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  • gene expression  (69)
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  • 04. Solid Earth::04.06. Seismology::04.06.05. Historical seismology
  • 550 - Earth sciences
  • conservation
  • Springer  (167)
  • Deutsches GeoForschungsZentrum GFZ  (13)
  • Blackwell Publishing Ltd
  • Community Online Resource for Statistical Seismicity Analysis
  • Deutsches GeoForschungsZentrum
  • Essen : Verl. Glückauf
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  • 2005-2009
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  • 1995-1999  (180)
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  • 1
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    Deutsches GeoForschungsZentrum GFZ
    In:  Scientific Technical Report
    Publication Date: 2020-02-12
    Description: Die vorliegende Arbeit präsentiert ein quantitatives tektonisches Modell für das NW' saxothuringische Becken, das im NW-Sporn der Böhmischen Masse aufgeschlossen ist (Thüringer Wald und Frankenwald). Die Untersuchungen konzentrieren sich auf eine Profiltraverse von der Vesser-Zone im NW bis zur Frontüberschiebung des Münchberger Deckenstapels im SE, zwischen Fränkischer Linie im SW und Frankenwälder Querzone im NE. Der zugrundeliegende geologisch-geophysikalische Datensatz basiert zu großen Teilen auf publizierten Befunden, ergänzt durch eigene strukturgeologische Untersuchungen und geothermobarometrische Abschätzungen der syntektonischen Metamorphosebedingungen.
    Keywords: 550 - Earth sciences
    Language: German
    Type: info:eu-repo/semantics/doctoralThesis
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  • 2
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    Deutsches GeoForschungsZentrum GFZ
    In:  Scientific Technical Report STR
    Publication Date: 2022-11-30
    Keywords: 550 - Earth sciences
    Language: German
    Type: info:eu-repo/semantics/report
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  • 3
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    Deutsches GeoForschungsZentrum GFZ
    In:  Scientific Technical Report STR
    Publication Date: 2022-11-30
    Keywords: 550 - Earth sciences
    Language: German
    Type: info:eu-repo/semantics/report
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  • 4
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
    Type: info:eu-repo/semantics/report
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  • 5
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
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  • 6
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    Deutsches GeoForschungsZentrum GFZ
    In:  Scientific Technical Report STR
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
    Type: info:eu-repo/semantics/doctoralThesis
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  • 7
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    Deutsches GeoForschungsZentrum GFZ
    In:  Scientific Technical Report STR
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
    Type: info:eu-repo/semantics/doctoralThesis
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  • 8
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
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  • 9
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    Deutsches GeoForschungsZentrum GFZ
    In:  Scientific Technical Report STR
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
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  • 10
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    Deutsches GeoForschungsZentrum GFZ
    In:  Scientific Technical Report STR
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
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  • 11
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
    Type: info:eu-repo/semantics/doctoralThesis
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  • 12
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    Deutsches GeoForschungsZentrum GFZ
    In:  Scientific Technical Report STR
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
    Type: info:eu-repo/semantics/doctoralThesis
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  • 13
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    Deutsches GeoForschungsZentrum GFZ
    In:  Scientific Technical Report STR
    Publication Date: 2022-11-14
    Keywords: 550 - Earth sciences
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  • 14
    Electronic Resource
    Electronic Resource
    Springer
    European journal of nutrition 36 (1997), S. 299-302 
    ISSN: 1436-6215
    Keywords: Energy retention ; protein retention ; fat retention ; growth ; body composition ; broilers ; Energieansatz ; Proteinansatz ; Fettansatz ; Wachstum ; Körperzusammensetzung ; Broiler
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Zusammenfassung Mit steigender Aufnahme erhöht sich der Körperfettgehalt und der Körperproteingehalt nimmt ab. Es wird oft angenommen, daß jede Zunahme im Ansatz mit mehr Fett und weniger Protein verbunden ist. Experimentelle Ergebnisse widerlegen jedoch diese Annahme. In zwei Experimenten mit männlichen Broilerhühnern wurden die Tiere auf einem Niveau von 60 % und 100 % der empfohlenen Energieaufnahme gefüttert. Die Körperzusammensetzung bei 1500 g zeigte, wie erwartet, daß sich bei steigenden Aufnahmen der Körperfettgchalt vergrößert und der Körperproteingehalt abnimmt. Fettund Proteinansatz waren linear mit der totalen Energieretention (ER) korreliert. Das bedeutet, daß jede Zunahme im Ansatz den gleichen Protein- und Fettgehalt besitzt. Wenn der Fettansatz gleich Null ist wird nur Protein, etwa 50 % des maximalen Ansatzes, retiniert. Wenn ER=O ist, wird Protein angesetzt und Fett mobilisiert. Energie-und N-Bilanzuntersuchungen bestätigen die konstante Zusammensetzung jeder Vergrößerung des Ansatzes. Die Ergebnisse beider Experimente zeigen, daß die ER aus zwei Komponenten besteht: einem basalen konstanten täglichen Proteinansatz und einer variablen zusätzlichen ER, die hauptsächlich aus Fett besteht. Der basale Proteinansatz beträgt etwa 50 % des maximalen Ansatzes. Mit steigenden Energieaufnahmen wird der basale Proteinansatz mit einer zusätzlichen Menge von Protein und Fett im konstanten Verhältnis ergänzt.
    Notes: Summary With increasing intakes the body fat content increases and that of protein decreases. It is most often assumed that this is brought about because each increment in retention contains more fat and less protein. Experimental results, however, showed that this explanation is not true. In two experiments male broiler chickens were fed at levels between 60 and 100 % of recommended energy intake. Body composition at 1500 g showed, as expected, that with increasing intakes body fat content increased and protein content decreased. Both fat and protein retention per day were linearly related to total energy retention (ER). This means that each increment in retention has the same protein and fat content. At zero fat retention only protein was retained, about 50 % of maximal retention. At zero ER protein was retained and fat mobilized. Energy and N balance experiments confirmed the constant composition of each increment in retention. The results of both experiments show that total ER consisted of two components: a basic constant daily protein retention and a variable additional ER, mainly consisting of fat. The basic protein retention is about half of maximal retention. With increasing energy intakes the basic protein retention is combined with an additional amount of protein and fat in a constant ratio.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    European journal of nutrition 36 (1997), S. 332-335 
    ISSN: 1436-6215
    Keywords: Nutrient intake ; protein metabolism ; protein synthesis ; growth ; energy expenditures ; Nährstoffaufnahme ; Proteinstoffwechsel ; Proteinsynthese ; Wachstum ; Energieaufwand
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Zusammenfassung Eine Zunahme von fettfreiem Gewebe tritt auf, wenn die Proteinsynthese größer ist als der Proteinabbau. Obwohl während des Wachstums von der Geburt bis zur Reife die absoluten Proteinsynthese- und -abbauraten ansteigen, nehmen dagegen die fraktionellen Raten ab. Bcide Prozesse reagieren auf die Nährstoffaufnahme. Es gibt aber deutliche Unterschiede zwischen den verschiedenen Geweben. Protein, Kohlenhydrate und Fett können den Proteinansatz bei unreifen Tieren und Kindern stimulieren. Die zugrundeliegenden Mechanismen und die Energieaufwendungen scheinen jedoch unterschiedlich zu sein.
    Notes: Summary Lean tissue growth occurs when the rate of protein synthesis exceeds the rate of protein breakdown. Althoughabsolute rates of protein synthesis and breakdown rise during growth from birth to maturityfractional rates fall. Both these processes are sensitive to nutrient intake but responses to feeding vary greatly amongst different tissues. Protein, carbohydrate and fat can all stimulate body protein accretion in immature animals and in children but the mechanisms by which they do so, and the energy expenditures involved, seem to be different.
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  • 16
    Electronic Resource
    Electronic Resource
    Springer
    European journal of nutrition 36 (1997), S. 205-213 
    ISSN: 1436-6215
    Keywords: Frying ; growth ; liver lipids ; lipaemia ; olive oil ; pregnancy ; rat ; Fritierung ; Wachstum ; Leberfett ; Olivenöl ; Schwangerschaft ; Ratte
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Zusammenfassung Um Informartionen über den Zusammenhang zwischen der Aufnahme von Fett, das zum Fritieren benutzt wurde, und dem Fetthaushalt während Perioden starken Körperaufbaus zu haben, wurde der Einfluß der Aufnahme von frischem Olivenöl (Gehalt polarer Verbindungen, 2 %; Ölsäure 78,9 mg/100 mg Öl, und Linolsäure 7 mg/100 mg Öl) und von Olivenöl, das 15 mal in Folge für das Fritieren von Kartoffeln benutzt worden war (Gehalt polarer Verbindungen 9 %; Ölsäure 75,8 mg/100mg Öl und Linolsäure 6,2 mg/100mg Öl) während der Gravidität, untersucht. Dazu wurden trächtige Wistar Ratten in zwei Gruppen geteilt, die beide eine isokalorische Diät bekamen, deren Fettanteil 15 % von frischem (unbenutztem) (P1) bzw. fritiertem (benutztem) (P2) Olivenöl stammte mit nicht trächtigen Ratten verglichen. Die Gravidität erhöhte (p〈0,01) die Futteraufnahme, das Körpergewicht, die Gewichtszunahmen und die Futterverwertung. Die Ölqualität beeinflußte dagegen diese Parameter nicht. Während der Gravidität stiegen die Serumwerte der Triglyceride (TG) (p〈0,01) und des Cholesterins (TC) (p〈0,05) an, während die der Phosphatide (PH) sanken (p〈0,01). Ein signifikanter Effekt der Ölqualität und eine Wechselwirkung zwischen Gravidität und Öl wurde für TG und PH festgestellt. Das Gewicht und der Fettgehalt der Leber der trächtigen Ratten stiegen signifikant an (p.〈0.05), Leber TC, TG und PH stiegen während der Gravidität (ungefähr um das 3-fache der Ausgangswerte), aber es traten keine signifikanten Unterschiede zwischen der Aufnahme von benutztem und nicht benutztem Öl (P2 vs P1) auf. Die Ergebnisse zeigen, daß die Aufnahme von leicht verdorbenem Olivenöl als alleinige Fettquelle der Nahrung keine besonderen Folgen für die Gravidität hat, was die Gewichtszunahme der Mütter und der Feten, die Lipämie und die Zusammensetzung des Leberfetts betrifft.
    Notes: Summary The effect of the consumption of unused olive oil (polar content, 2 %; oleic acid, 78.9 mg/100 mg oil, and linoleic acid 7 mg/100 mg oil) and olive oil used discontinuously for frying potatoes 15 times (polar content, 9 %; oleic acid, 75.8 mg/100 mg oil and linoleic acid 6.2 mg/100 mg oil) was studied in pregnant rats with the aim of better understanding the relationship between the consumption of fat used in frying and lipid metabolism during periods of intense anabolism. Trials were performed in pregnant Wistar rats, divided into 2 groups and fed isocaloric diets in which the fat content (15 % wt/wt) consisted of unused olive oil (P1) or oil previously used for frying (P2), and the results were compared with those of nonpregnant rats fed unused olive oil (NP1) and olive oil used for frying (NP2). Pregnancy increased (p〈0.01) food intake, body weight, weight gain, and food efficiency ratio (P2 vs NP2 and P1 vs NP1, respectively), but the treatment of oil included in the diets did not alter these parameters. Gestation significantly increased the serum triglyceride (TG) (p〈0.01) and total cholesterol (TC) (p〈0.05) concentrations and diminished that of phospholipids (PH) (p〈0.01). A significant effect of the type of oil consumed and a pregnancy x oil interaction on Tg and PH levels was observed. The weight of the liver and its fat content increased significantly (p〈0.05) as a result of pregnancy. Liver TC, TG, and PH increased (approximately 3 times the original values) during gestation, but no significant differences due to the intake of used or unused oil (P2 vs P1) were observed. The results indicate that the consumption of moderately altered olive oil, as the sole source of fat, does not alter the effect of pregnancy on the mothers' weight gain, lipaemia, and hepatic fat composition to any important degree.
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  • 17
    ISSN: 1436-6215
    Keywords: Body composition ; fat ; growth ; somatotropin ; pig ; Körperzusammensetzung ; Fett ; Wachstum ; Somatotropin -Schwein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Zusammenfassung An 78 männlichen Kastraten und weiblichen Schweinen der Kreuzung Pietrain x (Deutsches Edelschwein x Deutsche Landrasse) wurden Untersuchungen zum Einfluß einer porcinen Somatotropin (pST)-Behandlung während des Wachstums auf die chemische Körperzusammensetzung der Tiere, das Adipozyten-Wachstum und das Fettsäure-Profil des Rückenspecks untersucht. Die intramuskulären Injektionen (1 oder 3 mg pST) wurden täglich ab durchschnittlich 65 kg Lebendgewicht bis zum Schlachten verabreicht. Nach der pST-Behandlung wurden bei den Kastraten in allen untersuchten Merkmalen signifikante Unterschiede festgestellt, während sich bei den weiblichen Tieren nur leichte Reaktionen zeigten. Das pST verursachte eine Zunahme des Wasserund Proteingehaltes bei gleichzeitiger Reduzierung des Lipidgehaltes besonders in den fettreichen Körperteilen der Tiere. Im Rückenspeck war der Gehalt an ungesättigten Fettsäuren erhöht und der Fettzelldurchmesser erniedrigt.
    Notes: Summary Seventy eight growing-finishing pigs (male castrates and females) of the cross-breed Pietrain x (Large White x German Landrace) were used to investigate the effects of pST treatment on the chemical composition of the body, the growth of adipocytes, and the fatty acid profile of the backfat. Intramuscular injections (1 or 3 mg pST) were administered daily from an average weight of 65 kg up to slaughter. After pST treatment significant changes in all studied characteristics were observed in barrows, whereas the females exhibited very small responses. The pST caused an increase of water and protein contents and a simultaneous decrease of lipid content especially in body parts rich in fat. Furthermore, the proportion of unsaturated fatty acids increased and the fat cell diameter decreased in the backfat.
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  • 18
    Electronic Resource
    Electronic Resource
    Springer
    Journal of productivity analysis 8 (1997), S. 293-310 
    ISSN: 1573-0441
    Keywords: growth ; USagriculture ; externalities ; spill-overs ; public R and D
    Source: Springer Online Journal Archives 1860-2000
    Topics: Economics
    Notes: Abstract Growth in U.S. agriculture is linked to the non-farm economy through domestic terms of trade and factor market adjustments. With almost stable input growth, the relatively large contributions from growth in Total Factor Productivity (TFP) are passed on to intermediate and final consumers in the form of declining real prices for primary farm products. The resulting net growth in the real value of farm output (GDP) is relatively low (0.25% per annum). The decomposition of TFP suggests that public agricultural stock of knowledge and infrastructure are “robustly” associated with TFP growth, while spill-overs from private agricultural and economy wide research and development (R and D) are positive but, relatively small.
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  • 19
    Electronic Resource
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    Springer
    Journal of insect conservation 1 (1997), S. 43-62 
    ISSN: 1572-9753
    Keywords: Lepidoptera ; biogeography ; endemism ; conservation ; butterflies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Nature of Science, Research, Systems of Higher Education, Museum Science
    Notes: Abstract The addition of species to the European butterfly list since 1983 has resulted in a number of highly significant changes. Most important are the increases in the number and proportion of endemics and of rare species, and a regional excess of species and endemics for southern Europe compared to northern Europe. There is also a surplus of Lycaenidae and Satyridae compared to other families, and an increase in species per genus associated with the reduction in genera. These additions raise two issues. First, the potential conservation load for European butterflies is inflated at species level. This is especially the case for southern Europe, which has disproportionate increases in rare and endemic species, more particularly if rarity and endemism are found to equate with threat of extinction. Second, the inflation in rarity and endemism suggests that there is a trend to promote ever more local populations (races, subspecies) to species. The taxonomic status of species being added to the list, a quarter of which are regarded as doubtful, is increasingly difficult to determine. Consequently, there is a danger that this may call into question the validity and objectivity of taxonomic practices, and of databases dependent on them, used by conservation. Revision of higher and lower butterfly taxa is urgently required.
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  • 20
    Electronic Resource
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    Springer
    Journal of insect conservation 1 (1997), S. 99-111 
    ISSN: 1572-9753
    Keywords: Maculinea ; conservation ; genetic markers ; population structure.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Nature of Science, Research, Systems of Higher Education, Museum Science
    Notes: Abstract We have investigated the genetic population structure within and the genetic differentiation between local populations of the large blue butterfly Maculinea alcon throughout the Jutland peninsula. Samples were collected as eggs on foodplants (Gentiana pneumonanthe), and reared to 4th instar caterpillars in the laboratory. A significant excess of homozygotes was found for all the investigated allozyme loci in most of the populations. A North-South cline was observed for the allele frequencies at some of the loci and for several linkage groups. Because some of the allele frequency clines were parallel to clines in adult morphological variation, we interpret our results as evidence for the co-existence of at least two gene pools within the Danish Maculinea alcon populations. Multilocus electrophoretic data revealed highly positive but variable FST values, which under this scenario would reflect varying frequencies of the Maculinea gene pools across the local populations. The significantly positive FIS values indicate that these gene pools are at least partly reproductively isolated (Wahlund effect). The co-occurrence of several Maculinea alcon gene pools on many local sites in Jutland is of great importance for conservation of the fragmented Maculinea populations. Our results show that there is probably more Maculinea biodiversity to conserve than was previously thought, and suggests that extant populations are more fragmented and vulnerable than counts of flying adults or eggs on foodplants indicate.
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  • 21
    Electronic Resource
    Electronic Resource
    Springer
    Journal of insect conservation 1 (1997), S. 159-166 
    ISSN: 1572-9753
    Keywords: butterfly ; Eurodryas aurinia ; conservation ; Wales ; metapopulation.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Nature of Science, Research, Systems of Higher Education, Museum Science
    Notes: Abstract We surveyed populations of Eurodryas aurinia (a butterfly listed as ‘Threatened in Europe’) in Glamorgan (South Wales, UK). The survey may provide a model for similar work, which is urgently needed throughout the species' European range. For each colony, we established population size, vegetation types, and current management regimes. Populations were assessed using larval surveys, a method which has several advantages over conventional adult surveys. With approximately 35 local populations, Glamorgan is among the most important areas for E. aurinia in the UK, and is of importance in a European context. However, 15 local populations were under immediate threat from unfavourable management or industrial developments, and only seven populations were on Sites of Special Scientific Interest. Colonies showed a clustered pattern, and varied greatly in size: 50% supported 20 or fewer larval webs. Many of the small populations may be temporary offshoots of larger, more permanent populations nearby. The largest local populations occupied Molinia caerulea - Cirsium dissectum fen meadow habitats (National Vegetation Community M24), which were unmanaged, grazed by cattle, horses or ponies, or subject to periodic burning. Detailed local surveys such as this one, which assess the relative sizes of populations and the impact of current management practices, may be the best way to plan future conservation measures for E. aurinia.
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  • 22
    ISSN: 1432-1939
    Keywords: Key words Agrostis canina ; CO2 vents ; photosynthesis ; lignification ; growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The aim of this study was to characterise growth and photosynthetic capacity in plants adapted to long-term contrasting atmospheric CO2 concentrations (C a). Seeds of Agrostis canina L. ssp. monteluccii were collected from a natural CO2 transect in central-western Italy and plants grown in controlled environment chambers at both ambient and elevated CO2 (350 and 700 μmol mol−1) in nutrient-rich soil. Seasonal mean C a at the source of the plant material ranged from 610 to 451 μmol CO2 mol−1, derived from C4 leaf stable carbon isotope discrimination (δ13C). Under chamber conditions, CO2 enrichment stimulated the growth of all populations. However, plants originating from elevated C a exhibited higher initial relative growth rates (RGRs) irrespective of chamber CO2 concentrations and a positive relationship was found between RGR and C a at the seed source. Seed weight was positively correlated with C a, but differences in seed weight were found to explain no more than 34% of the variation in RGRs at elevated CO2. Longer-term experiments (over 98 days) on two populations originating from the extremes of the transect (451 and 610 μmol CO2 mol−1) indicated that differences in growth between populations were maintained when plants were grown at both 350 and 700 μmol CO2 mol−1. Analysis of leaf material revealed an increase in the cell wall fraction (CWF) in plants grown at elevated CO2, with plants originating from high C a exhibiting constitutively lower levels but a variable response in terms of the degree of lignification. In vivo gas exchange measurements revealed no significant differences in light and CO2 saturated rates of photosynthesis and carboxylation efficiency between populations or with CO2 treatment. Moreover, SDS-PAGE/ LISA quantification of leaf ribulose bisphosphate carboxylase/oxygenase (Rubisco) showed no difference in Rubisco content between populations or CO2 treatments. These findings suggest that long-term adaptation to growth at elevated CO2 may be associated with a potential for increased growth, but this does not appear to be linked with differences in the intrinsic capacity for photosynthesis.
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  • 23
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    Journal of thermal analysis and calorimetry 49 (1997), S. 1571-1584 
    ISSN: 1572-8943
    Keywords: conservation ; environmental research for art
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The basic concept of this project is to identify and then use the changes which occur in the chemical and physical properties of traditional paint media both to indicate and integrate the effects of environmental conditions on paintings. To achieve this aim, test paint films are being prepared in accordance with traditional artists' recipes. Changes in material properties are monitored using a combination of non-invasive spectroscopy (Bacci), microsensors, thermoanalytical techniques (Odlyha), and microscale analytical mass spectrometry for molecular structure analysis (Boon). The test strips are calibrated by exposure to controlled environments (light, temperature, relative humidity and noxious gases) and alterations in their properties are quantified. This provides information on the nature and rate of change at the molecular level and a data base for evaluating the molecular monitoring strips after their exposure in the field. Field sites have been selected and include various locations in the Tate Gallery (UK), Sandham Chapel (Burghclere, UK), the Uffizi Gallery (It) and the Rijksmuseum (NL). Environmental conditions of some of these locations are being evaluated at present using the glass sensors described in project EV5VCT92 0144. Small piezoelectric quartz crystal humidity sensors will be installed to determine localised variations in relative humidity and temperature on [1] Stanley Spencer paintings in Sandham Chapel and [2] Giotto's “Madonna di Ognissanti” in the Uffizi Gallery. In addition novel coatings using picture varnishes are being applied to similar piezoelectric quartz crystal sensors to evaluate the effects of environmental impact on the chemistry of varnishes on paintings. Data are also being collected on the nature of chemical and physical changes in varnishes and paint media in actual paintings at the molecular level.
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  • 24
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    Journal of thermal analysis and calorimetry 50 (1997), S. 191-202 
    ISSN: 1572-8943
    Keywords: conservation ; deacidification procedures ; TMA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The use of non-aqueous deacidification procedures as a preventive conservation measure to assist in retarding the deterioration of painting canvases has been suggested by the Conservation Department of the Tate Gallery [1]. The reverse sides of paintings are treated with commercially available MMC solution (methoxy magnesium methyl carbonate). The aim of this paper is to describe how dynamic mechanical thermal analysis can be used to evaluate the effects of this treatment. Measurements are described on modern commercially primed canvas samples [2] which show that the MMC treatment does cause an increase in the modulus or stiffness of the primed canvas materials but that the effect on theT g is minimal. The response of the treated materials to variations in relative humidity has also been studied and indications are that the response of treated canvases to variations in relative humidity differs from those of the untreated canvases.
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  • 25
    ISSN: 1573-4919
    Keywords: calreticulin ; gene expression ; steroid receptor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Calreticulin is a ubiquitously expressed Ca2+ binding protein of the endoplasmic reticulum which inhibits DNA binding and transcriptional activation by steroid hormone receptors. In this study the effects of calreticulin on tyrosine aminotransferase (TAT) gene expression in cultured McA–RH7777 hepatocytes was investigated. McA–RH7777 cells were stably transfected with calreticulin expression vector to generate cells overexpressing the protein. The transcriptional activity of the TAT gene, which is glucocorticoid–sensitive and cAMP–dependent, was investigated in the mock transfected McA–RH7777 and in cells overexpressing calreticulin (designated McA–11 and McA–17). In the presence of dexamethasone or the cAMP analog (CTP–cAMP) expression of the TAT gene was induced in mock transfected McA–RH7777 cells by approximately 4.5 and 5 fold, respectively. In McA–11 and McA–17 cells, overexpressing calreticulin, glucocorticoi ever, the CTP–cAMP–dependent expression of the TAT gene was not affected. The ability of calreticulin to inhibit glucocorticoid–sensitive TAT gene expression but not the cAMP–dependent expression of the gene suggests that the protein affects specifically the action of transcription pathways involving steroid receptors or transcription factors containing KxFF(K/R)R–like motifs. Calreticulin may play an important role in the regulation of glucocorticoid–sensitive pathway of expression of the hepatocytes specific genes during development.
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  • 26
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; diabetic state ; ethanol ; liver injury
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The alteration in calcium-binding protein regucalcin in the liver and serum of rats with streptozotocin (STZ)-diabetic state or ethanol ingestion was investigated. STZ (6.0 mg/100 g body weight) was subcutaneously administered in rats, and 1 or 3 weeks later they were sacrificed by bleeding. Liver regucalcin mRNA levels were not clearly altered by the diabetic state, as evidenced by Northern blotting using regucalcin cDNA (0.9 kb of open reading frame). Based on enzyme-linked immunoadsorbent assay (ELISA) with rabbit-anti-regucalcin IgG, hepatic regucalcin concentration was decreased about 50% of control levels by STZ treatment. However, serum regucalcin concentration was not significantly altered by STZ treatment. Meanwhile, when rats ingested ethanol (10 and 30%) in the drinking water for 2 weeks, liver regucalcin mRNA levels were clearly increased, although hepatic regucalcin concentration was significantly decreased. Serum regucalcin concentration was not appreciably altered. Serum transaminases (GOT and GPT) activities were significantly increased at 1 or 3 weeks after STZ administration in rats, while their activities were not altered by ethanol ingestion. The present study demonstrates that hepatic regucalcin concentration is decreased independent of mRNA expression in the STZ-diabetes and during ethanol ingestion in rats.
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  • 27
    ISSN: 1573-4919
    Keywords: NADH oxidase ; plasma membranes ; growth ; retinol ; retinoids ; HeLa cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Several retinoids, both natural and synthetic, were evaluated for their ability to modulate NADH oxidase activity of plasma membranes of cultured HeLa cells and the growth of HeLa cells in culture. Both NADH oxidase activity and the growth of cells were inhibited by the naturally-occurring retinoids all trans-retinoic acid (tretinoin) and retinol as well as by the synthetic retinoids, trans-acitretin, 13-cis-acitretin, etretinate and arotonoid ethylester (Ro 13-6298). For all retinoids tested, inhibition of NADH oxidase activity and inhibition of growth were correlated closely. With tretinoin, etretinate and arotonoid ethylester, NADH oxidase activity and cell growth were inhibited in parallel in proportion to the logarithm of retinoid concentration over the range of concentrations 10-8 to 10-5 M. Approximately 70% inhibition of both NADH oxidase activity and growth was reached at 10 µM. With retinol, trans-acitretin and 13-cis-acitretin, inhibition of NADH oxidase activity and growth also were correlated but maximum inhibition of both was about 40% at 10 µM. The possibility is suggested that inhibition of the plasma membrane NADH oxidase activity by retinoids may be related to their mechanism of inhibition of growth of HeLa cells in culture. (Mol Cell Biochem 166: 101-109, 1997)
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  • 28
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    Molecular and cellular biochemistry 167 (1997), S. 169-177 
    ISSN: 1573-4919
    Keywords: tamoxifen ; interferon ; gene expression ; breast cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The molecular basis for the enhanced growth inhibition of MCF-7 human breast cancer xenografts by a combination of human interferon-β (IFN-β) and tamoxifen was investigated. Treatment of MCF-7, MDA-MB-231, and BT-20 cells with the combination of IFN-β and tamoxifen resulted in enhanced antiproliferative effects in vitro. Treatment with the combination of IFN-β and tamoxifen enhanced the expression of several IFN-β-inducible genes in human breast carcinoma cell lines relative to levels induced by IFN-β alone. Tamoxifen alone did not induce transcription of IFN-stimulated genes (ISGs). Augmentation of ISG expression by the combination of IFN-β and tamoxifen was noted in breast tumor cell lines irrespective of their functional estrogen receptor (ER) status or their dependence on estradiol for growth, suggesting that upregulation of ISGs was independent of ER status. Enhancement of IFN-stimulated gene expression by tamoxifen occurred at the transcripti onal level. Expression of transfected reporter genes under the control of IFN-α/β regulated promoters was also enhanced in IFN-β and tamoxifen-treated cells. Similarly, transcriptional induction of chimeric reporter plasmids driven by an IFN-γ inducible promoter (GAS; IFN-γ activated site) was also enhanced by the combination of IFN-γ and tamoxifen. In tamoxifen treated cells, IFN-β and IFN-γ readily activated transcription factors ISGF-3 and GAF, respectively. Therefore, augmentation of ISG expression by tamoxifen is an early event in the antitumoral activity of this drug combination. (Mol Cell Biochem 167: 169-177, 1997)
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  • 29
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    Molecular and cellular biochemistry 172 (1997), S. 47-57 
    ISSN: 1573-4919
    Keywords: smooth muscle ; gene transfer ; DNA ; RNA ; ribozyme ; liposome ; lipoxygenase ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Chemically synthesized hammerhead-type ribozymes targeted against the porcine leukocyte-type 12-lipoxygenase (LO) have been developed and studied. One chimeric ribozyme consists of DNA in the non-enzymatic portions, and RNA in the enzymatic core as well as two phosphorothioate internucleotide linkages at 3′ terminus. The second ribozyme consists of ribonucleotide sequences generated by in vitro transcription. In this chapter we describe methodologies to first analyze the ribozyme catalytic activity in vitro by studying cleavage of target RNA in vitro. The subsequent sections will describe how to target the catalytic ribozyme and deliver it to porcine vascular smooth muscle cells (PVSMC) by a liposome-mediated method. Finally ways to evaluate its activity to inhibit expression of the 12-LO mRNA will be presented. These results demonstrate the feasibility of using ribozymes as novel candidates for therapeutic agents to block specific gene expression in vascular cells.
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  • 30
    ISSN: 1573-4919
    Keywords: heart ; DNA ; library ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The availability of high quality cDNA libraries is often crucial to the successful identification and characterization of genes. The concepts and potential pitfalls of constructing cDNA libraries are presented. Various applications requiring high quality cDNA libraries are outlined, including large-scale single pass sequencing of cDNA clones to generate expressed sequence tags (ESTs) and differential screening of cDNA libraries. The usefulness of combining such approaches for the discovery of novel disease-related and cardiovascular-based ESTs (CVBest) is discussed.
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  • 31
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    Molecular and cellular biochemistry 173 (1997), S. 59-69 
    ISSN: 1573-4919
    Keywords: hydrogen peroxide ; oxidative stress ; gene expression ; lens epithelial cells ; N-acetylcysteine ; pyrrolidine dithiocarbamate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The involvement of H2O2 in cataract development has been established inboth human patients and animal models. At the molecular level H2O2 has beenobserved to cause damage to DNA, protein and lipid. To explore the oxidativestress response of the lens system at the gene expression level, we haveexamined the effects of H2O2 on the mRNA change of the proto-oncogenes,c-jun, c-fos and c-myc in a rabbit lens cell line, N/N1003A. H2O2 treatmentof the rabbit lens epithelial cells for 60 min induces quick up-regulationof both c-jun and c-fos mRNAs. The maximal induction is 38 fold for c-jun at150 µM and 72 fold for c-fos at 250 µM H2O2. Treatment ofN/N1003A cells with 50-250 µM H2O2 for 60 min leads to a 2-5 foldincrease of the c-myc mRNA level. H2O2 also induces an up-regulation intransactivity of the activating protein-1 (AP-1) as shown with a reportergene driven by a prolactin gene promoter with 4 copies of AP-1 binding sitesinserted in the upstream of the promoter. Maximal induction occurs with 150µM H2O2. In the same system, the antioxidants, N-acetyl-cysteine (NAC)and pyrrolidine dithiocarbamate (PDTC) at concentrations shown toup-regulate the mRNAs of both c-jun and c-fos, also enhance thetransactivity of AP-1. NAC and PDTC have different effects in modulating theinduction of AP-1 activity by H2O2 and TPA. These results reveal thatoxidative stress regulates expression of various regulatory genes in lenssystems, which likely affects cell proliferation, differentiation andviability and thus affect normal lens functions.
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  • 32
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; cDNA cloning ; gene expression ; mouse liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The molecular cloning of the cDNA coding for a Ca2+-binding proteinregucalcin and its mRNA expression in mouse liver were investigated. ThecDNA clone encoding a regucalcin was isolated from a mouse liver cDNAlibrary and sequenced. Analysis of the sequence of the cloned cDNA showedthat the cDNA encoded the complete amino acid sequence of the mouseregucalcin molecule; the cDNA had an open reading frame of 897 bp. Mouseregucalcin was composed of 299 amino acid residues, and its molecular weightwas estimated to be 33,406 Da. The amino acid sequence of mouse regucalcinhad 94% homology, as compared with that of rat regucalcin. Northernblot analysis with the mouse liver cDNA probe revealed that mouse regucalcinmRNA was mainly present in the liver but only slightly in the kidney with asize of 1.8 kb. Hepatic regucalcin mRNA level of male mouse was higher thanthat of female mouse. A single intraperitoneal administration of calciumchloride (5, 15, and 30 mg Ca2+/100 g body weight) to mice induced aremarkable increase in regucalcin mRNA in the liver; the increase inregucalcin mRNA levels at 30 min after calcium administration wasdose-dependent. The present results demonstrate that regucalcin mRNA in miceis uniquely expressed in the liver, and that its expression is stimulated bycalcium administration.
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  • 33
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    Molecular and cellular biochemistry 176 (1997), S. 273-279 
    ISSN: 1573-4919
    Keywords: cardiac hypertrophy ; myosin heavy chain ; gene expression ; adrenergic system
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Growth of the heart in hypertrophy is accompanied by changes in the phenotypic expression of cardiac genes. To explore the molecular basis of cardiac hypertrophy, we have analyzed the regulation of myosin heavy chain gene (MHC) expression. In one set of experiments, pressure overload on the rat heart was produced by constriction of the abdominal aorta. Changes in the α and β-MHC mRNA were then studied in overloaded hearts and following load removal. Pressure overload resulted in down-regulation of the α-MHC with corresponding up-regulation of the steady state level of β-MHC mRNA. Load removal (debanding) resulted in regression of cardiac hypertrophy and a rapid return of α-MHC mRNA to normal values. In contrast, the recovery in β-MHC mRNA was much slower to the extent that it remained substantially elevated compared to respective sham controls even after 7 weeks of post-debanding. These results suggest that putative load-related signals independently regulate two genes. Several lines of evidence indicate that adrenergic nervous system plays an important role in the induction and maintenance of cardiac hypertrophy and in the redistribution of myosin isoforms. We have analyzed the effect of cAMP inducing agents on the regulation of a-MHC gene in primary cultures of the fetal (18 day) rat cardiac myocyte. Inclusion of 8 Br-cAMP in the culture media increased the expression of α-MHC promoter/reporter construct comprising of 2.9 kb upstream sequence of the α-MHC gene. Several deletion mutations in the α- MHC gene promoter defined the cAMP responsive boundaries to be a 32 bp region comprising of -71 to -40 bp sequences. Deletion of this region resulted in loss of cAMP response as well as in basal expression of α-MHC promoter/reporter construct. These data suggest a role of β-adrenergic pathway in the modulation of α-MHC gene expression.
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  • 34
    ISSN: 1573-4927
    Keywords: glycophorins ; gorilla ; evolution ; gene family ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Homologues of MN blood group antigens, encoded by members of the glycophorin A (GPA) gene family, are expressed in man, anthropoid apes, and some species of Old World monkeys. Previous studies had shown that a three-gene framework, most closely related to that in man, is present in the chimpanzee. Here we report the genomic structure, transcript map, and protein expression of the GYPA locus in gorillas. Compared to the corresponding human and chimpanzee homologues, gorilla GPA, GPB, and GPB/E genes each showed a high degree of sequence identity, with the same exon-intron organization. However, the expression of exons III, IV, or V encoding the extracellular or membrane domains of homologous glycophorins varied among the three species. Gorilla GPA and GPB/E genes were unique in that the former occurred in two allelic forms with or without the expression of exon III, whereas the latter contained one (ψ exon III) instead of two silenced exons (ψ exons III and IV). Differences from human but not chimpanzee GPA also included the presence of a hybrid M/N epitope and the absence of the sequon for N-glycosylation. Owing to the retention of a functional exon III, gorilla GPB was more similar to chimpanzee GPB than human GPB. A transspecies allele was identified in the gorilla that gave rise to the Henshaw (He)-like antigen similar to that found in man. These results provide further insight into the model for evolution of the GPA gene family, indicating that the mechanisms underlying inter- and intraspecific polymorphism of glycophorins could predate the divergence of gorillas as the consequence of gene duplication and diversification.
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  • 35
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    Molecular and cellular biochemistry 172 (1997), S. 37-46 
    ISSN: 1573-4919
    Keywords: gene transfer ; gene expression ; adenovirus ; blood vessel
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Adenovirus-mediated gene transfer is a promising method for studies of vascular biology and potentially for gene therapy. Intravascular approaches for gene transfer to blood vessels in vivo generally require interruption of blood flow and have several limitations. We have used two alternative approaches for gene transfer to blood vessels in vivo using perivascular application of vectors. First, replication-deficient adenovirus expressing nuclear-targeted bacterial b-galactosidase was injected into cerebrospinal fluid via the cisterna magna of rats. Leptomeningeal cells over the major arteries were efficiently transfected, and adventitial cells of large vessels and smooth muscle cells of small vessels were occasionally stained. When viral suspension was injected with the rat in a lateral position, the reporter gene was expressed extensively on the ipsilateral surface of the brain. Thus, adenovirus injected into cerebrospinal fluid provides gene transfer in vivo to cerebral blood vessels and, with greater efficiency, to perivascular tissue. Furthermore, positioning of the head may ‘target’ specific regions of the brain. Second, vascular gene delivery was accomplished by perivascular injection of virus in peripheral vessels. Injection of the adenoviral vector within the periarterial sheath of monkeys resulted in gene transfer to the vessel wall that was substantial in magnitude although limited to cells in the adventitia. Approximately20% of adventitial cells expressed the transgene, with no gene transfer to cells in the intima or media. These approaches may provide alternative approaches for gene transfer to blood vessels, and may be useful for studies of vascular biology and perhaps vascular gene therapy.
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  • 36
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    Molecular and cellular biochemistry 172 (1997), S. 111-120 
    ISSN: 1573-4919
    Keywords: differential display ; cardiac development ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract An estimated 15,000 different mRNA species are expressed in a typical mammalian cell. The differential expression of mRNAs in both a temporal and cell-specific manner determines the fate of the cell and creates the organism. Analysis of this differential gene expression has become a central aim of many laboratories attempting to understand the mechanisms underlying various biological processes. Currently, we are using a technique called differential display to analyze the differential expression of genes in cardiomyocytes. Differential display is a rapid and powerful technique that was introduced by Liang and Pardee in 1992. Since that time, it has been successfully applied by several groups, and it is quickly becoming a standard method for studying differential gene expression. Here, we present a detailed article discussing the differential display methodology and how we have utilized it to identify potential genes involved in cardiomyocyte proliferation. Furthermore, we have provided a list of materials and supplied examples of data obtained, in an effort to allow the reader to perform the technique with success in their own laboratory.
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  • 37
    ISSN: 1573-4919
    Keywords: plasminogen activators ; plasminogen activator inhibitors ; gene expression ; left ventricular hypertrophy ; pressure overload
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract In the early stages of left ventricular hypertrophy (LVH) acute adaptive changes occur in the coronary vasculature as it remodels. Plasminogen activators (PAs) and inhibitors (PAIs) have the potential effects of proteolytic degradation that is relevant to tissue remodeling and angiogenesis. Our study focused on the possible roles of PAI-1, PAI-2, uPA and tPA in myocyte hypertrophy and angiogenesis in the early and late stages of pressure overload induced left ventricular hypertrophy (LVH). We divided seventeen adult swine, weighing 24.2 ± 6.5 kg, into four groups: control, sham-operated, early LVH and late heart failure LVH group. At surgery we placed a fixed constrictor on the ascending aorta immediately above the aortic valve. This increased LV systolic pressure from 133 ± 15 to 193 ± 24 mm Hg after the surgery. We subdivided the early group into groups of 3 animals each that we euthanized at 8, 24 and 72 h after operation and obtained heart samples for analysis. In the late heart failure group individual animals were euthanized at 55, 59, 62 and 72 days after the detection of congestive heart failure. We also obtained tissue samples from the control and sham-operated swine. Sections for histologic analysis were fixed in 10% buffered formalin. We isolated RNA, size fractionated it using 1% formaldehyde-agarose gel electrophoresis and then did Northern blots. The mRNAs from both PAI-1 and PAI-2 showed a remarkable increase at 8 and 24 h after acute aortic constriction and returned to control by 72 h. Regional differences showed that most of the increases were in the endocardium. Three animals in the late heart failure LVH group were determined to be in congestive heart failure at about 2 months after the onset of aortic constriction. In these animals PAI-1 and PAI-2 were increased in both the left and right ventricles but remained low in an animal of the same elevation in aortic pressure seen by the LV who did not have congestive failure. These data suggest that PA and PAI gene expressions change before morphologic changes occur in the early stages of developing LVH. Also at the time of onset of congestive heart failure this increased expression reappears. PAs and PA inhibitors mRNA levels vary in the different regions of the heart reflecting changing wall stresses. Thus, the PAs and PA inhibitors may play an important role in angiogenesis that occurs during the early stages of LVH. The increased expression in the late stage of LVH may reflect further changes in wall stresses since these animals also showed overt clinical signs of heart failure.
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  • 38
    ISSN: 1573-4919
    Keywords: estrogen ; apolipoprotein ; gene expression ; mice ; atherosclerosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Estrogen protects against developing premature coronary artery disease.However, the mechanism of protective effects of estrogen still remainspoorly understood. One mechanism by which estrogen can have protectiveeffects apppears to be through modulation of plasma lipoproteins. We showedthat the mouse can be used as animal model to study estrogen-mediatedsynthesis and secretion of lipoproteins since, unlike the rat, the mousedoes not up-regulate LDL receptors (Srivastava et al. [4]). Since inbredstrains of mice differ in their genetic background and show differingresponsiveness to dietary lipids, we examined how various inbred strains ofmice respond to estradiol administration, and whether some mouse strainsshow responses similar to rats. 17b-estradiol was administered to male micefrom 15 different inbred strains, and the changes in plasma levels oflipids, apoB, apoAI, and apoE were examined. Total cholesterol decreased inall but one strain, apoAI levels decreased in all but 3 strains while apoBlevels and apoB/apoAI ratios increased in all but 2 strains, suggesting thatin contrast to rats, the apoB-containing lipoproteins increased relative toHDL in all strains of mice examined. Basal and estradiol-induced changes intotal cholesterol were significantly correlated with changes in apoAI, butnot apoB, reflecting the predominance of HDL over other lipoproteins inmouse plasma. The effects of estrogen on plasma apoE levels varied amongvarious inbred strains of mice tested. Plasma apoE levels increased in sevenstrains treated with estrogen, and remained unchanged in the rest. Toexamine whether changes of plasma apoproteins are associated with thechanges in the respective hepatic mRNA levels, apoAI, B and E mRNA werequantified by RNase protection assay. Hepatic apoE mRNA did not showcorrelation with either basal or post treatment plasma apoE levels in any ofthe strains. Similarly, most of the mouse strains did not show correlationof plasma apoAI and apoB levels with the corresponding hepatic mRNA levels.These results suggest that estrogen regulates plasma lipoproteinconcentrations primarily by posttranscriptional mechansims, and there werestrain-related differences in the estrogen-mediated regulation oflipoprotein metabolism.
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  • 39
    ISSN: 1573-4919
    Keywords: regucalcin ; Ca2+-binding protein ; insulin ; gene expression ; HepG2 cells ; transformed cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The expression of hepatic Ca2+-binding protein regucalcin in the cloned human hepatoma cells (HepG2) was investigated. The change in regucalcin mRNA levels was analyzed by Northern blotting using rat liver regucalcin complementary DNA (0.9 kb of open reading frame). Regucalcin mRNA was expressed in HepG2 cells, although the mRNA was markedly expressed in normal rat liver. Moreover, regucalcin protein in HepG2 cells was detected by Western blot analysis using a polyclonal rabbit anti-regucalcin antibody. Regucalcin mRNA expression in HepG2 cells was clearly stimulated by the culture with insulin (10-8 M) of the effective concentration. Regucalcin protein in HepG2 cells was also increased by the treatment of insulin (10-8 M). The present results demonstrate that regucalcin is expressed in the transformed HepG2 cells, and that the expression is stimulated by insulin.
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  • 40
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    Molecular and cellular biochemistry 177 (1997), S. 1-6 
    ISSN: 1573-4919
    Keywords: gene expression ; mRNA secondary structure ; single tube RT-PCR ; TNF receptor I
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The secondary structure of human tumor necrosis factor receptor I (TNFR-I) mRNA based on its lowest folding energy was predicted. Three combinations of primers selected from open-regions and four combinations of primers from closed-regions of TNFR-I mRNA structure were employed for single-tube reverse transcription-polymerase chain reaction (RT-PCR) for the determination of TNFR-I gene expression in U937 cell. All the primers were designed with the same criteria. However, the different primers generated distinct quantities of RT-PCR products from the same concentration of TNFR-I mRNA, implying that the determination of gene expression by RT-PCR was affected by the mRNA secondary structure. In addition, the sensitivity of the open-region RT-PCR was approximately one hundred-fold higher than that in the closed-regions of TNFR-I mRNA. The low efficiency of the closed-region RT-PCR was not correlated with the G/C content of the TNFR-I mRNA structure. These results suggest that consideration of the influence of intrinsic mRNA structure of a gene is essential prior to the determination of gene expression by quantitative RT-PCR, and this open-region strategy of primer design may yield an efficient primer for in vitro amplification of cDNA by RT-PCR.
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  • 41
    ISSN: 1573-4919
    Keywords: cholesteryl ester ; CETP ; Caco-2 ; polymerase chain reaction ; gene expression ; mRNA ; alternative splicing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Cholesteryl ester transfer protein (CETP) is a plasma protein involved in the reverse cholesterol transport and expressed in several human tissues and cell lines. We studied CETP expression in Caco-2 cell line, a model of the human enterocyte epithelium. By reverse-transcriptase polymerase chain reaction, we could demonstrate that in basal condition Caco-2 cells have a low rate of expression of active CETP mRNA. Furthermore, we found that even in this cell line CETP mRNA alternative splicing occurs with deletion of exon 9 sequence. Densitometric analysis of the in vitro amplified fragments showed that under basal conditions about 60% of reverse transcribed CETP cDNA corresponds to exon 9-deleted transcripts. After challenge with 50 µM sodium oleate, there is a ∼2 fold increase in the transcription rate of the full-length CETP cDNA, as measured by competitive PCR, which is accompanied to an increased activity measured in the cell-conditioned medium. On the contrary, no significant change is seen in the amount of exon 9-deleted cDNA. Consequently, an inversion in the ratio of full-length and exon 9-deleted CETP cDNA is evident, suggesting that sodium oleate selectively enhances the expression of full-length CETP mRNA.
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  • 42
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    Molecular biology reports 24 (1997), S. 221-230 
    ISSN: 1573-4978
    Keywords: gene expression ; ribonucleoprotein ; RNase MRP ; RNase P ; transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report on the expression of mouse RNase MRP RNA in human embryonic kidney 293 cells upon DNA transfection. Stable cell lines were selected by cotransfection with a neo r gene. Transcription of wild-type and deletion mutants of MRP RNA and ribonucleoprotein formation were assessed by RNase protection and immunoprecipitation experiments. Mouse MRP RNA as expressed in 293 cells readily associates with human proteins to form a chimeric Th ribonucleoprotein. 5' truncated MRP RNAs, however, failed to associate with Th antigen(s) and deletion of the 3' sequences of MRP RNA greatly reduced the expression in stable as well as in transient transfectants. Abbreviations: nt(s) – nucleotide(s); RNP – ribonucleoprotein.
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  • 43
    ISSN: 1573-5028
    Keywords: cDNA cloning ; ethylene ; fruit ripening ; gene expression ; melon ; wounding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In vitro translation of mRNAs and polyacrylamide gel electrophoresis of proteins from melons revealed that several mRNAs increased in amount during ripening, indicating the existence of other ripening genes in addition to those cloned previously. To identify ripening-related genes we have screened a ripe melon cDNA library and isolated two novel cDNA clones (MEL2 and MEL7) encoding unidentified proteins. Southern analysis revealed that MEL2 and MEL7 are encoded by low-copy-number genes. The MEL2 cDNA clone is near full-length, corresponds to a 1600 nucleotide mRNA that accumulates during ripening and encodes a predicted protein rich in hydrophobic amino acids. The MEL7 cDNA clone is full-length, corresponds to a mRNA of 0.7 kb which accumulates during early ripening stages and is also present at low levels in other organs of the melon plant. The MEL7 predicted polypeptide is 17 kDa and shows significant homology with the major latex protein from opium-poppy. Wounding and ethylene treatment of unripe melon fruits 20 days after anthesis showed that MEL2 and MEL7 mRNAs are only induced by ethylene.
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  • 44
    ISSN: 1573-5028
    Keywords: gene expression ; heat shock ; oxidative stress ; ozone ; pathogenesis-related protein ; Petroselinum crispum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Parsley (Petroselinum crispum L.) is known to respond to pathogen attack by the synthesis of furanocoumarins and to UV irradiation by the synthesis of flavone glycosides whereas ozone treatment results in the induction of both pathways. A cDNA library from parsley plants was differentially screened using labelled reverse-transcribed poly(A)+ RNA isolated from ozone-treated parsley plants. This resulted in the isolation of 13 independent cDNA clones representing ozone-induced genes and of 11 cDNA clones representing ozone-repressed genes. DNA sequencing of several clones resulted in the identification of pathogenesis-related protein 1-3 (PR1-3), of a new member of PR1 cDNAs (PR1-4) and of a small heat shock protein (sHSP). Northern blot analyses showed a transient induction of the three mRNA species after ozone fumigation. In contrast, heat shock treatment of parsley plants resulted in an increase of sHSP mRNA whereas no increase for transcripts of PR1-3 and PR1-4 could be observed. This is the first characterized sHSP cDNA clone for plants induced by heat shock, as well as by oxidative stress caused by ozone.
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  • 45
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    Plant molecular biology 33 (1997), S. 301-311 
    ISSN: 1573-5028
    Keywords: asparagine ; asparagine synthetase ; cDNA clone ; complementation ; gene expression ; Glycine max
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two cDNA clones (SAS1 and SAS2) encoding different isoforms of asparagine synthetase (AS; EC 6.3.5.4) were isolated. Their DNA sequences were determined and compared. The amino-terminal residues of the predicted SAS1 and SAS2 proteins were identical to those of the glutamine binding domain of AS from pea, asparagus, Arabidopsis and human, suggesting that SAS1 and SAS2 cDNAs encode the glutamine-dependent form of AS. The open reading frames of SAS1 and SAS2 encode a protein of 579 and 581 amino acids with predicted molecular weights of 65 182 and 65 608 Da respectively. Similarity of the deduced amino acid sequences of SAS1 and SAS2 with other known AS sequences were 92% and 93% for pea AS1; 91% and 96% for pea AS2; 88% and 91% for asparagus; 88% and 90.5% for Arabidopsis; 70.5% and 72.5% for E. coli asnB and 61% and 63% for man. A plasmid, pSAS2E, was constructed to express the soybean AS protein in Escherichia coli. Complementation experiments revealed that the soybean AS protein was functional in E. coli. Southern blot analysis indicated that the soybean AS is part of a small gene family. AS transcript was expressed in all tissues examined, but higher levels were seen in stem and root of light-grown tissue and leaves of dark-treated tissue.
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  • 46
    ISSN: 1573-5028
    Keywords: in vivo protein synthesis ; gene expression ; germination ; isocitrate lyase ; isoform ; megagametophyte ; Pinus taeda L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two full-length cDNAs encoding the glyoxysomal enzyme isocitrate lyase(ICL) were isolated from a λZAP cDNA library prepared frommegagametophyte mRNAs extracted from seeds imbibed at 30 °C for 8days. The cDNAs, designated Ptbs ICL 8 and Ptbs ICL 12, have openreading frames of 1740 and 1719 bp, with deduced amino acid sequences of580 and 573 residues, respectively. The predicted amino acid sequencesof Ptbs ICL 8 and Ptbs ICL 12 exhibit a 79% identity with each other,and have a greater than 75% identity with ICLs from variousangiosperm species. The C-termini of Ptbs ICL 8 and Ptbs ICL 12terminate with the tripeptide Ser-Arg-Met and Ala-Arg-Met, respectively,both being conserved variants of the type 1 peroxisomal targetingsignal. RNA blot and slot analysis revealed that Ptbs ICL 8 and PtbsICL 12 mRNAs were present at low levels in the megagametophyte of themature and stratified seeds, and that the level of both transcriptsincreased markedly upon seed germination. Protein blot analysisindicated that the steady-state level of ICL was low in the mature andstratified seed, then increased rapidly upon seed germination, peakingat around 8-10 days after imbibition (DAI). Changes in the level ofICL activity in cell-free extracts was similar to the steady-stateprotein content with the exception that ICL activity was not detected inmegagametophyte extracts of mature or stratified seeds. From 10-12 DAIwhen the megagametophyte tissue senesced, ICL activity decreased rapidlyto near undetectable levels. In contrast, steady-state levels of ICLprotein and mRNA remained relatively constant during megagametophytesenescence. In vivo synthesis of ICL protein was measured to shedlight on these differences. ICL immunoselected from[35S]-methionine labelled proteins indicated that ICL wassynthesized at very low levels during megagametophyte senescence.Together, the results show that loblolly pine ICL gene expression iscomplex. While temporal regulation appears to be primarilytranscriptional, it also involves a number of post-transcriptionalprocesses including at least one translational and/or post-translationalmechanism.
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  • 47
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    Plant molecular biology 34 (1997), S. 345-352 
    ISSN: 1573-5028
    Keywords: heat shock ; pea (Pisum sativum L.) ; gene expression ; pod lignification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A pea pod cDNA library was screened for sequences specific to lignifying tissue. A cDNA clone (pLP19) encoding the C-terminal region of a hsp70 heat shock protein hybridised only to pod mRNA from pea lines where pod lignification occurred. Expression of pLP19 was induced by heat shock in leaves, stems and roots of pea and chickpea plants. Four different poly(A) addition sites were observed in cDNAs derived from the same gene as pLP19. This gene was fully sequenced; unlike most hsp70 genes, it contains no introns. The 5′-flanking sequence contains heat shock elements and other potential regulatory sequences.
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  • 48
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    Plant molecular biology 33 (1997), S. 821-834 
    ISSN: 1573-5028
    Keywords: leaf senescence ; genes ; gene expression ; subtractive hybridisation ; Brassica napus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A subtractive hybridisation technique was developed to clone cDNAs representing genes that showed enhanced expression during leaf senescence in Brassica napus. A number of different genes were identified that, when analysed by northern hybridisation, showed different patterns of expression during leaf development but were all expressed at increased levels during senescence. Sequence analysis of these cDNAs showed that several types of genes were found including two different proteases, glutamine synthetase, ATP sulphurylase, catalase, metallothionein, ferritin and an antifungal protein. The possible roles of these gene products in the senescence process are discussed.
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  • 49
    ISSN: 1573-5028
    Keywords: gene expression ; grass ; Phalaris ; self-incompatibility ; thioredoxin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Self-incompatibility is widespread in the grasses and it is proposed that the grasses share a common incompatibility mechanism that is distinct from those operating in the dicotyledonous species studied in great detail. Where good genetic data are available, all grass species appear to have an incompatibility mechanism controlled by two unlinked loci, S and Z. A putative S gene has been cloned from Phalaris coerulescens. This gene is characterized by two major domains: an allele specificity domain and a thioredoxin catalytic domain. A family of sequences with varying degrees of homology to this gene has been identified among 15 grass species covering all subfamilies of the Poaceae. These S-related sequences appear to be present in the grass family regardless of self-compatibility. Evidence is presented to show that at least one of the sequences is transcribed, suggesting a functional gene. In contrast to the high expression of the S gene in Phalaris pollen, expression of the related gene in the pollen (or anthers) of the grass species examined was so low that RNA gel blot analysis failed to display a significant signal. However, reverse transcription-based polymerase chain reaction (RT-PCR) successfully amplified the region corresponding to the S thioredoxin domain from 10 of the grass species. With grasses other than Phalaris, RT-PCR showed limited success in amplifying the region corresponding to the S variable portion at the 5′ end of the Phalaris S gene. Sequencing of the PCR-amplified S thioredoxin region from wheat, barley, rye and Dactylis revealed that this is a highly conserved gene with 94–97% sequence similarity with the corresponding Phalaris S gene. The conservation of sequence and ubiquitous expression of the gene across the grass family strongly suggest that the S-related gene is carrying out a significant biological function in the Poaceae. On the basis of these findings, a model for the evolution of the S self-incompatibility gene in the grasses is proposed.
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  • 50
    ISSN: 1573-5028
    Keywords: chloroplast-derived trnH ; chloroplast genome ; DNA transfer ; gene expression ; Gramineae ; mitochondrial genome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We reported previously that the mitochondrial sequence that contains the chloroplast-derived trnH gene has been highly conserved in the region around one terminus of the junction between chloroplast-derived and mitochondrion-specific sequences in most of the gramineous plants analyzed [15]. The results of RT-PCR, northern hybridization, in vitro capping and ribonuclease protection experiments show that the chloroplast-derived trnH gene is transcribed from a putative promoter that is located in the mitochondrion-specific sequence. Gene expression in this region seems to be correlated with the conservation of the sequence at the junction between the chloroplast-derived fragment and the mitochondrion-specific sequence.
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  • 51
    ISSN: 1573-5028
    Keywords: Bacillus thuringiensis ; coleoptera ; eggplant ; gene expression ; insect resistance ; Solanum integrifolium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Solanum melongena (eggplant) cv. Picentia and the wild species Solanum integrifolium were transformed with both a wild type (wt) and four mutagenized versions of Bacillus thuringiensis (Bt) gene Bt43 belonging to the cry3 class. The Bt gene was partly modified in its nucleotide sequence by replacing four target regions (W: +1 to +170; X: +592 to+1057 ; Y: +1203 to +1376; Z: +1376 to +1984) with synthetic fragments obtained by polymerase chain reaction amplification of crude oligonucleotides. The synthetic Bt genes were designed to avoid, in their modified regions, sequences such as ATTTA sequence, polyadenylation sequences and splicing sites, which might destabilize the messenger RNA. Furthermore, the codon usage was improved for a better expression in the plant system. The amino acid composition was not altered. Four versions of the modified Bt gene were obtained, BtE, BtF, BtH and BtI, with a nucleotide subtitution percentage of 8.2, 8.6, 14, and 16%, respectively, in comparison to the wt gene Bt43. Modified versions contained different subsets of substituted regions: BtE - W+Z, BtF - Y+Z, BtH - X+Y+Z, BtI - W+X+Y+Z. In the final modified version (BtI), overall guanine + cytosine was increased from the 34.1% of the wt gene to 45.5%, and most of the destabilizing sequences were eliminated. Transgenic plants obtained with the more modified versions, BtH and BtI, were fully resistant to Leptinotarsa decemlineata Say first- and third- instar larvae, while Bt43 wt, BtE and BtF genotypes did not cause mortality and did not affect larval development.
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  • 52
    ISSN: 1573-5028
    Keywords: chlorophyll a-binding protein ; gene expression ; LHC ; light-harvesting complex ; photosystem I ; rhodophyte
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The accessory light-harvesting polypeptides associated with photosystem I (LHCI) in Porphyridium cruentum bind chlorophyll a, zeaxanthin and β-carotene. A cDNA library of P. cruentum was screened with an antiserum specific to the LHCI polypeptides, and an 0.9 kb fragment was identified as coding for an LHCI polypeptide. This cDNA, which we named LhcaR1, has an open reading frame encoding 222 amino acid residues including a putative transit peptide of 28 amino acids. Hydropathy analysis suggests that there are three transmembrane helices in the mature polypeptide. Each of the amino acid residues that bind chlorophyll (six residues) and serve in stabilizing the helices in higher-plant LHCs are conserved in helices 1 and 3 of P. cruentum LhcaR1. The N-terminal flanking regions of these two helices also show high sequence conservation with other LHCs. Helix 2 contains a seventh putative chlorophyll-binding site, but resembles helix 2 of higher-plant LHCs to a lesser degree. A sequence motif of 11 residues found near the N-terminus and in each of the three helices suggests the possibility that the red algal LhcaR1 derives from a gene duplication. Polypeptides of the expected molecular weight in six other red algae (Achrochaetium, Bangia, Callithamnion, Cyanidium, Polysiphonia, Spermothamnion) were recognized by the antiserum to P. cruentum LHCI, indicating a wide distribution of LHCI in rhodophytes.
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  • 53
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    Plant molecular biology 33 (1997), S. 1-10 
    ISSN: 1573-5028
    Keywords: androgenesis ; anther and microspore culture ; gene expression ; haploids ; pollen embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 54
    ISSN: 1573-5028
    Keywords: 1-aminocylopropane-1-carboxylate ; ACC oxidase ; ACC synthase ; cold ; 1-methylcyclopropene ; propylene ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Passe-Crassane pears require a 3-month chilling treatment at 0 °C to be able to produce ethylene and ripen autonomously after subsequent rewarming. The chilling treatment strongly stimulated ACC oxidase activity, and to a lesser extent ACC synthase activity. At the same time, the levels of mRNAs hybridizing to ACC synthase and ACC oxidase probes increased dramatically. Fruit stored at 18 °C immediately after harvest did not exhibit any of these changes, while fruit that had been previously chilled exhibited a burst of ethylene production associated with high activity of ACC oxidase and ACC synthase upon rewarming. ACC oxidase mRNA strongly accumulated in rewarmed fruits, while ACC synthase mRNA level decreased. The chilling-induced accumulation of ACC synthase and ACC oxidase transcripts was strongly reduced when ethylene action was blocked during chilling with 1-methylcyclopropene (1-MCP). Upon rewarming ACC synthase and ACC oxidase transcripts rapidly disappeared in 1-MCP-treated fruits. A five-week treatment of non-chilled fruits with the ethylene analog propylene led to increased expression of ACC oxidase and to ripening. However, ethylene synthesis, ACC synthase activity and ACC synthase mRNAs remained at very low level. Our data indicate that ACC synthase gene expression is regulated by ethylene only during, or after chilling treatment, while ACC oxidase gene expression can be induced separately by either chilling or ethylene.
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  • 55
    ISSN: 1573-5028
    Keywords: proline ; Δ1-pyrroline-5-carboxylate synthetase ; osmotic stress ; gene expression ; salt tolerance ; rice (Oryza sativa L.)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA for Δ1-pyrroline-5-carboxylate (P5C) synthetase (cOsP5CS), an enzyme involved in the biosynthesis of proline, was isolated and characterized from a cDNA library prepared from 14-day-old seedlings of Oryza sativa cv. Akibare. The deduced amino acid sequence of the P5CS protein (OsP5CS) from O. sativa exhibited 74.2% and 75.5% homology to that of the P5CS from Arabidopsis thaliana and Vigna aconitifolia, respectively. Northern blot analysis revealed that the gene for P5CS (OsP5CS) was induced by high salt, dehydration, treatment of ABA and cold treatment, while it was not induced by heat treatment. Simultaneously, accumulation of proline was observed as a result of high salt treatment in O. sativa. Moreover, the levels of expression of OsP5CS mRNA and content of proline under salt stress condition were compared between a salt-tolerant cultivar, Dee-gee-woo-gen (DGWG) and a salt-sensitive breeding line, IR28. It was observed that the expression of the P5CS gene and the accumulation of proline in DGWG steadily increased, whereas those in IR28 increased slightly.
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  • 56
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    Plant molecular biology 33 (1997), S. 467-481 
    ISSN: 1573-5028
    Keywords: Ca2+ ; cell signaling ; Chlamydomonas ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Acid shock of Chlamydomonas results in flagellar excision and induction of flagellar protein RNAs. The magnitude of flagellar RNA accumulations after flagellar excision by mechanical shear depends on the extracel]ular Ca2+ concentration. In this report, we demonstrate that the magnitude and duration of flagellar RNA accumulations are signaled by an acid shock-induced Ca2+ influx. RNA accumulations were greater in cells acid shocked in 500 µM CaCl2 than in 200 µM CaCl2, although the accumulation durations were similar. RNA accumulations of lower magnitude and shorter duration were observed in cells in Ca2+-containing buffer treated with CdCl2. RNA accumulations were of still lower magnitude and shorter duration in cells shocked in buffer without added CaCl2 than in cells shocked in 200 or 500 µM CaCl2 or in the presence of CdCl2. RNA accumulations similar to those in cells shocked in buffer without added CaCl2 were measured in cells following acid shock in buffer containing 200 µM CaCl2 and supplemented with neomycin, ruthenium red, or LaCl3. Acid shock of the adf-1 mutant resulted in RNA accumulations of shorter duration and lower magnitude than those measured in adf-1 cells stimulated by mechanical shear. These results are consistent with an hypothesis that acid shock generates two genetically and pharmacologically distinct signals governing flagellar RNA induction; the first signal is independent of a Ca2+ influx and flagellar excision and results in low magnitude accumulations of short duration, and the second is a consequence of a Ca2+ influx and results in accumulations of high magnitude and long duration.
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  • 57
    ISSN: 1573-5028
    Keywords: 1-aminocyclopropane-1-carboxylate (ACC) synthase ; elicitor ; ethylene ; gene expression ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The key enzyme of ethylene biosynthesis, ACC synthase, is encoded by a multigene family. We describe three new DNA sequences encoding members of the ACC synthase family of the tomato. One of these sequences encodes a novel ACC synthase, LE-ACS6, which is phylogenetically related to the ACC synthases LE-ACS1A and LE-ACS1B. Gene-specific probes for seven tomato ACC synthase genes were prepared. They were used for RNase protection assays to study the accumulation of ACC synthase transcripts in suspension-cultured tomato cells after the addition of an elicitor. The ACC synthase genes LE-ACS2, LE-ACS5 and LE-ACS6 were strongly induced by the elicitor. In contrast, the genes LE-ACS1B, LE-ACS3 and LE-ACS4 were constitutively expressed and LE-ACS1B was present at all times at a particularly high level. Thus, there are two groups of ACC synthase transcripts expressed in these cells, either elicitor-induced or constitutive. A transcript of LE-ACS1A was not detected. Despite the presence of LE-ACS1B, LE-ACS2, LE-ACS3, LE-ACS4 and LE-ACS5, there was only little ethylene produced in the absence of the elicitor. Increased ethylene production is usually correlated with the accumulation of ACC synthase transcripts, indicating that ethylene production is controlled via the transcriptional activation of ACC synthase genes. However, the abundance of several ACC synthase mRNAs studied was not strictly correlated with the rate of elicitor-induced ethylene production. Our data provide evidence that the activity of these ACC synthases may not solely be controlled by the transcriptional activation of ACC synthase genes.
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  • 58
    ISSN: 1573-5028
    Keywords: conglutin ; gene expression ; leginsulin ; Lupinus angustifolius ; basic 7S globulin ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of genes encoding conglutin γ and a leginsulin-like protein has been examined in narrow-leafed lupin, Lupinus angustifolius L. Conglutin γ is a homologue of basic 7S globulin (Bg), the insulin and leginsulin binding protein from soybean. Accumulation of conglutin γ mRNA, as assessed by northern assays and reverse-transcription PCR, was tightly regulated both spatially and temporally in lupin plants and was detected almost exclusively in developing seeds. Similar tissue and temporal specificity was demonstrated when 1.8 kb of the promoter region from the conglutin γ gene was used to drive the expression of a β-glucuronidase reporter gene in transgenic plants. In stably transformed tobacco the conglutin γ promoter produced strong, temporally regulated and seed-specific expression of the reporter gene which was localised to the embryo tissues and to a layer of cells adjacent to the seed coat. A truncated 0.29 kb promoter fragment produced much reduced levels of expression and a loss of embryo specificity. Leginsulin-like mRNA was similarly detected in lupins only in developing seeds. The leginsulin-like gene detected in L. angustifolius showed 96% sequence identity to leginsulin from soybean within the 280 bp region amplified from lupin by PCR. The results demonstrate that both components of a Bg-leginsulin putative signal transduction pathway are present in the seeds of lupin.
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  • 59
    ISSN: 1573-5028
    Keywords: auxin ; cell expansion ; cellulase ; endo-1,4-β-glucanase ; ethylene ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plant developmental processes involving modifications to cell wall structure, such as cell expansion, organ abscission and fruit ripening, are accompanied by increased enzyme activity and mRNA abundance of endo-1,4-β-glucanases (EGases). An EGase cDNA clone, Ce14, isolated from tomato (Lycopersicon esculentum) has been shown to be identical to a tomato pistil-predominant EGase cDNA, TPP18. In addition to its previously reported expression during certain stages of early pistil development, Ce14 mRNA was also detected at high levels in the growing zones of etiolated hypocotyls (about 2.5-fold less than in pistils) and in young expanding leaves (about 3.5-fold less than in pistils). The abundance of Ce14 mRNA declined precipitously in older tissues as cells became fully expanded, and was barely detectable in mature vegetative tissues. Ce14 mRNA abundance was also low in abscission zones, and did not increase as abscission progressed. In fruit, Ce14 mRNA was present at low levels during fruit expansion, but was essentially absent during subsequent fruit development and ripening. Treatment of etiolated hypocotyls with ethylene or high concentrations of auxin sufficient to induce rapid lateral cell expansion and hypocotyl swelling also brought about an approximate doubling of Ce14 mRNA abundance, suggesting that Ce14 mRNA accumulation may be promoted directly or indirectly by ethylene. Thus, accumulation of Ce14 mRNA was found to be correlated with rapid cell expansion in pistils, hypocotyls and leaves.
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  • 60
    ISSN: 1573-5028
    Keywords: gene expression ; gene family ; higher plant ; phosphorylation ; post-transcriptional regulation ; Solanum tuberosum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A polymerase chain reaction product (PKIN503) was amplified from potato (Solanum tuberosum) cv. Désirée using oligonucleotide primers with sequences which are highly conserved in the plant sucrose non-fermenting 1 (SNF1)-related protein kinase gene family. Southern blot analysis showed the presence of 5–10 SNF1-related genes in the potato genome. PKIN503 was used to screen a tuber cDNA library and a genomic library, and one cDNA and five genomic clones were isolated. The nucleotide sequences of a portion of all five genomic clones were shown to be identical and only one, pgPKIN1, was analysed further. The cDNA was found to be truncated at the 5′ end but the cDNA and genomic sequences contained only 15 substitutions, two of which resulted in changes in the derived amino acid sequence. PKIN1 was shown to encode an Mr 57854 protein with 61–70% sequence similarity with other plant SNF1-related protein kinases. Northern blot analysis revealed some tissue-specific differences in PKIN1 transcript levels, the lowest being detected in leaves and the highest in stolons. However, much greater differences were found in SNF1-related activity, which was measured using a phosphorylation assay with a substrate peptide which has been shown previously to be phosphorylated by plant SNF1-related protein kinases. Activity decreased by almost 80% during development from stolons to mature tubers but it increased about seven-fold during the first seven days of storage after harvesting, before decreasing again. However, activity was highest in mini-tubers, where the levels were 37 times greater than those in mature tubers from a pot-grown plant. Transcript levels in these tissues were approximately equal, clear evidence that SNF1-related protein kinase activity in potato is regulated, in part, post-transcriptionally.
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  • 61
    ISSN: 1573-5028
    Keywords: gene expression ; gibberellin biosynthesis ; gibberellin 20-oxidases ; Phaseolus vulgaris ; Pisum sativum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract PCR was used with degenerate primers based on conserved amino acid sequences in gibberellin (GA) 20-oxidases to isolate cDNA clones for these enzymes from young seeds of pea (Pisum sativum) and developing embryos of French bean (Phaseolus vulgaris). One GA 20-oxidase cDNA (Ps27-12) was obtained from pea and three (Pv15-11, Pv73-1 and Pv85-26) from bean. Their identities were confirmed by demonstrating that fusion proteins expressed in Escherichia coli exhibited GA 20-oxidase activity, converting [14C]GA12 to [14C]GA9. The intermediates in this three-step reaction, GA15 and GA12, were also identified as products. The expression proteins from three of the clones (Ps27-12, Pv15-11 and Pv73-1) were also shown to convert GA53 to GA20, as effectively as they did GA12. On the basis of transcript levels measured by northern blot analysis, the pea GA 20-oxidase gene is most highly expressed in young leaves, fully expanded internodes, very young seeds (until 4 days after anthesis) and expanding pods (from 3 days after anthesis at least until day 6). Expression in pods from 3-day-old unpollinated ovaries is higher than in those from pollinated ovaries. Treatment of unpollinated ovaries with GA3 to induce parthenocarpic fruit-set severely reduced the amount of GA 20-oxidase mRNA, whereas treatment with 2,4-D, although inducing fruit-set, did not reduce the levels of these transcripts. Plant decapitation above an unpollinated ovary resulted in very high levels of GA 20-oxidase mRNA in the pod. The three GA 20-oxidase genes from French bean showed very different patterns of expression: Pv15-11 was expressed in the roots, young leaves, and developing seeds, but most highly in immature cotyledons, while Pv73-1 has a similar expression pattern to Ps27-12, with transcripts found only in young seeds and young leaves, where it was particularly abundant. Transcripts corresponding to Pv85-26 were detected in developing seeds, and just traces in the young leaves. Southern blot analysis indicated that the bean GA 20-oxidases are each encoded by single-copy genes, whereas one more gene, homologous to Ps27-12, could also exist in pea.
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  • 62
    ISSN: 1573-5028
    Keywords: cDNA cloning ; environmental factors ; gene expression ; leaf senescence ; organs ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Senescence-related cDNA clones designated SENU1, 4, 5 (senescence up-regulated) and SEND32, 33, 34, 35 and 36 (senescence down-regulated) isolated from a tomato leaf cDNA library [9] were characterized. Southern analysis showed that SEND32 is encoded by a single-copy gene while SEND33, 34, 35, 36 and SENU1 and SENU5 are members of small gene families. DNA and protein database searches revealed that SEND32, SEND35, SENU1 and SENU5 are novel cDNAs of unknown function. SEND33 encodes ferredoxin, SEND34 encodes a photosystem II 10 kDa polypeptide and SEND36 encodes catalase. The SENU4 sequence is identical to the P6 tomato protein previously reported to be pathogenesis-related [46]. The mRNA levels of SENU1, 4 and 5 increased during leaf senescence and SENU1 and SENU5 were also expressed at high levels during leaf development and in other plant organs. The SENU4 mRNA was associated more specifically with leaf senescence, although low expression was also detected in green fruit. The mRNAs for all SEND clones decreased during tomato leaf development and senescence and all except SEND32 were expressed at low levels in other plant organs. The accumulation of mRNA homologous to SENU4 and the decrease in abundance of SEND32 provide good molecular markers for leaf senescence.
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  • 63
    ISSN: 1573-5028
    Keywords: S-adenosylmethionine decarboxylase ; carnation ; cDNA sequence ; gene expression ; protein processing ; uORF
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract S-adenosylmethionine decarboxylase (SAMDC; EC 4.1.4.50) is one of the key enzymes in polyamine biosynthesis, and the product of its catalytic reaction, decarboxylated S-adenosylmethionine (dcSAM), serves as an aminopropyl donor in the biosynthesis of spermidine and spermine. In order to provide information on the structure and regulation of SAMDC, we have isolated and sequenced two different SAMDC cDNA clones from carnation petals. The nucleotide sequences of CSDC9 and CSDC16 show 78.3% identity, and the deduced amino acid sequences show 81.7% identity and 86.5% similarity [12]. There are several regions with highly conserved sequences among SAMDC cDNAs of potato, spinach, periwinkle, man and yeast. These conserved regions include a cleavage site for the processing of SAMDC proenzyme and a putative PEST sequence that may be relevant to the rapid degradation of SAMDC protein. Carnation SAMDC cDNAs have long transcript leaders of 472 bp and 502 bp for CSDC9 and CSDC16, respectively. Both sequences contain short upstream open reading frames (uORFs) in their 5′ -untranslated regions. The CSDC9 uORF is 54 amino acids from 152 to 317 while the corresponding sequence in CSDC16 is 52 amino acids located from 156 to 314 in each 5′-untranslated region. The nucleotide sequences of uORFs in CSDC9 and CSDC16 were 89.9% identical. In vitro transcription/translation experiments showed: (1) each proenzyme of both cDNAs of SAMDC was converted to two polypeptides consisting of a large subunit (calculated as 31544 Da and 32537 Da, respectively) and a small subunit (calculated as 9704 and 9041 Da, respectively) after 20 min of translation; (2) the processing occurs rapidly during the translation of protein. But once the translation process is stopped accumulation of the subunits slows and never reaches completion even after 300 min. The processing of carnation SAMDC enzyme is not stimulated by putrescine in in vitro transcription/translation reaction.
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  • 64
    ISSN: 1573-5028
    Keywords: arginine decarboxylase ; gene expression ; Oryza sativa ; polyamines ; rice ; salinity stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of salinity stress on the activity of arginine decarboxylase (ADC, EC 4.1.1.19), the first enzyme in biosynthesis of polyamines (PA) from arginine, as well as its transcript level has been compared in salt-sensitive (M-1-48) and salt-tolerant (Pokkali) rice cultivars. Treatment of 72 h grown seedlings either with increasing concentrations of NaCl or with 150 mM NaCl for different time periods, showed a gradual increase of activity in Pokkali. In M-1-48 an immediate increase followed by sharp decrease was observed on prolonged treatment beyond 6 h or above 150 mM NaCl. To generate a DNA probe for ADC, the polymerase chain reaction was used with oat genomic DNA and sequence-specific primers. A region of oat genomic DNA containing a coding sequence for 166 amino acids of the C-terminal part of the ADC enzyme was amplified and called OAD1. Southern analysis of EcoRI- or BamHI-cut genomic DNAs from different cultivars of rice with OAD1 as the probe revealed strong hybridization with one DNA fragment of rice and restriction fragment length polymorphism (RFLP) was noticed. Northern analysis of total RNA of rice with OAD1 as the probe revealed hybridization with a transcript of similar size to the ADC transcript in oat. While in Pokkali, at least a 20-fold accumulation of OAD1 homologous transcript was detected after treatment with 200 mM NaCl, only a seven-fold increase in transcript level was found in M-1-48 after 150 mM NaCl treatment. Results suggest that in the salt-tolerant rice cultivar Pokkali, ADC enzyme activity increases and its transcript also accumulates during the prolonged salinity stress, this mechanism is absent in the salt-sensitive rice cultivar M-1-48 where a prolonged period of salinity stress down-regulates both ADC activity and its transcript level.
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  • 65
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    Plant molecular biology 34 (1997), S. 643-650 
    ISSN: 1573-5028
    Keywords: callus ; crown tissue ; gene expression ; low temperature ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The low-temperature (2 °C)-specific wheat cDNA, pTACR7, represents a gene designated tacr7 from hard red winter wheat (HRWW; Triticum aestivum L. cv. Winoka). The term low-temperature-specific (LTS) is used because tacr7 is not induced by ABA or stresses such as salt, dehydration, and heat. pTACR7 was isolated by RT-PCR with mRNA from wheat crown tissue, the oligonucleotide primers derived from the barley cognate pHVCR8 (GenBank accession number L28091). Based on the deduced amino acid sequence, TACR7 is highly hydrophobic, with a single transmembrane domain and an amino acid bias for leucine (19%). Thus, the encoded protein TACR7 is unique among low-temperature-regulated wheat proteins described in the literature. Analysis of steady-state levels of tacr7 transcripts (630 nt) showed accumulation in wheat seedlings, crown tissue, and callus cultures after transfer from control (25 °C) to low temperature (2 °C). No detectable transcripts were observed by northern blot hybridization with pTACR7 probe from seedling or callus treated with ABA, salt, dehydration, or heat stress. tacr7 transcripts accumulated during 2 °C exposure to a greater amount in a freeze-resistant HRWW (FR; SDmut 16029) than in a freeze-susceptible HRWW (FS; SDmut 16169) crown tissue, with the largest difference between genotypes being 30% ± 3% at 3 weeks.
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  • 66
    ISSN: 1573-9368
    Keywords: peanut ; engineered virusresistance ; tomato spotted wiltvirus ; transformation ; nucleocapsid ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucleocapsid gene of tomato spotted wilt virus Hawaiian L isolate in a sense orientation, and the GUS and NPTII marker genes, were introduced into peanut (Arachis hypogaea cv. New Mexico Valencia A) using Agrobacterium-mediated transformation. Modifications to a previously defined transformation protocol reduced the time required for production of transformed peanut plants. Transgenes were stably integrated into the peanut genome and transmitted to progeny. RNA expression and production of nucleocapsid protein in transgenic peanut were observed. Progeny of transgenic peanut plants expressing the nucleocapsid gene showed a 10- to 15-day delay in symptom development after mechanical inoculations with the donor isolate of tomato spotted wilt virus. All transgenic plants were protected from systemic tomato spotted wilt virus infection. Inoculated non-transformed control plants and plants transformed with a gene cassette not containing the nucleocapsid gene became systemically infected and displayed typical tomato spotted wilt virus symptoms. These results demonstrate that protection against tomato spotted wilt virus can be achieved in transgenic peanut plants by expression of the sense RNA of the tomato spotted wilt virus nucleocapsid gene
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  • 67
    ISSN: 1573-9368
    Keywords: superoxide dismutase ; recombinant protein ; transgeneexpression ; metalloprotein ; gene expression ; transgenic rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Expression of human extracellular superoxide dismutase (EC-SOD), a glycosylated, tetrameric metalloprotein, was targeted to the lactating mammary gland of transgenic rabbits. Efficient expression of the recombinant whey acidic protein/ec-sod gene was achieved and up to 3 mg ml−1 of the enzyme was secreted into the milk. Rabbit milk-produced recombinant EC-SOD was primarily found in the whey and purified by a two-step chromatographic method. To evaluate the rabbit milk-produced human EC-SOD, comparisons with native and Chinese hamster ovary cell (CHO)-produced EC-SOD were performed. All proteins were tetrameric and N-glycosylated. The behaviour on SDS-PAGE and size-exclusion chromatography indicated that the masses, and thereby the extent of post-translational modification of the proteins was similar. The monosaccharide composition of both recombinant EC-SOD variants was analysed and indicated similarities in the attached N-glycans on the two proteins. Furthermore, the peptide maps of the three EC-SOD variants revealed that all proteins had similar polypeptide backbones
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  • 68
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    Journal of applied phycology 9 (1997), S. 19-24 
    ISSN: 1573-5176
    Keywords: Chaetoceros ; growth ; neutral lipid ; total lipid ; nitrogen deprivation ; Nile Red
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chaetoceros muelleri (Schütt) was cultured on a thermal gradient plate, subjected to two media types with a range of specific conductances, and evaluated for growth and neutral lipid accumulation. Growth was measured directly by daily changes in cell numbers and indirectly by changes in optical density at 750 nm. C. muelleri exhibited a growth rate of at least two doublings day-1 over broad temperature (20 to 35 °C) and conductance ranges (10 to over 60 mS cm-1) and the optimum growth rate approached 4.0 doublings day-1 at 30 °C and a conductance of 25 mS cm-1. Intracellular neutral lipid storage was evaluated with fluorometry and epifluorescent microscopy using the fluorochrome Nile Red. Gravimetric analysis revealed a total lipid content in nitrogen-depleted cultures of C. muelleri of over 400 mg L-1, five to seven times that observed in nitrogen-replete cultures. Based on its high growth rate, tolerance to a broad range of temperatures and specific conductances, and large quantity of intracellular lipid, C. muelleri may have potential for exploitation as a renewable precursor to liquid fuels or as a lipid source.
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  • 69
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    Journal of applied phycology 9 (1997), S. 437-444 
    ISSN: 1573-5176
    Keywords: Haematococcus pluvialis ; experimental design ; optimization ; culture medium ; growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A central composite rotatable design was used to examine the effects of five components of the medium on the growth of Haematococcus pluvialis in batch culture. The medium components considered were: sodium acetate,potassium nitrate, major elements, trace elements and vitamins. Within the range of the concentrations tested, a moderate concentration of the major elements significantly enhanced algal growth, both in terms of specific growth rate and cell dry weight, whereas the vitamins had no significant effect. Based on the response surface contour plots and the results of numerical analyses, the optimal nutrient concentrations for growth in terms of specific growth rate were 0.51 g L-1 sodium acetate, 0.25 g L-1 potassium nitrate, 0.63 mL L-1 of the major element stock solution and 0.2 mL L-1 of the trace element stock solution. The optimal nutrient concentrations for biomass production were 1.64 g L-1 sodium acetate, 0.37 g L-1potassium nitrate, 2.52 mL L-1 of the major element stock solution and 0.03 mL L-1 of the trace element stock solution.
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  • 70
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    Journal of applied phycology 9 (1997), S. 551-558 
    ISSN: 1573-5176
    Keywords: Spirulina ; Arthrospira ; growth ; salinity ; quantum yield ; nutrient status ; Chitu ; Ethiopia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Spirulina platensis (=Arthrospira fusiformis) was isolated from Lake Chitu, a soda crater lake in the Ethiopian Rift Valley, where it formsa dense and almost unialgal population. Growth experiments were run in turbidostats under constant light, to assess growth response and tolerance to salinity, as well as to the component anions. Salinity was tested over the range 13–88 g L-1 using additions of NaHCO3, NaCl or Na2SO4. A maximum specific growth rate (µmax d-1) of 2.14 was achieved at the lowest salinity, but quantum yield (Φ%) was highest between 33 to 51 g L-1. Increasing salinity of the medium reduced the specific growth rate (µ) to a minimum of 0.33 d-1, and Φ to 〈 0.5%. Growth response in terms of µ and Φ was best in HCO 3 - , less in Cl-, and least in SO 4 2- series. Cultures showed obvious differences in cellular morphology, pigment, nitrogen and phosphorus contents in response to treatment with the different anions. Results indicate that the species has a wide range of tolerance to salinity from NaHCO3. Some degree of tolerance is also shown to high concentrations of Cl- and SO 4 2 , but with an overall lower performance of cells in terms of growth rate, light utilization efficiency, and nutrient status to cells grown in high HCO 3 - concentrations and the same levels of salinity and light.
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  • 71
    ISSN: 1572-9710
    Keywords: ancient woodland indicator species ; conservation ; management ; restoration ; soil seed bank
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A survey was carried out to determine the density and species composition of germinable seed in the surface soil layers of 30 plots within a coniferized lowland woodland in East Kent in order to establish the resources available for habitat creation in the event of some areas being modified during a proposed holiday village development. The selected plots included conifer plantations (up to 69 years old), broad-leaved plantations and the semi-natural broad-leaved edges that remain on parts of the perimeter of the site which were used as the control. A total of 13 682 seedlings emerged from the soil samples during the four-month germination trials. Fifty-two species were identified of which eight were ancient woodland indicator species for south-east England. The most abundant species represented in the seed banks were: Juncus effusus, Rubus fruticosus, Carex sylvatica, Betula pendula and Agrostis tenuis. Between-site comparisons of coniferous plots of different ages revealed a marked reduction in the seed species and seed density in plantations over 65 years old. Results of soil nutrient and texture analyses ruled out the likelihood of edaphic factors being responsible for between-site differences in seed bank composition. Seed species richness and diversity (Shannon-Wiener diversity index) were greatest in the semi-natural broad-leaved edges, but the diversity index used also showed that two replanted conifer sites had high values despite few species being present. The usefulness and limitations of diversity indices in the context of seed bank studies is discussed. From the results of the study, management proposals for the site have been put forward in order to maintain floristic diversity and mitigate the impact of the proposed development.
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  • 72
    ISSN: 1572-9710
    Keywords: biodiversity ; conservation ; development ; planning ; ecodevelopment ; participatory ; India
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The existing network of Protected Areas (PAs) in India is the major effort aimed at biodiversity conservation at the national level. The sustainability of PAs is heavily influenced by local people who are largely dependent on natural resources (fuel, fodder, minor forest products) for their livelihood. While all PAs are surrounded by historically resource-dependent communities, several of them have villages within their core areas, too. This has necessitated an alternative approach to natural resource management which aims to integrate the interests of conservation with those of the nearby resource dependent communities. The case of the Great Himalayan National Park illustrates and incorporates the lessons from Integrated Conservation and Development Projects (ICDPs) implemented elsewhere in the world.
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  • 73
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    Biodiversity and conservation 7 (1997), S. 109-135 
    ISSN: 1572-9710
    Keywords: New Caledonia ; endemism ; plants ; conservation ; protected areas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract With 76% of its 3063 native species of flora endemic, the New Caledonia biodiversity hotspot has long been recognized as having a high potential for conservation. Under the new IUCN Red List categories, 25% of the endemic plants are at risk (Conservation Dependent, Vulnerable, Endangered, Critically Endangered), and five species are already extinct. A review of their distribution demonstrates that 83% of the threatened species do not occur at all in a conservation area, and only 11% have their conservation status improved by a protected area. The protected area network is geographically and floristically very unbalanced, with the rainforest and high altitude maquis in the south concentrating most of the conservation effort. Conversely, the middle and northern segments of the island, as well as all of the dry west coast, are left without adequate conservation area. Two vegetation types, the sclerophyll forest and the unique low/middle altitude maquis, are virtually totally unprotected. We conclude that the current network of protected areas needs to be considerably expanded, in terms of both geographical/floristic subregions within New Caledonia and vegetation type covered. With only 54% of the conservation area covered by strict mining restrictions, existing reserves need to have their conservation efficiency improved by a more vigorous enforcement of their status, and by extending mining bans to all of them.
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  • 74
    ISSN: 1573-5125
    Keywords: Marenzelleria ; Polychaetes ; macrozoobenthos ; abundance ; biomass ; population dynamics ; growth ; secondary production ; Baltic Sea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The propagation of an immigrant from North America, viz. the spionid Marenzelleria cf. viridis in the Darss-Zingst Bodden (DZB) (southern Baltic Sea), was studied at three stations from March 1992 to December 1995. Highest mean abundances (over 28 000 ind. m-2) and wet weights (400 g m-2) were recorded at station 2 in 1994. The spionid also reached its highest dominances in terms of biomass (40 to 90%) at this station, which was selected for the population dynamics, growth and secondary production studies. The spionid has a life span of about 3 years, and many individuals achieved sexual maturity after one year. Their growth curve is steepest during the first year of life, during which the animals grow to a length of about 180 segments. However, growth depression was observed during the ripening of the gametes in April, May and June. Secondary production was in the region of 55 to 85 g AFDW m-2 y-1. Productivity (P/B) varied considerably from generation to generation, ranging between 0 and 4.8 with an average between 1.2 and 1.6.
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  • 75
    ISSN: 1572-9710
    Keywords: Kenya ; birds ; conservation ; protected areas.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Patterns and environmental correlates of species distributions and richness are identified for Kenyan birds at a quarter degree-square scale. This information is used together with iterative complementarity analyses, which employ species richness, taxonomic dispersion and range-restrictedness, to identify priority areas for possible conservation attention. Bird species apparently not conserved by existing protected areas in Kenya are identified. Six avifaunal zones (and one transitional zone) are distinguished based on distributions of suites of bird species. Variation in biotope diversity (the number of forest and aquatic systems) accounts for 79% of the observed variation in Kenyan bird species richness. Although both rainfall and altitudinal range are significantly correlated with species richness, they only explain an additional 3% of the observed variation. The priority areas identified are situated mainly within highlands and coastal lowlands. Although few priority areas are identified in northern Kenya, this region also constitutes a priority, as it contains a suite of xeric species with habitats that are not represented elsewhere in Kenya. The papyrus yellow warbler, Chloropeta gracilirostris, William's bush lark, Mirafra williamsi, white-winged dove, Streptopelia reichenowi, and Jubaland weaver, Ploceus dichrocephalus, are identified as endemics or near-endemics that are probably not adequately conserved in Kenya at present.
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    Biodiversity and conservation 6 (1997), S. 853-868 
    ISSN: 1572-9710
    Keywords: protected area network ; biodiversity ; conservation ; North-East India ; Meghalaya
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The North-Eastern region of India is significant for biodiversity conservation because of its floristic richness and high levels of endemism. Deforestation levels are high in the region due to anthropogenic pressures. We accessed various literature sources to create a database for Meghalaya state containing information on plant species, habit, altitudinal distribution, endemism, and endangered status. Information on the existing protected area network (type, extent, and altitudinal representation) was added to the database. The database was used to assess the effectiveness of the existing protected area network in conserving the floristic biodiversity of the state. Of a total of 3331 plant species, 1236 (37.11%) are endemic of Meghalaya and 133 (4%) are confined to 'sacred forests'. However, 'sacred forests' are not legally protected areas. Only 32 220 ha (1.43% of the state's geographical area) is protected under the category of National Park or Sanctuary. Although 212 species (17.15% of the state's endemic species) occur only in Meghalaya at altitudes above 1500 m, none of the forests at these altitudes are protected as National Parks or Sanctuaries. We conclude that the existing protected area network does not effectively conserve the state's unique biodiversity and suggest measures by which its effectiveness might be increased.
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    Biodiversity and conservation 6 (1997), S. 1007-1026 
    ISSN: 1572-9710
    Keywords: West Africa ; Ghana ; conservation ; small mammals ; sacred groves ; interdisciplinary development ; biodiversity ; sustainability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This article considers the conservation of relict natural habitat in West Africa, especially habitat preserved in traditional sacred groves. Government-sanctioned conservation is contrasted with local grassroots efforts of conservation. Evidence for the ecological value of sacred groves is based on results of a field study of small mammal communities conducted on the Accra Plains of Ghana and on published sources on the conservation and use of sacred groves from various countries. The study employed standard mark-and-recapture techniques for the sampling for terrestrial small mammals, and mist netting for the sampling of bat communities. Pragmatic approaches combining conservation and sustainable use are considered, as are traditional values that have preserved the sacred groves in the past for up to several hundred years in some cases. In part because these groves shelter unique small mammal and plant communities, traditional values and protection mechanisms should be integrated into the newly emerging cultural and religious contexts. The issues encountered during this study reveal that effective conservation involving local peoples requires a concerted interdisciplinary effort.
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    Biodiversity and conservation 7 (1997), S. 97-107 
    ISSN: 1572-9710
    Keywords: Island biology ; introduced species ; Galá ; pagos ; conservation ; endangered plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Although still in relatively good condition, the Gala´pagos Archipelago suffers from increasing human pressures. Apart from direct actions like hunting and logging, endemic plants and animals are threatened by introduced species, and in many cases the present status of the populations is not known. The conservation status of eight plant species considered endangered was studied from literature and field surveys and the main threats were determined. Each of the eight species is endemic to only one island but in some cases is also present on nearby islets. Of these eight species, one is considered extinct, one critically endangered, and the others suffer various levels of threat. As in all island systems of the world, the main threats are introduced organisms, both plants and animals. The extinct species probably disappeared owing to invasion by Lantana camara, one of the most aggressively invasive plants of the islands, and the most endangered species is threatened by goats. The remaining species seem to be regenerating well and we can expect positive results from protection efforts. Today, only one of the eight species benefits from a direct protection action.
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  • 79
    ISSN: 1573-5125
    Keywords: Marenzelleria cf. viridis ; Polychaetes ; food limitation ; food selection ; growth ; gut passage time ; food quality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phytoplankton 〈 20 µm was a principal dietary component of the larvae of Marenzelleria cf. viridis. Maximum ingested particle size increased as animal size increased, reaching a maximum diameter of 80 µm for larvae with 6 to 10 setigers. The larvae started ingesting particulate matter at the 1-setiger stage and were able selectively to ingest phytoplankton and polystyrene particles of various sizes. Larvae in the 6 to 10-setiger size group did not differ from those in the 11 to 17-setiger size group in respect of size selectivity for polystyrene particles. The gut passage time for Chlorella vulgaris was ≥ 20 min. The ingestion rate was limited by food concentrations even at concentrations much higher than those encountered in the natural biotope, saturation being reached at a concentration of 28.5 times 106 cells ml-1 (117.7 mg C l-1. The low maximum filtration rate of only 1.19 µl ind.-1 h-1 indicates that the filtering capacity of the larvae is low. The larvae are still capable of food uptake at 1 °C. Further experiments demonstrated that larval growth and survival were strongly dependent on both food concentration and quality. Larval growth was food-limited under biotope conditions of the Darss–Zingst Boddens and even more so under Baltic Sea conditions. The results indicate that Marenzelleria cf. viridis is a species adapted to eutrophic conditions prevailing in brackish waters.
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  • 80
    ISSN: 1573-5176
    Keywords: Lyngbya wollei ; paralytic shellfish poisons ; growth ; toxin production
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Collections of Lyngbya wollei were taken from Guntersville Reservoir, Alabama, over a period of three years. Healthy filaments were isolated and transferred to agar plates of Z-8 and LM6E media. Unialgal isolates were cultured for the study of growth and paralytic shellfish poison (PSP) production. Filaments were extracted and the toxins were detected using high performance liquid chromatography (HPLC) with post column oxidation followed by fluorescence detection. HPLC profiles show that laboratory cultures of L. wollei produced decarbamoyl gonyautoxin 2 and 3, plus several other PSP like toxins whose structures are under investigation. At 26 °C and a light intensity of 11 or 22 µmol m-2 s-1 optimum production of both biomass and toxins occurred. A decrease or increase in temperature or light flux caused a reduction in dry weight or toxicity. Compared to control levels, lower PO4-P and NO3-N and higher calcium levels gave rise to higher biomass and toxicity. Lower calcium, calcium- or PO4-P deficient medium and high NO3-N or PO4-P caused a large decrease in dry weight and toxicity.
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    Aquarium sciences and conservation 1 (1997), S. 79-90 
    ISSN: 1573-1448
    Keywords: Freshwater fishes ; SoutheastAsia ; aquarium fish ; conservation ; biodiversity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract SoutheastAsia is an area rich in biodiversity, with a high degree ofendemism in both flora and fauna. Many freshwater fish specieshave been exploited for the ornamental fish trade. As thepopulation in South East Asia increases, vast tracts of forestsare cleared for agricultural, industrial and urbanizationpurposes. To conserve and sustainably exploit the wild fishpopulation, measures should be adopted to protect this naturalresource. Already, several ornamental species have been severelyoverexploited, e.g. bala shark (Balantiocheilos melanopterus),pygmy loach (Botia sidthimunki) and arowana (Scleropagesformosus), but the extirpation of local populations occurs formany reasons, including deforestation, and not just because offishing for the trade. There are also still many species thathave great ornamental fish potential. Wild fish species have alsobeen successfully bred in captivity and conserved, e.g. tigerbarb (Puntius tetrazona), bala shark, pygmy loach and arowana.Other methods of conservation include public education, leavingpristine forests intact and reforestation. The aquarium trade ofthe bala shark, harlequin rasbora (Rasbora heteromorpha), clownloach (Botia macracanthus), arowana and sawfish (Pristismicrodon) is discussed
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  • 82
    ISSN: 1573-1448
    Keywords: anaesthetic ; growth ; Haemulidae ; 2-phenoxyethanol ; Pomadasys commersonnii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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    Urban ecosystems 1 (1997), S. 103-116 
    ISSN: 1573-1642
    Keywords: wildlife ; greenways ; conservation ; corridors ; urban landscapes ; deer ; fox
    Source: Springer Online Journal Archives 1860-2000
    Topics: Architecture, Civil Engineering, Surveying , Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Goals related to wildlife conservation are stated or implied in many urban greenway plans, but the actual wildlife conservation value of urban greenways is unclear. We surveyed 72 km. of greenway consisting of 38 different greenway segments in six cities in the mid-southeastern United States to determine the presence or absence of red fox (Vulpes vulpes), grey fox (Urocyon cinereoargenteus), and white-tailed deer (Odocoileus virginianus), and to assess characteristics of the greenways and adjacent lands that might promote or limit the presence of these species in each greenway. We focused on fox and deer because these mammals, while habitat generalists, require diverse habitats within close proximity, and are more sensitive to human disturbance and have larger territories than the majority of urban wildlife in this region. Thus, they can serve as indicator species for these habitat values in urban greenways in the southeastern United States. Field observations and scent station inventories revealed that only 18 of the 38 greenway segments had either fox or deer, 12 had only fox, 6 had fox and deer, and none had deer alone. Greenway segments with more forest cover, wider corridors, greater amounts of adjacent natural or seminatural habitat, and forest connectivity between greenways and nonadjacent natural areas were more likely to have fox or deer present. Characteristics associated with fox and deer presence were not independent of each other. Because greenways with such characteristics tend to be surrounded by less urban areas, adjacent land use is a good predictor of fox and deer presence in the surveyed greenways. Whether greenways that are beneficial to fox and deer can be developed in urban settings is strongly site specific, because the greenway variables that appear most essential to these species are often dictated by the preexisting urban form and the vegetation in the vicinity of the greenway. Because the wildlife conservation value of urban greenways depends greatly on factors external to the greenway, greenways that have fox and deer present today may be particularly vulnerable to future changes in surrounding land use.
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  • 84
    ISSN: 1573-5117
    Keywords: microcrustacean zooplankton ; tropical limnology ; laboratory cultures ; growth ; development rates ; temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Length growth, instar durations, fecundity and mortality rates of fivespecies of microcrustacean zooplankton from a tropical reservoir weremeasured in relation to food quantity and temperature in laboratorycultures. Three cladocerans (Ceriodaphnia cornuta, Moina micrura,Diaphanosoma excisum), one calanoid copepod (Heliodiaptomus viduus), and onecyclopoid copepod (Mesocyclops thermocyclopoides) were studied. Filteredseston (45 µm mesh) from a local pond was used for food. Two foodconcentrations were employed: (1) 10 µg chlorophyll l−1(ca 0.25 mg Cl−1), and (2) 50 µg chlorophylll−1 (ca 1.25 mg C l−1). Food levels and watertemperature (22.5, 27.5, and 32.5 °C) used, roughly covered the rangesfound in the reservoir. Although all the three growth parameters were oftenaffected to some degree by temperature and food, the quantitative responseof the species differed. Also, the species reacted differently to the threepossible interactions (i.e. food × temperature,food × instar, andtemperature × instar). This contributed to the overalldifferences in the species responses. For the cladocerans, instar durationswere always affected by temperature. The food did not affect the durationtime of the adults and that of the combined juvenile instars, the latterexcept in one case significantly. For the two copepods food level affectedthe duration times of naupliar and copepodite instars, but the effect oftemperature was only significant for H. viduus. The development timesobserved for H. viduus were extraordinary long compared with values reportedin the literature for other tropical calanoids. This suggests that foodconditions in our culture were adversely affecting its growth rates.
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    Hydrobiologia 352 (1997), S. 287-293 
    ISSN: 1573-5117
    Keywords: Undaria pinnatifida ; translocation ; photoassimilates ; growth ; cultivation ; yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Undaria cultivation on a commercial scale began in China only in the last decade. Today, Undaria pinnatifida is the main species under cultivation concentrated in two provinces, Liaoning and Shandong. The annual production in the early nineties was 8000-13 000 tons dry weight, which is two or three times the pre-1980 figures. The raft cultivation method maintaining the alga at the desired depths generally ensures the light saturated rate of photosynthesis on clear days, and enhances production. Under the cultivated condition, the calculated annual primary productivity of this alga is 160 gC m−2 y−1. Translocation of 14C-labelled photoassimilates in rapidly growing sporophyte of Undaria pinnatifida was studied in the open sea. Samples from different parts of the blade with counterparts exposed to tracer (NaH14CO3) showed that the translocation that occurred mainly from the tip of the blade to the growing region had obvious source-sink relationship. It took 20 minutes to translocate the labelled photoassimilates from the epidermis, via cortex, to the medulla of the midrib, where rates of translocation averaging 42–48 cm h−1 were observed in the open sea. Production experiments of tip-cutting of the blades showed an increased production of 9%.
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  • 86
    ISSN: 1573-5117
    Keywords: stream ; disturbance ; caddisfly ; shredder ; diet ; growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Diet and growth of leaf-shredding caddisfly larvae, Pycnopsyche spp.,were examined in streams draining a reference catchment and a 16-year-oldclear-cut (disturbed) catchment at Coweeta Hydrologic Laboratory insouthwestern North Carolina, USA. The objective was to explain why shredderproduction is higher in the disturbed streams despite the larvae having lessfood (i.e., leaves) available. We predicted larvae would grow faster onfast-decaying leaf material representative of the disturbed streams. Larvaeconsumed mostly leaf detritus in three streams draining each catchment overthree seasons (fall, winter, and spring), which showed larvae did notconsume higher quality foods (e.g., algae and animal material) in disturbedstreams. When fed 2-month-old conditioned black birch (Betula lenta L.) (afast-decaying leaf species) and white oak (Quercus alba L.) (a slow-decayingleaf species) leaves in the laboratory, larvae grew significantly faster onthe birch leaves. However, when larvae were fed the same leaf types after3-months conditioning, larvae grew significantly faster on oak leaves. Afield growth experiment conducted for 42 d using mixed-species leaf dietsrepresentative of each catchment and initially conditioned for 2 monthsfound that Pycnopsyche grew significantly better on the diet representativeof the reference catchment. The ’reference diet‘ contained more oak leaveswhich apparently became a more acceptable food as the experiment proceeded.High shredder production in the disturbed streams could not be explained byhigh Pycnopsyche growth rates on fast-decaying leaves. Instead, larvae grewbetter on leaves that were apparently conditioned optimally regardless ofconditioning rate.
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  • 87
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    Hydrobiologia 351 (1997), S. 35-40 
    ISSN: 1573-5117
    Keywords: flora ; endemic plant ; biogeography ; conservation ; Greece
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The flora of Prespa National Park is composed of 1326plant species. Among them are 17 Pteridophytes and1309 Spermatophytes, represented by 11 Gymnosperms,1057 Dicotyledons and 241 Monocotyledons. The flora isevaluated and categorised into species which can beconsidered as new for the Greek flora, species underinternational protection, and rare local speciesthreatened with extinction. Plant species are alsocategorised according to their ecologicalrequirements, with special reference to the endemicspecies and their centres of origin in respect totheir total percentage (15.4%).
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  • 88
    ISSN: 1573-5117
    Keywords: grouper larvae ; live foods ; growth ; fatty acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fertilized oyster eggs, S-rotifer (Brachionusrotundiformis) and SS-rotifer (Brachionus sp.),were tested either solely or in combination fortheir suitability as feed for early stage grouper (Epinephelus coioides) larvae. Sizes ofS-rotifers ranged between 143-224 µm lorica lengthwith mean 182±21µm, and SS-rotifersbetween 122-176µmlorica length with mean 154±13µm. Theresults indicated that both fertilized oyster eggs andSS-rotifers were suitable as feed at F1 (first feedingday). However, poor growth was recorded when providing oyster eggs solely for the periodsF1-F3. Although growth of larvae at F6 had nodifference between the sole SS-rotifers and the oystereggs additionally provided for F1-F3, better survivalof larvae at F15 was obtained when providingcombinations of SS-rotifers with oyster eggs for F1-F3.Besides, better growth and survival of larvae at F15was found when providing S-rotifers enriched withKirin yeast for F7-F15. The highest survival andfastest growth of larvae at F15 was found whenproviding oyster eggs for F1-F3, SS-rotifers forF1-F6, S-rotifers for F7-F15, both rotifers enrichedwith Kirin yeast, and Isochrysis for F1-F15.Total fatty acid (TFA), EPA, DHA content, and DHA/EPA(D/E) ratio of larvae changed with their sizes andcorresponded to that of their feeds. The F15 larvaehaving a higher TFA grew faster, having higher DHA orEPA survived better.
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  • 89
    ISSN: 1573-5028
    Keywords: elongation ; gene expression ; glucanase ; Arabidopsis thaliana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The isolation of an elongation-specific endo-1,4-β-glucanase-cel1 from Arabidopsis thaliana was made possible by the fact that considerable homology exists between different endo-1,4-β-glucanase (EGase) genes from different plants. Degenerate primers were synthesized based on two conserved regions from the avocado and tomato cellulase amino acid sequences. The A. thaliana cel1 cDNA gene was found to encode a 54 kDa protein; sequence comparison with the avocado EGase revealed 56% identity. Northern blot analysis of cel1 suggested its developmental regulation. RNA transcripts were undetectable in fully expanded leaves as well as at the basal internode of flowering stems. However, a strong transcript signal was detected in the elongating zone of flowering stems of normal plants. The RNA transcript level of cel1 in the elongating zone of dwarf flowering stems was significantly lower than in the corresponding zone in normal plants. This suggests cel1's involvement in cell elongation in A. thaliana. Transgenic tobacco plants transformed with the putative cel1 promoter region fused to the gus reporter gene, showed a significant GUS staining both in shoot and root elongating zones. These results further substantiate the link between cel1 expression and plant cell elongation.
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  • 90
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    Plant molecular biology 35 (1997), S. 69-77 
    ISSN: 1573-5028
    Keywords: rice ; wild rice ; Oryza spp. ; evolution ; conservation ; evaluation ; utilization ; germplasm ; genetic resources ; genebank
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rice has been grown in Japan for about 3000 years. Although both japonica and indica varieties have been grown in Japan, now japonica rices are grown. Japanese rice breeding has used an ecological breeding approach. While emphasis in rice breeding in the 1940's and 1950's focussed on yield in recent decades quality has been of major importance. Consumer preference and name recognition of high quality varieties, such as Koshihikari, has resulted in slow acceptance of new varieties. Rice germplasm was systematically collected throughout Japan between 1962 and 1963. Subsequent acquisition and collecting, in Japan and other countries, has resulted in 28,000 accessions being conserved in the National Genebank, based at the National institute of Agrobiological Resources (NIAR). Research on genetic diversity of rice using a range of techniques, for example esterase isozymes, has revealed clinal variation in rice radiating from the center of diversity of rice in and around southwest China. Newly found genes in traditional rice germplasm, such as genes for non-elongating mesocotyl, are now routinely identified on the rice genome. Pioneering studies on eco-genetic differentiation of species in the genus Oryza in Japan has revealed much about the complex genepool for which rice evolved. Pest and disease resistance sources, particularly to blast, bacterial blight and brown plant hopper, from many countries have been incorporated into Japanese varieties. Cold tolerance at the booting stage was found in the Indonesian variety Silewah. In the future in characterisation of rice germplasm and interaction between rice germplasm specialists and rice molecular scientists, both in Japan and internationally, will be corner stones to securing rice genetic diversity and rice improvement in the next century.
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  • 91
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    Plant molecular biology 35 (1997), S. 293-302 
    ISSN: 1573-5028
    Keywords: Arabidopsis ; blue light ; gene expression ; Lhcb ; transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Irradiation of etiolated Arabidopsis or pea, or dim-red-light-grown pea seedlings with a single, short (under 10 s) pulse of blue light (threshold at 0.1 µmol/m2) is sufficient to induce the expression of specific members of the Lhcb gene family including the pea Lhcb1*4 gene and the Arabidopsis Lhcb1*3 gene. Other Lhcb genes, such as the pea Lhcb1*3 gene and the Arabidopsis Lhcb1*1 and 1*2 genes are unaffected by this blue-light treatment. Transgenic Arabidopsis bearing pea Lhcb1*3::Gus (β-glucuronidase), pea Lhcb1*4::Gus or Arabidopsis Lhcb1*3::Gus constructs were used to determine if pea and Arabidopsis employ a similar mechanism to achieve blue-light induced Lhcb expression. Examination of the respective Gus expression patterns in white-light-grown seedlings indicates that the pea promoters are active and properly expressed in the Arabidopsis background. Irradiation of dark-grown Arabidopsis with a 20 s pulse of blue light with a total fluence of 100 µmol/m-2 results in expression of the pea Lhcb1*4::Gus (β-glucuronidase) construct, but not of the pea Lhcb1*3::Gus construct indicating that the pea promoters respond correctly to blue light in the Arabidopsis background. Fluence-response, time-course and reciprocity characteristics for the blue-light-induced expression of the pea Lhcb1*4::Gus construct closely resemble those of the endogenous Arabidopsis Lhcb genes, confirming the proper interpretation of the Arabidopsis blue-light-signaling mechanism by the pea Lhcb1*4 promoter and suggesting that the signaling mechanisms in the two plants are very similar, if not identical. Fluence response data for the steady-state level of transcript derived from an Arabidopsis Lhcb1*3::Gus construct extending 200 bp upstream of the site of transcription indicate that the blue light responsive element(s) are contained within this 200 bp region.
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  • 92
    ISSN: 1573-5028
    Keywords: Betula pendula Roth ; differential display ; gene expression ; mitochondria ; oxidative stress ; ozone ; phosphate translocator
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated by DDRT-PCR (differential-display reverse-transcription polymerase chain reaction) and cDNA library screening a 1.3 kb cDNA corresponding to a strongly ozone-inducible transcript from birch (Betula pendula Roth). Nucleotide sequence analysis suggests that it encodes a mitochondrial phosphate translocator protein (Pi c), the first one isolated from plants. The isolated birch mitochondrial phosphate translocator cDNA (designated Mpt1) contains an open reading frame of 1092 bases encoding a 364 amino acid polypeptide. The deduced protein is 66% similar to bovine Pic isoform B. Comparison of the N-terminal amino acid sequence to known mammalian Pic proteins and the existence of an in-frame stop codon upstream of the initiation codon suggest that the isolated cDNA is full-length. Southern hybridization analysis of birch genomic DNA shows that Mpt1 is a single-copy gene. Accumulation of Mpt1 mRNA during oxidative stress imposed by ozone is detectable already at 2 h and it is at maximum ca. 12 h after the beginning of an 8 h ozone exposure (150 ppb). A second O3 peak at 48–56 h did not increase transcript levels further. O3 exposure for 2 h was sufficient for Mpt1 induction. Birch Mpt1 transcript levels remain at moderately low level during leaf development and is lower in roots and leaves when compared to young shoots undergoing wood formation and lignification.
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  • 93
    ISSN: 1573-5028
    Keywords: competitive PCR ; flavonoid pathway ; Forsythia ; gene expression ; transformation ; woody ornamentals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression, during flower development, of the gene encoding the anthocyanin pathway key enzyme dihydroflavonol 4-reductase (DFR) was investigated in floral organs of Forsythia × intermedia cv. ‘Spring Glory’. Full-length DFR and partial chalcone synthase (CHS) cDNAs, the gene of interest and a flavonoid pathway control gene respectively, were obtained from petal RNA by reverse transcription PCR. Whereas for CHS northern blot analysis enabled the study of its expression pattern, competitive PCR assays were necessary to quantify DFR mRNA levels in wild-type plants and in petals of 2 transgenic clones containing a CaMV 35S promoter-driven DFR gene of Antirrhinum majus. Results indicated a peak of CHS and DFR transcript levels in petals at the very early stages of anthesis, and different expression patterns in anthers and sepals. In comparison to wild-type plants, transformants showed a more intense anthocyanin pigmentation of some vegetative organs, and a dramatic increase in DFR transcript concentration and enzymatic activity in petals. However, petals of transformed plants did not accumulate any anthocyanins. These results indicate that other genes and/or regulatory factors should be considered responsible for the lack of anthocyanin production in Forsythia petals.
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  • 94
    ISSN: 1573-5028
    Keywords: cortex ; gene expression ; in situ hybridization ; organ-preferential ; root ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A near full-length cDNA clone (pZRP2) was isolated from a cDNA library constructed from maize root mRNAs. The predicted polypeptide has a calculated molecular mass of 66 975 Da, is largely hydrophilic, and contains 26 repeats of a motif the consensus sequence of which is RKATTSYG[S][D/E][D/E][D/E][D/E][P]. The function of the putative protein remains to be elucidated. The ZRP2 mRNA accumulates to the highest levels in young roots, and is also present in mature roots and stems of maize. Further analysis of young roots indicates that the lowest level of ZRP2 mRNA is near the root tip, with relatively high levels throughout the remainder of the root. In situ hybridization reveals that ZRP2 mRNA accumulates predominantely in the cortical parenchyma cells of the root. In vitro nuclear run-on transcription experiments indicate a dramatically higher level of zrp2 gene transcription in 3-day old roots than in 5-day old leaves. A zrp2 genomic clone, which includes the transcribed region and 4.7 kb of upstream sequence, was isolated and characterized.
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  • 95
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    Plant molecular biology 35 (1997), S. 433-442 
    ISSN: 1573-5028
    Keywords: chickpea ; epicotyls ; growth ; tissue-specific gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two cDNAs, CanST-1 and CanST-2, encoding two different growth-related organ-specific sequences have been isolated from a cDNA library from growing epicotyls of Cicer arietinum. An intriguing property of these two clones is the presence in their coding region of a repeated sequence which is highly conserved except for the number of repeats. The corresponding genes of CanST-1 and CanST-2 encode for proteins related to elongation processes. CanST-1 and CanST-2 are up-regulated during epicotyl growth, the transcript levels of both clones decrease when the growth of epicotyls is inhibited by several treatments and their expression increases when epicotyls resume growth. Furthermore, clones CanST-1 and CanST-2 are tissue-specific and are only expressed in epicotyls, mesocotyls, roots and stem tissues whose cells undergo elongation processes. Neither clone was found to be expressed in other organs such as cotyledons, leaves, flowers, pods and immature seeds. The results of auxin (IAA) and brassinolides (BR) treatments suggest that the processes in which the proteins encoded by CanST-1 and CanST-2 are involved are not mediated by these hormones.
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  • 96
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    International journal of primatology 18 (1997), S. 683-701 
    ISSN: 1573-8604
    Keywords: capuchins ; chimpanzees ; growth ; reproduction ; weaning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We examined growth and development in capuchins and chimpanzees in relation to weaning, onset of reproduction, and reproductive life span. Striking differences are evident in neurobehavioral status at birth (more mature in capuchins), the relative duration of infancy (longer in chimpanzees), and the proportional weight of the infant at the time of weaning (greater in capuchins). Although capuchins and chimpanzees spend a similar proportion of life in a weaned but reproductively immature state, chimpanzees spend so much more of their lives as nursing infants that reproductive output per individual is much lower than in capuchins. Discussion centers around tolerated transfers of food (food-sharing) as a potential adaptation to limited foraging success by immature foragers. Perhaps food transfers from adult to infant, which is a more prominent feature of behavior in chimpanzees than in capuchins in natural environments, allow a very small weanling chimpanzee to survive.
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  • 97
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    International journal of primatology 18 (1997), S. 995-1004 
    ISSN: 1573-8604
    Keywords: De Brazza's monkey ; Cercopithecus neglectus ; survey ; conservation ; Kenya
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We conducted a trail survey of De Brazza's monkey (Cercopithecus neglectus) on Mt. Elgon and the Cherangani Hills between October 1994 and February 1995. The objectives were to assess the status and distribution of the species and its habitats and to formulate recommendations on its conservation. We counted a total of 49 monkeys: 3 groups and 6 lone individuals near Kimothon River (Mt. Elgon) and 4 groups and 6 lone individuals at Kapolet Forest Reserve and its environs (Cherangani Hills). The mean group size is 6.6. Wanton habitat destruction was evident in all the forest habitats. The species now inhabits unprotected remnant strips of riverine forest. Furthermore, the Kapolet Forest Reserve offers little or no protection to De Brazza's monkeys or their habitat. Translocation of the monkeys from unprotected areas to a protected habitat is recommended as an urgent conservation measure to save the De Brazza's population in Mt. Elgon and the Cherangani areas of Kenya.
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  • 98
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    Wetlands ecology and management 5 (1997), S. 19-36 
    ISSN: 1572-9834
    Keywords: wetlands ; rehabilitation ; conservation ; Australia ; exotic species
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The wetlands of the Alligator Rivers Region of northern Australia have been recognized as having high national and international conservation value. The diversity and productivity of these habitats is, however, under current and future threat from invasive feral animals (Asian water buffalo and pigs) and naturalized alien plants (mimosa, salvinia and para grass) and also from climate change and sea level rise. Some habitats have already been severely degraded and require rehabilitation. In response to this situation feral animal management has centered on eradicating the buffalo population and introducing measures to control pigs. Weed management has focused on control of mimosa and salvinia with an increasing emphasis on integrated control measures. The vulnerability of the freshwater wetlands to climate change and sea level rise is considerable, although the exact nature of environmental change has not been determined. Rehabilitation of the degraded wetlands has centered on controlling the cause of the problem (e.g., the invasive species) and allowing subsequent natural succession to occur. It is recommended that further monitoring and assessment of successional change is undertaken to ascertain if this is sufficient. An integrated monitoring program for detecting the extent and rate of ecological change because of climate change and sea level rise is also proposed. Specific management and research tasks for each of the major broad causes of wetland degradation within the region are made. It is then strongly recommended that all rehabilitation and monitoring activities, including related research, are conducted within a holistic management framework that takes into account the different land jurisdictions within the region and also within the broader landscape context. The utilization of existing management and research structures and processes is stressed as one means of achieving an integrated approach.
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  • 99
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    Wetlands ecology and management 5 (1997), S. 215-225 
    ISSN: 1572-9834
    Keywords: floating weeds ; effects ; wetlands ; conservation ; control ; management
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The most important floating aquatic weeds (FAWs) are Eichhornia crassipes, Salvinia molesta and Pistia stratiotes. E. crassipes and P. stratiotes reproduce sexually. All three species reproduce asexually. E. crassipes and S. molesta have particularly high growth rates. All can form dense mats and growth rates are increased by high nutrient levels and temperatures. Spread between continents and watersheds is largely the result of human activities. Spread within watersheds is mostly via floating propagules. FAWs are known to affect water resource management, the continued existence of human riverine and wetland communities, and conservation of biodiversity. Waterways can be blocked, and the efficiency of irrigation and hydro generation impaired. People are affected by reduction of the fish catch, inability to travel by boat and consequent isolation from gardens, markets and health services, and also changes in populations of vectors of human and animal diseases. Biodiversity can be reduced and conservation value affected. It is proposed that rational application of physical, chemical and biological control of FAWs, and reduction of nutrient input should be part of every strategy for the sustainable management of wetlands.
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  • 100
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    Pharmaceutical research 14 (1997), S. 1122-1126 
    ISSN: 1573-904X
    Keywords: dissolution ; model ; growth ; fraction absorbed ; in vitro-in vivocorrelations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. To develop a new approach for describing drug dissolution which does not require the presuppositions of time continuity and Fick's law of diffusion and which can be applied to both homogeneous and heterogeneous media. Methods. The mass dissolved is considered to be a function of a discrete time index specifying successive 'generations' (n). The recurrence equation: Φ n+1 = Φ n + r(l − Φn)(1 − Φn X 0/θ) was derived for the fractions of dose dissolved Φ n and Φn +1, between generations n and n + 1, where r is a dimensionless proportionality constant, X 0 is the dose and θ is the amount of drug corresponding to the drug's solubility in the dissolution medium. Results. The equation has two steady state solutions, Φ ss = 1 when (X 0/θ) ≤ 1 and Φ ss = θ/X 0 when (X 0/θ) 〉 1 and the usual behavior encountered in dissolution studies, i.e, a monotonic exponential increase of Φ n reaching asymptotically the steady state when either r 〈 θ/X 0 〈 1 or r 〈 1 〈 θ/X 0. Good fits were obtained when the model equation was applied to danazol data after appropriate transformation of the time scale to 'generations'. The dissolution process is controlled by the two dimensionless parameters θ/X 0 and r, which were found to be analogous to the fundamental parameters dose anddissolution number, respectively. The model was also used for the prediction of fraction of dose absorbed for highly permeable drugs. Conclusions. The model does not rely on diffusion principles and therefore it can be applied under both homogeneous and non-homogeneous conditions. This feature will facilitate the correlation of in vitro dissolution data obtained under homogeneous conditions and in vivo observations adhering to the heterogeneous milieu of the GI tract.
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