typically

PubMed Central

Bjornson, Kristie F; Belza, Basia; Kartin, Deborah; Logsdon, Rebecca; McLaughlin, John F

2007-01-01

Background and Purpose Assessment of walking activity in youth with cerebral palsy (CP) has traditionally been “capacity-based.” The purpose of this study was to describe the day-to-day ambulatory activity “performance” of youth with CP compared with youth who were developing typically. Subjects Eighty-one youth with CP, aged 10 to 13 years, who were categorized as being in Gross Motor Function Classification System (GMFCS) levels I to III and 30 age-matched youth who were developing typically were recruited. Methods Using a cross-sectional design, participants wore the StepWatch monitor for 7 days while documenting average daily total step counts, percentage of all time active, ratio of medium to low activity levels, and percentage of time at high activity levels. Results The youth with CP demonstrated significantly lower levels of all outcomes than the comparison group. Discussion and Conclusion Daily walking activity and variability decreased as functional walking level (GMFCS level) decreased. Ambulatory activity performance within the context of the daily life for youth with CP appears valid and feasible as an outcome for mobility interventions in CP. PMID:17244693

Use of an in vivo titration method to study a global regulator: effect of varying Lrp levels on expression of gltBDF in Escherichia coli.

PubMed

Borst, D W; Blumenthal, R M; Matthews, R G

1996-12-01

Most studies of global regulatory proteins are performed in vitro or involve phenotypic comparisons between wild-type and mutant strains. We report the use of strains in which the gene for the leucine-responsive regulatory protein (lrp) is transcribed from isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible promoters for the purpose of continuously varying the in vivo concentration of Lrp. To obtain a broad range of Lrp concentrations, strains were employed that contained the lrp fusion either in the chromosome (I. C. Blomfield, P. J. Calie, K. J. Eberhardt, M. S. McClain, and B. I. Eisenstein, J. Bacteriol. 175:27-36, 1993) or on a multicopy plasmid. Western blot (immunoblot) analysis with polyclonal antiserum to Lrp confirmed that Lrp levels could be varied more than 70-fold by growing the strains in glucose minimal 3-(N-morpholino)propanesulfonic acid (MOPS) medium containing different amounts of IPTG. Expression of an Lrp-regulated gltB::lacZ operon fusion was measured over this range of Lrp concentrations. beta-Galactosidase activity rose with increasing Lrp levels up to the level of Lrp found in wild-type strains, at which point expression is maximal. The presence of leucine in the medium increased the level of Lrp necessary to achieve half-maximal expression of the gltB::lacZ fusion, as predicted by earlier in vitro studies (B. R. Ernsting, J. W. Denninger, R. M. Blumenthal, and R. G. Matthews, J. Bacteriol. 175:7160-7169, 1993). Interestingly, levels of Lrp greater than those in wild-type cells interfered with activation of gltB::lacZ expression. The growth rate of cultures correlated with the intracellular Lrp concentration: levels of Lrp either lower or higher than wild-type levels resulted in significantly slower growth rates. Thus, the level of Lrp in the cell appears to be optimal for rapid growth in minimal medium, and the gltBDF control region is designed to give maximal expression at this Lrp level.

Dual behavior of N-acetylcysteine during ethanol-induced oxidative stress in embryonic chick brains.

PubMed

Bauer, Alison K; Fitzgerald, Mary; Ladzinski, Adam T; Lenhart Sherman, Sydney; Maddock, Benjamin H; Norr, Zoe M; Miller, Robert R

2017-10-01

Ethanol (EtOH) causes oxidative stress in embryos. Because N-acetylcysteine (NAC) failures and successes in ameliorating EtOH-induced oxidative stress have been reported, the objective was to determine if exogenous NAC ameliorated EtOH-induced oxidative stress within embryonic chick brains. Control eggs were injected with approximately 25 µl of water on day 0, 1, and 2 of development (E 0-2 ). Experimental eggs were injected with dosages of either 3.0 mmol EtOH/kg egg; 747 µmol NAC/kg egg; 3.0 mmol EtOH and 747 µmol NAC/kg egg; 1000 µmol NAC/kg egg; or 3.0 mmol EtOH and 1000 µmol NAC/kg during the first 3 days of development (E 0-2 ). At 11 days of development (E 11 ; late embryogenesis), brains were harvested and subsequently assayed for oxidative stress markers including the loss of long-chain membrane polyunsaturated fatty acids (PUFAs); the accumulation of lipid hydroperoxides (LPO); decreased glutathione (GSH) and glutathione/glutathione disulfide (GSSG) levels; and decreased glutathione peroxidase (GPx) activities. EtOH (3 mmol/kg egg), medium NAC (747 µmol/kg egg), and EtOH and medium NAC promoted oxidative stress. These treatments caused decreased brain membrane long-chain PUFAs; increased LPO levels; decreased GSH levels and GSH/GSSG levels; and decreased Se-dependent GPx activities. High NAC dosages (1000 µmol/kg egg) attenuated EtOH-induced oxidative stress within EtOH and high NAC-treated chick brains. Exogenous EtOH and/or medium NAC propagated oxidative stress. Meanwhile, high NAC ameliorated EtOH-induced oxidative stress.

The effect of pre-injury physical fitness on the initial severity and recovery from whiplash injury, at six-month follow-up.

PubMed

Geldman, Mark; Moore, Ann; Cheek, Liz

2008-04-01

To evaluate the effect of pre-injury physical fitness on the initial severity and recovery of motor vehicle-induced neck injury (whiplash injury). A quantitative experimental design using both retrospective and prospective data. Metropolitan Police physiotherapy and rehabilitation department in the UK. One-hundred and two patients with neck pain following whiplash injury. Patients were divided into three groups based on pre-injury physical fitness (low, medium and high). Recovery was compared between the three groups initially then again at three and six months. Three measurement scales were used: the Neck Disability Index, the Problem Percentage, and the Physical Activity Scale. Pre-injury physical fitness had a marked effect on recovery at three and six months, with the medium and high fitness groups having significantly better recovery than the low fitness group. At three months the Neck Disability Index score for the low fitness group was 12 compared with 7 and 7.5 for the medium and high fitness groups respectively (P = 0.009). At six months the Neck Disability Index score was 9 for the low fitness group compared with 0 and 3 for the medium and high fitness groups (P = 0.002). In addition, the return to work rate was almost twice as high for individuals with medium/high fitness. Early recovery from whiplash injury was significantly more likely for individuals with medium to high levels of pre-injury physical fitness than for individuals with low levels of pre-injury physical fitness.

[Possibilities and limits of sleep recovery for night-nursing workers].

PubMed

de Medeiros, Soraya Maria; de Macêdo, Maria Lúcia Azevedo Ferreira; de Oliveira, Jonas Sâmi Albuquerque; Ribeiro, Laiane Medeiros

2009-03-01

The present study had the objective of investigating the possibilities/limits of sleep recovery in women workers with medium level in nursing that develop their activities in night shifts in a first-aid clinic at a public hospital in Natal, Rio Grande do Norte, Brazil. This was an analytic study, with a qualitative approach which used the technique of thematic oral history Fifteen women workers with medium level in nursing were interviewed. The following empirical categories were defined: night shift work and night sleep and night rest at work environment Physiological and psychological sufferings are evident on their speech as well as what the lack of sleep causes in health and personal life of these women workers, making the overload of work clear and also the sociocultural imperfections of feminine gender

Metabolic studies of mammalian cells by 31P-NMR using a continuous perfusion technique.

PubMed

Knop, R H; Chen, C W; Mitchell, J B; Russo, A; McPherson, S; Cohen, J S

1984-07-20

Levels of ATP and Pi in metabolically active Chinese hamster lung fibroblasts were monitored noninvasively by 31P-NMR over many hours and under a variety of conditions. The cells were embedded in a matrix of agarose gel in the form of fine threads which were continuously perfused in a standard NMR tube. The small diameter of the thread allows rapid diffusion of metabolites and drugs into the cells. The changes in ATP and Pi levels were followed as a function of time in response to perfusion with a glucose-containing medium, with isotonic saline and with a medium containing 2,4-dinitrophenol, an uncoupler of oxidative phosphorylation. This gel-thread perfusion method should enable routine NMR studies of cellular metabolism, and may have other potential biological applications.

[Influence of activation and blockade of NMDA receptors on extinction of passive avoidance response in mice with different levels of anxiety].

PubMed

Tomilenko, R A; Dubrovina, N I

2006-03-01

Influence of agonist (D-cycloserine) and antagonist (dizocilpine) N-methyl-D-aspartate receptors on learning and extinction of passive avoidance response in medium-, high-, and low-anxious mice was studied. In medium-anxious mice, D-cycloserine (30 mg/kg) although not changing learning accelerated development of extinction, whereas dizocilpine (0.15 mg/kg), while impairing passive avoidance learning, detained the extinction. In high-anxious mice with good retrieval of memory trace and absence of extinction, D-cycloserine was ineffective, whereas dizocilpine reduced learning and promoted retention of memory trace retrieval at the generated level on extinction. In low-anxious mice, D-cycloserine impaired learning and accelerated extinction, whereas dizocilpine completely blocked learning and retention of passive avoidance response.

Method for surface plasmon amplification by stimulated emission of radiation (SPASER)

DOEpatents

Stockman, Mark I [Atlanta, GA; Bergman, David J [Ramat Hasharon, IL

2011-09-13

A nanostructure is used to generate a highly localized nanoscale optical field. The field is excited using surface plasmon amplification by stimulated emission of radiation (SPASER). The SPASER radiation consists of surface plasmons that undergo stimulated emission, but in contrast to photons can be localized within a nanoscale region. A SPASER can incorporate an active medium formed by two-level emitters, excited by an energy source, such as an optical, electrical, or chemical energy source. The active medium may be quantum dots, which transfer excitation energy by radiationless transitions to a resonant nanosystem that can play the same role as a laser cavity in a conventional laser. The transitions are stimulated by the surface plasmons in the nanostructure, causing the buildup of a macroscopic number of surface plasmons in a single mode.

Surface plasmon amplification by stimulated emission of radiation (SPASER)

DOEpatents

Stockman, Mark I [Atlanta, GA; Bergman, David J [Ramat Hasharon, IL

2009-08-04

A nanostructure is used to generate a highly localized nanoscale optical field. The field is excited using surface plasmon amplification by stimulated emission of radiation (SPASER). The SPASER radiation consists of surface plasmons that undergo stimulated emission, but in contrast to photons can be localized within a nanoscale region. A SPASER can incorporate an active medium formed by two-level emitters, excited by an energy source, such as an optical, electrical, or chemical energy source. The active medium may be quantum dots, which transfer excitation energy by radiationless transitions to a resonant nanosystem that can play the same role as a laser cavity in a conventional laser. The transitions are stimulated by the surface plasmons in the nanostructure, causing the buildup of a macroscopic number of surface plasmons in a single mode.

Preventive Effect of Zea mays L. (Purple Waxy Corn) on Experimental Diabetic Cataract

PubMed Central

Thiraphatthanavong, Paphaphat; Wattanathorn, Jintanaporn; Muchimapura, Supaporn; Thukham-mee, Wipawee; Wannanon, Panakaporn; Tong-un, Terdthai; Suriharn, Bhalang; Lertrat, Kamol

2014-01-01

Recently, substances possessing antioxidant can prevent cataractogenesis of diabetic cataract. Therefore, this study was carried out to determine the anticataract effect of Zea mays L. (purple waxy corn), a flavonoids rich plant, in experimental diabetic cataract. Enucleated rat lenses were incubated in artificial aqueous humor containing 55 mM glucose with various concentrations of Zea mays L. (purple waxy corn) ranging between 2, 10, and 50 mg/mL at room temperature for 72 h. At the end of the incubation period, the evaluation of lens opacification, MDA level, and the activities of SOD, CAT, GPx, and AR in lens were performed. The results showed that both medium and high doses of extract decreased lens opacity together with the decreased MDA level. In addition, medium dose of extract increased GPx activity while the high dose decreased AR activity. No other significant changes were observed. The purple waxy corn seeds extract is the potential candidate to protect against diabetic cataract. The mechanism of action may occur via the decreased oxidative stress and the suppression of AR. However, further research in vivo is still essential. PMID:24527449

Changes of microbial activities and soil aggregation in rhizosphere soil of lettuce plants by drought and the possible influence of inoculation with AM fungi and/or PGPR

NASA Astrophysics Data System (ADS)

Kohler, J.; Caravaca, F.; Roldán, A.

2009-04-01

The effect of different arbuscular mycorrhizal (AM) fungi, Glomus intraradices (Schenk & Smith) or Glomus mosseae (Nicol & Gerd.) Gerd. & Trappe, and plant growth-promoting rhizobacteria (PGPR) (Pseudomonas mendocina Palleroni), alone or in combination, on structural stability and microbial activity in the rhizosphere soil of Lactuca sativa L. was assessed under well-watered conditions and two levels of drought. Desiccation caused an increase in aggregate stability and water-soluble and total carbohydrates but there were no significant differences among treated soils and the control soil. The glomalin-related soil protein (GRSP) levels in both the <2 mm and 0.2-4 mm soil fractions increased with medium water stress, whereas under severe water stress they did not differ with respect to those of well-watered soils. The values of GRSP in soils inoculated with PGPR and AM fungi were higher than in the control or fertilised soil under well-watered and severe-drought conditions, while under medium-drought conditions all soils showed similar GRSP values. Soils inoculated with AM fungi and PGPR generally presented higher dehydrogenase and phosphatase activities than the control soil, independent of the water regime.

Modification of mitochondrial function, cytoplasmic lipid content and cryosensitivity of bovine embryos by resveratrol.

PubMed

Abe, Takahito; Kawahara-Miki, Ryouka; Hara, Tomotaka; Noguchi, Tatsuo; Hayashi, Takeshi; Shirasuna, Koumei; Kuwayama, Takehito; Iwata, Hisataka

2017-10-18

Resveratrol is a potent activator of NAD-dependent deacetyltransferase sirtuin-1 (SIRT1) and affects lipid metabolism and ATP generation in somatic cells. In the present study, the effects of supplementing culture medium with resveratrol on lipid metabolism, ATP generation, and cryosensitivity of bovine in vitro produced embryos were investigated. Bovine early cleaved-stage embryos were cultured in medium containing 0 or 0.5 µM resveratrol for 1 or 5 days. Resveratrol treatment for both 1 day and 5 days increased the expression levels of SIRT1 and phosphorylated AMP-activated protein kinase (pAMPK) in the embryos. Furthermore, resveratrol treatment was effective to increase ATP generation and reduce lipid content of the embryos. The effects of resveratrol treatment were diminished by the SIRT1 inhibitor "EX527", and the reduced lipid content was reversed by treatment with etomoxir (a potent inhibitor of beta-oxidation). Blastocysts developed after resveratrol treatment showed low levels reactive oxygen species and increased cryotolerance. These results demonstrate that resveratrol improves in vitro development of bovine embryos, while reducing cytoplasmic lipid content through activation of beta-oxidation, thereby effective for production of bovine blastocysts with enhanced cryotolerance.

Amplification and gas-dynamic parameters of the active oxygen-iodine medium produced by an ejector nozzle unit

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zagidullin, M V; Nikolaev, V D; Svistun, M I

2001-08-31

The gain, the temperature, and the absolute velocity of the supersonic active oxygen-iodine medium produced by an ejector nozzle unit were determined by the technique of high-resolution diode laser spectroscopy. The gain in the active medium is formed at less than 44 mm from the nozzle unit for an absolute flow velocity {nu} {approx} 600 m s{sup -1}. Upon dilution of oxygen by primary nitrogen in the ratio of 1 : 6.9, the gain of the active medium amounts to 7x10{sup -3} cm{sup -1}, the temperature of the active medium to 200 K, the absolute flow velocity to 580 mmore » s{sup -1}, and the pressure to 58 Torr. As the dilution is increased to 1 : 13.5, the gain reduces to 4.5x10{sup -3} cm{sup -1}, the temperature lowers to 180 K, the velocity of the active medium increases to 615 m s{sup -1}, and the pressure increases to 88 Torr. The increase in the initial content of water vapour in the oxygen flow results in an increase in the temperature and a decrease in the gain of the active medium. (active media)« less

Calcium protects Trifolium repens L. seedlings against cadmium stress.

PubMed

Wang, Chang Quan; Song, Heng

2009-09-01

The effect of calcium (Ca(2+)) on Trifolium repens L. seedlings subjected to cadmium (Cd(2+)) stress was studied by investigating plant growth and changes in activity of antioxidative enzymes. Physiological analysis was carried out on seedlings cultured for 2 weeks on half-strength Hoagland medium with Cd(2+) concentrations of 0, 400 and 600 microM, and on corresponding medium supplied with CaCl(2) (5 mM). Exposure to increasing Cd(2+) reduced the fresh weight of the upper part (stems + leaves) of the seedlings more strongly than that of the root system. In both parts of T. repens seedlings H(2)O(2) level and lipid peroxidation increased. In the upper part, Cd(2+) exposure led to a significant decrease in the activity of superoxide dismutase, catalase and glutathione peroxidase and an increase in ascorbate peroxidase activity. In contrast, the roots showed an increase in the activity of antioxidative enzymes under Cd(2+) stress. Ca(2+) addition to medium reduced the Cd(2+) accumulation, and considerably reversed the Cd(2+)-induced decrease in fresh mass as well as the changes in lipid peroxidation in the both parts of T. repens seedlings. Ca(2+) application diminished the Cd(2+) effect on the activity of antioxidative enzymes in the upper part, even though it did not significantly affect these enzymes in the roots. So the possible mechanisms for the action of Ca(2+) in Cd(2+) stress were considered to reduce Cd(2+) accumulation, alleviate lipid peroxidation and promote activity of antioxidative enzymes.

Production of pectate lyases and cellulases by Chryseomonas luteola strain MFCL0 depends on the growth temperature and the nature of the culture medium: evidence for two critical temperatures.

PubMed

Laurent, P; Buchon, L; Guespin-Michel, J F; Orange, N

2000-04-01

Several extracellular enzymes that are responsible for plant tissue maceration were detected in culture supernatant of the psychrotrophic bacterium Chryseomonas luteola MFCL0. Isoelectrofocusing experiments showed that pectate lyase (PL) activity resulted from the cumulative action of three major isoenzymes, designated PLI, PLII, and PLIII. Cellulolytic activity was also detected in culture supernatants. These enzymes exhibited different behaviors with respect to growth temperature. PLII was not regulated by temperature, whereas PLI and PLIII were regulated similarly by growth temperature. Maximal levels of PLI and PLIII were produced at 14 degrees C when cells were grown in polygalacturonate-containing synthetic medium and at around 20 to 24 degrees C in nutrient broth. In contrast, thermoregulation of cellulolytic activity production differed from thermoregulation of PL. The level of cellulolytic activity was low in all media at temperatures up to 20 degrees C, and then it increased dramatically until the temperature was 28 degrees C, which is the optimal temperature for growth of C. luteola. Previously, we defined the critical temperature by using the modified Arrhenius equation to characterize bacterial behavior. This approach consists of monitoring changes in the maximal specific growth rate as a function of temperature. Our most striking result was the finding that the temperature at which maximum levels of PLI and PLIII were produced in two different media was the same as the critical temperature for growth observed in these two media.

Chitosan inhibits enterotoxigenic Clostridium perfringens type A in growth medium and chicken meat.

PubMed

Alnoman, Maryam; Udompijitkul, Pathima; Sarker, Mahfuzur R

2017-06-01

Clostridium perfringens is a spore-forming bacterium and a major cause of bacterial food-borne illness. In this study, we evaluated the inhibitory effects of chitosan against spore germination, spore outgrowth and vegetative growth of C. perfringens food poisoning (FP) isolates. Chitosan of differing molecular weights inhibited germination of spores of all tested FP isolates in a KCl germinant solution containing 0.1 mg/ml chitosan at pH 4.5. However, higher level (0.25 mg/ml) of chitosan was required to effectively arrest outgrowth of the germinated C. perfringens spores in Tripticase-yeast extract-glucose (TGY) medium. Furthermore, chitosan (1.0 mg/ml) was bacteriostatic against vegetative cells of C. perfringens in TGY medium. Although chitosan showed strong inhibitory activities against C. perfringens in laboratory medium, higher levels (2.0 mg/g) were required to achieve similar inhibition of spores inoculated into chicken meat. In summary, the inhibitory effects of chitosan against C. perfringens FP isolates was concentration dependent, and no major difference was observed when using different molecule weight chitosan as an inhibitor. Our results contribute to a better understanding on the potential application of chitosan in cooked meat products to control C. perfringens-associated disease. Copyright © 2016 Elsevier Ltd. All rights reserved.

Cr(VI) reduction from contaminated soils by Aspergillus sp. N2 and Penicillium sp. N3 isolated from chromium deposits.

PubMed

Fukuda, Tsubasa; Ishino, Yasuhiro; Ogawa, Akane; Tsutsumi, Kadzuyo; Morita, Hiroshi

2008-10-01

Aspergillus sp. N2 and Penicillium sp. N3 are chromate-resistant filamentous fungi that were isolated from Cr(VI) contaminated soil based on their ability to decrease hexavalent chromium levels in the growth medium. After 120 h of growth in a medium containing 50 ppm Cr(VI) at near neutral pH, Aspergillus sp. N2 reduced the Cr(VI) concentration by about 75%. Penicillium sp. N3 was able to reduce the Cr(VI) concentration by only 35%. However, Penicillium sp. N3 reduced the Cr(VI) concentration in the medium by 93% under acidic conditions. Interestingly, the presence of Cu(II) enhanced the Cr(VI) reducing ability of Aspergillus sp. N2 and Penicillium sp. N3 at near neutral pH. Aspergillus sp. N2 and Penicillium sp. N3 reduced the Cr(VI) concentration in the growth medium to a virtually undetectable level within 120 h. For both Aspergillus sp. N2 and Penicillium sp. N3, mycelial seed cultures were more efficient at Cr(VI) reduction than conidium seed cultures. The mechanisms of Cr(VI) reduction in Aspergillus sp. N2 and Penicillium sp. N3 were enzymatic reduction and sorption to mycelia. Enzymatic activity contributed significantly to Cr(VI) reduction. Aspergillus sp. N2 and Penicillium sp. N3 reduced the levels of Cr(VI) in polluted soil samples, suggesting that these strains might be useful for cleaning up chromium-contaminated sites.

Metabolic active-high density VERO cell cultures on microcarriers following apoptosis prevention by galactose/glutamine feeding.

PubMed

Mendonça, Ronaldo Z; Arrózio, Sara J; Antoniazzi, Marta M; Ferreira, Jorge M C; Pereira, Carlos A

2002-07-17

The control of cell death occurring in high density cultures performed in bioreactors is an important factor in production processes. In this work, medium nutrient removal or feeding was used to determine at which extension apoptosis could be, respectively, involved or prevented in VERO cell cultures on microcarriers. Glutamine and galactose present in the VERO cell culture medium was consumed after, respectively, 6 and 12 days of culture. Kinetics studies showed that fresh medium replacement and, to some extent, galactose or glutamine depleted-fresh medium replacement provided a nutritional environment, allowing the VERO cell cultures to attain high densities. Galactose was shown to be a more critical nutrient when cultures reached a high density. In agreement with that, VERO cell cultures supplemented with galactose and/or glutamine were shown to confirm previous findings and, again at high densities, galactose was shown to be a critical nutrient for VERO cell growth. These observations also indicated that in VERO cell cultures, for feeding purposes, the glutamine could be replaced by galactose. The inverse was not true and led, at high densities, to a decrease of cell viability. In the absence of glutamine and galactose, apoptosis was observed in VERO cell cultures by cytofluorometry, Acridine orange staining or light and electron microscopy, reaching high levels when compared to cultures performed with complete medium. VERO cells apoptosis process could be prevented by the galactose and/or glutamine feeding and, at high densities, galactose was more efficient in protecting the cultures. These cultures, prevented from apoptosis, were shown to synthesize high levels of measles virus following infection. Our data show that apoptosis prevention by glutamine/galactose feeding, led to high productive and metabolic active VERO cell cultures, as indicated by the high cell density obtained and the virus multiplication leading to higher virus titers.

The utility of injury narratives for prioritising future prevention activities in small and medium enterprises in rural Vietnam.

PubMed

Marucci-Wellman, Helen; Willetts, Joanna L; Diep, Nguyen Bich; Binh, Ta Thi Tuyet

2013-04-01

Injuries are a leading cause of work-related disability and death in rapidly developing countries such as Vietnam. The authors' objective was to demonstrate the utility of detailed injury narratives, derived from a household survey, in providing information on the determinants of work-related injuries to inform potential intervention targets. In a cross-sectional survey administered to 2615 households of a rapidly developing community of Vietnam where many workers engage in both agriculture and industrial work, the authors collected information about self-reported work-related injuries, annual hours worked in each industry and narrative text describing the circumstances of each injury. The authors used a customised coding taxonomy to describe injury scenarios. Several intervention themes emerged, including the implementation of machine guarding, the use of cut resistant gloves and safety glasses which would benefit the small- and medium-sized enterprises. Calculation of incidence rates using full-time equivalents, stratified by work group, provided some unexpected observations of the risks of working in agriculture; workers who work in agriculture in addition to another industry are at an increased risk of fatigue or overexertion and other consequences of working too hard in their agricultural activities. A lack of aggregate injury statistics makes it difficult for the owners of small- and medium-sized enterprises to recognise a priori the most effective safety interventions. This analysis of detailed injury narratives with an appropriate taxonomic basis offers the ability to focus on the level of cause, activity and source and may inform the choice of various potential interventions at the workplace or enterprise level.

Establishment of a Polychlorinated Biphenyl-Dechlorinating Microbial Consortium, Specific for Doubly Flanked Chlorines, in a Defined, Sediment-Free Medium

PubMed Central

Wu, Qingzhong; Sowers, Kevin R.; May, Harold D.

2000-01-01

Estuarine sediment from Charleston Harbor, South Carolina, was used as inoculum for the development of an anaerobic enrichment culture that specifically dechlorinates doubly flanked chlorines (i.e., chlorines bound to carbon that are flanked on both sides by other chlorine-carbon bonds) of polychlorinated biphenyls (PCBs). Dechlorination was restricted to the para chlorine in cultures enriched with 10 mM fumarate, 50 ppm (173 μM) 2,3,4,5-tetrachlorobiphenyl, and no sediment. Initially the rate of dechlorination decreased upon the removal of sediment from the medium. However, the dechlorinating activity was sustainable, and following sequential transfer in a defined, sediment-free estuarine medium, the activity increased to levels near that observed with sediment. The culture was nonmethanogenic, and molybdate, ampicillin, chloramphenicol, neomycin, and streptomycin inhibited dechlorination activity; bromoethanesulfonate and vancomycin did not. Addition of 17 PCB congeners indicated that the culture specifically removes double flanked chlorines, preferably in the para position, and does not attack ortho chlorines. This is the first microbial consortium shown to para or meta dechlorinate a PCB congener in a defined sediment-free medium. It is the second PCB-dechlorinating enrichment culture to be sustained in the absence of sediment, but its dechlorinating capabilities are entirely different from those of the other sediment-free PCB-dechlorinating culture, an ortho-dechlorinating consortium, and do not match any previously published Aroclor-dechlorinating patterns. PMID:10618202

Puerarin reduces apoptosis in rat hippocampal neurons culturea in high glucose medium by modulating the p38 mitogen activated protein kinase and c-Jun N-terminal kinase signaling pathways.

PubMed

Xu, Xiaohan; Wang, Jingbo; Zhang, Hong; Tian, Guoqing; Liu, Yuqin

2016-02-01

To investigate the neuroprotective etfect of puerarin on rat hippocampal neurons cultured in high glucose medium, and to examine the role of the p38 mitogen activated protein kinase (p38 MAPK) and c-Jun N-terminal kinase (JNK) signaling pathways in this effect. Primary cultures of hippocampal neurons were prepared from newborn Sprague Dawley rats. Neuron-specific enolase immunocytochemistry was used to identify neurons. The neurons were cultured with normal medium (control group) or with high-glucose medium (high-glucose group), and puerarin (puerarin group), a p38 MAPK inhibitor (SB239063; p38 MAPK inhibitor group) or a JNK inhibitor (SP600125; JNK inhibitor group) were added. After 72 h of treatment, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay was performed to detect apoptosis, and western blotting was used to assess protein levels of p-p38, p38, p-JNK and JNK. In the high-glucose group, the neuronal apoptosis rate and the p-p38/p38 and p-JNK/JNK ratios were higher than in the control group. The p38 MAPK and JNK inhibitors prevented this increase in the apoptosis rate. The apoptosis rates in the puerarin group, the p38 MAPK inhibitor group and the JNK inhibitor group were significantly decreased compared with the high-glucose group. Moreover, protein levels of p-p38 and p-JNK were significantly reduced, and the p-p38/p38 and p-JNK/JNK ratios were decreased in the puerarin group compared with the high-glucose group. In addition, compared with the high-glucose group, p-p38 levels and the p-p38/p38 ratio were reduced in the p38 MAPK inhibitor group, and p-JNK levels and the p-JNK/JNK ratio were decreased in the JNK inhibitor group. Puerarin attenuates neuronal apoptosis induced by high glucose by reducing the phosphorylation of p38 and JNK.

Downregulation of Lysyl Oxidase Protects Retinal Endothelial Cells From High Glucose-Induced Apoptosis.

PubMed

Kim, Dongjoon; Mecham, Robert P; Trackman, Philip C; Roy, Sayon

2017-05-01

To investigate the effect of reducing high glucose (HG)-induced lysyl oxidase (LOX) overexpression and increased activity on retinal endothelial cell apoptosis. Rat retinal endothelial cells (RRECs) were grown in normal (N) or HG (30 mM glucose) medium for 7 days. In parallel, RRECs were grown in HG medium and transfected with LOX small interfering RNA (siRNA), scrambled siRNA as control, or exposed to β-aminopropionitrile (BAPN), a LOX inhibitor. LOX expression, AKT activation, and caspase-3 activity were determined by Western blot (WB) analysis and apoptosis by differential dye staining assay. Moreover, to determine whether diabetes-induced LOX overexpression alters AKT activation and promotes apoptosis, changes in LOX expression, AKT phosphorylation, caspase-3 activation, and Bax expression were assessed in retinas of streptozotocin (STZ)-induced diabetic mice and LOX heterozygous knockout (LOX+/-) mice. WB analysis indicated significant LOX overexpression and reduced AKT activation under HG condition in RRECs. Interestingly, when cells grown in HG were transfected with LOX siRNA or exposed to BAPN, the number of apoptotic cells was significantly decreased concomitant with increased AKT phosphorylation. Diabetic mouse retinas exhibited LOX overexpression, decreased AKT phosphorylation, and increased Bax and caspase-3 activation compared to values in nondiabetic mice. In LOX+/- mice, reduced LOX levels were observed with increased AKT activity, and reduced Bax and caspase-3 activity. Furthermore, decreased levels of LOX in the LOX+/- mice was protective against diabetes-induced apoptosis. Findings from this study indicate that preventing LOX overexpression may be protective against HG-induced apoptosis in retinal vascular cells associated with diabetic retinopathy.

Somatic embryogenesis of carrot in hormone-free medium: external pH control over morphogenesis

NASA Technical Reports Server (NTRS)

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

1990-01-01

Cultures of preglobular stage proembryos (PGSPs) were initiated from mechanically wounded mature zygotic embryos of carrot, Daucus carota, on a hormone-free, semisolid medium. These PGSPs have been maintained and multiplied for extended periods without their progression into later embryo stages on the same hormone-free medium containing 1 mM NH4+ as the sole nitrogen source. Sustained maintenance of cultures comprised exclusively of PGSPs was dependent on medium pH throughout the culture period. Best growth and multiplication of PGSP cultures occurred when the pH of unbuffered, hormone-free medium fell from 4.5 to 4 over a 2-week period or when buffered medium was titrated to pH 4. If the hormone-free medium was buffered to sustain a pH at or above 4.5, PGSPs developed into later embryo stages. Maintenance with continuous multiplication of PGSPs occurred equally well on medium containing NH4+ or NH4+ and NO3-, but growth was poor with NO3- alone. Additional observations on the effects of medium components such as various nitrogen sources and levels, sucrose concentration, semisolid supports, type of buffer, borate concentration, activated charcoal, and initial pH that permit optimum maintenance of the PGSPs or foster their continued developmental progression into mature embryos and plantlets are reported. The influence of the pH of the hormone-free medium as a determinant in maintaining cultures as PGSPs or allowing their continued embryonic development are unequivocally demonstrated by gross morphology, scanning electron microscopy, and histological preparations.

[Motivation effect on EEG spectral power and heart rate parameters in students during examination stress].

PubMed

Dzhebrailova, T D; Korobeĭnikova, I I; Rudneva, L P

2014-09-01

EEG spectral power was calculated in 24 students (18-21 years) with different levels of motivation and anxiety (tested by Spielberger) in two experimental conditions: during the common educational process and the examination stress. Before examination tests, in subjects with high motivation and anxiety level the relative delta activity power increased in right frontal (F4) brain areas. In students with medium motivation immediately before an examination the relative beta2-activity power increased in right frontal (F4) brain areas. It is suggested that delta oscillati- ons reflect activity of the defensive motivational system, whereas beta2 oscillations may be associated with the achievement motivation.

Evaluation research of small and medium-sized enterprise informatization on big data

NASA Astrophysics Data System (ADS)

Yang, Na

2017-09-01

Under the background of big data, key construction of small and medium-sized enterprise informationization level was needed, but information construction cost was large, while information cost of inputs can bring benefit to small and medium-sized enterprises. This paper established small and medium-sized enterprise informatization evaluation system from hardware and software security level, information organization level, information technology application and the profit level, and information ability level. The rough set theory was used to brief indexes, and then carry out evaluation by support vector machine (SVM) model. At last, examples were used to verify the theory in order to prove the effectiveness of the method.

An ecological model of the Northern and Central Adriatic Sea: Analysis of ecosystem structure and fishing impacts

NASA Astrophysics Data System (ADS)

Coll, Marta; Santojanni, Alberto; Palomera, Isabel; Tudela, Sergi; Arneri, Enrico

2007-08-01

A trophic mass-balance model was developed to characterise the food web structure and functioning of the Northern and Central Adriatic Sea and to quantify the ecosystem impacts of fishing during the 1990s. Forty functional groups were described, including target and non-target fish and invertebrate groups, and three detritus groups (natural detritus, discards and by-catch of cetaceans and marine turtles). Results highlighted that there was an important coupling between pelagic-benthic production of plankton, benthic invertebrates and detritus. Organisms located at low and medium trophic levels, (i.e. benthic invertebrates, zooplankton and anchovy), as well as dolphins, were identified as keystone groups of the ecosystem. Jellyfish were an important element in terms of consumption and production of trophic flows within the ecosystem. The analysis of trophic flows of zooplankton and detritus groups indirectly underlined the importance of the microbial food web in the Adriatic Sea. Fishing activities inflicted notable impacts on the ecosystem during the 1990s, with a high gross efficiency of the fishery, a high consumption of fishable production, high exploitation rates for various target and non target species, a low trophic level of the catch and medium values of primary production required to sustain the fishery. Moreover, the analysis of Odum's ecological indicators highlighted that the ecosystem was in a low-medium developmental stage. Bottom trawling ( Strascico), mid-water trawling ( Volante) and beam trawling ( Rapido) fleets had the highest impacts on both target and non target ecological groups. On the contrary, purse seining ( Lampara) showed medium to low impacts on the ecosystem; cetaceans, marine turtles and sea birds were not significantly involved in competition with fishing activity.

Impact of simulated microgravity on the secretory and adhesive activity of cultured human vascular endothelial cells.

NASA Astrophysics Data System (ADS)

Rudimov, Evgeny; Buravkova, Ludmila; Pogodina, Margarita; Andrianova, Irina

The layer of vascular endothelial cells (ECs) is a dynamic,disseminated organ that perform the function of an interface between the blood and vascular wall. The endothelial monolayer is able to quickly respond to changes in the microenvironment due to its synthesis of vasoactive substances, chemokines, adhesion molecules expression, etc. ECs are highly sensitive to gravitational changes and capable of short-term and long-term responses (Sangha et al., 2001; Buravkova et al., 2005; Infanger et al., 2006, 2007. However, the question remains how to reflect the impact of microgravity on endothelium under the inflammatory process. Therefore, the aim of this study was to investigate secretory and adhesive activity of human umbilical vein endothelial cells (HUVECs) during simulated microgravity and TNF-a activation. HUVECs were isolated according to Gimbrone et al. (1978) in modification A. Antonov (1981) and used for experiments at 2-4 passages. HUVECs were activated by low level of TNF-a (2 ng/ml). Microgravity was generated by Random Positioning Machine (RPM, Dutch Space, Leiden) placed into the thermostat at 37°C. After 24 hours of clinorotation we measured adhesion molecules expression on the cell surface (ICAM-1, VCAM-1, PECAM-1, E-selectin, CD144, endoglin (CD105)) and cell viability using a flow cytometry. To evaluate the level of target gene expression was used the real time RT-PCR. IL-6 and IL-8 concentration was measured in the conditioned medium of HUVECs by using the ELISA test. We found that simulated microgravity within 24 hours caused a decrease of ICAM-1, CD144, and E-selectin expression, at the same time not affect the cell viability, endoglin and PECAM-1 expression on the surface HUVEC. Furthermore, there were no changes of the level of IL-6 and IL-8 gene expression and their products in the culture medium. TNF-activated HUVECs showed an increase in gene expression of interleukins and molecules involved in the adhesion process, which also was confirmed by the higher level of cytokines in the medium and elevated share of CD144, ICAM-1 and VCAM-1-positive cells. Comparative analysis of the level TNF-induced secretion of IL-6 and IL-8, as well as the share of cells bearing ICAM-1 and VCAM-1, showed significant variability depending on the donors. Simultaneous exposure to simulated microgravity and proinflammatory activation did not potentiate and did not cancel the effect caused by TNF-a. In summary, our findings indicate that the simulated microgravity is not activating and additional pro-inflammatory stimulus to HUVEC in vitro model. This work was supported in part by Grant from RFBR No.12-04-31763 and Grant No.NSh-371.2014.4

New medium for detection of esterase and gelatinase activity.

PubMed

Pácová, Z; Kocur, M

1984-10-01

A new medium was developed for detecting esterase and gelatinase activities in aerobic and facultatively anaerobic bacteria. The new medium was tested with various strains of bacteria and the results showed agreement between the reactions in the new medium and those obtained by conventional techniques. The new medium is more economical and may be used for a rapid differentiation of Serratia, Aeromonas and Vibrio species from biochemically similar bacteria.

Contribution of Auditory Learning Style to Students’ Mathematical Connection Ability

NASA Astrophysics Data System (ADS)

Karlimah; Risfiani, F.

2017-09-01

This paper presents the results of the research on the relation of mathematical concept with mathematics, other subjects, and with everyday life. This research reveals study result of the students who had auditory learning style and correlates it with their ability of mathematical connection. In this research, the researchers used a combination model or sequential exploratory design method, which is the use of qualitative and quantitative research methods in sequence. The result proves that giving learning facilities which are not suitable for the class whose students have the auditory learning style results in the barely sufficient math connection ability. The average mathematical connection ability of the auditory students was initially in the medium level of qualification. Then, the improvement in the form of the varied learning that suited the auditory learning style still showed the average ability of mathematical connection in medium level of qualification. Nevertheless, there was increase in the frequency of students in the medium level of qualification and decrease in the very low and low level of qualification. This suggests that the learning facilities, which are appropriate for the student’s auditory learning style, contribute well enough to the students’ mathematical connection ability. Therefore, the mathematics learning for students who have an auditory learning style should consist of particular activity that is understanding the concepts of mathematics and their relations.

Let’s Move for Pacific Islander Communities: An Evidence-Based Intervention to Increase Physical Activity

PubMed Central

LaBreche, Mandy; Cheri, Ashley; Custodio, Harold; Fex, Cleo Carlos; Foo, Mary Anne; Lepule, Jonathan Tana; May, Vanessa Tui’one; Orne, Annette; Pang, Jane Ka’ala; Pang, Victor Kaiwi; Sablan-Santos, Lola; Schmidt-Vaivao, Dorothy; Surani, Zul; Talavou, Melevesi Fifita; Toilolo, Tupou; Palmer, Paula Healani; Tanjasiri, Sora Park

2015-01-01

Pacific Islander (PI) populations of Southern California experience high obesity and low physical activity levels. Given PI’s rich cultural ties, efforts to increase physical activity using a community tailored strategy may motivate members in a more sustainable manner. In this paper, we: 1) detail the program adaptation methodology that was utilized to develop the Weaving an Islander Network for Cancer Awareness, Research and Training (WINCART) Center’s PI Let’s Move Program, a culturally-tailored program aimed to increase physical activity levels among members of PI organizations in Southern California, and 2) share the program’s pilot evaluation results on individual and organizational changes. The WINCART Center applied the National Cancer Institute’s program adaptation guidelines to tailor the evidence-based Instant Recess program to fit the needs of PIs. The end product, the PI Let’s Move Program, was piloted in 2012 with eight PI organizations, reaching 106 PI adults. At baseline, 52% of participants reported that they were not physically active, with the average number of days engaged in medium-intensity physical activity at 2.09 days/week. After the 2-month program, participants increased the number of days that they engaged in medium-intensity physical activity from 2.09 to 2.90 days/week. Post-pilot results found that 82% of participants reported intentions to engage in physical activity for at least the next six months. At baseline, only one organization was currently implementing a physical activity program, and none had implemented an evidence-based physical activity program tailored for PIs. After the 2-month timeframe, despite varying levels of capacity, all eight organizations were able to successfully implement the program. In conclusion, results from our program provide evidence that disparity populations, such as PIs, can be successfully reached through programs that are culturally tailored to both individuals and their community organizations. PMID:26153489

Metabolic Modulation by Medium-Chain Triglycerides Reduces Oxidative Stress and Ameliorates CD36-Mediated Cardiac Remodeling in Spontaneously Hypertensive Rat in the Initial and Established Stages of Hypertrophy.

PubMed

Saifudeen, Ismael; Subhadra, Lakshmi; Konnottil, Remani; Nair, R Renuka

2017-03-01

Left ventricular hypertrophy (LVH) is characterized by a decrease in oxidation of long-chain fatty acids, possibly mediated by reduced expression of the cell-surface protein cluster of differentiation 36 (CD36). Spontaneously hypertensive rats (SHRs) were therefore supplemented with medium-chain triglycerides (MCT), a substrate that bypasses CD36, based on the assumption that the metabolic modulation will ameliorate ventricular remodeling. The diet of 2-month-old and 6-month-old SHRs was supplemented with 5% MCT (Tricaprylin), for 4 months. Metabolic modulation was assessed by mRNA expression of peroxisome proliferator-activated receptor α and medium-chain acyl-CoA dehydrogenase. Blood pressure was measured noninvasively. LVH was assessed with the use of hypertrophy index, cardiomyocyte cross-sectional area, mRNA expression of B-type natriuretic peptide, cardiac fibrosis, and calcineurin-A levels. Oxidative stress indicators (cardiac malondialdehyde, protein carbonyl, and 3-nitrotyrosine levels), myocardial energy level (ATP, phosphocreatine), and lipid profile were determined. Supplementation of MCT stimulated fatty acid oxidation in animals of both age groups, reduced hypertrophy and oxidative stress along with the maintenance of energy level. Blood pressure, body weight, and lipid profile were unaffected by the treatment. The results indicate that modulation of myocardial fatty acid metabolism by MCT prevents progressive cardiac remodeling in SHRs, possibly by maintenance of energy level and decrease in oxidative stress. Copyright © 2016 Elsevier Inc. All rights reserved.

Evaluation and optimization of hepatocyte culture media factors by design of experiments (DoE) methodology

PubMed Central

Dong, Jia; Lübberstedt, Marc; Urbaniak, Thomas; Nüssler, Andreas K.N.; Knobeloch, Daniel; Gerlach, Jörg C.; Zeilinger, Katrin

2008-01-01

Optimization of cell culture media based on statistical experimental design methodology is a widely used approach for improving cultivation conditions. We applied this methodology to refine the composition of an established culture medium for growth of a human hepatoma cell line, C3A. A selection of growth factors and nutrient supplements were systematically screened according to standard design of experiments (DoE) procedures. The results of the screening indicated that the medium additives hepatocyte growth factor, oncostatin M, and fibroblast growth factor 4 significantly influenced the metabolic activities of the C3A cell line. Surface response methodology revealed that the optimum levels for these factors were 30 ng/ml for hepatocyte growth factor and 35 ng/ml for oncostatin M. Additional experiments on primary human hepatocyte cultures showed high variance in metabolic activities between cells from different individuals, making determination of optimal levels of factors more difficult. Still, it was possible to conclude that hepatocyte growth factor, epidermal growth factor, and oncostatin M had decisive effects on the metabolic functions of primary human hepatocytes. PMID:19003182

Evaluation and optimization of hepatocyte culture media factors by design of experiments (DoE) methodology.

PubMed

Dong, Jia; Mandenius, Carl-Fredrik; Lübberstedt, Marc; Urbaniak, Thomas; Nüssler, Andreas K N; Knobeloch, Daniel; Gerlach, Jörg C; Zeilinger, Katrin

2008-07-01

Optimization of cell culture media based on statistical experimental design methodology is a widely used approach for improving cultivation conditions. We applied this methodology to refine the composition of an established culture medium for growth of a human hepatoma cell line, C3A. A selection of growth factors and nutrient supplements were systematically screened according to standard design of experiments (DoE) procedures. The results of the screening indicated that the medium additives hepatocyte growth factor, oncostatin M, and fibroblast growth factor 4 significantly influenced the metabolic activities of the C3A cell line. Surface response methodology revealed that the optimum levels for these factors were 30 ng/ml for hepatocyte growth factor and 35 ng/ml for oncostatin M. Additional experiments on primary human hepatocyte cultures showed high variance in metabolic activities between cells from different individuals, making determination of optimal levels of factors more difficult. Still, it was possible to conclude that hepatocyte growth factor, epidermal growth factor, and oncostatin M had decisive effects on the metabolic functions of primary human hepatocytes.

Thin disk laser with unstable resonator and reduced output coupler

NASA Astrophysics Data System (ADS)

Gavili, Anwar; Shayganmanesh, Mahdi

2018-05-01

In this paper, feasibility of using unstable resonator with reduced output coupling in a thin disk laser is studied theoretically. Unstable resonator is modeled by wave-optics using Collins integral and iterative method. An Yb:YAG crystal with 250 micron thickness is considered as a quasi-three level active medium and modeled by solving rate equations of energy levels populations. The amplification of laser beam in the active medium is calculated based on the Beer-Lambert law and Rigrod method. Using generalized beam parameters method, laser beam parameters like, width, divergence, M2 factor, output power as well as near and far-field beam profiles are calculated for unstable resonator. It is demonstrated that for thin disk laser (with single disk) in spite of the low thickness of the disk which leads to low gain factor, it is possible to use unstable resonator (with reduced output coupling) and achieve good output power with appropriate beam quality. Also, the behavior of output power and beam quality versus equivalent Fresnel number is investigated and optimized value of output coupling for maximum output power is achieved.

The galactolipase activity of Fusarium solani (phospho)lipase.

PubMed

Jallouli, Raida; Othman, Houcemeddine; Amara, Sawsan; Parsiegla, Goetz; Carriere, Frédéric; Srairi-Abid, Najet; Gargouri, Youssef; Bezzine, Sofiane

2015-03-01

The purified (phospho)lipase of Fusarium solani (FSL), was known to be active on both triglycerides and phospholipids. This study aimed at assessing the potential of this enzyme in hydrolyzing galactolipids. FSL was found to hydrolyze at high rates of synthetic medium chains monogalactosyldiacylglycerol (4658±146U/mg on DiC8-MGDG) and digalactosyldiacylglycerol (3785±83U/mg on DiC8-DGDG) and natural long chain monogalactosyldiacylglycerol extracted from leek leaves (991±85U/mg). It is the microbial enzyme with the highest activity on galactolipids identified so far with a level of activity comparable to that of pancreatic lipase-related protein 2. FSL maximum activity on galactolipids was measured at pH8. The analysis of the hydrolysis product of natural MGDG from leek showed that FSL hydrolyzes preferentially the ester bond at the sn-1 position of galactolipids. To investigate the structure-activity relationships of FSL, a 3D model of this enzyme was built. In silico docking of medium chains MGDG and DGDG and phospholipid in the active site of FSL reveals structural solutions which are in concordance with in vitro tests. Copyright © 2015 Elsevier B.V. All rights reserved.

Regulation of phosphoenolpyruvate carboxykinase and pyruvate kinase in Saccharomyces cerevisiae grown in the presence of glycolytic and gluconeogenic carbon sources and the role of mitochondrial function on gluconeogenesis.

PubMed

Wilson, A J; Bhattacharjee, J K

1986-12-01

Phosphoenolpyruvate carboxykinase (PEPCKase) and pyruvate kinase (PKase) were measured in Saccharomyces cerevisiae grown in the presence of glycolytic and gluconeogenic carbon sources. The PEPCKase activity was highest in ethanol-grown cells. However, high PEPCKase activity was also observed in cells grown in 1% glucose, especially as compared with the activity of sucrose-, maltose-, or galactose-grown cells. Activity was first detected after 12 h when glucose was exhausted from the growth medium. The PKase activity was very high in glucose-grown cells; considerable activity was also present in ethanol- and pyruvate-grown cells. The absolute requirement of respiration for gluconeogenesis was demonstrated by the absence or significantly low levels of PEPCKase and fructose-1,6-bisphosphatase activities observed in respiratory deficient mutants, as well as in wild-type S. cerevisiae cells grown in the presence of glucose and antimycin A or chloramphenicol. Obligate glycolytic and gluconeogenic enzymes were present simultaneously only in stationary phase cells, but not in exponential phase cells; hence futile cycling could not occur in log phase cells regardless of the presence of carbon source in the growth medium.

Neuronal medium that supports basic synaptic functions and activity of human neurons in vitro.

PubMed

Bardy, Cedric; van den Hurk, Mark; Eames, Tameji; Marchand, Cynthia; Hernandez, Ruben V; Kellogg, Mariko; Gorris, Mark; Galet, Ben; Palomares, Vanessa; Brown, Joshua; Bang, Anne G; Mertens, Jerome; Böhnke, Lena; Boyer, Leah; Simon, Suzanne; Gage, Fred H

2015-05-19

Human cell reprogramming technologies offer access to live human neurons from patients and provide a new alternative for modeling neurological disorders in vitro. Neural electrical activity is the essence of nervous system function in vivo. Therefore, we examined neuronal activity in media widely used to culture neurons. We found that classic basal media, as well as serum, impair action potential generation and synaptic communication. To overcome this problem, we designed a new neuronal medium (BrainPhys basal + serum-free supplements) in which we adjusted the concentrations of inorganic salts, neuroactive amino acids, and energetic substrates. We then tested that this medium adequately supports neuronal activity and survival of human neurons in culture. Long-term exposure to this physiological medium also improved the proportion of neurons that were synaptically active. The medium was designed to culture human neurons but also proved adequate for rodent neurons. The improvement in BrainPhys basal medium to support neurophysiological activity is an important step toward reducing the gap between brain physiological conditions in vivo and neuronal models in vitro.

Neuronal medium that supports basic synaptic functions and activity of human neurons in vitro

PubMed Central

Bardy, Cedric; van den Hurk, Mark; Eames, Tameji; Marchand, Cynthia; Hernandez, Ruben V.; Kellogg, Mariko; Gorris, Mark; Galet, Ben; Palomares, Vanessa; Brown, Joshua; Bang, Anne G.; Mertens, Jerome; Böhnke, Lena; Boyer, Leah; Simon, Suzanne; Gage, Fred H.

2015-01-01

Human cell reprogramming technologies offer access to live human neurons from patients and provide a new alternative for modeling neurological disorders in vitro. Neural electrical activity is the essence of nervous system function in vivo. Therefore, we examined neuronal activity in media widely used to culture neurons. We found that classic basal media, as well as serum, impair action potential generation and synaptic communication. To overcome this problem, we designed a new neuronal medium (BrainPhys basal + serum-free supplements) in which we adjusted the concentrations of inorganic salts, neuroactive amino acids, and energetic substrates. We then tested that this medium adequately supports neuronal activity and survival of human neurons in culture. Long-term exposure to this physiological medium also improved the proportion of neurons that were synaptically active. The medium was designed to culture human neurons but also proved adequate for rodent neurons. The improvement in BrainPhys basal medium to support neurophysiological activity is an important step toward reducing the gap between brain physiological conditions in vivo and neuronal models in vitro. PMID:25870293

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dewitt, Ann E.; Dong, Jian Y.; Wiley, H S.

Autocrine signaling is important in normal tissue physiology as well as pathological conditions. It is difficult to analyze these systems, however, because they are both self-contained and recursive. To understand how parameters, such as ligand production and receptor expression influence autocrine activity, we investigated a human epidermal growth factor/epidermal growth factor receptor (EGF/EGFR) loop engineered into mouse B82 fibroblasts. We varied the level of ligand production using the tet-off expression system and used metalloprotease inhibitors to modulate ligand release. Receptor expression was varied using antagonistic, blocking antibodies. We compared autocrine ligand release to receptor activation using a microphysiometer-based assay andmore » analyzed our data with a quantitative model of ligand release and receptor dynamics. We found that the activity of our autocrine system could be described in terms of a simple ratio between the rate of ligand production (VL) and the rate of receptor production (VR). At a VL/VR ratio of < 0.3, essentially no ligand was found in the extracellular medium, but a significant number cell receptors (30-40%) were occupied. As the VL/VR ratio increased from 0.3 towards unity, receptor occupancy increased, and significant amounts of ligand now appeared in the medium. Above a VL/VR ratio of 1.0, receptor occupancy approached saturation and most of the released ligand was lost into the medium. Analysis of human mammary epithelial cells showed that a VL/VR ratio of < 5 x 10 -4 was sufficient to evoke >20% of a maximal proliferative response. This suggests that natural autocrine systems are active even when no ligand appears in the extracellular medium; i.e., they operate 'invisibly' to general detection.« less

Lactic acid production from submerged fermentation of broken rice using undefined mixed culture.

PubMed

Nunes, Luiza Varela; de Barros Correa, Fabiane Fernanda; de Oliva Neto, Pedro; Mayer, Cassia Roberta Malacrida; Escaramboni, Bruna; Campioni, Tania Sila; de Barros, Natan Roberto; Herculano, Rondinelli Donizetti; Fernández Núñez, Eutimio Gustavo

2017-04-01

The present work aimed to characterize and optimize the submerged fermentation of broken rice for lactic acid (LA) production using undefined mixed culture from dewatered activated sludge. A microorganism with amylolytic activity, which also produces LA, Lactobacillus amylovorus, was used as a control to assess the extent of mixed culture on LA yield. Three level full factorial designs were performed to optimize and define the influence of fermentation temperature (20-50 °C), gelatinization time (30-60 min) and broken rice concentration in culture medium (40-80 g L -1 ) on LA production in pure and undefined mixed culture. LA production in mixed culture (9.76 g L -1 ) increased in sixfold respect to pure culture in optimal assessed experimental conditions. The optimal conditions for maximizing LA yield in mixed culture bioprocess were 31 °C temperature, 45 min gelatinization time and 79 g L -1 broken rice concentration in culture medium. This study demonstrated the positive effect of undefined mixed culture from dewatered activated sludge to produce LA from culture medium formulated with broken rice. In addition, this work establishes the basis for an efficient and low-cost bioprocess to manufacture LA from this booming agro-industrial by-product.

Hexose Transport in Growing Petunia Pollen Tubes and Characterization of a Pollen-Specific, Putative Monosaccharide Transporter1

PubMed Central

Ylstra, Bauke; Garrido, Dolores; Busscher, Jacqueline; van Tunen, Arjen J.

1998-01-01

We investigated the molecular and physiological processes of sugar uptake and metabolism during pollen tube growth and plant fertilization. In vitro germination assays showed that petunia (Petunia hybrida) pollen can germinate and grow not only in medium containing sucrose (Suc) as a carbon source, but also in medium containing the monosaccharides glucose (Glc) or fructose (Fru). Furthermore, high-performance liquid chromatography analysis demonstrated a rapid and complete conversion of Suc into equimolar amounts of Glc and Fru when pollen was cultured in a medium containing 2% Suc. This indicates the presence of wall-bound invertase activity and uptake of sugars in the form of monosaccharides by the growing pollen tube. A cDNA designated pmt1 (petunia monosaccharide transporter 1), which is highly homologous to plant monosaccharide transporters, was isolated from petunia. Pmt1 belongs to a small gene family and is expressed specifically in the male gametophyte, but not in any other vegetative or floral tissues. Pmt1 is activated after the first pollen mitosis, and high levels of mRNA accumulate in mature and germinating pollen. A model describing the transport of sugars to the style, the conversion of Suc into Glc and Fru, and the active uptake by a monosaccharide transporter into the pollen tube is presented. PMID:9733549

[Optimization of cultural condition of genetic engineering strain for antibiotic peptide adenoregulin and research on its fed-batch cultivation].

PubMed

Zhou, Yu-Xun; Cao, Wei; Wei, Dong-Zhi; Luo, Qing-Ping; Wang, Jin-Zhi

2005-07-01

33 amino acid antibiotic peptide adenoregulin (ADR), which were firstly isolated from the skin of South America arboreal frog Phyllomedusa bicolor, forms alpha-helix amphipathic structure in apolar medium and has a wide spectrum of antimicrobial activity and high potency of lytic ability. Adr gene was cloned in pET32a and transformed into Escherichia coli BL21(DE3) . The cultural and inductive conditions of E. coli BL21(DE3)/pET32a-adr have been optimized. The effect of three factors which were time point of induction, concentration of IPTG in the culture and time of induction on the expression level of Trx-ADR was investigated. The results indicated that the expression level was affected by the time point of induction most predominantly. 9 veriaties of media in which BL21 (DE3)/pET32a-adr was cultured and induced were tested to achieve high expression level of target protein. It was found that glucose in the medium played an important role in keeping stable and high expression level of Trx-ADR. The optimal inductive condition is as follows: the culture medium is 2 x YT + 0.5% glucose, the time point of induction is OD600 = 0.9, the final concentration of IPTG in the culture is 0.1 mmol/L and the induction time is 4 h. BL21 (DE3)/pET32a-adr was cultivated according to the strategy of constant pH at early stage and exponential feeding at later stage to obtain high cell density. During the entire fed-batch phase, by controlling the feeding of glucose, the specific growth rate of the culture was controlled at about 0.15 h(-1), the accumulation of acetic acid was controlled at low level (<2 g/L), but the plasmid stability could not be maintained well. At the end of the cultivation, 40% of the bacteria in the culture lost their plasmids. As a result, the expression level of the target protein declined dramatically, but 90% of Trx-ADR was in soluble form. The expressed fusion protein showed no antibacterial activity, while the native form of ADR lysed from Trx-ADR showed distinct antibacterial activity.

Metabolic Mapping: Quantitative Enzyme Cytochemistry and Histochemistry to Determine the Activity of Dehydrogenases in Cells and Tissues.

PubMed

Molenaar, Remco J; Khurshed, Mohammed; Hira, Vashendriya V V; Van Noorden, Cornelis J F

2018-05-26

Altered cellular metabolism is a hallmark of many diseases, including cancer, cardiovascular diseases and infection. The metabolic motor units of cells are enzymes and their activity is heavily regulated at many levels, including the transcriptional, mRNA stability, translational, post-translational and functional level. This complex regulation means that conventional quantitative or imaging assays, such as quantitative mRNA experiments, Western Blots and immunohistochemistry, yield incomplete information regarding the ultimate activity of enzymes, their function and/or their subcellular localization. Quantitative enzyme cytochemistry and histochemistry (i.e., metabolic mapping) show in-depth information on in situ enzymatic activity and its kinetics, function and subcellular localization in an almost true-to-nature situation. We describe a protocol to detect the activity of dehydrogenases, which are enzymes that perform redox reactions to reduce cofactors such as NAD(P) + and FAD. Cells and tissue sections are incubated in a medium that is specific for the enzymatic activity of one dehydrogenase. Subsequently, the dehydrogenase that is the subject of investigation performs its enzymatic activity in its subcellular site. In a chemical reaction with the reaction medium, this ultimately generates blue-colored formazan at the site of the dehydrogenase's activity. The formazan's absorbance is therefore a direct measure of the dehydrogenase's activity and can be quantified using monochromatic light microscopy and image analysis. The quantitative aspect of this protocol enables researchers to draw statistical conclusions from these assays. Besides observational studies, this technique can be used for inhibition studies of specific enzymes. In this context, studies benefit from the true-to-nature advantages of metabolic mapping, giving in situ results that may be physiologically more relevant than in vitro enzyme inhibition studies. In all, metabolic mapping is an indispensable technique to study metabolism at the cellular or tissue level. The technique is easy to adopt, provides in-depth, comprehensive and integrated metabolic information and enables rapid quantitative analysis.

Long-term biases in geomagnetic K and aa indices

USGS Publications Warehouse

Love, J.J.

2011-01-01

Analysis is made of the geomagnetic-activity aa index and its source K-index data from groups of ground-based observatories in Britain, and Australia, 1868.0-2009.0, solar cycles 11-23. The K data show persistent biases, especially for high (low) K-activity levels at British (Australian) observatories. From examination of multiple subsets of the K data we infer that the biases are not predominantly the result of changes in observatory location, localized induced magnetotelluric currents, changes in magnetometer technology, or the modernization of K-value estimation methods. Instead, the biases appear to be artifacts of the latitude-dependent scaling used to assign K values to particular local levels of geomagnetic activity. The biases are not effectively removed by weighting factors used to estimate aa. We show that long-term averages of the aa index, such as annual averages, are dominated by medium-level geomagnetic activity levels having K values of 3 and 4. ?? 2011 Author(s).

Serum biomarkers for acute hepatotoxicity of Echis pyramidum snake venom in rats.

PubMed

Asmari, Abdulrahman K Al; Khan, Haseeb A; Banah, Faisal A; Buraidi, Ahmed A Al; Manthiri, Rajamohammed A

2015-01-01

Echis pyramidum is a venomous viper responsible for most cases of envenomation in Arabian Peninsula. We determined the acute phase (3-6 h) changes in serum markers of liver function including alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT) and bilirubin in adult male Sprague Dawley rats injected with Echis pyramidum venom (EPV) in the doses of 0.00 (control), 0.25, 0.50 and 1.00 mg/kg bodyweight. We also analyzed markers of oxidative stress including superoxide dismutase (SOD), catalase (CAT), total thiols (T-SH) and thiobarbituric acids reactive substances (TBARS) in liver. The results showed significant and dose- and time-dependent increases in serum ALT, ALP and GGT activities after a single injection of EPV. Serum bilirubin was significantly increased by medium and high doses of EVP after 3 h post-injection and then decreased at 6 h. The low dose of EPV neither affected the activity of SOD nor altered the levels of liver T-SH and TBARS, however, it significantly decreased the activity of CAT at 6 h post-injection of EPV. The medium dose of EPV significantly reduced liver SOD activity after 6 h whereas the high dose significantly reduced the SOD activity at 3 h and 6 h post-dosing. Both medium and high doses of EPV caused significant as well as dose- and time-dependent reductions in liver CAT activities. The high dose significantly reduced T-SH and increased TBARS in rat liver. Further studies are warranted to test the pharmacological potential of early phase antioxidant therapy for neutralizing the toxic effects of EPV.

Screening and production of a potent extracellular Arthrobacter creatinolyticus urease for determination of heavy metal ions.

PubMed

Ramesh, Rajendran; Aarthy, Mayilvahanan; Gowthaman, Marichetti Kuppuswami; Gabrovska, Katya; Godjevargova, Tzonka; Kamini, Numbi Ramudu

2014-04-01

This paper describes the isolation of a potent extracellular urease producing microorganism, identified by 16S rRNA as Arthrobacter creatinolyticus MTCC 5604 and its medium optimization by classical one-factor-at-a-time method and central composite rotatable design (CCRD), a tool of response surface methodology (RSM). An optimal activity of 9.0 U ml(-1) was obtained by classical method and statistical optimization of the medium resulted in an activity of 17.35 U ml(-1) at 48 h and 30 °C. This activity was 4.91 times greater than the initial activity (3.53 U ml(-1) ) from the basal medium and the enzyme showed maximum activity at pH 8.0 and 60 °C and was stable at pH 7.0-9.0 and temperatures up to 50 °C. Furthermore, the enzyme was assessed for its activity reduction by determining the inhibitory concentration (IC50 ) of heavy metal ions and the inhibition of urease was in the order of Cu(II) > Cd(II) > Zn(II) > Ni(II). Urease was highly sensitive to Cu(II) and its inhibition was 94% and 100% in model solutions containing a mixture of Cu(II) with heavy metal ions Cd(II) and Zn(II), respectively. The results of these studies suggested that the enzyme could be utilized as sensors to determine the levels of Cu(II) ions in industrial effluents, contaminated soil and ground water. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Prognostic Value of Soluble Death Receptor Ligands in Patients with Transitional Cell Carcinoma of Bladder.

PubMed

Ben Bahria-Sediki, Islem; Chebil, Mohamed; Sampaio, Carla; Martel-Frachet, Véronique; Cherif, Mohamed; Zermani, Rachida; Rammeh, Soumaya; Ben Ammar Gaaied, Amel; Bettaieb, Ali

2018-05-02

The activation of Fas/Fas ligand (FasL) and DR4-DR5/tumor necrosis factor-related-apoptosis-inducing ligand (TRAIL) pathways in cancer cells triggers apoptosis. The objective of this study was to investigate the prognostic value of soluble FasL (sFasL) and soluble (sTRAIL) in the serum of patients with bladder cancer. The sFasL and sTRAIL levels in the sera of patients with bladder cancer or healthy donors were determined using the enzyme-linked immunosorbent assay. Micro-culture tetrazolium viability assay and Western blot were used to analyze cell cytotoxicity and death receptors protein expression respectively. Whether no difference in sTRAIL levels was seen between patients and controls, the level of sFasL was higher in patients than that in healthy donors. According to, sFasL level was the highest in the serum of patients with superficial stage or low- and medium-grade cancer. Moreover, sFasL in patients with superficial noninvasive bladder tumors or low- and medium-grade cancers was higher than that in patients with invasive carcinomas and high-grade cancers. Patients with high levels of sFasL survive longer than those with low levels, probably related to the cytotoxic potential of FasL preserved in its soluble form. The data suggest that monitoring the level of sFasL and its cytotoxic activity could be a prognostic marker in the follow-up of patients with bladder cancer. © 2018 S. Karger AG, Basel.

Establishment of an efficient in vitro regeneration protocol for rapid and mass propagation of Dendrobium chrysotoxum Lindl. using seed culture.

PubMed

Nongdam, Potshangbam; Tikendra, Leimapokpam

2014-01-01

An efficient in vitro regeneration protocol from seed culture has been established successfully for Dendrobium chrysotoxum, an epiphytic orchid having tremendous ornamental and medicinal values. Seed germination response was encouraging in Mitra (M) medium enriched with different combinations of auxins and cytokinins. Medium supplemented with 0.4% activated charcoal (AC), 2 mg/L 6-benzyl amino purine (BAP), and 2 mg/L indole-3-acetic acid (IAA) produced best seed germination percentage in 2 weeks of culture. Incorporation of higher concentration of kinetin (KN) or BAP in combination with low auxin in medium induced pronounced shooting and leaf formation. Reduction in leaf development was evident when cytokinins exist singly in medium indicating synergistic effect of auxin and cytokinin in leaf induction. Presence of elevated level of indole-3-butyric acid (IBA) or 1-naphthalene acetic acid (NAA) with low cytokinin content in medium generated more in vitro rooting, though IBA was found to be more effective in rooting induction as compared to NAA. The in vitro protocol for asymbiotic seed germination developed from the present investigation can be used for rapid mass propagation of this highly important Dendrobium orchid species.

Establishment of an Efficient In Vitro Regeneration Protocol for Rapid and Mass Propagation of Dendrobium chrysotoxum Lindl. Using Seed Culture

PubMed Central

2014-01-01

An efficient in vitro regeneration protocol from seed culture has been established successfully for Dendrobium chrysotoxum, an epiphytic orchid having tremendous ornamental and medicinal values. Seed germination response was encouraging in Mitra (M) medium enriched with different combinations of auxins and cytokinins. Medium supplemented with 0.4% activated charcoal (AC), 2 mg/L 6-benzyl amino purine (BAP), and 2 mg/L indole-3-acetic acid (IAA) produced best seed germination percentage in 2 weeks of culture. Incorporation of higher concentration of kinetin (KN) or BAP in combination with low auxin in medium induced pronounced shooting and leaf formation. Reduction in leaf development was evident when cytokinins exist singly in medium indicating synergistic effect of auxin and cytokinin in leaf induction. Presence of elevated level of indole-3-butyric acid (IBA) or 1-naphthalene acetic acid (NAA) with low cytokinin content in medium generated more in vitro rooting, though IBA was found to be more effective in rooting induction as compared to NAA. The in vitro protocol for asymbiotic seed germination developed from the present investigation can be used for rapid mass propagation of this highly important Dendrobium orchid species. PMID:25401154

Role of Protein Kinase C Epsilon in Prostate Cancer and Metastasis

DTIC Science & Technology

2013-08-01

detected by Western blot using an anti-Rac1 antibody. Depletion of PKCε with either #4 or #8 RNAi duplexed reduced Rac1 activation levels under serum ... serum deprivation condition was not abolished when PKCε was knocked down and therefore Rac1 activity was reduced. Although these results may implicate...medium supplemented with 10% serum . After different times the number of cells attached was quantified by MTT assay. (e) RWPE-1 cells were infected

Protection against salt toxicity in Azolla pinnata-Anabaena azollae symbiotic association by using combined-N sources.

PubMed

Mishra, A K; Singh, Satya S

2006-09-01

Protection from salt stress was observed in the terms of yield (fresh and dry weight, chlorophyll and protein) and nitrogenase activity. Azollapinnata appeared highly sensitive to 40 mM external NaCl stress. Fronds of Azolla unable to grow beyond a concentration of 30 mM NaCl and accordingly death was recorded at 40 mM NaCl on the 6th day of incubation. Yield was inhibited by various levels of NaCl (0, 10, 20 and 30 mM). Addition of combined-N to the growth medium protected the association partially from salt toxicity. Among the N-sources (NO3-, NH4+ and urea) tried, urea mitigated the salt-induced toxicity most efficiently. Reduction in nitrogenase activity was observed when intact Azolla was grown in nutrient medium either supplemented with different levels of NaCl or combined nitrogen. Only NO3- (5 mM) protected the enzymatic activity from salt toxicity while other concentrations of ammonium, nitrate and urea slowed down the salt-induced inhibition of enzyme activity in Azolla-Anabaena association. These results suggested that an optimum protection from salt stress could be obtained by using a combination of combined nitrogen sources. The reason for this protection might be due to the availability of combined nitrogen to the association, nitrogen is only available through the biological nitrogen fixation which is the most sensitive to salt stress.

Citric acid assisted phytoremediation of copper by Brassica napus L.

PubMed

Zaheer, Ihsan Elahi; Ali, Shafaqat; Rizwan, Muhammad; Farid, Mujahid; Shakoor, Muhammad Bilal; Gill, Rafaqa Ali; Najeeb, Ullah; Iqbal, Naeem; Ahmad, Rehan

2015-10-01

Use of organic acids for promoting heavy metals phytoextraction is gaining worldwide attention. The present study investigated the influence of citric acid (CA) in enhancing copper (Cu) uptake by Brassica napus L. seedlings. 6 Weeks old B. napus seedlings were exposed to different levels of copper (Cu, 0, 50 and 100µM) alone or with CA (2.5mM) in a nutrient medium for 40 days. Exposure to elevated Cu levels (50 and 100µM) significantly reduced the growth, biomass production, chlorophyll content, gas exchange attributes and soluble proteins of B. napus seedlings. In addition, Cu toxicity increased the production of hydrogen peroxide (H2O2), malondialdehyde (MDA) and electrolyte leakage (EL) in leaf and root tissues of B. napus. Activities of antioxidant enzymes such as guaiacol peroxidase (POD), superoxide dismutase (SOD), catalases (CAT), ascorbate peroxidase (APX) in root and shoot tissues of B. napus were increased in response to lower Cu concentration (50µM) but increased under higher Cu concentration (100µM). Addition of CA into nutrient medium significantly alleviated Cu toxicity effects on B. napus seedlings by improving photosynthetic capacity and ultimately plant growth. Increased activities of antioxidant enzymes in CA-treated plants seems to play a role in capturing of stress-induced reactive oxygen species as was evident from lower level of H2O2, MDA and EL in CA-treated plants. Increasing Cu concentration in the nutrient medium significantly increased Cu concentration in in B. napus tissues. Cu uptake was further increased by CA application. These results suggested that CA might be a useful strategy for increasing phytoextraction of Cu from contaminated soils. Copyright © 2015 Elsevier Inc. All rights reserved.

Green tea (Camellia sinensis) administration induces expression of immune relevant genes and biochemical parameters in rainbow trout (Oncorhynchus mykiss).

PubMed

Nootash, Shahab; Sheikhzadeh, Najmeh; Baradaran, Behzad; Oushani, Ali Khani; Maleki Moghadam, Mohammad Reza; Nofouzi, Katayoon; Monfaredan, Amir; Aghebati, Leili; Zare, Fatemeh; Shabanzadeh, Sadigheh

2013-12-01

Present study elucidates the efficacy of green tea (Camellia sinensis) on growth performance, immune and antioxidant systems and cytokine gene expression in rainbow trout tissues. Green tea was supplemented at 20, 100, and 500 mg kg(-1) diet and fed to fish (average weight: 23.5 g) for 35 days. No remarkable changes in growth performance were observed among all test groups. Lower lipid peroxidation product and higher superoxide dismutase activity were noted in fish received the medium dose of green tea. Significant increase in serum bactericidal activity and total protein were recorded in all treatment groups. All doses of green tea up-regulated Interleukin-1β transcription in the spleen, while Interleukin-1β mRNA level decreased significantly in the kidney of low dose of green tea. Interleukin-6 mRNA level was up-regulated in the spleen of high dose of green tea and liver of middle and high doses of green tea. High dose and medium dose of green tea up-regulated the interleukin-8 transcription in the kidney and liver, respectively. Meanwhile, green tea inhibited the production of interleukin-10 in all treatment groups compared with control group. Medium dose of green tea up-regulated tumor necrosis factor-α transcription in all fish tissues, while high dose and low dose of green tea enhanced tumor necrosis factor-α mRNA levels in the kidney and spleen, respectively. Present study suggests that green tea especially at 100 mg kg(-1) feed may effectively enhance the antioxidant system and immune system in rainbow trout. Copyright © 2013 Elsevier Ltd. All rights reserved.

Valproic acid attenuates skeletal muscle wasting by inhibiting C/EBPβ-regulated atrogin1 expression in cancer cachexia.

PubMed

Sun, Rulin; Zhang, Santao; Hu, Wenjun; Lu, Xing; Lou, Ning; Yang, Zhende; Chen, Shaoyong; Zhang, Xiaoping; Yang, Hongmei

2016-07-01

Muscle wasting is the hallmark of cancer cachexia and is associated with poor quality of life and increased mortality. Valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, has important biological effects in the treatment of muscular dystrophy. To verify whether VPA could ameliorate muscle wasting induced by cancer cachexia, we explored the role of VPA in two cancer cachectic mouse models [induced by colon-26 (C26) adenocarcinoma or Lewis lung carcinoma (LLC)] and atrophied C2C12 myotubes [induced by C26 cell conditioned medium (CCM) or LLC cell conditioned medium (LCM)]. Our data demonstrated that treatment with VPA increased the mass and cross-sectional area of skeletal muscles in tumor-bearing mice. Furthermore, treatment with VPA also increased the diameter of myotubes cultured in conditioned medium. The skeletal muscles in cachectic mice or atrophied myotubes treated with VPA exhibited reduced levels of CCAAT/enhancer binding protein beta (C/EBPβ), resulting in atrogin1 downregulation and the eventual alleviation of muscle wasting and myotube atrophy. Moreover, atrogin1 promoter activity in myotubes was stimulated by CCM via activating the C/EBPβ-responsive cis-element and subsequently inhibited by VPA. In contrast to the effect of VPA on the levels of C/EBPβ, the levels of inactivating forkhead box O3 (FoxO3a) were unaffected. In summary, VPA attenuated muscle wasting and myotube atrophy and reduced C/EBPβ binding to atrogin1 promoter locus in the myotubes. Our discoveries indicate that HDAC inhibition by VPA might be a promising new approach for the preservation of skeletal muscle in cancer cachexia. Copyright © 2016 the American Physiological Society.

High energy bursts from a solid state laser operated in the heat capacity limited regime

DOEpatents

Albrecht, G.; George, E.V.; Krupke, W.F.; Sooy, W.; Sutton, S.B.

1996-06-11

High energy bursts are produced from a solid state laser operated in a heat capacity limited regime. Instead of cooling the laser, the active medium is thermally well isolated. As a result, the active medium will heat up until it reaches some maximum acceptable temperature. The waste heat is stored in the active medium itself. Therefore, the amount of energy the laser can put out during operation is proportional to its mass, the heat capacity of the active medium, and the temperature difference over which it is being operated. The high energy burst capacity of a heat capacity operated solid state laser, together with the absence of a heavy, power consuming steady state cooling system for the active medium, will make a variety of applications possible. Alternately, cooling takes place during a separate sequence when the laser is not operating. Industrial applications include new material working processes. 5 figs.

High energy bursts from a solid state laser operated in the heat capacity limited regime

DOEpatents

Albrecht, Georg; George, E. Victor; Krupke, William F.; Sooy, Walter; Sutton, Steven B.

1996-01-01

High energy bursts are produced from a solid state laser operated in a heat capacity limited regime. Instead of cooling the laser, the active medium is thermally well isolated. As a result, the active medium will heat up until it reaches some maximum acceptable temperature. The waste heat is stored in the active medium itself. Therefore, the amount of energy the laser can put out during operation is proportional to its mass, the heat capacity of the active medium, and the temperature difference over which it is being operated. The high energy burst capacity of a heat capacity operated solid state laser, together with the absence of a heavy, power consuming steady state cooling system for the active medium, will make a variety of applications possible. Alternately, cooling takes place during a separate sequence when the laser is not operating. Industrial applications include new material working processes.

Effects of phenolic constituents of daylily flowers on corticosterone- and glutamate-treated PC12 cells.

PubMed

Tian, Huan; Yang, Fei-Fei; Liu, Chun-Yu; Liu, Xin-Min; Pan, Rui-Le; Chang, Qi; Zhang, Ze-Sheng; Liao, Yong-Hong

2017-01-21

Daylily flowers, the flower and bud parts of Hemerocallis citrina or H. fulva, are well known as Wang-You-Cao in Chinese, meaning forget-one's sadness plant. However, the major types of active constituents responsible for the neurological effects remain unclear. This study was to examine the protective effects of hydroalcoholic extract and fractions and to identify the active fractions. The extract of daylily flowers was separated with AB-8 resin into different fractions containing non-phenolic compounds, phenolic acid derivatives and flavonoids as determined using UPLC-DAD chromatograms. The neuroprotective activity was measured by evaluating the cell viability and lactate dehydrogenase release using PC12 cell damage models induced by corticosterone and glutamate. The neurological mechanisms were explored by determining their effect on the levels of dopamine (DA), 5-hydroxy tryptamine (5-HT), γ-aminobutyric acid (GABA), noradrenaline (NE) and acetylcholine (ACh) in the cell culture medium measured using an LC-MS/MS method. Pretreatment of PC12 cells with the extract and phenolic fractions of daylily flowers at concentrations ranging from 0.63 to 5 mg raw material/mL significantly reversed corticosterone- and glutamate-induced neurotoxicity in a dose-dependent manner. The fractions containing phenolic acid derivatives (0.59% w/w in the flowers) and/or flavonoids (0.60% w/w) exerted similar dose-dependent neuroprotective effect whereas the fractions with non-phenolic compounds exhibited no activity. The presence of phenolic acid derivatives in the corticosterone- and glutamate-treated PC12 cells elevated the DA level in the cell culture medium whereas flavonoids resulted in increased ACH and 5-HT levels. Phenolic acid derivatives and flavonoids were likely the active constituents of daylily flowers and they conferred a similar extent of neuroprotection, but affected the release of neurotransmitters in a different manner.

Neuroimaging during Trance State: A Contribution to the Study of Dissociation

PubMed Central

Peres, Julio Fernando; Moreira-Almeida, Alexander; Caixeta, Leonardo; Leao, Frederico; Newberg, Andrew

2012-01-01

Despite increasing interest in pathological and non-pathological dissociation, few researchers have focused on the spiritual experiences involving dissociative states such as mediumship, in which an individual (the medium) claims to be in communication with, or under the control of, the mind of a deceased person. Our preliminary study investigated psychography – in which allegedly “the spirit writes through the medium's hand” – for potential associations with specific alterations in cerebral activity. We examined ten healthy psychographers – five less expert mediums and five with substantial experience, ranging from 15 to 47 years of automatic writing and 2 to 18 psychographies per month – using single photon emission computed tomography to scan activity as subjects were writing, in both dissociative trance and non-trance states. The complexity of the original written content they produced was analyzed for each individual and for the sample as a whole. The experienced psychographers showed lower levels of activity in the left culmen, left hippocampus, left inferior occipital gyrus, left anterior cingulate, right superior temporal gyrus and right precentral gyrus during psychography compared to their normal (non-trance) writing. The average complexity scores for psychographed content were higher than those for control writing, for both the whole sample and for experienced mediums. The fact that subjects produced complex content in a trance dissociative state suggests they were not merely relaxed, and relaxation seems an unlikely explanation for the underactivation of brain areas specifically related to the cognitive processing being carried out. This finding deserves further investigation both in terms of replication and explanatory hypotheses. PMID:23166648

Medium-chain fatty acids inhibit mitochondrial metabolism in astrocytes promoting astrocyte-neuron lactate and ketone body shuttle systems.

PubMed

Thevenet, Jonathan; De Marchi, Umberto; Domingo, Jaime Santo; Christinat, Nicolas; Bultot, Laurent; Lefebvre, Gregory; Sakamoto, Kei; Descombes, Patrick; Masoodi, Mojgan; Wiederkehr, Andreas

2016-05-01

Medium-chain triglycerides have been used as part of a ketogenic diet effective in reducing epileptic episodes. The health benefits of the derived medium-chain fatty acids (MCFAs) are thought to result from the stimulation of liver ketogenesis providing fuel for the brain. We tested whether MCFAs have direct effects on energy metabolism in induced pluripotent stem cell-derived human astrocytes and neurons. Using single-cell imaging, we observed an acute pronounced reduction of the mitochondrial electrical potential and a concomitant drop of the NAD(P)H signal in astrocytes, but not in neurons. Despite the observed effects on mitochondrial function, MCFAs did not lower intracellular ATP levels or activate the energy sensor AMP-activated protein kinase. ATP concentrations in astrocytes were unaltered, even when blocking the respiratory chain, suggesting compensation through accelerated glycolysis. The MCFA decanoic acid (300 μM) promoted glycolysis and augmented lactate formation by 49.6%. The shorter fatty acid octanoic acid (300 μM) did not affect glycolysis but increased the rates of astrocyte ketogenesis 2.17-fold compared with that of control cells. MCFAs may have brain health benefits through the modulation of astrocyte metabolism leading to activation of shuttle systems that provide fuel to neighboring neurons in the form of lactate and ketone bodies.-Thevenet, J., De Marchi, U., Santo Domingo, J., Christinat, N., Bultot, L., Lefebvre, G., Sakamoto, K., Descombes, P., Masoodi, M., Wiederkehr, A. Medium-chain fatty acids inhibit mitochondrial metabolism in astrocytes promoting astrocyte-neuron lactate and ketone body shuttle systems. © FASEB.

The Bermuda Triangle of cocaine-induced neuroadaptations.

PubMed

Wolf, Marina E

2010-09-01

Activation of medium spiny neurons (MSNs) of the nucleus accumbens is critical for goal-directed behaviors including cocaine seeking. Studies in cocaine-experienced rodents have revealed three major categories of neuroadaptations that influence the ability of glutamate inputs to activate MSNs: changes in synaptic AMPA receptor levels, changes in extracellular non-synaptic glutamate levels and changes in MSN intrinsic membrane excitability. Most studies have focused on one of these adaptations. This review will consider the possibility that they are causally related and speculate about how time-dependent changes in their interactions may regulate MSN output during early and late withdrawal from repeated cocaine exposure. Copyright 2010 Elsevier Ltd. All rights reserved.

The Bermuda Triangle of Cocaine-Induced Neuroadaptations

PubMed Central

Wolf, Marina E.

2010-01-01

Activation of medium spiny neurons (MSN) of the nucleus accumbens is critical for goal-directed behaviors including cocaine seeking. Studies in cocaine-experienced rodents have revealed three major categories of neuroadaptations that influence the ability of glutamate inputs to activate MSN: changes in synaptic AMPA receptor levels, changes in extracellular non-synaptic glutamate levels, and changes in MSN intrinsic membrane excitability. Most studies have focused on one of these adaptations. Here, we consider the possibility that they are causally related and speculate about how time-dependent changes in their interactions may regulate MSN output during early and late withdrawal from repeated cocaine exposure. PMID:20655604

Redistribution of carbon flux in Torulopsis glabrata by altering vitamin and calcium level.

PubMed

Liu, Liming; Li, Yin; Zhu, Yang; Du, Guocheng; Chen, Jian

2007-01-01

Manipulation of cofactor (thiamine, biotin and Ca(2+)) levels as a potential tool to redistribute carbon flux was studied in Torulopsis glabrata. With sub-optimization of vitamin in fermentation medium, the carbon flux was blocked at the key node of pyruvate, and 69 g/L pyruvate was accumulated. Increasing the concentrations of thiamine and biotin could selectively open the valve of carbon flux from pyruvate to pyruvate dehydrogenase complex, the pyruvate carboxylase (PC) pathway and the channel into the TCA cycle, leading to the over-production of alpha-ketoglutarate. In addition, the activity of PC was enhanced with Ca(2+) present in fermentation medium. By combining high concentration's vitamins and CaCO(3) as the pH buffer, a batch culture was conducted in a 7-L fermentor, with the pyruvate concentration decreased to 21.8 g/L while alpha-ketoglutarate concentration increased to 43.7 g/L. Our study indicated that the metabolic flux could be redistributed to overproduce desired metabolites with manipulating the cofactor levels. Furthermore, the manipulation of vitamin level provided an alternative tool to realize metabolic engineering goals.

The effects of goal variation on adult physical activity behaviour.

PubMed

Moon, Dal-Hyun; Yun, Joonkoo; McNamee, Jeff

2016-10-01

The purposes of this study were to examine the effects of varying levels of goals on increasing daily steps and the frequency of goal achievement among middle-aged adults. Ninety-six adults participated in a randomised control study. Participants were randomly assigned to five different step goal groups: (1) Easy (n = 19), (2) Medium (n = 19), (3) Difficult (n = 19), (4) Do-your-best (n = 19), and (5) No goal (n = 20) based on previous research. The participants wore a pedometer and were asked to reach a pre-established goal during the experimental period. In order to examine the effectiveness of the goal difficulty, (a) an average number of steps taken by different goal conditions and (b) the number of days meeting the assigned goal were tested. A one-way ANCOVA revealed significant step count differences among goal groups. Post hoc analyses indicated that the change in step count in both the Medium and Difficult goal groups was significantly greater than the remaining groups. However, there was no significant difference between the medium and difficult goal conditions. In addition, a one-way ANOVA indicated that there were no significant differences in the frequency of goal achievement among the Easy, Medium, and Difficult goal groups. Results suggest that when promoting physical activity through increasing step counts, researchers and clinicians should design goals that are specific and challenging.

Metabolic Genetic Screens Reveal Multidimensional Regulation of Virulence Gene Expression in Listeria monocytogenes and an Aminopeptidase That Is Critical for PrfA Protein Activation.

PubMed

Friedman, Sivan; Linsky, Marika; Lobel, Lior; Rabinovich, Lev; Sigal, Nadejda; Herskovits, Anat A

2017-06-01

Listeria monocytogenes is an environmental saprophyte and intracellular bacterial pathogen. Upon invading mammalian cells, the bacterium senses abrupt changes in its metabolic environment, which are rapidly transduced to regulation of virulence gene expression. To explore the relationship between L. monocytogenes metabolism and virulence, we monitored virulence gene expression dynamics across a library of genetic mutants grown under two metabolic conditions known to activate the virulent state: charcoal-treated rich medium containing glucose-1-phosphate and minimal defined medium containing limiting concentrations of branched-chain amino acids (BCAAs). We identified over 100 distinct mutants that exhibit aberrant virulence gene expression profiles, the majority of which mapped to nonessential metabolic genes. Mutants displayed enhanced, decreased, and early and late virulence gene expression profiles, as well as persistent levels, demonstrating a high plasticity in virulence gene regulation. Among the mutants, one was noteworthy for its particularly low virulence gene expression level and mapped to an X-prolyl aminopeptidase (PepP). We show that this peptidase plays a role in posttranslational activation of the major virulence regulator, PrfA. Specifically, PepP mediates recruitment of PrfA to the cytoplasmic membrane, a step identified as critical for PrfA protein activation. This study establishes a novel step in the complex mechanism of PrfA activation and further highlights the cross regulation of metabolism and virulence. Copyright © 2017 American Society for Microbiology.

Observations of thunderstorm-related 630 nm airglow depletions

NASA Astrophysics Data System (ADS)

Kendall, E. A.; Bhatt, A.

2015-12-01

The Midlatitude All-sky imaging Network for Geophysical Observations (MANGO) is an NSF-funded network of 630 nm all-sky imagers in the continental United States. MANGO will be used to observe the generation, propagation, and dissipation of medium and large-scale wave activity in the subauroral, mid and low-latitude thermosphere. This network is actively being deployed and will ultimately consist of nine all-sky imagers. These imagers form a network providing continuous coverage over the western United States, including California, Oregon, Washington, Utah, Arizona and Texas extending south into Mexico. This network sees high levels of both medium and large scale wave activity. Apart from the widely reported northeast to southwest propagating wave fronts resulting from the so called Perkins mechanism, this network observes wave fronts propagating to the west, north and northeast. At least three of these anomalous events have been associated with thunderstorm activity. Imager data has been correlated with both GPS data and data from the AIRS (Atmospheric Infrared Sounder) instrument on board NASA's Earth Observing System Aqua satellite. We will present a comprehensive analysis of these events and discuss the potential thunderstorm source mechanism.

Multimedia Didactics for a Knowledge Society

ERIC Educational Resources Information Center

Milan, Matijevic

2008-01-01

It is considered that television as a medium presented a great incentive for change to the global political movement in 1968. In those years, television achieved a notable level of power in terms of its informative, educational and political activity. At the end of the 19th century and the beginning of the 20th century, the communicative power of…

Linear analysis of active-medium two-beam accelerator

NASA Astrophysics Data System (ADS)

Voin, Miron; Schächter, Levi

2015-07-01

We present detailed development of the linear theory of wakefield amplification by active medium and its possible application to a two-beam accelerator (TBA) is discussed. A relativistic train of triggering microbunches traveling along a vacuum channel in an active medium confined by a cylindrical waveguide excites Cherenkov wake in the medium. The wake is a superposition of azimuthally symmetric transverse magnetic modes propagating along a confining waveguide, with a phase velocity equal to the velocity of the triggering bunches. The structure may be designed in such a way that the frequency of one of the modes is close to active-medium resonant frequency, resulting in amplification of the former and domination of a single mode far behind the trigger bunches. Another electron bunch placed in proper phase with the amplified wakefield may be accelerated by the latter. Importantly, the energy for acceleration is provided by the active medium and not the drive bunch as in a traditional TBA. Based on a simplified model, we analyze extensively the impact of various parameters on the wakefield amplification process.

l-Asparaginase from Proteus vulgaris1

PubMed Central

Tosa, Tetsuya; Sano, Ryujiro; Yamamoto, Kozo; Nakamura, Masatoshi; Ando, Katsuko; Chibata, Ichiro

1971-01-01

To produce an immunologically and enzymologically new type of l-asparaginase, 108 strains of bacteria were screened for enzyme production. As a result, 13 bacteria belonging to the genera Alcaligenes, Bacterium, and Proteus were found to produce l-asparaginases in high levels. Among these l-asparaginases, partially purified l-asparaginases from B. cadaveris and P. vulgaris showed antitumor activity. A partially purified l-asparaginase preparation of P. vulgaris did not react with the antibody of Escherichia colil-asparaginase on the Ouchterlony agar plate. Culture conditions for the production of l-asparaginase by P. vulgaris were investigated in detail. The enzyme was produced in high yields when cells were grown aerobically in a medium containing sodium fumarate and corn steep liquor. The addition of glucose or ammonium ion to the medium, however, resulted in depressed production of l-asparaginase. Under the optimum conditions, 3,700 international units of l-asparaginase was obtained from 1 liter of culture medium. Images PMID:5000866

Cross-sectional associations between maternal parenting styles, physical activity and screen sedentary time in children.

PubMed

Van der Geest, K E; Mérelle, S Y M; Rodenburg, G; Van de Mheen, D; Renders, C M

2017-09-29

Children's activity level, including physical activity (PA) and screen sedentary time (SST), is influenced by environmental factors in which parents play a critical role. Different types of parenting styles may influence children's activity level. Inconsistent results were found on the association between parenting styles and PA, and few studies tested the association between parenting styles and SST. This study examined the association between parenting styles, PA and SST and the modifying effect of children's gender and maternal educational level on these associations. Cross-sectional data were collected from parents of children aged 8-11 years old who completed a web-based non-standardized questionnaire (N = 4047). Since 85% of the questionnaires were filled in by mothers, parenting styles are mainly reported by mothers. Multiple linear regression techniques were used to assess the associations between parenting styles (authoritative, permissive, authoritarian and neglectful), and PA and SST (mean min/day). The modifying effect of children's gender and maternal educational level on these associations was explored. P values ≤.0125 were considered as statistically significant based on the Bonferroni correction for four primary analyses. The neglectful parenting style was most widely used (35.3%), while the authoritarian style was least common (14.8%). No significant association was found between parenting styles and PA level. As regards SST, an authoritative parenting style was significantly associated with lower SST in boys while a neglectful parenting style was significantly associated with higher SST in both boys and girls. When the mother had a medium educational level, an authoritative parenting style was significantly associated with lower SST while neglectful parenting was significantly associated with higher SST. No association was found between parenting styles and PA. However, an authoritative parenting style was associated with a reduction in SST and a neglectful parenting style with an increase in SST, especially in boys and in children whose mother had a medium education level. Future studies of parenting practices are needed to gain more insight into the role of parents in children's PA and SST levels, as a basis for the development of interventions tailored to support parents in stimulating PA and reducing SST in children.

Alpha-ketoglutarate enhances freeze-thaw tolerance and prevents carbohydrate-induced cell death of the yeast Saccharomyces cerevisiae.

PubMed

Bayliak, Maria M; Hrynkiv, Olha V; Knyhynytska, Roksolana V; Lushchak, Volodymyr I

2018-01-01

Stress resistance and fermentative capability are important quality characteristics of baker's yeast. In the present study, we examined protective effects of exogenous alpha-ketoglutarate (AKG), an intermediate of the tricarboxylic acid cycle and amino acid metabolism, against freeze-thaw and carbohydrate-induced stresses in the yeast Saccharomyces cerevisiae. Growth on AKG-supplemented medium prevented a loss of viability and improved fermentative capacity of yeast cells after freeze-thaw treatment. The cells grown in the presence of AKG had higher levels of amino acids (e.g., proline), higher metabolic activity and total antioxidant capacity, and higher activities of catalase, NADP-dependent glutamate dehydrogenase and glutamine synthase compared to control ones. Both synthesis of amino acids and enhancement of antioxidant system capacity could be involved in AKG-improved freeze-thaw tolerance in S. cerevisiae. Cell viability dramatically decreased under incubation of stationary-phase yeast cells in 2% glucose or fructose solutions (in the absence of the other nutrients) as compared with incubation in distilled water or in 10 mM AKG solution. The decrease in cell viability was accompanied by acidification of the medium, and decrease in cellular respiration, aconitase activity, and levels of total protein and free amino acids. The supplementation with 10 mM AKG effectively prevented carbohydrate-induced yeast death. Protective mechanisms of AKG could be associated with the intensification of respiration and prevention of decreasing protein level as well as with direct antioxidant AKG action.

Physical Activity of Malaysian Primary School Children: Comparison by Sociodemographic Variables and Activity Domains.

PubMed

Wong, Jyh Eiin; Parikh, Panam; Poh, Bee Koon; Deurenberg, Paul

2016-07-01

This study describes the physical activity of primary school children according to sociodemographic characteristics and activity domains. Using the Malaysian South East Asian Nutrition Surveys data, 1702 children aged 7 to 12 years were included in the analysis. Physical activity was reported as a total score and categorized into low, medium, and high levels based on Physical Activity Questionnaire for Older Children. Higher overall activity scores were found in boys, younger age, non-Chinese ethnicity, and normal body mass index category. Sex, age, and ethnicity differences were found in structured or organized, physical education, and outside-of-school domain scores. Transport-related scores differed by age group, ethnicity, household income, and residential areas but not among the three physical activity levels. Participation of girls, Chinese, and older children were low in overall and almost all activity domains. Sociodemographic characteristics are important factors to consider in increasing the different domains of physical activity among Malaysian children. © 2016 APJPH.

Resorption behavior of a nanostructured bone substitute: in vitro investigation and clinical application.

PubMed

Reichert, Christoph; Götz, Werner; Reimann, Susanne; Keilig, Ludger; Hagner, Martin; Bourauel, Christoph; Jäger, Andreas

2013-03-01

To develop an in vitro assay for quantitative analysis of the degradation to which a bone substitute is exposed by osteoclasts. The aim of establishing this method was to improve the predictability of carrying out tooth movements via bone substitutes and to provide a basis for verification in exemplary clinical cases. After populating a bone substitute (NanoBone®; ArtOss, Germany) with osteoclastic cells, inductively-coupled mass spectrometry was used to evaluate changing calcium levels in the culture medium as a marker of resorption activity. It was observed that calcium levels increased substantially in the culture medium with the cells populating the bone substitute. This in vitro assay is a valid method that can assist clinicians in selecting the appropriate materials for certain patients. While tooth movements occurring through this material were successful, uncertainty about the approach will remain as long-term results are not available.

Fission fragment excited laser system

DOEpatents

McArthur, David A.; Tollefsrud, Philip B.

1976-01-01

A laser system and method for exciting lasing action in a molecular gas lasing medium which includes cooling the lasing medium to a temperature below about 150 K and injecting fission fragments through the lasing medium so as to preferentially excite low lying vibrational levels of the medium and to cause population inversions therein. The cooled gas lasing medium should have a mass areal density of about 5 .times. 10.sup.-.sup.3 grams/square centimeter, relaxation times of greater than 50 microseconds, and a broad range of excitable vibrational levels which are excitable by molecular collisions.

Mevalonate kinase activity during different stages of plant regeneration from nodular callus cultures in white pine (Pinus strobus).

PubMed

Tang, Wei; Newton, Ronald J

2006-02-01

Mevalonate kinase (MK) catalyzes a step in the isoprenoid biosynthetic pathway, which leads to a huge number of compounds that play important roles in plant growth and development. Here, we report on changes in MK activity in white pine (Pinus strobus L.) during plant regeneration by adventitious shoot organogenesis from cotyledons of mature embryos, including nodular callus induction, shoot formation and rooting. Nodular calli were induced from Pinus strobus (PS) embryos by culture in nodular callus induction medium in a 0-, 8- or 16-h photoperiod. Mevalonate kinase activity peaked in nodular calli after three weeks of culture on nodular callus induction medium in a 16-h photoperiod, whereas frequency of nodular callus formation peaked after 4 weeks of culture on nodular callus induction medium in darkness. During adventitious shoot formation, MK activity peaked in shoots derived from dark-grown nodular calli after 3 weeks on bud formation medium, and frequency of shoot formation was highest in dark-grown nodular calli cultured on bud formation medium for 4 weeks. During rooting, MK activity peaked 2 weeks after transfer of adventitious shoots to rooting medium and rooting frequency was highest in adventitious shoots after 3 weeks on rooting medium. Although during nodular callus induction in darkness MK activity was inversely related to frequency of nodular callus formation, MK activity was highly correlated with frequency of shoot formation and with rooting frequency. The observed increase in MK activity preceding rooting suggests that MK could serve as a marker for rooting of white pine shoots in vitro.

Differential cellular responses by oncogenic levels of c-Myc expression in long-term confluent retinal pigment epithelial cells.

PubMed

Wang, Yiping; Cheng, Xiangdong; Samma, Muhammad Kaleem; Kung, Sam K P; Lee, Clement M; Chiu, Sung Kay

2018-06-01

c-Myc is a highly pleiotropic transcription factor known to control cell cycle progression, apoptosis, and cellular transformation. Normally, ectopic expression of c-Myc is associated with promoting cell proliferation or triggering cell death via activating p53. However, it is not clear how the levels of c-Myc lead to different cellular responses. Here, we generated a series of stable RPE cell clones expressing c-Myc at different levels, and found that consistent low level of c-Myc induced cellular senescence by activating AP4 in post-confluent RPE cells, while the cells underwent cell death at high level of c-Myc. In addition, high level of c-Myc could override the effect of AP4 on cellular senescence. Further knockdown of AP4 abrogated senescence-like phenotype in cells expressing low level of c-Myc, and accelerated cell death in cells with medium level of c-Myc, indicating that AP4 was required for cellular senescence induced by low level of c-Myc.

Stimulation of insulin secretion by medium-chain triglycerides in patients with cirrhosis 1

PubMed Central

McCullough, Frank S.; Tzagournis, Manuel; Greenberger, Norton J.; Linscheer, Willem G.

1971-01-01

Oral medium-chain triglycerides were given to 10 normal volunteers, 12 cirrhotics (group I) without and 28 cirrhotics (group II) with abnormal portal systemic communications (ascites, splenomegaly, oesophageal varices, or surgically-created portacaval shunts). After 30 ml of medium-chain triglyceride oil there was no appreciable change in serum glucose levels in any of the three groups nor in serum insulin levels in the normals and in cirrhotics in group I. However, there was a significant increase in serum insulin levels in the cirrhotic patients in group II. It is suggested that the rise in serum insulin levels after medium-chain triglycerides noted in the cirrhotics with shunts is due to shunting of insulin-containing portal blood around the liver (anatomical shunts) and to a diminished hepatic cell mass capable of extracting insulin (functional shunt). This differential response of serum insulin levels to medium-chain triglycerides may prove to be of value in detecting the presence of abnormal portal systemic communications in cirrhotic patients. PMID:5548559

Effects of temperature and medium composition on inhibitory activities of gossypol-related compounds against aflatoxigenic fungi.

PubMed

Mellon, J E; Dowd, M K; Beltz, S B

2013-07-01

To investigate the effects of temperature and medium composition on growth/aflatoxin inhibitory activities of terpenoids gossypol, gossypolone and apogossypolone against Aspergillus flavus and A. parasiticus. The compounds were tested at a concentration of 100 μg ml(-1) in a Czapek Dox (Czapek) agar medium at 25, 31 and 37°C. Increased incubation temperature marginally increased growth inhibition caused by these compounds, but reduced the aflatoxin inhibition effected by gossypol. Gossypolone and apogossypolone retained good aflatoxin inhibitory activity against A. flavus and A. parasiticus at higher incubation temperatures. However, increased temperature also significantly reduced aflatoxin production in control cultures. The effects of the terpenoids on fungal growth and aflatoxin production against the same fungi were also determined in Czapek, Czapek with a protein/amino acid addendum and yeast extract sucrose (YES) media. Growth of these fungi in the protein-supplemented Czapek medium or in the YES medium greatly reduced the growth inhibition effects of the terpenoids. Apogossypolone displayed strong anti-aflatoxigenic activity in the Czapek medium, but this activity was significantly reduced in the protein-amended Czapek and YES media. Gossypol, which displayed little to no aflatoxin inhibitory activity in the Czapek medium, did yield significant anti-aflatoxigenic activity in the YES medium. Incubation temperature and media composition are important parameters involved in the regulation of aflatoxin production in A. flavus and A. parasiticus. These parameters also affect the potency of growth and aflatoxin inhibitory activities of these gossypol-related compounds against aflatoxigenic fungi. Studies utilizing gossypol-related compounds as inhibitory agents of biological activities should be interpreted with caution due to compound interaction with multiple components of the test system, especially serum proteins. Published [2013]. This article is a U.S. Government work and is in the public domain in the USA.

Dependence of synchronized bursting activity on medium stirring and the perfusion rate in a cultured network of neurons

NASA Astrophysics Data System (ADS)

Heo, Ryoun; Kim, Hyun; Lee, Kyoung J.

2016-05-01

A cultured network of neurons coupled with a multi-electrode-array (MEA) recording system has been a useful platform for investigating various issues in neuroscience and engineering. The neural activity supported by the system can be sensitive to environmental fluctuations, for example, in the medium's nutrient composition, ph, and temperature, and to mechanical disturbances, yet this issue has not been the subject. Especially, a normal practice in maintaining neuronal cell cultures involves an intermittent sequence of medium exchanges, typically at a time interval of a few days, and one such sudden medium exchange is unavoidably accompanied by many unintended disturbances. Here, based on a quantitative time-series analysis of synchronized bursting events, we explicitly demonstrate that such a medium exchange can, indeed, bring a huge change in the existing neural activity. Subsequently, we develop a medium perfusion-stirring system and an ideal protocol that can be used in conjunction with a MEA recording system, providing long-term stability. Specifically, we systematically evaluate the effects of medium stirring and perfusion rates. Unexpectedly, even some vigorous mechanical agitations do not have any impacts on neural activity. On the other hand, too much replenishment ( e.g., 1.8 ml/day for a 1.8-ml dish) of neurobasal medium results in an excitotoxicity.

Replacement of alfalfa hay (Medicago sativa L.) with subabul (Leucaena leucocephala) leaf meal in diets of Najdi goats: effect on digestion activity of rumen microorganisms.

PubMed

Mohammadabadi, Tahereh; Jolazadeh, Alireza

2017-08-01

This study investigated the effect of replacing alfalfa hay by subabul leaf meal (SLM) on digestion, fermentation parameters and rumen bacteria and fungi activity of Najdi goats. Six Najdi goats (150 ± 15 days of age and initial body weight of 35 ± 1.1 kg) were randomly assigned to one of two dietary treatments in a balanced completely randomized design (three goats per treatment) for 56 days. Experimental treatments included alfalfa hay as control group and diet containing SLM (SLM replacing alfalfa hay at 50% level). Bacterial and fungi activity and rumen fermentation parameters of animals fed experimental diets were determined. Dry matter disappearance (DMD) was unaffected by replacing SLM with alfalfa hay for both rumen bacteria and fungi in different incubation times, except for 48 h of incubation in specific culture medium of mixed rumen bacteria, which decreased for SLM group (P > 0.05). NDF disappearance (NDFD) and ADF disappearance (ADFD) after 24 and 48 h of incubation in specific culture medium of mixed rumen bacteria was not affected by experimental diets (P > 0.05). However, 72 h after incubation, NDFD in SLM treatment decreased (P > 0.05). Gas production parameters of rumen bacteria were similar for both experimental diets, but partitioning factor (PF), efficiency microbial biomass production (EMBP), microbial protein production (MP), and organic matter truly digested (OMTD) decreased (p < 0.05) in specific culture medium of mixed rumen fungi for SLM diet. There was significant decrease in total protozoa population for diet containing SLM (p < 0.05). Results indicated that replacement of alfalfa hay by SLM had no major effect on rumen microorganisms' activity of Najdi goats, so it may be used as an alternative for alfalfa (at 50% level) in susceptible areas.

Histamine Induces Bovine Rumen Epithelial Cell Inflammatory Response via NF-κB Pathway.

PubMed

Sun, Xudong; Yuan, Xue; Chen, Liang; Wang, Tingting; Wang, Zhe; Sun, Guoquan; Li, Xiaobing; Li, Xinwei; Liu, Guowen

2017-01-01

Subacute ruminal acidosis (SARA) is a common disease in high-producing lactating cows. Rumenitis is the initial insult of SARA and is associated with the high concentrations of histamine produced in the rumen of dairy cows during SARA. However, the exact mechanism remains unclear. The objective of the current study is to investigate whether histamine induces inflammation of rumen epithelial cells and the underlying mechanism of this process. Bovine rumen epithelial cells were cultured and treated with different concentrations of histamine and pyrrolidine dithiocarbamate (PDTC, an NF-κB inhibitor) cultured in different pH medium (pH 7.2 or 5.5). qRT-PCR, Western-blotting, ELISA and immunocytofluorescence were used to evaluate whether histamine activated the NF-κB pathway and inflammatory cytokines. The results showed that histamine significantly increased the activity of IKK β and the phosphorylation levels of IκB α, as well as upregulated the mRNA and protein expression levels of NF-κB p65 in the rumen epithelial cells cultured in neutral (pH=7.2) and acidic (pH=5.5) medium. Furthermore, histamine treatment also significantly increased the transcriptional activity of NF-κB p65. High expression and transcriptional activity of NF-κB p65 significantly increased the mRNA expressions and concentrations of inflammatory cytokines, tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and interleukin 1 beta (IL-1β), thereby inducing the inflammatory response in bovine rumen epithelial cells. However, inhibition of NF-κB p65 by PDTC significantly decreased the expressions and concentrations of the inflammatory cytokines induced by histamine in the rumen epithelial cells cultured in the neutral and acidic medium. The present data indicate that histamine induces the inflammatory response of bovine rumen epithelial cells through the NF-κB pathway. © 2017 The Author(s). Published by S. Karger AG, Basel.

Enhancing absorption in coated semiconductor nanowire/nanorod core-shell arrays using active host matrices

NASA Astrophysics Data System (ADS)

Jule, Leta; Dejene, Francis; Roro, Kittessa

2016-12-01

In the present work, we investigated theoretically and experimentally the interaction of radiation field phenomena interacting with arrays of nanowire/nanorod core-shell embedded in active host matrices. The optical properties of composites are explored including the case when the absorption of propagating wave by dissipative component is completely compensated by amplification in active (lasing) medium. On the basis of more elaborated modeling approach and extended effective medium theory, the effective polarizability and the refractive index of electromagnetic mode dispersion of the core-shell nanowire arrays are derived. ZnS(shell)-coated by sulphidation process on ZnO(shell) nanorod arrays grown on (100) silicon substrate by chemical bath deposition (CBD) has been used for theoretical comparison. Compared with the bare ZnO nanorods, ZnS-coated core/shell nanorods exhibit a strongly reduced ultraviolet (UV) emission and a dramatically enhanced deep level (DL) emission. Obviously, the UV and DL emission peaks are attributed to the emissions of ZnO nanorods within ZnO/ZnS core/shell nanorods. The reduction of UV emission after ZnS coating seems to agree with the charge separation mechanism of type-II band alignment that holes transfer from the core to shell, which would quench the UV emission to a certain extent. Our theoretical calculations and numerical simulation demonstrate that the use of active host (amplifying) medium to compensate absorption at metallic inclusions. Moreover the core-shell nanorod/nanowire arrays create the opportunity for broad band absorption and light harvesting applications.

Mesenchymal stem cell-conditioned medium triggers neuroinflammation and reactive species generation in organotypic cultures of rat hippocampus.

PubMed

Horn, Ana Paula; Bernardi, Andressa; Luiz Frozza, Rudimar; Grudzinski, Patrícia Bencke; Hoppe, Juliana Bender; de Souza, Luiz Fernando; Chagastelles, Pedro; de Souza Wyse, Angela Terezinha; Bernard, Elena Aida; Battastini, Ana Maria Oliveira; Campos, Maria Martha; Lenz, Guido; Nardi, Nance Beyer; Salbego, Christianne

2011-07-01

Cell therapy using bone marrow-derived mesenchymal stem cells (MSCs) seems to be a new alternative for the treatment of neurodegenerative diseases. Despite several promising results with their use, possible side effects are still unknown. In a previous work, we have shown that MSC-conditioned medium is toxic to hippocampal slice cultures and aggravates cell death induced by oxygen and glucose deprivation. In this work, we investigated whether the inflammatory response and/or reactive species formation could be involved in that toxicity. Rat organotypic hippocampal cultures were exposed for 24 h to conditioned medium from MSCs isolated from rat bone marrow. A marked glial activation was observed after exposure of cultures to MSC-conditioned medium, as evidenced by glial fibrillary acid protein (GFAP) and isolectin B(4) increase. Tumor necrosis factor-α and interleukin-6 levels were increased in the culture medium, and 2,7-dihydrodichlorofluorescein diacetate oxidation (indicating reactive species generation) and inducible nitric oxide synthase (iNOS) immunocontent were also higher after exposure of cultures to MSC-conditioned medium. Antioxidants (ascorbic acid and TROLOX(®)), N(ω)-nitro-l-arginine methyl ester hydrochloride, and anti-inflammatory drugs (indomethacin and dexamethasone) reduced cell death in hippocampal organotypic cultures after their exposure to MSC-conditioned medium. The results obtained here suggest that MSC-secreted factors trigger reactive species generation and neuroinflammation in organotypic cultures of hippocampus, introducing a note of caution in the use of these cells for neurological application.

Effects of neural stem cell media on hypoxic injury in rat hippocampal slice cultures.

PubMed

Lee, Na Mi; Chae, Soo Ahn; Lee, Hong Jun

2017-12-15

Neonatal hypoxic-ischemic brain injuries cause serious neurological sequelae, yet there is currently no effective treatment for them. We hypothesized that neurotrophic factors released into the medium by stem cells could supply hypoxia-damaged organotypic hippocampal slice cultures with regenerative abilities. We prepared organotypic slice cultures of the hippocampus of 7-day-old Sprague-Dawley rats based on the modified Stoppini method; slices were cultured for 14days in vitro using either Gahwiler's medium (G-medium) or stem cell-conditioned medium (S-medium) as culture medium. At day 14 in vitro, hippocampal slice cultures were exposed to 95% N 2 and 5% CO 2 for 3h to induce hypoxic damage, the extent of which was then measured using propidium iodide fluorescence and immunohistochemistry images. We performed dot blotting to estimate neurotrophic/growth factor levels in the G- and S-media. Organotypic hippocampal slices cultured using S-medium after hypoxic injury were significantly less damaged than those cultured using G-medium. GLUT1, NGF, GDNF, VEGF, GCSF, and IGF2 levels were higher in S-medium than in G-medium, whereas FGF1, HIF, and MCP3 levels were not significantly different between media. In conclusion, we found that stem cell-conditioned medium had a neuroprotective effect against hypoxic injury, and that, of the various neurotrophic factors in S-medium, NGF, GDNF, and VEGF can contribute to neuroprotection. Copyright © 2017 Elsevier B.V. All rights reserved.

Glycine Betaine, Carnitine, and Choline Enhance Salinity Tolerance and Prevent the Accumulation of Sodium to a Level Inhibiting Growth of Tetragenococcus halophila

PubMed Central

Robert, Hervé; Le Marrec, Claire; Blanco, Carlos; Jebbar, Mohamed

2000-01-01

Natural-abundance 13C-nuclear magnetic resonance was used to probe the intracellular organic solute content of the moderately halophilic bacterium Tetragenococcus halophila. When grown in complex growth media supplemented or not with NaCl, T. halophila accumulates glycine betaine and carnitine. Unlike other moderate halophiles, T. halophila was not able to produce potent osmoprotectants (such as ectoines and glycine betaine) through de novo synthesis when cultured in defined medium under hyperosmotic constraint. Addition of 2 mM carnitine, glycine betaine, or choline to defined medium improved growth parameters, not only at high salinity (up to 2.5 M NaCl) but also in media lacking NaCl. These compounds were taken up when available in the surrounding medium. The transport activity occurred at low and high salinities and seems to be constitutive. Glycine betaine and carnitine were accumulated by T. halophila in an unmodified form, while exogenously provided choline led to an intracellular accumulation of glycine betaine. This is the first evidence of the existence of a choline-glycine betaine pathway in a lactic acid bacterium. An assay showed that the compatible solutes strikingly repressed the accumulation of glutamate and slightly increased the intracellular potassium level only at high salinity. Interestingly, osmoprotectant-treated cells were able to maintain the intracellular sodium concentration at a relatively constant level (200 to 300 nmol/mg [dry weight]), independent of the NaCl concentration of the medium. In contrast, in the absence of osmoprotectant, the intracellular sodium content increased sharply from 200 to 2,060 nmol/mg (dry weight) when the salinity of the medium was raised from 1 to 2 M. Indeed, the imported compatible solutes play an actual role in regulating the intracellular Na+ content and confer a much higher salt tolerance to T. halophila. PMID:10653711

Fully Dedifferentiated Chondrocytes Expanded in Specific Mesenchymal Stem Cell Growth Medium with FGF2 Obtains Mesenchymal Stem Cell Phenotype In Vitro but Retains Chondrocyte Phenotype In Vivo

PubMed Central

Lee, Jungsun; Lee, Jin-Yeon; Chae, Byung-Chul; Jang, Jeongho

2017-01-01

Given recent progress in regenerative medicine, we need a means to expand chondrocytes in quantity without losing their regenerative capability. Although many reports have shown that growth factor supplementation can have beneficial effects, the use of growth factor–supplemented basal media has widespread effect on the characteristics of chondrocytes. Chondrocytes were in vitro cultured in the 2 most widely used chondrocyte growth media, conventional chondrocyte culture medium and mesenchymal stem cell (MSC) culture medium, both with and without fibroblast growth factor-2 (FGF2) supplementation. Their expansion rates, expressions of extracellular matrix–related factors, senescence, and differentiation potentials were examined in vitro and in vivo. Our results revealed that chondrocytes quickly dedifferentiated during expansion in all tested media, as assessed by the loss of type II collagen expression. The 2 basal media (chondrocyte culture medium vs. MSC culture medium) were associated with distinct differences in cell senescence. Consistent with the literature, FGF2 was associated with accelerated dedifferentiation during expansion culture and superior redifferentiation upon induction. However, chondrocytes expanded in FGF2-containing conventional chondrocyte culture medium showed MSC-like features, as indicated by their ability to direct ectopic bone formation and cartilage formation. In contrast, chondrocytes cultured in FGF2-supplemented MSC culture medium showed potent chondrogenesis and almost no bone formation. The present findings show that the chosen basal medium can exert profound effects on the characteristics and activity of in vitro–expanded chondrocytes and indicate that right growth factor/medium combination can help chondrocytes retain a high-level chondrogenic potential without undergoing hypertrophic transition. PMID:29251111

DCEBIO facilitates myogenic differentiation via intermediate conductance Ca2+ activated K+ channel activation in C2C12 myoblasts.

PubMed

Tanaka, Shoko; Ono, Yuko; Sakamoto, Kazuho

2017-04-01

Membrane hyperpolarization is suggested to be a trigger for skeletal muscle differentiation. We investigated whether DCEBIO, an opener of the small/intermediate conductance Ca 2+ activated K + (SK Ca /IK Ca ) channels, increase myogenic differentiation in C2C12 skeletal myoblasts. DCEBIO significantly increased myotube formation, protein expression level of myosin heavy chain II, and mRNA expression level of myogenin in C2C12 myoblasts cultured in differentiation medium. DCEBIO induced myotube formation and hyperpolarization were reduced by the IK Ca channel blocker TRAM-34, but not by the SK Ca channel blocker apamin. These findings show that DCEBIO increases myogenic differentiation by activating IK Ca channels. Copyright © 2017 The Authors. Production and hosting by Elsevier B.V. All rights reserved.

Inorganic polyphosphate in the yeast Saccharomyces cerevisiae with a mutation disturbing the function of vacuolar ATPase.

PubMed

Tomaschevsky, A A; Ryasanova, L P; Kulakovskaya, T V; Kulaev, I S

2010-08-01

A mutation in the vma2 gene disturbing V-ATPase function in the yeast Saccharomyces cerevisiae results in a five- and threefold decrease in inorganic polyphosphate content in the stationary and active phases of growth on glucose, respectively. The average polyphosphate chain length in the mutant cells is decreased. The mutation does not prevent polyphosphate utilization during cultivation in a phosphate-deficient medium and recovery of its level on reinoculation in complete medium after phosphate deficiency. The content of short chain acid-soluble polyphosphates is recovered first. It is supposed that these polyphosphates are less dependent on the electrochemical gradient on the vacuolar membrane.

Effects of intake level of a mixed diet on chewing activity and on particle size of ruminated boli, ruminal digesta fractions and faeces of steers.

PubMed

Kovács, P L; Südekum, K H; Stangassinger, M

1997-01-01

This study evaluated the effects of intake of a mixed diet on chewing activity during eating and rumination and the relationship between the chewing activity and the particle size of the ruminated boli, ruminal digesta fractions and faeces in steers. Six ruminally cannulated steers received a mixed forage/concentrate diet (68:32, dry matter basis). The diet was offered twice daily at approximately 1, 1.5 and 2 times the estimated maintenance energy requirements (low, medium and high intake, respectively) in a repeated 3 x 3 Latin square design. The rumens were emptied manually and samples of the ruminated boli and of the ruminal upper strata were collected at four different times throughout the day. The dry matter weight distribution of the total amount of recovered particles was determined by a wet-sieving procedure. Numerically, the effect of intake on the mean particle sizes of the different materials was small. However, the mean particle size was reduced by almost nine tenths from their size at intake of the mixed diet (4.78 mm) to defecation (0.51 mm). The total number of minutes chewing and eating and ruminating increased as the intake level increased. When related to 1 kg of dry matter intake, only the eating and chewing times were significantly longer for the high as compared to the medium intake. Rumination patterns were examined using a cosinor model. Data indicated that the average amount of time spent ruminating also increased as the intake level increased. The overall pattern of rumination was not impaired by higher intake levels. The amount of large (> or = 4 mm) particles that escaped per minute of rumination time between 3 and 7.5 h postfeeding was similar for all the intake levels. It was concluded that an active breakdown process occurred in the rumen which could cope with the higher intake levels of the mixed diet over the range of 1 to 2 times the maintenance energy requirement level.

Medium-chain triglyceride ameliorates insulin resistance and inflammation in high fat diet-induced obese mice.

PubMed

Geng, Shanshan; Zhu, Weiwei; Xie, Chunfeng; Li, Xiaoting; Wu, Jieshu; Liang, Zhaofeng; Xie, Wei; Zhu, Jianyun; Huang, Cong; Zhu, Mingming; Wu, Rui; Zhong, Caiyun

2016-04-01

The aim of the present study was to investigate the in vivo effects of dietary medium-chain triglyceride (MCT) on inflammation and insulin resistance as well as the underlying potential molecular mechanisms in high fat diet-induced obese mice. Male C57BL/6J mice (n = 24) were fed one of the following three diets for a period of 12 weeks: (1) a modified AIN-76 diet with 5 % corn oil (normal diet); (2) a high-fat control diet (17 % w/w lard and 3 % w/w corn oil, HFC); (3) an isocaloric high-fat diet supplemented with MCT (17 % w/w MCT and 3 % w/w corn oil, HF-MCT). Glucose metabolism was evaluated by fasting blood glucose levels and intraperitoneal glucose tolerance test. Insulin sensitivity was evaluated by fasting serum insulin levels and the index of homeostasis model assessment-insulin resistance. The levels of serum interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor-α were measured by ELISA, and hepatic activation of nuclear factor κB (NF-κB) and mitogen-activated protein kinase (MAPK) pathways was determined using western blot analysis. Compared to HFC diet, consumption of HF-MCT did not induce body weight gain and white adipose tissue accumulation in mice. HFC-induced increases in serum fasting glucose and insulin levels as well as glucose intolerance were prevented by HF-MCT diet. Meanwhile, HF-MCT resulted in significantly lower serum IL-6 level and higher IL-10 level, and lower expression levels of inducible nitric oxide synthase and cyclooxygenase-2 protein in liver tissues when compared to HFC. In addition, HF-MCT attenuated HFC-triggered hepatic activation of NF-κB and p38 MAPK. Our study demonstrated that MCT was efficacious in suppressing body fat accumulation, insulin resistance, inflammatory response, and NF-κB and p38 MAPK activation in high fat diet-fed mice. These data suggest that MCT may exert beneficial effects against high fat diet-induced insulin resistance and inflammation.

Extracellular cholesterol oxidase production by Streptomyces aegyptia, in vitro anticancer activities against rhabdomyosarcoma, breast cancer cell-lines and in vivo apoptosis.

PubMed

El-Naggar, Noura El-Ahmady; Soliman, Hoda M; El-Shweihy, Nancy M

2018-02-09

In recent years, microbial cholesterol oxidases have gained great attention due to its widespread use in medical applications for serum cholesterol determination. Streptomyces aegyptia strain NEAE-102 exhibited high level of extracellular cholesterol oxidase production using a minimum medium containing cholesterol as the sole source of carbon. Fifteen variables were screened using Plackett-Burman design for the enhanced cholesterol oxidase production. The most significant variables affecting enzyme production were further optimized by using the face-centered central composite design. The statistical optimization resulted in an overall 4.97-fold increase (15.631 UmL -1 ) in cholesterol oxidase production in the optimized medium as compared with the unoptimized medium before applying Plackett Burman design (3.1 UmL -1 ). The purified cholesterol oxidase was evaluated for its in vitro anticancer activities against five human cancer cell lines. The selectivity index values on rhabdomyosarcoma and breast cancer cell lines were 3.26 and 2.56; respectively. The in vivo anticancer activity of cholesterol oxidase was evaluated against Ehrlich solid tumor model. Compared with control mice, tumors growth was significantly inhibited in the mice injected with cholesterol oxidase alone, doxorubicin alone and cholesterol oxidase/doxorubicin combination by 60.97%, 72.99% and 97.04%; respectively. These results demonstrated that cholesterol oxidase can be used as a promising natural anticancer drug.

Endometrial nitric oxide synthase activity in mares susceptible or resistant to persistent breeding-induced endometritis and the effect of a specific iNOS inhibitor in vitro.

PubMed

Khan, F A; Chenier, T S; Foster, R A; Hewson, J; Scholtz, E L

2018-06-01

Emerging research suggests that the nitric oxide system may play a role in persistent breeding-induced endometritis (PBIE) in the mare. Differences in uterine nitric oxide (NO) levels between mares susceptible or resistant to PBIE and a dose-dependent inhibitory effect of NO on uterine contractility have been demonstrated. The objectives of this study were to investigate the difference in total nitric oxide synthase (NOS) activity of the endometrium between susceptible and resistant mares and the effect of a specific inducible nitric oxide synthase (iNOS) inhibitor on the endometrial NOS activity in vitro. Six susceptible and six resistant mares were selected based on preset criteria and the results of an intrauterine challenge with killed spermatozoa during oestrus. Endometrial biopsy samples were collected 24 hr post-challenge and cultured at 37°C for 24 hr in L-arginine supplemented minimum essential medium with or without a specific iNOS inhibitor (1,400 W dihydrochloride, 1 mM). The medium and the cultured endometrial tissue were collected after 24 hr of culture and assayed for NO and total protein, respectively. Total NO content of the medium, normalized to endometrial tissue wet weight or total protein, was used as a measure of endometrial NOS activity. Non-parametric tests were applied for statistical analysis. Susceptible mares had significantly greater endometrial NOS activity than resistant mares. The iNOS inhibitor treatment significantly reduced NOS activity in endometrial samples derived from susceptible and resistant mares. These findings provide a basis for in vivo testing of specific iNOS inhibitors as preventative or therapeutic options for PBIE in mares. © 2018 Blackwell Verlag GmbH.

Promoting Physical Activity among Sedentary Women Using Pedometers

ERIC Educational Resources Information Center

Sidman, Cara L.; Corbin, Charles B.; Le Masurier, Guy

2004-01-01

Sedentary women (n=92) classified as low (L), medium (M), and high (H) in baseline step counts and assigned to 10,000-step goal (TSG) and personal step goal (PSG) groups (within levels) were compared on goal attainment and step counts. A significant interaction for goal attainment, F(2, 86)=4.51, p=.014, indicated that the L group was…

Processes and Their Products: A Comparison of Task Sequences and Outcome in EAP Writing Classes.

ERIC Educational Resources Information Center

Allison, Desmond; And Others

1995-01-01

A recent study proposed a rational for investigating the effects of mediating tasks upon a summary writing activity for tertiary-level students using English as an educational medium. The tasks in this study involved a group discussion or a reading comprehension exercise, with a third condition providing for immediate access to the text and…

Water quality assessment of Australian ports using water quality evaluation indices

PubMed Central

Jahan, Sayka

2017-01-01

Australian ports serve diverse and extensive activities, such as shipping, tourism and fisheries, which may all impact the quality of port water. In this work water quality monitoring at different ports using a range of water quality evaluation indices was applied to assess the port water quality. Seawater samples at 30 stations in the year 2016–2017 from six ports in NSW, Australia, namely Port Jackson, Botany, Kembla, Newcastle, Yamba and Eden, were investigated to determine the physicochemical and biological variables that affect the port water quality. The large datasets obtained were designed to determine the Water Quality Index, Heavy metal Evaluation Index, Contamination Index and newly developed Environmental Water Quality Index. The study revealed medium water quality index and high and medium heavy metal evaluation index at three of the study ports and high contamination index in almost all study ports. Low level dissolved oxygen and higher level of total dissolved solids, turbidity, fecal coliforms, copper, iron, lead, zinc, manganese, cadmium and cobalt are mainly responsible for the poor water qualities of the port areas. Good water quality at the background samples indicated that various port activities are the likely cause for poor water quality inside the port area. PMID:29244876

Quantitative evaluation of waste prevention on the level of small and medium sized enterprises (SMEs).

PubMed

Laner, David; Rechberger, Helmut

2009-02-01

Waste prevention is a principle means of achieving the goals of waste management and a key element for developing sustainable economies. Small and medium sized enterprises (SMEs) contribute substantially to environmental degradation, often not even being aware of their environmental effects. Therefore, several initiatives have been launched in Austria aimed at supporting waste prevention measures on the level of SMEs. To promote the most efficient projects, they have to be evaluated with respect to their contribution to the goals of waste management. It is the aim of this paper to develop a methodology for evaluating waste prevention measures in SMEs based on their goal orientation. At first, conceptual problems of defining and delineating waste prevention activities are briefly discussed. Then an approach to evaluate waste prevention activities with respect to their environmental performance is presented and benchmarks which allow for an efficient use of the available funds are developed. Finally the evaluation method is applied to a number of former projects and the calculated results are analysed with respect to shortcomings and limitations of the model. It is found that the developed methodology can provide a tool for a more objective and comprehensible evaluation of waste prevention measures.

Prolonged Fasting Identifies Heat Shock Protein 10 as a Sirtuin 3 Substrate

PubMed Central

Lu, Zhongping; Chen, Yong; Aponte, Angel M.; Battaglia, Valentina; Gucek, Marjan; Sack, Michael N.

2015-01-01

Although Sirtuin 3 (SIRT3), a mitochondrially enriched deacetylase and activator of fat oxidation, is down-regulated in response to high fat feeding, the rate of fatty acid oxidation and mitochondrial protein acetylation are invariably enhanced in this dietary milieu. These paradoxical data implicate that additional acetylation modification-dependent levels of regulation may be operational under nutrient excess conditions. Because the heat shock protein (Hsp) Hsp10-Hsp60 chaperone complex mediates folding of the fatty acid oxidation enzyme medium-chain acyl-CoA dehydrogenase, we tested whether acetylation-dependent mitochondrial protein folding contributes to this regulatory discrepancy. We demonstrate that Hsp10 is a functional SIRT3 substrate and that, in response to prolonged fasting, SIRT3 levels modulate mitochondrial protein folding. Acetyl mutagenesis of Hsp10 lysine 56 alters Hsp10-Hsp60 binding, conformation, and protein folding. Consistent with Hsp10-Hsp60 regulation of fatty acid oxidation enzyme integrity, medium-chain acyl-CoA dehydrogenase activity and fat oxidation are elevated by Hsp10 acetylation. These data identify acetyl modification of Hsp10 as a nutrient-sensing regulatory node controlling mitochondrial protein folding and metabolic function. PMID:25505263

Antibiotics in the human food chain: establishing no effect levels of tetracycline, neomycin, and erythromycin using a chemostat model of the human colonic microflora.

PubMed

Carman, Robert J; Simon, Mary Alice; Petzold, H Earl; Wimmer, Robert F; Batra, Monica R; Fernández, A Haydée; Miller, Margaret A; Bartholomew, Mary

2005-11-01

A chemostat model of the healthy human large bowel ecosystem was used to establish no effect levels for tetracycline, neomycin, and erythromycin. For each compound, the equivalent to four oral doses (0, 1.5, 15, and 150 mg/60 kg person/d) was studied. Concentrations of the test compounds in the chemostat medium were intended to simulate fecal levels that might be expected following consumption of food containing antibiotic residue and were based on published oral doses and fecal levels. We monitored the following parameters: short chain fatty acids, bile acids, sulfate reduction, azoreductase and nitroreductase activities, beta-glucosidase and beta-glucuronidase activities, a range of bacterial counts and, lastly, the susceptibility among sentinel bacteria to each test compound. Neomycin and erythromycin reduced bile acid metabolism. Neomycin elevated propionate levels and caused a marginal diminution in azoreductase activity. Based on our results, the no observed effect level (NOEL) of both tetracycline and erythromycin was 15 mg/60 kg person/d. The NOEL for neomycin was 1.5 mg/60 kg person/d.

Competitive-level differences in Yo-Yo intermittent recovery and twelve minute run test performance in soccer referees.

PubMed

Castagna, Carlo; Abt, Grant; D'Ottavio, Stefano

2005-11-01

The aim of this study was to examine yo-yo intermittent recovery test (Yo-Yo test) and 12-minute run test (12MRT) performances in experienced soccer referees of different competitive levels. Three groups (n = 14 each) of experienced Italian soccer referees officiating in the first (series AB, top-level), third (series C, medium-level), and fourth (series D, low-level) division, were randomly submitted to the 12MRT and the Yo-Yo test during 2 testing sessions, 48-hours apart. 12MRT performances were 3,000 +/- 112 m; 2,894 +/- 99 m; and 2,896 +/- 171 m for top-level, medium-level and low-level referees, respectively (p > 0.05). In the Yo-Yo test, the top-level, medium-level, and low-level referees covered 1,874 +/- 431 m; 1,360 +/- 172 m; and 1,272 +/- 215 m, respectively. The test performances of top-level referees in the Yo-Yo test was significantly different from those scored by medium-level and low-level referees (p < 0.05). After the Yo-Yo test, blood lactate concentrations (BLC) were higher in the medium-level and low-level referees compared with the top-level referees (p < 0.05). The results of the present study show that the Yo-Yo test and not the 12MRT can discriminate endurance performance in experienced elite level soccer referees. With respect to its discriminative and match performance validity, the Yo-Yo test may be considered a relevant field test to assess endurance preparedness for experienced soccer referees and a useful tool in talent selection.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shamoo, D.A.; Johnson, T.R.; Trim, S.C.

We investigated summer activity patterns in a panel of volunteers drawn from a population segment with potentially high exposure to ambient oxidant pollution. The subjects were 15 men and 5 women aged 19-50, all of whom worked outdoors in the Los Angeles area at least 10 hr per week. The general approach was to (i) calibrate the relationship between ventilation rate (VR) and heart rate (HR) for each subject in controlled exercise; (ii) have subjects monitor their own normal activities with diaries and electronic HR recorders; (iii) estimate VR from HR recordings; and (iv) relate VR with diary descriptions ofmore » activities. Calibration data were fit to the equation log (VR) = (intercept) + (slope x HR), intercept and slope being determined separately for each individual to provide a specific equation to predict her/his VR from measured HR. Individuals' correlation coefficients relating log (VR) with HR ranged from 0.83 to 0.95. Subjects monitored themselves for three 24-hr periods during one week, including their most active work day and their most active non-work day. They wore Heart Watch(R) athletic training instruments which recorded HR once per minute; and recorded each change in their activity, location, or breathing rate in diaries. Breathing rates were classified as sleep, slow, medium, or fast. Diaries showed that sleep occupied about 33% of subject's time, slow activity 59%, medium 7%, and fast 1%. Fast activity was reported only at leisure, never at work. For the group, arithmetic means and standard deviations of predicted VR were 7 +/- 3 L/min for sleep, 12 +/- 7 for slow activity, 14 +/- 8 for medium, and 44 +/- 36 for fast. For the group and for most individuals, distributions of predicted VR within a given activity level (breathing rate) were approximately lognormal, with many values in a narrow range below the arithmetic mean and fewer values in a broader range above it.« less

Mean Cancer Mortality Rates in Low Versus High Elevation Counties in Texas

PubMed Central

Hart, John

2010-01-01

There is controversy as to whether low levels of radiation (i.e., < 5 rem) pose a health risk. This brief inquiry compares archived cancer mortality data in counties having relatively low (0–250 feet above sea level), medium (500–1000 feet above sea level), and high (3000+ feet above sea level) elevations also having corresponding greater natural background levels of radiation respectively. Cancer mortality was found to be lowest in the high elevation counties (mean = 58.2) followed by low elevation counties (67.5) and then medium elevation counties (70.4). Statistically significant differences were found between low –high elevations (p = 0.003), and medium – high elevations (p = 0.010), but not between low and medium elevations (p = 0.5). More rigorous research, with an accounting of confounding variables, is indicated. PMID:21191484

High-level expression and efficient purification of bioactive swollenin in Aspergillus oryzae.

PubMed

Wang, Meihua; Cai, Jin; Huang, Lei; Lv, Zhengbin; Zhang, Yaozhou; Xu, Zhinan

2010-11-01

The bioactivity of swollenin is beneficial to cellulose decomposition by cellulase despite the lack of hydrolytic activity itself. In order to improve the productivity of swollenin, the effects of culture conditions on the expression level in recombinant Aspergillus oryzae were investigated systematically. With regard to the bioactivity of swollenin, glycerin and peanut meal were the optimal carbon or nitrogen source, respectively. The highest level production of swollenin (50 mg L(-1)) was attained after 88 h cultivation with the initial pH of 5.6 in the culture medium. Then the soluble swollenin was effectively purified from the cultural supernatant by ammonium sulfate precipitation and cationic exchange chromatography with recovery yield of 53.2%. The purified swollenin was fully bioactive due to its strong synergistic activity with cellulose.

The antimicrobial activity of heterotrophic bacteria isolated from the marine sponge Erylus deficiens (Astrophorida, Geodiidae)

PubMed Central

Graça, Ana Patrícia; Viana, Flávia; Bondoso, Joana; Correia, Maria Inês; Gomes, Luis; Humanes, Madalena; Reis, Alberto; Xavier, Joana R.; Gaspar, Helena; Lage, Olga M.

2015-01-01

Interest in the study of marine sponges and their associated microbiome has increased both for ecological reasons and for their great biotechnological potential. In this work, heterotrophic bacteria associated with three specimens of the marine sponge Erylus deficiens, were isolated in pure culture, phylogenetically identified and screened for antimicrobial activity. The isolation of bacteria after an enrichment treatment in heterotrophic medium revealed diversity in bacterial composition with only Pseudoalteromonas being shared by two specimens. Of the 83 selected isolates, 58% belong to Proteobacteria, 23% to Actinobacteria and 19% to Firmicutes. Diffusion agar assays for bioactivity screening against four bacterial strains and one yeast, revealed that a high number of the isolated bacteria (68.7%) were active, particularly against Candida albicans and Vibrio anguillarum. Pseudoalteromonas, Microbacterium, and Proteus were the most bioactive genera. After this preliminary screening, the bioactive strains were further evaluated in liquid assays against C. albicans, Bacillus subtilis and Escherichia coli. Filtered culture medium and acetone extracts from three and 5 days-old cultures were assayed. High antifungal activity against C. albicans in both aqueous and acetone extracts as well as absence of activity against B. subtilis were confirmed. Higher levels of activity were obtained with the aqueous extracts when compared to the acetone extracts and differences were also observed between the 3 and 5 day-old extracts. Furthermore, a low number of active strains was observed against E. coli. Potential presence of type-I polyketide synthases (PKS-I) and non-ribosomal peptide synthetases (NRPSs) genes were detected in 17 and 30 isolates, respectively. The high levels of bioactivity and the likely presence of associated genes suggest that Erylus deficiens bacteria are potential sources of novel marine bioactive compounds. PMID:25999928

Fluorescence-Based Sensor for Monitoring Activation of Lunar Dust

NASA Technical Reports Server (NTRS)

Wallace, William T.; Jeevarajan, Antony S.

2012-01-01

This sensor unit is designed to determine the level of activation of lunar dust or simulant particles using a fluorescent technique. Activation of the surface of a lunar soil sample (for instance, through grinding) should produce a freshly fractured surface. When these reactive surfaces interact with oxygen and water, they produce hydroxyl radicals. These radicals will react with a terephthalate diluted in the aqueous medium to form 2-hydroxyterephthalate. The fluorescence produced by 2-hydroxyterephthalate provides qualitative proof of the activation of the sample. Using a calibration curve produced by synthesized 2-hydroxyterephthalate, the amount of hydroxyl radicals produced as a function of sample concentration can also be determined.

Two Distinct States of Escherichia coli Cells That Overexpress Recombinant Heterogeneous β-Galactosidase*

PubMed Central

Zhao, Yun; He, Wei; Liu, Wei-Feng; Liu, Chun-Chun; Feng, Li-Kui; Sun, Lei; Yan, Yong-Bin; Hang, Hai-Ying

2012-01-01

The mechanism by which inclusion bodies form is still not well understood, partly because the dynamic processes of the inclusion body formation and its solubilization have hardly been investigated at an individual cell level, and so the important detailed information has not been acquired for the mechanism. In this study, we investigated the in vivo folding and aggregation of Aspergillus phoenicis β-d-galactosidase fused to a red fluorescence protein in individual Escherichia coli cells. The folding status and expression level of the recombinant β-d-galactosidase at an individual cell level was analyzed by flow cytometry in combination with transmission electron microscopy and Western blotting. We found that individual E. coli cells fell into two distinct states, one containing only inclusion bodies accompanied with low galactosidase activity and the other containing the recombinant soluble galactosidase accompanied with high galactosidase activity. The majority of the E. coli cells in the later state possessed no inclusion bodies. The two states of the cells were shifted to a cell state with high enzyme activity by culturing the cells in isopropyl 1-thio-β-d-galactopyranoside-free medium after an initial protein expression induction in isopropyl 1-thio-β-d-galactopyranoside-containing medium. This shift of the cell population status took place without the level change of the β-d-galactosidase protein in individual cells, indicating that the factor(s) besides the crowdedness of the recombinant protein play a major role in the cell state transition. These results shed new light on the mechanism of inclusion body formation and will facilitate the development of new strategies in improving recombinant protein quality. PMID:22303013

Maintenance of drug metabolism and transport functions in human precision-cut liver slices during prolonged incubation for 5 days.

PubMed

Starokozhko, Viktoriia; Vatakuti, Suresh; Schievink, Bauke; Merema, Marjolijn T; Asplund, Annika; Synnergren, Jane; Aspegren, Anders; Groothuis, Geny M M

2017-05-01

Human precision-cut liver slices (hPCLS) are a valuable ex vivo model that can be used in acute toxicity studies. However, a rapid decline in metabolic enzyme activity limits their use in studies that require a prolonged xenobiotic exposure. The aim of the study was to extend the viability and function of hPCLS to 5 days of incubation. hPCLS were incubated in two media developed for long-term culture of hepatocytes, RegeneMed ® , and Cellartis ® , and in the standard medium WME. Maintenance of phase I and II metabolism was studied both on gene expression as well as functional level using a mixture of CYP isoform-specific substrates. Albumin synthesis, morphological integrity, and glycogen storage was assessed, and gene expression was studied by transcriptomic analysis using microarrays with a focus on genes involved in drug metabolism, transport and toxicity. The data show that hPCLS retain their viability and functionality during 5 days of incubation in Cellartis ® medium. Albumin synthesis as well as the activity and gene expression of phase I and II metabolic enzymes did not decline during 120-h incubation in Cellartis ® medium, with CYP2C9 activity as the only exception. Glycogen storage and morphological integrity were maintained. Moreover, gene expression changes in hPCLS during incubation were limited and mostly related to cytoskeleton remodeling, fibrosis, and moderate oxidative stress. The expression of genes involved in drug transport, which is an important factor in determining the intracellular xenobiotic exposure, was also unchanged. Therefore, we conclude that hPCLS cultured in Cellartis ® medium are a valuable human ex vivo model for toxicological and pharmacological studies that require prolonged xenobiotic exposure.

Enhanced Production of a Recombinant Multidomain Thermostable GH9 Processive Endo-1,4-β-Glucanase (CenC) from Ruminiclostridium thermocellum in a Mesophilic Host Through Various Cultivation and Induction Strategies.

PubMed

Haq, Ikram Ul; Akram, Fatima

2017-09-01

Commonly, unintentional induction and inadvertently preparing medium for engineered Escherichia coli BL21 CodonPlus (DE3)-RIPL, give poor or variable yields of heterologous proteins. Therefore, to enhance the activity and production of an industrially relevant recombinant processive endo-1,4-β-glucanase (CenC) propagated in Escherichia coli BL21 CodonPlus(DE3)-RIPL through various cultivation and induction strategies. Investigation of various growth media and induction parameters revealed that high-cell-density and optimal CenC expression were obtained in ZYBM9 medium induced either with 0.5 mM IPTG/150 mM lactose, after 6 h induction at 37 °C; and before induction, bacterial cells were given heat shock (42 °C) for 1 h when culture density (OD 600nm ) reached at 0.6. Intracellular enzyme activity was enhanced by 6.67 and 3.20-fold in ZYBM9 and 3×ZYBM9 medium, respectively, under optimal conditions. Using YNG auto-induction medium, activity was 2.5-fold increased after 10 h incubation at 37 °C. Approximately similar results were obtained by transferring the optimized process at the bioreactor level. Results showed that the effective process strategy is essential to enhance recombinant bacterial cell mass and enzyme production from small to large-scale. To the best of our knowledge, this is the first ever report on enhanced production of thermostable processive endo-1,4-β-glucanase cloned from Ruminiclostridium thermocellum, which is a suitable candidate for industrial applications. Graphical Abstract Flow Chart Summary of Enhanced Production of a Recombinant Multidomain Thermostable GH9 Processive Endo-1,4-β-glucanase from Ruminiclostridium thermocellum.

Lipolysis of natural long chain and synthetic medium chain galactolipids by pancreatic lipase-related protein 2.

PubMed

Amara, Sawsan; Barouh, Nathalie; Lecomte, Jérôme; Lafont, Dominique; Robert, Sylvie; Villeneuve, Pierre; De Caro, Alain; Carrière, Frédéric

2010-04-01

Monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG) are the most abundant lipids in nature, mainly as important components of plant leaves and chloroplast membranes. Pancreatic lipase-related protein 2 (PLRP2) was previously found to express galactolipase activity, and it is assumed to be the main enzyme involved in the digestion of these common vegetable lipids in the gastrointestinal tract. Most of the previous in vitro studies were however performed with medium chain synthetic galactolipids as substrates. It was shown here that recombinant guinea pig (Cavia porcellus) as well as human PLRP2 hydrolyzed at high rates natural DGDG and MGDG extracted from spinach leaves. Their specific activities were estimated by combining the pH-stat technique, thin layer chromatography coupled to scanning densitometry and gas chromatography. The optimum assay conditions for hydrolysis of these natural long chain galactolipids were investigated and the optimum bile salt to substrate ratio was found to be different from that established with synthetic medium chains MGDG and DGDG. Nevertheless the length of acyl chains and the nature of the galactosyl polar head of the galactolipid did not have major effects on the specific activities of PLRP2, which were found to be very high on both medium chain [1786+/-100 to 5420+/-85U/mg] and long chain [1756+/-208 to 4167+/-167U/mg] galactolipids. Fatty acid composition analysis of natural MGDG, DGDG and their lipolysis products revealed that PLRP2 only hydrolyzed one ester bond at the sn-1 position of galactolipids. PLRP2 might be used to produce lipid and free fatty acid fractions enriched in either 16:3 n-3 or 18:3 n-3 fatty acids, both found at high levels in galactolipids. 2010 Elsevier B.V. All rights reserved.

Energy and glucose pathways in thiamine deficient primary rat brain microvascular endothelial cells.

PubMed

Ham, D; Karska-Wysocki, B

2005-12-01

Thiamine deficiency (TD) results in lactate acidosis, which is associated with neurodegeneration. The aim of this study was to investigate this alteration in primary rat brain endothelia. Spectrophotometric analysis of culture media revealed that only a higher concentration of pyrithiamine, which accelerates the intracellular blocking of thiamine, significantly elevated the lactate level and lactate dehydrogenase activity within 7 days. The medium without pyrithiamine and with a thiamine concentration comparable to pathophysiological plasma levels mildly reduced only the activity of transketolase. This suggests that significant metabolic changes may not occur at the early phase of TD in cerebral capillary cells, while anaerobic glycolysis in capillaries may be mediated during late stage/chronic TD.

Ammonium nitrate and iron nutrition effects on some nitrogen assimilation enzymes and metabolites in Spirulina platensis.

PubMed

Esen, Merve; Ozturk Urek, Raziye

2015-01-01

The effect of various concentrations of ammonium nitrate (5-60 mM), an economical nitrogen source, on the growth, nitrate-ammonium uptake rates, production of some pigments and metabolites, and some nitrogen assimilation enzymes such as nitrate reductase (NR), nitrite reductase (NiR), glutamine synthetase (GS), and glutamate synthase (GOGAT) in Spirulina platensis (Gamont) Geitler was investigated. Ten millimolars of ammonium nitrate stimulated the growth, production of pigments and the other metabolites, and enzyme activities, whereas 30 and 60 mM ammonium nitrate caused inhibition. In the presence of 10 mM ammonium nitrate, different concentrations of iron were tried in the growth media of S. platensis. After achieving the best growth, levels of metabolite and pigment production, and enzyme activities in the presence of 10 mM ammonium nitrate as a nitrogen source, different iron concentrations (10-100 µM) were tried in the growth medium of S. platensis. The highest growth, pigment and metabolite levels, and enzyme activities were determined in the medium containing 50 µM iron and 10 mM ammonium nitrate. In this optimum condition, the highest dry biomass level, chlorophyll a, and pyruvate contents were obtained as 55.42 ± 3.8 mg mL(-1) , 93.114 ± 7.9 µg g(-1) , and 212.5 ± 18.7 µg g(-1) , respectively. The highest NR, NiR, GS, and GOGAT activities were 67.16 ± 5.1, 777.92 ± 52, 0.141 ± 0.01, and 44.45 ± 3.6, respectively. Additionally, 10 mM ammonium nitrate is an economical and efficient nitrogen source for nitrogen assimilation of S. platensis, and 50 µM iron is optimum for the growth of S. platensis. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

Effect of environmental and cultural conditions on medium pH and explant growth performance of Douglas-fir ( Pseudotsuga menziesii) shoot cultures

PubMed Central

Chen, Chien-Chih; Bates, Rick; Carlson, John

2015-01-01

The medium pH level of plant tissue cultures has been shown to be essential to many aspects of explant development and growth. Sensitivity or tolerance of medium pH change in vitro varies according to specific requirements of individual species. The objectives of this study are to 1) determine medium pH change over time in storage conditions and with presence of explants, 2) evaluate the effects of medium pH change on explant growth performance and 3) assess the effects of adding a pH stabilizer, 2-(N-morpholino)ethanesulfonic acid (MES) that is commonly used in Douglas-fir micropropagation medium. Vegetative buds were collected in the spring before breaking dormancy from juvenile and mature donor trees for conducting these evaluations. Medium, with or without MES, was pre-adjusted to five pH levels before adding MES, agar and autoclaving. Medium pH changes and explant growth parameters were measured at eight different incubation times. Overall, MES provided a more stable medium pH, relative to starting pH values, under both light and dark storage conditions as well as with presence of explants. A general trend of decreasing medium pH over time was found comparing explants from juvenile and mature donor genotypes. Explant height and weight growth increased over time, but differ among explants from juvenile and mature donor genotypes. Our findings suggest that a 21-day subculture practice may best sustain medium freshness, medium pH level and desirable explant growth. PMID:26535110

Simultaneous production of both high molecular weight pullulan and oligosaccharides by Aureobasdium melanogenum P16 isolated from a mangrove ecosystem.

PubMed

Liu, Nan-Nan; Chi, Zhe; Wang, Qin-Qing; Hong, Jiang; Liu, Guang-Lei; Hu, Zhong; Chi, Zhen-Ming

2017-09-01

After the compositional change of a pullulan production medium, a molecular weight (Mw) of the pullulan produced by Aureobasidium melanogenum P16 was 2.32×10 6 and a pullulan titer was 44.4g/L while a Mw of the pullulan produced by A. melanogenum P16 grown in the initial medium was only 3.47×10 5 and a pullulan titer was 65.3g/L. The increased Mw of the pullulan was due to the decreased activities of α-amylase, glucoamylase and pullulanase while the decreased pullulan titer was related to the decreased transcriptional levels of the genes encoding 6-P-glucose kinase, glucosyltransferase, α-phosphoglucose mutase, UDPG-pyrophosphorylase and pullulan synthetase. During the 10-L fermentation, when the yeast strain P16 was grown in the initial medium, the pullulan and oligosaccharide titers were 65.5g/L and 7.8g/L, respectively and the Mw of the produced pullulan was 4.42×10 5 while when the yeast strain P16 was grown in the compositionally changed medium, the pullulan and oligosaccharide titers were 46.4g/L and 27.8g/L, respectively and the Mw of the produced pullulan was 2.6×10 6 . Most of the oligosaccharides produced by the yeast strain P16 cultivated in the compositionally changed medium had degree of polymerization of 4 and 5. Therefore, both of the high Mw pullulan and oligosaccharides with high levels were produced by the yeast strain P16. Copyright © 2017 Elsevier B.V. All rights reserved.

Properties of Streptococcus mutans Grown in a Synthetic Medium: Binding of Glucosyltransferase and In Vitro Adherence, and Binding of Dextran/Glucan and Glycoprotein and Agglutination

PubMed Central

Wu-Yuan, Christine D.; Tai, Stella; Slade, Hutton D.

1979-01-01

The influence of culture media on various properties of Streptococcus mutans was investigated. Strains of S. mutans (serotypes c, d, f, and g) were grown in a complex medium (Todd-Hewitt broth [THB]) or a synthetic medium (SYN). The SYN cells, in contrast to THB cells, did not bind extracellular glucosyltransferase and did not produce in vitro adherence. Both types of cells possessed constitutive levels of glucosyltransferase. B13 cells grown in SYN plus invertase-treated glucose possessed the same level of constitutive enzyme as THB cells. In contrast to THB cells, the SYN cells of seven serotype strains did not agglutinate upon the addition of high-molecular-weight dextran/glucan. Significant quantities of lower-molecular-weight (2 × 104 or 7 × 104) dextran and B13 glucan were bound by SYN cells. SYN cells agglutinated weakly in anti-glucan serum (titers, 0 to 16), whereas THB cells possessed titers of 32 to 256. Evidence for the existence of a second binding site in agglutination which does not possess a glucan-like polymer has been obtained. B13 cells grown in invertase-treated THB agglutinated to the same degree as normal THB cells. The nature of this site is unknown. SYN cells possess the type-specific polysaccharide antigen. B13 cells did not bind from THB a glycoprotein which reacts with antisera to the A, B, or T blood group antigens or which allows agglutination upon the addition of dextran. The results demonstrate that S. mutans grown in a chemically defined medium possesse markedly different biochemical and biological activities than cells grown in a complex organic medium. PMID:457252

Acoustic emission feedback control for control of boiling in a microwave oven

DOEpatents

White, Terry L.

1991-01-01

An acoustic emission based feedback system for controlling the boiling level of a liquid medium in a microwave oven is provided. The acoustic emissions from the medium correlated with surface boiling is used to generate a feedback control signal proportional to the level of boiling of the medium. This signal is applied to a power controller to automatically and continuoulsly vary the power applied to the oven to control the boiling at a selected level.

Rappaport-Vassiliadis medium for recovery of Salmonella spp. from low microbial load foods: collaborative study.

PubMed

Hammack, T S; Amaguaña, R M; Andrews, W H; Lerner, I

2001-01-01

Twenty-three laboratories participated in a collaborative study to compare the relative effectiveness of Rappaport-Vassiliadis (RV) medium incubated at 42 degrees C, selenite cystine (SC) broth (35 degrees C), and tetrathionate (TT) broth (35 and 43 degrees C) for recovery of Salmonella from the following foods with a low microbial load: dried egg yolk, dry active yeast, ground black pepper, guar gum, and instant nonfat dry milk. For dry active yeast, lauryl tryptose (LT) broth, incubated at 35 degrees C, was used instead of SC broth. All of the foods were artificially inoculated with single Salmonella serovars, that had been lyophilized before inoculation, at high and low target levels of 0.4 and 0.04 colony forming units/g food, respectively. For analysis of 870 test portions, representing all of the foods except yeast, 249 Salmonella-positive test portions were detected by RV medium, 265 by TT broth (43 degrees C), 268 by TT broth (35 degrees C), and 269 by SC broth (35 degrees C). For analysis of 225 test portions of yeast, 79 Salmonella-positive test portions were detected by RV medium, 79 by TT broth (43 degrees C), 84 by TT broth (35 degrees C), and 68 by LT broth (35 degrees C). RV medium was comparable to, or even more effective than, the other selective enrichments for recovery of Salmonella from all of the foods except guar gum. It is recommended that RV (42 degrees C) and TT (35 degrees C) be used with foods that have a low microbial load, except for guar gum for which SC (35 degrees C) and TT (35 degrees C) are recommended.

Analysis of work ability and work-related physical activity of employees in a medium-sized business.

PubMed

Wilke, Christiane; Ashton, Philip; Elis, Tobias; Biallas, Bianca; Froböse, Ingo

2015-12-18

Work-related physical activity (PA) and work ability are of growing importance in modern working society. There is evidence for age- and job-related differences regarding PA and work ability. This study analyses work ability and work-related PA of employees in a medium-sized business regarding age and occupation. The total sample consists of 148 employees (116 men-78.38% of the sample-and 32 women, accounting for 21.62%; mean age: 40.85 ± 10.07 years). 100 subjects (67.57%) are white-collar workers (WC), and 48 (32.43%) are blue-collar workers (BC). Work ability is measured using the work ability index, and physical activity is obtained via the Global Physical Activity Questionnaire. Work ability shows significant differences regarding occupation (p = 0.001) but not regarding age. Further, significant differences are found for work-related PA concerning occupation (p < 0.0001), but again not for age. Overall, more than half of all subjects meet the current guidelines for physical activity. Work ability is rated as good, yet, a special focus should lie on the promotion during early and late working life. Also, there is still a lack of evidence on the level of work-related PA. Considering work-related PA could add to meeting current activity recommendations.

The effect of income and occupation on body mass index among women in the Cebu Longitudinal Health and Nutrition Surveys (1983-2002).

PubMed

Colchero, M Arantxa; Caballero, Benjamin; Bishai, David

2008-05-01

We assessed the effects of changes in income and occupational activities on changes in body weight among 2952 non-pregnant women enrolled in the Cebu Longitudinal Health and Nutrition Surveys between 1983 and 2002. On average, body mass index (BMI) among women occupied in low activities was 0.29 kg/m(2) (standard error 0.11) larger compared to women occupied in heavy activities. BMI among women involved in medium activities was on average 0.12 kg/m(2) (standard error 0.05) larger compared to women occupied in heavy activities. A one-unit increase in log household income in the previous survey was associated with a small and positive change in BMI of 0.006 kg/m(2) (standard error 0.02) but the effect was not significant. The trend of increasing body mass was higher in the late 1980s than during the 1990s. These period effects were stronger for the women who were younger at baseline and for women with low or medium activity levels. Our analysis suggests a trend in the environment over the last 20 years that has increased the susceptibility of Filipino women to larger body mass.

Phenotypic Restoration by Molybdate of Nitrate Reductase Activity in chlD Mutants of Escherichia coli

PubMed Central

Glaser, J. H.; DeMoss, J. A.

1971-01-01

ChlD mutants of Escherichia coli are pleiotropic, lacking formate-nitrate reductase activity as well as formate-hydrogenlyase activity. Whole-chain formate-nitrate reductase activity, assayed with formate as the electron donor and measuring the amount of nitrite produced, was restored to wild-type levels in the mutants by addition of 10−4m molybdate to the growth medium. Under these conditions, the activity of each of the components of the membrane-bound nitrate reductase chain increased after molybdate supplementation. In the absence of nitrate, the activities of the formate-hydrogenlyase system were also restored by molybdate. Strains deleted for the chlD gene responded in a similar way to molybdate supplementation. The concentration of molybdenum in the chlD mutant cells did not differ significantly from that in the wild-type cells at either low or high concentrations of molybdate in the medium. However, the distribution of molybdenum between the soluble protein and membrane fractions differed significantly from wild type. We conclude that the chlD gene product cannot be a structural component of the formate-hydrogenlyase pathway or the formate-nitrate reductase pathway, but that it must have an indirect role in processing molybdate to a form necessary for both electron transport systems. PMID:4942767

Mapping eco-environmental vulnerability patterns: An assessment framework based on remote sensing, GIS, and AHP

NASA Astrophysics Data System (ADS)

Anh, N. K.; Liou, Y. A.; Li, M. H.

2016-12-01

The motivation for this study is assessment of the eco-environment vulnerability based on four independent determinants: hydro-meteorology, topography, land resources, and human activities. An assessment framework is proposed to assess the vulnerable eco-environment by using 16 variables with 6 of them constructed from Landsat 8 satellite images. The remaining variables were extracted from digital maps. Each variable was evaluated and spatially mapped with the aid of an analytical hierarchy process (AHP) and geographical information system (GIS). The Thua Thien - Hue Province that has been experiencing natural disasters and urbanization in the recent decades is selected as our study area. An eco-environmental vulnerability map is assorted into six vulnerable levels consisting of potential, slight, light, medium, heavy, and very heavy vulnerabilities, representing 14%, 27%, 17%, 26%, 13%, 3% of the study area, respectively. It is found that heavy and very heavy vulnerable areas appear mainly in the low and medium lands with high intensification of social-economic activities and often suffer from flooding. Tiny percentages of medium and heavy vulnerable levels occur in high land areas probably caused by agricultural practices in highlands, slash and burn cultivation and removal of natural forests with new plantation forests and these regions are usually influenced by landslides, flash flooding. Based on our results, three ecological zones requiring different development and protection solutions are proposed to restore local eco-environment toward sustainable development. Our findings support the idea that eco-environmental vulnerability is driven by anthropogenic processes and enhanced by natural disaster in the Thua Thien-Hue Province.

Rapid activation of gill Na+,K+-ATPase in the euryhaline teleost Fundulus heteroclitus

USGS Publications Warehouse

Mancera, J.M.; McCormick, S.D.

2000-01-01

The rapid activation of gill Na+,K+-ATPase was analyzed in the mummichog (Fundulus heteroclitus) and Atlantic salmon (Salmo salar) transferred from low salinity (0.1 ppt) to high salinity (25-35 ppt). In parr and presmolt, Salmo salar gill Na+,K+-ATPase activity started to increase 3 days after transfer. Exposure of Fundulus heteroclitus to 35 ppt seawater (SW) induced a rise in gill Na+,K+-ATPase activity 3 hr after transfer. After 12 hr, the values dropped to initial levels but showed a second significant increase 3 days after transfer. The absence of detergent in the enzyme assay resulted in lower values of gill Na+,K+-ATPase, and the rapid increase after transfer to SW was not observed. Na+,K+-ATPase activity of gill filaments in vitro for 3 hr increased proportionally to the osmolality of the culture medium (600 mosm/kg > 500 mosm/kg > 300 mosm/kg). Osmolality of 800 mosm/kg resulted in lower gill Na+,K+-ATPase activity relative to 600 mosm/kg. Increasing medium osmolality to 600 mosm/kg with mannitol also increased gill Na+,K+-ATPase. Cycloheximide inhibited the increase in gill Na+,K+-ATPase activity observed in hyperosmotic medium in a dose-dependent manner (10-4 M > 10-5 M > 10-6 M). Actinomycin D or bumetanide in the culture (doses of 10-4 M, 10-5 M, and 10-6 M) did not affect gill Na+,K+-ATPase. Injection of fish with actinomycin D prior to gill organ culture, however, prevented the increase in gill Na+,K+-ATPase activity in hyperosmotic media. The results show a very rapid and transitory increase in gill Na+,K+-ATPase activity in the first hours after the transfer of Fundulus heteroclitus to SW that is dependent on translational and transcriptional processes. (C) 2000 Wiley-Liss, Inc.

Extraordinary variation of pump light intensity inside a four-level solid-state laser medium

NASA Astrophysics Data System (ADS)

Qin, Hua; Fu, Rulian; Wang, Zhaoqi; Liu, Juan

2008-08-01

A theoretical investigation of the absorption of the pump light at different intensities through a four-level solid-state laser medium is presented. It is found that the variation of the pump intensity inside the laser medium cannot always simply be dominated by Beer's law. Transmission of the pump light through this laser medium is closely related to the pump intensity itself. In fact, when the pump intensity is relatively low, whose values depend on the characteristics of the medium, the variation of the pump light through the laser medium is consistent with Beer's law. However, while the pump intensity is high enough, the relationship between the transmission of the pump light and its propagation distance is demonstrated to be linear. These theoretical results have been confirmed by the experiment with a medium of YAG:Nd.

A Novel Strategy to Increase the Proliferative Potential of Adult Human β-Cells While Maintaining Their Differentiated Phenotype

PubMed Central

Aly, Haytham; Rohatgi, Nidhi; Marshall, Connie A.; Grossenheider, Tiffani C.; Miyoshi, Hiroyuki; Stappenbeck, Thaddeus S.; Matkovich, Scot J.; McDaniel, Michael L.

2013-01-01

Our previous studies demonstrated that Wnt/GSK-3/β-catenin and mTOR signaling are necessary to stimulate proliferative processes in adult human β-cells. Direct inhibition of GSK-3, that engages Wnt signaling downstream of the Wnt receptor, increases β-catenin nuclear translocation and β-cell proliferation but results in lower insulin content. Our current goal was to engage canonical and non-canonical Wnt signaling at the receptor level to significantly increase human β-cell proliferation while maintaining a β-cell phenotype in intact islets. We adopted a system that utilized conditioned medium from L cells that expressed Wnt3a, R-spondin-3 and Noggin (L-WRN conditioned medium). In addition we used a ROCK inhibitor (Y-27632) and SB-431542 (that results in RhoA inhibition) in these cultures. Treatment of intact human islets with L-WRN conditioned medium plus inhibitors significantly increased DNA synthesis ∼6 fold in a rapamycin-sensitive manner. Moreover, this treatment strikingly increased human β-cell proliferation ∼20 fold above glucose alone. Only the combination of L-WRN conditioned medium with RhoA/ROCK inhibitors resulted in substantial proliferation. Transcriptome-wide gene expression profiling demonstrated that L-WRN medium provoked robust changes in several signaling families, including enhanced β-catenin-mediated and β-cell-specific gene expression. This treatment also increased expression of Nr4a2 and Irs2 and resulted in phosphorylation of Akt. Importantly, glucose-stimulated insulin secretion and content were not downregulated by L-WRN medium treatment. Our data demonstrate that engaging Wnt signaling at the receptor level by this method leads to necessary crosstalk between multiple signaling pathways including activation of Akt, mTOR, Wnt/β-catenin, PKA/CREB, and inhibition of RhoA/ROCK that substantially increase human β-cell proliferation while maintaining the β-cell phenotype. PMID:23776620

A novel strategy to increase the proliferative potential of adult human β-cells while maintaining their differentiated phenotype.

PubMed

Aly, Haytham; Rohatgi, Nidhi; Marshall, Connie A; Grossenheider, Tiffani C; Miyoshi, Hiroyuki; Stappenbeck, Thaddeus S; Matkovich, Scot J; McDaniel, Michael L

2013-01-01

Our previous studies demonstrated that Wnt/GSK-3/β-catenin and mTOR signaling are necessary to stimulate proliferative processes in adult human β-cells. Direct inhibition of GSK-3, that engages Wnt signaling downstream of the Wnt receptor, increases β-catenin nuclear translocation and β-cell proliferation but results in lower insulin content. Our current goal was to engage canonical and non-canonical Wnt signaling at the receptor level to significantly increase human β-cell proliferation while maintaining a β-cell phenotype in intact islets. We adopted a system that utilized conditioned medium from L cells that expressed Wnt3a, R-spondin-3 and Noggin (L-WRN conditioned medium). In addition we used a ROCK inhibitor (Y-27632) and SB-431542 (that results in RhoA inhibition) in these cultures. Treatment of intact human islets with L-WRN conditioned medium plus inhibitors significantly increased DNA synthesis ∼6 fold in a rapamycin-sensitive manner. Moreover, this treatment strikingly increased human β-cell proliferation ∼20 fold above glucose alone. Only the combination of L-WRN conditioned medium with RhoA/ROCK inhibitors resulted in substantial proliferation. Transcriptome-wide gene expression profiling demonstrated that L-WRN medium provoked robust changes in several signaling families, including enhanced β-catenin-mediated and β-cell-specific gene expression. This treatment also increased expression of Nr4a2 and Irs2 and resulted in phosphorylation of Akt. Importantly, glucose-stimulated insulin secretion and content were not downregulated by L-WRN medium treatment. Our data demonstrate that engaging Wnt signaling at the receptor level by this method leads to necessary crosstalk between multiple signaling pathways including activation of Akt, mTOR, Wnt/β-catenin, PKA/CREB, and inhibition of RhoA/ROCK that substantially increase human β-cell proliferation while maintaining the β-cell phenotype.

Effects of nutrient deprivation and differentiation on the expression of growth-arrest genes (gas and gadd) in F9 embryonal carcinoma cells.

PubMed Central

Fleming, J V; Hay, S M; Harries, D N; Rees, W D

1998-01-01

The growth-arrest genes (gas and gadd) are widely expressed during mammalian embryogenesis and may be useful as markers of nutritional stress in the embryo. F9 embryonal carcinoma cells have been used to characterize the effect of serum or amino acid deficiency on growth-arrest gene expression in a differentiating embryonic cell. The differentiation markers, homeobox B2 (HoxB2), collagen type IV and laminin B2, were not induced by growth arrest. Treatment with all-trans retinoic acid (RA) produced a dose-dependent increase in alkaline phosphatase activity, which was unchanged in lysine-deficient medium and reduced in low-serum medium. Low-serum medium also reduced HoxB2 expression. There was a transient 2-6-fold increase in mRNAs for C/EBP-beta, gadd153/CHOP-10 and gas5 genes 24 h after transfer to amino-acid-deficient media. The mRNAs for the gas2 and gas6 genes began to rise slowly by 5-10-fold after a delay of approx. 24 h. The transient increases did not occur in low-serum medium where there was a much smaller and slower increase. Differentiation caused 1-2-fold increases in gas2, gas3 and gas6 mRNA levels. The transient overexpression of gas5, gadd153/CHOP-10 and CCAAT-enhancer-binding protein-beta, and the later expression of gas6 mRNAs in response to amino acid deficiency, were not affected by differentiation. RA treatment increased the expression of gas3 and caused gas2 to be transiently overexpressed in amino-acid-deficient medium. Differentiation in serum-deficient medium did not significantly alter the levels of the growth-arrest gene mRNAs. These results show that in F9 cells the growth-arrest genes are expressed sequentially as a result of nutrient stress. PMID:9461558

Effects of nutrient deprivation and differentiation on the expression of growth-arrest genes (gas and gadd) in F9 embryonal carcinoma cells.

PubMed

Fleming, J V; Hay, S M; Harries, D N; Rees, W D

1998-02-15

The growth-arrest genes (gas and gadd) are widely expressed during mammalian embryogenesis and may be useful as markers of nutritional stress in the embryo. F9 embryonal carcinoma cells have been used to characterize the effect of serum or amino acid deficiency on growth-arrest gene expression in a differentiating embryonic cell. The differentiation markers, homeobox B2 (HoxB2), collagen type IV and laminin B2, were not induced by growth arrest. Treatment with all-trans retinoic acid (RA) produced a dose-dependent increase in alkaline phosphatase activity, which was unchanged in lysine-deficient medium and reduced in low-serum medium. Low-serum medium also reduced HoxB2 expression. There was a transient 2-6-fold increase in mRNAs for C/EBP-beta, gadd153/CHOP-10 and gas5 genes 24 h after transfer to amino-acid-deficient media. The mRNAs for the gas2 and gas6 genes began to rise slowly by 5-10-fold after a delay of approx. 24 h. The transient increases did not occur in low-serum medium where there was a much smaller and slower increase. Differentiation caused 1-2-fold increases in gas2, gas3 and gas6 mRNA levels. The transient overexpression of gas5, gadd153/CHOP-10 and CCAAT-enhancer-binding protein-beta, and the later expression of gas6 mRNAs in response to amino acid deficiency, were not affected by differentiation. RA treatment increased the expression of gas3 and caused gas2 to be transiently overexpressed in amino-acid-deficient medium. Differentiation in serum-deficient medium did not significantly alter the levels of the growth-arrest gene mRNAs. These results show that in F9 cells the growth-arrest genes are expressed sequentially as a result of nutrient stress.

Supplementing glycosylation: A review of applying nucleotide-sugar precursors to growth medium to affect therapeutic recombinant protein glycoform distributions.

PubMed

Blondeel, Eric J M; Aucoin, Marc G

2018-06-15

Glycosylation is a critical quality attribute (CQA) of many therapeutic proteins, particularly monoclonal antibodies (mAbs), and is a major consideration in the approval of biosimilar biologics due to its effects to therapeutic efficacy. Glycosylation generates a distribution of glycoforms, resulting in glycoproteins with inherent molecule-to-molecule heterogeneity, capable of activating (or failing to activate) different effector functions of the immune system. Glycoforms can be affected by the supplementation of nucleotide-sugar precursors, and related components, to culture growth medium, affecting the metabolism of glycosylation. These supplementations has been demonstrated to increase nucleotide-sugar intracellular pools, and impact glycoform distributions, but with varied results. These variations can be attributed to five key factors: Differences between cell platforms (enzyme/transporter expression levels); differences between recombinant proteins produced (glycan-site accessibility); the fermentation and sampling timeline (glucose availability and exoglycosidase accumulation); glutamine levels (affecting ammonia levels, which impact Golgi pH, as well as UDP-GlcNAc pools); and finally, a lack of standardized metrics for observing shifts in glycoform distributions (glycosylation indices) across different experiments. The purpose of this review is to provide detail and clarity on the state of the art of supplementation strategies for nucleotide-sugar precursors for affecting glycosylation in cell culture processes, and to apply glycosylation indices for standardized comparisons across the field. Copyright © 2018. Published by Elsevier Inc.

Growth Performance, Mineral Digestibility, and Blood Characteristics of Ostriches Receiving Drinking Water Supplemented with Varying Levels of Chelated Trace Mineral Complex.

PubMed

Seyfori, Hossein; Ghasemi, Hossein Ali; Hajkhodadadi, Iman; Nazaran, Mohammad Hassan; Hafizi, Maryam

2018-05-01

The effects of water supplementation of chelated trace minerals (CTM, which is named Bonzaplex designed with chelate compounds technology) on growth performance, apparent total tract digestibility (ATTD) of minerals, and some blood metabolites, TM, and antioxidant enzyme values in African ostriches were investigated from 8 to 12 months of age. A total of 20 8-month-old ostriches (five birds in five replicate pens) was randomly allocated into one of the following four treatments: (1) control (basal diet + tap water), (2) low CTM (basal diet +100 mg/bird/day CTM powder in tap water), (3) medium CTM (basal diet +1 g/bird/day CTM powder in tap water), and (4) high CTM (basal diet +2 g/bird/day CTM powder in tap water). Compared with control, medium CTM improved (P < 0.05) daily weight gain and ATTD of phosphorous, zinc, and copper in 12-month-old ostriches. Furthermore, the feed conversion ratio was lower, and ATTD of magnesium was higher in the medium- and high-CTM groups than that in the control group (P < 0.05). At the end of the trial, ostriches receiving high-CTM treatment exhibited the lower (P < 0.05) serum triglyceride and very low-density lipoprotein cholesterol concentrations and higher copper levels compared to those of the control treatment. Supplementation of higher amounts of CTM (medium and high CTM) also increased the activity of serum superoxide dismutase (P < 0.05). No differences were detected for other blood parameters including glucose, total protein, albumin, cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, iron, magnesium, and glutathione peroxidase values. In conclusion, supplementation of CTM at the level of 1 g/bird/day to the drinking water can be recommended for improving growth performance, mineral absorption, and antioxidant status of ostriches fed diets containing the recommended levels of inorganic TM.

Comparison of gait after Syme and transtibial amputation in children: factors that may play a role in function.

PubMed

Jeans, Kelly A; Karol, Lori A; Cummings, Donald; Singhal, Kunal

2014-10-01

Preservation of maximal limb length during amputation is often recommended to maximize the efficiency and symmetry of gait. The goals of this study were to determine (1) whether there are gait differences between children with a Syme (or Boyd) amputation and those with a transtibial-level amputation, and (2) whether the type of prosthetic foot affects gait and PODCI (Pediatric Outcomes Data Collection Instrument) outcomes. Sixty-four patients (age range, 4.7 to 19.2 years) with unilateral below-the-knee prosthesis use (forty-one in the Syme group and twenty-three in the transtibial group) underwent gait analysis and review of data for the involved limb. The twelve prosthetic foot types were categorized as designed for a high, medium, or low activity level (e.g., Flex foot, dynamic response foot, or SACH). Statistical analyses were conducted. Kinematic differences of <4° in total prosthetic ankle motion and 8° in external hip rotation were seen between the Syme and transtibial groups. Ankle power was greater in the transtibial group, whereas the Syme group had greater coronal-plane hip power (p < 0.05). Prosthetic ankle motion was significantly greater in the high compared with the medium and low-performance feet. However, the PODCI happiness score was higher in patients with low compared with medium-performance feet (p < 0.05). Small differences in prosthetic ankle motion and power were found between children with Syme and transtibial amputations. Ankle motion was greater in patients using high-performance feet (9% of the total cohort) compared with medium-performance (59%) and low-performance (31%) feet. Despite the increased ankle motion achieved with high-performance dynamic feet, this advantage was not reflected in peak power of the prosthetic ankle or the PODCI sports/physical functioning subscale. Therapeutic Level III. See Instructions for Authors for a complete description of levels of evidence. Copyright © 2014 by The Journal of Bone and Joint Surgery, Incorporated.

Response of Staphylococcus aureus isolates from bovine mastitis to exogenous iron sources.

PubMed

Diarra, M S; Petitclerc, D; Lacasse, P

2002-09-01

Staphylococcus aureus can survive in conditions of extremely low iron concentration. The ability of S. aureus to use two exogenous hydroxamate types of siderophores (desferrioxamine and ferrichrome) and four iron-containing proteins found in cattle (hemin, hemoglobin, ferritin, and lactoferrin) were tested on 16 reference and clinical isolates. For all strains tested, ferrichrome and desferrioxamine showed strong growth-promoting activities in a disk diffusion assay and in liquid medium. The heme proteins hemin and hemoglobin were also found to support growth in culture media lacking other iron sources, while lactoferrin failed to do so. On media containing the iron chelator dipyridyl, ferritin induced a growth inhibition effect that was further enhanced in the presence of lactoferrin in seven of the 13 tested strains. Staphylococcus aureus was able to bind hemin and the level of binding activity was not increased after growth in iron-rich or -poor media. Dot-blot competition tests showed that biotin-labeled lactoferrin binds to S. aureus, and this binding can be inhibited by unlabeled lactoferrin. Expression of lactoferrin-binding activity was independent of the level of iron in the medium and the iron saturation status of lactoferrin. For each strain tested, ligand blots showed lactoferrin-binding proteins of molecular weights ranging from 32 to 92 kDa. Possible functions of these lactoferrin-binding proteins could not be related to iron acquisition mechanism in S. aureus.

Electrochemical sample matrix elimination for trace-level potentiometric detection with polymeric membrane ion-selective electrodes.

PubMed

Chumbimuni-Torres, Karin Y; Calvo-Marzal, Percy; Wang, Joseph; Bakker, Eric

2008-08-01

Potentiometric sensors are today sufficiently well understood and optimized to reach ultratrace level (subnanomolar) detection limits for numerous ions. In many cases of practical relevance, however, a high electrolyte background hampers the attainable detection limits. A particularly difficult sample matrix for potentiometric detection is seawater, where the high saline concentration forms a major interfering background and reduces the activity of most trace metals by complexation. This paper describes for the first time a hyphenated system for the online electrochemically modulated preconcentration and matrix elimination of trace metals, combined with a downstream potentiometric detection with solid contact polymeric membrane ion-selective microelectrodes. Following the preconcentration at the bismuth-coated electrode, the deposited metals are oxidized and released to a medium favorable to potentiometric detection, in this case calcium nitrate. Matrix interferences arising from the saline sample medium are thus circumvented. This concept is successfully evaluated with cadmium as a model trace element and offers potentiometric detection down to low parts per billion levels in samples containing 0.5 M NaCl background electrolyte.

Electrochemical Sample Matrix Elimination for Trace Level Potentiometric Detection with Polymeric Membrane Ion-Selective Electrodes

PubMed Central

Chumbimuni-Torres, Karin Y.; Calvo-Marzal, Percy; Wang, Joseph; Bakker, Eric

2008-01-01

Potentiometric sensors are today sufficiently well understood and optimized to reach ultra-trace level (sub-nanomolar) detection limits for numerous ions. In many cases of practical relevance, however, a high electrolyte background hampers the attainable detection limits. A particularly difficult sample matrix for potentiometric detection is seawater, where the high saline concentration forms a major interfering background and reduces the activity of most trace metals by complexation. This paper describes for the first time a hyphenated system for the online electrochemically modulated preconcentration and matrix elimination (EMPM) of trace metals, combined with a downstream potentiometric detection with solid contact polymeric membrane ion-selective microelectrodes. Following the preconcentration at the bismuth coated electrodes, the deposited metals are oxidized and released to a medium favorable to potentiometric detection, in this case calcium nitrate. Matrix interferences arising from the saline sample medium are thus circumvented. This concept is successfully evaluated with cadmium as a model trace element and offers potentiometric detection down to low parts per billion levels in samples containing 0.5 M NaCl background electrolyte. PMID:18570385

Tretinoin-loaded lipid-core nanocapsules decrease reactive oxygen species levels and improve bovine embryonic development during in vitro oocyte maturation.

PubMed

Lucas, Caroline Gomes; Remião, Mariana Härter; Komninou, Eliza Rossi; Domingues, William Borges; Haas, Cristina; Leon, Priscila Marques Moura de; Campos, Vinicius Farias; Ourique, Aline; Guterres, Silvia S; Pohlmann, Adriana R; Basso, Andrea Cristina; Seixas, Fabiana Kömmling; Beck, Ruy Carlos Ruver; Collares, Tiago

2015-12-01

In vitro oocyte maturation (IVM) protocols can be improved by adding chemical supplements to the culture media. Tretinoin is considered an important retinoid in embryonic development and its association with lipid-core nanocapsules (TTN-LNC) represents an innovative way of improving its solubility, and chemical stability, and reducing its toxicity. The effects of supplementing IVM medium with TTN-LNC was evaluated by analyzing production of reactive oxygen species (ROS), S36-phosphorilated-p66Shc levels and caspase activity in early embryonic development, and expression of apoptosis and pluripotency genes in blastocysts. The lowest concentration tested (0.25μM) of TTN-LNC generated higher blastocyst rate, lower ROS production and S36-p66Shc amount. Additionally, expression of BAX and SHC1 were lower in both non-encapsulated tretinoin (TTN) and TTN-LNC-treated groups. Nanoencapsulation allowed the use of smaller concentrations of tretinoin to supplement IVM medium thus reducing toxic effects related with its use, decreasing ROS levels and apoptose frequency, and improving the blastocyst rates. Copyright © 2015 Elsevier Inc. All rights reserved.

Kinetics of the Active Medium of a Copper Vapor Brightness Amplifier

NASA Astrophysics Data System (ADS)

Kulagin, A. E.; Torgaev, S. N.; Evtushenko, G. S.; Trigub, M. V.

2018-03-01

A spatiotemporal kinetics of the active medium of a copper vapor brightness amplifier is described that allows gain characteristics to be investigated during the pump pulse. Model calculations show that changing the discharge parameters allows the radial gain profiles to be improved significantly, as well as the gain and the inversion duration to be increased. The data obtained will be used to choose the operating conditions for the active medium in the brightness amplifier mode.

Solid-substrate bioprocessing of cow dung for the production of carboxymethyl cellulase by Bacillus halodurans IND18.

PubMed

Vijayaraghavan, P; Prakash Vincent, S G; Dhillon, G S

2016-02-01

The production of carboxymethyl cellulase (CMCase) by Bacillus halodurans IND18 under solid substrate fermentation (SSF) using cow dung was optimized through two level full factorial design and second order response surface methodology (RSM). The central composite design (CCD) was employed to optimize the vital fermentation parameters, such as pH of the substrate, concentration of nitrogen source (peptone) and ion (sodium dihydrogen phosphate) sources in medium for achieving higher enzyme production. The optimum medium composition was found to be 1.46% (w/w) peptone, 0.095% (w/w) sodium dihydrogen phosphate and pH 8.0. The model prediction of 4210IU/g enzyme activity at optimum conditions was verified experimentally as 4140IU/g. The enzyme was active over a broad temperature range (40-60±1°C) and pH (7.0-9.0) with maximal activity at 60±1°C and pH 8.0. This study demonstrated the potential of cow dung as novel substrate for CMCase production. Copyright © 2015 Elsevier Ltd. All rights reserved.

Hepatoprotective Effects of Grape Seed Procyanidin B2 in Rats With Carbon Tetrachloride-induced Hepatic Fibrosis.

PubMed

Wang, Zhenli; Zhang, Zemin; Du, Ning; Wang, Kai; Li, Lei

2015-01-01

Infectious hepatitis is a serious problem affecting millions of people worldwide, particularly in China and other developing countries, and it is the major risk factor for hepatic cirrhosis. To date, the pathogenesis of hepatic cirrhosis is complex and unclear. Traditional Chinese medicine (TCM) has long been used in its treatment; however, little is known to date about the effects of grape seed procyanidin B2 (GSPB2) on liver fibrosis. Using a rat model of carbon tetrachloride (CCl4)-induced hepatic fibrosis, the study intended to investigate the hepatoprotective effects of GSPB2 and to determine the possible pathway by which GSPB2 exerts its activities. Design • Thirty-six male, Sprague-Dawley rats were used in the study. Rats in a model (CCl4 only) group and the GSPB2 group were given CCl4 to induce hepatic fibrosis. Simultaneously, animals in the GSPB2 group were treated with GSPB2 by intragastric administration for 12 wk. In addition, the rat's Kupffer cells were cultured with CCl4 and GSPB2. The study took place at the central laboratory of Qilu Hospital at Shandong University in Jinan, China. The following parameters were investigated: (1) hepatic function; (2) the liver fibrosis index-serum hyaluronic acid (HA), laminin (LN), type 3 procollagen (PC-3), collagen 4, and hepatic hydroxyproline; (3) the expression in the liver of transforming growth factor β-1 (TGF-β1); (4) inflammatory cytokines in the liver and cell culture medium-tumor necrosis factor α (TNF-α), interleukin (IL) 1-β (IL-1β), IL-6, and IL-17; (5) oxidative stress markers in the liver and cell culture medium-malondialdehyde (MDA), 8-hydroxy-2-deoxyguanosine (8-OHdG), total superoxide dismutase (T-SOD), and total antioxidant capacity (T-AOC); and (6) levels of angiotensin 2 (Ang 2) in the liver. The CCl4 induced (1) significant hepatic-function damage; (2) elevated levels of the measures of the liver fibrosis index, TGF-β1, inflammatory cytokines, MDA, and 8-OHdG; (3) a reduction in the activities of T-SOD and T-AOC; and (4) no effect on the level of expression of hepatic Ang 2. GSPB2 treatment partially reversed the changes induced by CCl4. The cell culture also showed that CCl4 elevated the levels of inflammatory cytokines and MDA in the Kupffer cell culture medium, whereas it reduced the activities of T-SOD and T-AOC in the medium. GSPB2 treatment partially reversed the changes induced by CCl4. GSPB2 had hepatoprotective effects on CCl4-induced hepatic fibrosis in Sprague-Dawley rats and inhibited the inflammatory response and oxidative stress in vivo and in vitro.

A novel accelerometer-based method to describe day-to-day exposure to potentially osteogenic vertical impacts in older adults: findings from a multi-cohort study.

PubMed

Hannam, K; Deere, K C; Hartley, A; Clark, E M; Coulson, J; Ireland, A; Moss, C; Edwards, M H; Dennison, E; Gaysin, T; Cooper, R; Wong, A; McPhee, J S; Cooper, C; Kuh, D; Tobias, J H

2017-03-01

This observational study assessed vertical impacts experienced in older adults as part of their day-to-day physical activity using accelerometry and questionnaire data. Population-based older adults experienced very limited high-impact activity. The accelerometry method utilised appeared to be valid based on comparisons between different cohorts and with self-reported activity. We aimed to validate a novel method for evaluating day-to-day higher impact weight-bearing physical activity (PA) in older adults, thought to be important in protecting against osteoporosis, by comparing results between four cohorts varying in age and activity levels, and with self-reported PA levels. Participants were from three population-based cohorts, MRC National Survey of Health and Development (NSHD), Hertfordshire Cohort Study (HCS) and Cohort for Skeletal Health in Bristol and Avon (COSHIBA), and the Master Athlete Cohort (MAC). Y-axis peaks (reflecting the vertical when an individual is upright) from a triaxial accelerometer (sampling frequency 50 Hz, range 0-16 g) worn at the waist for 7 days were classified as low (0.5-1.0 g), medium (1.0-1.5 g) or higher (≥1.5 g) impacts. There were a median of 90, 41 and 39 higher impacts/week in NSHD (age 69.5), COSHIBA (age 76.8) and HCS (age 78.5) participants, respectively (total n = 1512). In contrast, MAC participants (age 68.5) had a median of 14,322 higher impacts/week. In the three population cohorts combined, based on comparison of beta coefficients, moderate-high-impact activities as assessed by PA questionnaire were suggestive of stronger association with higher impacts from accelerometers (0.25 [0.17, 0.34]), compared with medium (0.18 [0.09, 0.27]) and low impacts (0.13 [0.07,0.19]) (beta coefficient, with 95 % CI). Likewise in MAC, reported moderate-high-impact activities showed a stronger association with higher impacts (0.26 [0.14, 0.37]), compared with medium (0.14 [0.05, 0.22]) and low impacts (0.03 [-0.02, 0.08]). Our new accelerometer method appears to provide valid measures of higher vertical impacts in older adults. Results obtained from the three population-based cohorts indicate that older adults generally experience very limited higher impact weight-bearing PA.

Growth and differentiation of a murine interleukin-3-producing myelomonocytic leukemia cell line in a protein-free chemically defined medium.

PubMed

Kajigaya, Y; Ikuta, K; Sasaki, H; Matsuyama, S

1990-10-01

We established the continuous growth of WEHI-3B D+ cells in protein-free chemically defined F-12 medium by stepwise decreases in the concentration of fetal calf serum. This cell line, designated as WEHI-3B-Y1, has now been propagated in protein-free F-12 medium for 3 years. The population-doubling time of the cells in culture is about 24 hr. WEHI-3B-Y1 cells are immature undifferentiated cells which show positive staining for naphthol ASD chloroacetate esterase and alpha-naphthyl butyrate esterase and spontaneously exhibit a low level of differentiation to mature granulocytes and macrophages. Medium conditioned by WEHI-3B-Y1 cells stimulated the proliferation of an interleukin-3 (IL-3)-dependent FDCP-2 cell line. This conditioned medium was shown to have erythroid burst-promoting activity when assayed using normal murine bone marrow. The colony formation of WEHI-3B-Y1 cells in semi-solid agar culture was not stimulated by purified recombinant human granulocyte colony-stimulating factor (rhG-CSF). However, in the presence of human transferrin, rhG-CSF enhanced the number of colonies of WEHI-3B-Y1 cells but did not induce their differentiation. These results suggest that WEHI-3B-Y1 cells cultured in protein-free medium produced murine IL-3. In addition, human G-CSF enhanced the clonal growth but did not induce the differentiation of WEHI-3B-Y1 cells cultured in serum-free medium.

Evolutional schemes for objects with active nuclei

NASA Technical Reports Server (NTRS)

Komberg, B. V.

1979-01-01

The observational properties of quasistellar objects (QSO) reveal that they are extremely violent nuclei of distant galaxies, but the evolutionary stage of these galaxies is still undetermined. Various published attempts to classify QSO under different criteria - including the one based on the morphological type of the surrounding galaxy E- or S- are analyzed. There are evidences that radioactive quasars reside in E-, while radio-quiet quasars reside in both E- and S- systems. The latter may be evolutionary connected to Seyfert-like objects. A correlation between the nuclei activity level in systems of different morphological type and the relative amount of gas in them is noted. From the point of view of activity level and the duration of active stage of nuclei it is concluded that an interaction of galaxies with the intergalactic medium is of particular importance and must be most conspicuous in spheriodal systems of central regions of rich clusters, in tight groups and binary galaxies.

Sodium valproate induces mitochondrial respiration dysfunction in HepG2 in vitro cell model.

PubMed

Komulainen, Tuomas; Lodge, Tiffany; Hinttala, Reetta; Bolszak, Maija; Pietilä, Mika; Koivunen, Peppi; Hakkola, Jukka; Poulton, Joanna; Morten, Karl J; Uusimaa, Johanna

2015-05-04

Sodium valproate (VPA) is a potentially hepatotoxic antiepileptic drug. Risk of VPA-induced hepatotoxicity is increased in patients with mitochondrial diseases and especially in patients with POLG1 gene mutations. We used a HepG2 cell in vitro model to investigate the effect of VPA on mitochondrial activity. Cells were incubated in glucose medium and mitochondrial respiration-inducing medium supplemented with galactose and pyruvate. VPA treatments were carried out at concentrations of 0-2.0mM for 24-72 h. In both media, VPA caused decrease in oxygen consumption rates and mitochondrial membrane potential. VPA exposure led to depleted ATP levels in HepG2 cells incubated in galactose medium suggesting dysfunction in mitochondrial ATP production. In addition, VPA exposure for 72 h increased levels of mitochondrial reactive oxygen species (ROS), but adversely decreased protein levels of mitochondrial superoxide dismutase SOD2, suggesting oxidative stress caused by impaired elimination of mitochondrial ROS and a novel pathomechanism related to VPA toxicity. Increased cell death and decrease in cell number was detected under both metabolic conditions. However, immunoblotting did not show any changes in the protein levels of the catalytic subunit A of mitochondrial DNA polymerase γ, the mitochondrial respiratory chain complexes I, II and IV, ATP synthase, E3 subunit dihydrolipoyl dehydrogenase of pyruvate dehydrogenase, 2-oxoglutarate dehydrogenase and glutathione peroxidase. Our results show that VPA inhibits mitochondrial respiration and leads to mitochondrial dysfunction, oxidative stress and increased cell death, thus suggesting an essential role of mitochondria in VPA-induced hepatotoxicity. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

A strategy to analyse activity-based profiling of tyrosine kinase substrates in OCT-embedded lung cancer tissue.

PubMed

Arni, Stephan; de Wijn, Rik; Garcia-Villegas, Refugio; Bitanihirwe, Byron K Y; Caviezel, Claudio; Weder, Walter; Hillinger, Sven

2018-04-15

The use of optimal cutting temperature (OCT) medium has served to improve the long-term preservation of surgical tissue specimens. Unfortunately, the presence of polymers in OCT has been found to generate signal interference in proteomic-based techniques. Indeed the presence of OCT medium in tissue lysates precludes the analysis of activity based proteomic profiles obtained from lung adenocarcinoma (LuAdCa) resection specimens. In order to probe this question further tissue lysates were prepared from 47 lung non-neoplastic and tumour, node, metastasis (TNM) stage 1 LuAdCa resection specimens embedded with or without OCT, and data of activity based multiplex profiles of protein tyrosine kinase peptide substrates were obtained. We found that changes in overall phosphorylation level coincided with the use of OCT and subsequently developed an OCT per peptide median correcting strategy by performing median centering on the values of each peptide. Application of this post-analytical strategy not only can identify changes in kinase activity but can also assist in identifying novel targets for therapeutic intervention against LuAdCa. Copyright © 2018 Elsevier Inc. All rights reserved.

Algicidal Effects of Prodigiosin on the Harmful Algae Phaeocystis globosa.

PubMed

Zhang, Huajun; Peng, Yun; Zhang, Su; Cai, Guanjing; Li, Yi; Yang, Xujun; Yang, Ke; Chen, Zhangran; Zhang, Jun; Wang, Hui; Zheng, Tianling; Zheng, Wei

2016-01-01

Phaeocystis globosa blooms can have negative effects on higher trophic levels in the marine ecosystem and consequently influence human activities. Strain KA22, identified as the bacterium Hahella, was isolated from coastal surface water and used to control P. globosa growth. A methanol extract from the bacterial cells showed strong algicidal activity. After purification, the compound showed a similar structure to prodigiosin when identified with Q-Exactive Orbitrap MS and nuclear magnetic resonance spectra. The compound showed algicidal activity against P. globosa with a 50% Lethal Dose (LD50) of 2.24 μg/mL. The prodigiosin was stable under heat and acid environment, and it could be degraded under alkaline environment and natural light condition. The growth rates of strain KA22 was fast in 2216E medium and the content of prodigiosin in this medium was more than 70 μg/mL after 16 h incubation. The compound showed particularly strong algicidal activity against Prorocentrum donghaiense, P. Globosa, and Heterosigma akashiwo, but having little effect on three other phytoplankton species tested. The results of our research could increase our knowledge on harmful algal bloom control compound and lead to further study on the mechanisms of the lysis effect on harmful algae.

Antioxidant and antiedema properties of solid-state cultured honey mushroom, Armillaria mellea (higher Basidiomycetes), extracts and their polysaccharide and polyphenol contents.

PubMed

Lai, Min-Nan; Ng, Lean Teik

2013-01-01

Culinary-medicinal honey mushroom or Mi-Huan-Ku, Armillaria mellea (AM), is a popular ingredient in the traditional Chinese medicine for treating diseases of geriatric patients. This study aimed to examine the effect of cultured substrates on the mycelial growth of AM and evaluate its antioxidant and antiedema activities as well as its total polysaccharide and polyphenol contents. Results showed that AM grew best on the maize medium and worst on the potato medium. AM ethanol extract (AM-EtOH) showed stronger DPPH radical scavenging activity than AM aqueous extract (AM-H₂O). However, they were weak in metal chelation and reducing power. AM-EtOH but not AM-H₂O at 200 mg/kg showed antiedema activity in rats. The total β-glucan content of AM-H₂O and AM-EtOH was 21.95% and 3.50%, respectively. AM-EtOH showed higher phenol but lower flavonoid content than AM-H₂O. These results indicate that maize is a good source of substrate for mass production of AM mycelia, and its potency of DPPH radical scavenging and antiedema activities was contributed mainly by the phenolic compounds, not the level of polysaccharide content.

Surface phosphatase in Rhinocladiella aquaspersa: biochemical properties and its involvement with adhesion.

PubMed

Kneipp, Lucimar F; Magalhães, Andressa S; Abi-Chacra, Erika A; Souza, Lucieri O P; Alviano, Celuta S; Santos, André L S; Meyer-Fernandes, José R

2012-08-01

Rhinocladiella aquaspersa is an etiologic agent of chromoblastomycosis, a subcutaneous chronic infectious disease. In the present work, we found that the three morphological forms of this fungus (conidia, mycelia and sclerotic bodies) expressed different levels of ecto-phosphatase activity. Our results demonstrated that surface conidial enzyme is an acid phosphatase, inhibited by sodium salts of molybdate, orthovanadate and fluoride and that the inhibition caused by orthovanadate and molybdate was irreversible. The conidial ecto-phosphatase efficiently released phosphate groups from different phosphorylated substrates, causing a higher rate of phosphate removal when p-nitrophenylphosphate was used as substrate. This ecto-enzyme of R. aquaspersa is modulated by Co(2 +) ions and inorganic phosphate (Pi). Accordingly, removal of Pi from the culture medium resulted in a marked (121-fold) increase of ecto-phosphatase activity. Surface phosphatase activity is apparently involved in fungal adhesive properties, since the attachment of R. aquaspersa to epithelial cells was reversed by the pre-treatment of the conidia with orthovanadate, molybdate and anti-phosphatase antibody. Corroborating this finding, conidia with greater ecto-phosphatase activity (grown in Pi-depleted medium) showed higher adherence to epithelial cells than fungi cultivated in the presence of Pi.

Preservation of nitric oxide-induced relaxation of porcine coronary artery: roles of the dimers of soluble guanylyl cyclase, phosphodiesterase type 5, and cGMP-dependent protein kinase.

PubMed

Liu, Juan; Chen, Zhengju; Ye, Liping; Liu, Huixia; Dou, Dou; Liu, Limei; Yu, Xiaoxing; Gao, Yuansheng

2014-10-01

Soluble guanylyl cyclase (sGC), phosphodiesterase type 5 (PDE5), and guanosine 3',5'-cyclic monophosphate (cGMP)-dependent protein kinase (PKG) are all dimeric. The present study was to determine the role of their dimeric status in nitric oxide-induced vasodilatation. In isolated porcine coronary arteries, after 20 h incubation with serum-free medium, serum-containing medium, or phosphate-buffered saline solution, the protein levels of the dimers of sGC, PDE5, and PKG were diminished while the monomer levels remained unchanged, associated with reduced cGMP elevation in response to DETA NONOate and decreased PDE5 activity; the activity of PKG was not significantly altered. DETA NONOate caused a greater relaxation in arteries incubated for 20 vs. 2 h. The relaxant response was largely abolished by 1H-[1, 2, 4]oxadiazolo[4,3-a]quinoxalin-1-one, an sGC inhibitor. Zaprinast, a PDE5 inhibitor, had no effect on relaxation caused by DETA NONOate of arteries incubated for 20 h but augmented the response incubated for 2 h. A greater relaxation to 8-bromo-guanosine 3'5'-cyclic monophosphate occurred in arteries incubated for 20 than for 2 h. The protein level of the dimers but not monomers of PDE5 was reduced by dithiothreitol and unaffected by hydrogen peroxide, accompanied with decreased PDE5 activity and reduced response to DETA NONOate. These results demonstrate that the dimeric but not monomeric status of sGC and PDE5 of coronary arteries are closely related to their activities. The preserved vasodilator response after 20 h incubation may result in part from a synchronous reduction of the dimer levels of sGC and PDE5 as well as an augmented response to cGMP.

Effect of mono-(2-ethylhexyl) phthalate on steroid production of human granulosa cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Reinsberg, Jochen; Wegener-Toper, Petra; Ven, Katrin van der

2009-08-15

The phthalate ester mono-(2-ethylhexyl) phthalate (MEHP) is the active metabolite of di-(2-ethylhexyl) phthalate, a high-production-volume chemical used as a plasticizer and solvent in numerous consumer products. MEHP has been demonstrated to be a reproductive toxicant in rodents decreasing estradiol and progesterone production in preovulatory granulosa cells. In the present study, we examined the effect of MEHP on steroid production of human granulosa-lutein (GL) cells. Human GL cells collected from women undergoing in vitro fertilization were cultured in medium containing FSH, hCG and 8-Br-cAMP, respectively, together with various concentrations of MEHP (0-500 {mu}mol L{sup -1}). After incubation for 48 h estradiolmore » and progesterone were assayed in the spent culture medium. Furthermore, aromatase activity and mRNA levels of GL cells were determined. Basal as well as FSH-, hCG- and 8-Br-cAMP-stimulated estradiol production of GL cells was suppressed by MEHP in a dose-dependent manner (IC{sub 50} = 105 {mu}mol L{sup -1}, 138 {mu}mol L{sup -1}, 49 {mu}mol L{sup -1} and 78 {mu}mol L{sup -1}). Furthermore aromatase activity and mRNA levels were reduced in GL cells cultured with MEHP. In contrast, MEHP did not alter the production of progesterone up to a concentration of 167 {mu}mol L{sup -1}. The present data indicate that MEHP is a specific inhibitor of estradiol production in human GL cells with a post-cAMP site of action. The inhibition of estradiol production obviously results from a reduction of aromatase activity on the transcript level. As the in vitro effective doses of MEHP are within the range of real environmental exposure levels an inhibitory effect on estrogen production in vivo seems to be possi0009b.« less

Lead-iron phosphate glass as a containment medium for the disposal of high-level nuclear wastes

DOEpatents

Boatner, L.A.; Sales, B.C.

1984-04-11

Disclosed are lead-iron phosphate glasses containing a high level of Fe/sub 2/O/sub 3/ for use as a storage medium for high-level radioactive nuclear waste. By combining lead-iron phosphate glass with various types of simulated high-level nuclear waste

HANFORD MEDIUM-LOW CURIE WASTE PRETREATMENT ALTERNATIVES PROJECT FRACTIONAL CRYSTALLIZATION PILOT SCALE TESTING FINAL REPORT

DOE Office of Scientific and Technical Information (OSTI.GOV)

HERTING DL

2008-09-16

The Fractional Crystallization Pilot Plant was designed and constructed to demonstrate that fractional crystallization is a viable way to separate the high-level and low-activity radioactive waste streams from retrieved Hanford single-shell tank saltcake. The focus of this report is to review the design, construction, and testing details of the fractional crystallization pilot plant not previously disseminated.

Antioxidant Properties of Crude Extract, Partition Extract, and Fermented Medium of Dendrobium sabin Flower

PubMed Central

Abu, Farahziela; Mohd Akhir, Sobri

2017-01-01

Antioxidant properties of crude extract, partition extract, and fermented medium from Dendrobium sabin (DS) flower were investigated. The oven-dried DS flower was extracted using 100% methanol (w/v), 100% ethanol (w/v), and 100% water (w/v). The 100% methanolic crude extract showed the highest total phenolic content (40.33 ± mg GAE/g extract) and the best antioxidant properties as shown by DPPH, ABTS, and FRAP assays. A correlation relationship between antioxidant activity and total phenolic content showed that phenolic compounds were the dominant antioxidant components in this flower extract. The microbial fermentation on DS flower medium showed a potential in increasing the phenolic content and DPPH scavenging activity. The TPC of final fermented medium showed approximately 18% increment, while the DPPH of fermented medium increased significantly to approximately 80% at the end of the fermentation. Dendrobium sabin (DS) flower showed very good potential properties of antioxidant in crude extract and partition extract as well as better antioxidant activity in the flower fermented medium. PMID:28761496

Metabolic activity and behavior of the invasive amphipod Dikerogammarus villosus and two common Central European gammarid species (Gammarus fossarum, Gammarus roeselii): Low metabolic rates may favor the invader.

PubMed

Becker, Jochen; Ortmann, Christian; Wetzel, Markus A; Koop, Jochen H E

2016-01-01

The Ponto-Caspian amphipod Dikerogammarus villosus is one of the most successful invaders in Central European rivers. Contrary to studies on its ecology, ecophysiological studies comparing the species' physiological traits are scarce. In this context, in particular the metabolic activity of the invasive species has rarely been considered and, moreover, the few existing studies on this species report strongly deviating results. The purpose of this study was to assess the metabolic activity and behavior of D. villosus and other common European amphipod species (Gammarus fossarum, Gammarus roeselii) in relation to temperatures covering the thermal regime of the invaded habitats. Based on direct calorimetric measurements of metabolic heat dissipation at three temperature levels (5°C, 15°C and 25°C), we found the routine metabolic rate of D. villosus to be significantly lower than that of the other studied gammarid species at the medium temperature level. The estimated resting metabolic rate indicated a similar trend. At 5°C and 25°C, both routine and resting metabolic rate did not differ between species. Compared to G. fossarum and G. roeselii, D. villosus exhibited lower locomotor activity at the low and medium temperatures (5°C and 15°C). In contrast, its locomotor activity increased at the high experimental temperature (25°C). G. fossarum and G. roeselii were apparently more active than D. villosus at all studied temperatures. We conclude that D. villosus has both physiological and behavioral adaptations that lead to a reduction in metabolic energy expenditure, which is assumed to be beneficial and might contribute to its invasive success. Copyright © 2015 Elsevier Inc. All rights reserved.

Green leaf allowance and dairy ewe performance grazing on tropical pasture.

PubMed

De Souza, J; Batistel, F; Ticiani, E; Sandri, E C; Pedreira, C G S; Oliveira, D E

2014-06-01

The objective of this study was to explain the influence of green leaf allowance levels on the performance of dairy ewes grazing a tropical grass. Seventy-two lactating ewes grazed Aruana guineagrass (Panicum maximum Jacq. cv. Aruana) for 80 d. The treatments were 4 daily levels of green leaf allowance (GLA) on a DM basis corresponding to 4, 7, 10, and 13 kg DM/100 kg BW, which were named low, medium-low, medium-high, and high level, respectively. The experimental design was completely randomized with 3 replications. During the experimental period, 4 grazing cycles were evaluated in a rotational stocking grazing method (4 d of grazing and 16 d of rest). There was a linear effect of GLA on forage mass, and increasing GLA resulted in increased total leaf mass, reaching an asymptotic plateau around the medium-high GLA level. The stem mass increased with increased GLA, and a pronounced increase was observed between medium-high and high GLAs. Increasing GLA increased both forage disappearance rate and postgrazing forage mass. Leaf proportion increased with GLA, peaking at the medium-high level, and the opposite occurred for stem proportions, which reduced until medium-high GLA level, followed by an increase on high GLA. Forage CP decreased linearly with GLA, and increasing GLA from low to high reduced CP content by 31%. On the other hand, NDF increased 14% and ADF increased 26%, both linearly in response to greater GLA levels. Total digestible nutrients decreased linearly by 8% when GLA increased from low to high level. Milk yield increased, peaking at medium-high GLA (1.75 kg ewe(-1) d(-1)) and decreased at high GLA level (1.40 kg ewe(-1) d(-1)). Milk composition was not affected by the GLA levels. There was a reduction in stocking rate from 72 to 43 ewes/ha when GLA increased from low to high level. Productivity (milk yield kg ha(-1) d(-1)) increased as GLA increased, peaking at medium-low level (115 kg ha(-1) d(-1)). Although this tropical grass showed the same pattern in responses to GLA levels as reported in the literature with temperate pastures, the magnitude of the process changed and the maximum response in milk yield from lactating dairy ewes grazing a tropical pasture would be achieved with higher forage allowances than in temperate pastures. Overall, Aruana guineagrass grazed by lactating dairy ewes should be managed to provide 7 to 10 GLA in kg DM/100 kg BW according to the production goals.

Influence of substrate exposure history on biodegradation in a porous medium

NASA Astrophysics Data System (ADS)

Park, J.; Chen, Y.-M.; Kukor, J. J.; Abriola, L. M.

2001-10-01

This study investigates the influence of fluctuating toluene concentrations on aerobic toluene degradation in a sandy porous medium colonized with Ralstonia pickettii PKO1. Column effluent toluene concentrations were found to increase after a temporary decrease in influent toluene concentration. Subsequent examination of the spatial gradient of toluene degradative activity in the column suggested that the observed increase in effluent toluene concentrations was attributable to an adverse effect of toluene limitation on the biodegradative activity of attached cells. The traditional Michaelis-Menten-type biodegradation equation associated with batch-measured Vmax (2.26 mg toluene/mg living cell/day) and KS (1.20 mg toluene/l) of nonstarved cells was unable to predict the observed toluene breakthrough behavior when the column had been previously exposed to no-toluene conditions. An alternative modeling approach was developed based upon the assumptions that (i) degradative activity was completely deactivated within the no-toluene exposure period (53.5 h) and (ii) a lag-phase was present prior to the subsequent reactivation of degradative activity in previously toluene-starved cells. These assumptions were independently verified by batch microbial investigations, and the modified model provided a good fit to the same observed toluene breakthrough curve. Application of single lag-time and threshold concentration values, however, failed to predict observed toluene breakthrough under different toluene exposure conditions. Results of this experimental and modeling investigation suggested that substrate exposure history, including the length of the starvation period and the level of substrate concentration, affected the induction of biodegradation in the porous medium.

Comparison of Impurities in Charcoal Sorbents Found by Neutron Activation Analysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Doll, Charles G.; Finn, Erin C.; Cantaloub, Michael G.

2013-01-01

Abstract: Neutron activation of gas samples in a reactor often requires a medium to retain sufficient amounts of the gas for analysis. Charcoal is commonly used to adsorb gas and hold it for activation; however, the amount of activated sodium in the charcoal after irradiation swamps most signals of interest. Neutron activation analysis (NAA) was performed on several commonly available charcoal samples in an effort to determine the activation background. The results for several elements, including the dominant sodium element, are reported. It was found that ECN charcoal had the lowest elemental background, containing sodium at 2.65 ± 0.05 ppm,more » as well as trace levels of copper and tungsten.« less

Influence of wildfires in the boreal forests of Eastern Siberia on atmospheric aerosol parameters

NASA Astrophysics Data System (ADS)

Tomshin, Oleg A.; Solovyev, Vladimir S.

2017-11-01

The results of studies of the dynamics of forest fires in the boreal forests of Yakutia (Eastern Siberia) for 2001-2016 are presented. Variations of aerosol optical thickness (AOT), aerosol index (AI) and total carbon monoxide content during May-September were studied depending on the different forest fire activity level. It is shown that the seasonal variations of AOT, AI and CO in the most fire-dangerous years differ significantly from the fire seasons when forest fire activity was medium or low.

Laser activation of a nutrient medium and antibiotic solutions and its estimation by of bacteria growth dynamics

NASA Astrophysics Data System (ADS)

Malov, Alexander N.; Neupokoeva, Anna V.; Kokorina, Lubov A.; Simonova, Elena V.

2016-11-01

A laser photomodifacation of nutrient mediums and antibiotics results at the microbiological supervision of bacteria colonies growth are discussed. It is experimentally shown, that on the irradiated media there is a delay of bacterial colonies growth number. Influence of laser radiation on activity of an antibiotic also is experimentally studied. It is revealed, that laser photomodifacation increases antimicrobic activity of a preparation. The mechanism of biological solutions activation is connected with the phenomenon laser nanoclusterization. Parameters of bacteria growth bacteria growth dynamics allow to numerically estimate degree of laser activation of nutrient mediums and pharmaceutical preparations.

Antifungal and Antiochratoxigenic Activities of Essential Oils and Total Phenolic Extracts: A Comparative Study

PubMed Central

EL Khoury, Rachelle; Atoui, Ali; Mathieu, Florence; Kawtharani, Hiba; EL Khoury, Anthony; Maroun, Richard G.; EL Khoury, Andre

2017-01-01

This study is intended to prevent ochratoxin A (OTA) production by Aspergillus carbonarius S402 using essential oils (EOs) and total phenolic compounds extracted from plants and herbs. The EOs used in this study are the following: bay leaves, cumin, fenugreek, melissa, mint, and sage. As for the phenolic compounds, they were extracted from bay leaves, cumin, fenugreek, melissa, mint, sage, anise, chamomile, fennel, rosemary, and thyme. The experiments were conducted on Synthetic Grape Medium (SGM) medium at 28 °C for 4 days. OTA was extracted from the medium with methanol and quantified using HPLC (High Performance Liquid Chromatography). Results showed that EOs had a greater impact than the total phenolic extracts on the OTA production. Reduction levels ranged between 25% (sage) and 80% (melissa) for the EOs at 5 µL mL−1, and 13% (thyme) and 69% (mint) for the phenolic extracts. Although they did not affect the growth of A. carbonarius, total phenolic extracts and EOs were capable of partially reducing OTA production. Reduction levels depended on the nature of the plants and the concentration of the EOs. Reducing OTA with natural extracts could be a solution to prevent OTA production without altering the fungal growth, thus preserving the natural microbial balance. PMID:28698493

Inhibition of AQP1 Hampers Osteosarcoma and Hepatocellular Carcinoma Progression Mediated by Bone Marrow-Derived Mesenchymal Stem Cells.

PubMed

Pelagalli, Alessandra; Nardelli, Anna; Fontanella, Raffaela; Zannetti, Antonella

2016-07-11

The complex cross-talk between tumor cells and their surrounding stromal environment plays a key role in the pathogenesis of cancer. Among several cell types that constitute the tumor stroma, bone marrow-derived mesenchymal stem cells (BM-MSCs) selectively migrate toward the tumor microenvironment and contribute to the active formation of tumor-associated stroma. Therefore, here we elucidate the involvement of BM-MSCs to promote osteosarcoma (OS) and hepatocellular carcinoma (HCC) cells migration and invasion and deepening the role of specific pathways. We analyzed the function of aquaporin 1 (AQP1), a water channel known to promote metastasis and neoangiogenes. AQP1 protein levels were analyzed in OS (U2OS) and HCC (SNU-398) cells exposed to conditioned medium from BM-MSCs. Tumor cell migration and invasion in response to BM-MSC conditioned medium were evaluated through a wound healing assay and Boyden chamber, respectively. The results showed that the AQP1 level was increased in both tumor cell lines after treatment with BM-MSC conditioned medium. Moreover, BM-MSCs-mediated tumor cell migration and invasion were hampered after treatment with AQP1 inhibitor. These data suggest that the recruitment of human BM-MSCs into the tumor microenvironment might cause OS and HCC cell migration and invasion through involvement of AQP1.

Comparison of Vibrio parahaemolyticus grown in estuarine water and rich medium.

PubMed Central

Pace, J; Chai, T J

1989-01-01

Cell envelope composition and selected physiological traits of Vibrio parahaemolyticus were studied in regard to the Kanagawa phenomenon and growth conditions. Cell envelopes were prepared from cells cultured in Proteose Peptone-beef extract (Difco Laboratories, Detroit, Mich.) medium or filtered estuarine water. Protein, phospholipid, and lipopolysaccharide contents varied with culture conditions. The phospholipids present in the cell envelopes were identified as phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin. Phosphatidylethanolamine decreased and phosphatidylglycerol increased in cells grown in estuarine water. Profiles of proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated numerous protein species, with four to six predominant proteins ranging from 26,000 to 120,000 in molecular weight. The profile of V. parahaemolyticus cell envelope proteins was unique and might be useful in the identification of the organism. Alkaline phosphatase activity was slightly higher in Kanagawa-negative strains and was higher in cells grown in estuarine water than in cells grown in rich laboratory medium. The DNA levels in estuarine water-grown cells increased, while RNA levels and cell volume decreased. Bacteriophage sensitivity typing demonstrated a close intraspecies relationship. Results indicated that Kanagawa-positive and -negative strains were closely related, but they could be grouped separately and may have undergone starvation-related physiological changes when cultured in estuarine water. Images PMID:2782869

Biologically Active Organic Matter in Soils of European Russia

NASA Astrophysics Data System (ADS)

Semenov, V. M.; Kogut, B. M.; Zinyakova, N. B.; Masyutenko, N. P.; Malyukova, L. S.; Lebedeva, T. N.; Tulina, A. S.

2018-04-01

Experimental and literature data on the contents and stocks of active organic matter in 200 soil samples from the forest-tundra, southern-taiga, deciduous-forest, forest-steppe, dry-steppe, semidesert, and subtropical zones have been generalized. Natural lands, agrocenoses, treatments of long-term field experiments (bare fallow, unfertilized and fertilized crop rotations, perennial plantations), and different layers of soil profile are presented. Sphagnum peat and humus-peat soil in the tundra and forest-tundra zones are characterized by a very high content of active organic matter (300-600 mg C/100 g). Among the zonal soils, the content of active organic matter increases from the medium (75-150 mg C/100 g) to the high (150-300 mg C/100 g) level when going from soddy-podzolic soil to gray forest and dark-gray forest soils and then to leached chernozem. In the series from typical chernozem to ordinary and southern chernozem and chestnut and brown semidesert soils, a decrease in the content of active organic matter to the low (35-75 mg C/100 g) and very low (<35 mg C/100 g) levels is observed. Acid brown forest soil in the subtropical zone is characterized by a medium supply with active organic matter. Most arable soils are mainly characterized by low or very low contents of active organic matter. In the upper layers of soils, active organic matter makes up 1.2-11.1% of total Corg. The profile distribution of active organic matter in the studied soils coincides with that of Corg: their contents appreciably decrease with depth, except for brown semidesert soil. The stocks of active organic matter vary from 0.4 to 5.4 t/ha in the layer of 0-20 cm and from 1.0 to 12.4/ha in the layer of 0-50 cm of different soil types.

[Role of calcium ions in the mechanism of action of acetylcholine on energy metabolism in rat liver mitochondria].

PubMed

Vatamaniuk, M Z; Artym, V V; Kuka, O B; Doliba, M M; Shostakovs'ka, I V

1996-01-01

It is shown that administration of acetylcholine to animals (50 micrograms per 100 g of body weight) leads to the activation of respiration and oxidative phosphorylation in the rat liver mitochondria under oxidation of alpha-ketoglutarate; this effect depends on the concentration of calcium ions in the incubation medium of mitochondria. The rate of ADP-stimulated respiration of mitochondria of experimental animals reaches its maximum level under lower concentrations of Ca2+ than in the control animals. The results of investigation of dependence of acetyl choline effect on respiration of mitochondria on the concentration of alpha-ketoglutarate in calcium and calcium-free incubation medium have shown that the half-maximum effect of acetylcholine is observed in calcium medium at lower concentration of the substrate than in calcium-free medium. The latter indicates to the increase of affinity of alpha-ketoglutarate dehydrogenase to alpha-ketoglutarate under these conditions. It is found out that acetylcholine (1.10(-8) M) increases the rate of ADP- and Ca(2+)-stimulated respiration of mitochondria of isolated perfused rat liver, while mutual effect of verapamyl and niphedipin removes this effect.

Antidiabetic activity of medium-polar extract from the leaves of Stevia rebaudiana Bert. (Bertoni) on alloxan-induced diabetic rats

PubMed Central

Misra, Himanshu; Soni, Manish; Silawat, Narendra; Mehta, Darshana; Mehta, B. K.; Jain, D. C.

2011-01-01

Objective: To investigate the medicative effects of medium-polar (benzene:acetone, 1:1, v/v) extract of leaves from Stevia rebaudiana (family Asteraceae) on alloxan-induced diabetic rats. Materials and Methods: Diabetes was induced in adult albino Wistar rats by intraperitoneal (i.p.) injection of alloxan (180 mg/kg). Medium-polar extract was administered orally at daily dose of 200 and 400 mg/kg body wt. basis for 10 days. The control group received normal saline (0.9%) for the same duration. Glibenclamide was used as positive control reference drug against Stevia extract. Results: Medium-polar leaf extract of S. rebaudiana (200 and 400 mg/kg) produced a delayed but significant (P < 0.01) decrease in the blood glucose level, without producing condition of hypoglycemia after treatment, together with lesser loss in the body weight as compared with standard positive control drug glibenclamide. Conclusions: Treatment of diabetes with sulfonylurea drugs (glibenclamide) causes hypoglycemia followed by greater reduction in body weight, which are the most worrisome effects of these drugs. Stevia extract was found to antagonize the necrotic action of alloxan and thus had a re-vitalizing effect on β-cells of pancreas. PMID:21687353

Pseudomonas aeruginosa mutants resistant to urea inhibition of growth on acetanilide.

PubMed

Gregoriou, M; Brown, P R; Tata, R

1977-11-01

Pseudomonas aeruginosa AI 3 was able to grow in medium containing acetanilide (N-phenylacetamide) as a carbon source when NH4+ was the nitrogen source but not when urea was the nitrogen source. AIU mutants isolated from strain AI 3 grew on either medium. Urease levels in bacteria grown in the presence of urea were 10-fold lower when NH4+ or acetanilide was also in the medium, but there were no apparent differences in urease or its synthesis between strain AI 3 and mutant AIU 1N. The first metabolic step in the acetanilide utlization is catalyzed by an amidase. Amidases in several AIU strains showed altered physiochemical properties. Urea inhibited amidase in a time-dependent reaction, but the rates of the inhibitory reaction with amidases from the AIU mutants were slower than with AI 3 amidase. The purified amidase from AIU 1N showed a marked difference in its pH/activity profile from that obtained with purified AI 3 amidase. These observations indicate that the ability of strain AIU 1N and the other mutants to grow on acetanilide/urea medium is associated with a mutation in the amidase structural gene; this was confirmed for strain AIU 1N by transduction.

Effects of sulfate concentrations on the expression of a soybean seed storage protein gene and its reversibility in transgenic Arabidopsis thaliana.

PubMed

Hirai, M Y; Fujiwara, T; Chino, M; Naito, S

1995-10-01

Transgenic expression of genes encoding the alpha' and beta subunits of beta-conglycinin, one of the major seed storage proteins of soybean (Glycine max [L.] Merr.), was analyzed in Arabidopsis thaliana (L.) Heynh. under conditions of sulfate deficiency. Temporal patterns of expression of both the intact beta subunit gene and the beta subunit gene promoter fused to the beta-glucuronidase (GUS) gene are similar in soil-less cultures using rockwool, suggesting that the response to sulfate deficiency is regulated mainly at the level of transcription. In hydroponic cultures with various concentrations of sulfate, expression of both the intact beta subunit gene and the beta subunit gene promoter-GUS fusion gene were negatively correlated to increased sulfate concentrations in the culture medium. Transfer of transgenic A. thaliana plants carrying the beta subunit gene promoter-GUS fusion from sulfate-deficient to sulfate-sufficient control medium caused GUS activity in developing siliques to be repressed within two days. A reverse shift, where the plants were transferred from the control to sulfate-deficient medium, caused GUS activity to become higher than that in seeds of the control plants within two days. These results indicate that the expression of the beta subunit gene promoter responds rapidly to changes of sulfate availability.

Metabolic response to MMS-mediated DNA damage in Saccharomyces cerevisiae is dependent on the glucose concentration in the medium.

PubMed

Kitanovic, Ana; Walther, Thomas; Loret, Marie Odile; Holzwarth, Jinda; Kitanovic, Igor; Bonowski, Felix; Van Bui, Ngoc; Francois, Jean Marie; Wölfl, Stefan

2009-06-01

Maintenance and adaptation of energy metabolism could play an important role in the cellular ability to respond to DNA damage. A large number of studies suggest that the sensitivity of cells to oxidants and oxidative stress depends on the activity of cellular metabolism and is dependent on the glucose concentration. In fact, yeast cells that utilize fermentative carbon sources and hence rely mainly on glycolysis for energy appear to be more sensitive to oxidative stress. Here we show that treatment of the yeast Saccharomyces cerevisiae growing on a glucose-rich medium with the DNA alkylating agent methyl methanesulphonate (MMS) triggers a rapid inhibition of respiration and enhances reactive oxygen species (ROS) production, which is accompanied by a strong suppression of glycolysis. Further, diminished activity of pyruvate kinase and glyceraldehyde-3-phosphate dehydrogenase upon MMS treatment leads to a diversion of glucose carbon to glycerol, trehalose and glycogen accumulation and an increased flux through the pentose-phosphate pathway. Such conditions finally result in a significant decline in the ATP level and energy charge. These effects are dependent on the glucose concentration in the medium. Our results clearly demonstrate that calorie restriction reduces MMS toxicity through increased respiration and reduced ROS accumulation, enhancing the survival and recovery of cells.

Effect of body mass and clothing on decomposition of pig carcasses.

PubMed

Matuszewski, Szymon; Konwerski, Szymon; Frątczak, Katarzyna; Szafałowicz, Michał

2014-11-01

Carcass mass and carcass clothing are factors of potential high forensic importance. In casework, corpses differ in mass and kind or extent of clothing; hence, a question arises whether methods for post-mortem interval estimation should take these differences into account. Unfortunately, effects of carcass mass and clothing on specific processes in decomposition and related entomological phenomena are unclear. In this article, simultaneous effects of these factors are analysed. The experiment followed a complete factorial block design with four levels of carcass mass (small carcasses 5-15 kg, medium carcasses 15.1-30 kg, medium/large carcasses 35-50 kg, large carcasses 55-70 kg) and two levels of carcass clothing (clothed and unclothed). Pig carcasses (N = 24) were grouped into three blocks, which were separated in time. Generally, carcass mass revealed significant and frequently large effects in almost all analyses, whereas carcass clothing had only minor influence on some phenomena related to the advanced decay. Carcass mass differently affected particular gross processes in decomposition. Putrefaction was more efficient in larger carcasses, which manifested itself through earlier onset and longer duration of bloating. On the other hand, active decay was less efficient in these carcasses, with relatively low average rate, resulting in slower mass loss and later onset of advanced decay. The average rate of active decay showed a significant, logarithmic increase with an increase in carcass mass, but only in these carcasses on which active decay was driven solely by larval blowflies. If a blowfly-driven active decay was followed by active decay driven by larval Necrodes littoralis (Coleoptera: Silphidae), which was regularly found in medium/large and large carcasses, the average rate showed only a slight and insignificant increase with an increase in carcass mass. These results indicate that lower efficiency of active decay in larger carcasses is a consequence of a multi-guild and competition-related pattern of this process. Pattern of mass loss in large and medium/large carcasses was not sigmoidal, but rather exponential. The overall rate of decomposition was strongly, but not linearly, related to carcass mass. In a range of low mass decomposition rate increased with an increase in mass, then at about 30 kg, there was a distinct decrease in rate, and again at about 50 kg, the rate slightly increased. Until about 100 accumulated degree-days larger carcasses gained higher total body scores than smaller carcasses. Afterwards, the pattern was reversed; moreover, differences between classes of carcasses enlarged with the progress of decomposition. In conclusion, current results demonstrate that cadaver mass is a factor of key importance for decomposition, and as such, it should be taken into account by decomposition-related methods for post-mortem interval estimation.

Stimulation of plasmin activity in cultured human fibroblast cells by Porphyromonas endodontalis.

PubMed

Oikawa, T; Ogura, N; Akiba, M; Abiko, Y; Takiguchi, H; Izumi, H

1993-09-01

1. Plasmin activity in the conditioned medium of Gin-1 cells, a human gingival fibroblast cell line, was stimulated by Porphyromonas endodontalis, a putative pathogen of oral submucous abscesses, in a time- and dose-dependent manner. 2. P. endodontalis stimulated the activity of plasminogen activator in both the conditioned medium and the cell lysate. The plasminogen activator in Gin-1 cells was approx. 50 kDa by zymography. 3. The conditioned medium of Gin-1 cells exposed to P. endodontalis stimulated the conversion of human serum prekallikrein to kallikrein. 4. These results suggested that P. endodontalis stimulates the plasminogen activator-plasmin system in Gin-1 cells, and that activated plasmin plays a role in the progress of periodontal tissue inflammation.

The "Ideal" vs. the "Reality": Medium of Instruction Policy and Implementation in Different Class Levels in a Western Kenyan School

ERIC Educational Resources Information Center

Jones, Jennifer M.

2014-01-01

This paper investigates the perceptions of teachers from one Sabaot dominated primary school in western Kenya regarding the medium of instruction (MoI) policy in different class levels. While their "ideal" MoI policy bears some resemblance to the official policy which advocates mother tongue (MT) as medium in lower primary and English in…

Identification and characterisation of xylanolytic yeasts isolated from decaying wood and sugarcane bagasse in Brazil.

PubMed

Lara, Carla A; Santos, Renata O; Cadete, Raquel M; Ferreira, Carla; Marques, Susana; Gírio, Francisco; Oliveira, Evelyn S; Rosa, Carlos A; Fonseca, César

2014-06-01

In this study, yeasts associated with lignocellulosic materials in Brazil, including decaying wood and sugarcane bagasse, were isolated, and their ability to produce xylanolytic enzymes was investigated. A total of 358 yeast isolates were obtained, with 198 strains isolated from decaying wood and 160 strains isolated from decaying sugarcane bagasse samples. Seventy-five isolates possessed xylanase activity in solid medium and were identified as belonging to nine species: Candida intermedia, C. tropicalis, Meyerozyma guilliermondii, Scheffersomyces shehatae, Sugiyamaella smithiae, Cryptococcus diffluens, Cr. heveanensis, Cr. laurentii and Trichosporon mycotoxinivorans. Twenty-one isolates were further screened for total xylanase activity in liquid medium with xylan, and five xylanolytic yeasts were selected for further characterization, which included quantitative analysis of growth in xylan and xylose and xylanase and β-D-xylosidase activities. The yeasts showing the highest growth rate and cell density in xylan, Cr. laurentii UFMG-HB-48, Su. smithiae UFMG-HM-80.1 and Sc. shehatae UFMG-HM-9.1a, were, simultaneously, those exhibiting higher xylanase activity. Xylan induced the highest level of (extracellular) xylanase activity in Cr. laurentii UFMG-HB-48 and the highest level of (intracellular, extracellular and membrane-associated) β-D-xylosidase activity in Su. smithiae UFMG-HM-80.1. Also, significant β-D-xylosidase levels were detected in xylan-induced cultures of Cr. laurentii UFMG-HB-48 and Sc. shehatae UFMG-HM-9.1a, mainly in extracellular and intracellular spaces, respectively. Under xylose induction, Cr. laurentii UFMG-HB-48 showed the highest intracellular β-D-xylosidase activity among all the yeast tested. C. tropicalis UFMG-HB 93a showed its higher (intracellular) β-D-xylosidase activity under xylose induction and higher at 30 °C than at 50 °C. This study revealed different xylanolytic abilities and strategies in yeasts to metabolise xylan and/or its hydrolysis products (xylo-oligosaccharides and xylose). Xylanolytic yeasts are able to secrete xylanolytic enzymes mainly when induced by xylan and present different strategies (intra- and/or extracellular hydrolysis) for the metabolism of xylo-oligosaccharides. Some of the unique xylanolytic traits identified here should be further explored for their applicability in specific biotechnological processes.

Divergent calcium signaling in RBCs from Tropidurus torquatus (Squamata – Tropiduridae) strengthen classification in lizard evolution

PubMed Central

Beraldo, Flávio H; Garcia, Célia RS

2007-01-01

Background We have previously reported that a Teiid lizard red blood cells (RBCs) such as Ameiva ameiva and Tupinambis merianae controls intracellular calcium levels by displaying multiple mechanisms. In these cells, calcium stores could be discharged not only by: thapsigargin, but also by the Na+/H+ ionophore monensin, K+/H+ ionophore nigericin and the H+ pump inhibitor bafilomycin as well as ionomycin. Moreover, these lizards possess a P2Y-type purinoceptors that mobilize Ca2+ from intracellular stores upon ATP addition. Results Here we report, that RBCs from the tropidurid lizard Tropidurus torquatus store Ca2+ in endoplasmic reticulum (ER) pool but unlike in the referred Teiidae, these cells do not store calcium in monensin-nigericin sensitive pools. Moreover, mitochondria from T. torquatus RBCs accumulate Ca2+. Addition of ATP to a calcium-free medium does not increase the [Ca2+]c levels, however in a calcium medium we observe an increase in cytosolic calcium. This is an indication that purinergic receptors in these cells are P2X-like. Conclusion T. torquatus RBCs present different mechanisms from Teiid lizard red blood cells (RBCs), for controlling its intracellular calcium levels. At T. torquatus the ion is only stored at endoplasmic reticulum and mitochondria. Moreover activation of purinergic receptor, P2X type, was able to induce an influx of calcium from extracelullar medium. These studies contribute to the understanding of the evolution of calcium homeostasis and signaling in nucleated RBCs. PMID:17716375

Divergent calcium signaling in RBCs from Tropidurus torquatus (Squamata--Tropiduridae) strengthen classification in lizard evolution.

PubMed

Beraldo, Flávio H; Garcia, Célia R S

2007-08-23

We have previously reported that a Teiid lizard red blood cells (RBCs) such as Ameiva ameiva and Tupinambis merianae controls intracellular calcium levels by displaying multiple mechanisms. In these cells, calcium stores could be discharged not only by: thapsigargin, but also by the Na+/H+ ionophore monensin, K+/H+ ionophore nigericin and the H+ pump inhibitor bafilomycin as well as ionomycin. Moreover, these lizards possess a P2Y-type purinoceptors that mobilize Ca2+ from intracellular stores upon ATP addition. Here we report, that RBCs from the tropidurid lizard Tropidurus torquatus store Ca2+ in endoplasmic reticulum (ER) pool but unlike in the referred Teiidae, these cells do not store calcium in monensin-nigericin sensitive pools. Moreover, mitochondria from T. torquatus RBCs accumulate Ca2+. Addition of ATP to a calcium-free medium does not increase the [Ca2+]c levels, however in a calcium medium we observe an increase in cytosolic calcium. This is an indication that purinergic receptors in these cells are P2X-like. T. torquatus RBCs present different mechanisms from Teiid lizard red blood cells (RBCs), for controlling its intracellular calcium levels. At T. torquatus the ion is only stored at endoplasmic reticulum and mitochondria. Moreover activation of purinergic receptor, P2X type, was able to induce an influx of calcium from extracellular medium. These studies contribute to the understanding of the evolution of calcium homeostasis and signaling in nucleated RBCs.

The influence of physical activity on hair toxic and essential trace element content in male and female students.

PubMed

Zaitseva, Irina P; Skalny, Andrey A; Tinkov, Alexey A; Berezkina, Elena S; Grabeklis, Andrei R; Skalny, Anatoly V

2015-02-01

The primary aim of the current study is to estimate the effect of different physical activity levels on hair trace element content in male and female students. A total of 113 students (59 women and 54 men) of P. G. Demidov Yaroslavl State University (Yaroslavl, Russia) took part in the current investigation. According to the level of the physical activity, all students were divided into three groups: high, medium, and low physical activity. Essential and toxic metal content (μg/g) in hair samples was assessed by inductively coupled plasma mass spectrometry using NexION 300D + NWR213 (Perkin-Elmer, USA). The obtained data show that hair iodine, zinc, arsenic, nickel, and tin levels are not related to physical activity in male and female students. At the same time, increased physical activity is associated with decreased hair copper, vanadium, bismuth, and mercury content in comparison to the low physical activity groups. Students with higher physical activity are also characterized by significantly higher hair cobalt, iron, manganese, selenium, cadmium, lithium, and lead concentrations. Finally, statistical analysis has revealed maximal gender differences in hair trace element content in the high physical activity groups, whereas in the low activity groups, the hair metal concentrations were nearly similar in females and males.

Consumer preferences for pig welfare - Can the market accommodate more than one level of welfare pork?

PubMed

Denver, Sigrid; Sandøe, Peter; Christensen, Tove

2017-07-01

The purpose of the present paper is to investigate the market potential of pork labelled to indicate medium and high levels of animal welfare. The paper asks, in particular, whether there is a risk that Danish consumers will abandon high level welfare pork if less expensive products with a medium level of animal welfare became available. The study was based on an online questionnaire with a choice experiment involving 396 Danish respondents. The results indicated that the Danish market could accommodate more than one pork product with a welfare label but the price differential separating medium and high level animal welfare pork will have to be quite narrow. In addition, full willingness-to-pay of consumers who want to buy high level welfare pork cannot be relied upon to incentivise new consumers to buy medium welfare pork. Further, raising brand awareness in the shopping situation and improving consumer's understanding of brand attributes for high level welfare brands were found to be vital. Copyright © 2017 Elsevier Ltd. All rights reserved.

Predicting levels of Latino depression: acculturation, acculturative stress, and coping.

PubMed

Torres, Lucas

2010-04-01

Past research has noted that aspects of living in the United States place Latinos at risk for experiencing psychological problems. However, the specific features of the adaptation process that contribute to depression remain unclear. The purpose of the present study was to investigate the ability of acculturation, acculturative stress, and coping to predict membership into low, medium, and high groups of depression among Latinos. Within a group of 148 Latino adults from the community, a multinomial logistic regression revealed that an Anglo orientation, English competency pressures, and active coping differentiated high from low depression and that a Latino orientation and, to some extent, the pressure to acculturate distinguished medium from low depression. These results highlight a pattern of characteristics that function as risk and protective factors in relation to level of symptom severity. The findings are discussed in terms of implications for Latino mental health, including considerations for intervention and prevention. PsycINFO Database Record (c) 2010 APA, all rights reserved.

The Relationship between Performance on Mathematical Word Problems and Language Proficiency for Students Learning through the Medium of Irish

ERIC Educational Resources Information Center

Riordain, Maire Ni; O'Donoghue, John

2009-01-01

Ireland has two official languages--Gaeilge (Irish) and English. Similarly, primary- and second-level education can be mediated through the medium of Gaeilge or through the medium of English. This research is primarily focused on students (Gaeilgeoiri) in the transition from Gaeilge-medium mathematics education to English-medium mathematics…

Ability to return to sports 5 years after matrix-associated autologous chondrocyte transplantation in an average population of active patients.

PubMed

Zak, Lukas; Aldrian, Silke; Wondrasch, Barbara; Albrecht, Christian; Marlovits, Stefan

2012-12-01

Cartilage injuries often occur during sports activities, and return to sports after cartilage surgery is an important outcome parameter for different treatment methods in the competitive as well as the recreationally active population. At the time of midterm follow-up after matrix-associated autologous chondrocyte transplantation (MACT), return to recreational sports at the preinjury level will be possible. Case series; Level of evidence, 4. Seventy patients (51 men, 19 women; age [mean ± standard deviation], 34.9 ± 8.6 y; range, 18-55 y) were clinically evaluated 5 years after MACT through subjective clinical scores such as the Knee Injury and Osteoarthritis Outcome Score (KOOS) sport and recreation subscales, the Tegner activity scale, and the Noyes sports activity rating scale. The level of sports participation was included in the investigation. The results 5 years after MACT showed mean values of 60.1 for the KOOS-sport, 67.4 for the Noyes, and 3.8 for the Tegner scores, meaning that regular sports activity such as cycling or running on flat ground, as well as medium-level manual labor, is possible. We noted that 74.3% of our patients returned to at least their preinjury sports level. Midterm postoperative results after MACT show that in a moderately active population, participation in regular sports is possible for most patients, at least at their preinjury recreational level and intensity, and there is a good rate of return to sports.

Physical activity energy expenditure in Dutch adolescents: contribution of active transport to school, physical education, and leisure time activities.

PubMed

Slingerland, Menno; Borghouts, Lars B; Hesselink, Matthijs K C

2012-05-01

Detailed knowledge about physical activity energy expenditure (PAEE) can guide the development of school interventions aimed at reducing overweight in adolescents. However, relevant components of PAEE have never been objectively quantified in this population. This study investigated the contribution of active transport to and from school, physical education (PE), and leisure time activities to total PAEE during a regular school week in adolescents. Seventy-three adolescents (mean age: 15.7 years) wore an individually calibrated combined heart rate-acceleration monitor and kept an activity diary during a regular school week. Branched equation modeling was used to calculate PAEE of the specific activity categories, and their relative contribution to total PAEE was determined. Active transport and PE contributed 30.0% and 17.4%, respectively, to school-related PAEE. Active transport to and from school contributed 15% to total PAEE. Youth with a high physical activity level (PAL) spent 4 hours less in sedentary behavior than subjects with a medium or low PAL (F = 77.415 (2.70), p < .001) and had higher PAEE during leisure time sports (F = 9.135 (2.70), p < .001) and work-related activities (F = 10.583 (2.70), p < .001) than youth with medium or low PAL values. Active transport and PE contribute significantly to PAEE during school hours in adolescents. To achieve an increase in total PAEE in the least active group of adolescents, promising strategies might be to reduce inactive behavior, increase participation in leisure time sports, and possibly to replace inactive for active jobs. © 2012, American School Health Association.

Osmotic control of glycine betaine biosynthesis and degradation in Rhizobium meliloti

DOE Office of Scientific and Technical Information (OSTI.GOV)

Smith, L.T.; Pocard, J.A.; Bernard, T.

1988-07-01

Intracellular accumulation of glycine betaine has been shown to confer an enhanced level of osmotic stress tolerance in Rhizobium meliloti. In this study, the authors used a physiological approach to investigate the mechanism by which glycine betaine is accumulated in osmotically stressed R. meliloti. Results from growth experiments, /sup 14/C labeling of intermediates, and enzyme activity assays are presented. The results provide evidence for the pathway of biosynthesis and degradation of glycine betaine and the osmotic effects on this pathway. High osmolarity in the medium decreased the activities of the enzymes involved in the degradation of glycine betaine but notmore » those of enzymes that lead to its biosynthesis from choline. Thus, the concentration of the osmoprotectant glycine betaine is increased in stressed cells. This report demonstrates the ability of the osmolarity of the growth medium to regulate the use of glycine betaine as a carbon and nitrogen source or as an osmoprotectant. The mechanisms of osmoregulation in R. meliloti and Escherichia coli are compared.« less

Production, thermal stability and immobilisation of inulinase from Fusarium oxysporum.

PubMed

Gupta, A K; Rathore, P; Kaur, N; Singh, R

1990-01-01

Fusarium oxysporum produced maximum extracellular inulinase after 9 days of its growth at 25 degrees C on a medium (pH 5.5) containing 3% fructan and 0.2% sodium nitrate. The level of this enzyme decreased on the addition of either glucose, fructose, galactose or sucrose to F. oxysporum already growing on a fructan-containing medium. A significant increase in invertase production which resulted in an increase of the invertase/inulinase (S/I) ratio, was observed on addition of inulin to this fungus growing on other carbon sources. Glycerol (10%) gave better protection to inulinase against thermal denaturation at 50 degrees C compared to ethylene glycol and sorbitol. Inulinase immobilised in polyacrylamide gel retained 45% of its original activity. The immobilised enzyme showed a higher optimum temperature (45 degrees C) compared to free enzyme (37 degrees C). The immobilised enzyme after storage at 25 degrees C for 96 h showed 58% activity. Thermal stability of entrapped inulinase increased in the presence of inulin.

Variation in Hydraulic Conductivity with Decreasing pH in a Biologically-Clogged Porous Medium

NASA Astrophysics Data System (ADS)

Kirk, M. F.; Santillan, E.; McGrath, L. K.; Altman, S. J.

2011-12-01

Biological clogging can significantly lower the hydraulic conductivity of porous media, potentially helping to limit CO2 transport from geological carbon storage reservoirs. How clogging is affected by CO2 injection, however, is unclear. We used column experiments to examine how decreasing pH, a geochemical change associated with CO2 injection, will affect the hydraulic conductivity (K) of biologically clogged porous medium. Four biologically-active experiments and two control experiments were performed. Columns consisted of 1 mm2 capillary tubes filled with 105-150 μm diameter glass beads. Artificial groundwater medium containing 1 mM glucose was pumped through the columns at a rate of 0.015 mL/min (q = 21.6 m/day; Re = 0.045). Each column was inoculated with 10^8 CFU of Pseudomonas fluorescens tagged with a green fluorescent protein; cells introduced to control columns were heat sterilized. Biomass distribution and transport was monitored using scanning laser confocal microscopy and effluent plating. Growth was allowed to occur for 5 days in medium with pH 7 in the biologically active columns. During that time, K decreased to values ranging from 10 to 27% of the average control K and effluent cell levels increased to about 10^8 CFU/mL. Next, the pH of the inflowing medium was lowered to 4 in three experiments and 5.5 in one experiment. After pH 4 medium was introduced, K increased to values ranging from 21 to 64% of the average control K and culturable cell levels in the effluent fell by about 4 log units. Confocal images show that clogging persisted in the columns at pH 4 because most of the microbial biomass remained attached to bead surfaces. In the experiment where pH was lowered to 5.5, K changed little because biological clogging remained entirely intact. The concentration of culturable cells in the effluent was also invariant. These results suggest that biomass in porous medium will largely remain in place following exposure to acidic water in a CO2 storage reservoir, particularly where buffering is able to limit the extent of acidification. This material is based upon work supported as part of the Center for Frontiers of Subsurface Energy Security, an Energy Frontier Research Center funded by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under Award Number DE-SC0001114. Sandia National Laboratories is a multi-program laboratory managed and operated by Sandia Corporation, a wholly owned subsidiary of Lockheed Martin Corporation, for the U.S. Department of Energy's National Nuclear Security Administration under contract DE-AC04-94AL85000.

Naringin Stimulates Osteogenic Differentiation of Rat Bone Marrow Stromal Cells via Activation of the Notch Signaling Pathway

PubMed Central

Yu, Guo-yong; Zheng, Gui-zhou; Chang, Bo; Hu, Qin-xiao; Lin, Fei-xiang; Liu, De-zhong; Wu, Chu-cheng; Du, Shi-xin

2016-01-01

Naringin is a major flavonoid found in grapefruit and is an active compound extracted from the Chinese herbal medicine Rhizoma Drynariae. Naringin is a potent stimulator of osteogenic differentiation and has potential application in preventing bone loss. However, the signaling pathway underlying its osteogenic effect remains unclear. We hypothesized that the osteogenic activity of naringin involves the Notch signaling pathway. Rat bone marrow stromal cells (BMSCs) were cultured in osteogenic medium containing-naringin, with or without DAPT (an inhibitor of Notch signaling), the effects on ALP activity, calcium deposits, osteogenic genes (ALP, BSP, and cbfa1), adipogenic maker gene PPARγ2 levels, and Notch expression were examined. We found that naringin dose-dependently increased ALP activity and Alizarin red S staining, and treatment at the optimal concentration (50 μg/mL) increased mRNA levels of osteogenic genes and Notch1 expression, while decreasing PPARγ2 mRNA levels. Furthermore, treatment with DAPT partly reversed effects of naringin on BMSCs, as judged by decreases in naringin-induced ALP activity, calcium deposits, and osteogenic genes expression, as well as upregulation of PPARγ2 mRNA levels. These results suggest that the osteogenic effect of naringin partly involves the Notch signaling pathway. PMID:27069482

2',5'-Dihydroxychalcone down-regulates endothelial connexin43 gap junctions and affects MAP kinase activation.

PubMed

Lee, Yi-Nan; Yeh, Hung-I; Tian, Tin-Yi; Lu, Wen-Wei; Ko, Yu-Shien; Tsai, Cheng-Ho

2002-09-30

We examined the effect of 2',5'-dihydroxychalcone on connexin43 (Cx43) expression and gap-junctional communication in human umbilical vein endothelial cells (HUVEC). The result showed that expression of Cx43 is rapidly reduced by 2',5'-dihydroxychalcone in a dose-dependent manner, Concomitantly, the communication function, determined by fluorescence recovery after photobleaching (FRAP), is decreased. We further investigated whether the mitogen-activated protein (MAP) kinase and the degradation pathway of gap junctions are involved in these processes. Although the change of Cx43 is not affected by the level of fetal calf serum (FCS) used in the medium, activation of MAP kinase varies, depending on the FCS level. At a low level (0.5%), the chalcone inhibits the activation, like PD98059, a specific inhibitor of MAP kinase kinase. However, at a high level (20%), MAP kinase is activated. On the other hand, the chalcone's down-regulating effect on Cx43, while is totally blocked by protease inhibitors leupeptin and N-acetyl-leucyl-norleucinal (ALLN), persists in the presence of PD98059, We concluded that 2',5'-dihydroxychalcone down-regulates Cx43 expression and gap-junctional communication in the HUVEC via enhancement of the proteolysis pathway, and this compound possesses dual effects on MAP kinase activation.

Nitrate-nitrogen levels in rural drinking water: Is there an association with age-related macular degeneration?

PubMed

Klein, Barbara E K; McElroy, Jane A; Klein, Ronald; Howard, Kerri P; Lee, Kristine E

2013-01-01

We examined the association of nitrate-nitrogen exposure from rural private drinking water and incidence of age-related macular degeneration (AMD). All participants in the Beaver Dam Eye Study (53916 improvement plan code) completed a questionnaire and had an ocular examination including standardized, graded fundus photographs at five examinations. Only information from rural residents in that study are included in this report. Data from an environmental monitoring study with probabilistic-based agro-chemical sampling, including nitrate-nitrogen, of rural private drinking water were available. Incidence of early AMD was associated with elevated nitrate-nitrogen levels in rural private drinking water supply (10.0% for low, 19.2% for medium, and 26.1% for high nitrate-nitrogen level in the right eye). The odds ratios (ORs) were 1.77 (95% confidence interval [CI]: 1.12-2.78) for medium and 2.88 (95% CI: 1.59-5.23) for high nitrate-nitrogen level. Incidence of late AMD was increased for those with medium or high levels of nitrate-nitrogen compared to low levels (2.3% for low and 5.1% for the medium or high nitrate-nitrogen level, for the right eye). The OR for medium or high nitrate-nitrogen groups was 2.80 (95% CI: 1.07-7.31) compared to the low nitrate-nitrogen group.

Nitrate-Nitrogen Levels in Rural Drinking Water: Is There an Association with Age-related Macular Degeneration?

PubMed Central

Klein, Barbara E. K.; McElroy, Jane A.; Klein, Ronald; Howard, Kerri P.; Lee, Kristine E.

2014-01-01

We examined the association of nitrate-nitrogen exposure from rural private drinking water and incidence of age-related macular degeneration (AMD). Participants of the Beaver Dam Eye Study living in rural areas within the 53916 zone improvement plan code but outside the city limits of Beaver Dam, Wisconsin (Beaver Dam Township) completed a questionnaire and ocular examination including standardized, graded fundus photographs at five examinations. Data from an environmental monitoring study with probabilistic-based agro-chemical sampling, including nitrate-nitrogen, of rural private drinking water were available. Incidence of early AMD was associated with elevated nitrate-nitrogen levels in rural private drinking water supply (10.0% for low, 19.2% for medium, and 26.1% for high nitrate-nitrogen level in the right eye). The odds ratios (ORs) were 1.77 (95% confidence interval [CI]: 1.12–2.78) for medium and 2.88 (95% CI: 1.59–5.23) for high nitrate-nitrogen level. Incidence of late AMD was increased for those with medium or high levels of nitrate-nitrogen compared to low levels (2.3% for low and 5.1% for the medium or high nitrate-nitrogen level, for the right eye). The OR for medium or high nitrate-nitrogen groups was 2.80 (95% CI: 1.07–7.31) compared to the low nitrate-nitrogen group. PMID:24007430

Work Programme, 2003.

ERIC Educational Resources Information Center

European Centre for the Development of Vocational Training, Thessaloniki (Greece).

This publication presents work program 2003 in light of the new medium-term priorities 2003-06 and activities 2003 as a consistent and systematic translation of this multiannual approach, specifying activities and outputs to meet the medium-term strategy. A main strategic activity is described--further development of a knowledge management system…

Pleiotropic role of the Sco1/SenC family copper chaperone in the physiology of Streptomyces

PubMed Central

Fujimoto, Masahiro; Yamada, Akio; Kurosawa, Junpei; Kawata, Akihiro; Beppu, Teruhiko; Takano, Hideaki; Ueda, Kenji

2012-01-01

Summary Antibiotic production and cell differentiation in Streptomyces is stimulated by micromolar levels of Cu2+. Here, we knocked out the Sco1/SenC family copper chaperone (ScoC) encoded in the conserved gene cluster ‘sco’ (the S treptomycescopper utilization) in Streptomyces coelicolor A3(2) and S. griseus. It is known that the Sco1/SenC family incorporates Cu2+ into the active centre of cytochrome oxidase (cox). The knockout caused a marked delay in antibiotic production and aerial mycelium formation on solid medium, temporal pH decline in glucose‐containing liquid medium, and significant reduction of cox activity in S. coelicolor. The scoC mutant produced two‐ to threefold higher cellular mass of the wild type exhibiting a marked cox activity in liquid medium supplied with 10 µM CuSO4, suggesting that ScoC is involved in not only the construction but also the deactivation of cox. The scoC mutant was defective in the monoamine oxidase activity responsible for cell aggregation and sedimentation. These features were similarly observed with regard to the scoC mutant of S. griseus. The scoC mutant of S. griseus was also defective in the extracellular activity oxidizing N,N′‐dimethyl‐p‐phenylenediamine sulfate. Addition of 10 µM CuSO4 repressed the activity of the conserved promoter preceding scoA and caused phenylalanine auxotrophy in some Streptomyces spp. probably because of the repression of pheA; pheA encodes prephenate dehydratase, which is located at the 3′ terminus of the putative operon structure. Overall, the evidence indicates that Sco is crucial for the utilization of copper under a low‐copper condition and for the activation of the multiple Cu2+‐containing oxidases that play divergent roles in the complex physiology of Streptomyces. PMID:22117562

Optimization of culture conditions and medium composition for the production of micrococcin GO5 by Micrococcus sp. GO5.

PubMed

Kim, Mi-Hee; Kong, Yoon-Jung; Baek, Hong; Hyun, Hyung-Hwan

2006-01-02

To enhance the production of micrococcin GO5, a bacteriocin produced by Micrococcus sp. GO5, cultivation conditions and medium composition were optimized. The optimal initial pH and temperature for bacteriocin production were 7.0-9.0 and 37 degrees C, respectively. Micrococcus sp. GO5 displayed the highest micrococcin GO5 activity when grown in modified MRS medium that contained lactose or sucrose, rather than glucose, as a carbon source. The maximum bacteriocin activity was obtained in modified MRS medium containing 0.5% tryptone and 1.0% yeast extract as nitrogen sources instead of the other nitrogen sources present in MRS medium. Bacteriocin production was greatly affected by the concentration of K(2)HPO(4); strain GO5 produced eight-fold more bacteriocin in medium containing 2.0-2.5% K(2)HPO(4) than in medium containing 0.2% K(2)HPO(4). The optimal concentration of MgSO(4).7H(2)O for bacteriocin production was 0.5%. The production of micrococcin GO5 was increased 32-fold in shake flask culture and 16-fold in a bioreactor using the optimized medium (TY medium), compared with culturing in MRS medium.

Using semantic technologies and the OSU ontology for modelling context and activities in multi-sensory surveillance systems

NASA Astrophysics Data System (ADS)

Gómez A, Héctor F.; Martínez-Tomás, Rafael; Arias Tapia, Susana A.; Rincón Zamorano, Mariano

2014-04-01

Automatic systems that monitor human behaviour for detecting security problems are a challenge today. Previously, our group defined the Horus framework, which is a modular architecture for the integration of multi-sensor monitoring stages. In this work, structure and technologies required for high-level semantic stages of Horus are proposed, and the associated methodological principles established with the aim of recognising specific behaviours and situations. Our methodology distinguishes three semantic levels of events: low level (compromised with sensors), medium level (compromised with context), and high level (target behaviours). The ontology for surveillance and ubiquitous computing has been used to integrate ontologies from specific domains and together with semantic technologies have facilitated the modelling and implementation of scenes and situations by reusing components. A home context and a supermarket context were modelled following this approach, where three suspicious activities were monitored via different virtual sensors. The experiments demonstrate that our proposals facilitate the rapid prototyping of this kind of systems.

[Antimicrobial activity of Laetiporus sulphureus strains grown in submerged culture].

PubMed

Ershova, E Iu; Tikhonova, O V; Lur'e, L M; Efremenkova, O V; Kamzolkina, O V; Dudnik, Iu V

2003-01-01

Cultural conditions for growth and fruit body formation were elaborated to four strains of Laetiporus sulphureus isolated from nature. All strains demonstrated antimicrobial activity against a wide spectrum of gram-positive and gram-negative bacteria during agar and submerged cultivation including methicillin-resistant strain of Staphylococcus aureus (MRSA) and glycopeptide-resistant strain of Leuconostoc mesenteroides. Antifungal activity was not found. The level of antimicrobial activity during submerged cultivation reached maximum after seven days of growth on specific medium with soybean meal and corn liquid; the next four weeks its increasing was not so manifested. Antimicrobial activity correlated with orange pigment secretion and cultural liquid acidification to pH 2.0-2.8 that indicates on acid nature of synthesized products.

Extract of medicinal mushroom Agaricus blazei Murill enhances the non-specific and adaptive immune activities in BALB/c mice.

PubMed

Ni, Wei-Ya; Wu, Ming-Fanf; Liao, Nien-Chieh; Yeh, Ming-Yang; Lu, Hsu-Feng; Hsueh, Shu-Ching; Liu, Jia-You; Huang, Yi-Ping; Chang, Chuan-Hsun; Chung, Jing-Gung

2013-01-01

Agaricus blazei Murill (AbM) is traditionally used against a wide range of conditions such as ulcerative colitis, Crohn's disease, foot-and-mouth disease and chronic hepatitis C infection. In this study, we evaluated the immunomodulatory effects of AbM. For the non-specific immune response experiments, a total of 40 female BALB/c mice were divided into control (group 1) and experimental (groups 2-4) groups of 10 animals each. Groups 2, 3 and 4 were orally-administered high (819 mg/kg), medium (273 mg/kg) and low (136.5 mg/kg) doses of AbM daily for six weeks and then six parameters related to non-specific immune response were detected. For the adaptive immune response experiments, 40 female mice were similarly divided into four groups. After six weeks of treatment, animals were immunized with the OVA immunogen. Two weeks later, splenocytes and sera were collected. Four parameters related to adaptive immune response were evaluated. We found that feeding mice with AbM extract increased the IgG level in serum, promoted phagocytosis of peritoneal macrophages and elevated the activity of Natural killer cells. We also found that the highest dose of AbM increased interleukin-2 (IL-2) levels in splenocytes and that a medium dose increased interferon-γ. The levels of interleukin-4 (IL-4) were reduced or unchanged. T-helper type 1 cytokine levels were increased. AbM increased the humoral immune response and also affected the cellular immune response. These results provide evidence that AbM can modulate innate and adaptive immunity.

Single ingestion of soy β-conglycinin induces increased postprandial circulating FGF21 levels exerting beneficial health effects.

PubMed

Hashidume, Tsutomu; Kato, Asuka; Tanaka, Tomohiro; Miyoshi, Shoko; Itoh, Nobuyuki; Nakata, Rieko; Inoue, Hiroyasu; Oikawa, Akira; Nakai, Yuji; Shimizu, Makoto; Inoue, Jun; Sato, Ryuichiro

2016-06-17

Soy protein β-conglycinin has serum lipid-lowering and anti-obesity effects. We showed that single ingestion of β-conglycinin after fasting alters gene expression in mouse liver. A sharp increase in fibroblast growth factor 21 (FGF21) gene expression, which is depressed by normal feeding, resulted in increased postprandial circulating FGF21 levels along with a significant decrease in adipose tissue weights. Most increases in gene expressions, including FGF21, were targets for the activating transcription factor 4 (ATF4), but not for peroxisome proliferator-activated receptor α. Overexpression of a dominant-negative form of ATF4 significantly reduced β-conglycinin-induced increases in hepatic FGF21 gene expression. In FGF21-deficient mice, β-conglycinin effects were partially abolished. Methionine supplementation to the diet or primary hepatocyte culture medium demonstrated its importance for activating liver or hepatocyte ATF4-FGF21 signaling. Thus, dietary β-conglycinin intake can impact hepatic and systemic metabolism by increasing the postprandial circulating FGF21 levels.

Sinensetin enhances adipogenesis and lipolysis by increasing cyclic adenosine monophosphate levels in 3T3-L1 adipocytes.

PubMed

Kang, Seong-Il; Shin, Hye-Sun; Kim, Se-Jae

2015-01-01

Sinensetin is a rare polymethoxylated flavone (PMF) found in certain citrus fruits. In this study, we investigated the effects of sinensetin on lipid metabolism in 3T3-L1 cells. Sinensetin promoted adipogenesis in 3T3-L1 preadipocytes growing in incomplete differentiation medium, which did not contain 3-isobutyl-1-methylxanthine. Sinensetin up-regulated expression of the adipogenic transcription factors peroxisome proliferator-activated receptor γ, CCAAT/enhancer-binding protein (C/EBP) α, and sterol regulatory element-binding protein 1c. It also potentiated expression of C/EBPβ and activation of cAMP-responsive element-binding protein. Sinensetin enhanced activation of protein kinase A and increased intracellular cAMP levels in 3T3-L1 preadipocytes. In mature 3T3-L1 adipocytes, sinensetin stimulated lipolysis via a cAMP pathway. Taken together, these results suggest that sinensetin enhances adipogenesis and lipolysis by increasing cAMP levels in adipocytes.

Effects of SNF1 on Maltose Metabolism and Leavening Ability of Baker's Yeast in Lean Dough.

PubMed

Zhang, Cui-Ying; Bai, Xiao-Wen; Lin, Xue; Liu, Xiao-Er; Xiao, Dong-Guang

2015-12-01

Maltose metabolism of baker's yeast (Saccharomyces cerevisiae) in lean dough is negatively influenced by glucose repression, thereby delaying the dough fermentation. To improve maltose metabolism and leavening ability, it is necessary to alleviate glucose repression. The Snf1 protein kinase is well known to be essential for the response to glucose repression and required for transcription of glucose-repressed genes including the maltose-utilization genes (MAL). In this study, the SNF1 overexpression and deletion industrial baker's yeast strains were constructed and characterized in terms of maltose utilization, growth and fermentation characteristics, mRNA levels of MAL genes (MAL62 encoding the maltase and MAL61 encoding the maltose permease) and maltase and maltose permease activities. Our results suggest that overexpression of SNF1 was effective to glucose derepression for enhancing MAL expression levels and enzymes (maltase and maltose permease) activities. These enhancements could result in an 18% increase in maltose metabolism of industrial baker's yeast in LSMLD medium (the low sugar model liquid dough fermentation medium) containing glucose and maltose and a 15% increase in leavening ability in lean dough. These findings provide a valuable insight of breeding industrial baker's yeast for rapid fermentation. © 2015 Institute of Food Technologists®

Purine biosynthesis is the bottleneck in trimethoprim-treated Bacillus subtilis.

PubMed

Stepanek, Jennifer Janina; Schäkermann, Sina; Wenzel, Michaela; Prochnow, Pascal; Bandow, Julia Elisabeth

2016-10-01

Trimethoprim is a folate biosynthesis inhibitor. Tetrahydrofolates are essential for the transfer of C 1 units in several biochemical pathways including purine, thymine, methionine, and glycine biosynthesis. This study addressed the effects of folate biosynthesis inhibition on bacterial physiology. Two complementary proteomic approaches were employed to analyze the response of Bacillus subtilis to trimethoprim. Acute changes in protein synthesis rates were monitored by radioactive pulse labeling of newly synthesized proteins and subsequent 2DE analysis. Changes in protein levels were detected using gel-free quantitative MS. Proteins involved in purine and histidine biosynthesis, the σ B -dependent general stress response, and sporulation were upregulated. Most prominently, the PurR-regulon required for de novo purine biosynthesis was derepressed indicating purine depletion. The general stress response was activated energy dependently and in a subpopulation of treated cultures an early onset of sporulation was observed, most likely triggered by low guanosine triphosphate levels. Supplementation of adenosine triphosphate, adenosine, and guanosine to the medium substantially decreased antibacterial activity, showing that purine depletion becomes the bottleneck in trimethoprim-treated B. subtilis. The frequently prescribed antibiotic trimethoprim causes purine depletion in B. subtilis, which can be complemented by supplementing purines to the medium. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Notable fibrolytic enzyme production by Aspergillus spp. isolates from the gastrointestinal tract of beef cattle fed in lignified pastures

PubMed Central

Abrão, Flávia Oliveira; Pessoa, Moisés Sena; dos Santos, Vera Lúcia; de Freitas Júnior, Luiz Fernando; Barros, Katharina de Oliveira; Hughes, Alice Ferreira da Silva; Silva, Thiago Dias; Rodriguez, Norberto Mário

2017-01-01

Fungi have the ability to degrade vegetal cell wall carbohydrates, and their presence in the digestive tract of ruminants can minimize the effects of lignified forage on ruminal fermentation. Here, we evaluated enzyme production by Aspergillus spp. isolates from the digestive tracts of cattle grazed in tropical pastures during the dry season. Filamentous fungi were isolated from rumen and feces by culture in cellulose-based medium. Ninety fungal strains were isolated and identified by rDNA sequence analysis, microculture, or both. Aspergillus terreus was the most frequently isolated species, followed by Aspergillus fumigatus. The isolates were characterized with respect to their cellulolytic, xylanolytic, and lignolytic activity through qualitative evaluation in culture medium containing a specific corresponding carbon source. Carboxymethyl cellulase (CMCase) activity was quantified by the reducing sugar method. In the avicel and xilan degradation test, the enzyme activity (EA) at 48 h was significantly higher other periods (P < 0.05). Intra- and inter-specific differences in EA were verified, and high levels of phenoloxidases, which are crucial for lignin degradation, were observed in 28.9% of the isolates. Aspergillus terreus showed significantly higher EA for avicelase (3.96 ±1.77) and xylanase (3.13 ±.091) than the other Aspergillus species at 48 h of incubation. Isolates AT13 and AF69 showed the highest CMCase specific activity (54.84 and 33.03 U mg-1 protein, respectively). Selected Aspergillus spp. isolates produced remarkable levels of enzymes involved in vegetal cell wall degradation, suggesting their potential as antimicrobial additives or probiotics in ruminant diets. PMID:28850605

The effect of ethylenediaminetetraacetic acid on heat resistance and recovery of Clostridium sporogenes PA 3679 spores treated in HTST conditions.

PubMed

Silla Santos, M H; Torres Zarzo, J

1997-03-03

The effect of ethylenediaminetetraacetic acid (EDTA) on the heat resistance of Clostridium sporogenes PA 3679 spores was studied. EDTA was added to heating substrates and recovery media in order to establish which stage of the heat treatment registered the greatest EDTA activity. The heating substrates assayed were phosphate buffer (pH 7.0) and white asparagus purée, at natural pH (5.8) and acidified with citric acid and glucono-delta-lactone (GDL) to pH 5.5, 5.0 and 4.5. Recovery of survivors was carried out in MPA3679A medium in various conditions of acidification with citric and GDL (250 and 500 ppm), at pH 7.5 6.5 and 6.0. The results show greater activity of EDTA on spores when it was applied in recovery of heat injured spores, than during heating. The strongest influence of EDTA during heating was found in phosphate buffer (pH 7.0), with the effect being most evident at 121 and 126 degrees C, and in asparagus purée, at 121 degrees C and pH 5.8 rather than acidified. In recovery, the inhibiting activity of EDTA was more evident in spores subjected to more severe heat treatment, either by increasing the exposure time or by raising the temperature to 130 or 135 degrees C. The pH level of the recovery medium also affected the antimicrobial activity of EDTA, which had a greater inhibiting effect at pH 7.5 than at lower pH levels (6.5, 6.0).

Notable fibrolytic enzyme production by Aspergillus spp. isolates from the gastrointestinal tract of beef cattle fed in lignified pastures.

PubMed

Abrão, Flávia Oliveira; Duarte, Eduardo Robson; Pessoa, Moisés Sena; Santos, Vera Lúcia Dos; Freitas Júnior, Luiz Fernando de; Barros, Katharina de Oliveira; Hughes, Alice Ferreira da Silva; Silva, Thiago Dias; Rodriguez, Norberto Mário

2017-01-01

Fungi have the ability to degrade vegetal cell wall carbohydrates, and their presence in the digestive tract of ruminants can minimize the effects of lignified forage on ruminal fermentation. Here, we evaluated enzyme production by Aspergillus spp. isolates from the digestive tracts of cattle grazed in tropical pastures during the dry season. Filamentous fungi were isolated from rumen and feces by culture in cellulose-based medium. Ninety fungal strains were isolated and identified by rDNA sequence analysis, microculture, or both. Aspergillus terreus was the most frequently isolated species, followed by Aspergillus fumigatus. The isolates were characterized with respect to their cellulolytic, xylanolytic, and lignolytic activity through qualitative evaluation in culture medium containing a specific corresponding carbon source. Carboxymethyl cellulase (CMCase) activity was quantified by the reducing sugar method. In the avicel and xilan degradation test, the enzyme activity (EA) at 48 h was significantly higher other periods (P < 0.05). Intra- and inter-specific differences in EA were verified, and high levels of phenoloxidases, which are crucial for lignin degradation, were observed in 28.9% of the isolates. Aspergillus terreus showed significantly higher EA for avicelase (3.96 ±1.77) and xylanase (3.13 ±.091) than the other Aspergillus species at 48 h of incubation. Isolates AT13 and AF69 showed the highest CMCase specific activity (54.84 and 33.03 U mg-1 protein, respectively). Selected Aspergillus spp. isolates produced remarkable levels of enzymes involved in vegetal cell wall degradation, suggesting their potential as antimicrobial additives or probiotics in ruminant diets.

Inhibition of biosynthesis of metalloprotease of Aeromonas sobria by sodium chloride in the medium.

PubMed

Takahashi, Eizo; Kobayashi, Hidetomo; Yamanaka, Hiroyasu; Nair, Gopinath Balakrish; Takeda, Yoshifumi; Arimoto, Sakae; Negishi, Tomoe; Okamoto, Keinosuke

2011-01-01

The present authors have previously shown that the serine protease activity of Aeromonas sobria is markedly decreased when A. sobria is cultured in medium containing 3.0% sodium chloride (NaCl, concentration almost equivalent to sea water salinity), and that this occurs because, although the synthesis of ASP is not disturbed by the salt in the medium, the maturation pathway of serine protease of A. sobria (ASP) does not proceed successfully in such a medium. In this study, the effect of salt in the medium on the production of metalloprotease by A. sobria (AMP) was examined. A. sobria produced AMP in the milieu when the bacteria were cultured in medium containing (NaCl) at a concentration of 0.5%. However, AMP was not produced when the bacteria were cultured in salty medium containing 1.5% or more NaCl. To examine how NaCl reduces the production of metalloprotease by A. sobria, the amount of amp mRNA in the cell was measured and it was found that this decreased in proportion to the concentration of NaCl in the medium. The mRNA of amp was not detected in cells cultured in medium containing 1.5% or more NaCl. This means that the transcription of amp is inhibited in salty condition. As described, NaCl in the medium disturbs the maturation pathway of ASP. The mode of action whereby NaCl suppresses AMP activity in A. sobria differs from the mechanism for suppressing ASP activity. © 2010 The Societies and Blackwell Publishing Asia Pty Ltd.

Relative quantification of beta-casein expression in primary goat mammary epithelial cell lines.

PubMed

Ogorevc, J; Dovč, P

2015-04-15

Primary mammary epithelial cell cultures were established from mammary tissue of lactating and non-lactating goats to assess the expression of beta-casein (CSN2) in vitro. Primary cell cultures were established by enzymatic digestion of mammary tissue and characterized using antibodies against cytokeratin 14, cytokeratin 18, and vimentin. The established primary cell lines in the second passage were grown in basal medium on plastic and in hormone-supplemented (lactogenic) medium on plastic and on an extracellular matrix-covered surface, respectively. CSN2 gene expression was evaluated using quantitative reverse transcription PCR. The presence of CSN2 transcripts was detected in all samples, including cells originating from non-lactating goat, grown in basal medium. The presence of CSN2 protein was confirmed using immunofluorescence. Response to the hormonal treatment and cell morphology differed between the cell lines and treatments. In 2 cell lines supplemented with lactogenic hormones in the medium, CSN2 expression was increased, while CSN2 levels in one of the cell lines remained constant, regardless of the treatment. Addition of extracellular matrix showed positive effects on CSN2 transcription activity in 1 of the cell lines, while in the other 2 showed no statistically significant effects. CSN2 expression appeared to depend on subtle differences in physiological state of the starting tissue material, growth conditions, cell types present in the culture, and methods used for cell culture establishment. Further studies are necessary to identify factors that determine hormone-responsiveness and transcriptional activity of milk protein genes in goat primary mammary cell cultures.

Frequency Tunable Wire Lasers

NASA Technical Reports Server (NTRS)

Hu, Qing (Inventor)

2013-01-01

The present invention provides frequency tunable solid-state radiation-generating devices, such as lasers and amplifiers, whose active medium has a size in at least one transverse dimension (e.g., its width) that is much smaller than the wavelength of radiation generated and/or amplified within the active medium. In such devices, a fraction of radiation travels as an evanescent propagating mode outside the active medium. It has been discovered that in such devices the radiation frequency can be tuned by the interaction of a tuning mechanism with the propagating evanescent mode.

In vitro transdentinal effect of low-level laser therapy

NASA Astrophysics Data System (ADS)

Oliveira, C. F.; Basso, F. G.; dos Reis, R. I.; Parreiras-e-Silva, L. T.; Lins, E. C.; Kurachi, C.; Hebling, J.; Bagnato, V. S.; de Souza Costa, C. A.

2013-05-01

Low-level laser therapy (LLLT) has been used for the treatment of dentinal hypersensitivity. However, the specific LLL dose and the response mechanisms of these cells to transdentinal irradiation have not yet been demonstrated. Therefore, this study evaluated the transdentinal effects of different LLL doses on stressed odontoblast-like pulp cells MDPC-23 seeded onto the pulpal side of dentin discs obtained from human third molars. The discs were placed in devices simulating in vitro pulp chambers and the whole set was placed in 24-well plates containing plain culture medium (DMEM). After 24 h incubation, the culture medium was replaced by fresh DMEM supplemented with either 5% (simulating a nutritional stress condition) or 10% fetal bovine serum (FBS). The cells were irradiated with doses of 15 and 25 J cm-2 every 24 h, totaling three applications over three consecutive days. The cells in the control groups were removed from the incubator for the same times as used in their respective experimental groups for irradiation, though without activating the laser source (sham irradiation). After 72 h of the last active or sham irradiation, the cells were evaluated with respect to succinic dehydrogenase (SDH) enzyme production (MTT assay), total protein (TP) expression, alkaline phosphatase (ALP) synthesis, reverse transcriptase polymerase chain reaction (RT-PCR) for collagen type 1 (Col-I) and ALP, and morphology (SEM). For both tests, significantly higher values were obtained for the 25 J cm-2 dose. Regarding SDH production, supplementation of the culture medium with 5% FBS provided better results. For TP and ALP expression, the 25 J cm-2 presented higher values, especially for the 5% FBS concentration (Mann-Whitney p < 0.05). Under the tested conditions, near infrared laser irradiation at 25 J cm-2 caused transdentinal biostimulation of odontoblast-like MDPC-23 cells.

Tissue-specific regulation of medium-chain acyl-CoA dehydrogenase gene by thyroid hormones in the developing rat.

PubMed

Djouadi, F; Riveau, B; Merlet-Benichou, C; Bastin, J

1997-05-15

During development, gene expression of medium-chain acyl-CoA dehydrogenase (MCAD), a nuclear-encoded mitochondrial enzyme that catalyses the first step of medium-chain fatty acid beta-oxidation, is highly regulated in tissues in accordance with fatty acid utilization, but the factors involved in this regulation are largely unknown. To investigate a possible role of thyroid hormones, rat pups were made hypothyroid by the administration of propylthiouracyl to the mother from day 12 of gestation, and their kidneys, heart and liver were removed on postnatal day 16 to determine MCAD mRNA abundance, protein level and enzyme activity. Similar experiments were run in 3,3',5-tri-iodothyronine (T3)-replaced hypothyroid (1 microg of T3/100 g body weight from postnatal day 5 to 15) and euthyroid pups. Hypothyroidism led to an increase in MCAD mRNA abundance in kidney and a decrease in abundance in heart, but had no effect in liver. The protein levels and enzyme activity were lowered in hypothyroid heart and kidney, suggesting that hypothyroidism affects post-transcriptional steps of gene expression in the kidney. All the effects of hypothyroidism were completely reversed in both heart and kidney by T3 replacement. Injection of a single T3 dose into 16-day-old euthyroid rats also led to tissue-specific changes in mRNA abundance. Nuclear run-on assays performed from hypothyroid and hypothyroid plus T3 rats showed that T3 stimulates MCAD gene transcription in heart and represses it in the kidney. These results indicate that the postnatal rise in circulating T3 is essential to the developmental regulation of the MCAD gene in vivo.

Promotive Effect of Zinc Ions on the Vitality, Migration, and Osteogenic Differentiation of Human Dental Pulp Cells.

PubMed

An, Shaofeng; Gong, Qimei; Huang, Yihua

2017-01-01

Zinc is an essential trace element for proper cellular function and bone formation. However, its exact role in the osteogenic differentiation of human dental pulp cells (hDPCs) has not been fully clarified before. Here, we speculated that zinc may be effective to regulate their growth and osteogenic differentiation properties. To test this hypothesis, different concentrations (1 × 10 -5 , 4 × 10 -5 , and 8 × 10 -5  M) of zinc ions (Zn 2+ ) were added to the basic growth culture medium and osteogenic inductive medium. Cell viability and migration were measured by cell counting kit-8 (CCK-8) and transwell migration assay in the basic growth culture medium, respectively. The reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to detect the gene expression levels of selective osteogenic differentiation markers and zinc transporters. Alkaline phosphatase (ALP) activity analysis and alizarin red S staining were used to investigate the mineralization of hDPCs. Exposure of hDPCs to Zn 2+ stimulated their viability and migration capacity in a dose- and time-dependent manner. RT-qPCR assay revealed elevated expression levels of osteogenic differentiation-related genes and zinc transporters genes in various degrees. ALP activity was also increased with elevated Zn 2+ concentrations and extended culture periods, but enhanced matrix nodules formation were observed only in 4 × 10 -5 and 8 × 10 -5  M Zn 2+ groups. These findings suggest that specific concentrations of Zn 2+ could potentiate the vitality, migration, and osteogenic differentiation of hDPCs. We may combine optimum zinc element into pulp capping materials to improve their biological performance.

Control of 5-aminolaevulinate synthetase activity in Rhodopseudomonas spheroides. The involvement of sulphur metabolism

PubMed Central

Neuberger, Albert; Sandy, John D.; Tait, George H.

1973-01-01

1. The `initial' 5-aminolaevulinate synthetase activity, that is the activity observed immediately after cell disruption, in extracts prepared from unharvested semianaerobically grown Rhodopseudomonas spheroides, was twice that observed under the same assay conditions in extracts prepared from harvested cells. 2. The effect of oxygenation of a culture on the `maximum' aminolaevulinate synthetase activity, that is the activity observed 1h after disruption of harvested cells, is markedly influenced by the contents of the growth medium. Oxygenation of organisms for 1h in the medium in which they have grown produces an 80–90% decrease in maximum activity, whereas similar treatment of organisms resuspended in fresh medium produces less than a 40% decrease. 3. This protective effect of fresh medium is absolutely dependent on the presence of sulphate. When cells are suspended in sulphate-deficient fresh medium, the maximum activity falls by 65–75% even without oxygenation. A high maximum activity is regenerated when sulphate is resupplied. 4. When organisms are oxygenated in the medium in which they have grown, the cellular contents of GSH+GSSG and cysteine+cystine fall very markedly and homolanthionine is formed. Both the fall in aminolaevulinate synthetase activity and the changes in sulphur metabolism are largely prevented by the addition of compounds which stimulate synthesis of cysteine de novo or inhibit the conversion of cysteine S into homocysteine S. 5. The maximum aminolaevulinate synthetase activity was directly proportional to the GSH+GSSG content of all cell preparations. In glutathione-depleted extracts the `low'-activity enzyme could be re-activated in vitro by the addition of GSH, GSSG, cysteine or cystine, whereas in extracts with a high glutathione content the `high'-activity enzyme was unaffected by these sulphur compounds. 6. The activation of low-activity enzyme with exogenous sulphur compounds was prevented by excluding air or by adding NADH. Studies with purified enzyme indicate that sulphur compounds do not interact directly with the enzyme, but that their effect is mediated by a number of other endogenous factors. PMID:4544404

Physiological and Genetic Analyses Leading to Identification of a Biochemical Role for the moeA (Molybdate Metabolism) Gene Product in Escherichia coli†

PubMed Central

Hasona, Adnan; Ray, Ramesh M.; Shanmugam, K. T.

1998-01-01

A unique class of chlorate-resistant mutants of Escherichia coli which produced formate hydrogenlyase and nitrate reductase activities only when grown in medium with limiting amounts of sulfur compounds was isolated. These mutants failed to produce the two molybdoenzyme activities when cultured in rich medium or glucose-minimal medium. The mutations in these mutants were localized in the moeA gene. Mutant strains with polar mutations in moeA which are also moeB did not produce active molybdoenzymes in any of the media tested. moeA mutants with a second mutation in either cysDNCJI or cysH gene lost the ability to produce active molybdoenzyme even when grown in medium limiting in sulfur compounds. The CysDNCJIH proteins along with CysG catalyze the conversion of sulfate to sulfide. Addition of sulfide to the growth medium of moeA cys double mutants suppressed the MoeA− phenotype. These results suggest that in the absence of MoeA protein, the sulfide produced by the sulfate activation/reduction pathway combines with molybdate in the production of activated molybdenum. Since hydrogen sulfide is known to interact with molybdate in the production of thiomolybdate, it is possible that the MoeA-catalyzed activated molybdenum is a form of thiomolybdenum species which is used in the synthesis of molybdenum cofactor from Mo-free molybdopterin. PMID:9515915

Elasticity-induced force reversal between active spinning particles in dense passive media

PubMed Central

Aragones, J. L.; Steimel, J. P.; Alexander-Katz, A.

2016-01-01

The self-organization of active particles is governed by their dynamic effective interactions. Such interactions are controlled by the medium in which such active agents reside. Here we study the interactions between active agents in a dense non-active medium. Our system consists of actuated, spinning, active particles embedded in a dense monolayer of passive, or non-active, particles. We demonstrate that the presence of the passive monolayer alters markedly the properties of the system and results in a reversal of the forces between active spinning particles from repulsive to attractive. The origin of such reversal is due to the coupling between the active stresses and elasticity of the system. This discovery provides a mechanism for the interaction between active agents in complex and structured media, opening up opportunities to tune the interaction range and directionality via the mechanical properties of the medium. PMID:27112961

Functional comparison of the reverse mode of Na+/Ca2+ exchangers NCX1.1 and NCX1.5 expressed in CHO cells.

PubMed

Long, Yan; Wang, Wei-ping; Yuan, Hui; Ma, Shi-ping; Feng, Nan; Wang, Ling; Wang, Xiao-liang

2013-05-01

To investigate the reverse mode function of Na(+)/Ca(2+) exchangers NCX1.1 and NCX1.5 expressed in CHO cells as well as their modulations by PKC and PKA. CHO-K1 cells were transfected with pcDNA3.1 (+) plasmid carrying cDNA of rat cardiac NCX1.1 and brain NCX1.5. The expression of NCX1.1 and NCX1.5 was examined using Western blot analysis. The intracellular Ca(2+) level ([Ca(2+)]i) was measured using Ca(2+) imaging. Whole-cell NCX currents were recorded using patch-clamp technique. Reverse mode NCX activity was elicited by perfusion with Na(+)-free medium. Ca(2+) paradox was induced by Ca(2+)-free EBSS medium, followed by Ca(2+)-containing solution (1.8 or 3.8 mmol/L CaCl2). The protein levels of NCX1.1 and NCX1.5 expressed in CHO cells had no significant difference. The reverse modes of NCX1.1 and NCX1.5 in CHO cells exhibited a transient increase of [Ca(2+)]i, which was followed by a Ca(2+) level plateau at higher external Ca(2+) concentrations. In contrast, the wild type CHO cells showed a steady increase of [Ca(2+)]i at higher external Ca(2+) concentrations. The PKC activator PMA (0.3-10 μmol/L) and PKA activator 8-Br-cAMP (10-100 μmol/L) significantly enhanced the reverse mode activity of NCX1.1 and NCX1.5 in CHO cells. NCX1.1 was 2.4-fold more sensitive to PKC activation than NCX1.5, whereas the sensitivity of the two NCX isoforms to PKA activation had no difference. Both PKC- and PKA-enhanced NCX reverse mode activities in CHO cells were suppressed by NCX inhibitor KB-R7943 (30 μmol/L). Both NCX1.1 and NCX1.5 are functional in regulating and maintaining stable [Ca(2+)]i in CHO cells and differentially regulated by PKA and PKC. The two NCX isoforms might be useful drug targets for heart and brain protection.

Adaptation to rhizosphere acidification is a necessary prerequisite for wheat (Triticum aestivum L.) seedling resistance to ammonium stress.

PubMed

Wang, Feng; Gao, Jingwen; Tian, Zhongwei; Liu, Yang; Abid, Muhammad; Jiang, Dong; Cao, Weixing; Dai, Tingbo

2016-11-01

Because soil acidification accompanies ammonium (NH 4 + ) stress, the tolerance of higher plants to ammonium is associated with their adaptation to root medium acidification. However, the underlying mechanisms of this adaptation have not been fully elucidated. The objective of this study was thus to elucidate the effect of rhizosphere pH on NH 4 + tolerance in different winter wheat cultivars (Triticum aestivum L.). Hydroponic experiments were carried out on two wheat cultivars: AK58 (an NH 4 + -sensitive cultivar) and XM25 (an NH 4 + -tolerant cultivar). Four pH levels resembling acidified (4.0, 5.0, 6.0 and 7.0) were tested and 5 mM NH 4 + nitrogen (AN) was used as a stress treatment, with 5 mM nitrate nitrogen used as a control. The addition of AN led to a severe reduction in biomass and an increase in free NH 4 + , amino acids, and the activities of glutamine synthetase (GS) and glutamate dehydrogenase (GDH) in the shoots and roots of the two wheat cultivars. Further decreases in growth medium pH led to further increases in free NH 4 + , but decreases in total amino acids and the activities of GS and NADH-dependent glutamate synthase (NADH-GDH). However, there was less of an increase in free NH 4 + and less of a reduction in the activities of GS and NADH-GDH in the cultivar XM25 compared with AK58. In addition, total soluble sugar content and the root-to-shoot soluble sugar ratio were also decreased by AN treatment, except in the shoots of XM25. Decreasing pH resulted in lower root-to-shoot soluble sugar ratios with greater reductions in the AK58 cultivar. These results indicate that wheat growth was inhibited significantly by the addition of NH 4 + combined with low pH. Low medium pH reduced the capacity for nitrogen assimilation and interrupted carbohydrate transport between the shoot and root. The NH 4 + -tolerant cultivar XM25 was better adapted to low rhizosphere pH due to its increased capacity for assimilating NH 4 + efficiently and thereby avoiding toxic levels of intracellular NH 4 + at low medium pH. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

Free testosterone as marker of adaptation to medium-intensive exercise.

PubMed

Shkurnikov, M U; Donnikov, A E; Akimov, E B; Sakharov, D A; Tonevitsky, A G

2008-09-01

A 4-week study of adaptation reserves of the body was carried out during medium intensive exercise (medium intensive training: 60-80% threshold anaerobic metabolism). Two groups of athletes were singled out by the results of pulsometry analysis: with less than 20% work duration at the level above the 80% threshold anaerobic metabolism and with more than 20% work duration at the level above 80% threshold anaerobic metabolism. No appreciable differences between the concentrations of total testosterone, growth hormone, and cortisol before and after exercise in the groups with different percentage of anaerobic work duration were detected. In group 1 the concentrations of free testosterone did not change throughout the period of observation in comparison with the levels before training. In group 2, the level of free testosterone increased in comparison with the basal level: from 0.61+/-0.12 nmol/liter at the end of week 1 to 0.98+/-0.11 nmol/liter at the end of week 4 (p<0.01). The results indicate that the level of free testosterone can be used for evaluating the degree of athlete's adaptation to medium intensive exercise.

Heterogeneity in extracellular nucleotide hydrolysis among clinical isolates of Trichomonas vaginalis

PubMed Central

TASCA, T.; BONAN, C. D.; DE CARLI, G. A.; SARKIS, J.J.F.; ALDERETE, J. F.

2007-01-01

SUMMARY Trichomonas vaginalis is a parasitic protozoan that causes trichomonosis, a sexually-transmitted disease, with serious sequelae to women and men. As the host-parasite relationship is complex, it is important to investigate biochemical aspects of the parasite that contribute to our understanding of trichomonal biology and pathogenesis. Nucleoside triphosphate diphosphohydrolase 1 (NTPDase 1), which hydrolyses extracellular ATP and ADP, and ecto-5′-nucleotidase, which hyrolyses AMP, have been characterized in laboratory isolates of T. vaginalis. Here we show that the extracellular ATP:ADP hydrolysis ratio varies among fresh clinical isolates, which presented higher ATPase and ADPase activities than long-term-grown isolates. Growth of parasites in iron-replete and iron-depleted medium resulted in different, albeit minor, patterns in extracellular ATP and ADP hydrolysis among isolates. Importantly, some isolates had low or absent ecto-5′-nucleotidase activity, regardless of environmental conditions tested. For isolates with ecto-5′-nucleotidase activity, high- and low-iron trichomonads had increased and decreased levels of activity, respectively, compared to organisms grown in normal TYM-serum medium. This suggests a regulation in expression of either the enzyme amounts and/or activity under the control of iron. Finally, we found no correlation between the presence or absence of dsRNA virus infection among trichomonad isolates and NTPDase and ecto-5′-nucleotidase activities. PMID:16038398

An aspartic proteinase gene family in the filamentous fungus Botrytis cinerea contains members with novel features.

PubMed

ten Have, Arjen; Dekkers, Ester; Kay, John; Phylip, Lowri H; van Kan, Jan A L

2004-07-01

Botrytis cinerea, an important fungal plant pathogen, secretes aspartic proteinase (AP) activity in axenic cultures. No cysteine, serine or metalloproteinase activity could be detected. Proteinase activity was higher in culture medium containing BSA or wheat germ extract, as compared to minimal medium. A proportion of the enzyme activity remained in the extracellular glucan sheath. AP was also the only type of proteinase activity in fluid obtained from B. cinerea-infected tissue of apple, pepper, tomato and zucchini. Five B. cinerea genes encoding an AP were cloned and denoted Bcap1-5. Features of the encoded proteins are discussed. BcAP1, especially, has novel characteristics. A phylogenetic analysis was performed comprising sequences originating from different kingdoms. BcAP1 and BcAP5 did not cluster in a bootstrap-supported clade. BcAP2 clusters with vacuolar APs. BcAP3 and BcAP4 cluster with secreted APs in a clade that also contains glycosylphosphatidylinositol-anchored proteinases from Saccharomyces cerevisiae and Candida albicans. All five Bcap genes are expressed in liquid cultures. Transcript levels of Bcap1, Bcap2, Bcap3 and Bcap4 are subject to glucose and peptone repression. Transcripts from all five Bcap genes were detected in infected plant tissue, indicating that at least part of the AP activity in planta originates from the pathogen.

The effect of different physical activity levels on muscle fiber size and type distribution of lumbar multifidus. A biopsy study on low back pain patient groups and healthy control subjects.

PubMed

Mazis, N; Papachristou, D J; Zouboulis, P; Tyllianakis, M; Scopa, C D; Megas, P

2009-12-01

Previous studies examining the multifidus fiber characteristics among low back pain (LBP) patients have not considered the variable of physical activity. The present study sought to investigate the muscle fiber size and type distribution of the lumbar multifidus muscle among LBP patient groups with different physical activity levels and healthy controls. Sixty-four patients were assigned to one of three groups named according to the physical activity level, determined for each patient by the International Physical Activity Questionnaire. These were low (LPA), medium (MPA) and high (HPA) physical activity groups. A control group comprising of 17 healthy individuals was also recruited. Muscle biopsy samples were obtained from the multifidus muscle at the level L4-L5. contrast with the control group, LBP patient groups showed a significantly higher Type II fiber distribution as well as reduced diameter in both fiber types (P<0.05). The physical activity level did not have an effect on multifidus characteristics since no significant differences were observed in fiber type and diameter (P>0.05) among LPA, MPA and HPA patient groups. Various pathological conditions were detected which were more pronounced in LBP groups compared to the control (P<0.05). Males had a larger fiber diameter compared to females for both fiber types (P<0.05). The results showed that the level of physical activity did not affect muscle fiber size and type distribution among LBP patients groups. These findings suggest that not only inactivity but also high physical activity levels can have an adverse effect on the multifidus muscle fiber characteristics.

Evaluation of the effects of a plasma activated medium on cancer cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mohades, S.; Laroussi, M., E-mail: mlarouss@odu.edu; Sears, J.

2015-12-15

The interaction of low temperature plasma with liquids is a relevant topic of study to the field of plasma medicine. This is because cells and tissues are normally surrounded or covered by biological fluids. Therefore, the chemistry induced by the plasma in the aqueous state becomes crucial and usually dictates the biological outcomes. This process became even more important after the discovery that plasma activated media can be useful in killing various cancer cell lines. Here, we report on the measurements of concentrations of hydrogen peroxide, a species known to have strong biological effects, produced by application of plasma tomore » a minimum essential culture medium. The activated medium is then used to treat SCaBER cancer cells. Results indicate that the plasma activated medium can kill the cancer cells in a dose dependent manner, retain its killing effect for several hours, and is as effective as apoptosis inducing drugs.« less

LASERS: Efficient chemical oxygen — iodine laser with a high total pressure of the active medium

NASA Astrophysics Data System (ADS)

Zagidullin, M. V.; Nikolaev, V. D.; Svistun, M. I.; Khvatov, N. A.; Heiger, G. D.; Madden, T. J.

2001-01-01

A new concept of obtaining a high total pressure of the active medium of a chemical oxygen — iodine laser (OIL) is proposed and verified. The nozzle unit of the laser consists of the alternating vertical arrays of cylindrical nozzles to produce high-pressure nitrogen jets, plane slotted nozzles for the flow of O2(1Δ) oxygen, and vertical arrays of cylindrical nozzles to inject the N2 — I2 mixture between the first two streams. For a molar chlorine flow rate of 39.2 mmol s-1, the output power was 700 W and the chemical efficiency was 19.7 %. The combined use of the ejector nozzle unit proposed to obtain the active medium and a super-sonic diffuser allows a significant simplification of the ejection system for the exhaust active medium of the OIL.

Signal Factors Secreted by 2D and Spheroid Mesenchymal Stem Cells and by Cocultures of Mesenchymal Stem Cells Derived Microvesicles and Retinal Photoreceptor Neurons

PubMed Central

Mao, Mao; Zhou, Liang

2017-01-01

We aim to identify levels of signal factors secreted by MSCs cultured in 2D monolayers (2D-MSCs), spheroids (spheroids MSCs), and cocultures of microvesicles (MVs) derived from 2D-MSCs or spheroid MSCs and retinal photoreceptor neurons. We seeded 2D-MSCs, spheroid MSCs, and cells derived from spheroids MSCs at equal numbers. MVs isolated from all 3 culture conditions were incubated with 661W cells. Levels of 51 signal factors in conditioned medium from those cultured conditions were quantified with bead-based assay. We found that IL-8, IL-6, and GROα were the top three most abundant signal factors. Moreover, compared to 2D-MSCs, levels of 11 cytokines and IL-2Rα were significantly increased in conditioned medium from spheroid MSCs. Finally, to test if enhanced expression of these factors reflects altered immunomodulating activities, we assessed the effect of 2D-MSC-MVs and 3D-MSC-MVs on CD14+ cell chemoattraction. Compared to 2D-MSC-MVs, 3D-MSC-MVs significantly decreased the chemotactic index of CD14+ cells. Our results suggest that spheroid culture conditions improve the ability of MSCs to selectively secrete signal factors. Moreover, 3D-MSC-MVs also possessed an enhanced capability to promote signal factors secretion compared to 2D-MSC-MVs and may possess enhanced immunomodulating activities and might be a better regenerative therapy for retinal degenerative diseases. PMID:28194184

Link between lipid metabolism and voluntary food intake in rainbow trout fed coconut oil rich in medium-chain TAG.

PubMed

Figueiredo-Silva, A Cláudia; Kaushik, Sadasivam; Terrier, Frédéric; Schrama, Johan W; Médale, Françoise; Geurden, Inge

2012-06-01

We examined the long-term effect of feeding coconut oil (CO; rich in lauric acid, C12) on voluntary food intake and nutrient utilisation in rainbow trout (Oncorhynchus mykiss), with particular attention to the metabolic use (storage or oxidation) of ingested medium-chain TAG. Trout were fed for 15 weeks one of the four isoproteic diets containing fish oil (FO) or CO as fat source (FS), incorporated at 5% (low fat, LF) or 15% (high fat, HF). Fat level or FS did not modify food intake (g/kg(0·8) per d), despite higher intestinal cholecystokinin-T mRNA in trout fed the HF-FO diet. The HF diets relative to the LF ones induced higher growth and adiposity, whereas the replacements of FO by CO resulted in similar growth and adiposity. This, together with the substantial retention of C12 (57% of intake), suggests the relatively low oxidation of ingested C12. The down-regulation of carnitine palmitoyl-transferase-1 (CPT-1) confirms the minor dependency of medium-chain fatty acids (MCFA) on CPT-1 to enter the mitochondria. However, MCFA did not up-regulate mitochondrial oxidation evaluated using hepatic hydroxyacyl-CoA dehydrogenase as a marker, in line with their high retention in body lipids. At a low lipid level, MCFA increased mRNA levels of fatty acid synthase, elongase and stearoyl-CoA desaturase in liver, showing the hepatic activation of fatty acid synthesis pathways by MCFA, reflected by increased 16 : 0, 18 : 0, 16 : 1, 18 : 1 body levels. The high capacity of trout to incorporate and transform C12, rather than to readily oxidise C12, contrasts with data in mammals and may explain the absence of a satiating effect of CO in rainbow trout.

Defective Response of Natural Killer Activity to Thyroxine in Graves’ Disease

PubMed Central

Lee, Myung-Shik; Hong, Weon-Seon; Hong, Seong Woon; Lee, Jhin-Oh; Kang, Tae-Woong

1990-01-01

The effect of thyroxine (T4) on natural killer (NK) activity of peripheral blood lymphocytes (PBL) was investigated, using a 4-hr 51Cr release assay, in 18 patients with previously untreated Graves’ disease and in 18 controls. NK activity in patients with Graves disease was not significantly different from that in the controls. Normal T4 (NT) and high T4 (HT) medium, free T4 concentrations in which were 1.01 and 16.3 ng/dl, respectively, were used to evaluate the effect of T4 on NK activity. In the controls, NK activity increased in the NT or HT medium compared with that in the control medium at effector to target cell (E : T) ratios of 25 : 1 and 50 : 1. NK activity in the Graves’ disease patients, however, did not increase when either the NT or HT medium was used at E : T ratios of 25 : 1 and 50 : 1. These results suggest that patients with Graves’ disease have a similar NK activity to the controls but have a defect in the peripheral blood lymphocytes to increase NK activity in response to T4. PMID:2098098

Coherent perfect absorption in a homogeneously broadened two-level medium

DOE Office of Scientific and Technical Information (OSTI.GOV)

Longhi, Stefano

2011-05-15

In recent works, it has been shown, rather generally, that the time-reversed process of lasing at threshold realizes a coherent perfect absorber (CPA). In a CPA, a lossy medium in an optical cavity with a specific degree of dissipation, equal in modulus to the gain of the lasing medium, can perfectly absorb coherent optical waves that are the time-reversed counterpart of the lasing field. Here, the time-reversed process of lasing is considered in detail for a homogeneously broadened two-level medium in an optical cavity and the conditions for CPA are derived. It is shown that, owing to the dispersive propertiesmore » of the two-level medium, exact time-reversal symmetry is broken and the frequency of the field at which CPA occurs is generally different than the one of the lasing mode. Moreover, at a large cooperation parameter, the observation of CPA in the presence of bistability requires one to operate in the upper branch of the hysteresis cycle.« less

24 CFR 576.106 - Short-term and medium-term rental assistance.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 24 Housing and Urban Development 3 2012-04-01 2012-04-01 false Short-term and medium-term rental... and Eligible Activities § 576.106 Short-term and medium-term rental assistance. (a) General provisions... assistance may be short-term rental assistance, medium-term rental assistance, payment of rental arrears, or...

24 CFR 576.106 - Short-term and medium-term rental assistance.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 24 Housing and Urban Development 3 2013-04-01 2013-04-01 false Short-term and medium-term rental... and Eligible Activities § 576.106 Short-term and medium-term rental assistance. (a) General provisions... assistance may be short-term rental assistance, medium-term rental assistance, payment of rental arrears, or...

24 CFR 576.106 - Short-term and medium-term rental assistance.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 24 Housing and Urban Development 3 2014-04-01 2013-04-01 true Short-term and medium-term rental... and Eligible Activities § 576.106 Short-term and medium-term rental assistance. (a) General provisions... assistance may be short-term rental assistance, medium-term rental assistance, payment of rental arrears, or...

Pre-harvest UV-C irradiation triggers VOCs accumulation with alteration of antioxidant enzymes and phytohormones in strawberry leaves.

PubMed

Xu, Yanqun; Luo, Zisheng; Charles, Marie Thérèse; Rolland, Daniel; Roussel, Dominique

2017-11-01

Recent studies have highlighted the biological and physiological effects of pre-harvest ultraviolet (UV)-C treatment on growing plants. However, little is known about the involvement of volatile organic compounds (VOCs) and their response to this treatment. In this study, strawberry plants were exposed to three different doses of UV-C radiation for seven weeks (a low dose: 9.6kJm -2 ; a medium dose: 15kJm -2 ; and a high-dose: 29.4kJm -2 ). Changes in VOC profiles were investigated and an attempt was made to identify factors that may be involved in the regulation of these alterations. Principle compounds analysis revealed that VOC profiles of UV-C treated samples were significantly altered with 26 VOCs being the major contributors to segregation. Among them, 18 fatty acid-derived VOCs accumulated in plants that received high and medium dose of UV-C treatments with higher lipoxygenase and alcohol dehydrogenase activities. In treated samples, the activity of the antioxidant enzymes catalase and peroxidase was inhibited, resulting in a reduced antioxidant capacity and higher lipid peroxidation. Simultaneously, jasmonic acid level was 74% higher in the high-dose group while abscisic acid content was more than 12% lower in both the medium and high-dose UV-C treated samples. These results indicated that pre-harvest UV-C treatment stimulated the biosynthesis of fatty acid-derived VOCs in strawberry leaf tissue by upregulating the activity of enzymes of the LOX biosynthetic pathway and downregulating antioxidant enzyme activities. It is further suggested that the mechanisms underlying fatty acid-derived VOCs biosynthesis in UV-C treated strawberry leaves are associated with UV-C-induced changes in phytohormone profiles. Crown Copyright © 2017. Published by Elsevier GmbH. All rights reserved.

Studies on the production and purification of an antimicrobial compound and taxonomy of the producer isolated from the marine environment of the Sundarbans.

PubMed

Saha, M; Ghosh, D; Ghosh, D; Garai, D; Jaisankar, P; Sarkar, K K; Dutta, P K; Das, S; Jha, T; Mukherjee, J

2005-02-01

A microorganism isolated from the Sundarbans region of the Bay of Bengal, India, showed potent antimicrobial activity against gram-positive and gram-negative bacteria, molds, yeast and several multiple-drug-resistant (MDR) bacteria, including methicillin-resistant Staphylococcus aureus (MRSA). The isolate grew in the presence of 20% (w/v) NaCl, antibiotic production being maximum with 5% (w/v) NaCl in the production medium. Natural seawater stimulated antibiotic biosynthesis. The absence of catabolite repression during the synthesis of the antimicrobial substance was demonstrated by the utilization of glucose by this isolate. The 16S rRNA gene of this aerobic, gram-positive, mycelium- and spore-forming microorganism was amplified, and molecular phylogenetic analysis of the DNA sequence showed less than 93% similarity with its closest relative, indicating differentiation at the genus level. The highly stable, active principle was purified by butyl acetate extraction and silica-gel chromatography and a single compound was found to posses the broad-spectrum activity. Molecular characterization showed that the active compound is a lipid. Bioreactor studies demonstrated that antibiotic production is strongly dependent on the scale of operation and there is a definite relation between the dissolved oxygen concentration, medium pH, glucose utilization, cell differentiation and antibiotic production. Maximum production in 30 h could be obtained by regulation of the medium pH in the alkaline range by a combination of controlled addition of NaOH, regulation of the air supply and changes in the reactor configuration. Considering all of the above evidences and based on comparison with the current literature, a novel antimicrobial appears to have been isolated.

Azolla filiculoides Nitrogenase Activity Decrease Induced by Inoculation with Chlamydomonas sp. †

PubMed Central

Habte, Mitiku

1986-01-01

Experiments were conducted to determine the influence of Chlamydomonas sp. on nitrogen fixation (C2H2 → C2H4) in Azolla filiculoides and on the nitrogen fixation and growth of free-living Anabaena azollae 2B organisms. Inoculation of azolla medium with Chlamydomonas sp. was associated with decreased nitrogenase activity in A. filiculoides and with increases in the density of a fungal population identified as Acremonium sp. Subsequent inoculation of azolla medium with this fungus was also accompanied by a significant decrease in nitrogenase activity of A. filiculoides. However, the extent of depression of nitrogenase activity was significantly higher when azolla medium was inoculated with Chlamydomonas sp. than when it was inoculated with Acremonium sp. Inoculation of nitrogen-free Stanier medium with either Acremonium sp. or Chlamydomonas sp. did not adversely affect the growth or nitrogenase activity of free-living A. azollae. Decreased nitrogenase activity in A. filiculoides is apparently related to the adverse influence of the green alga and the fungus on the macrosymbiont. The mechanisms that might be involved are discussed. PMID:16347211

Use of microbial activity measurements for monitoring RBC biofilms.

PubMed

Coello, M Dolores; Rodríguez-Barroso, M R; Aragón, C A; Quiroga, J M

2010-10-01

Fixed biomass technologies, such as rotating biological contactors or biodiscs, have been applied for wastewater depuration both in large and medium-small-sized agglomerations. Biofilm's composition and microorganism activity are essential parameters for the successful operation and control of these systems. Biofilm's thickness and total dry weight have been widely used for biofilm's characterization but, actually, are not sufficient to describe biofilm activity. In fact, biofilm's activity is not proportional to the quantity of fixed biomass, but increases with the thickness of biofilm up to a determined level called the "active thickness". Above this level, the diffusion of nutrients through the film becomes a limiting factor. A stable, thin, and active biofilm thus offers numerous advantages in water and wastewater treatment. Different parameters have been used to evaluate biofilm's activity. The specific oxygen uptake rate, INT-dehydrogenase activity, and the ratio active/total cells have been applied for that purpose. These methods are not only simple and rapid but also sensitive, precise, and representative. The results obtained confirm the potential of the microbial activity measurements studied for an accurate biofilm's characterization and biomass activity estimation in fundamental research and for the practical operation and control of fixed biomass depuration systems.

Semiconductor switch geometry with electric field shaping

DOEpatents

Booth, R.; Pocha, M.D.

1994-08-23

An optoelectric switch is disclosed that utilizes a cylindrically shaped and contoured GaAs medium or other optically active semiconductor medium to couple two cylindrically shaped metal conductors with flat and flared termination points each having an ovoid prominence centrally extending there from. Coupling the truncated ovoid prominence of each conductor with the cylindrically shaped optically active semiconductor causes the semiconductor to cylindrically taper to a triple junction circular line at the base of each prominence where the metal conductor conjoins with the semiconductor and a third medium such as epoxy or air. Tapering the semiconductor at the triple junction inhibits carrier formation and injection at the triple junction and thereby enables greater current carrying capacity through and greater sensitivity of the bulk area of the optically active medium. 10 figs.

Semiconductor switch geometry with electric field shaping

DOEpatents

Booth, Rex; Pocha, Michael D.

1994-01-01

An optoelectric switch is disclosed that utilizes a cylindrically shaped and contoured GaAs medium or other optically active semiconductor medium to couple two cylindrically shaped metal conductors with flat and flared termination points each having an ovoid prominence centrally extending there from. Coupling the truncated ovoid prominence of each conductor with the cylindrically shaped optically active semiconductor causes the semiconductor to cylindrically taper to a triple junction circular line at the base of each prominence where the metal conductor conjoins with the semiconductor and a third medium such as epoxy or air. Tapering the semiconductor at the triple junction inhibits carrier formation and injection at the triple junction and thereby enables greater current carrying capacity through and greater sensitivity of the bulk area of the optically active medium.

Ascites interferes with the activity of lurbinectedin and trabectedin: Potential role of their binding to alpha 1-acid glycoprotein.

PubMed

Erba, E; Romano, M; Gobbi, M; Zucchetti, M; Ferrari, M; Matteo, C; Panini, N; Colmegna, B; Caratti, G; Porcu, L; Fruscio, R; Perlangeli, M V; Mezzanzanica, D; Lorusso, D; Raspagliesi, F; D'Incalci, M

2017-11-15

Trabectedin and its analogue lurbinectedin are effective drugs used in the treatment of ovarian cancer. Since the presence of ascites is a frequent event in advanced ovarian cancer we asked the question whether ascites could modify the activity of these compounds against ovarian cancer cells. The cytotoxicity induced by trabectedin or lurbinectedin against A2780, OVCAR-5 cell lines or primary culture of human ovarian cancer cells was compared by performing treatment in regular medium or in ascites taken from either nude mice or ovarian cancer patients. Ascites completely abolished the activity of lurbinectedin at up to 10nM (in regular medium corresponds to the IC90), strongly reduced that of trabectedin, inhibited the cellular uptake of lurbinectedin and, to a lesser extent, that of trabectedin. Since α1-acid glycoprotein (AGP) is present in ascites at relatively high concentrations, we tested if the binding of the drugs to this protein could be responsible for the reduction of their activity. Adding AGP to the medium at concentration range of those found in ascites, we reproduced the anticytotoxic effect of ascites. Erythromycin partially restored the activity of the drugs, presumably by displacing them from AGP. Equilibrium dialysis experiments showed that both drugs bind AGP, but the affinity of binding of lurbinectedin was much greater than that of trabectedin. KD values are 8±1.7 and 87±14nM for lurbinectedin and trabectedin, respectively. The studies intimate the possibility that AGP present in ascites might reduce the activity of lurbinectedin and to a lesser extent of trabectedin against ovarian cancer cells present in ascites. AGP plasma levels could influence the distribution of these drugs and thus they should be monitored in patients receiving these compounds. Copyright © 2017 Elsevier Inc. All rights reserved.

Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells.

PubMed

Niu, Bowen; Li, Bo; Wu, Chongyang; Wu, Jiang; Yan, Yuan; Shang, Rui; Bai, Chunling; Li, Guangpeng; Hua, Jinlian

2016-11-22

Melatonin has been reported to be an important endogenous hormone for regulating neurogenesis, immunityand the biological clock. Recently, the effects of melatonin on neural stem cells (NSCs), mesenchymal stem cells(MSCs), and induced pluripotent stem cells(iPSCs) have been reported; however, the effects of melatonin on spermatogonia stem cells (SSCs) are not clear. Here, 1μM and 1nM melatonin was added to medium when goat SSCs were cultured in vitro, the results showed that melatonin could increase the formation and size of SSC colonies. Real-time quantitative PCR (QRT-PCR) and western blot analysis showed that the expression levels of SSC proliferation and self-renewal markers were up-regulated. Meanwhile, QRT-PCR results showed that melatonin inhibit the mRNA expression level of SSC differentiation markers. ELISA analysis showed an obvious increase in the concentration of GDNF (a niche factor secreted by Sertoli cells) in the medium when treated with melatonin. Meanwhile, the phosphorylation level of AKT, a downstream of GDNF-GFRa1-RET pathway was activated. In conclusion, melatonin promotes goat SSC proliferation by stimulating GDNF production in Sertoli cells.

Recent Results on SNRs and PWNe from the Fermi Large Area Telescope

NASA Technical Reports Server (NTRS)

Hays, Elizabeth

2010-01-01

a) Symbiotic Binary System: White dwarf + red giant system. b) Nova: White dwarf builds up mass envelope to the point of thermonuclear fusion. c) Dramatic increase in visual magnitude. d) Recurrent Nova? e) Hints but no strong confirmation of previous nova f) Pre-nova activity: 1) White dwarf shows ongoing variability at level of several in magnitude. 2) V407 Cyg companion is a Mira star showing variability at level of several in magnitude. g) Origin of the gamma rays? 1) Strong shock propagating into dense medium around giant star land stellar wind. 2) Pion decay or electron processes?

Active medium gain study of electric-discharge oxygen-iodine laser

NASA Astrophysics Data System (ADS)

Kolobyanin, Yuriy; Adamenkov, Yuriy; Vyskubenko, Boris; Goryachev, Leonid; Ilyin, Sergey; Kalashnik, Anatoliy; Rakhimova, Tatiana; Rogozhnikov, Georgiy

2007-05-01

The paper reports on experimental studies of the active medium gain in supersonic electric-discharge oxygen-iodine laser (DOIL) based on traveling mw discharge. The measurements have included: absolute concentration, yield, and energy efficiency of production of SO in pure oxygen and oxygen-helium mixes at an oxygen partial pressure 3 to 15 Torr. For the gas flow to get rid of atomic oxygen, both heterogeneous mercury oxide coatings of the tube walls and homogeneous additives to the work mix, such as nitrogen oxide, have been used. The active medium of DOIL was formed using a nozzle array of the type of ejector sized as 10*50 mm2. The singlet oxygen-helium mix was supplied through three rows of sonic cylindrical nozzles, while the iodine-carrier gas mix - through two rows of supersonic conical nozzles with a half-opening angle of 10°(arc). The gas-phase iodine was produced in a quartz cell filled with iodine crystals. Room-temperature iodine vapors were picked up with a carrier gas (nitrogen or helium) and thus delivered into the nozzle array. The active medium was investigated by the high-resolution laser diode spectroscopy approach that used the laser type Vortex 6025 purchased from New Focus, Inc. The laser medium gain factor was determined by the intra-cavity approach having a sensitivity about 1*10 -6 cm -1. The static temperature of the medium was determined from the measurements of gain half-width. The gain of the active medium of electric-discharge OIL has been investigated. The DOIL in use was operating on a mix composed as O II:He=1:1 at a total pressure of 6 Torr and flowrate - about 1 mmol/s. With helium as an iodine carrier gas at a flowrate ~3 mmol/s, we have recorded a positive gain in the DOIL medium.

Correlation of endothelin-1 concentration and angiotensin-converting enzyme activity with the staging of liver fibrosis.

PubMed

Kardum, Dusko; Fabijanić, Damir; Lukić, Anita; Romić, Zeljko; Petrovecki, Mladen; Bogdanović, Zoran; Jurić, Klara; Urek-Crncević, Marija; Banić, Marko

2012-06-01

Increased serum angiotensin-converting enzyme (SACE) activity and serum concentration of endothelin-1 (ET-1) were found in liver cirrhosis. We investigated a correlation between the different stages of liver fibrosis and SACE activity and serum ET-1 concentration. Seventy patients with pathohistologically established chronic liver disease were divided in three groups according to Ishak criteria for liver fibrosis: minimal fibrosis (Ishak score 0-1, n =20), medium fibrosis (Ishak score 2-5, n=20) and cirrhosis (Ishak score 6, n=30). SACE activity and ET-1 concentration were determined using commercial ELISA kits. SACE activity and ET-1 concentrations were proportional to the severity of disease, the highest being in patients with liver cirrhosis. Maximal increase in SACE activity was found between minimal and medium fibrosis while maximal increase in ET-1 concentration was revealed between medium fibrosis and cirrhosis. The analysis of the Receiver Operating Characteristic (ROC) curve for SACE activity suggested a cut-off value to separate minimal from medium fibrosis at 59.00 U/L (sensitivity 100%, specificity 64.7%). The cut-off value for serum ET-1 concentration to separate medium fibrosis from cirrhosis was 12.4 pg/mL (sensitivity 96.8%, specificity 94.4%). A positive correlation between SACE activity and ET-1 concentration was registered (Spearman's ñ = 0.438, p = 0.004). Both SACE activity and ET-1 concentration were increased in all stages of liver fibrosis. Cut-off points for SACE activity and ET-1 concentration could be a biochemical marker for the progression of fibrosis. Positive correlation between SACE activity and ET-1 concentration might indicate their interaction in the development of liver cirrhosis.

77 FR 28496 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

Federal Register 2010, 2011, 2012, 2013, 2014

2012-05-15

... daily retention limit would be the default retention limit of one large medium or giant BFT (measuring... default level of one large medium or giant BFT as follows: Two large medium or giant BFT for the January subquota period (75 FR 79309, December 20, 2010); three large medium or giant BFT for June through November...

Somatic embryogenesis and plant regeneration from cell suspension cultures of Cucumis sativus L.

PubMed

Chee, P P; Tricoli, D M

1988-06-01

A procedure for the regeneration of whole cucumber plants (Cucumis sativus L. cv. Poinsett 76) by embryogenesis from cell suspension cultures is described. Embryogenic callus was initiated from the primary leaves of 14-17 day old plants. Suspension cultures of embryogenic cells were grown in liquid Murashige and Skoog basal medium containing 5 uM 2,4,5-trichlorophenoxyacetic acid and 4 uM 6-benzylaminopurine. Suspension cultures were composed of a population of cells that were densely cytoplasmic and potentially embryogenic. Differentiation of embryos was enhanced by washing the suspension culture cells with MS basal medium containing 0.5% activated charcoal and twice with MS basal medium followed by liquid shake cultures in MS basal medium. Sixty to 70 percent of the embryos prewashed with activated charcoal germinated into plantlets with normal morphology. Embryos obtained from suspension cultured cells without prewashing with activated charcoal organized into plantlets with abnormal primary leaves. Morphologically normal plantlets were obtained by excising the shoot tips and transferring them to fresh medium.

Algicidal Effects of Prodigiosin on the Harmful Algae Phaeocystis globosa

PubMed Central

Zhang, Huajun; Peng, Yun; Zhang, Su; Cai, Guanjing; Li, Yi; Yang, Xujun; Yang, Ke; Chen, Zhangran; Zhang, Jun; Wang, Hui; Zheng, Tianling; Zheng, Wei

2016-01-01

Phaeocystis globosa blooms can have negative effects on higher trophic levels in the marine ecosystem and consequently influence human activities. Strain KA22, identified as the bacterium Hahella, was isolated from coastal surface water and used to control P. globosa growth. A methanol extract from the bacterial cells showed strong algicidal activity. After purification, the compound showed a similar structure to prodigiosin when identified with Q-Exactive Orbitrap MS and nuclear magnetic resonance spectra. The compound showed algicidal activity against P. globosa with a 50% Lethal Dose (LD50) of 2.24 μg/mL. The prodigiosin was stable under heat and acid environment, and it could be degraded under alkaline environment and natural light condition. The growth rates of strain KA22 was fast in 2216E medium and the content of prodigiosin in this medium was more than 70 μg/mL after 16 h incubation. The compound showed particularly strong algicidal activity against Prorocentrum donghaiense, P. Globosa, and Heterosigma akashiwo, but having little effect on three other phytoplankton species tested. The results of our research could increase our knowledge on harmful algal bloom control compound and lead to further study on the mechanisms of the lysis effect on harmful algae. PMID:27199932

Comparative study on digestive lipase activities on the self emulsifying excipient Labrasol, medium chain glycerides and PEG esters.

PubMed

Fernandez, Sylvie; Jannin, Vincent; Rodier, Jean-David; Ritter, Nicolas; Mahler, Bruno; Carrière, Frédéric

2007-05-01

Labrasol is a lipid-based self-emulsifying excipient used in the preparation of lipophilic drugs intended for oral delivery. It is mainly composed of PEG esters and glycerides with medium acyl chains, which are potential substrates for digestive lipases. The hydrolysis of Labrasol by porcine pancreatic extracts, human pancreatic juice and several purified digestive lipases was investigated in the present study. Classical human pancreatic lipase (HPL) and porcine pancreatic lipase, which are the main lipases involved in the digestion of dietary triglycerides, showed very low levels of activity on the entire Labrasol excipient as well as on separated fractions of glycerides and PEG esters. On the other hand, gastric lipase, pancreatic lipase-related protein 2 (PLRP2) and carboxyl ester hydrolase (CEH) showed high specific activities on Labrasol. These lipases were found to hydrolyze the main components of Labrasol (PEG esters and monoglycerides) used as individual substrates, whereas these esters were found to be poor substrates for HPL. The lipolytic activity of pancreatic extracts and human pancreatic juice on Labrasol(R) is therefore mainly due to the combined action of CEH and PLRP2. These two pancreatic enzymes, together with gastric lipase, are probably the main enzymes involved in the in vivo lipolysis of Labrasol taken orally.

A Transformerless Hybrid Active Filter Capable of Complying with Harmonic Guidelines for Medium-Voltage Motor Drives

NASA Astrophysics Data System (ADS)

Kondo, Ryota; Akagi, Hirofumi

This paper presents a transformerless hybrid active filter that is integrated into medium-voltage adjustable-speed motor drives for fans, pumps, and compressors without regenerative braking. The authors have designed and constructed a three-phase experimental system rated at 400V and 15kW, which is a downscaled model from a feasible 6.6-kV 1-MW motor drive system. This system consists of the hybrid filter connecting a passive filter tuned to the 7th harmonic filter in series with an active filter that is based on a three-level diode-clamped PWM converter, as well as an adjustable-speed motor drive in which a diode rectifier is used as the front end. The hybrid filter is installed on the ac side of the diode rectifier with no line-frequency transformer. The downscaled system has been exclusively tested so as to confirm the overall compensating performance of the hybrid filter and the filtering performance of a switching-ripple filter for mitigating switching-ripple voltages produced by the active filter. Experimental results verify that the hybrid filter achieves harmonic compensation of the source current in all the operating regions from no-load to the rated-load conditions, and that the switching-ripple filter reduces the switching-ripple voltages as expected.

Glucosinolate biosynthesis in hairy root cultures of broccoli (Brassica oleracea var. italica).

PubMed

Kim, Sun-Ju; Park, Woo Tae; Uddin, Md Romij; Kim, Yeon Bok; Nam, Sang-Yong; Jho, Kwang Hyun; Park, Sang Un

2013-02-01

Here we present previously unreported glucosinolate production by hairy root cultures of broccoli (B. oleracea var. italica). Growth media greatly influenced the growth and glucosinolate content of hairy root cultures of broccoli. Seven glucosinolates, glucoraphanin, gluconapin, glucoerucin, glucobrassicin, 4-methoxyglucobrassicin, gluconasturtiin, and neoglucobrassicin, were identified by analysis of the broccoli hairy root cultures. Both half and full strength B5 and SH media enabled the highest accumulation of glucosinolates. In most cases, the levels of glucosinolates were higher in SH and BS media. Among the 7 glucosinolates, the accumulation of neoglucobrassicin was very high, irrespective of growth medium. The neoglucobrassicin content was 7.4-fold higher in SH medium than 1/2 MS, in which its level was the lowest. The 1/2 B5 medium supported the production of the highest amounts of glucobrassicin and 4-methoxyglucobrassicin, the levels for which were 36.2- and 7.9- fold higher, respectively, than their lowest content in 1/2 MS medium. The 1/2 SH medium enabled the highest accumulation of glucoraphanin and gluconapin in the broccoli hairy root cultures, whose levels were 1.8- and 4.6-fold higher, respectively, than their lowest content in 1/2 MS medium. Our results suggest that hairy root cultures of broccoli could be a valuable alternative approach for the production of glucosinolate compounds.

ENZYME ACTIVITIES DURING THE ASEXUAL CYCLE OF NEUROSPORA CRASSA

PubMed Central

Stine, G. J.

1968-01-01

Three enzymes, (a) nicotinamide adenine diphosphate-dependent glutamic dehydrogenase (NAD enzyme), (b) nictoinamide adenine triphosphate-dependent glutamic dehydrogenase (NADP enzyme), and (c) nicotinamide-adenine dinucleotidase (NADase), were measured in separate extracts of Neurospora crassa grown in Vogel's medium N and medium N + glutamate. Specific activities and total units per culture of each enzyme were determined at nine separate intervals phased throughout the asexual cycle. The separate dehydrogenases were lowest in the conidia, increased slowly during germination, and increased rapidly during logarithmic mycelial growth. The amounts of these enzymes present during germination were small when compared with those found later during the production of the conidiophores. The NAD enzyme may be necessary for pregermination synthesis. The NADP-enzyme synthesis was associated with the appearance of the germ tube. Although higher levels of the dehydrogenases in the conidiophores resulted in more enzyme being found in the differentiated conidia, the rate of germination was uneffected. The greatest activity for the NADase enzyme was associated with the conidia, early phases of germination, and later production of new conidia. NADase decreased significantly with the onset of logarithmic growth, remained low during the differentiation of conidiophores, and increased considerably as the conidiophores aged. PMID:4384627

Biohybrid Polymer-Antimicrobial Peptide Medium against Enterococcus faecalis

PubMed Central

Eckhard, Lea H.; Sol, Asaf; Abtew, Ester; Shai, Yechiel; Domb, Abraham J.

2014-01-01

Antimicrobial peptides (AMPs) are conserved evolutionary components of the innate immune system that are being tested as alternatives to antibiotics. Slow release of AMPs using biodegradable polymers can be advantageous in maintaining high peptide levels for topical treatment, especially in the oral environment in which dosage retention is challenged by drug dilution with saliva flow and by drug inactivation by salivary enzymatic activity. Enterococcus faecalis is a multidrug resistant nosocomial pathogen and a persistent pathogen in root canal infections. In this study, four ultra-short lipopeptides (C16-KGGK, C16-KLLK, C16-KAAK and C16-KKK) and an amphipathic α-helical antimicrobial peptide (Amp-1D) were tested against E. faecalis. The antibacterial effect was determined against planktonic bacteria and bacteria grown in biofilm. Of the five tested AMPs, C16-KGGK was the most effective. Next C16-KGGK was formulated with one of two polymers poly (lactic acid co castor oil) (DLLA) or ricinoleic acid-based poly (ester-anhydride) P(SA-RA). Peptide-synthetic polymer conjugates, also referred to as biohybrid mediums were tested for antibacterial activity against E. faecalis grown in suspension and in biofilms. The new formulations exhibited strong and improved anti- E. faecalis activity. PMID:25279943

[Influence of tap water treatment on perfluorinated compounds residue in the dissolved phase].

PubMed

Zhang, Hong; Chen, Qing-wu; Wang, Xin-xuan; Chai, Zhi-fang; Shen, Jin-can; Yang, Bo; Liu, Guo-qing

2013-09-01

To study the perfluorinated compounds (PFCs) residues through water treatments including flocculation, sedimentation, sand filtration, ozonation with activated carbon and chlorination, as well as the seasonal variation of PFCs in the raw water of waterworks, 13 PFCs species in the dissolved phase of raw water, finished water, as well as the water samples after flocculation, sedimentation, sand filtration, and ozonation with activated carbon filtration were measured by the high performance liquid chromatography-tandem mass spectrometry combined with solid phase extraction. Results indicated that sigma PFCs residue in water was higher in spring and summer than that in fall and winter. The vast majority of PFCs in samples were of short and medium chains (C < or = 10), and perfluorooctane sulfonate was the most typical residue species. Among the five water treatment stages, sedimentation, sand filtration and ozonation with activated carbon filtration can remove PFCs, while flocculation and chlorination significantly raise the levels of short- (C < or = 6) and medium-chain (10 > or = C > or = 7) PFCs, respectively, causing sigma PFCs increase in finished water by 10%-44% compared to raw water. However, the PFCs residues in finished water are still far below their limit values, posing no threat against human health.

Chromospherically Active Stars in the RAVE Survey

NASA Astrophysics Data System (ADS)

Žerjal, M.; Zwitter, T.; Matijevič, G.; Strassmeier, K. G.

2014-01-01

We present a qualitative characterization of activity levels of a large database of ~44,000 candidate RAVE stars (unbiased, magnitude limited medium resolution survey) that show chromospheric emission in the Ca II infrared triplet and this vastly enlarges previously known samples. Our main motivation to study these stars is the anti-correlation of chromospheric activity and stellar ages that could be calibrated using stellar clusters with known ages. Locally linear embedding used for a morphological classification of spectra revealed 53,347 cases with a suggested emission component in the calcium lines. We analyzed a subsample of ~44,000 stars with S/N>20 using a spectral subtraction technique where observed reference spectra of inactive stars were used as templates instead of synthetic ones. Both the equivalent width of the excess emission for each calcium line and their sum is derived for all candidate active stars with no respect to the origin of their emission flux. ~17,800 spectra show a detectable chromospheric flux with at least 2 σ confidence level. The overall distribution of activity levels shows a bimodal shape, with the first peak coinciding with inactive stars and the second with the pre-main-sequence cases.

High calcium to phosphorus ratio impairs growth and bone mineralization in Pekin ducklings.

PubMed

Zhu, Y W; Wen, J; Jiang, X X; Wang, W C; Yang, L

2018-04-01

Two experiments were conducted to investigate the effect of high dietary calcium (Ca) level on growth performance, Ca and phosphorus (P) metabolism, and nutrient utilization in ducklings subjected to normal and low P levels in diets. A completely randomized design was used with a factorial arrangement of 2 total dietary P levels [normal-P (0.60%) and low-P (0.45%) groups] × 4 dietary Ca levels [low-Ca (0.55%), normal-Ca (0.75%), medium-Ca (0.95%) and high-Ca (1.15%) groups)]. Compared to normal-P group, low-P group had lower (P < 0.05) final body weight (BW), average daily gain (ADG), and average daily feed intake (ADFI) and reduced (P < 0.05) serum Ca and P levels, bone Ca, P, and ash content, and bone mineral density in ducklings during the starter period. Under the low-P group, birds from high-Ca group had lower (P < 0.05) final BW, ADG, ADFI, bone ash content, bone mineral density, and the utilization of energy, Ca, and P than those from low-Ca, normal-Ca, and medium-Ca groups. Our results indicate that high-Ca diet induced greater growth suppression and bone mineralization loss in ducklings fed a low-P diet. The aggravated negative effect of high dietary Ca level with a low P level might be related to the elevated serum alkaline phosphatase activity and the reduced utilization of energy, Ca, and P.

Cellular Antioxidant and Anti-Inflammatory Effects of Coffee Extracts with Different Roasting Levels.

PubMed

Jung, Soohan; Kim, Min Hyung; Park, Jae Hee; Jeong, Yoonhwa; Ko, Kwang Suk

2017-06-01

During roasting, major changes occur in the composition and physiological effects of coffee beans. In this study, in vitro antioxidant effects and anti-inflammatory effects of Coffea arabica green coffee extracts were investigated at different roasting levels corresponding to Light, Medium, City, and French roast. Total caffeine did not show huge difference according to roasting level, but total chlorogenic acid contents were higher in light roasted coffee extract than other roasted groups. In addition, light roasted coffee extract had the highest antioxidant activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. To determine the in vitro antioxidant property, coffee extracts were used to treat AML-12 cells. Intracellular glutathione (GSH) concentration and mRNA expression levels of genes related to GSH synthesis were negatively related to roasting levels. The anti-inflammatory effects of coffee extracts were investigated in lipopolysaccharide-treated RAW 264.7 macrophage cells. The cellular antioxidant activity of coffee extracts exhibited similar patterns as the AML-12 cells. The expression of mRNA for tumor necrosis factor-alpha and interleukin-6 was decreased in cells treated with the coffee extracts and the expression decreased with increasing roasting levels. These data suggest that coffee has physiological antioxidant and anti-inflammatory activities and these effects are negatively correlated with roasting levels in the cell models.

Proof-of-Concept of a Revolutionary Cooling Shirt with a Thermal Manikin

DTIC Science & Technology

2013-09-01

involving high levels of physical activity or exposure to high temperature environments including military, fire protection, and sports . Battlefield...spandex. Pockets of the same material were created to fully encapsulate the lightweight cryotherapy pad. The cooling medium is flame resistant to 815°C...effect on the manikin. Per manufacturer recommendation, the cryotherapy pads were stored in a refrigerator at 10°C to obtain a maximum effect. New pads

An overview of the measurements of soil moisture and modeling of moisture flux in FIFE

NASA Technical Reports Server (NTRS)

Wang, J. R.

1992-01-01

Measurements of soil moisture and calculations of moisture transfer in the soil medium and at the air-soil interface were performed over a 15-km by 15-km test site during FIFE in 1987 and 1989. The measurements included intensive soil moisture sampling at the ground level and surveys at aircraft altitudes by several passive and active microwave sensors as well as a gamma radiation device.

The effects of a physical activity intervention on employees in small and medium enterprises: a mixed methods study.

PubMed

Edmunds, Sarah; Stephenson, Duncan; Clow, Angela

2013-01-01

Workplaces have potential as a setting for physical activity promotion but evidence of the effectiveness of intervention programmes in small and medium sized enterprises is limited. This paper reports the impact of an intervention which trained existing employees to promote physical activity to their colleagues. Eighty-nine previously low-active employees from 17 small and medium sized organisations participated. A mixed methods evaluation design was used. Quantitative data were collected at baseline and 6 months later using an online questionnaire. Qualitative data from a series of 6 focus groups were analysed. Repeated measures t-tests showed significant increases over time in physical activity, general health rating, satisfaction with life and positive mood states. There were significant decreases in body mass index (BMI), perceived stress, negative mood states and presenteeism. There was no change in absenteeism. Analysis of focus group data provided further insight into the impact of the intervention. Five major themes emerged: awareness of physical activity; sustaining physical activity behaviour change; improved health and well-being; enhanced social networks; and embedding physical activity in the workplace culture. This study shows it is feasible and effective to train employees in small and medium sized enterprises to support their colleagues in physical activity behaviour change.

Cultivation characteristics and gene expression profiles of Aspergillus oryzae by membrane-surface liquid culture, shaking-flask culture, and agar-plate culture.

PubMed

Imanaka, Hiroyuki; Tanaka, Soukichi; Feng, Bin; Imamura, Koreyoshi; Nakanishi, Kazuhiro

2010-03-01

We cultivated a filamentous fungus, Aspergillus oryzae IAM 2706 by three different cultivation methods, i.e., shaking-flask culture (SFC), agar-plate culture (APC), and membrane-surface liquid culture (MSLC), to elucidate the differences of its behaviors by different cultivation methods under the same media, by measuring the growth, secretion of proteases and alpha-amylase, secreted protein level, and gene transcriptional profile by the DNA microarray analysis. The protease activities detected by MSLC and APC were much higher than that by SFC, using both modified Czapek-Dox (mCD) and dextrin-peptone-yeast extract (DPY) media. The alpha-amylase activity was detected in MSLC and APC in a much larger extent than that in SFC when DPY medium was used. On the basis of SDS-PAGE analyses and N-terminal amino acid sequences, 6 proteins were identified in the supernatants of the culture broths using DPY medium, among which oryzin (alkaline protease) and alpha-amylase were detected at a much higher extent for APC and MSLC than those for SFC while only oryzin was detected in mCD medium, in accordance with the activity measurements. A microarray analysis for the fungi cultivated by SFC, APC, and MSLC using mCD medium was carried out to elucidate the differences in the gene transcriptional profile by the cultivation methods. The gene transcriptional profile obtained for the MSLC sample showed a similar tendency to the APC sample while it was quite different from that for the SFC sample. Most of the genes specifically transcribed in the MSLC sample versus those in the SFC sample with a 10-fold up-regulation or higher were unknown or predicted proteins. However, transcription of oryzin gene was only slightly up-regulated in the MSLC sample and that of alpha-amylase gene, slightly down-regulated. Copyright 2009 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Nutrients versus emerging contaminants-Or a dynamic match between subsidy and stress effects on stream biofilms.

PubMed

Aristi, I; Casellas, M; Elosegi, A; Insa, S; Petrovic, M; Sabater, S; Acuña, V

2016-05-01

Freshwater ecosystems are threatened by multiple anthropogenic stressors, which might be differentiated into two types: those that reduce biological activity at all concentrations (toxic contaminants), and those that subsidize biological activity at low concentrations and reduce it at high concentrations (assimilable contaminants). When occurring in mixtures, these contaminants can have either antagonistic, neutral or synergistic effects; but little is known on their joint effects. We assessed the interaction effects of a mixture of assimilable and toxic contaminants on stream biofilms in a manipulative experiment using artificial streams, and following a factorial design with three nutrient levels (low, medium or high) and either presence or absence of a mixture of emerging contaminants (ciprofloxacin, erythromycin, diclofenac, methylparaben, and sulfamethoxazole). We measured biofilm biomass, basal fluorescence, gross primary production and community respiration. Our initial hypotheses were that biofilm biomass and activity would: increase with medium nutrient concentrations (subsidy effect), but decrease with high nutrient concentrations (stress effect) (i); decrease with emerging contaminants, with the minimum decrease at medium nutrient concentrations (antagonistic interaction between nutrients subsidy and stress by emerging contaminants) and the maximum decrease at high nutrient concentrations (synergistic interaction between nutrients and emerging contaminants stress) (ii). All the measured variables responded linearly to the available nutrients, with no toxic effect at high nutrient concentrations. Emerging contaminants only caused weak toxic effects in some of the measured variables, and only after 3-4 weeks of exposure. Therefore, only antagonistic interactions were observed between nutrients and emerging contaminants, as medium and high nutrient concentrations partly compensated the harmful effects of emerging contaminants during the first weeks of the experiment. Our results show that contaminants with a subsidy effect can alleviate the effects of toxic contaminants, and that long-term experiments are required to detect stress effects of emerging contaminants at environmentally relevant concentrations. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Plastic scintillators in coincidence for the study of multi-particle production of sea level cosmic rays in dense medium

NASA Technical Reports Server (NTRS)

Chuang, L. S.; Chan, K. W.; Wada, M.

1985-01-01

Cosmic ray particles at sea level penetrate a thick layer of dense medium without appreciable interaction. These penetrating particles are identified with muons. The only appreciable interaction of muons are by knock on processes. A muon may have single, double or any number of knock on with atoms of the material so that one, two, three or more particles will come out from the medium in which the knock on processes occur. The probability of multiparticle production is expected to decrease with the increase of multiplicity. Measurements of the single, double, and triple particles generated in a dense medium (Fe and Al) by sea level cosmic rays at 22.42 N. Lat. and 114.20 E. Long. (Hong Kong) are presented using a detector composed of two plastic scintillators connected in coincidence.

Place of residence as a factor differentiating physical activity in the life style of Ukrainian students.

PubMed

Bergier, Józef; Bergier, Barbara; Tsos, Anatolii

2016-12-23

Determining the state of physical activity of societies as an important component of a health promoting life style is a very up-to-date problem. Studies of physical activity among students, the future elites in their environments, become of increasing importance. An important problem is the recognition of factors differentiating this activity on the example of place of residence. For this purpose, the study covered 2,125 students (60.8% females and 39.2% males) from the National Institute in Lutsk, Ukraine, aged 17-22 (mean age: 20.4). The method of a diagnostic survey was applied which included the International Physical Activity Questionnaire (IPAQ). The following measures of physical activity according to the place of residence (rural area, small town with a population up to 100,000; medium-size town - 100,000-200,000 inhabitants; large city - over 200,000) were taken into consideration: level of physical activity, self-reported physical fitness, sports disciplines practiced by the respondents, and those which they would like to practice, and the BMI, and leisure time possessed. The study showed that the place of residence positively differentiated physical activity among students from medium-size towns and rural areas, compared to their contemporaries from small towns and large cities. Significant differences were also found with respect to the BMI, which was significantly less favourable among respondents from the rural environment. However, no differences were observed between the place of residence for leisure time, self-reported physical activity, and forms of physical activity practiced, and those which the respondents would like to practice.

Metabolic acidosis increases fibroblast growth factor 23 in neonatal mouse bone

PubMed Central

Culbertson, Christopher D.; Kyker-Snowman, Kelly; Bushinsky, David A.

2012-01-01

Fibroblast growth factor 23 (FGF23) significantly increases with declining renal function, leading to reduced renal tubular phosphate reabsorption, decreased 1,25-dihydroxyvitamin D, and increased left ventricular hypertrophy. Elevated FGF23 is associated with increased mortality. FGF23 is synthesized in osteoblasts and osteocytes; however, the mechanisms by which it is regulated are not clear. Patients with chronic kidney disease have decreased renal acid excretion leading to metabolic acidosis, which has a direct effect on bone cell activity. We hypothesized that metabolic acidosis would directly increase bone cell FGF23 production. Using cultured neonatal mouse calvariae, we found that metabolic acidosis increased medium FGF23 protein levels as well as FGF23 RNA expression at 24 h and 48 h compared with incubation in neutral pH medium. To exclude that the increased FGF23 was secondary to metabolic acidosis-induced release of bone mineral phosphate, we cultured primary calvarial osteoblasts. In these cells, metabolic acidosis increased FGF23 RNA expression at 6 h compared with incubation in neutral pH medium. Thus metabolic acidosis directly increases FGF23 mRNA and protein in mouse bone. If these results are confirmed in humans with chronic kidney disease, therapeutic interventions to mitigate acidosis, such as bicarbonate administration, may also lower levels of FGF23, decrease left ventricular hypertrophy, and perhaps even decrease mortality. PMID:22647635

Changes in soluble sugar, starch, and alcohol dehydrogenase in Arabidopsis thaliana exposed to N2 diluted atmospheres

NASA Technical Reports Server (NTRS)

Porterfield, D. M.; Crispi, M. L.; Musgrave, M. E.

1997-01-01

Proper exchange of atmospheric gases is important for normal root and shoot metabolism in plants. This study was conducted to determine how restricted air supply affects foliar carbohydrates, while using the marker enzyme alcohol dehydrogenase (ADH) to report on the oxygenation status of the rootzone. Fourteen-day-old Arabidopsis thaliana (L.) Heynh. plants grown singly in 7-ml tubes containing agarified nutrient medium were placed in coupled Magenta vessels and exposed for six days to either ambient air or one of six different air/nitrogen dilutions. Redox potential of the agar medium was measured immediately after harvesting and freezing leaf tissue, and then root systems were quickly extracted from the agar and frozen for subsequent analyses. Redox potential measurements indicated that this series of gas mixtures produced a transition from hypoxia to anoxia in the root zones. Root ADH activity increased at higher rates as the redox potential neared anoxic levels. In contrast, ADH mRNA expression quickly neared its maximum as the medium became hypoxic and showed little further increase as it became anoxic. Foliar carbohydrate levels increased 1.5- to 2-fold with decreased availability of metabolic gases, with starch increasing at higher concentrations of air than soluble carbohydrate. The results serve as a model for plant performance under microgravity conditions, where absence of convective air movement prevents replenishment of metabolic gases.

Serum-Free Medium and Mesenchymal Stromal Cells Enhance Functionality and Stabilize Integrity of Rat Hepatocyte Spheroids

PubMed Central

Bao, Ji; Fisher, James E.; Lillegard, Joseph B.; Wang, William; Amiot, Bruce; Yu, Yue; Dietz, Allan B.; Nahmias, Yaakov; Nyberg, Scott L.

2013-01-01

Long-term culture of hepatocyte spheroids with high ammonia clearance is valuable for therapeutic applications, especially the bioartificial liver. However, the optimal conditions are not well studied. We hypothesized that liver urea cycle enzymes can be induced by high protein diet and maintain on a higher expression level in rat hepatocyte spheroids by serum-free medium (SFM) culture and coculture with mesenchymal stromal cells (MSCs). Rats were feed normal protein diet (NPD) or high protein diet (HPD) for 7 days before liver digestion and isolation of hepatocytes. Hepatocyte spheroids were formed and maintained in a rocked suspension culture with or without MSCs in SFM or 10% serum-containing medium (SCM). Spheroid viability, kinetics of spheroid formation, hepatic functions, gene expression, and biochemical activities of rat hepatocyte spheroids were tested over 14 days of culture. We observed that urea cycle enzymes of hepatocyte spheroids can be induced by high protein diet. SFM and MSCs enhanced ammonia clearance and ureagenesis and stabilized integrity of hepatocyte spheroids compared to control conditions over 14 days. Hepatocytes from high protein diet-fed rats formed spheroids and maintained a high level of ammonia detoxification for over 14 days in a novel SFM. Hepatic functionality and spheroid integrity were further stabilized by coculture of hepatocytes with MSCs in the spheroid microenvironment. These findings have direct application to development of the spheroid reservoir bioartificial liver. PMID:23006214

[The phylogenetic theory of pathology. The arterial hypertension--a test of metabolic disorder. The biological basis of damage of target organs (a lecture)].

PubMed

Titov, V N

2013-05-01

The increase of blood tension is a diagnostic test of disorders of homeostasis, trophology, endoecology and adaptation in paracrine regulated coenosis of cells. This conditions results in disorder of microcirculation in the distal section of arterial race and in compensatory increase of blood tension in its proximal section. The increase of blood tension disturbs the function of paracrine coenosis of cells which have one's own system of hemo- and hydrodynamics such as brain with system of spinal liquor and kidneys with local pool of primary urine. They counteract the rise of blood tension and activate local, humoral system of renin-angiotensin-II increasing peripheral resistance to blood flow. At that, the compensatory blood tension becomes even higher. The aldosterone and natriuretic peptides are functional synergists. So, they preserve and excrete ions of Na+ and support the stability of unified pool of intercellular medium ("Inner Ocean" of organism) where all cells live. The parameters of this pool are limited most strictly in vivo. If at the level of nephron the conditions are formed that can alter the parameters of unified pool of intercellular medium the vasomotor center rises blood tension from the level of organism "forcing" nephrons to re-establish the parameters of this pool and normalize the biological functions and biological reactions. The blood pressure increase under pathology of kidneys is caused because of pathological compensation at the level of organism mediated by vegetal nervous system and dictated by necessity to preserve the parameters of inner medium of organism.

Hydrological and sedimentary controls over fluvial thermal erosion, the Lena River, central Yakutia

NASA Astrophysics Data System (ADS)

Tananaev, Nikita I.

2016-01-01

Water regime and sedimentary features of the middle Lena River reach near Yakutsk, central Yakutia, were studied to assess their control over fluvial thermal erosion. The Lena River floodplain in the studied reach has complex structure and embodies multiple levels varying in height and origin. Two key sites, corresponding to high and medium floodplain levels, were surveyed in 2008 to describe major sedimentary units and properties of bank material. Three units are present in both profiles, corresponding to topsoil, overbank (cohesive), and channel fill (noncohesive) deposits. Thermoerosional activity is mostly confined to a basal layer of frozen channel fill deposits and in general occurs within a certain water level interval. Magnitude-frequency analysis of water level data from Tabaga gauging station shows that a single interval can be deemed responsible for the initiation of thermal action and development of thermoerosional notches. This interval corresponds to the discharges between 21,000 and 31,000 m3 s- 1, observed normally during spring meltwater peak and summer floods. Competence of fluvial thermal erosion depends on the height of floodplain level being eroded, as it acts preferentially in high floodplain banks. In medium floodplain banks, thermal erosion during spring flood is constrained by insufficient bank height, and erosion is essentially mechanical during summer flood season. Bank retreat rate is argued to be positively linked with bank height under periglacial conditions.

[Relationship between the frequency of work-related stress and prevalence of functional dyspepsia in Lima Geriatric Army Hospital].

PubMed

Valenzuela Narváez, Daniel Raúl; Gayoso Cervantes, Milagros

2017-01-01

To determine the relationship between the frequency of work-related stress and prevalence of functional dyspepsia in a sample of 218 military older 50 years in 2010 in Lima Military Hospital Geriatric. Descriptive and explanatory study and for the data collection on stress, were used the Vital Events Scale Holmes-Rahe and clinical records for clinical and upper endoscopy registration that comply the criteria of Rome III for functional dyspepsia. For processing and data analysis the statistical software package SPSS (Statistical Package for Social Sciences) was used. 100% of military showed some level of work stress during the study year; thus, 36.7% had a high level, 31.2% medium or moderate level, and 32.1% had low stress level; these percentages medium and high stress levels accounted for 67.9%. These results establish that job stress is a common discomfort in the study population (tabulated Chi2 = 3.841, chi2 observed = 27,908). Regarding functional dyspepsia prevalence of 37.2%, which indicates that it is a common condition in those military (tabular Z = 1.96, Z c = 9.163) it was determined. There is a significant relationship between the frequency of work-related stress and prevalence of functional dyspepsia in military activity in older than 50 years (tabulated Chi2 = 5.991, chi2 observed = 28,878, contingency coefficient = 0.342).

Systematic evaluation of sericin protein as a substitute for fetal bovine serum in cell culture.

PubMed

Liu, Liyuan; Wang, Jinhuan; Duan, Shengchang; Chen, Lei; Xiang, Hui; Dong, Yang; Wang, Wen

2016-08-17

Fetal bovine serum (FBS) shows obvious deficiencies in cell culture, such as low batch to batch consistency, adventitious biological contaminant risk, and high cost, which severely limit the development of the cell culture industry. Sericin protein derived from the silkworm cocoon has become increasingly popular due to its diverse and beneficial cell culture characteristics. However, systematic evaluation of sericin as a substitute for FBS in cell culture medium remains limited. In this study, we conducted cellular morphological, physiological, and transcriptomic evaluation on three widely used mammalian cells. Compared with cells cultured in the control, those cultured in sericin-substitute medium showed similar cellular morphology, similar or higher cellular overall survival, lower population doubling time (PDT), and a higher percentage of S-phase with similar G2/G1 ratio, indicating comparable or better cell growth and proliferation. At the transcriptomic level, differentially expressed genes between cells in the two media were mainly enriched in function and biological processes related to cell growth and proliferation, reflecting that genes were activated to facilitate cell growth and proliferation. The results of this study suggest that cells cultured in sericin-substituted medium perform as well as, or even better than, those cultured in FBS-containing medium.

Systematic evaluation of sericin protein as a substitute for fetal bovine serum in cell culture

PubMed Central

Liu, Liyuan; Wang, Jinhuan; Duan, Shengchang; Chen, Lei; Xiang, Hui; Dong, Yang; Wang, Wen

2016-01-01

Fetal bovine serum (FBS) shows obvious deficiencies in cell culture, such as low batch to batch consistency, adventitious biological contaminant risk, and high cost, which severely limit the development of the cell culture industry. Sericin protein derived from the silkworm cocoon has become increasingly popular due to its diverse and beneficial cell culture characteristics. However, systematic evaluation of sericin as a substitute for FBS in cell culture medium remains limited. In this study, we conducted cellular morphological, physiological, and transcriptomic evaluation on three widely used mammalian cells. Compared with cells cultured in the control, those cultured in sericin-substitute medium showed similar cellular morphology, similar or higher cellular overall survival, lower population doubling time (PDT), and a higher percentage of S-phase with similar G2/G1 ratio, indicating comparable or better cell growth and proliferation. At the transcriptomic level, differentially expressed genes between cells in the two media were mainly enriched in function and biological processes related to cell growth and proliferation, reflecting that genes were activated to facilitate cell growth and proliferation. The results of this study suggest that cells cultured in sericin-substituted medium perform as well as, or even better than, those cultured in FBS-containing medium. PMID:27531556

Growth and consumption of L-malic acid in wine-like medium by acclimated and non-acclimated cultures of Patagonian Oenococcus oeni strains.

PubMed

Bravo-Ferrada, Bárbara Mercedes; Hollmann, Axel; Brizuela, Natalia; La Hens, Danay Valdés; Tymczyszyn, Elizabeth; Semorile, Liliana

2016-09-01

Five Oenococcus oeni strains, selected from spontaneous malolactic fermentation (MLF) of Patagonic Pinot noir wine, were assessed for their use as MLF starter cultures. After the individual evaluation of tolerance to some stress conditions, usually found in wine (pH, ethanol, SO2, and lysozyme), the behavior of the strains was analyzed in MLO broth with 14 % ethanol and pH 3.5 in order to test for the synergistic effect of high ethanol level and low pH and, finally, in a wine-like medium. Although the five strains were able to grow in MLO broth under low pH and/or high ethanol, they must be acclimated to grow in a wine-like medium. Additionally, glycosidase and tannase activities were evaluated, showing differences among the strains. The potential of the strains to ferment citrate was tested and two of the five strains showed the ability to metabolize this substrate. We did not detect the presence of genes encoding histidine, tyrosine descarboxylase, and putrescine carbamoyltransferase. All the strains tested exhibited good growth capacity and ability to consume L-malic acid in a wine-like medium after cell acclimation, and each of them showed a particular enzyme profile, which might confer different organoleptic properties to the wine.

Pseudomonas aeruginosa mutants resistant to urea inhibition of growth on acetanilide.

PubMed Central

Gregoriou, M; Brown, P R; Tata, R

1977-01-01

Pseudomonas aeruginosa AI 3 was able to grow in medium containing acetanilide (N-phenylacetamide) as a carbon source when NH4+ was the nitrogen source but not when urea was the nitrogen source. AIU mutants isolated from strain AI 3 grew on either medium. Urease levels in bacteria grown in the presence of urea were 10-fold lower when NH4+ or acetanilide was also in the medium, but there were no apparent differences in urease or its synthesis between strain AI 3 and mutant AIU 1N. The first metabolic step in the acetanilide utlization is catalyzed by an amidase. Amidases in several AIU strains showed altered physiochemical properties. Urea inhibited amidase in a time-dependent reaction, but the rates of the inhibitory reaction with amidases from the AIU mutants were slower than with AI 3 amidase. The purified amidase from AIU 1N showed a marked difference in its pH/activity profile from that obtained with purified AI 3 amidase. These observations indicate that the ability of strain AIU 1N and the other mutants to grow on acetanilide/urea medium is associated with a mutation in the amidase structural gene; this was confirmed for strain AIU 1N by transduction. PMID:410788

DOE Office of Scientific and Technical Information (OSTI.GOV)

Spier, C.E.; Little, D.E.; Trim, S.C.

We investigated activity patterns of 17 elementary school students aged 10-12, and 19 high school students aged 13-17, in suburban Los Angeles during the oxidant pollution season. Individuals' relationships between ventilation rate (VR) and heart rate (HR) were calibrated' in supervised outdoor walking/jogging. Log VR was consistently proportional to HR; although calibrations' were limited by a restricted range of exercise, and possibly by artifact due to mouthpiece breathing, which may cause overestimation of VR at rest. Each subject then recorded activities in diaries, and recorded HR once per minute by wearing Heart Watches, over 3 days (Saturday-Monday). For each activitymore » the subject estimated a breathing rate--slow (slow walking), medium (fast walking), or fast (running). VR ranges for each breathing rate and activity type were estimated from HR recordings. High-school students' diaries showed their aggregate distribution of waking hours as 68% slow inside, 8% slow outside, 10% medium inside, 9% medium outside, 1.5% fast inside, 1.5% fast outside. Elementary students' distribution was 47% slow inside, 15% slow outside, 20% medium inside, 12% medium outside, 2.5% fast inside, 3.5% fast outside. Sleep occupied 38% of high-school students' and 40% of elementary students' time; HR were generally lower in sleep than in slow waking activity. High school students' mean VR estimates were 13 L/min for slow breathing, 18 for medium, and 23 for fast; elementary students' were 14 slow, 18 medium, and 19 fast. VR distributions were approximately lognormal. Maximum estimated VR were approximately 70 L/min in elementary and approximately 100 L/min in high school students. Compared to adults studied similarly, students reported more medium or fast breathing, and had equal or higher VR estimates during slow and medium breathing despite their smaller size. These results suggest that, relative to body size, young people inhale larger doses of outdoor air pollutants than adults.« less

Decreased nitric oxide levels stimulate osteoclastogenesis and bone resorption both in vitro and in vivo on the chick chorioallantoic membrane in association with neoangiogenesis.

PubMed

Collin-Osdoby, P; Rothe, L; Bekker, S; Anderson, F; Osdoby, P

2000-03-01

High nitric oxide (NO) levels inhibit osteoclast (OC)-mediated bone resorption in vivo and in vitro, and nitrate donors protect against estrogen-deficient bone loss in postmenopausal women. Conversely, decreased NO production potentiates OC bone resorption in vitro and is associated with in vivo bone loss in rats and humans. Previously, we reported that bone sections from rats administered aminoguanidine (AG), a selective inhibitor of NO production via inducible NO synthase, exhibited both increased OC resorptive activity as well as greater numbers of OC. Here, we investigated further whether AG promoted osteoclastogenesis, in addition to stimulating mature OC function, using a modified in vivo chick chorioallantoic membrane (CAM) system and an in vitro chick bone marrow OC-like cell developmental model. AG, focally administered in small agarose plugs placed directly adjacent to a bone chip implanted on the CAM, dose-dependently elicited neoangiogenesis while stimulating the number, size, and bone pit resorptive activity of individual OC ectopically formed in vivo. In addition to enhancing OC precursor recruitment via neoangiogenesis, AG also exerted other vascular-independent effects on osteoclastogenesis. Thus, AG promoted the in vitro fusion and formation from bone marrow precursor cells of larger OC-like cells that contained more nuclei per cell and exhibited multiple OC differentiation markers. AG stimulated development was inversely correlated with declining medium nitrite levels. In contrast, three different NO donors each dose-dependently inhibited in vitro OC-like cell development while raising medium nitrite levels. Therefore, NO sensitively regulates OC-mediated bone resorption through affecting OC recruitment (angiogenesis), formation (fusion and differentiation), and bone resorptive activity in vitro and in vivo. Possibly, the stimulation of neoangiogenesis and OC-mediated bone remodeling via AG or other pro-angiogenic agents may find clinical applications in reconstructive surgery, fracture repair, or the treatment of avascular necrosis.

PreImplantation Factor (PIF) correlates with early mammalian embryo development-bovine and murine models

PubMed Central

2011-01-01

Background PreImplantation Factor (PIF), a novel peptide secreted by viable embryos is essential for pregnancy: PIF modulates local immunity, promotes decidual pro-adhesion molecules and enhances trophoblast invasion. To determine the role of PIF in post-fertilization embryo development, we measured the peptide's concentration in the culture medium and tested endogenous PIF's potential trophic effects and direct interaction with the embryo. Methods Determine PIF levels in culture medium of multiple mouse and single bovine embryos cultured up to the blastocyst stage using PIF-ELISA. Examine the inhibitory effects of anti-PIF-monoclonal antibody (mAb) added to medium on cultured mouse embryos development. Test FITC-PIF uptake by cultured bovine blastocysts using fluorescent microscopy. Results PIF levels in mouse embryo culture medium significantly increased from the morula to the blastocyst stage (ANOVA, P = 0.01). In contrast, atretic embryos medium was similar to the medium only control. Detectable - though low - PIF levels were secreted already by 2-cell stage mouse embryos. In single bovine IVF-derived embryos, PIF levels in medium at day 3 of culture were higher than non-cleaving embryos (control) (P = 0.01) and at day 7 were higher than day 3 (P = 0.03). In non-cleaving embryos culture medium was similar to medium alone (control). Anti-PIF-mAb added to mouse embryo cultures lowered blastocyst formation rate 3-fold in a dose-dependent manner (2-way contingency table, multiple groups, X2; P = 0.01) as compared with non-specific mouse mAb, and medium alone, control. FITC-PIF was taken-up by cultured bovine blastocysts, but not by scrambled FITC-PIF (control). Conclusions PIF is an early embryo viability marker that has a direct supportive role on embryo development in culture. PIF-ELISA use to assess IVF embryo quality prior to transfer is warranted. Overall, our data supports PIF's endogenous self sustaining role in embryo development and the utility of PIF- ELISA to detect viable embryos in a non-invasive manner. PMID:21569635

Improvement of aromatic thiol release through the selection of yeasts with increased β-lyase activity.

PubMed

Belda, Ignacio; Ruiz, Javier; Navascués, Eva; Marquina, Domingo; Santos, Antonio

2016-05-16

The development of a selective medium for the rapid differentiation of yeast species with increased aromatic thiol release activity has been achieved. The selective medium was based on the addition of S-methyl-l-cysteine (SMC) as β-lyase substrate. In this study, a panel of 245 strains of Saccharomyces cerevisiae strains was tested for their ability to grow on YCB-SMC medium. Yeast strains with an increased β-lyase activity grew rapidly because of their ability to release ammonium from SMC in comparison to others, and allowed for the easy isolation and differentiation of yeasts with promising properties in oenology, or another field, for aromatic thiol release. The selective medium was also helpful for the discrimination between those S. cerevisiae strains, which present a common 38-bp deletion in the IRC7 sequence (present in around 88% of the wild strains tested and are likely to be less functional for 4-mercapto-4-methylpentan-2-one (4MMP) production), and those S. cerevisiae strains homozygous for the full-length IRC7 allele. The medium was also helpful for the selection of non-Saccharomyces yeasts with increased β-lyase activity. Based on the same medium, a highly sensitive, reproducible and non-expensive GC-MS method for the evaluation of the potential volatile thiol release by different yeast isolates was developed. Copyright © 2016 Elsevier B.V. All rights reserved.

An incubation medium for the elevation of adenosine triphosphate and 2,3-diphosphoglycerate in fresh and long-preserved human erythrocytes.

PubMed

Rubinstein, D; Warrendorf, E

1975-06-01

The levels of adenosine triphosphate (ATP) and 2,3-diphosphoglycerate in freshly drawn human erythrocytes can be tripled by a 2 h incubation at 37 degrees C in a medium containing 21 mM glucose, 1.8 mM adenine, 5 mM pyruvate, 10 mM inosine, and 96 mM phosphate. Similar incubation conditions will restore the levels of ATP and 2,3-diphosphoglycerate in erythrocytes from blood levels preserved for 12 and 15 weeks, respectively, to those of fresh cells. Omission of pyruvate from the incubation medium further increases the level of ATP slightly, but there is little elevation of 2,3-diphosphoglycerate. Under these conditions labelled pyruvate and lactate production from [14-C]glucose or [14-C]inosine is not diminished, but labelled fructose 1,6-diphosphate, rather than 2,3-diphosphoglycerate, accumulates. In addition, omission of pyruvate from the incubation medium, with a concomitant decrease in accumulation of 2,3-diphosphoglycerate, diminishes the concentration of inorganic phosphate required for optimal ATP elevation. A 5 h incubation in the glucose-adenine-pyruvate-inosine-phosphate medium elevates the levels of ATP and 2,3-diphosphoglycerate in erythrocytes from blood preserved in the cold for 15 weeks to twice that of fresh cells, indicating that the cells retain their metabolic potential even after prolonged storage at 2 degrees C. The medium may provide a method of rejuvenating 10-12 week cold-preserved erythrocytes for transfusion purposes, by a 1 h incubation at 37 degrees C.

Targeting arginase-II protects mice from high-fat-diet-induced hepatic steatosis through suppression of macrophage inflammation.

PubMed

Liu, Chang; Rajapakse, Angana G; Riedo, Erwin; Fellay, Benoit; Bernhard, Marie-Claire; Montani, Jean-Pierre; Yang, Zhihong; Ming, Xiu-Fen

2016-02-05

Nonalcoholic fatty liver disease (NAFLD) associates with obesity and type 2 diabetes. Hypoactive AMP-activated protein kinase (AMPK), hyperactive mammalian target of rapamycin (mTOR) signaling, and macrophage-mediated inflammation are mechanistically linked to NAFLD. Studies investigating roles of arginase particularly the extrahepatic isoform arginase-II (Arg-II) in obesity-associated NAFLD showed contradictory results. Here we demonstrate that Arg-II(-/-) mice reveal decreased hepatic steatosis, macrophage infiltration, TNF-α and IL-6 as compared to the wild type (WT) littermates fed high fat diet (HFD). A higher AMPK activation (no difference in mTOR signaling), lower levels of lipogenic transcription factor SREBP-1c and activity/expression of lipogenic enzymes were observed in the Arg-II(-/-) mice liver. Moreover, release of TNF-α and IL-6 from bone marrow-derived macrophages (BMM) of Arg-II(-/-) mice is decreased as compared to WT-BMM. Conditioned medium from Arg-II(-/-)-BMM exhibits weaker activity to facilitate triglyceride synthesis paralleled with lower expression of SREBP-1c and SCD-1 and higher AMPK activation in hepatocytes as compared to that from WT-BMM. These effects of BMM conditioned medium can be neutralized by neutralizing antibodies against TNF-α and IL-6. Thus, Arg-II-expressing macrophages facilitate diet-induced NAFLD through TNF-α and IL-6 in obesity.

78 FR 72584 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-03

... medium or giant BFT per vessel per day/trip to five large medium or giant BFT per vessel per day/trip for... through December 31, 2013. The General category daily retention limit is currently three large medium or... category limit from the default level of one large medium or giant BFT for the 2013 January subquota period...

78 FR 77362 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-23

... large medium or giant BFT to two large medium or giant BFT for the January 2014 subquota period ((i.e... starting on January 1 would be the default retention limit of one large medium or giant BFT (measuring 73... adjusted the General category limit from the default level of one large medium or giant BFT to two large...

76 FR 76900 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

Federal Register 2010, 2011, 2012, 2013, 2014

2011-12-09

... retention limit of one large medium or giant BFT (measuring 73 inches (185 cm) CFL) or greater per vessel... level of one large medium or giant BFT as follows: Two large medium or giant BFT for January (75 FR 79309, December 20, 2010); three large medium or giant BFT for June through August (76 FR 32086, June 3...

75 FR 79309 - Atlantic Highly Migratory Species; Atlantic Bluefin Tuna Fisheries

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-20

... limit of one large medium or giant BFT (measuring 73 inches (185 cm) CFL) or greater per vessel per day... adjusted the General category limit from the default level of one large medium or giant BFT as follows: Two large medium or giant BFT for January (74 FR 68709, December 29, 2009), and three large medium or giant...

Chemical and genetic blockade of HDACs enhances osteogenic differentiation of human adipose tissue-derived stem cells by oppositely affecting osteogenic and adipogenic transcription factors

DOE Office of Scientific and Technical Information (OSTI.GOV)

Maroni, Paola; Brini, Anna Teresa; Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, Universita degli Studi di Milano, Milano

2012-11-16

Highlights: Black-Right-Pointing-Pointer Acetylation affected hASCs osteodifferentiation through Runx2-PPAR{gamma}. Black-Right-Pointing-Pointer HDACs knocking-down favoured the commitment effect of osteogenic medium. Black-Right-Pointing-Pointer HDACs silencing early activated Runx2 and ALP. Black-Right-Pointing-Pointer PPAR{gamma} reduction and calcium/collagen deposition occurred later. Black-Right-Pointing-Pointer Runx2/PPAR{gamma} target genes were modulated in line with HDACs role in osteo-commitment. -- Abstract: The human adipose-tissue derived stem/stromal cells (hASCs) are an interesting source for bone-tissue engineering applications. Our aim was to clarify in hASCs the role of acetylation in the control of Runt-related transcription factor 2 (Runx2) and Peroxisome proliferator activated receptor (PPAR) {gamma}. These key osteogenic and adipogenic transcription factors are oppositelymore » involved in osteo-differentiation. The hASCs, committed or not towards bone lineage with osteoinductive medium, were exposed to HDACs chemical blockade with Trichostatin A (TSA) or were genetically silenced for HDACs. Alkaline phosphatase (ALP) and collagen/calcium deposition, considered as early and late osteogenic markers, were evaluated concomitantly as index of osteo-differentiation. TSA pretreatment, useful experimental protocol to analyse pan-HDAC-chemical inhibition, and switch to osteogenic medium induced early-osteoblast maturation gene Runx2, while transiently decreased PPAR{gamma} and scarcely affected late-differentiation markers. Time-dependent effects were observed after knocking-down of HDAC1 and 3: Runx2 and ALP underwent early activation, followed by late-osteogenic markers increase and by PPAR{gamma}/ALP activity diminutions mostly after HDAC3 silencing. HDAC1 and 3 genetic blockade increased and decreased Runx2 and PPAR{gamma} target genes, respectively. Noteworthy, HDACs knocking-down favoured the commitment effect of osteogenic medium. Our results reveal a role for HDACs in orchestrating osteo-differentiation of hASCs at transcriptional level, and might provide new insights into the modulation of hASCs-based regenerative therapy.« less

Phytase-producing capacity of yeasts isolated from traditional African fermented food products and PHYPk gene expression of Pichia kudriavzevii strains.

PubMed

Greppi, Anna; Krych, Łukasz; Costantini, Antonella; Rantsiou, Kalliopi; Hounhouigan, D Joseph; Arneborg, Nils; Cocolin, Luca; Jespersen, Lene

2015-07-16

Phytate is known as a strong chelate of minerals causing their reduced uptake by the human intestine. Ninety-three yeast isolates from traditional African fermented food products, belonging to nine species (Pichia kudriavzevii, Saccharomyces cerevisiae, Clavispora lusitaniae, Kluyveromyces marxianus, Millerozyma farinosa, Candida glabrata, Wickerhamomyces anomalus, Hanseniaspora guilliermondii and Debaryomyces nepalensis) were screened for phytase production on solid and liquid media. 95% were able to grow in the presence of phytate as sole phosphate source, P. kudriavzevii being the best growing species. A phytase coding gene of P. kudriavzevii (PHYPk) was identified and its expression was studied during growth by RT-qPCR. The expression level of PHYPk was significantly higher in phytate-medium, compared to phosphate-medium. In phytate-medium expression was seen in the lag phase. Significant differences in gene expression were detected among the strains as well as between the media. A correlation was found between the PHYPk expression and phytase extracellular activity. Copyright © 2015 Elsevier B.V. All rights reserved.

Pentagalloyl glucose increases elastin deposition, decreases reactive oxygen species and matrix metalloproteinase activity in pulmonary fibroblasts under inflammatory conditions.

PubMed

Parasaram, Vaideesh; Nosoudi, Nasim; Chowdhury, Aniqa; Vyavahare, Naren

2018-04-30

Emphysema is characterized by degradation of lung alveoli that leads to poor airflow in lungs. Irreversible elastic fiber degradation by matrix metalloproteinases (MMPs) and reactive oxygen species (ROS) activity leads to loss of elasticity and drives the progression of this disease. We investigated if a polyphenol, pentagalloyl glucose (PGG) can increase elastin production in pulmonary fibroblasts. We also studied the effect of PGG treatment in reducing MMP activity and ROS levels in cells. We exposed rat pulmonary fibroblasts to two different types of inflammatory environments i.e., tumor necrosis factor-α (TNF-α) and cigarette smoke extract (CSE) to mimic the disease. Parameters like lysyl oxidase (LOX) and elastin gene expression, MMP-9 activity in the medium, lysyl oxidase (LOX) activity and ROS levels were studied to assess the effect of PGG on pulmonary fibroblasts. CSE inhibited lysyl oxidase (LOX) enzyme activity that resulted in a decreased elastin formation. Similarly, TNF-α treated cells showed less elastin in the cell layers. Both these agents caused increase in MMP activity and ROS levels in cells. However, when supplemented with PGG treatment along with these two inflammatory agents, we saw a significant increase in elastin deposition, reduction in both MMP activity and ROS levels. Thus PGG, which has anti-inflammatory, anti-oxidant properties coupled with its ability to aid in elastic fiber formation, can be a multifunctional drug to potentially arrest the progression of emphysema. Copyright © 2018 Elsevier Inc. All rights reserved.

Energy and calcium ion dependence of proteolysis during sporulation of Bacillus subtilis cells.

PubMed

O'Hara, M B; Hageman, J H

1990-08-01

Bacterial cells degrade intracellular proteins at elevated rates during starvation and can selectively degrade proteins by energy-dependent processes. Sporulating bacteria can degrade protein with apparent first-order rate constants of over 0.20 h-1. We have shown, with an optimized [14C]leucine-labeling and chasing procedure, in a chemically defined sporulation medium, that intracellular protein degradation in sporulating cells of Bacillus subtilis 168 (trpC2) is apparently energy dependent. Sodium arsenate, sodium azide, carbonyl cyanide m-chlorophenylhydrozone, and N,N'-dicyclohexylcarbodiimide, at levels which did not induce appreciable lysis (less than or equal to 10%) over 10-h periods of sporulation, inhibited intracellular proteolysis by 13 to 93%. Exponentially growing cells acquired arsenate resistance. In contrast to earlier reports, we found that chloramphenicol (100 micrograms/ml) strongly inhibited proteolysis (68%) even when added 6 h into the sporulation process. Restricting the calcium ion concentration (less than 2 microM) in the medium had no effect on rates or extent of vegetative growth, strongly inhibited sporulation (98%), and inhibited rates of proteolysis by 60% or more. Inhibitors of energy metabolism, at the same levels which inhibited proteolysis, did not affect the rate or degree of uptake of Ca2+ by cells, which suggested that the Ca2+ and metabolic energy requirements of proteolysis were independent. Restricting the Ca2+ concentration in the medium reduced by threefold the specific activity in cells of the major intracellular serine proteinase after 12 h of sporulation. Finally, cells of a mutant of B. subtilis bearing an insertionally inactivated gene for the Ca2(+)-dependent intracellular proteinase-1 degraded protein in chemically defined sporulation medium at a rate indistinguishable from that of the wild-type cells for periods of 8 h.

Betel Nut Chewing Is Associated With Reduced Tacrolimus Concentration in Taiwanese Liver Transplant Recipients.

PubMed

Chen, W-Y; Lee, C-Y; Lin, P-Y; Hsieh, C-E; Ko, C-J; Lin, K-H; Lin, C-C; Ming, Y-Z; Chen, Y-L

2017-03-01

Studies have shown that arecoline, the major alkaloid component of betel nuts, alters the activity of enzymes in the cytochrome P450 (CYP-450) family. Tacrolimus, an immunosuppressant that protects against organ rejection in transplant recipients, not only is mainly metabolized by CYP3A enzymes but also has a narrow therapeutic range. We aimed to investigate whether dose-adjusted blood trough levels of tacrolimus differed over time between betel nut-chewing and non-betel nut-chewing liver transplant recipients. In this retrospective case-control study, 14 active betel nut-using liver recipients were matched at a 1:2 ratio to 28 non-betel nut-using liver recipients by sex, age, graft source, duration of follow-up after liver transplantation, and estimated glomerular filtration rate. Differences in liver function index, renal function index, and dose-adjusted blood trough levels of tacrolimus over an 18-month period were compared between the 2 groups by using the Generalized Estimating Equation approach. Dose-adjusted blood trough levels of tacrolimus tended to be significantly (P = .04) lower in betel nut chewers (mean = 0.81, medium = 0.7, 95% confidence interval [CI] = 0.73 to 0.90) than in nonchewers (mean = 1.12, medium = 0.88, 95% CI = 1.03 to 1.22) during the 18-month study period. However, there was no significant difference in renal and liver function index between the 2 groups. Liver transplant recipients receiving tacrolimus tend to have lower blood trough levels of the drug over time if they chew betel nuts. Copyright © 2016 Elsevier Inc. All rights reserved.

Successful return to sports in athletes following non-operative management of acute isolated posterior cruciate ligament injuries: medium-term follow-up.

PubMed

Agolley, D; Gabr, A; Benjamin-Laing, H; Haddad, F S

2017-06-01

The aim of this study was to report the outcome of the non-operative treatment of high-grade posterior cruciate ligament (PCL) injuries, particularly Hughston grade III injuries, which have not previously been described. This was a prospective study involving 46 consecutive patients who were athletes with MRI-confirmed isolated PCL injuries presenting within four weeks of injury. All had Hughston grade II (25 athletes) or III (21 athletes) injuries. Our non-operative treatment regimen involved initial bracing, followed by an individualised rehabilitation programme determined by the symptoms and physical signs. The patients were reviewed until they had returned to sports-specific training, and were reviewed again at a mean of 5.2 years (3 to 9). The mean time to return to sports-specific training was 10.6 weeks and the mean time to return to full competitive sport was 16.4 weeks (10 to 40). A total of 42 patients (91.3%) were playing at the same or higher level of sport two years after the injury, with a mean Tegner activity score of 9 (5 to 10). At five years, 32 patients (69.5%) were playing at the same or higher level of sport, and 38 patients (82.6%) were playing at a competitive level, with a mean Tegner activity score of 9 (5 to 10). Medium-term review of a series of athletes suggests that commencing the non-operative management of isolated, Hughston grade II and III PCL injuries within four weeks of injury gives excellent functional outcomes with a high proportion returning to the same or higher level of sport. Cite this article: Bone Joint J 2017;99-B:774-8. ©2017 The British Editorial Society of Bone & Joint Surgery.

[Problems of implementing integration management at company level in small and medium-sized enterprises].

PubMed

Hetzel, C; Flach, T; Weber, A; Schian, H-M

2006-05-01

At company level responsibility increases for the employment of workers with health-related problems or disabilities, but realisation in small and medium-sized enterprises (SME) is lacking. Therefore a model is developed based on theory and a survey. Minimum requirements for "betriebliches Eingliederungsmanagement" (integration management at company level) according to section 84 (2) SGB IX Book 9 of the German Social Code, the main products of the international movement "disability management", a description of roles for realisation and the main sources of employers' support are described. Although external supporting of SMEs is unquestioned, it is expensive and retards own initiative and own activity counting solely on this. Only by developing a minimum of SME's awareness, acceptance and competence, this will open up to (currently suboptimal) external support. Goal is identifying SME managers' attitudes, activities, proposals and expectations referring integration management at company level to derive concepts of SME's support. 13 interviews are analysed by qualitative content analysis identifying the following barriers: information deficit, absence of priority, limited possibilities for transitional work, cost, partially limited workers' self-responsibility, illness as a "tabes" subject. Possibilities overcoming these barriers are delineated. On that basis a model is presented: pragmatically for realisation, a company contact person with minimum competence, uniform external support, institutional partners' integration and quality assurance according to disability management. Interlocking SME world and social insurance world means first to support SME's awareness, acceptance and competence, second to create for SME a central contact in the "rehabilitation jungle" and third to develop SME-suitable premiums according to section 84 (3) SGB IX, Book 9 of the German social code.

Modulation of the Effect of l‐β‐D‐Arabinofuranosylcytosine by 6‐Mercaptopurine in L1210 Cells

PubMed Central

Kawai, Yasukazu; Ueda, Takanori; Nakamura, Toru

1994-01-01

In L1210 cells incubated with l‐β‐D‐arabinofuranosylcytosine (ara‐C), 6‐mercaptopurine (6‐MP) significantly potentiated 1‐β‐D‐arabinofuranosylcytosme 5′‐triphosphate (ara‐CTP) accumulation and ara‐C incorporation into DNA (ara‐C/DNA). The cytotoxicity of these two drugs was assessed to be at least additive by clonogenic assay. l‐β‐D‐Arabinofuranosylnracil (ara‐U) level in a cell suspension was suppressed by 6‐MP in a concentration‐dependent fashion, though intracellular cytidine deaminase (CDD) activity was not affected by 6‐MP. In addition, extracted CDD activity was not directly inhibited by 6‐MP or by its intracellular metabolites in vitro. After preincubation in the presence or absence of 6‐MP, the cell suspension was fractionated to obtain the spent medium and cell pellet. Then, each fraction was incubated with ara‐C. Ara‐U formation in the spent medium was found to increase conspicuously in relation to the time of preincubation in the control and it was suppressed by 6‐MP pretreatment. Ara‐U formation in the cell compartment increased slightly in relation to the time of preincubation in the control and substantially no suppression of ara‐U formation was observed in spite of 6‐MP pretreatment. In conclusion, intracellularly synthesized CDD was thought to be rapidly shed into the medium and the released CDD could play an important role in ara‐C inactivation. 6‐MP interrupted some step between synthesis and shedding of CDD, resulting in a decrease of the ara‐C deamination in the medium and enhancement of its antileukemic effect. PMID:7961129

Enhancement of Penicillium echinulatum glycoside hydrolase enzyme complex.

PubMed

dos Santos Costa, Patrícia; Büchli, Fernanda; Robl, Diogo; Delabona, Priscila da Silva; Rabelo, Sarita Candida; Pradella, José Geraldo da Cruz

2016-05-01

The enhancement of enzyme complex produced by Penicillium echinulatum grown in several culture media components (bagasse sugarcane pretreated by various methods, soybean meal, wheat bran, sucrose, and yeast extract) was studied to increment FPase, xylanase, pectinase, and β-glucosidase enzyme activities. The present results indicated that culture media composed with 10 g/L of the various bagasse pretreatment methods did not have any substantial influence with respect to the FPase, xylanase, and β-glucosidase attained maximum values of, respectively, 2.68 FPU/mL, 2.04, and 115.4 IU/mL. On the other hand, proposed culture media to enhance β-glucosidase production composed of 10 g/L steam-exploded bagasse supplemented with soybean flour 5.0 g/L, yeast extract 1.0 g/L, and sucrose 10.0 g/L attained, respectively, 3.19 FPU/mL and 3.06 IU/mL while xylanase was maintained at the same level. The proteomes obtained from the optimized culture media for enhanced FPase, xylanase, pectinase, and β-glucosidase production were analyzed using mass spectrometry and a panel of GH enzyme activities against 16 different substrates. Culture medium designed to enhance β-glucosidase activity achieved higher enzymatic activities values (13 measured activities), compared to the culture media for FPase/pectinase (9 measured activities) and xylanase (7 measured activities), when tested against the 16 substrates. Mass spectrometry analyses of secretome showed a consistent result and the greatest number of spectral counts of Cazy family enzymes was found in designed β-glucosidase culture medium, followed by FPase/pectinase and xylanase. Most of the Cazy identified protein was cellobiohydrolase (GH6 and GH7), endoglucanase (GH5), and endo-1,4-β-xylanase (GH10). Enzymatic hydrolysis of hydrothermally pretreated sugarcane bagasse performed with β-glucosidase enhanced cocktail achieved 51.4 % glucose yield with 10 % w/v insoluble solids at enzyme load of 15 FPU/g material. Collectively the results demonstrated that it was possible to rationally modulate the GH activity of the enzymatic complex secreted by P. echinulatum using adjustment of the culture medium composition. The proposed strategy may contribute to increase enzymatic hydrolysis of lignocellulosic materials.

Use of the short-term inflammatory response in the mouse peritoneal cavity to assess the biological activity of leached vitreous fibers.

PubMed Central

Donaldson, K; Addison, J; Miller, B G; Cullen, R T; Davis, J M

1994-01-01

We used a special-purpose glass microfiber sample, Johns-Manville Code 100/475, to study the effects of various acid and alkali treatments on biological activity as assessed by inflammation in the mouse peritoneal cavity, the leaching of Si, and the phase contrast optical microscopy (PCOM) fiber number. We used mild and medium treatments with oxalic acid and Tris buffer and harsh treatment with concentrated HCl and NaOH. Mild oxalic acid and Tris treatment for 2 weeks had no effect on any of the end-points, but prolonging the mild oxalic acid treatment time to 2 months reduced the biological activity and the fiber number. Medium oxalic acid treatment reduced the biological activity and the fiber number and caused a loss of Si. Medium Tris alkali treatment reduced the PCOM-countable fibers and the biological activity but did not cause a substantial loss of Si. Harsh treatment with strong HCl did not affect the fiber number or cause leaching but the biological activity was reduced; strong NaOH reduced the fiber number and biological activity, and caused marked leaching of Si. The medium oxalic acid conditions (pH 1.4) were more acid than those found in lung cells but produced the same effects (reduction in fiber number and biological activity) as the more physiological mild treatment (pH 4.0), when prolonged. This study suggests that medium oxalic acid treatment can be used as a short-term assay to compare loss of Si, reduction in fiber number, and change in biological activity of vitreous fibers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7882922

Polycystin-1 is a Cardiomyocyte Mechanosensor That Governs L-type Ca2+ Channel Protein Stability

PubMed Central

Pedrozo, Zully; Criollo, Alfredo; Battiprolu, Pavan K.; Morales, Cyndi R.; Contreras, Ariel; Fernández, Carolina; Jiang, Nan; Luo, Xiang; Caplan, Michael J.; Somlo, Stefan; Rothermel, Beverly A.; Gillette, Thomas G.; Lavandero, Sergio; Hill, Joseph A.

2015-01-01

Background L-type calcium channel (LTCC) activity is critical to afterload-induced hypertrophic growth of the heart. However, mechanisms governing mechanical stress-induced activation of LTCC activity are obscure. Polycystin-1 (PC-1) is a G-protein-coupled receptor-like protein that functions as a mechanosensor in a variety of cell types and is present in cardiomyocytes. Methods and Results We subjected neonatal rat ventricular myocytes (NRVMs) to mechanical stretch by exposing them to hypo-osmotic (HS) medium or cyclic mechanical stretch, triggering cell growth in a manner dependent on LTCC activity. RNAi-dependent knockdown of PC-1 blocked this hypertrophy. Over-expression of a C-terminal fragment of PC-1 was sufficient to trigger NRVM hypertrophy. Exposing NRVMs to HS medium resulted in an increase in α1C protein levels, a response that was prevented by PC-1 knockdown. MG132, a proteasomal inhibitor, rescued PC-1 knockdown-dependent declines in α1C protein. To test this in vivo, we engineered mice harboring conditional silencing of PC-1 selectively in cardiomyocytes (PC-1 KO) and subjected them to mechanical stress in vivo (transverse aortic constriction, TAC). At baseline, PC-1 KO mice manifested decreased cardiac function relative to littermate controls, and α1C LTCC protein levels were significantly lower in PC-1 KO hearts. Whereas control mice manifested robust TAC-induced increases in cardiac mass, PC-1 KO mice showed no significant growth. Likewise, TAC-elicited increases in hypertrophic markers and interstitial fibrosis were blunted in the knockout animals Conclusions PC-1 is a cardiomyocyte mechanosensor and is required for cardiac hypertrophy through a mechanism that involves stabilization of α1C protein. PMID:25888683

Polycystin-1 Is a Cardiomyocyte Mechanosensor That Governs L-Type Ca2+ Channel Protein Stability.

PubMed

Pedrozo, Zully; Criollo, Alfredo; Battiprolu, Pavan K; Morales, Cyndi R; Contreras-Ferrat, Ariel; Fernández, Carolina; Jiang, Nan; Luo, Xiang; Caplan, Michael J; Somlo, Stefan; Rothermel, Beverly A; Gillette, Thomas G; Lavandero, Sergio; Hill, Joseph A

2015-06-16

L-type calcium channel activity is critical to afterload-induced hypertrophic growth of the heart. However, the mechanisms governing mechanical stress-induced activation of L-type calcium channel activity are obscure. Polycystin-1 (PC-1) is a G protein-coupled receptor-like protein that functions as a mechanosensor in a variety of cell types and is present in cardiomyocytes. We subjected neonatal rat ventricular myocytes to mechanical stretch by exposing them to hypo-osmotic medium or cyclic mechanical stretch, triggering cell growth in a manner dependent on L-type calcium channel activity. RNAi-dependent knockdown of PC-1 blocked this hypertrophy. Overexpression of a C-terminal fragment of PC-1 was sufficient to trigger neonatal rat ventricular myocyte hypertrophy. Exposing neonatal rat ventricular myocytes to hypo-osmotic medium resulted in an increase in α1C protein levels, a response that was prevented by PC-1 knockdown. MG132, a proteasomal inhibitor, rescued PC-1 knockdown-dependent declines in α1C protein. To test this in vivo, we engineered mice harboring conditional silencing of PC-1 selectively in cardiomyocytes (PC-1 knockout) and subjected them to mechanical stress in vivo (transverse aortic constriction). At baseline, PC-1 knockout mice manifested decreased cardiac function relative to littermate controls, and α1C L-type calcium channel protein levels were significantly lower in PC-1 knockout hearts. Whereas control mice manifested robust transverse aortic constriction-induced increases in cardiac mass, PC-1 knockout mice showed no significant growth. Likewise, transverse aortic constriction-elicited increases in hypertrophic markers and interstitial fibrosis were blunted in the knockout animals PC-1 is a cardiomyocyte mechanosensor that is required for cardiac hypertrophy through a mechanism that involves stabilization of α1C protein. © 2015 American Heart Association, Inc.

Addition of bone morphogenetic protein type 2 to ascorbate and β-glycerophosphate supplementation did not enhance osteogenic differentiation of human adipose-derived stem cells.

PubMed

Cruz, Ariadne Cristiane Cabral; Silva, Mariana Lúcia; Caon, Thiago; Simões, Cláudia Maria Oliveira

2012-01-01

Bone morphogenetic protein type 2 (BMP-2) is a potent local factor, which promotes bone formation and has been used as an osteogenic supplement for mesenchymal stem cells. This study evaluated the effect of a recombinant BMP-2 as well as the endogenous BMP-4 and BMP-7 in the osteogenic differentiation of adipose-derived stem cells (ASCs) in medium supplemented with ascorbate and β-glycerophosphate. Human ASCs were treated with osteogenic medium in the presence (ASCs+OM+BMP-2) or absence (ASCs+OM) of BMP-2. The alkaline phosphatase (ALP) activity was determined and the extracellular matrix mineralization was evaluated by Von Kossa staining and calcium quantification. The expressions of BMP-4, BMP-7, Smad1, Smad4, and phosphorylated Smad1/5/8 were analyzed by western blotting. Relative mRNA expressions of Smad1, BMP receptor type II (BMPR-II), osteonectin, and osteocalcin were evaluated by qPCR. ASCs+OM demonstrated the highest expression of BMP-4 and BMP-7 at days 21 and 7, respectively, the highest levels of BMPR-II mRNA expression at day 28, and the highest levels of Smad1 mRNA at days 14 and 28. ASCs+OM+BMP-2 demonstrated the highest levels of Smad1 mRNA expression at days 1, 7, and 21, the highest expression of Smad1 at day 7, the highest expression of Smad4 at day 14, the highest ALP activity at days 14 and 21, and expression of phosphorylated Smad1/5/8 at day 7. ASCs+OM and ASCs+OM+BMP2 showed similar ALP activity at days 7 and 28, similar osteonectin and osteocalcin mRNA expression at all time periods, and similar calcium depositions at all time periods. We concluded that human ASCs expressed endogenous BMP-4 and BMP-7. Moreover, the supplementation of ASCs with BMP-2 did not increase the level of osteogenic markers in the initial (ALP activity), intermediate (osteonectin and osteocalcin), or final (calcium deposition) phases, suggesting that the exogenous addition of BMP-2 did not improve the in vitro osteogenesis process of human ASCs.

[Antibacterial activity of 20 kinds of Chinese medicinal materials for Helicobacter pylori in vitro].

PubMed

Du, P; Zhu, S; Lü, P

2001-03-01

To study the antibacterial activity of 20 Chinese medicinal materials for helicobacter pylori in vitro and the culture of hylicobacter pylori. Doubled-diluted method and tomato juice culture medium were adopted. The detecting ratio of tomato juice culture medium to hylicobacter pylori was equal to that of skirrow culture medium; Radix Scutellariae, Flos Lonicerae, Radix Isatidis, Indigo Naturalis, Fructus Chebulae, Semen Ginkgo, Cortex Phellodendri, Rhizoma Corydalis and Cortex Fraxini have obvious effect of antibacterium to hyliocobater pylori.

Gene expression responses of HeLa cells to chemical species generated by an atmospheric plasma flow

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yokoyama, Mayo, E-mail: yokoyama@plasma.ifs.tohoku.ac.jp; Johkura, Kohei, E-mail: kohei@shinshu-u.ac.jp; Sato, Takehiko, E-mail: sato@ifs.tohoku.ac.jp

2014-08-08

Highlights: • Response of HeLa cells to a plasma-irradiated medium was revealed by DNA microarray. • Gene expression pattern was basically different from that in a H{sub 2}O{sub 2}-added medium. • Prominently up-/down-regulated genes were partly shared by the two media. • Gene ontology analysis showed both similar and different responses in the two media. • Candidate genes involved in response to ROS were detected in each medium. - Abstract: Plasma irradiation generates many factors able to affect the cellular condition, and this feature has been studied for its application in the field of medicine. We previously reported that hydrogenmore » peroxide (H{sub 2}O{sub 2}) was the major cause of HeLa cell death among the chemical species generated by high level irradiation of a culture medium by atmospheric plasma. To assess the effect of plasma-induced factors on the response of live cells, HeLa cells were exposed to a medium irradiated by a non-lethal plasma flow level, and their gene expression was broadly analyzed by DNA microarray in comparison with that in a corresponding concentration of 51 μM H{sub 2}O{sub 2}. As a result, though the cell viability was sufficiently maintained at more than 90% in both cases, the plasma-medium had a greater impact on it than the H{sub 2}O{sub 2}-medium. Hierarchical clustering analysis revealed fundamentally different cellular responses between these two media. A larger population of genes was upregulated in the plasma-medium, whereas genes were downregulated in the H{sub 2}O{sub 2}-medium. However, a part of the genes that showed prominent differential expression was shared by them, including an immediate early gene ID2. In gene ontology analysis of upregulated genes, the plasma-medium showed more diverse ontologies than the H{sub 2}O{sub 2}-medium, whereas ontologies such as “response to stimulus” were common, and several genes corresponded to “response to reactive oxygen species.” Genes of AP-1 proteins, e.g., JUN and FOS, were detected and notably elevated in the plasma-medium. These results showed that the medium irradiated with a non-lethal level of plasma flow altered various gene expressions of HeLa cells by giving not only common effects with H{sub 2}O{sub 2} but also some distinctive actions. This study suggests that in addition to H{sub 2}O{sub 2}, other chemical species able to affect the cellular responses exist in the plasma-irradiated medium and provide unique features for it, probably increasing the oxidative stress level.« less

Evaluation of an active humidification system for inspired gas.

PubMed

Roux, Nicolás G; Plotnikow, Gustavo A; Villalba, Darío S; Gogniat, Emiliano; Feld, Vivivana; Ribero Vairo, Noelia; Sartore, Marisa; Bosso, Mauro; Scapellato, José L; Intile, Dante; Planells, Fernando; Noval, Diego; Buñirigo, Pablo; Jofré, Ricardo; Díaz Nielsen, Ernesto

2015-03-01

The effectiveness of the active humidification systems (AHS) in patients already weaned from mechanical ventilation and with an artificial airway has not been very well described. The objective of this study was to evaluate the performance of an AHS in chronically tracheostomized and spontaneously breathing patients. Measurements were quantified at three levels of temperature (T°) of the AHS: level I, low; level II, middle; and level III, high and at different flow levels (20 to 60 L/minute). Statistical analysis of repeated measurements was performed using analysis of variance and significance was set at a P<0.05. While the lowest temperature setting (level I) did not condition gas to the minimum recommended values for any of the flows that were used, the medium temperature setting (level II) only conditioned gas with flows of 20 and 30 L/minute. Finally, at the highest temperature setting (level III), every flow reached the minimum absolute humidity (AH) recommended of 30 mg/L. According to our results, to obtain appropiate relative humidity, AH and T° of gas one should have a device that maintains water T° at least at 53℃ for flows between 20 and 30 L/m, or at T° of 61℃ at any flow rate.

Critical thinking level in geometry based on self-regulated learning

NASA Astrophysics Data System (ADS)

Bayuningsih, A. S.; Usodo, B.; Subanti, S.

2018-03-01

Critical thinking ability of mathematics students affected by the student’s ability in solving a specific problem. This research aims to determine the level of critical thinking (LCT) students in solving problems of geometry regarding self-regulated learning (SRL) students. This is a qualitative descriptive study with the purpose to analyze the level of Junior High School student’s critical thinking in the Regency of Banyumas. The subject is taken one student from each category SRL (high, medium and low). Data collection is given problem-solving tests to find out the level of critical thinking student, questionnaire, interview and documentation. The result of the research shows that student with SRL high is at the level of critical thinking 2, then a student with SRL medium is at the level of critical thinking 1 and student with SRL low is at the level of critical thinking 0. So students with SRL high, medium or low can solve math problems based on the critical thinking level of each student.

Inhibition effects of chlorogenic acid on benign prostatic hyperplasia in mice.

PubMed

Huang, Ya; Chen, Huaguo; Zhou, Xin; Wu, Xingdong; Hu, Enming; Jiang, Zhengmeng

2017-08-15

This study aimed to evaluate the inhibitory effects and explore mechanisms of chlorogenic acid against testosterone-induced benign prostatic hyperplasia (BPH) in mice. Benign prostatic hyperplasia model was induced in experimental groups by daily subcutaneous injections of testosterone propionate (7.5mg/kg/d) consecutively for 14 d. A total of 60 mice were randomly divided into six groups: (Group 1) normal control group, (Group 2) benign prostatic hyperplasia model control group, (Group 3) benign prostatic hyperplasia mice treated with finasteride at a dose of 1mg/kg, (Group 4) benign prostatic hyperplasia mice treated with chlorogenic acid at dose levels of 0.8mg/kg (low dose group), (Group 5) benign prostatic hyperplasia mice treated with chlorogenic acid at dose levels of 1.6mg/kg (medium dose group) and (Group 6) benign prostatic hyperplasia mice treated with chlorogenic acid at dose levels of 3.2mg/kg (high dose group). Animals were sacrificed on the scheduled termination, pick out the eyeball to get blood, then prostates were weighed and prostatic index were determined. Then the serum acid phosphatase (ACP), prostatic acid phosphatase (PACP) and typeⅡ5-alpha-reductase (SRD5A2) levels were measured and observed morphological changes of the prostate. Comparing with benign prostatic hyperplasia model group, the high and medium dose of chlorogenic acid could significantly reduce prostate index and levels of acid phosphatase, prostatic acid phosphatase and typeⅡ5-alpha-reductase (P<0.05 or P<0.01). These findings were supported by histopathological observations of prostate tissues. Histopathological examination also indicated that chlorogenic acid treatment at the high and medium doses inhibited testosterone-induced prostatic hyperplasia. The results indicated that chlorogenic acid exhibited restraining effect on benign prostatic hyperplasia model animals, and its mechanism might be related to inhibit typeⅡ5-alpha reductase activity. Copyright © 2017. Published by Elsevier B.V.

Physical activity and unplanned illness-related work absenteeism: Data from an employee wellness program.

PubMed

Losina, Elena; Yang, Heidi Y; Deshpande, Bhushan R; Katz, Jeffrey N; Collins, Jamie E

2017-01-01

Illness-related absenteeism is a major threat to work productivity. Our objective was to assess the relationship between physical activity and unplanned illness-related absenteeism from work. We implemented physical activity program for sedentary non-clinician employees of a tertiary medical center. Financial rewards were available for reaching accelerometer-measured ambulatory physical activity goals over a 24-week period. We categorized participants into three groups based on mean levels of physical activity: low (0-74 min/week), medium (75-149 min/week) and meeting CDC guidelines (≥150 min/week). We built a multivariable Poisson regression model to evaluate the relationship between physical activity and rates of unplanned illness-related absenteeism. The sample consisted of 292 employees who participated in the program. Their mean age was 38 years (SD 11), 83% were female, and 38% were obese. Over the 24 intervention weeks, participants engaged in a mean of 90 min/week (SD 74) of physical activity and missed a mean of 14 hours of work (SD 38) due to illness. Unplanned absenteeism due to illness was associated with physical activity. As compared to the group meeting CDC guidelines, in multivariable analyses those in the medium physical activity group had a 2.4 (95% CI 1.3-4.5) fold higher rate of illness-related absenteeism and those in the lowest physical activity group had a 3.5 (95% CI 1.7-7.2) fold higher rate of illness-related absenteeism. Less physical activity was associated with more illness-related absenteeism. Workforce-based interventions to increase physical activity may thus be a promising vehicle to reduce unplanned illness-related absenteeism.

Physical activity and unplanned illness-related work absenteeism: Data from an employee wellness program

PubMed Central

Yang, Heidi Y.; Katz, Jeffrey N.; Collins, Jamie E.

2017-01-01

Background Illness-related absenteeism is a major threat to work productivity. Our objective was to assess the relationship between physical activity and unplanned illness-related absenteeism from work. Methods We implemented physical activity program for sedentary non-clinician employees of a tertiary medical center. Financial rewards were available for reaching accelerometer-measured ambulatory physical activity goals over a 24-week period. We categorized participants into three groups based on mean levels of physical activity: low (0–74 min/week), medium (75–149 min/week) and meeting CDC guidelines (≥150 min/week). We built a multivariable Poisson regression model to evaluate the relationship between physical activity and rates of unplanned illness-related absenteeism. Results The sample consisted of 292 employees who participated in the program. Their mean age was 38 years (SD 11), 83% were female, and 38% were obese. Over the 24 intervention weeks, participants engaged in a mean of 90 min/week (SD 74) of physical activity and missed a mean of 14 hours of work (SD 38) due to illness. Unplanned absenteeism due to illness was associated with physical activity. As compared to the group meeting CDC guidelines, in multivariable analyses those in the medium physical activity group had a 2.4 (95% CI 1.3–4.5) fold higher rate of illness-related absenteeism and those in the lowest physical activity group had a 3.5 (95% CI 1.7–7.2) fold higher rate of illness-related absenteeism. Discussion Less physical activity was associated with more illness-related absenteeism. Workforce-based interventions to increase physical activity may thus be a promising vehicle to reduce unplanned illness-related absenteeism. PMID:28472084

Influence of medium components on the expression of recombinant lipoproteins in Escherichia coli.

PubMed

Tseng, Chi-Ling; Leng, Chih-Hsiang

2012-02-01

Bacterial lipoproteins are crucial antigens for protective immunity against bacterial pathogens. Expression of exogenous lipoproteins in Escherichia coli at high levels is thought to be an extremely difficult endeavor because it frequently results in incomplete or absent lipid modification. Previously, we identified a fusion sequence (D1) from a Neisseria meningitidis lipoprotein that induced a non-lipidated protein, E3 (the domain III of the dengue virus envelope protein), to become lipidated. However, without optimizing the growth conditions, some of the D1-fusion proteins were not lipidated. Here, we report the influence of medium components on the expression of recombinant lipoproteins in E. coli. For high-level expression of mature lipoproteins in the C43 (DE3) strain, M9 medium was better than M63 and the rich medium. Furthermore, we analyzed the influence of other media factors (including nitrogen and carbon sources, phosphate, ferrous ions, calcium, magnesium, and pH) on the levels of lipoprotein expression. The results showed that excess nitrogen sources and phosphate in M9 medium could increase the amount of immature lipoproteins, and glucose was a better carbon source than glycerol for expressing mature lipoproteins. We also found that lipoproteins tended to be completely processed in the alkaline environment, even in the nutrient-rich medium. Additional constructs expressing different immunogens or lipid signal peptides as targets were also utilized, demonstrating that these targets could be expressed as completely mature lipoproteins in the M9 medium but not in the rich medium. Our results provide the useful information for expressing mature exogenous lipoproteins in E. coli.

Degradation of the herbicide paraquat by macromycetes isolated from southeastern Mexico.

PubMed

Camacho-Morales, Reyna L; Guillén-Navarro, Karina; Sánchez, José E

2017-10-01

Fifty-four macromycetes, isolated from southeastern Mexico, were used in order to evaluate their capacity for degradation and tolerance to the herbicide paraquat. Ten of these strains were capable of growing in a solid culture medium in the presence of 200 ppm paraquat. Subsequently, assays to evaluate the degradation of the xenobiotic in a liquid medium were carried out. Of the ten strains evaluated, three presented the highest levels of degradation of the compound, which were Trametes pavonia (54.2%), Trametes versicolor (54.1%) and Hypholoma dispersum. They presented the highest overall degradation percentage (70.7%) after 12 days culture. The presence of ligninolytic enzymes in these strains was evaluated. H. dispersum only presented aryl alcohol oxidase activity; however, with the data obtained, it was not possible to conclude whether this specific enzyme is responsible for paraquat degradation. The level of degradation obtained is above the one reported for Pseudomonas putida , one of the few reports on paraquat degradation. This is the first report on the contaminant degradation capacity of H. dispersum .

Medium Chain Triglycerides enhances exercise endurance through the increased mitochondrial biogenesis and metabolism.

PubMed

Wang, Ying; Liu, Zhenzhen; Han, Yi; Xu, Jiping; Huang, Wen; Li, Zhaoshen

2018-01-01

Medium Chain Triglycerides (MCT) is a dietary supplement and usually used along with medications for treating food absorption disorders including diarrhea, steatorrhea and liver disease. It has been shown that MCT plays a role in lowering weight, and decreasing metabolic syndrome, abdominal obesity and inflammation. However, it is still unknown whether MCT enhances exercise endurance. Here, we demonstrated that MCT containing diet improves high temperature induced exercise performance impairment. We found that MCT up-regulates the expression and protein levels of genes involved in mitochondrial biogenesis and metabolism. Further investigation demonstrated that the increased mitochondrial biogenesis and metabolism is mediated through the activation of Akt and AMPK signaling pathways and inhibition of TGF-β signaling pathway. Collectively, our findings indicate a beneficial effect of dietary MCT in exercise performance through the increase of mitochondrial biogenesis and metabolism.

Chromosome movement in lysed mitotic cells is inhibited by vanadate

PubMed Central

1978-01-01

Mitotic PtK1 cells, lysed at anaphase into a carbowax 20 M Brij 58 solution, continue to move chromosomes toward the spindle poles and to move the spindle poles apart at 50% in vivo rates for 10 min. Chromosome movements can be blocked by adding metabolic inhibitors to the lysis medium and inhibition of movement can be reversed by adding ATP to the medium. Vanadate at micromolar levels reversibly inhibits dynein ATPase activity and movement of demembranated flagella and cilia. It does not affect glycerinated myofibril contraction or myosin ATPase activty at less than millimolar concentrations. Vanadate at 10-- 100 micron reversibly inhibits anaphase movement of chromosomes and spindle elongation. After lysis in vanadate, spindles lose their fusiform appearance and become more barrel shaped. In vitro microtubule polymerization is insensitive to vanadate. PMID:152767

Arsenic removal in conjunction with lime softening

DOEpatents

Khandaker, Nadim R.; Brady, Patrick V.; Teter, David M.; Krumhansl, James L.

2004-10-12

A method for removing dissolved arsenic from an aqueous medium comprising adding lime to the aqueous medium, and adding one or more sources of divalent metal ions other than calcium and magnesium to the aqueous medium, whereby dissolved arsenic in the aqueous medium is reduced to a lower level than possible if only the step of adding lime were performed. Also a composition of matter for removing dissolved arsenic from an aqueous medium comprising lime and one or more sources of divalent copper and/or zinc metal ions.

Acoustic-wave-damping experimental study in x-ray preionized high-repetition rate XeCl

NASA Astrophysics Data System (ADS)

Canarelli, Patrick; Sentis, Marc L.; Delaporte, Philippe C.; Fontaine, Bernard L.; Forestier, Bernard M.

1990-08-01

To increase the pulse repetition frequency, the average power, and the beam quality of excimer laser systems, damping of the strong acoustic waves induced by the active medium excitation must be solve. In order to achieve this goal different electrodes and acoustic damping configuration have been studied. Excitation of active medium (energy deposition : 50 to 150 J/l) at high repetition rate (up to 1 000 Hz) in a subsonic loop (flow velocity : up to 65 m/s) is achieved by means of a classical discharge, through transfers capacitors. The discharge is preionized by X-Ray generated by a wire ion plasma gun. Previous studies done at I.M.F.M. have shown that the density perturbations, induced by successive excitations at high repetition rate (up to 1000 Hz), can involve output energy decrease. Here, we will present and discuss the density or pressure perturbation maximum level which don't involve this laser energy decrease; acoustic and thermal effects have been notably took account. We will study the possibilities of fast pressure perturbations damping.

Carbon nanotubes buckypaper radiation studies for medical physics applications.

PubMed

Alanazi, Abdulaziz; Alkhorayef, Mohammed; Alzimami, Khalid; Jurewicz, Izabela; Abuhadi, Nouf; Dalton, Alan; Bradley, D A

2016-11-01

Graphite ion chambers and semiconductor diode detectors have been used to make measurements in phantoms but these active devices represent a clear disadvantage when considered for in vivo dosimetry. In such circumstance, dosimeters with atomic number similar to human tissue are needed. Carbon nanotubes have properties that potentially meet the demand, requiring low voltage in active devices and an atomic number similar to adipose tissue. In this study, single-wall carbon nanotubes (SWCNTs) buckypaper has been used to measure the beta particle dose deposited from a strontium-90 source, the medium displaying thermoluminescence at potentially useful sensitivity. As an example, the samples show a clear response for a dose of 2Gy. This finding suggests that carbon nanotubes can be used as a passive dosimeter specifically for the high levels of radiation exposures used in radiation therapy. Furthermore, the finding points towards further potential applications such as for space radiation measurements, not least because the medium satisfies a demand for light but strong materials of minimal capacitance. Copyright © 2016 Elsevier Ltd. All rights reserved.

Pretreatment of Hanford medium-curie wastes by fractional crystallization.

PubMed

Nassif, Laurent; Dumont, George; Alysouri, Hatem; Rousseau, Ronald W

2008-07-01

Acceleration of the schedule for decontamination of the Hanford site using bulk vitrification requires implementation of a pretreatment operation. Medium-curie waste must be separated into two fractions: one is to go to a waste treatment and immobilization plant and a second, which is low-activity waste, is to be processed by bulk vitrification. The work described here reports research on using fractional crystallization for that pretreatment. Sodium salts are crystallized by evaporation of water from solutions simulating those removed from single-shell tanks, while leaving cesium in solution. The crystalline products are then recovered and qualified as low-activity waste, which is suitable upon redissolution for processing by bulk vitrification. The experimental program used semibatch operation in which a feed solution was continuously added to maintain a constant level in the crystallizer while evaporating water. The slurry recovered at the end of a run was filtered to recover product crystals, which were then analyzed to determine their composition. The results demonstrated that targets on cesium separation from the solids, fractional recovery of sodium salts, and sulfate content of the recovered salts can be achieved by the process tested.

[Protective effect of amlodipine on the cytotoxicity induced by contrast media in human kidney cells].

PubMed

Zhou, Xiao-rong; Duan, Shao-bin; Peng, You-ming; Liu, Fu-you; Ye, Yun; Liu, Rui-hong; Li, Gui-yuan

2007-10-01

To explore the protective effect of amlodipine on the cytotoxicity induced by contrast media (meglumine diatrizoate) in human kidney cells (HKC). An HKC line was used. The experiment was divided into 4 groups: a model group (diatrizoate 111g/L), a prevention group (diatrizoate 111g/L+amlodipine 10(-5)mol/L), an amlodipine control group (amlodipine 10(-5)mol/L), and a culture medium control group (simple none blood serum DMEM-F12 medium). Cytotoxicity induced by meglumine diatrizoate was analysed by methyl thiazolyl tetrazolium (MTT) test, lactate dehydrogenase (LDH) assay, Hochest33258 fluorescence stained cytospins, and flow cytometric DNA analysis. The protein expression of Bax was determined by Western blot, and caspase-3 activity was examined by fluorometric method. In the prevention group, the cell viability increased significantly (P<0.05), LDH levels decreased (P<0.05), and the apoptosis was lower than that of the model group (P<0.05) .Bax protein expression and caspase 3 activity decreased (P<0.05). Amlodipine can inhibit the HKC apoptosis and protect the renal tubule cell from injury induced by meglumine diatrizoate.

MANGO Imager Network Observations of Geomagnetic Storm Impact on Midlatitude 630 nm Airglow Emissions

NASA Astrophysics Data System (ADS)

Kendall, E. A.; Bhatt, A.

2017-12-01

The Midlatitude Allsky-imaging Network for GeoSpace Observations (MANGO) is a network of imagers filtered at 630 nm spread across the continental United States. MANGO is used to image large-scale airglow and aurora features and observes the generation, propagation, and dissipation of medium and large-scale wave activity in the subauroral, mid and low-latitude thermosphere. This network consists of seven all-sky imagers providing continuous coverage over the United States and extending south into Mexico. This network sees high levels of medium and large scale wave activity due to both neutral and geomagnetic storm forcing. The geomagnetic storm observations largely fall into two categories: Stable Auroral Red (SAR) arcs and Large-scale traveling ionospheric disturbances (LSTIDs). In addition, less-often observed effects include anomalous airglow brightening, bright swirls, and frozen-in traveling structures. We will present an analysis of multiple events observed over four years of MANGO network operation. We will provide both statistics on the cumulative observations and a case study of the "Memorial Day Storm" on May 27, 2017.

High Frontier, The Journal for Space & Missile Professionals. Volume 3, Number 4, August 2007

DTIC Science & Technology

2007-08-01

effective . Space power theories based on satellites and historical space activ - ity may be reasonably successful at adding structure to current...provider of combat effects for all mediums and levels of warfare should be proud of their accom- plishments. As we engage in global combat operations...we see everyday how our space and missile forces play a significant role in support of land, sea, and air combat operations. Space effects are

'Partners in clime': sustainable development and climate change--what can the National Health Service do?

PubMed

Cosford, P

2009-01-01

Climate change is arguably the biggest threat to health in the medium and long term. Necessary responses to this threat include adaptation, i.e. preparing to respond to the consequences of climate change, of which there are many in respect of health; and mitigation, i.e. reducing the activities that lead to climate change and, in particular, reducing the levels of greenhouse gas emissions, the most significant of which is carbon.

Caffeine, dibutyryl cyclic-AMP and heparin affect the chemotactic and phagocytotic activities of neutrophils for boar sperm in vitro.

PubMed

Li, J-C; Yamaguchi, S; Kondo, Y; Funahashi, H

2011-04-15

The objective was to examine the effects of caffeine, dibutyryl cyclic AMP, and heparin on the chemotaxis and/or phagocytosis of PMNs for porcine sperm. The chemotactic activity of PMNs, determined in a blind well chamber, increased (P < 0.05) when fresh serum was added to the medium (control containing BSA, 1109.5 cells/mm(2) vs serum, 1226.3 cells/mm(2)), regardless of the presence of sperm (control, 1121.1 cells/mm(2) vs serum, 1245.8 cells/mm(2)), whereas heat-inactivated serum did not affect activity (without sperm, 1099.4 cells/mm(2) and with sperm, 1132.6 cells/mm(2)). Regardless of live and dead sperm and of the origin of PMNs (boars vs sows), the phagocytotic activity of PMNs, as determined by co-culture of PMNs with sperm for 60 min, increased (P < 0.05) in the presence of fresh serum containing active complement (46.7 and 43.0%, respectively), but stimulation was decreased (P < 0.05) when 1 mM or higher concentrations of caffeine was added to the medium (from 40.7 to 20.8-31.6%). The origin of PMNs (sows vs boars) did not significantly affect phagocytotic activity. The percentage of PMNs that phagocytized polystyrene latex beads decreased when 2 mM caffeine was added to the medium containing porcine serum (from 43.7 to 21.5%). Serum-stimulated chemotactic activity of PMNs (1089.9 cells/mm(2)) was also reduced (P < 0.05) with 2 mM caffeine (942.5 cells/mm(2)). Furthermore, dibutyryl cAMP at ≥ 0.1 mM or heparin at ≥ 100 μg/mL decreased phagocytotic activity, in a concentration-dependent manner (P < 0.05). Supplementation of PMNs with heparin at 100 or 500 μg/mL decreased (P < 0.05) chemotactic activity in the presence of serum (from 1137.1 cells/mm(2) to 1008.8-1026.3 cells/mm(2)). We inferred that opsonization in the presence of active complement stimulated phagocytotic and chemotactic activities of PMNs, whereas supplementation with caffeine and dibutyryl cAMP (which could be associated with the intracellular cAMP level of PMNs) or adding heparin decreased serum-stimulated phagocytotic and chemotactic activities. Crown Copyright © 2011. Published by Elsevier Inc. All rights reserved.

Extraction, purification and anti-fatigue activity of γ-aminobutyric acid from mulberry (Morus alba L.) leaves

PubMed Central

Chen, Hengwen; He, Xuanhui; Liu, Yan; Li, Jun; He, Qingyong; Zhang, Cuiying; Wei, Benjun; Zhang, Ye; Wang, Jie

2016-01-01

Mulberry (Morus alba L.) is a tree species of Moraceae widely distributed in Southern China. In the present study, the white crystal of γ-aminobutyric acid (GABA) was purified from mulberry leaves, and its bioactivity was also investigated. The main results were as follows: first, the crude GABA was extracted from mulberry leaves by using biochemical methods. Then, the crude was purified by chromatography over an S-8 macroporous resin, Sephadex G-10, and 732 cation exchange resin to yield a white crystal. Lavage administration and exposure of GABA to male NIH mice showed no adverse effects on their growth and development. In an endurance capacity test, the average loaded-swimming time of medium dose was 111.60% longer than the control (P < 0.01). Further investigations showed that relative to that of model control, the respective blood lactate (BL) concentrations of low- and medium-dose were 28.52% and 28.81% lower (P < 0.05), whereas the levels of blood urea nitrogen (BUN) were 36.83% and 40.54% lower (P < 0.05), and that of liver glycogen (LG) levels were 12.81% and 17.22% lower (P < 0.05). The results indicated that GABA has an advantage over taurine of anti-fatigue effect. These findings were indicative of the anti-fatigue activity of GABA. PMID:26743028

Extraction, purification and anti-fatigue activity of γ-aminobutyric acid from mulberry (Morus alba L.) leaves.

PubMed

Chen, Hengwen; He, Xuanhui; Liu, Yan; Li, Jun; He, Qingyong; Zhang, Cuiying; Wei, Benjun; Zhang, Ye; Wang, Jie

2016-01-08

Mulberry (Morus alba L.) is a tree species of Moraceae widely distributed in Southern China. In the present study, the white crystal of γ-aminobutyric acid (GABA) was purified from mulberry leaves, and its bioactivity was also investigated. The main results were as follows: first, the crude GABA was extracted from mulberry leaves by using biochemical methods. Then, the crude was purified by chromatography over an S-8 macroporous resin, Sephadex G-10, and 732 cation exchange resin to yield a white crystal. Lavage administration and exposure of GABA to male NIH mice showed no adverse effects on their growth and development. In an endurance capacity test, the average loaded-swimming time of medium dose was 111.60% longer than the control (P < 0.01). Further investigations showed that relative to that of model control, the respective blood lactate (BL) concentrations of low- and medium-dose were 28.52% and 28.81% lower (P < 0.05), whereas the levels of blood urea nitrogen (BUN) were 36.83% and 40.54% lower (P < 0.05), and that of liver glycogen (LG) levels were 12.81% and 17.22% lower (P < 0.05). The results indicated that GABA has an advantage over taurine of anti-fatigue effect. These findings were indicative of the anti-fatigue activity of GABA.

Medium-/long-term effects of a specific exercise protocol combined with patient education on spine mobility, chronic fatigue, pain, aerobic fitness and level of disability in fibromyalgia.

PubMed

Giannotti, Erika; Koutsikos, Konstantinos; Pigatto, Maurizia; Rampudda, Maria Elisa; Doria, Andrea; Masiero, Stefano

2014-01-01

To propose a rehabilitation protocol able to produce immediate and long-term beneficial effects on level of disability and overall performance in ADLs. Forty-one FM patients were randomized to an exercise and educational-behavioral programme group (experimental group, EG = 21) or to a control group (CG = 20). Each subject was evaluated before, at the end (T1), and after 6 months (T6) from the conclusion of the rehabilitation treatment using the Fibromyalgia Impact Questionnaire (FIQ), the visual analogue scale (VAS), the Health Assessment Questionnaire (HAQ), the fatigue severity scale (FSS), the 6-minute walking test (6MWT), tender points count (TPC), and spinal active range of motion. The exercise protocol included 20 sessions consisting in self-awareness, stretching, strengthening, spine flexibility, and aerobic exercises, which patients were subsequently educated to perform at home. The two groups were comparable at baseline. At T1, the EG showed a positive trend in FIQ, VAS, HAQ, and FSS scales and significant improvement in 6MWT and in most spinal active range of motion measurements (P between 0.001 and 0.04). The positive results were maintained at the follow-up. The proposed programme was well tolerated and produced immediate and medium-term beneficial effects improving function and strain endurance. This trial is registered with DRKS00005071 on DRKS.

Extraction, purification and anti-fatigue activity of γ-aminobutyric acid from mulberry (Morus alba L.) leaves

NASA Astrophysics Data System (ADS)

Chen, Hengwen; He, Xuanhui; Liu, Yan; Li, Jun; He, Qingyong; Zhang, Cuiying; Wei, Benjun; Zhang, Ye; Wang, Jie

2016-01-01

Mulberry (Morus alba L.) is a tree species of Moraceae widely distributed in Southern China. In the present study, the white crystal of γ-aminobutyric acid (GABA) was purified from mulberry leaves, and its bioactivity was also investigated. The main results were as follows: first, the crude GABA was extracted from mulberry leaves by using biochemical methods. Then, the crude was purified by chromatography over an S-8 macroporous resin, Sephadex G-10, and 732 cation exchange resin to yield a white crystal. Lavage administration and exposure of GABA to male NIH mice showed no adverse effects on their growth and development. In an endurance capacity test, the average loaded-swimming time of medium dose was 111.60% longer than the control (P < 0.01). Further investigations showed that relative to that of model control, the respective blood lactate (BL) concentrations of low- and medium-dose were 28.52% and 28.81% lower (P < 0.05), whereas the levels of blood urea nitrogen (BUN) were 36.83% and 40.54% lower (P < 0.05), and that of liver glycogen (LG) levels were 12.81% and 17.22% lower (P < 0.05). The results indicated that GABA has an advantage over taurine of anti-fatigue effect. These findings were indicative of the anti-fatigue activity of GABA.

Immunostimulatory Activity of Opuntia ficus-indica var. Saboten Cladodes Fermented by Lactobacillus plantarum and Bacillus subtilis in RAW 264.7 Macrophages.

PubMed

Hwang, Joon-Ho; Lim, Sang-Bin

2017-02-01

To increase the functionality of Opuntia ficus-indica var. saboten cladodes, it was fermented by Lactobacillus plantarum and Bacillus subtilis. Eighty percent methanol extracts were investigated for their effects on nitric oxide (NO) production, cytokine secretion, nuclear factor-κB (NF-κB) activity, and mitogen-activated protein kinase (MAPK) phosphorylation in RAW 264.7 cells. Methanol extracts of L. plantarum culture medium (LPCME) and B. subtilis culture medium (BSCME) did not affect lipopolysaccharide (LPS)-induced NO production but, at 500 μg/mL, increased interferon (IFN)-γ-induced NO production by 55.2 and 66.5 μM, respectively, in RAW 264.7 cells. In RAW 264.7 cells not treated with LPS and IFN-γ, LPCME did not affect NO production, but BSCME increased NO production significantly in a dose-dependent manner. In addition, BSCME induced the expression of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in RAW 264.7 cells in a dose-dependent manner. BSCME at 500 μg/mL increased TNF-α and IL-1β mRNA levels by 83.8% and 82.2%, respectively. BSCME increased NF-κB-dependent luciferase activity in a dose-dependent manner; 500 μg/mL BSCME increased activity 9.1-fold compared with the control. BSCME induced the phosphorylation of p38, c-JUN NH 2 -terminal protein kinase (JNK), and extracellular signal-regulated kinase (ERK) in a dose-dependent manner, but did not affect total ERK levels. In conclusion, BSCME exerted immunostimulatory effects, which were mediated by MAPK phosphorylation and NF-κB activation, resulting in increased TNF-α and IL-1β gene expression in RAW 264.7 macrophages. Therefore, BSCM shows promise for use as an immunostimulatory therapeutic.

Bovine alpha-lactalbumin stimulates mucus metabolism in gastric mucosa.

PubMed

Ushida, Y; Shimokawa, Y; Toida, T; Matsui, H; Takase, M

2007-02-01

Bovine alpha-lactalbumin (alpha-LA), a major milk protein, exerts strong gastroprotective activity against rat experimental gastric ulcers induced by ethanol or stress. To elucidate the mechanisms underlying this activity, the influence of alpha-LA on gastric mucus metabolism was investigated in vitro and in vivo. For the in vitro study, RGM1 cells (a rat gastric epithelial cell line) were selected for observation of the direct activity of alpha-LA on gastric mucosal cells and cultured in the presence of either alpha-LA or ovalbumin (OVA), a reference protein showing no gastroprotective activity. Amounts of synthesized and secreted mucin, a major component of mucus, were determined using [3H]glucosamine as a tracer, and prostaglandin E2 (PGE2) levels in the culture medium were determined by RIA. For the in vivo study, the thickness of the mucus gel layer, a protective barrier for gastric mucosa, was evaluated histochemically in rat gastric mucosa. alpha-Lactalbumin (3 mg/mL) significantly stimulated mucin synthesis and secretion in RGM1 cells and also increased PGE2 levels in the culture medium. In contrast, OVA showed no enhancing effects under identical conditions. Neither indomethacin, a cyclo-oxygenase inhibitor, nor AH23848, a prostaglandin EP4 receptor antagonist, affected alpha-LA-induced enhancement of mucin synthesis and secretion. In vivo, oral administration of alpha-LA (300 mg/kg x 3 times/d x 7 d) increased the thickness of the mucus gel layer in rats. These results indicate that alpha-LA fortifies the mucus gel layer by stimulating mucin production and secretion in gastric mucus-producing cells, and that this enhancing effect is independent of endogenous PGE2. Comparison of the efficacy of alpha-LA with OVA suggests that the activities observed in RGM1 cells are closely related to the gastroprotective effects in rat gastric ulcer models. In conclusion, alpha-LA stimulates mucus metabolism, and this action may be responsible for its gastroprotective activity.

FIBER AND INTEGRATED OPTICS. OTHER TOPICS IN QUANTUM ELECTRONICS: Determination of the saturation parameter in a fast-flow CO2 active medium

NASA Astrophysics Data System (ADS)

Lebedev, V. F.; Lysikov, A. Yu; Ryazanova, A. V.; Shalygin, S. V.

1990-05-01

A diagnostic method was developed for determination of such an important characteristic of a CO2 active medium as the energy stored in the vibrational degrees of freedom of molecules, which can be determined directly from the experimentally measured powers of probe beams without the need for any additional information on the medium and without any numerical calculations. Measurements were carried out using a real fast-flow system, which satisfied the conditions of validity of the proposed diagnostic method, and the correctness of the approach was confirmed. The results obtained demonstrated the efficiency of the excitation of the medium when the pump conditions were varied.

Enhanced reaction kinetics in biological cells

NASA Astrophysics Data System (ADS)

Loverdo, C.; Bénichou, O.; Moreau, M.; Voituriez, R.

2008-02-01

The cell cytoskeleton is a striking example of an `active' medium driven out-of-equilibrium by ATP hydrolysis. Such activity has been shown to have a spectacular impact on the mechanical and rheological properties of the cellular medium, as well as on its transport properties: a generic tracer particle freely diffuses as in a standard equilibrium medium, but also intermittently binds with random interaction times to motor proteins, which perform active ballistic excursions along cytoskeletal filaments. Here, we propose an analytical model of transport-limited reactions in active media, and show quantitatively how active transport can enhance reactivity for large enough tracers such as vesicles. We derive analytically the average interaction time with motor proteins that optimizes the reaction rate, and reveal remarkable universal features of the optimal configuration. We discuss why active transport may be beneficial in various biological examples: cell cytoskeleton, membranes and lamellipodia, and tubular structures such as axons.

The relationship of exercise to anovulatory cycles in female athletes: hormonal and physical characteristics.

PubMed

Russell, J B; Mitchell, D; Musey, P I; Collins, D C

1984-04-01

The objective of this study was to examine the mechanisms by which physical activity affects the menstrual cycle. Women with high, medium, and low levels of physical activity were compared for menstrual function, physical characteristics, and urinary and serum levels of luteinizing hormone, follicle-stimulating hormone, prolactin, estradiol-17 beta, and 2-hydroxyestrone. None of the physical characteristics other than age and muscle area were significantly different in the three groups. The percentage of body fat did not appear to be a factor in the amenorrhea induced by strenuous exercise, as the percent of body fat in all three groups was less than 22%. The group of athletes under strenuous exercise which correlated with oligomenorrhea had decreased serum levels of luteinizing hormone, prolactin, and estradiol-17 beta but elevated levels of 2-hydroxyestrone. These data suggest that anovulatory cycles are correlated with the amount of exercise and increased levels of catechol estrogens. Catecholamines and beta-endorphin elevated by exercise may interact to suppress luteinizing hormone release at the hypothalamic pituitary axis.

Physico-chemical properties and cytotoxic potential of Cordyceps sinensis metabolites.

PubMed

Lee, Eun-Jeong; Jang, Ka-Hee; Im, Seon-Young; Lee, Yoon-Kyung; Farooq, Muhammad; Farhoudi, Rozbeh; Lee, Dong-Jin

2015-01-01

This study was conducted to estimate the antioxidant activities, biochemical properties and biological activities of one of the entomopathogenic fungi, Cordyceps sinensis. Analysis of fungal metabolites indicated that the most abundant free sugar was glucose; the highest component of organic acids was citric acid from 10-day culture medium and the glutamate was the predominant amino acid observed from 3-day culture medium. Maximum total polyphenols and flavonoids were detected in the 15-day culture medium. For cytotoxicity test, three cancer cell lines, HepG2 (liver), MCF-7 (breast) and A549 (lung) were used. The IC50 values of the highest toxicity of HepG2 cell lines were observed from 10-day cultured medium, whereas the highest toxicity of MCF-7 and A549 was observed on 5-day cultured medium. This is the first study reporting on the strong antioxidant and cytotoxic potential of C. sinensis. Culture medium of C. sinensis may thus be used as an effective antioxidant and anticancer treatment of natural origin.

Targeting the expression of glutathione- and sulfate-dependent detoxification enzymes in HepG2 cells by oxygen in minimal and amino acid enriched medium.

PubMed

Usarek, Ewa; Graboń, Wojciech; Kaźmierczak, Beata; Barańczyk-Kuźma, Anna

2016-02-01

Cancer cells exhibit specific metabolism allowing them to survive and proliferate in various oxygen conditions and nutrients' availability. Hepatocytes are highly active metabolically and thus very sensitive to hypoxia. The purpose of the study was to investigate the effect of oxygen on the expression of phase II detoxification enzymes in hepatocellular carcinoma cells (HepG2) cultured in minimal and rich media (with nonessential amino acids and GSH). The cells were cultured at 1% hypoxia, 10% tissue normoxia, and 21% atmospheric normoxia. The total cell count was determined by trypan blue exclusion dye and the expression on mRNA level by RT-PCR. The result indicated that the expression of glutathione-dependent enzymes (GSTA, M, P, and GPX2) was sensitive to oxygen and medium type. At 1% hypoxia the enzyme expression (with the exception of GSTA) was higher in minimal compared to rich medium, whereas at 10% normoxia it was higher in the rich medium. The expression was oxygen-dependent in both types of medium. Among phenol sulfotransferase SULT1A1 was not sensitive to studied factors, whereas the expression of SULT1A3 was depended on oxygen only in minimal medium. It can be concluded that in HepG2 cells, the detoxification by conjugation with glutathione and, to a lower extent with sulfate, may be affected by hypoxia and/or limited nutrients' availability. Besides, because the data obtained at 10% oxygen significantly differ from those at 21%, the comparative studies on hypoxia should be performed in relation to 10% but not 21% oxygen. Copyright © 2015 Elsevier Inc. All rights reserved.

Response learning stimulates dendritic spine growth on dorsal striatal medium spiny neurons.

PubMed

Briones, Brandy A; Tang, Vincent D; Haye, Amanda E; Gould, Elizabeth

2018-06-23

Increases in the number and/or the size of dendritic spines, sites of excitatory synapses, have been linked to different types of learning as well as synaptic plasticity in several brain regions, including the hippocampus, sensory cortex, motor cortex, and cerebellum. By contrast, a previous study reported that training on a maze task requiring the dorsal striatum has no effect on medium spiny neuron dendritic spines in this area. These findings might suggest brain region-specific differences in levels of plasticity as well as different cellular processes underlying different types of learning. No previous studies have investigated whether dendritic spine density changes may be localized to specific subpopulations of medium spiny neurons, nor have they examined dendritic spines in rats trained on a dorsolateral striatum-dependent maze task in comparison to rats exposed to the same type of maze in the absence of training. To address these questions further, we labeled medium spiny neurons with the lipophilic dye DiI and stained for the protein product of immediate early gene zif 268, an indirect marker of neuronal activation, in both trained and untrained groups. We found a small but significant increase in dendritic spine density on medium spiny neurons of the dorsolateral striatum after short-term intensive training, along with robust increases in the density of spines with mushroom morphology coincident with reductions in the density of spines with thin morphology. However, these results were not associated with zif 268 expression. Our findings suggest that short-term intensive training on a dorsolateral striatum-dependent maze task induces rapid increases in dendritic spine density and maturation on medium spiny neurons of the dorsolateral striatum, an effect which may contribute to early acquisition of the learned response in maze training. Copyright © 2018. Published by Elsevier Inc.

Garcina cambogia leaf and seawater for tannase production by marine Aspergillus awamori BTMFW032 under slurry state fermentation.

PubMed

Beena, S P; Basheer, Soorej M; Bhat, Sarita G; Chandrasekaran, M

2011-12-01

Garcinia gummi-gutta (syn. G. cambogia, G. quaesita), known to have medicinal properties, was evaluated as a substrate and inducer for tannase production by a marine Aspergillus awamori BTMFW032, under slurry state fermentation using Czapekdox-minimal medium and sea water as the cultivation medium. Among the various natural tannin substrates evaluated, Garcinia leaf supported maximal tannase production. The cultivation conditions and components of the cultivation medium were optimized employing response surface methodology. The experimental results were fitted to a second-order polynomial model at a 92.2% level of significance (p < 0.0001). The maximal tannase activity was obtained in a slurry state medium containing 26.6%, w/v, Garcinia leaf, supplemented with 0.1% tannic acid as inducer. The optimum values of pH, temperature and inoculum concentration obtained were 5.0, 40 degrees C and 3%, respectively. A Box-Behnken model study of the fermentation conditions was carried out, and the best production of tannase was registered at 40 degrees C without agitation. Optimization strategy employing response surface methodology led to nearly 3-fold increase in the enzyme production from 26.2 U/mL obtained in unoptimized medium to 75.2 Units/mL in Box Behnken design, within 18 h of fermentation. It was observed that sea water could support maximal tannase production by A. awamori compared with other media suggesting that the sea water salts could have played an inducer role in expression of tannase encoding genes. To the best of our knowledge, this is the first report on production of tannase, an industrially important enzyme, utilizing Garcinia leaf as substrate under slurry state fermentation by marine A. awamori and sea water as the cultivation medium.

Formation of short high-power laser radiation pulses in excimer mediums

NASA Astrophysics Data System (ADS)

Losev, V. F., Sr.; Ivanov, N. G.; Panchenko, Yu. N.

2007-06-01

Presently an excimer mediums continue are examined as one of variants for formation of powerful and over powerful pulses of laser radiation with duration from units of nanosecond up to tens femtosecond. The researches on such powerful installations as "NIKE" (USA) and << SUPER ASHURA >>, Japan) proceed in this direction. The main advantage of excimer mediums is the opportunity to work in a frequency mode, absence of restriction on the size of active area, high uniformity of a gas working medium, high efficiency (up to 10 %) and wide spectral range of laser radiation (KrF, XeCl ~ 2nm, XeF (C-A), Xe IICl ~ 50-100 nanometers). Research in area of high quality laser beams formation in excimer mediums and its amplification in high power amplifiers are carried out the long time in Institute of High Current Electronics SB RAS, Tomsk, Russia. The wide aperture XeCl laser system of MELS-4k is used for these investigations. Last time we take part in program on development of high power excimer laser system with a petawatt level of power. This system supposes the formation and amplification high quality laser beams with different pulse duration from units of nanosecond up to tens femtosecond. We research the possibility of laser beams formation in excimer mediums with ps-ns pulse duration having the low noise and divergence near to diffraction limit. In other hand, we are developing the wide aperture XeF(C-A) amplifier with optical pump on base electron accelerator. According to our estimations of the XeF(C-A) amplifier based on the converter of e-beam energy to the Xe II* fluorescence at 172 nm will allow to obtain up to 100 TW peak power in a 30 fs pulse.

Na/Ca exchange in the basolateral membrane of the A6 cell monolayer: role in Cai homeostasis.

PubMed

Brochiero, E; Raschi, C; Ehrenfeld, J

1995-05-01

The presence of a Na/Ca exchanger in A6 cells was investigated by measuring intracellular calcium (Cai) fluctuations and the 45Ca fluxes through the basolateral membranes (blm) of the cell monolayer. Removal of Na+ from the medium produced a transient increase in Cai followed by a regulatory phase returning Cai to control levels in 3-4 min, this phase being greatly accelerated (< 60 s) by NaCl addition (apparent Km of approximately 5 mM Na+). The Cai increase was only found with the Na(+)-free medium on the basolateral side of the cell monolayer. A twofold increase in the 45Ca influx was observed under these conditions. In Ca(2+)- depleted cells, the initial Cai increase after Ca2+ addition to the medium was greater when the putative Na/Ca exchanger was not functioning (i.e. in a Na(+)-free medium). 45Ca effluxes through the blm of the monolayer were greatly and transiently increased by a Na(+)-free medium on the serosal side and blocked by orthovanadate (1 mM). The Cai increased induced by a hypo-osmotic shock was greater in cells bathed in a Na(+)-medium, conditions expected to block the activity of the Na/Ca exchanger. These findings support the hypothesis that a Na/Ca exchanger is present on the blm of A6 cells and affirm its role in Cai homeostasis in steady-state conditions and following osmotic shock. In addition, a Ca2+ pump also located on the blm and Ca2+ stores sensitive to inositol 1,4,5-trisphosphate were found to be implicated in Cai homeostasis.

Cryptococcus Neoformans Modulates Extracellular Killing by Neutrophils

PubMed Central

Qureshi, Asfia; Grey, Angus; Rose, Kristie L.; Schey, Kevin L.; Del Poeta, Maurizio

2011-01-01

We recently established a key role for host sphingomyelin synthase (SMS) in regulating the killing activity of neutrophils against Cryptococcus neoformans. In this paper, we studied the effect of C. neoformans on the killing activity of neutrophils and whether SMS would still be a player against C. neoformans in immunocompromised mice lacking T and natural killer (NK) cells (Tgε26 mice). To this end, we analyzed whether C. neoformans would have any effect on neutrophil survival and killing in vitro and in vivo. We show that unlike Candida albicans, neither the presence nor the capsule size of C. neoformans cells have any effect on neutrophil viability. Interestingly, melanized C. neoformans cells totally abrogated the killing activity of neutrophils. We monitored how exposure of neutrophils to C. neoformans cells would interfere with any further killing activity of the conditioned medium and found that pre-incubation with live but not “heat-killed” fungal cells significantly inhibits further killing activity of the medium. We then studied whether activation of SMS at the site of C. neoformans infection is dependent on T and NK cells. Using matrix-assisted laser desorption–ionization tissue imaging in infected lung we found that similar to previous observations in the isogenic wild-type CBA/J mice, SM 16:0 levels are significantly elevated at the site of infection in mice lacking T and NK cells, but only at early time points. This study highlights that C. neoformans may negatively regulate the killing activity of neutrophils and that SMS activation in neutrophils appears to be partially independent of T and/or NK cells. PMID:21960987

Photonuclear activation of pure isotopic mediums.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Grohman, Mark A.; Lukosi, Eric Daniel

2010-06-01

This work simulated the response of idealized isotopic U-235, U-238, Th-232, and Pu-239 mediums to photonuclear activation with various photon energies. These simulations were conducted using MCNPX version 2.6.0. It was found that photon energies between 14-16 MeV produce the highest response with respect to neutron production rates from all photonuclear reactions. In all cases, Pu-239 responds the highest, followed by U-238. Th-232 produces more overall neutrons at lower photon energies then U-235 when material thickness is above 3.943 centimeters. The time it takes each isotopic material to reach stable neutron production rates in time is directly proportional to themore » material thickness and stopping power of the medium, where thicker mediums take longer to reach stable neutron production rates and thinner media display a neutron production plateau effect, due to the lack of significant attenuation of the activating photons in the isotopic mediums. At this time, no neutron sensor system has time resolutions capable of verifying these simulations, but various indirect methods are possible and should be explored for verification of these results.« less

Physical activity and associated socio-demographic factors in adolescents from the eastern region of Poland

PubMed

Wasilewska, Małgorzata; Bergier, Józef

The introduction and maintenance of healthy habits in the field of physical activity at the early stage of life is particularly important for public health. With increasing of public awareness in physical activity, researchers are increasingly questioning its determinants in different age groups. In this paper we focus on variables such as age, sex and place of residence. To determine what selected socio-demographic factors influence the level of physical activity of school youth from the eastern region of Poland in different domains of daily life. A random sample of 916 Polish school youth from Secondary Schools from Eastern Region of Poland aged 16-18 was interviewed in spring 2016 by diagnostic survey method with the use of the official Polish long version of the International Physical Activity Questionnaire (IPAQ). Analysing the relationship between gender and the level of physical activity statistically significant differences were found in all domains in favour of boys. The level of total physical activity in girls was 5345.5 MET-min./week, whereas in boys – 6556.6 MET-min./week. In the area of total physical activity, the highest values were observed in pupils from small towns and villages, with lower values in the youth living in large and medium-sized cities. When the relationship of age with the level of physical activity in particular domains was examined, only the activity in the work/ school domain (p = 0.0129) spoke in favour of the youngest pupils (2578.7 MET-min./week) when compared to the oldest ones (2226.4 MET-min./week). The level of physical activity in young people from the Eastern region of Poland is decreasing with age, both in girls and boys. It is therefore important, especially for the group emerging adulthood, to offer a wide variety of updated, involving, age and gender-oriented school physical activity classes that could be easily practised outside the school environment to improve the level of physical activity in leisure domain

Restrained management of copper level enhances the antineoplastic activity of imatinib in vitro and in vivo.

PubMed

Hassan, Iftekhar; Khan, Azmat Ali; Aman, Shazia; Qamar, Wajhul; Ebaid, Hossam; Al-Tamimi, Jameel; Alhazza, Ibrahim M; Rady, Ahmed M

2018-01-26

The present study was designed to investigate if elevated copper level can be targeted to enhance the efficacy of a significant anticancer drug, imatinib (ITB). The antineoplastic activity of this drug was assessed in the HepG2, HEK-293, MCF-7 and MDA-MD-231 cells targeting elevated copper level as their common drug target. The cell lines were treated with the different doses of copper chloride (Cu II) and disulfiram (DSF) alone as well as in their combinations with the drug for 24 h in standard culture medium and conditions. The treated cells were subjected to various assays including MTT, PARP, p-53, caspase-7, caspase-3, LDH and single cell electrophoresis. The study shows that DSF and Cu (II) synergizes the anticancer activity of ITB to a significant extent in a dose-specific way as evidenced by the combinations treated groups. Furthermore, the same treatment strategy was employed in cancer-induced rats in which the combinations of ITB-DSF and ITB-Cu II showed enhanced antineoplastic activity as compared to ITB alone. However, DSF was more effective than Cu (II) as an adjuvant to the drug. Hence, restrained manipulation of copper level in tumor cells can orchestrate the redox and molecular dispositions inside the cells favoring the induction of apoptosis.

Parental involvement and association with adolescents' fruit and vegetable intake at follow-up: Process evaluation results from the multi-component school-based Boost intervention.

PubMed

Jørgensen, Sanne Ellegård; Jørgensen, Thea Suldrup; Aarestrup, Anne Kristine; Due, Pernille; Krølner, Rikke

2016-10-26

Based on the assumption of parental influence on adolescent behavior, multicomponent school-based dietary interventions often include a parental component. The effect of this intervention component is seldom reported and the evidence is inconsistent. We conducted a systematic process evaluation of the parental component and examined whether the leveal of parental involvement in a large multi-component intervention: the Boost study was associated with adolescents' fruit and vegetable (FV) intake at follow-up. The Boost study was targeting FV intake among 1,175 Danish 7 th graders (≈13- year-olds) in the school year 2010/11. The study included a school component: free FV in class and curricular activities; a local community component: fact sheets for sports- and youth clubs; and a parental component: presentation of Boost at a parent-school meeting, 6 newsletters to parents, 3 guided student-parent curricular activities, and a student-parent Boost event. Students whose parent replied to the follow-up survey (n = 347). Questionnaire data from students, parents and teachers at 20 intervention schools. Process evaluation measures: dose delivered, dose received, appreciation and level of parental involvement. Parental involvement was trichotomized into: low/no (0-2 points), medium (3 points) and high (4-6 points). The association between level of parental involvement and self-reported FV intake (24-h recall), was analyzed using multilevel regression analyses. The Boost study was presented at a parent-school meeting at all intervention schools. The dose delivered was low to moderate for the three other parental elements. Most parents appreciated the intervention and talked with their child about Boost (83.5 %). High, medium and low parental involvement was found among 30.5 %, 29.6 % and 39.4 % of the students respectively. Parental involvement was highest among women. More men agreed that the parental newsletters provided new information. Students with a medium and high level of parental involvement ate 47.5 and 95.2 g more FV per day compared to students with low level/no parental involvement (p = 0.02). Students with a high level of parental involvement ate significantly more FV at follow-up compared to students with a low level/no parental involvement. Parental involvement in interventions may improve adolescents' FV intake if challenges of implementation can be overcome. ISRCTN11666034 . Registered 06/01/2012. Retrospectively registered.

The Investigation of ADAMTS16 in Insulin-Induced Human Chondrosarcoma Cells.

PubMed

Cakmak, Ozlem; Comertoglu, Ismail; Firat, Ridvan; Erdemli, Haci Kemal; Kursunlu, S Fatih; Akyol, Sumeyya; Ugurcu, Veli; Altuntas, Aynur; Adam, Bahattin; Demircan, Kadir

2015-08-01

A disintegrin-like metalloproteinase with thrombospondin motifs (ADAMTS) is a group of proteins that have enzymatic activity secreted by cells to the outside extracellular matrix. Insulin induces proteoglycan biosynthesis in chondrosarcoma chondrocytes. The purpose of the present in vitro study is to assess the time course effects of insulin on ADAMTS16 expression in OUMS-27 (human chondrosarcoma) cell line to examine whether insulin regulates ADAMTS16 expression as well as proteoglycan biosynthesis with multifaceted properties or not. Chondrosarcoma cells were cultured in Dulbecco's modified Eagle's medium having either 10 μg/mL insulin or not. While the experiment was going on, the medium containing insulin had been changed every other day. Cells were harvested at 1st, 3rd, 7th, and 11th days; subsequently, RNA and proteins were isolated in every experimental group according to their time interval. RNA expression of ADAMTS was estimated by quantitative real-time polymerase chain reaction (qRT-PCR) by using primers. Immunoreactive protein levels were encountered by the western blot protein detection technique by using proper anti-ADAMTS16 antibodies. ADAMTS16 mRNA expression level of chondrosarcoma cells was found to be insignificantly decreased in chondrosarcoma cells induced by insulin detected by the qRT-PCR instrument. On the other hand, there was a gradual decrease in immune-reactant ADAMTS16 protein amount by the time course in insulin-treated cell groups when compared with control cells. It has been suggested that insulin might possibly regulate ADAMTS16 levels/activities in OUMS-27 chondrosarcoma cells taking a role in extracellular matrix turnover.

Optical functional performance of the osteo-odonto-keratoprosthesis.

PubMed

Lee, Richard M H; Ong, Gek L; Lam, Fook Chang; White, Joy; Crook, David; Liu, Christopher S C; Hull, Chris C

2014-10-01

The aim of this study was to evaluate optical and visual functional performance of the osteo-odonto-keratoprosthesis (OOKP). Optical design and analysis was performed with customized optical design software. Nine patients with implanted OOKP devices and 9 age-matched control patients were assessed. Contrast sensitivity was assessed and glare effect was measured with a brightness acuity test. All OOKP patients underwent kinetic Goldmann perimetry and wavefront aberrometry and completed the National Eye Institute Visual Function Questionnaire-25 (NEI VFQ-25). Optical analysis showed that the optical cylinder is near diffraction-limited. A reduction in median visual acuity (VA) with increasing glare settings was observed from 0.04 logMAR (without glare) to 0.20 logMAR (with glare at "high" setting) and significantly reduced statistically when compared with the control group at all levels of glare (P < 0.05). Contrast sensitivity was significantly reduced when compared with age-matched controls at medium and high spatial frequencies (P < 0.05). Median Goldmann perimetry was 65 degrees (interquartile range, 64-74 degrees; V-4e isopters) and 69 degrees excluding 2 glaucomatous subjects. Several vision-related NEI VFQ-25 subscales correlated significantly with VA at various brightness acuity test levels and contrast sensitivity at medium spatial frequencies, including dependency, general vision, near activities and distance activities. The OOKP optical cylinder provides patients with a good level of VA that is significantly reduced by glare. We have shown in vivo that updates to the optical cylinder design have improved the patient's field of view. Reduction of glare and refinement of cylinder alignment methods may further improve visual function and patient satisfaction.

Regulation of Botulinum Neurotoxin Synthesis and Toxin Complex Formation by Arginine and Glucose in Clostridium botulinum ATCC 3502.

PubMed

Fredrick, Chase M; Lin, Guangyun; Johnson, Eric A

2017-07-01

Botulinum neurotoxin (BoNT), produced by neurotoxigenic clostridia, is the most potent biological toxin known and the causative agent of the paralytic disease botulism. The nutritional, environmental, and genetic regulation of BoNT synthesis, activation, stability, and toxin complex (TC) formation is not well studied. Previous studies indicated that growth and BoNT formation were affected by arginine and glucose in Clostridium botulinum types A and B. In the present study, C. botulinum ATCC 3502 was grown in toxin production medium (TPM) with different levels of arginine and glucose and of three products of arginine metabolism, citrulline, proline, and ornithine. Cultures were analyzed for growth (optical density at 600 nm [OD 600 ]), spore formation, and BoNT and TC formation by Western blotting and immunoprecipitation and for BoNT activity by mouse bioassay. A high level of arginine (20 g/liter) repressed BoNT production approximately 1,000-fold, enhanced growth, slowed lysis, and reduced endospore production by greater than 1,000-fold. Similar effects on toxin production were seen with equivalent levels of citrulline but not ornithine or proline. In TPM lacking glucose, levels of formation of BoNT/A1 and TC were significantly decreased, and extracellular BoNT and TC proteins were partially inactivated after the first day of culture. An understanding of the regulation of C. botulinum growth and BoNT and TC formation should be valuable in defining requirements for BoNT formation in foods and clinical samples, improving the quality of BoNT for pharmaceutical preparations, and elucidating the biological functions of BoNTs for the bacterium. IMPORTANCE Botulinum neurotoxin (BoNT) is a major food safety and bioterrorism concern and is also an important pharmaceutical, and yet the regulation of its synthesis, activation, and stability in culture media, foods, and clinical samples is not well understood. This paper provides insights into the effects of critical nutrients on growth, lysis, spore formation, BoNT and TC production, and stability of BoNTs of C. botulinum We show that for C. botulinum ATCC 3502 cultured in a complex medium, a high level of arginine repressed BoNT expression by ca. 1,000-fold and also strongly reduced sporulation. Arginine stimulated growth and compensated for a lack of glucose. BoNT and toxin complex proteins were partially inactivated in a complex medium lacking glucose. This work should aid in optimizing BoNT production for pharmaceutical uses, and furthermore, an understanding of the nutritional regulation of growth and BoNT formation may provide insights into growth and BoNT formation in foods and clinical samples and into the enigmatic function of BoNTs in nature. Copyright © 2017 American Society for Microbiology.

Plasminogen Activator Production Accompanies Loss of Anchorage Regulation in Transformation of Primary Rat Embryo Cells by Simian Virus 40

PubMed Central

Pollack, R.; Risser, R.; Conlon, S.; Rifkin, D.

1974-01-01

We have isolated several lines of rat embryo cells transformed by simian virus 40. All these lines are fully transformed with regard to saturation density and serum sensitivity, but they differ greatly in their anchorage dependence, as assayed by efficiency of plating in methyl cellulose suspension. This set of lines reveals a consistent relation of plasminogen activator production to plating efficiency in methyl cellulose. T-antigen-positive transformed lines that synthesize activator grow in methyl cellulose suspension, while T-antigen-positive transformed lines that do not synthesize activator fail to form colonies in suspension. Normal rat embryo cells produce very little plasminogen activator and do not grow in methyl cellulose. Sera that permit high levels of plasmin formation and activity support growth in semi-solid medium better than sera whose plasminogen is activated poorly and/or sera that contain inhibitors to plasmin. PMID:4373730

Quercetin improves the in vitro development of porcine oocytes by decreasing reactive oxygen species levels

PubMed Central

Kang, Jung-Taek; Kwon, Dae-Kee; Park, Sol-Ji; Kim, Su-Jin; Moon, Joon-Ho; Koo, Ok-Jae; Jang, Goo

2013-01-01

Quercetin is a plant-derived flavonoid found in fruits or vegetables that has antioxidant properties and acts as a free radical scavenger. We investigated the effects of quercetin on porcine oocyte nuclear maturation and embryonic development after parthenogenetic activation. We then evaluated the antioxidant activities of quercetin by measuring reactive oxygen species (ROS) levels in matured oocytes. Immature oocytes were untreated or treated with 1, 10, and 50 µg/mL quercetin during in vitro maturation (IVM). Quercetin treatment did not improve oocyte nuclear maturation, but significantly higher blastocyst rates (p < 0.05) of parthenogenetically activated oocytes were achieved when the IVM medium was supplemented with an adequate concentration of quercetin (1 µg/mL). However, cleavage rates and blastocyst cell numbers were not affected. Oocytes treated with 1 or 10 µg/mL quercetin had significantly lower (p < 0.05) levels of ROS than the control and group treated with the highest concentration of quercetin (50 µg/mL). Moreover, this highest concentration was detrimental to oocyte nuclear maturation and blastocyst formation. Based on our findings, we concluded that exogenous quercetin reduces ROS levels during oocyte maturation and is beneficial for subsequent embryo development. PMID:23388446

Using the Medium to Teach the Medium: Radio Broadcasting.

ERIC Educational Resources Information Center

Ryland, Edwin H.

Effective broadcast education should get students to perform professionally in broadcast situations with a minimum of guidance. To do this, class assignments should include either activities similar to commercial work or on-the-job experiences at the campus or local commercial radio stations. The possibilities for such activities and experiences…

Recovery of insulin sensitivity and optimal body composition after rapid weight loss in obese dogs fed a high-protein medium-carbohydrate diet.

PubMed

André, A; Leriche, I; Chaix, G; Thorin, C; Burger, M; Nguyen, P

2017-06-01

This study investigated the effects of an experimental high-protein medium-carbohydrate diet (protein level, 46% metabolizable energy, ME). First, postprandial plasma glucose and insulin kinetics were determined in steady-state overweight/obese Beagle dogs (28%-41% excess body weight) for an experimental high-protein medium-carbohydrate diet (protein level, 46% ME) and a commercial high-carbohydrate medium-protein diet (protein level, 24%ME) in obese dogs. Secondly, all the dogs were included in a weight loss programme. They were fed the high-protein medium-carbohydrate diet, and the energy allocation was gradually reduced until they reached their optimal body weight. Insulin sensitivity and body composition were evaluated before and after weight loss using a euglycaemic-hyperinsulinaemic clamp and the deuterium oxide dilution technique respectively. For statistical analysis, linear mixed effect models were used with a significance level of 5%. Postprandial plasma glucose and insulin concentrations were substantially lower with the high-protein medium-carbohydrate diet than the high-carbohydrate medium-protein diet. These differences can be explained mainly by the difference in carbohydrate content between the two diets. Energy restriction (35% lower energy intake than in the obese state) resulted in a 2.23 ± 0.05% loss in body weight/week, and the dogs reached their optimal body weight in 12-16 weeks. Weight loss was associated with a significant increase in insulin sensitivity. The high-protein medium-carbohydrate diet allowed fat-free mass preservation despite a relatively high rate of weekly weight loss. The increase in insulin sensitivity indicated improved control of carbohydrate metabolism, possible due to weight loss and to the nature of the diet. Thus, a high-protein medium-carbohydrate diet is a good nutritional solution for managing the weight of overweight dogs. This diet may improve glycaemic control, which could be beneficial for preventing or managing impaired glucose tolerance in obese dogs and for safe and successful weight loss while preserving lean body mass. Journal of Animal Physiology and Animal Nutrition © 2017 Blackwell Verlag GmbH.

20 CFR Appendix 2 to Part 220 - Medical-Vocational Guidelines

Code of Federal Regulations, 2012 CFR

2012-04-01

... medically determinable impairment(s). 203.00Maximum sustained work capability limited to medium work as a... his or her maximum sustained work capability for sedentary, light, medium, heavy, or very heavy work... at the various functional levels (sedentary, light, medium, heavy, and very heavy) as supported by...

20 CFR Appendix 2 to Part 220 - Medical-Vocational Guidelines

Code of Federal Regulations, 2013 CFR

2013-04-01

... medically determinable impairment(s). 203.00Maximum sustained work capability limited to medium work as a... his or her maximum sustained work capability for sedentary, light, medium, heavy, or very heavy work... at the various functional levels (sedentary, light, medium, heavy, and very heavy) as supported by...

20 CFR Appendix 2 to Part 220 - Medical-Vocational Guidelines

Code of Federal Regulations, 2014 CFR

2014-04-01

... medically determinable impairment(s). 203.00Maximum sustained work capability limited to medium work as a... his or her maximum sustained work capability for sedentary, light, medium, heavy, or very heavy work... at the various functional levels (sedentary, light, medium, heavy, and very heavy) as supported by...

20 CFR Appendix 2 to Part 220 - Medical-Vocational Guidelines

Code of Federal Regulations, 2011 CFR

2011-04-01

... medically determinable impairment(s). 203.00Maximum sustained work capability limited to medium work as a... his or her maximum sustained work capability for sedentary, light, medium, heavy, or very heavy work... at the various functional levels (sedentary, light, medium, heavy, and very heavy) as supported by...

20 CFR Appendix 2 to Part 220 - Medical-Vocational Guidelines

Code of Federal Regulations, 2010 CFR

2010-04-01

... medically determinable impairment(s). 203.00Maximum sustained work capability limited to medium work as a... his or her maximum sustained work capability for sedentary, light, medium, heavy, or very heavy work... at the various functional levels (sedentary, light, medium, heavy, and very heavy) as supported by...

Evaluation and analysis of noise levels at traffic intersections of Nagpur city, India.

PubMed

Vijay, Ritesh; Popat, Rishabh; Pisode, Mayur; Sharma, Asheesh; Manoj, Kumar; Chakrabarti, T; Gupta, Rajesh

2013-04-01

The objective of the present study was to monitor and assess the noise levels at traffic intersections in Napgur city under heterogenic traffic activities. For this, traffic volume and noise level were measured at intersections on highways, major roads andring road during morning and evening peak hours. Traffic volume was categorized in light, medium and heavy vehicles while noise levels were measured for Lmin, Lmax, Leq , L10 and L90. Equivalent noise was observed in the range of 71.3 to 79.3 dB(A) at the traffic intersections. Due to heterogenic traffic conditions and activities at the intersection like honking, idling, gear noise, bearing noise, breaking noise, tyre-road noise and exhaust noise, no correlation was established between traffic volume and observed noise levels except West High Court road. A strong correlation was found at West High Court road due to controlled traffic flow and less impact of heavy vehicles. Impact of noisy vehicles on general traffic was also assessed at the traffic intersections based on noise pollution levels and traffic noise index. The study suggests that control measures are required at the traffic intersections to minimize noise pollution levels.

Relating to ion detection

DOEpatents

Orr, Christopher Henry; Luff, Craig Janson; Dockray, Thomas; Macarthur, Duncan Whittemore

2001-01-01

The apparatus and method provide a technique for improving detection of alpha and/or beta emitting sources on items or in locations using indirect means. The emission forms generate ions in a medium surrounding the item or location and the medium is then moved to a detecting location where the ions are discharged to give a measure of the emission levels. To increase the level of ions generated and render the system particularly applicable for narrow pipes and other forms of conduits, the medium pressure is increased above atmospheric pressure. STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

THE LOCALIZATION OF MYOFIBRILLAR ATPASE ACTIVITY IN THE FLIGHT MUSCLES OF THE BLOWFLY, CALLIPHORA ERYTHROCEPHALA

PubMed Central

Tice, Lois W.; Smith, David S.

1965-01-01

The distribution of ATPase activity in the asynchronous flight muscles of Calliphora erythrocephala (Diptera) was studied at a fine structural level, using preparations of teased fibers, both unfixed and after brief fixation in hydroxyadipaldehyde, incubated in a medium for the histochemical demonstration of myosin or actomyosin ATPase. In relaxed fibrils, activity was found confined to the A bands and was absent from the H zones as well as from the Z and I band regions. At high magnification, deposits of final product, lead phosphate, appeared primarily related to the thick filaments, or to short lateral extensions from them. Evidence was gathered which indicated that this enzyme activity was that of a triphosphatase which did not act on dinucleoside or non-nucleoside substrates. PMID:4221034

[Sexuality in adolescents].

PubMed

Molina, R; Araa, S; Ibazeta, G; Jordan, P; Lagos, E

1987-01-01

A survey of knowledge, attitude, and practices regarding human reproduction and sexuality was undertaken in 2 groups of secondary school students in Chile to assess whether greater knowledge of reproduction and sexuality is associated with greater permissiveness and earlier initiation of sexual activity. Students in 2 public schools, 1 coeducational and 1 for female students only, were of lower middle class background, while students at the coeducational private school were of higher socioeconomic status. An anonymous, semiclosed questionnaire was administered to students in the 3 schools. The schools were selected because their directors agreed to permit the study. 14.8% of the 351 public school students were aged 14 or under and 77.8% were 15-18, while 99.5% of the 197 private school students were aged 15-18. The students' levels of knowledge of human reproduction and sexuality were measured through direct personal assessments by the students themselves and through 21 questions to confirm the assessments. At least 93% of students in all schools said their level of knowledge was medium or high, but the test indicated that only 64% of public school students and 75% of private school students actually had medium or high levels of knowledge. 45.9% of private and 27.9% of public school students felt the information they received from their schools about sexuality was adequate, while 41.9% of private and 60.9% of public school students felt it was insufficient. There were no significant differences in the opinions of the 2 groups of students concerning premarital sex, but the reasons given by the private school students to explain their attitudes expressed a greater sense of commitment to the partner, while those of the public school students tended to be more functional. Among public school students, 38.7% of males and 9.7% of females reported having had sexual relations, while among private school students, 17.7% of males and 4.4% of females reported having done so. No significant differences were reported in the age of initiating sexual activity in the 2 groups. Students in the school with the highest overall level of knowledge of human reproduction and sexuality had lower overall levels of sexual activity.

Cuticle-degrading proteases produced by the entomopathogenic fungus Beauveria bassiana in the presence of coffee berry borer cuticle

PubMed Central

Dias, B.A.; Neves, P.M.O.J.; Furlaneto-Maia, L.; Furlaneto, M.C.

2008-01-01

A Brazilian isolate of Beauveria bassiana (CG425) that shows high virulence against the coffee berry borer (CBB) was examined for the production of subtilisin-like (Pr1) and trypsin-like (Pr2) cuticle-degrading proteases. Fungal growth was either in nitrate-medium or in CBB cuticle-containing medium under both buffered and unbuffered conditions. In unbuffered medium supplemented with cuticle, the pH of cultures dropped and Pr1 and Pr2 activities were detected in high amounts only at a pH of 5.5 or higher. In buffered cultures, Pr1 and Pr2 activities were higher in medium supplemented with cuticle compared to activities with nitrate-medium. The Pr1 and Pr2 activities detected were mostly in the culture supernatant. These data suggest that Pr1 and Pr2 proteases produced by strain CG425 are induced by components of CBB cuticle, and that the culture pH influences the expression of these proteases, indicating the occurrence of an efficient mechanism of protein secretion in this fungus. The results obtained in this study extend the knowledge about protease production in B. bassiana CG425, opening new avenues for studying the role of secreted proteases in virulence against the coffee berry borer during the infection process. PMID:24031220

An HPLC method for the determination of selected amino acids in human embryo culture medium.

PubMed

Drábková, Petra; Andrlová, Lenka; Kanďár, Roman

2017-02-01

A method for the determination of selected amino acids in culture medium using HPLC with fluorescence detection is described. Twenty hours after intra-cytoplasmic sperm injection, one randomly selected zygote was transferred to the culture medium. After incubation (72 h after fertilization), the culture medium in which the embryo was incubated and blank medium was immediately stored at -80°C. Filtered medium samples were derivatized with ortho-phthalaldehyde (naphthalene-2,3-dicarboxaldehyde), forming highly fluorescent amino acids derivatives. Reverse-phase columns (LichroCART, Purospher STAR RP 18e or Ascentis Express C 18 ) were used for the separation. The derivatives were analyzed by gradient elution with a mobile phase containing ethanol and sodium dihydrogen phosphate. The analytical performance of this method is satisfactory for all amino acids; the intra-assay coefficients of variation were <10% and quantitative recoveries were between 95.5 and 104.4%. Changes in the levels of selected amino acids before and after human embryo cultivation were observed. After embryo incubation, the levels of all amino acids in the medium were increased, apart from aspartate and asparagine. After the cultivation of some embryos, amino acids which were not part of the medium were detected. Low amino acids turnover was observed in some embryos. Copyright © 2016 John Wiley & Sons, Ltd.

Protective effects of intermittent hydrostatic pressure on osteoarthritic chondrocytes activated by bacterial endotoxin in vitro.

PubMed

Lee, Mel S; Ikenoue, Takashi; Trindade, Michael C D; Wong, Neal; Goodman, Stuart B; Schurman, David J; Smith, R Lane

2003-01-01

The role of continuous passive motion (CPM) in the management of septic arthritis and inflammatory arthritis remains of interest. CPM produces cyclic variations in intraarticular pressure that facilitates transport of fluid, nutrients, and solutes within and/or across the joint and stimulates chondrocyte metabolism. However, the precise mechanisms mediating the responses of chondrocytes to joint motion remain unclear. This study tested the hypothesis that dynamic mechanical loading counteracts effects of bacterial lipopolysaccharide (LPS), an inflammatory mediator, on chondrocyte metabolism. Intermittent hydrostatic pressure (IHP) (10 MPa for 4 h) was applied to human chondrocytes pretreated with LPS (1 microg/ml for 18 h). LPS activation of chondrocytes decreased mRNA signal levels of type II collagen by 67% and aggrecan by 56% and increased nitric oxide by 3.1-fold, monocyte chemotactic protein-1 mRNA signal levels by 6.5-fold, and matrix metalloproteinase-2 mRNA signal levels by 1.3-fold. Application of IHP to LPS-activated chondrocytes decreased nitric oxide synthase mRNA signal levels and nitric oxide levels in the culture medium. Exposure of LPS-activated chondrocytes to IHP upregulated type II collagen and aggrecan mRNA signal levels by 1.7-fold, relative to chondrocytes activated by LPS and maintained without loading. In addition, application of IHP decreased the upregulation in signal levels of monocyte chemotactic factor-1 and matrix metalloproteinase-2 following LPS activation by 45% and 15%, respectively. These data show that mechanical loading counteract effects of inflammatory agents, such as bacterial LPS, and suggest that postinfection sequelae are influenced by the presence or absence of joint loading.

Differential regulation of Smad3 and of the type II transforming growth factor-β receptor in mitosis: implications for signaling.

PubMed

Hirschhorn, Tal; Barizilay, Lior; Smorodinsky, Nechama I; Ehrlich, Marcelo

2012-01-01

The response to transforming growth factor-β (TGF-β) depends on cellular context. This context is changed in mitosis through selective inhibition of vesicle trafficking, reduction in cell volume and the activation of mitotic kinases. We hypothesized that these alterations in cell context may induce a differential regulation of Smads and TGF-β receptors. We tested this hypothesis in mesenchymal-like ovarian cancer cells, arrested (or not) in mitosis with 2-methoxyestradiol (2ME2). In mitosis, without TGF-β stimulation, Smad3 was phosphorylated at the C-terminus and linker regions and localized to the mitotic spindle. Phosphorylated Smad3 interacted with the negative regulators of Smad signaling, Smurf2 and Ski, and failed to induce a transcriptional response. Moreover, in cells arrested in mitosis, Smad3 levels were progressively reduced. These phosphorylations and reduction in the levels of Smad3 depended on ERK activation and Mps1 kinase activity, and were abrogated by increasing the volume of cells arrested in mitosis with hypotonic medium. Furthermore, an Mps1-dependent phosphorylation of GFP-Smad3 was also observed upon its over-expression in interphase cells, suggesting a mechanism of negative regulation which counters increases in Smad3 concentration. Arrest in mitosis also induced a block in the clathrin-mediated endocytosis of the type II TGF-β receptor (TβRII). Moreover, following the stimulation of mitotic cells with TGF-β, the proteasome-mediated attenuation of TGF-β receptor activity, the degradation and clearance of TβRII from the plasma membrane, and the clearance of the TGF-β ligand from the medium were compromised, and the C-terminus phosphorylation of Smad3 was prolonged. We propose that the reduction in Smad3 levels, its linker phosphorylation, and its association with negative regulators (observed in mitosis prior to ligand stimulation) represent a signal attenuating mechanism. This mechanism is balanced by the retention of active TGF-β receptors at the plasma membrane. Together, both mechanisms allow for a regulated cellular response to TGF-β stimuli in mitosis.

Differential Regulation of Smad3 and of the Type II Transforming Growth Factor-β Receptor in Mitosis: Implications for Signaling

PubMed Central

Hirschhorn, Tal; Barizilay, Lior; Smorodinsky, Nechama I.; Ehrlich, Marcelo

2012-01-01

The response to transforming growth factor-β (TGF-β) depends on cellular context. This context is changed in mitosis through selective inhibition of vesicle trafficking, reduction in cell volume and the activation of mitotic kinases. We hypothesized that these alterations in cell context may induce a differential regulation of Smads and TGF-β receptors. We tested this hypothesis in mesenchymal-like ovarian cancer cells, arrested (or not) in mitosis with 2-methoxyestradiol (2ME2). In mitosis, without TGF-β stimulation, Smad3 was phosphorylated at the C-terminus and linker regions and localized to the mitotic spindle. Phosphorylated Smad3 interacted with the negative regulators of Smad signaling, Smurf2 and Ski, and failed to induce a transcriptional response. Moreover, in cells arrested in mitosis, Smad3 levels were progressively reduced. These phosphorylations and reduction in the levels of Smad3 depended on ERK activation and Mps1 kinase activity, and were abrogated by increasing the volume of cells arrested in mitosis with hypotonic medium. Furthermore, an Mps1-dependent phosphorylation of GFP-Smad3 was also observed upon its over-expression in interphase cells, suggesting a mechanism of negative regulation which counters increases in Smad3 concentration. Arrest in mitosis also induced a block in the clathrin-mediated endocytosis of the type II TGF-β receptor (TβRII). Moreover, following the stimulation of mitotic cells with TGF-β, the proteasome-mediated attenuation of TGF-β receptor activity, the degradation and clearance of TβRII from the plasma membrane, and the clearance of the TGF-β ligand from the medium were compromised, and the C-terminus phosphorylation of Smad3 was prolonged. We propose that the reduction in Smad3 levels, its linker phosphorylation, and its association with negative regulators (observed in mitosis prior to ligand stimulation) represent a signal attenuating mechanism. This mechanism is balanced by the retention of active TGF-β receptors at the plasma membrane. Together, both mechanisms allow for a regulated cellular response to TGF-β stimuli in mitosis. PMID:22927969

Role of L-lysine-alpha-ketoglutarate aminotransferase in catabolism of lysine as a nitrogen source for Rhodotorula glutinis.

PubMed Central

Kinzel, J J; Winston, M K; Bhattacharjee, J K

1983-01-01

Wild-type and saccharopine dehydrogenaseless mutant strains of Rhodotorula glutinis grew in minimal medium containing lysine as the sole nitrogen source and simultaneously accumulated, in the culture supernatant, large amounts of a product identified as alpha-aminoadipic-delta-semialdehyde. The saccharopine dehydrogenase and pipecolic acid oxidase levels remained unchanged in wild-type cells grown in the presence of ammonium or lysine as the nitrogen source. Lysine-alpha-ketoglutarate aminotransferase activity was demonstrated in ammonium-grown cells. This activity was depressed in cells grown in the presence of lysine as the sole source of nitrogen. PMID:6408065

The laser-diode-excited 5 d-4 f luminescence of Ce3+ and Pr3+ ions embedded into a BaR2F8 matrix

NASA Astrophysics Data System (ADS)

Pushkar', A. A.; Uvarova, T. V.; Kozlova, N. S.; Kuznetsov, S. Yu.; Uvarova, A. G.

2013-06-01

We show the possibility of obtaining UV luminescence from 5 d-4 f transitions of rare-earth ions in the BaY2F8: (Yb3+, Pr3+, Ce3+) crystal under upconversion excitation by standard laser diodes with lasing wavelengths of 960, 808, and 840 nm. Various upconversion mechanisms of pumping for populating the higher-lying energy levels of the active ions, as well as methods of adaptation of the active medium BaY2F8: (Yb3+, Pr3+, Ce3+) to these mechanisms, are considered.

Taurine Biosynthesis in a Fish Liver Cell Line (ZFL) Adapted to a Serum-Free Medium

PubMed Central

Liu, Chieh-Lun; Watson, Aaron M.; Place, Allen R.; Jagus, Rosemary

2017-01-01

Although taurine has been shown to play multiple important physiological roles in teleosts, little is known about the molecular mechanisms underlying dietary requirements. Cell lines can provide useful tools for deciphering biosynthetic pathways and their regulation. However, culture media and sera contain variable taurine levels. To provide a useful cell line for the investigation of taurine homeostasis, an adult zebrafish liver cell line (ZFL) has been adapted to a taurine-free medium by gradual accommodation to a commercially available synthetic medium, UltraMEM™-ITES. Here we show that ZFL cells are able to synthesize taurine and be maintained in medium without taurine. This has allowed for the investigation of the effects of taurine supplementation on cell growth, cellular amino acid pools, as well as the expression of the taurine biosynthetic pathway and taurine transporter genes in a defined fish cell type. After taurine supplementation, cellular taurine levels increase but hypotaurine levels stay constant, suggesting little suppression of taurine biosynthesis. Cellular methionine levels do not change after taurine addition, consistent with maintenance of taurine biosynthesis. The addition of taurine to cells grown in taurine-free medium has little effect on transcript levels of the biosynthetic pathway genes for cysteine dioxygenase (CDO), cysteine sulfinate decarboxylase (CSAD), or cysteamine dioxygenase (ADO). In contrast, supplementation with taurine causes a 30% reduction in transcript levels of the taurine transporter, TauT. This experimental approach can be tailored for the development of cell lines from aquaculture species for the elucidation of their taurine biosynthetic capacity. PMID:28587087

Effects of Growth Medium on Matrix-Assisted Laser Desorption–Ionization Time of Flight Mass Spectra: a Case Study of Acetic Acid Bacteria

PubMed Central

Wieme, Anneleen D.; Spitaels, Freek; Aerts, Maarten; De Bruyne, Katrien; Van Landschoot, Anita

2014-01-01

The effect of the growth medium used on the matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectra generated and its consequences for species and strain level differentiation of acetic acid bacteria (AAB) were determined by using a set of 25 strains. The strains were grown on five different culture media that yielded a total of more than 600 mass spectra, including technical and biological replicates. The results demonstrate that the culture medium can have a profound effect on the mass spectra of AAB as observed in the presence and varying signal intensities of peak classes, in particular when culture media do not sustain optimal growth. The observed growth medium effects do not disturb species level differentiation but strongly affect the potential for strain level differentiation. The data prove that a well-constructed and robust MALDI-TOF mass spectrometry identification database should comprise mass spectra of multiple reference strains per species grown on different culture media to facilitate species and strain level differentiation. PMID:24362425

[Stimulation of human hepatic stellate cells by cytochrome P4502E1-mediated oxidative stress].

PubMed

Li, Jing; Liu, Tian-hui; You, Hong; Xu, You-qing; Wang, Chen

2010-08-01

To explore the stimulation of human hepatic stellate cells by Cytochrome P4502E1-mediated oxidative stress. HepG2-line was transfected with human CYP2E1 plasmid (HepG2/CYP2E1) and empty plasmid (HepG2/PCI) respectively. The CYP2E1 expression was evaluated with RT-PCR and Western blot. MDA was measured in culture medium of HepG2 cell lines. LX2 was co-incubated with HepG2/CYP2E1, HepG2/PCI and HepG2 respectively. The level of hydroxyproline in culture medium was examined in 48 hours and the cells were lysated and total RNA and protein were extracted. COL-1 and MMP2 mRNA levels were detected by RT-PCR and analyzed semi-quantitatively. PICP proteins were measured by ELISA. Zymography was performed to investigate MMP2 enzymatic activities. (1) MDA from the HepG2 which (HepG2/CYP2E1)express human CYP2E1 (6.51+/-0.25) was significantly higher than that from the HepG2 which do not (HepG2/PCI) express human CYP2E1 (3.07+/-0.29) and HepG2 alone (2.57+/-0.29). (F=22.66, all P<0.01). (2) After co-incubated for 48 hours,the level of hydroxyproline in culture medium (35.24+/-3.52) excreted from CYP2E1/LX2 could significantly increase (F=58.89, P is less than 0.01). PICP protein (540.01+/-11.38) excreted from CYP2E1/LX2 was significantly increased (F=124.97, P<0.01). Zymography showed MMP2 gene expression and enzymatic activities of MMP2 had no difference among the groups (F=0.29, P>0.05) (F=0.33, P>0.05). CYP2E1 derived oxidative stress mediated stimulation of collagen I synthesis by hepatic stellate cells. Hydroxyproline excreted by LX2 was increased by CYP2E1. COL-1mRNA had no difference among the groups (F=0.73, P>0.05).

Screening of agricultural wastes as a medium production of catalase for enzymatic fuel cell by Neurospora crassa InaCC F226

NASA Astrophysics Data System (ADS)

Santoso, Pugoh; Yopi

2017-12-01

Explorations of local microorganisms from Indonesia that can produce of catalase are still limited. Neurospora crassa is a fungus which resulting of two kinds of catalase, namely catalase-1 and catalase-3. We studied the production of catalase by Neurospora crassa (no. F226) from Indonesia Culture Collection (InaCC) in Solid State Fermentation (SSF). Among four screened agro wastes (corn cob, rice straw, oil palm empty fruit bunches, and bagasse), rice straw and oil palm empty fruit bunches (OPEFB) were remarked as the most promising substrate suited for the excellent growth and adequate production of catalase. Based on the result, the method of solid state fermentation was suitable to production of catalase. It is caused that the medium served to maintain microbial growth and metabolism. The filamentous filament is more suitable for living on solid media because it has a high tolerance to low water activity, and it has a high potential to excrete hydrolytic enzymes that caused of its morphology. The filamentous filament morphology allows the fungus to form colonies and penetrate the solid substrates in order to obtain nutrients. The results showed that the highest catalase activity was obtained on rice straw and oil palm empty fruit bunches medium with catalase activity of 39.1 U/mL and 37,7 U/mL in 50% moisture content medium, respectively. Optimization of humidity and pH medium in the rice straw were investigated which is the highest activity obtained in 30% moisture content and pH medium of 6. The catalase activity was reached in the value of 53.761 U/mL and 56.903 U/mL by incubated 48 hours and 96 hours, respectively.

The influence of spent household batteries to the organic fraction of municipal solid wastes during composting.

PubMed

Komilis, Dimitrios; Bandi, Dimitra; Kakaronis, Georgios; Zouppouris, Georgios

2011-06-01

The objective of this work was to investigate the potential transfer of 9 heavy metals from spent household batteries (zinc-carbon and alkaline-manganese batteries) to the organic fraction of municipal solid wastes during active composting. Six runs were performed including one control and 2 replications. Eleven types of alkaline and non-alkaline batteries were added at 3 different levels to the organic fraction of municipal solid wastes, namely at percentages equal to 0.98% w/w (low), 5.2% w/w (medium) and 10.6% w/w (high). Experiments were performed in 230 l insulated plastic aerobic bioreactors under a dynamic air flow regime for up to 60 days. Iron, copper and nickel masses contained in the organic fraction of the wastes were found significantly higher in the high level runs compared to the corresponding masses in the control. No metal transfer was obtained in the low and medium level runs. Metal mass balance closures ranged from 51% to 176%. Metals' concentrations in the leachates were below 10 mg l⁻¹ for most metals, except iron, while an increasing concentration trend versus time was measured in the leachates of the high level runs. In all cases, the contents of 5 regulated heavy metals in all end products were below the Hellenic limits. Copyright © 2011 Elsevier B.V. All rights reserved.

Production of bacterial cellulose and enzyme from waste fiber sludge

PubMed Central

2013-01-01

Background Bacterial cellulose (BC) is a highly crystalline and mechanically stable nanopolymer, which has excellent potential as a material in many novel applications, especially if it can be produced in large amounts from an inexpensive feedstock. Waste fiber sludge, a residue with little or no value, originates from pulp mills and lignocellulosic biorefineries. A high cellulose and low lignin content contributes to making the fiber sludge suitable for bioconversion, even without a thermochemical pretreatment step. In this study, the possibility to combine production of BC and hydrolytic enzymes from fiber sludge was investigated. The BC was characterized using field-emission scanning electron microscopy and X-ray diffraction analysis, and its mechanical properties were investigated. Results Bacterial cellulose and enzymes were produced through sequential fermentations with the bacterium Gluconacetobacter xylinus and the filamentous fungus Trichoderma reesei. Fiber sludges from sulfate (SAFS) and sulfite (SIFS) processes were hydrolyzed enzymatically without prior thermochemical pretreatment and the resulting hydrolysates were used for BC production. The highest volumetric yields of BC from SAFS and SIFS were 11 and 10 g/L (DW), respectively. The BC yield on initial sugar in hydrolysate-based medium reached 0.3 g/g after seven days of cultivation. The tensile strength of wet BC from hydrolysate medium was about 0.04 MPa compared to about 0.03 MPa for BC from a glucose-based reference medium, while the crystallinity was slightly lower for BC from hydrolysate cultures. The spent hydrolysates were used for production of cellulase with T. reesei. The cellulase activity (CMCase activity) in spent SAFS and SIFS hydrolysates reached 5.2 U/mL (87 nkat/mL), which was similar to the activity level obtained in a reference medium containing equal amounts of reducing sugar. Conclusions It was shown that waste fiber sludge is a suitable raw material for production of bacterial cellulose and enzymes through sequential fermentation. The concept studied offers efficient utilization of the various components in fiber sludge hydrolysates and affords a possibility to combine production of two high value-added products using residual streams from pulp mills and biorefineries. Cellulase produced in this manner could tentatively be used to hydrolyze fresh fiber sludge to obtain medium suitable for production of BC in the same biorefinery. PMID:23414733

Effect of salinity medium on antioxidant and antidiabetic activity marine endophytic fungus of asperegillus elegans ptf 9

NASA Astrophysics Data System (ADS)

Mulyani, Hani; Artanti, Nina; Fitria, Irni; Filailla, Euis; Kandace, Yoice Sri; Udin, Linar Zalinar

2017-11-01

Our previous studies on screening of antioxidant activities from various endophytic fungi isolated from marine bioata showed that A. elegans PTF9 isolated from sea weed is one of the fungus that has good antioxidant activity. In current study we reported the effect of medium salinity (0, 3 and 10% salt in PDB medium) on antioxidant and antidiabetes activity of mycelium and filtrate ethyl acetate extracts of A. elegans Ptf 9. The antioxidant assay was conducted using DPPH free radical scavenging activity method. The antidiabetes assay was conducted using a-glucosidase inhibitory activity method. The results showed that the best antioxidant activity was obtained from filtrate extract of fungus cultures with 0% salt (IC50=1.56 ppm), whereas the best antidiabetes activity was obtained from filtrate extract of fungus culture with 10% salt (IC50= 3.64 ppm). Addition of salt reduced the antioxidant activity, but not the antidiabetes activity. The results suggest that A. elegans PTF9 showed potential for further studies on isolation of antioxidant and antidiabetes lead compounds that could be use for further development of new drugs.

Surveying hospital network structure in New York State: how are they structured?

PubMed

Nauenberg, E; Brewer, C S

2000-01-01

We determine the most common network structures in New York state. The taxonomy employed uses three structural dimensions: integration, complexity, and risk-sharing between organizations. Based on a survey conducted in 1996, the most common type of network (26.4 percent) had medium levels of integration, medium or high levels of complexity, and some risk-sharing. Also common were networks with low levels of integration, low levels of complexity, and no risk-sharing (22.1 percent).

Mechanisms for cellular NO oxidation and nitrite formation in lung epithelial cells.

PubMed

Zhao, Xue-Jun; Wang, Ling; Shiva, Sruti; Tejero, Jesus; Myerburg, Mike M; Wang, Jun; Frizzell, Sam; Gladwin, Mark T

2013-08-01

Airway lining fluid contains relatively high concentrations of nitrite, and arterial blood levels of nitrite are higher than venous levels, suggesting the lung epithelium may represent an important source of nitrite in vivo. To investigate whether lung epithelial cells possess the ability to convert NO to nitrite by oxidation, and the effect of oxygen reactions on nitrite formation, the NO donor DETA NONOate was incubated with or without A549 cells or primary human bronchial epithelial (HBE) cells for 24 h under normoxic (21% O2) and hypoxic (1% O2) conditions. Nitrite production was significantly increased under all conditions in the presence of A549 or HBE cells, suggesting that both A549 and HBE cells have the capacity to oxidize NO to nitrite even under low-oxygen conditions. The addition of oxyhemoglobin to the A549 cell medium decreased the production of nitrite, consistent with NO scavenging limiting nitrite formation. Heat-denatured A549 cells produced much lower nitrite and nitrate, suggesting an enzymatic activity is required. This NO oxidation activity was highest in membrane-bound proteins with molecular size <100kDa. In addition, 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one and cyanide inhibited formation of nitrite in A549 cells. It has been shown that ceruloplasmin (Cp) possesses an NO oxidase and nitrite synthase activity in plasma based on NO oxidation to nitrosonium cation. We observed that Cp is expressed intracellularly in lung epithelial A549 cells and secreted into the medium under basal conditions and during cytokine stimulation. However, an analysis of Cp expression level and activity measured via p-phenylenediamine oxidase activity assay revealed very low activity compared with plasma, suggesting that there is insufficient Cp to contribute to detectable NO oxidation to nitrite in A549 cells. Additionally, Cp levels were knocked down using siRNA by more than 75% in A549 cells, with no significant change in either nitrite or cellular S-nitrosothiol formation compared to scrambled siRNA control under basal conditions or cytokine stimulation. These data suggest that lung epithelial cells possess NO oxidase activity, which is enhanced in cell-membrane-associated proteins and not regulated by intracellular or secreted Cp, indicating that alternative NO oxidases determine hypoxic and normoxic nitrite formation from NO in human lung epithelial cells. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

Managers' Understanding of Workplace Health Promotion within Small and Medium-Sized Enterprises: A Phenomenological Study

ERIC Educational Resources Information Center

Moore, Ann; Parahoo, Kader; Fleming, Paul

2011-01-01

Objective: This study aimed at exploring managers' understanding of workplace health promotion and experiences of workplace health promotion activity within small and medium-sized enterprises. Design: A Heideggerian interpretive phenomenological methodology was adopted. Setting: This study was undertaken with small and medium-sized enterprise…

Pigeon pea waste as a novel, inexpensive, substrate for production of a thermostable alkaline protease from thermoalkalophilic Bacillus sp. JB-99.

PubMed

Johnvesly, B; Manjunath, B R; Naik, G R

2002-03-01

Thermoalkaliphilic Bacillus sp. JB-99 was grown in a 250 ml Erlenmeyer flask containing 50 ml medium containing (g/l) Pigeon pea waste 10; NaNO3, 5.0; K2HPO4, 5.0; MgSO4 x 2H2O, 0.2 and Na2CO3, 10.0. Incubations were carried out at 50 degrees C on a rotary incubator shaker for 15 h. A high level of extra cellular thermostable protease activity was observed after 24 h incubation. The optimum temperature and pH for activity were 70 degrees C and 11, respectively, so this enzyme showed stable activity at high temperature and under alkaline conditions.

Active microwave remote sensing of an anisotropic random medium layer

NASA Technical Reports Server (NTRS)

Lee, J. K.; Kong, J. A.

1985-01-01

A two-layer anisotropic random medium model has been developed to study the active remote sensing of the earth. The dyadic Green's function for a two-layer anisotropic medium is developed and used in conjunction with the first-order Born approximation to calculate the backscattering coefficients. It is shown that strong cross-polarization occurs in the single scattering process and is indispensable in the interpretation of radar measurements of sea ice at different frequencies, polarizations, and viewing angles. The effects of anisotropy on the angular responses of backscattering coefficients are also illustrated.

Fabrication update on critical-angle transmission gratings for soft x-ray grating spectrometers

NASA Astrophysics Data System (ADS)

Heilmann, Ralf K.; Bruccoleri, Alex; Mukherjee, Pran; Yam, Jonathan; Schattenburg, Mark L.

2011-09-01

Diffraction grating-based, wavelength dispersive high-resolution soft x-ray spectroscopy of celestial sources promises to reveal crucial data for the study of the Warm-Hot Intergalactic Medium, the Interstellar Medium, warm absorption and outflows in Active Galactic Nuclei, coronal emission from stars, and other areas of interest to the astrophysics community. Our recently developed critical-angle transmission (CAT) gratings combine the advantages of the Chandra high and medium energy transmission gratings (low mass, high tolerance of misalignments and figure errors, polarization insensitivity) with those of blazed reflection gratings (high broad band diffraction efficiency, high resolution through use of higher diffraction orders) such as the ones on XMM-Newton. Extensive instrument and system configuration studies have shown that a CAT grating-based spectrometer is an outstanding instrument capable of delivering resolving power on the order of 5,000 and high effective area, even with a telescope point-spread function on the order of many arc-seconds. We have fabricated freestanding, ultra-high aspect-ratio CAT grating bars from silicon-on-insulator wafers using both wet and dry etch processes. The 200 nm-period grating bars are supported by an integrated Level 1 support mesh, and a coarser external Level 2 support mesh. The resulting grating membrane is mounted to a frame, resulting in a grating facet. Many such facets comprise a grating array that provides light-weight coverage of large-area telescope apertures. Here we present fabrication results on the integration of CAT gratings and the different high-throughput support mesh levels and on membrane-frame bonding. We also summarize recent x-ray data analysis of 3 and 6 micron deep wet-etched CAT grating prototypes.

Timescale of silver nanoparticle transformation in neural cell cultures impacts measured cell response

NASA Astrophysics Data System (ADS)

Hume, Stephanie L.; Chiaramonti, Ann N.; Rice, Katherine P.; Schwindt, Rani K.; MacCuspie, Robert I.; Jeerage, Kavita M.

2015-07-01

Both serum protein concentration and ionic strength are important factors in nanoparticle transformation within cell culture environments. However, silver nanoparticles are not routinely tracked at their working concentration in the specific medium used for in vitro toxicology studies. Here we evaluated the transformation of electrostatically stabilized citrate nanoparticles (C-AgNPs) and sterically stabilized polyvinylpyrrolidone nanoparticles (PVP-AgNPs) in a low-serum ( 0.2 mg/mL bovine serum albumin) culture medium, while measuring the response of rat cortex neural progenitor cells, which differentiate in this culture environment. After 24 h, silver nanoparticles at concentrations up to 10 µg/mL did not affect adenosine triphosphate levels, whereas silver ions decreased adenosine triphosphate levels at concentrations of 1.1 µg/mL or higher. After 240 h, both silver nanoparticles, as well as silver ion, unambiguously decreased adenosine triphosphate levels at concentrations of 1 and 1.1 µg/mL, respectively, suggesting particle dissolution. Particle transformation was investigated in 1:10 diluted, 1:2 diluted, or undiluted differentiation medium, all having an identical protein concentration, to separate the effect of serum protein stabilization from ionic strength destabilization. Transmission electron microscopy images indicated that particles in 1:10 medium were not surrounded by proteins, whereas particles became clustered within a non-crystalline protein matrix after 24 h in 1:2 medium and at 0 h in undiluted medium. Despite evidence for a protein corona, particles were rapidly destabilized by high ionic strength media. Polyvinylpyrrolidone increased the stability of singly dispersed particles compared to citrate ligands; however, differences were negligible after 4 h in 1:2 medium or after 1 h in undiluted medium. Thus low-serum culture environments do not provide sufficient colloidal stability for long-term toxicology studies with citrate- or polyvinylpyrrolidone-stabilized silver nanoparticles.