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Constant Level (constant + level)
Selected AbstractsIdentification and characterization of Xenopus OMP25DEVELOPMENT GROWTH & DIFFERENTIATION, Issue 5 2004Masafumi Inui This study describes the isolation of mitochondrial outer membrane protein 25 (OMP25) from Xenopus laevis and an analysis of its role in early development. X. laevis OMP25 (xOMP25) is a transmembrane protein of the mitochondrial outer membrane with a PDZ domain in the cytoplasmic tail, and an approximate molecular size of 25 kDa. We isolated xOMP25 from a cDNA library of X. laevis tailbud embryos. Amino acid sequence analysis of xOMP25 showed 57% identity to mouse OMP25, with 73% identity in the PDZ domains. XOMP25 mRNA is expressed maternally, and at a constant level throughout early development. The transcript is localized to eye, otic vesicle, branchial arch and neural tube. Mitochondrial targeting of an EGFP-fusion protein of xOMP25 was visualized using a mitochondria-specific fluorescent dye. Overexpression of xOMP25 in embryos caused curved axes, small eyes and disorganized head structures. Knockdown of xOMP25 protein using antisense morpholino oligonucleotides resulted in slightly shortened axes and decreased neural tissue. Although the mechanism remains unclear, our results implicate xOMP25 protein in the formation of the intact neural tube. [source] Cloning and characterization of cDNA for syndecan core protein in sea urchin embryosDEVELOPMENT GROWTH & DIFFERENTIATION, Issue 5 2000Kazuo Tomita The cDNA for the core protein of the heparan sulfate proteoglycan, syndecan, of embryos of the sea urchin Anthocidaris crassispina was cloned and characterized. Reverse transcription,polymerase chain reaction (RT-PCR) was used with total ribonucleic acid (RNA) from late gastrula stage embryos and degenerate primers for conserved regions of the core protein, to obtain a 0.1 kb PCR product. A late gastrula stage cDNA library was then screened using the PCR product as a probe. The clones obtained contained an open reading frame of 219 amino acid residues. The predicted product was 41.6% identical to mouse syndecan-1 in the region spanning the cytoplasmic and transmembrane domains. Northern analysis showed that the transcripts were present in unfertilized eggs and maximum expression was detected at the early gastrula stage. Syndecan mRNA was localized around the nuclei at the early cleavage stage, but was then found in the ectodermal cells of the gastrula embryos. Western blotting analysis using the antibody against the recombinant syndecan showed that the proteoglycan was present at a constant level from the unfertilized egg stage through to the pluteus larval stage. Immunostaining revealed that the protein was expressed on apical and basal surfaces of the epithelial wall in blastulae and gastrulae. [source] Towards a New Articulation of Alternative Development: Lessons from Coca Supply Reduction in BoliviaDEVELOPMENT POLICY REVIEW, Issue 4 2004Noam Lupu Once heralded as the success story of coca supply reduction, Bolivia is now witnessing an increase in coca cultivation. Even as coca fields in Bolivia were forcibly destroyed in the past decade, new fields were being planted elsewhere, leaving coca production in the Andean region at a roughly constant level. This begs a rethinking of alternative development programmes, the policies being rendered ineffectual by the increasing use of force. This article seeks renewed momentum for alternative development by gleaning lessons from its earlier failures. Moreover, it suggests a new articulation of alternative development that emphasises the socio-economic cause of coca cultivation , the demand by the rural poor of Bolivia for income and food security. [source] Postprandial lipemic response to alpha-linolenic acid rich oil, butter, and olive oilEUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY, Issue 9 2010Julia Svensson Abstract Postprandial lipemia varies with composition of dietary fat due to partitioning of fatty acids between ,-oxidation, incorporation into TAG, and tissue lipids. Effects of alpha-linolenic acid (ALA) are poorly characterized. Lipase-catalyzed transesterification was used to produce a novel ALA-oil (35% ALA) from rapeseed and linseed oil. We hypothesized a lower postprandial lipemic response with ALA-oil than with olive oil and butter due to higher ,-oxidation of ALA. A randomized crossover study with 26 healthy men compared the effects on plasma lipids 7,h after a breakfast containing 35,g ALA-rich oil, butter fat, or olive oil. The incremental area under curve for plasma TAG was lower with butter than with olive oil (34%, p<0.05) and ALA-oil (25%, ns). After ALA-oil percentage ALA increased, in TAG to a constant level of 7,mol% and in NEFA to 6% after 7,h. Since total NEFA increased with time the amount of exogenous ALA in NEFA also increased. Butter resulted in lower postprandial lipemia than the oils, the difference exceeding what is expected from the presence of short and medium chain fatty acids in butter. There was a considerable recirculation of ALA into the NEFA pool available for oxidation. Practical application: Enzymatic transesterification was used to produce a dietary oil rich in ALA. By randomizing the partitioning of ALA more evenly between the TAG molecules the risk of oxidation could be reduced. Analyses showed that the ALA-oil was stable during storage for at least 3 months. Enzymatic transesterification could be used as an advantageous method to design an ALA rich dietary oil with new properties regarding fatty acid composition, susceptibility to oxidation, and effects on blood lipids. [source] Cloning of Xenopus orthologs of Ctf7/Eco1 acetyltransferase and initial characterization of XEco2FEBS JOURNAL, Issue 24 2008Masatoshi Takagi Sister chromatid cohesion is important for the correct alignment and segregation of chromosomes during cell division. Although the cohesin complex has been shown to play a physical role in holding sister chromatids together, its loading onto chromatin is not sufficient for the establishment of sister chromatid cohesion. The activity of the cohesin complex must be turned on by Ctf7/Eco1 acetyltransferase at the replication forks as the result of a specific mechanism. To dissect this mechanism in the well established in vitro system based on the use of Xenopus egg extracts, we cloned two Xenopus orthologs of Ctf7/Eco1 acetyltransferase, XEco1 and XEco2. Both proteins share a domain structure with known members of Ctf7/Eco1 family proteins. Moreover, biochemical analysis showed that XEco2 exhibited acetyltransferase activity. We raised a specific antibody against XEco2 and used it to further characterize XEco2. In tissue culture cells, XEco2 gradually accumulated in nuclei through the S phase. In nuclei formed in egg extract, XEco2 was loaded into the chromatin at a constant level in a manner sensitive to geminin, an inhibitor of the pre-replication complex assembly, but insensitive to aphidicolin, an inhibitor of DNA polymerases. In both systems, no specific localization was observed during mitosis. In XEco2-depleted egg extracts, DNA replication occurred with normal kinetics and efficiency, and the condensation and sister chromatid cohesion of subsequently formed mitotic chromosomes was unaffected. These observations will serve as a platform for elucidating the molecular function of Ctf7/Eco1 acetyltransferase in the establishment of sister chromatid cohesion in future studies, in which XEco1 and XEco2 should be dissected in parallel. [source] Simultaneous detection of the establishment of seed-inoculated Pseudomonas fluorescens strain DR54 and native soil bacteria on sugar beet root surfaces using fluorescence antibody and in situ hybridization techniquesFEMS MICROBIOLOGY ECOLOGY, Issue 1 2000Peter Stephensen Lübeck Abstract Colonization at sugar beet root surfaces by seedling-inoculated biocontrol strain Pseudomonas fluorescens DR54 and native soil bacteria was followed over a period of 3 weeks using a combination of immunofluorescence (DR54-targeting specific antibody) and fluorescence in situ hybridization (rRNA-targeting Eubacteria EUB338 probe) techniques with confocal laser scanning microscopy. The dual staining protocol allowed cellular activity (ribosomal number) to be recorded in both single cells and microcolonies of strain DR54 during establishment on the root. After 2 days, the population density of strain DR54 reached a constant level at the root basis. From this time, however, high cellular activity was only found in few bacteria located as single cells, whereas all microcolony-forming cells occurring in aggregates were still active. In contrast, a low density of strain DR54 was observed at the root tip, but here many of the bacteria located as single cells were active. The native population of soil bacteria, comprising a diverse assembly of morphologically different forms and size classes, initiated colonization at the root basis only after 2 days of incubation. Hence the dual staining protocol allowed direct microscopic studies of early root colonization by both inoculant and native soil bacteria, including their differentiation into active and non-active cells and into single or microcolony-forming cells. [source] Adaptive control for non-negative and compartmental dynamical systems with applications to general anesthesiaINTERNATIONAL JOURNAL OF ADAPTIVE CONTROL AND SIGNAL PROCESSING, Issue 3 2003Wassim M. Haddad Abstract Non-negative and compartmental dynamical system models are composed of homogeneous interconnected subsystems or compartments which exchange variable non-negative quantities of material with conservation laws describing transfer, accumulation, and elimination between the compartments and the environment. These models are widespread in biological and physiological sciences and play a key role in understanding these processes. In this paper, we develop a direct adaptive control framework for linear uncertain non-negative and compartmental systems. The proposed framework is Lyapunov-based and guarantees partial asymptotic set-point regulation; that is, asymptotic set-point stability with respect to part of the closed-loop system states associated with the plant. In addition, the adaptive controller guarantees that the physical system states remain in the non-negative orthant of the state space. Finally, a numerical example involving the infusion of the anesthetic drug propofol for maintaining a desired constant level of depth of anesthesia for non-cardiac surgery is provided to demonstrate the efficacy of the proposed approach. Copyright © 2003 John Wiley & Sons, Ltd. [source] KINETICS OF ACRYLAMIDE FORMATION DURING TRADITIONAL AND VACUUM FRYING OF POTATO CHIPSJOURNAL OF FOOD PROCESS ENGINEERING, Issue 5 2005CLAUDIA GRANDA ABSTRACT Acrylamide is considered a carcinogen in animals and a possible carcinogen in humans. It has been found in starch-rich foods cooked at high temperatures. Vacuum frying (10 Torr) was investigated as a possible alternative to reduce acrylamide formation in potato chips. The cultivar Atlantic was used to determine the kinetics of acrylamide formation during traditional and vacuum frying at different temperatures. There was a 94% decrease in acrylamide content when potatoes were fried to the same final moisture content (1.5% ± 0.3% w.b.) under vacuum compared to those fried under atmospheric conditions. Acrylamide accumulation under vacuum frying was modeled using first-order kinetics (during traditional frying, the logistic kinetic model was used). The behavior of the kinetics of acrylamide content in potato chips fried under the two processes was different mainly because of the different temperatures used. During traditional frying, higher temperatures are used (150 to 180C) and acrylamide after some time is produced but starts degrading, producing a constant level of acrylamide content at longer times. During vacuum frying (10 Torr), acrylamide increased exponentially (but at lower levels) for all frying times. [source] Age Determination of Ballpoint Pen Ink by Thermal Desorption and Gas Chromatography,Mass Spectrometry,JOURNAL OF FORENSIC SCIENCES, Issue 4 2008Jürgen H. Bügler Ph.D. Abstract:, Two main approaches can be used for determining the age of an ink: indirect dating and direct dating. Indirect dating is based on the chemical analysis of an ink followed by comparison with known samples in a reference collection. The collection should contain information about the inks including the market introduction dates. This approach may allow for an anachronism to be detected. The second concept is based on measuring ink components that change with age. The analysis of solvents in ballpoint inks may be a useful parameter for determining the age of ink on paper. In a previous study, the authors demonstrated that thermal desorption of ink directly from paper, followed by chemical analysis using gas chromatography,mass spectrometry (GC,MS), is a promising procedure for characterizing ink-binder resins and solvents. Preliminary tests showed that monitoring the evaporation of ink solvent from ink on paper is not a suitable method for ink dating. Thermal analysis of ink on paper in two steps revealed that fresh ink releases a relative amount of solvent at a certain low temperature in a defined period of time, which decreases as the ink ages. As a consequence, this relative amount of solvent released at a certain low temperature, and its decrease with time, can be used to estimate ink age. This age-dependent parameter was studied in 85 different inks ranging in age from 1 week to 1.5 years. It was found that some inks showed a significant decrease of this parameter up to an age of several months, and that the aging process can be monitored within this period. For other inks, however, the age-dependent parameter decreases relatively fast, e.g., within a few days, to a constant level, which can be too fast for casework. Based on these results, a general procedure for assessing the age of ballpoint pen inks on paper was developed. [source] Low-dose prostacyclin in treatment of severe brain trauma evaluated with microdialysis and jugular bulb oxygen measurementsACTA ANAESTHESIOLOGICA SCANDINAVICA, Issue 7 2000P.-O. Grände Background: The endogenous substance prostacyclin is a substance with the potential to improve microcirculation and oxygenation around contusions in the brain following a head trauma by its vasodilatory, antiaggregatory and antiadhesive effects. Microdialysis measurements of local concentrations of selected interstitial substances in the brain, and measurements of venous jugular bulb oxygenation reflecting overall brain oxygenation, might be useful to evaluate possible therapeutic effects of a specific therapy, such as treatment with prostacyclin. Methods: This case report study on six patients, of whom five were given prostacyclin, includes cerebral microdialysis measurements of interstitial lactate (n=5), pyruvate (n=3), glycerol (n=5) and glucose (n=4), and is combined with measurements of venous jugular bulb oxygenation in three of the patients. One microdialysis catheter was placed adjacent to a contusion, and in four of the patients another catheter was also placed in the contralateral less injured side for comparison. Low-dose prostacyclin infusion (0.5,1.0 ng kg,1 min,1) was started when lactate concentrations in the more injured side was raised at a constant level for more than 10 h. The study also includes one patient used as control to whom no prostacyclin was given. Results: Lactate was markedly lower in the less injured than in the more injured area of the brain. During the prostacyclin infusion elevated lactate and lactate/pyruvate ratio were reduced. Elevated glycerol decreased, a low glucose increased and jugular bulb blood oxygenation increased following start of prostacyclin. The control patient showed an increase in lactate and lactate/pyruvate ratio. Conclusion: The microdialysis data combined with the jugular bulb oxygenation data indicated that low-dose prostacyclin exerts effects compatible with improved oxygenation and reduced cell damage in the severely traumatised brain. [source] Fruit load and elevation affect ethylene biosynthesis and action in apple fruit (Malus domestica L. Borkh) during development, maturation and ripeningPLANT CELL & ENVIRONMENT, Issue 11 2007VALERIANO DAL CIN ABSTRACT The influence of internal and external factors such as tree fruit load and elevation on ethylene biosynthesis and action was assessed during apple fruit development and ripening. Ethylene biosynthesis, as well as transcript accumulation of the hormone biosynthetic enzymes (MdACS1 and MdACO1), receptors (MdETR1 and MdERS1) and an element of the transduction pathway (MdCTR1), were evaluated in apples borne by trees with high (HL) and low (LL) fruit load. Orchards were located in two localities differing in elevation and season day degree sum. These parameters significantly affected the date of bloom and commercial harvest, and the length of the fruit developmental cycle. Trees from the low elevation (LE) bloomed and the fruit ripened earlier than those from the high elevation (HE), displaying also a shortened fruit developmental cycle. Dynamics of ethylene evolution was apparently not affected by elevation. The onset of ethylene evolution started 130 days after bloom (DAB) at both elevations. During early ripening, fruits from LL trees produced significantly more ethylene than those from HL trees. Expression analysis of MdACS1, MdACO1 and MdERS1 indicated that the transcript accumulation well correlated with ethylene evolution. MdCTR1 was expressed at constant level throughout fruit growth and development up to 130 DAB, thereafter, the transcript accumulation decreased up to commercial harvest, concurrently with the onset of ethylene evolution. [source] An optimal test against a random walk component in a non-orthogonal unobserved components modelTHE ECONOMETRICS JOURNAL, Issue 2 2002Ralph W. Bailey Summary In this paper we consider the problem of testing the null hypothesis that a series has a constant level (possibly as part of a more general deterministic mean) against the alternative that the level follows a random walk. This problem has previously been studied by, inter alia, Nyblom and Mäkeläinen (1983) in the context of the orthogonal Gaussian random walk plus noise model. This model postulates that the noise component and the innovations to the random walk are uncorrelated. We generalize their work by deriving the locally best invariant test of a fixed level against a random walk level in the non-orthogonal case. Here the noise and random walk components are contemporaneously correlated with correlation coefficient ,. We demonstrate that the form of the optimal test in this setting is independent of ,; i.e. the test statistic previously derived for the case of ,= 0 remains the locally optimal test for all ,. This is a very useful result: it states that the locally optimal test may be achieved without prior knowledge of ,. Moreover, we show that the limiting distribution of the resulting statistic under both the null and local alternatives does not depend on ,, behaving exactly as if ,= 0. Finite sample simulations of these effects are provided to illustrate and generalizations to models with dependent errors are considered. [source] Treatment with leuprolide acetate decreases the threshold of the ventilatory response to carbon dioxide in healthy malesTHE JOURNAL OF PHYSIOLOGY, Issue 2 2004Jason H. Mateika This investigation was designed to determine if suppression of testosterone alters the ventilatory response to carbon dioxide in the presence of high and low levels of oxygen. Eleven healthy male subjects completed a series of rebreathing trials during wakefulness, before and after treatment with a long-acting gonadotropin-releasing hormone agonist. Five subjects also completed studies during non-rapid eye movement (NREM) sleep. During wakefulness, subjects initially hyperventilated to reduce the partial pressure of carbon dioxide (PET,CO2) below 25 Torr. Subjects then rebreathed from a bag containing a normocapnic (42 Torr), low (50 Torr) or high oxygen (140 Torr) gas mixture. During each trial PET,CO2 increased while oxygen was maintained at a constant level. The threshold of the ventilatory response to carbon dioxide was considered to be the point at which minute ventilation began to rise in a linear fashion as PET,CO2 increased. The slope of the ventilatory response above the threshold was used as a measure of sensitivity to carbon dioxide. During NREM sleep, hypocapnia was induced via nasal mechanical ventilation. Several trials were completed until the cessation of mechanical ventilation resulted in a central apnoea which demarcated the threshold of the ventilatory response to carbon dioxide. In response to treatment with leuprolide acetate, the threshold measured in wakefulness decreased during carbon dioxide rebreathing in the presence of low (41.05 ± 0.77 versus 39.40 ± 0.83 Torr; P= 0.01) and high (46.32 ± 0.56 versus 44.78 ± 0.83 Torr; P= 0.01) oxygen levels. An increase in sensitivity (4.82 ± 0.61 versus 7.17 ± 1.20 l min,1 Torr,1; P= 0.02) was also observed during rebreathing in the presence of high but not low oxygen levels. The increase in sensitivity was accompanied by an increase in carbon dioxide production. The findings observed during NREM sleep were similar to those observed during wakefulness, since the PET,CO2 that demarcated the threshold was decreased after leuprolide treatment (42.1 ± 0.6 versus 39.6 ± 0.6 Torr; P= 0.002). Additionally, the decrease in PET,CO2 required to induce an apnoea was greater after treatment with leuprolide (2.56 ± 0.25 versus 4.06 ± 0.29 Torr; P= 0.004). We conclude that suppression of testosterone decreases the threshold of the ventilatory response to carbon dioxide during both wakefulness and sleep. [source] Antimicrobial resistance 1979,2009 at Karolinska hospital, Sweden: normalized resistance interpretation during a 30-year follow-up on Staphylococcus aureus and Escherichia coli resistance developmentAPMIS, Issue 9 2010GÖRAN KRONVALL Kronvall G. Antimicrobial resistance 1979,2009 at Karolinska hospital, Sweden: normalized resistance interpretation during a 30-year follow-up on Staphylococcus aureus and Escherichia coli resistance development. APMIS 2010; 118: 621,39. To utilize a material of inhibition zone diameter measurements from disc diffusion susceptibility tests between 1979 and 2009, an objective setting of epidemiological breakpoints was necessary because of methodological changes. Normalized resistance interpretation (NRI) met this need and was applied to zone diameter histograms for Staphylococcus aureus and Escherichia coli isolates. The results confirmed a slow resistance development as seen in Northern countries. The S. aureus resistance levels for erythromycin, clindamycin and fusidic acid in 2009 were 3.2%, 1.8% and 1.4% with denominator correction. A rise in resistance to four antimicrobials in 1983 was probably because of a spread of resistant Methicillin Susceptible Staphylococcus Aureus (MSSA). For E. coli, the denominator-corrected resistance levels in 2009 were 27% for ampicillin, around 3% for third-generation cephalosporins, 0.1% for imipenem, 2.5% for gentamicin, 19% for trimethoprim, 4.5% for co-trimoxazole, 1.2% for nitrofurantoin and 9% for ciprofloxacin. The temporal trends showed a rise in fluoroquinolone resistance from 1993, a parallel increase in gentamicin resistance, a substantial increase in trimethoprim and sulphonamide resistance in spite of decreased consumption, and a steady rise in ampicillin resistance from a constant level before 1989. A short review of global resistance surveillance studies is included. [source] Postnatal development of ,-cells in ratsAPMIS, Issue 5 2002Proposed explanatory model The previously shown wave of ,-cell apoptosis and the apparent plateau in the ,-cell mass in the third week of life in rats are still unexplained events. Using a novel design-based stereological method we investigated the postnatal development of the ,-cell population in Sprague-Dawley rats. The total ,-cell mass increased from postnatal day 4 until day 16, to be followed by a plateau until day 24, after which it increased further. This plateau was caused by ,-cell hypotrophia as well as decreased net ,-cell formation. The ,-cell mass per unit body weight (the relative ,-cell mass) was five times higher at birth compared with the adult constant level that was reached at approximately 24 days of age. We propose an explanatory model for the postnatal development of the ,-cell population in rats. According to this model, ,-cells in the early postnatal period are immature, i.e. are not susceptible to the mechanism that in later life maintains a constant relative ,-cell mass. Within the following weeks the number of mature ,-cells increases, and from approximately day 24 and onwards the ,-cell population is dominated by mature ,-cells that adjust to match the body weight, keeping a constant relative ,-cell mass. Findings of an apoptotic wave, a plateau phase in the total ,-cell mass development, a period with ,-cell hypotrophia, and the disappearance of insulin-like growth factor II positive ,-cells at postnatal day 21 all fit well in the model. [source] Cobalt-, zinc- and iron-bound forms of adenylate kinase (AK) from the sulfate-reducing bacterium Desulfovibrio gigas: purification, crystallization and preliminary X-ray diffraction analysisACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 9 2009A. V. Kladova Adenylate kinase (AK; ATP:AMP phosphotransferase; EC 2.7.4.3) is involved in the reversible transfer of the terminal phosphate group from ATP to AMP. AKs contribute to the maintenance of a constant level of cellular adenine nucleotides, which is necessary for the energetic metabolism of the cell. Three metal ions, cobalt, zinc and iron(II), have been reported to be present in AKs from some Gram-negative bacteria. Native zinc-containing AK from Desulfovibrio gigas was purified to homogeneity and crystallized. The crystals diffracted to beyond 1.8,Å resolution. Furthermore, cobalt- and iron-containing crystal forms of recombinant AK were also obtained and diffracted to 2.0 and 3.0,Å resolution, respectively. Zn2+,AK and Fe2+,AK crystallized in space group I222 with similar unit-cell parameters, whereas Co2+,AK crystallized in space group C2; a monomer was present in the asymmetric unit for both the Zn2+,AK and Fe2+,AK forms and a dimer was present for the Co2+,AK form. The structures of the three metal-bound forms of AK will provide new insights into the role and selectivity of the metal in these enzymes. [source] Measurement of key metabolic enzyme activities in mammalian cells using rapid and sensitive microplate-based assaysBIOTECHNOLOGY & BIOENGINEERING, Issue 3 2010R. Janke Abstract Sensitive microplate-based assays to determine low levels of key enzyme activities in mammalian cells are presented. The enzyme platform consists of four cycling assays to measure the activity of 28 enzymes involved in central carbon and glutamine metabolism. The sensitivity limit of all cycling assays was between 0.025 and 0.4,nmol product. For the detection of glutaminase activity, a new glutamate cycle system involving the enzymes glutamate dehydrogenase and aspartate transaminase was established. The relative standard deviation of the method was found to be 1.7% with a limit of detection of 8.2,pmol and a limit of quantitation of 24.8,pmol. Hence, cell extracts could be highly diluted to reduce interferences caused by other components in the extract, which in addition minimized underestimates or overestimates of actual enzyme activities. Since substrate concentrations could be maintained at a nearly constant level throughout the assay product accumulation during the reaction was low, which minimized product inhibition. As an example, the enzyme platform was used to investigate maximum enzyme activities of stationary-phase MDCK cells grown in serum-containing GMEM medium as typically used in influenza vaccine production. Biotechnol. Bioeng. 2010;107: 566,581. © 2010 Wiley Periodicals, Inc. [source] The impact of salinity pulses on the emergence of plant and zooplankton from wetland seed and egg banksFRESHWATER BIOLOGY, Issue 5 2007DARYL L. NIELSEN Summary 1. In this study we compared the emergence of aquatic biota from sediments under 14-day pulses of high (5000 mg L,1) and low (1000 mg L,1) salinity with emergence under freshwater and equivalent constant salinity levels. We tested the hypothesis that pulses of high salinity and short duration have no impact on the emergence of aquatic plants and zooplankton from wetland sediment. 2. The way salt is moved through the landscape may alter the response of biota to increases in salinity. Under natural hydrological regimes in rivers and floodplains salinity pulses occur often at concentrations that exceed predicted tolerance levels for aquatic biota. The impacts of natural pulses of high salinity followed by rapid return to fresh conditions may be used to inform management guidelines for the potential release of non-natural saline water into river systems with minimal impact. 3. For both aquatic plants and zooplankton the abundance and richness of the emerging taxa decreased at higher salinities kept at constant levels. In contrast, pulses of salinity followed by return to freshwater conditions did not have a negative impact on the emergence of aquatic plants or zooplankton. For many taxa of zooplankton a positive impact was demonstrated with higher emergence following the salinity pulse. 4. The responses of aquatic plant and zooplankton taxa are grouped into five response types. Type 1: negatively impacted by all salt regimes. Type 2: preference for constant salinities. Type 3: no difference between fresh and either pulse regime. Type 4: preference for high concentration pulses. Type 5: emergence higher under a low concentration pulse. 5. Although previous studies indicate that constant high-level salinity in rivers and wetlands can decrease the species richness of aquatic communities, this current study shows pulses may not have the same impact. Our results support the hypothesis that pulses of high salinity and short duration do not impact on the emergence of aquatic plants and zooplankton from wetland sediments. For zooplankton, pulses of salt may trigger emergence. 6. These trends may be used to explore the potential to use managed water releases to move salt through the landscape with minimal impact of salinity on aquatic biota. However, before such preliminary results are applied in management of saline water releases we need to determine the implications for interacting processes in natural ecosystems. [source] Ontogeny of human hepatic cytochromes P450JOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 4 2007Ronald N. Hines Abstract Significant changes in drug-metabolizing enzyme (DME) expression occur during ontogeny. Such changes can have a profound effect on therapeutic efficacy in the fetus and child, as well as the risk for adverse drug reactions. To gain a better understanding of DME ontogeny, enzyme contents for six key cytochromes P450 were measured in 240 human liver samples representing ages from 8 weeks gestation to 18 years. Where possible, both quantitative western blotting and activity assays with probe substrates were performed. Although oversimplified, the DME can be grouped into one of three categories. As typified by CYP3A7, some enzymes are expressed at their highest level during the first trimester and either remain at high concentrations or decrease during gestation and are silenced or expressed at low levels within 1,2 years after birth. These data cause one to query whether these enzymes have an important endogenous function. Representatives of a second group, CYP3A5 and CYP2C19, are expressed at relatively constant levels throughout gestation. Postnatal increases in CYP2C19 are observed within the first year, but not for CYP3A5. CYP2C9, 2E1, and 3A4 are more typical of a third group of enzymes that are not expressed or are expressed at low levels in the fetus with the onset of expression generally in either the second or third trimester. Substantial increases in expression are observed within the first 1,2 years after birth; however, considerable interindividual variability is observed in the immediate postnatal (1,6 months) onset or increase in expression of these enzymes, often resulting in a window of hypervariability. © 2007 Wiley Periodicals, Inc. J Biochem Mol Toxicol 21:169,175, 2007; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20179 [source] Expression of signal transduction proteins during the differentiation of primary human erythroblastsJOURNAL OF CELLULAR PHYSIOLOGY, Issue 3 2005Viviana di Giacomo The high number (>108,10) of primary human pro-erythroblasts (CD36high/CD235alow) obtainable in HEMA culture (Migliaccio et al., 2002) is exploited here to analyse the expression of proteins implicated in erythropoietin (EPO)-signalling (STATs, PI-3K, and PLCs) during the process of erythroid maturation. Human pro-erythroblasts progressed in 4 days of culture with EPO into basophilic- (CD36high/CD235amedium, 24 h), polychromatic-(CD36high/CD235ahigh, 48 h), and, finally, orthochromatic-(CD36low/CD235ahigh, 72,96 h) erythroblasts. During this maturation, STAT-1 was expressed up to the orthochromatic stage, expression of STAT-5, as well as of its target proteins BclxL and IRF1, remained constant up to 48 h (polychromatic-erythroblasts) but decreased by 96 h (orthochromatic-erythroblasts), while that of STAT-3 decreased constantly from 24 h on and became undetectable by 96 h. Expression of PI-3K rapidly decreased with differentiation since only 50% of original protein levels were detected by 48 h. On the other hand, among the members of PLC families investigated, PLC ,4 was not expressed, PLC ,2, ,1, and ,2 were expressed at constant levels throughout the maturation process, while expression of PLC ,3 and of PLC ,1 decreased, as PI-3K, by 24 h and that of PLC ,1 was induced by 6 h and became undetectable by 24 h. In conclusion, these data depict the dynamic signalling scenario associated with the maturation of erythroid cells and provide the first indication that members of PLC families (PLC ,1, ,3, and ,1) might e involved in the control of erythroid differentiation in humans. © 2004 Wiley-Liss, Inc. [source] Crossing the barrier: oxysterols as cholesterol transporters and metabolic modulators in the brainJOURNAL OF INTERNAL MEDICINE, Issue 6 2006I. BJÖRKHEM Abstract. A normal brain function requires constant levels of cholesterol, and the need for constancy seems to be higher here than in any other organ. Nature has met this need by isolation of brain cholesterol by a highly efficient blood,brain barrier. As a low synthesis of cholesterol is present in the brain, a mechanism for compensatory elimination is required. A decade ago we made the unexpected finding that the favoured mechanism for this involves conversion into 24S-hydroxycholesterol, followed by diffusion over the blood,brain barrier. Recent studies by us and others on this new pathway have given new insights into the mechanisms by which cholesterol homeostasis is maintained in the brain. We recently demonstrated a flux of another oxygenated product of cholesterol, 27-hydroxycholesterol, in the opposite direction. The latter flux may be important for neurodegeneration, and may be the link between hypercholesterolaemia and Alzheimer's disease. An overview of the above studies is presented and the possibility that the cholesterol 24S-hydroxylase in the brain may be important for memory and learning and that it may be a new drug target is discussed. [source] Pharmaceutical analysis by supercritical fluid chromatography: Optimization of the mobile phase composition on a 2-ethylpyridine columnJOURNAL OF SEPARATION SCIENCE, JSS, Issue 8 2008Claudio Brunelli Abstract The separation of neutral, acidic, and basic pharmaceuticals with diverse physicochemical properties by packed column supercritical fluid chromatography (pSFC) on a 2-ethylpyridine column (25 cm×4.6 mm id, 3 ,m particles) is presented. The optimization strategy involved separations at 100% methanol (MeOH) and at 50% MeOH/50% ACN while keeping the peak symmetry additives formic acid (FA) and isopropylamine (IPA) at constant levels of 0.25% v/v. By plotting the adjusted retention times as a function of the MeOH/ACN ratio, an optimal modifier ratio composition of 65% MeOH/35% ACN was found. The total set of 26 neutral, acidic, and basic pharmaceuticals was analyzed and the optimal composition experimentally verified. This mobile phase composition is currently used in pharmaceutical method development and open-access generic screening environments. [source] Comparative dry matter intake and nutrient utilisation efficiency in lactating cattle and buffaloesJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 4 2003Shyam S Paul Abstract An attempt was made to assess the comparative dry matter intake and nutrient utilisation efficiency of lactating buffaloes and cows based on results obtained from experimental feeding trials conducted in India. Data on dry matter (DM) intake, total digestible nutrient (TDN) intake, digestible crude protein (DCP) intake, body weight or metabolic body size (MBS), body weight change, milk yield or 4% fat-corrected milk (FCM) yield, milk fat percentage and roughage/concentrate ratio in the diet of lactating buffaloes and cows were collected from published reports. The data were processed and analysed to assess the comparative dry matter intake and nutrient utilisation efficiency using suitable statistical analysis models. DM intake was significantly (P,<,0.01) lower in buffaloes (2.57,kg DM per 100,kg body weight or 119.2,g,kg,1 MBS) than in cattle (3.09,kg DM per 100,kg body weight or 132.0,g,kg,1 MBS). Mean gross energetic efficiency (30.53 versus 27.83%; P,<,0.01), gross protein efficiency (45.48 versus 37.06%; P,<,0.01), net energetic efficiency (69.16 versus 64.10%; P,<,0.05) and net protein efficiency (80.15 versus 59.59%; P,<,0.001) were significantly higher in buffaloes than in cattle. Lactating buffaloes consumed significantly less (P,<,0.001) protein (75.69,g DCP) and less (P,<,0.01) energy (695.9,g TDN) than cows (93.89,g DCP, 774.8,g TDN) for production of 1,kg of 4% fat-corrected milk. Buffaloes had higher energy and protein utilisation efficiencies as compared with cattle at similar fat-corrected milk production level, plane of energy and protein nutrition, body size and body weight change. Buffaloes (1.189,kg DM,kg,1 4% FCM) consumed a similar (P,>,0.05) amount of feed dry matter to that of cows (1.267,kg DM,kg,1 4% FCM). However, when DM intake,kg,1 FCM (4%) was compared at constant levels of metabolic body size, fat-corrected milk, body weight change, dietary energy concentration and green forage percentage in the diet, lactating buffaloes consumed significantly less DM,kg,1 FCM yield as compared with cattle. It was concluded that DM intake was lower in lactating buffaloes. Moreover, lactating buffaloes utilised dietary dry matter, energy and protein for milk production more efficiently than cattle. © 2003 Society of Chemical Industry [source] Inactivation of Virulent Burkholderia pseudomallei by SunlightPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2009Jose-Luis Sagripanti The goal of this study was to determine the sensitivity of virulent Burkholderia pseudomallei to natural sunlight. We describe solar dosimetry calibrated to integrate radiation between 295 and 305 nm and an exposure system that minimizes thermal effects on bacterial cells. Burkholderia pseudomallei cells were either exposed to sunlight in UV transparent dishes or maintained in the dark covered by opaque foil. The cells maintained in the dark remained at constant levels for the duration of all experiments. The exposed cells nearby were killed with a kinetic studied through 5 Log10 inactivation. We found that cells in stationary phase of growth were nearly two-fold more resistant to sunlight than cells in lag or exponential growth. A virulent strain of B. pseudomallei that produced mucoid colonies showed sensitivity to sunlight similar to both a virulent strain that produced nonmucoid colonies and a strain of B. thailandensis. The inactivation of B. pseudomallei by sunlight in different types of water of environmental relevance or inside amoebae was investigated. The sensitivity of virulent B. pseudomallei was calculated and its comparison with previous studies employing monochromatic germicidal light (254 nm) is discussed. This may be the first report in the open literature of the inactivation of a virulent biological threat agent by natural sunlight. These data should assist in estimating the risk for contracting melioidosis and in predicting the time period during which B. pseudomallei remains infectious after an accidental or intentional release in the environment. 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