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    Electronic Resource
    Electronic Resource
    Selektive Partnerwahl der Aaskrähe (Corvus corone) in der Hybridisierungszone von Rabenkrähe (C. c. corone) und Nebelkrähe (C. c. cornix) (1998)
    Springer
    Journal of ornithology 139 (1998), S. 173-177 
    Details
    ISSN: 1439-0361
    Keywords: mate choice ; taxonomy ; phenotypic hybrids ; fitness ; decision rule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Zusammenfassung Die als Unterarten klassifizierten europäischen Formen der Aaskrähe, Rabenkrähe und Nebelkrähe, besiedeln verschiedene, aneinandergrenzende Verbreitungsgebiete und hybridisieren in der Kontaktzone. Die Nachkommen von Mischpaaren sind fruchtbar und können sowohl mit anderen Hybriden als auch mit Raben- und Nebelkrähen erfolgreich brüten. Trotzdem kommt es zu keiner völligen Vermischung der Formen und/oder Verlagerung der Verbreitungsgebiete. Vor diesem Hintergrund untersuchten wir die Partnerwahl von Aaskrähen in der Hybridisierungszone auf der nordfriesischen Insel Amrum und stellten fest, daß Partner gleichen Phänotyps häufiger miteinander verpaart waren, als stochastisch zu erwarten gewesen wäre. Unsere Daten bestätigen vergleichbare Studien aus Hybridisierungszonen in Italien und Sibirien. Wir schließen daraus, daß phänotypisch selektive Partnerwahl bei der Aaskrähe ein allgemeines Phänomen sein könnte und diskutieren, warum dieses Verhalten anfitness-relevante Parameter gekoppelt sein sollte. Um welche es sich dabei handeln könnte, wurde bisher nicht hinreichend untersucht und muß deshalb offen bleiben.
    Notes: Summary Carrion Crow and Hooded Crow are regarded as subspecies of the Crow. They show frequent hybridisation along the adjacent borders of their distribution. Mixed pairs produce fertile offspring which are able to breed successfully with both hybrids and mates of either phenotype. However, hybridisation does not lead to phenotypic changes of Carrion and Hooded Crows in general nor in their distinct distribution. We studied the mating behaviour of Crows in the hybrid zone on the Island of Amrum (Schleswig-Holstein, Germany) and found evidence that Crows may prefer mates of the same phenotype. Our data confirm previous studies which reported assortative mating with respect to plumage coloration from hybrid zones in Italy and Siberia. We discuss why this behaviour should be related tofitness traits which in our opinion have not yet been studied adequately nor identified.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01651226
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  • 2
    Electronic Resource
    Electronic Resource
    Influence of cereal leaf epicuticular wax on Diuraphis noxia probing behavior and nymphoposition (1998)
    Springer
    Entomologia experimentalis et applicata 89 (1998), S. 111-118 
    Details
    ISSN: 1570-7458
    Keywords: leaf surface wax ; probing behavior ; nymphoposition ; Russian wheat aphid ; wheat ; barley ; oat ; Homoptera ; Aphididae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of cereal leaf surface wax on Diuraphis noxia (Mordvilko), the Russian wheat aphid, probing behavior and nymphoposition was evaluated. Ultrastructure of leaf epicuticular wax from wheat (Triticum aestivum L.) c.v. ‘Arapahoe’ and ‘Halt’ was different from barley (Hordeum vulgare L.) c.v. ‘Morex’, and oat (Avena sativa L.) c.v. ‘Border’. Both wheat cultivars had similar rod-shaped epicuticular wax, while barley and oat plants had flakes. The chemical composition comparison of gas chromatograms also indicated that the extract of the two wheat cultivars had similar pattern of peaks, while the barley and oat leaves had similar peaks. Cereal variety significantly affected aphid probing behavior (P 〈 0.05), but wax removal using ethyl ether swab did not (P 〈 0.05). Aphids initiated significantly more probes on Border oat leaves than on Morex barley irrespective of wax removal, although total probing duration per aphid was not significantly different among the four cereals examined. Accumulative salivation duration per aphid on oat leaves with wax was significantly longer than other cereal leaves with wax, while accumulative ingestion duration per aphid on Arapahoe wheat and Morex barley was significantly longer than on oat. Nymphoposition of D. noxia on cereal leaves maintained on the benzimidazole-agar medium showed that aphids produced a greater number of nymphs on Morex barley and less on Border oat leaves, although wax removal did not affect aphid nymphoposition. Removal of leaf epicuticular waxes from the 4 cereal genotypes using ethyl ether swab indicated that the influence of wax on plant resistance to D. noxia probing and reproduction was limited. Morex barley was the most favorable, while Border oat was the least favorable cereal host of D. noxia.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003458406502
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  • 3
    Electronic Resource
    Electronic Resource
    Nuclear calcium: a key regulator of gene expression (1998)
    Springer
    BioMetals 11 (1998), S. 345-358 
    Details
    ISSN: 1572-8773
    Keywords: calcium ; CREB ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Through the evolution of multicellular organisms, calcium has emerged as the preferred ion for intracel-lular signalling. It now occupies a pivotal role in many cell types and nowhere is it more important than in neurons, where it mediates both the relaying and long-term storage of information. The latter is a process that enables learning and memory to be formed and requires the activation of gene expression by calcium signals. Evidence from a number of diverse organisms shows that transcription mediated by the transcrip-tion factor CREB is critical for learning and memory. Here we review the features of CREB activation by calcium signals in mammalian cells. In contrast to other transcription factors, its regulation is dependent on an elevation of nuclear calcium concentration, potentially placing this spatially distinct pool of calcium as an important mediator of information storage.© Kluwer Academic Publishers
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009257909785
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  • 4
    Electronic Resource
    Electronic Resource
    Multiplex Titration RT-PCR: Rapid Determination of Gene Expression Patterns for a Large Number of Genes (1998)
    Springer
    Plant molecular biology reporter 16 (1998), S. 323-339 
    Details
    ISSN: 1572-9818
    Keywords: Aux/IAA genes ; gene expression ; gene families ; RT-PCR ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have developed an improved method for determination of gene expression levels with RT-PCR. The procedure is rapid and does not require extensive optimization or densitometric analysis. Since the detection of individual transcripts is PCR-based, small amounts of tissue samples are sufficient for the analysis of expression patterns in large gene families. Using this method, we were able to rapidly screen nine members of the Aux/IAA family of auxin- responsive genes and identify those genes which vary in message abundance in a tissue- and light-specific manner. While not offering the accuracy of conventional semi-quantitative or competitive RT-PCR, our method allows quick screening of large numbers of genes in a wide range of RNA samples with just a thermal cycler and standard gel analysis equipment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007523305132
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  • 5
    Electronic Resource
    Electronic Resource
    Identity of Fusarium nygamai isolates with long and short microconidial chains from millet, sorghum and soil in Africa (1997)
    Springer
    Mycopathologia 140 (1997), S. 171-176 
    Details
    ISSN: 1573-0832
    Keywords: Africa ; Fusarium ; F. moniliforme ; grain ; Lesotho ; mating population ; Nigeria ; taxonomy ; Zimbabwe
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Several Fusarium species have been found associated with millet and sorghum in Nigeria, Lesotho and Zimbabwe. Amongst these, some isolates were originally identified as short- and long-chained types of F. nygamai. However, there was some question as to the correct identification of the long chained types. This study reclassified some of the isolates with long microconidial chains as F. moniliforme. Morphologically, these strains do not produce chlamydospores like F. nygamai, but produce swollen hyphal cells or resistant hyphae. The isolates in this study were crossed with the mating-type tester strains of Gibberella fujikuroi (F. moniliforme and G. nygamai (F. nygamai). Of the isolates with long chains of microconidia and other characteristics of F. moniliforme, 36% crossed with mating population ''A'' of G. fujikuroi. Of the isolates with characteristics of F. nygamai, 65% crossed with the testers used to produce the teleomorph of F. nygamai. Mating tests support the separation of the sample population into F. moniliforme and F. nygamai. The results of this study show that genetics can be an aid in resolving some problems in fungal taxonomy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006863825469
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  • 6
    Electronic Resource
    Electronic Resource
    A survey on the occurrence of mycotoxins in wheat and maize from western Romania (1998)
    Springer
    Mycopathologia 143 (1998), S. 97-103 
    Details
    ISSN: 1573-0832
    Keywords: deoxynivalenol ; enzyme immunoassay ; feed ; maize ; mycotoxins ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Samples of wheat (n = 25) and maize (n = 30) for animal consumption, collected in 1997 after harvest from western Romania, were analyzed by enzyme immunoassays for mycotoxin contamination. Toxins analyses included deoxynivalenol (DON), 3-acetylDON, 15- acetylDON, fusarenone X (FX), T-2 Toxin (T-2), diacetoxyscirpenol (DAS), zearalenone (ZEA), fumonisin B1 (FB1), aflatoxin B1 (AFB1), ochratoxin A (OA), and citrinin (CT). DON and acetylDONs were the major contaminants in wheat (100%) and maize (46%). Median values for DON, 3-acetylDON, and 15-acetylDON were 880 μg kg-1, 66 μg kg- 1, and 150 μg kg-1 in wheat, and 890 μg kg-1, 180 μg kg-1, and 620 μg kg- 1 in maize, respectively. Additionally, 3,15-diacetylDON was detected in some samples by HPLC-EIA analysis. All samples were negative for FX (〈150 μg kg-1). T-2 was found in wheat (n = 6) and maize (n = 1) at levels between 13 and 63 μg kg- 1. DAS (2.6 μg kg-1) was found in one maize sample. ZEA occurred in all wheat and in four maize samples, median values were 10 μg kg-1 and 250 μg kg-1, respectively. One maize sample contained FB1 (140 μg kg-1). All samples were AFB1-negative (〈4 μg kg-1). OA was found in one wheat sample (37 μg kg- 1), CT was found in one maize sample (580 μg kg- 1). This first reported natural occurrence of a range of mycotoxins in Romanian feeding stuff shows that DON and acetyl DONs may be present at levels which may affect animal production.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006987205986
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  • 7
    Electronic Resource
    Electronic Resource
    Candida novakii, sp. nov. a new anamorphic yeast species of ascomycetous affinity (1997)
    Springer
    Antonie van Leeuwenhoek 71 (1997), S. 375-378 
    Details
    ISSN: 1572-9699
    Keywords: Candida novakii ; taxonomy ; yeasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two strains of an undescribed species of the genus Candida were isolated from decaying wood of Quercus sp. A description of the new species Candida novakii is given.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1000238205640
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  • 8
    Electronic Resource
    Electronic Resource
    Verrucomicrobia div. nov., a new division of the Bacteria containing three new species of Prosthecobacter (1997)
    Springer
    Antonie van Leeuwenhoek 72 (1997), S. 29-38 
    Details
    ISSN: 1572-9699
    Keywords: phylogeny ; prosthecobacter ; taxonomy ; Verrucomicrobia ; Verrucomicrobiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four strains of nonmotile, prosthecate bacteria were isolated in the 1970s and assigned to the genus Prosthecobacter. These strains were compared genotypically by DNA/DNA reassociation and 16S rDNA based phylogenetic analyses. Genotypic comparisons were complemented with phenotypic characterizations. Together, these studies clearly indicate each Prosthecobacter strain represents a novel species of bacteria. We propose three new species of Prosthecobacter, P. dejongeii strain FC1, P. vanneervenii strain FC2, and P. debontii strain FC3; P. fusiformis is reserved for the type strain of the genus, strain FC4. Additionally, we propose the genera Prosthecobacter and Verrucomicrobium, currently members of the order Verrucomicrobiales, to comprise a novel higher order taxonomic group, the division Verrucomicrobia div. nov. and the class Verrumicrobiae class nov. Many novel members of the Verrucomicrobia, as revealed by molecular ecology studies, await isolation and description.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1000348616863
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  • 9
    Electronic Resource
    Electronic Resource
    Studies on keratinophilic fungi. Ix: Neoarachnotheca gen. nov. and a new species of Nannizziopsis (1997)
    Springer
    Antonie van Leeuwenhoek 72 (1997), S. 149-158 
    Details
    ISSN: 1572-9699
    Keywords: keratinophilic fungi ; Neoarachnotheca ; Neoarachnotheca keratinophila ; Nannizziopsis tropicalis ; Onygenales ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Neoarachnotheca is proposed as a new genus of Onygenales. The outstanding generic characteristics are white, spherical ascomata with a wall formed by a network of hyphae and spherical, subhyaline ascospores with an irregular sheath. Nt. keratinophila, the type species, characterized by wavy peridial hyphae has been isolated from marine and river sediments and Myriodontium keratinophilum is its anamorph. Nannizziopsis tropicalis is proposed as a new species based on a strain isolated from soil in Burundi. RFLPs analysis of ITS and 5.8S rDNA support these proposals. The differences with related genera are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1000394118040
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  • 10
    Electronic Resource
    Electronic Resource
    Use of a lux-based procedure to rapidly visualize root colonisation by Pseudomonas fluorescens in the wheat rhizosphere (1997)
    Springer
    Antonie van Leeuwenhoek 72 (1997), S. 365-372 
    Details
    ISSN: 1572-9699
    Keywords: bioluminescence ; Pseudomonas ; root colonization ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The bioluminescently marked Pseudomonas fluorescens strain 5RL, has been used previously to follow colonisation of soy bean roots (De Weger et al. [1991] Appl. Environ. Microbiol. 57:36-41). In the present paper the method has been further developed and optimized for wheat roots and it is used to get a quick overview of the colonisation patterns of many different root systems at the same time. Colonisation was followed on wheat plants grown in our gnotobiotic sand system (Simons et al., 1996. Mol Plant Microbe Interact 9: 600–607) and the following results were obtained. (i) A spatio-temporal analysis of the colonisation of wheat roots showed that 4 days after planting the highest bacterial activity was observed at the upper part of the root. After 6 days the high bacterial activity at the upper part was further increased, whereas spot-like activities were observed on the lower root parts, possibly due to micro-colonies. (ii) Bacterial mutations causing lack of motility or auxotrophy for amino acids resulted in impaired colonisation of the lower root parts, indicating that motility and prototrophy for the involved amino acid(s) are important factors for wheat root colonisation by strain 5RL. (iii) Coinoculation of strain 5RL with other wild type Pseudomonas strains on the root influenced the colonisation pattern observed for strain 5RL. Colonisation was not visually affected when the competing strain was a poor root coloniser, but was severely reduced when the competing strain was a good root coloniser. The results show that the spatio-temporal colonisation of wheat root by P. fluorescens strain 5RL and derivatives is similar to that of strain WCS365 on tomato. The advantage of the use of lux-marked strains is that the results are obtained much quicker than when conventional methods are used and that the result is supplied as an image of the colonisation pattern of many different roots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1000565413024
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  • 11
    Electronic Resource
    Electronic Resource
    Dipodascus capitatus, Dipodascus spicifer and Geotrichum clavatum: Genomic characterization (1998)
    Springer
    Antonie van Leeuwenhoek 74 (1998), S. 229-235 
    Details
    ISSN: 1572-9699
    Keywords: Dipodascus capitatus ; D.spicifer ; Geotrichum clavatum ; yeast ; taxonomy ; DNA heterogeneity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The G+C contents of 25 strains of Dipodascus capitatus, Dipodascus spicifer and Geotrichum clavatum were found to be heterogeneous on basis of derivative graphs of the melting profiles. Strains showing similar derivative graphs of the melting curve exhibited high levels of DNA homology (80-100%); strains showing dissimilar derivative graphs exhibited low levels of DNA homology (5 to 45%). Being considered separate taxa on basis of these parameters, D. capitatus, D. spicifer and G. clavatum could be identified by a combination of the key characteristics growth on xylose, cellobiose, salicin and arbutin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1001762024710
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  • 12
    Electronic Resource
    Electronic Resource
    Identification of Taxa in the Genus shape Beta using ITS1 Sequence Information (1998)
    Springer
    Plant molecular biology reporter 16 (1998), S. 147-155 
    Details
    ISSN: 1572-9818
    Keywords: allele-specific PCR ; Beta ; ITS1 ; plant identification ; rDNA ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sequence variation in the ITS1 locus of the nuclear ribosomal DNA in beets has previously been used to reconstruct phylogeny of the species in the genus Beta. We have developed protocols that allow the identification of Beta taxa by use of taxon-specific primers. Beta sections, species and subspecies can be identified. Differences within the ITS1 region of a single base can be exploited for species identification. The results from this study not only provide effective methods for wild beet identification, but also indicate the potential use of the techniques in other crops.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007416817736
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  • 13
    Electronic Resource
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    The taxonomic impediment in orthopteran research and conservation (1998)
    Springer
    Journal of insect conservation 2 (1998), S. 151-159 
    Details
    ISSN: 1572-9753
    Keywords: Orthoptera ; biodiversity ; taxonomy ; conservation.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Nature of Science, Research, Systems of Higher Education, Museum Science
    Notes: Abstract It is estimated that only 10–15% of the world's insect fauna has been described and named. Efforts to inventory insect biodiversity are hampered by this taxonomic impediment, which is compounded by the logistical problems of an insufficient taxonomic workforce and their remote location in museums thousands of miles from the areas of highest biodiversity. Compared to most other invertebrate groups however, the taxonomic impediment is relatively benign in the order Orthoptera. This is a small to medium-sized order (approximately 20 000 described species) which is well known taxonomically, owing to the group's agricultural importance worldwide. Furthermore, orthopteran taxonomists are now fortunate to have a published up-to-date catalogue of all known species, which has just become accessible as a regularly updated database on the World Wide Web. Whilst new information technology, in the form of e-mail networks, World Wide Web sites and CD-ROM information archives, is already enhancing communication between specialists and helping to reduce the logistical problems of documenting orthopteran biodiversity, a major reinvestment in basic taxonomic research is needed if we are to reduce the existing taxonomic impediment significantly. There is general agreement that an internationally coordinated approach will be necessary and priorities must be set to tackle the biodiversity/systematics crisis. In the future, the Orthoptera can make an important contribution to invertebrate faunal surveys and have potential as an indicator taxon. Furthermore, the Orthoptera Species File establishes a taxonomic framework which could be readily enlarged to include geographic data and phenology of species from existing museum specimens.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009633811789
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  • 14
    Electronic Resource
    Electronic Resource
    Biological control of pigweeds (Amaranthus retroflexus, L. A. Powellii, S. Watson and A. bouchonii Thell.) with phytophagous insects, fungal pathogens and crop management (1997)
    Springer
    Integrated pest management reviews 2 (1997), S. 51-59 
    Details
    ISSN: 1572-9745
    Keywords: Biological control ; insects ; pathogens ; germination ; taxonomy ; genetic variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Pigweeds (Amaranthus spp.) are of economic importance worldwide. In Europe, Amaranthus retroflexus is one of the ten weed species of greatest economic importance. It is a serious problem weed in several field crops (e.g. maize), as well as in vegetables, orchards and grape vines. It is an annual spreading by seeds which have a long viabilityand are dispersed principally by wind and water, but also by machinery. There is great variability in seed germination which renders control with post-emergence herbicides difficult. In addition, triazine herbicide-resistant populations occur in ten European countries. The aim of this subproject of COST action 816 is to investigate the possibilities of classical and inundative biological control of Amaranthus spp., to characterize potentialbiological control agents and to develop methods for their integration with current phytosanitary measures in the target crops. The project was initiated with an extended literaturesurvey followed by field surveys for insects and pathogens associated with Amaranthus spp. in several European countries. Promising isolates of fungal pathogens have been tested ondetached leaves and whole plants, and initial studies on the application of pathogens causing damping off in seedlings have been made. Further, the variability of different provenances ofAmaranthus spp. in response to fungal attack has been investigated
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1018480429706
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  • 15
    Electronic Resource
    Electronic Resource
    Influence of an acidic beverage (Coca-Cola) on the absorption of itraconazole (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 235-237 
    Details
    ISSN: 1432-1041
    Keywords: Key words Itraconazole ; Coca Cola; acidic beverage ; absorption ; pharmacokinetics ; drug concentration ; food
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: To evaluate the effectiveness of Coca-Cola in enhancing the absorption of itraconazole. Methods: Eight healthy volunteers were randomized to receive two treatment sequences in a two-way crossover design with a 1-week wash-out period separating each study treatment. Treatment I, the control, consisted of 100 mg itraconazole with 325 ml water. Treatment II was identical to treatment I, except that itraconazole was administered with 325 ml of Coca-Cola (pH 2.5). Results: Serum itraconazole concentrations, after administration with Coca-Cola (treatment II), were higher than after administration with water (treatment I). The mean AUC was 1.12 vs 2.02 μg · h · ml−1, the mean Cmax was 0.14 vs 0.31 μg · ml −1and the mean tmax was 2.56 vs 3.38 h in treatments I and II, respectively. Conclusion: The absorption of itraconazole can be enhanced by Coca-Cola.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002280050280
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  • 16
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    Pharmacokinetics of nicardipine enantiomers in healthy young volunteers (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 289-292 
    Details
    ISSN: 1432-1041
    Keywords: Key words Nicardipine; enantiomers ; healthy volunteers ; pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objectives: The present study was conducted to compare pharmacokinetic behaviors of nicardipine enantiomers given in different doses with different formulations of racemic nicardipine in healthy volunteers. Methods: One or two 20-mg racemic nicardipine tablets, and a 40-mg sustained-release capsule of nicardipine were administered to eight healthy volunteers in a cross-over fashion and pharmacokinetic parameters were evaluated. Enantiomer concentrations were determined by GC-MS combined with chiral stationary phase HPLC. Results and conclusions: Serum concentration of (+)-nicardipine was approximately 2–3 times higher than that of (−)-nicardipine in 20- and 40-mg doses of conventional formulations and a non-linear increase in bioavailability with dose was demonstrated. The value for AUC of (+)-nicardipine was approximately 2.3–2.8 times greater than that of the (−)-nicardipine (P 〈 0.05) when 20 and 40 mg racemic nicardipine were administered in a conventional preparation. Relative bioavailability of the sustained-release preparation vs the conventional preparation was 28% and 44% for (+)- and (−)-nicardipine, respectively, for the 40-mg dose.
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    URL: http://dx.doi.org/10.1007/s002280050292
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  • 17
    Electronic Resource
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    Transdermal nitroglycerin: clinical and pharmacokinetic consequences of renewing the patch and the application site (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 379-381 
    Details
    ISSN: 1432-1041
    Keywords: Key words Nitroglycerin; transdermal nitrate ; pharmacokinetics ; patch renewal ; exercise test
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: We examined whether nitroglycerin (glyceryl trinitrate, GTN) patch treatment for 24 h could induce local cutaneous changes that impaired drug delivery and clinical efficacy. Methods: Twenty angina patients were exercise-tested after 2 and 24 h of treatment and then 2 h after patch renewal. The patch was either renewed on a new skin location or on the previous application site in a randomised, double-blind, cross-over protocol. GTN plasma concentrations and finger plethysmography were obtained before and after each exercise test. Results and conclusions: The clinical efficacy, the effect seen on plethysmography and the GTN plasma concentrations tended to increase after patch renewal, regardless of the application site of the renewed patch. Hence, cutaneous changes of clinical importance could not be demonstrated.
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    URL: http://dx.doi.org/10.1007/s002280050304
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  • 18
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    Pharmacokinetics and pharmacodynamic effects of the angiotensin II antagonist valsartan at steady state in healthy, normotensive subjects (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 441-449 
    Details
    ISSN: 1432-1041
    Keywords: Key words Angiotensin II ; Valsartan; AT1 receptor antagonist ; healthy volunteers ; pharmacokinetics ; renin-angiotensin system ; blood pressure ; passive tilting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: Pharmacokinetics, pharmacodynamic effects and tolerability of 200 mg valsartan, once-daily for 8 days, were investigated in 16 healthy, normotensive volunteers on a normal sodium diet. Methods: This was a double-blind, placebo-controlled, randomized crossover study. Drug concentrations in plasma and urine, angiotensin II (Ang II) concentrations in plasma, systolic (SBP) and diastolic (DBP) blood pressure, heart rate (HR) in the supine position and 3 min after passive head-up tilting, as well as safety parameters (ECG, clinical chemistry and hematology, renal water and electrolyte excretion) were measured over 24 h after the first dose (day 1) and at steady state on day 8. Results: Absorption and distribution of valsartan were rapid (Cmax, 2 h; t½λ1 〈 1 h), followed by a slower terminal elimination phase (t½λ2, 6 h) on days 1 and 8, with little accumulation in plasma (increase of 20% on day 8). Less than 10% of the dose was excreted unchanged in urine. The increase in plasma Ang II (Cmax, 6 h) was significantly enhanced at steady state. Supine SBP and DBP significantly decreased on day 8 only, by an average of −3.6 and −2.4 mmHg, respectively, versus placebo, without a concomitant increase in HR. Upon passive tilting, the increase in DBP, normally reinforced by sympathetic renin release, was slightly but significantly blunted on day 1 (−2.0 mmHg) and day 8 (−4.0 mmHg) of treatment with valsartan versus placebo. The orthostatic reflex increase in HR was slightly enhanced compared with placebo by an average of 2.8 beats · min−1 on day 1 and by 2.9 beats · min−1 on day 8. Valsartan was well tolerated and had no influence on ECG, clinical laboratory parameters, and water, electrolyte and uric acid excretion. Conclusions: Pharmacokinetics of valsartan are unchanged after multiple once-daily dosing, with little (expected) accumulation in plasma. Effects of 200 mg valsartan on blood pressure in healthy subjects on a normal sodium intake are small and become more prominent after repeated dosing. Indirect evidence of AT1 blockade by valsartan is demonstrated by an increase of plasma Ang II and by a blunted DBP response to passive tilting. The decrease in blood pressure at steady state enhances the increase in plasma Ang II. Valsartan is well tolerated and is devoid of effects on water, electrolyte and uric acid excretion at 200 mg per day in healthy normotensive volunteers.
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    Cardiovascular effects of different infusion rates of sufentanil in patients undergoing coronary surgery (1997)
    Springer
    European journal of clinical pharmacology 51 (1997), S. 359-366 
    Details
    ISSN: 1432-1041
    Keywords: Key words Sufentanil ; pharmacokinetics ; haemo dynamics ; different infusion rates ; coronary surgery
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract    Objective: Pharmacokinetics and haemodynamic effects of a total dose of 15 μg · kg−1 sufentanil, an opioid anaesthetic agent, were studied in patients undergoing aortocoronary bypass surgery at three infusion rates of 30 (group I), 5 (group II), and 2 (group III) μg · kg−1 · min−1, respectively. Results: Plasma concentrations of sufentanil could be optimally characterized by a linear biexponential pharmacokinetic model. Non-compartmental analyses indicated that there was no significant difference in the values of clearance (11.6, 13.3, 14.3 ml · min−1 · kg−1), steady-state volume of distribution (0.220, 0.255 and 0.331 l · kg−1) and mean residence time (18.8, 13.3 and 14.3 min) among the groups. The observed mean Cmax values of 421 (group I), 125 (group II), and 53 (group III) ng · ml−1 and observed mean AUC values from 0 to 3 min were all consistent with the dosing regimens. There were large inter-individual variations in haemodynamic response. Compared to plasma data, a delay in haemodynamic effects was found. Times to reach peak haemodynamic effect ranged from 4.3 to 4.9 min for group I, from 4.6 to 6.1 min for group II, and from 9.9 to 11.3 for group III. Except heart rate, peak haemodynamic effects in these study patients generally ranged from 20.9% to 35.2%. Significant reductions in the area under the effect-time profiles of mean arterial blood pressure and systemic vascular resistance were observed in group II and group III, but not in group I. Significant reductions in the area under the effect-time profiles of left ventricular stroke work index were observed in group III only. No effect on heart rate was found in any group. Conclusion: Our findings suggested that a slower infusion rate of sufentanil at a dose of 15 μg · kg−1 tends to give a greater reduction in mean arterial blood pressure, systemic vascular resistance, and left ventricular stroke work index than does a faster infusion rate.
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    URL: http://dx.doi.org/10.1007/s002280050214
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    Effect of route of administration of fluconazole on the interaction between fluconazole and midazolam (1997)
    Springer
    European journal of clinical pharmacology 51 (1997), S. 415-419 
    Details
    ISSN: 1432-1041
    Keywords: Key words Midazolam ; Fluconazole ; CYP3A4 ; interaction ; pharmacokinetics ; pharmacodynamics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: Midazolam is a short-acting benzodiazepine hypnotic extensively metabolized by CYP3A4 enzyme. Orally ingested azole antimycotics, including fluconazole, interfere with the metabolism of oral midazolam during its absorption and elimination phases. We compared the effect of oral and intravenous fluconazole on the pharmacokinetics and pharmacodynamics of orally ingested midazolam. Methods: A double-dummy, randomized, cross-over study in three phases was performed in 9 healthy volunteers. The subjects were given orally fluconazole 400 mg and intravenously saline within 60 min; orally placebo and intravenously fluconazole 400 mg; and orally placebo and intravenously saline. An oral dose of 7.5 mg midazolam was ingested 60 min after oral intake of fluconazole/placebo, i.e. at the end of the corresponding infusion. Plasma concentrations of midazolam, α-hydroxymidazolam and fluconazole were determined and pharmacodynamic effects were measured up to 17 h. Results: Both oral and intravenous fluconazole significantly increased the area under the midazolam plasma concentration-time curve (AUC0–3, AUC0–17) 2- to 3-fold, the elimination half-life of midazolam 2.5-fold and its peak concentration (Cmax) 2- to 2.5-fold compared with placebo. The AUC0–3 and the Cmax of midazolam were significantly higher after oral than after intravenous administration of fluconazole. Both oral and intravenous fluconazole increased the pharmacodynamic effects of midazolam but no differences were detected between the fluconazole phases. Conclusion: We conclude that the metabolism of orally␣administered midazolam was more strongly inhibited by oral than by intravenous administration of fluconazole.
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    URL: http://dx.doi.org/10.1007/s002280050223
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    Lack of interaction between meloxicam and warfarin in healthy volunteers (1997)
    Springer
    European journal of clinical pharmacology 51 (1997), S. 421-425 
    Details
    ISSN: 1432-1041
    Keywords: Key words Warfarin ; Meloxicam ; interaction ; pharmacokinetics ; protein binding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: The effect of multiple oral doses of meloxicam 15 mg on the pharmacodynamics and pharmacokinetics of warfarin was investigated in healthy male volunteers. Warfarin was administered in an individualized dose to achieve a stable reduction in prothrombin times calculated as International Normalized Ratio (INR) values. Then INR- and a drug concentration-time profile was determined. For the interaction phase, meloxicam was added for 7 days and then INR measurements and the warfarin drug profiles were repeated for comparison. Overall, warfarin treatment lasted for 30 days. Results: Warfarin and meloxicam were well tolerated by healthy volunteers in this study. Thirteen healthy volunteers with stable INR values entered the interaction phase. Prothrombin times, expressed as mean INR values, were not significantly altered by concomitant meloxicam treatment, being 1.20 for warfarin alone and 1.27 for warfarin with meloxicam cotreatment. R- and S-warfarin pharmacokinetics were similar for both treatments. Geometric mean (% gCV) AUCSS values for the more potent S-enantiomer were 5.07 mg · h · l−1 (27.5%) for warfarin alone and 5.64 mg · h · l−1 (28.1%) during the interaction phase. Respective AUCSS values for R-warfarin were 7.31 mg · h · l−1 (43.8%) and 7.58 mg · h · l−1 (39.1%). Conclusion: The concomitant administration of the new non-steroidal anti-inflammatory drug (NSAID) meloxicam affected neither the pharmacodynamics nor the pharmacokinetics of a titrated warfarin dose. A combination of both drugs should nevertheless be avoided and, if necessary, INR monitoring is considered mandatory.
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    URL: http://dx.doi.org/10.1007/s002280050224
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    Genetic polymorphism of CYP2C19 and lansoprazole pharmacokinetics in Japanese subjects (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 391-396 
    Details
    ISSN: 1432-1041
    Keywords: Key words Lansoprazole ; CYP2C19; genotype ; hydroxy lation ; polymorphism ; pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: We investigated whether interindividual differences in the pharmacokinetic disposition of lansoprazole are attributed to the genetic polymorphism of CYP2C19 which occurred by two mutations, CYP2C19m1 and CYP2C19m2, in 20 Japanese subjects. Methods: Polymerase chain reaction (PCR) restriction fragment length polymorphism procedures were used to detect the CYP2C19m1 mutation in exon 5 and the CYP2C19m2 mutation in exon 4 using SmaI and BamHI, respectively. Results: Ten subjects were homozygous (wt/wt subjects) for the wt allele in both exon 5 and exon 4, four subjects were heterozygous (wt/m1) for the CYP2C19m1 mutation, and two subjects were heterozygous (wt/m2) for the CYP2C19m2. The remaining four subjects had both mutated alleles in CYP2C19 genes, i.e., two were homozygous (m1/m1) for the defect in exon 5 and two were heterozygous (m1/m2) for the two defects in exons 5 and 4. The subjects in group 1 (wt/wt, wt/m1 and wt/m2) were the extensive metabolizers (EMs) for 5-hydroxylation of lansoprazole and were in the range of hydroxylation indexes from 3.83 to 19.8, whereas the subjects in group 2 (m1/m1 and m1/m2) were the poor metabolizers (PMs) and the indexes were in the range of 38.5 to 47.6. In group 2, AUC, t1/2 and CL/f of lansoprazole were significantly greater, longer, and lower, respectively, than those in group 1. Conclusion: The hydroxylation of lansoprazole to 5-hydroxylansoprazole was apparently impaired in the subjects with the genetic defects of CYP2C19 (m1/m1 or m1/m2).
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    URL: http://dx.doi.org/10.1007/s002280050307
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    Effects of grapefruit juice ingestion – pharmacokinetics and haemodynamics of intravenously and orally administered felodipine in healthy men (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 139-145 
    Details
    ISSN: 1432-1041
    Keywords: Key words Felodipine ; Dietary interaction ; Flavonoids; pharmacodynamics ; pharmacokinetics ; grapefruit juice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: To examine the effect of grapefruit juice on the metabolism of felodipine following intravenous and oral administration. Methods: The study had a randomised, four-way, crossover design in 12 healthy males. Single doses of felodipine were given as an intravenous infusion for 1 h (1.5 mg) or as an oral extended release (ER) tablet (10 mg). Grapefruit juice (150 ml) or water was ingested 15 min prior to drug intake. Results: Intake of grapefruit juice did not significantly alter the intravenous pharmacokinetics of felodipine compared to control treatment, whereas after oral drug administration it did lead to an increase in the mean AUC and Cmax by 72% and 173%, respectively, and the mean absolute bioavailability was increased by 112%. The fraction of the oral felodipine dose reaching the portal system was increased from 45% to 80% when intake of drug was preceded by grapefruit juice ingestion. The pharmacokinetics of the primary metabolite, dehydrofelodipine, was affected by the intake of juice, resulting in a 46% increase in Cmax. Juice intake immediately before oral felodipine resulted in more pronounced haemodynamic effects of the drug as measured by diastolic blood pressure and heart rate. However, the haemodynamic effects of the intravenous administration were not altered by juice intake. Vascular-related adverse events were reported more frequently when oral drug administration was preceded by juice intake compared with control treatment. Taking grapefruit juice immediately prior to intravenous felodipine administration did not cause any alteration in the adverse event pattern. Conclusion: The main acute effect of the grapefruit juice on the plasma concentrations of felodipine is mediated by inhibition of gut wall metabolism.
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    URL: http://dx.doi.org/10.1007/s002280050263
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    Pharmacokinetics and bioavailability of oral and intramuscular artemether (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 307-310 
    Details
    ISSN: 1432-1041
    Keywords: Key words Artemether ; Thai males; malaria ; dihydroartemisinin ; pharmacokinetics ; bioavailability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: The pharmacokinetics and bioavailability of artemether and dihydroartemisinin were investigated in eight Thai males following the administration of single oral and intramuscular doses of artemether (300 mg) in a randomized two-way cross-over study. Results: Both oral and intramuscular artemether were well-tolerated. In most cases, artemether and dihydroartemisinin were detected in plasma after 30 min and declined to levels below the limit of detection within 18–24 h. Compared with intramuscular administration, oral administration of artemether resulted in a relatively rapid but incomplete absorption [Cmax: 474 vs 540 ng · ml−1; t max: 2.0 vs 3.9 h; AUC: 2.17 vs 5.20 μg · h · ml−1]. Geographic means of lag-time and absorption half-life (t 1/2a) of oral vs intramuscular artemether were 0.28 and 1.1 h vs 0.30 and 2 h, respectively. t 1/2z was significantly shortened after the oral dose [2.8 vs 6.9 h]. Mean oral bioavailability relative to intramuscular administration was 43.2%. The ratio of the AUCs of artemether to dihydroartemisinin was significantly lower after the oral than after the intramuscular dose (geometric mean: 0.29 vs 0.60).
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    URL: http://dx.doi.org/10.1007/s002280050295
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    Itraconazole moderately increases serum concentrations of oxybutynin but does not affect those of the active metabolite (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 403-406 
    Details
    ISSN: 1432-1041
    Keywords: Key words Oxybutynin ; Itraconazole; N-desethyloxy‐butynin; drug interaction ; pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: Oxybutynin has low oral bioavailability due to an extensive presystemic metabolism. It has been suggested that the biotransformation of oxybutynin is dependent on CYP3A. Because itraconazole, a widely used mycotic, is a potent inhibitor of CYP3A4, we wanted to study a possible interaction between oxybutynin and itraconazole. Methods: In this double-blind, randomized, two-phase cross-over study, ten healthy volunteers received either 200 mg itraconazole or placebo for 4 days. On day 4, each volunteer ingested a single dose of 5 mg oxybutynin. Serum concentrations of oxybutynin, its active metabolite N-desethyloxybutynin, and itraconazole were monitored over 24 h. Results: Itraconazole significantly increased both the area under the serum drug concentration-time curve (AUC0–t) and the peak concentration of oxybutynin twofold. The AUC0–t and the peak concentration of N-desethyloxybutynin were not significantly affected by itraconazole. Itraconazole did not change the peak time or the elimination half-life of either oxybutynin or N-desethyloxybutynin. The occurrence of adverse events after oxybutynin administration was not increased by itraconazole. Conclusions: Itraconazole moderately increases serum concentrations of oxybutynin, probably by inhibiting the CYP3A-mediated metabolism. However, the concentrations of N-desethyloxybutynin were practically unchanged. Since about 90% of the antimuscarinic activity of oxybutynin is attributable to N-desethyloxybutynin, any interaction of oxybutynin with CYP3A4 inhibiting drugs has only minor clinical significance.
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    URL: http://dx.doi.org/10.1007/s002280050309
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    Population analysis of the non-linear red blood cell partitioning of draflazine following various infusion durations (1997)
    Springer
    European journal of clinical pharmacology 53 (1997), S. 57-63 
    Details
    ISSN: 1432-1041
    Keywords: Key wordsDraflazine ; Population analysis; nucleoside transport inhibitor ; non-linear red blood cell partition ing ; pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: The pharmacokinetics and non-linear red blood cell partitioning of the nucleoside transport inhibitor draflazine were investigated in 19 healthy male and female subjects (age range 22–55 years) after a 15-min i.v. infusion of 1 mg, immediately followed by infusions of variable rates (0.25, 0.5 and 1 mg · h−1) and variable duration (2–24 h). Methods: The parameters describing the capacity-limited specific binding of draflazine to the nucleoside transporters located on erythrocytes were determined by NONMEM analysis. The red blood cell nucleoside transporter occupancy of draflazine (RBC occupancy) was evaluated as a pharmacodynamic endpoint. Results: The population typical value for the dissociation constant K d (%CV) was 0.648 (12) ng · ml−1 plasma, expressing the very high affinity of draflazine for the erythrocytes. The typical value of the specific maximal binding capacity Bmax (%CV) was 155 (2) ng · ml−1 RBC. The interindividual variability (%CV) was moderate for K d (38.9%) and low for Bmax (7.8%). As a consequence, the variability in RBC occupancy of draflazine was relatively low, allowing the justification of only one infusion scheme for all subjects. The specific binding of draflazine to the red blood cells was a source of non-linearity in draflazine pharmacokinetics. Steady-state plasma concentrations of draflazine virtually increased dose-proportionally and steady state was reached at about 18 h after the start of the continuous infusion. The t1/2βaveraged 11.0–30.5 h and the mean CL from the plasma was 327 to 465 ml · min−1. The disposition of draflazine in whole blood was different from that in plasma. The mean t1/2β was 30.2 to 42.2 h and the blood CL averaged 17.4–35.6 ml · min−1. Conclusion: Although the pharmacokinetics of draflazine were non-linear, the data of the present study demonstrate that draflazine might be administered as a continuous infusion over a longer time period (e.g., 24 h). During a 15-min i.v. infusion of 1 mg, followed by an infusion of 1 mg · h−1, the RBC occupancy of draflazine was 96% or more. As the favored RBC occupancy should be almost complete, this dose regimen could be justified in patients.
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    Single-dose pharmacokinetics of paracetamol and its conjugates in Chinese non-insulin-dependent diabetic patients with renal impairment (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 285-288 
    Details
    ISSN: 1432-1041
    Keywords: Key words Paracetamol ; Renal failure; polar conjugates ; non-insulin-dependent diabetes mellitus (NIDDM) ; pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: A single oral dose of paracetamol (20 mg · kg−1) was given to 38 Chinese patients with non-insulin-dependent diabetes mellitus (NIDDM) who had either normal renal function or varying degrees of renal impairment, with creatinine clearances ranging from 4 to 123 ml · min−1 · 1.73 m−2. The plasma and urinary concentrations of paracetamol and its major metabolites were measured by high-performance liquid chromatography (HPLC). Results: The absorption and elimination of paracetamol were unaffected by renal impairment. However, the area under the plasma concentration time curve and the elimination half-life of paracetamol metabolites increased significantly with worsening renal insufficiency. Mean renal clearances of paracetamol and its conjugates were significantly reduced in these subjects. There was no evidence of altered metabolic activation with renal impairment. Conclusion: The results demonstrate that paracetamol disposition is minimally affected by diabetic nephropathy; however, extensive accumulation of conjugates may occur.
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    URL: http://dx.doi.org/10.1007/s002280050291
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    Pharmacokinetics and pharmacodynamics of ranitidine in renal impairment (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 229-234 
    Details
    ISSN: 1432-1041
    Keywords: Key words Ranitidine ; Renal impairment; dose adjustment ; pharmacodynamics ; pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: The pharmacodynamics and pharmacokinetics of ranitidine were examined in subjects with varying degrees of renal function to determine the effect of this condition on acid-antisecretory activity. Methods: Subjects with creatinine clearances (CCr) ranging from 0 to 213 ml · min−1 received single 50-mg and 25-mg i.v. doses of ranitidine. This was followed by determination of serum and urine ranitidine concentrations, and continuous gastric pH monitoring for 24 h. Results: Serum ranitidine concentrations were described by a two-compartment model linked to a sigmoidal Emax model describing gastric pH. Ranitidine renal clearance, ranging from 0 to 1003 ml · min−1, correlated with CPAH (r 2 = 0.707), while non-renal clearance was unaltered. Steady-state volume of distribution decreased by half in severe renal impairment. No changes in the effective concentration at half-maximal response (EC50), maximal response (Emax), or basal response (E0) were observed. Thus, renal elimination of ranitidine declined in parallel with renal function, while sensitivity to the pharmacologic effect (gastric pH elevation) was unaltered. Ranitidine was well tolerated in these renally impaired subjects. Conclusion: These data indicate that the current recommendation for renal impairment dose reduction (by two-thirds when CCr〈50 ml · min−1) might result in under-treating moderately impaired patients, and suggests a less conservative dose reduction (by half when CCr〈10 ml · min−1) to avoid therapeutic failure while remaining within the wide margin of safety for this drug.
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    URL: http://dx.doi.org/10.1007/s002280050279
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    Pharmacokinetic interaction study of citalopram and cimetidine in healthy subjects (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 241-242 
    Details
    ISSN: 1432-1041
    Keywords: Key words Citalopram ; Cimetidine; drug ; drug interac‐tion ; pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002280050282
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    Pharmacokinetic-pharmacodynamic modelling as a tool to evaluate the clinical relevance of a drug-food interaction for a nisoldipine controlled-release dosage form (1997)
    Springer
    European journal of clinical pharmacology 51 (1997), S. 473-480 
    Details
    ISSN: 1432-1041
    Keywords: Key words Nisoldipine ; Hypertension; Ca antagonist ; pharmacokinetics ; pharmacodynamics ; PK/PD modelling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Objective: Nisoldipine, a calcium antagonist of the dihydropyridine class, has been used in the treatment of hypertension and angina pectoris. A new controlled-release dosage form (nisoldipine coat-core, NCC) has been developed to allow once daily dosing. In addition to a formal food interaction study as requested by regulatory authorities for controlled-release dosage forms, a subsequent study was conducted to determine the clinical relevance of the changes in nisoldipine plasma concentration vs time profiles seen in the food effect study. Methods: After a placebo run-in phase of 6 days, 12 hypertensive patients started treatment with 20 mg NCC once daily (days 0–3, 5–6, 8–9). On days 4, 7 and 10 the NCC was substituted for 5, 10 and 20 mg nisoldipine solution, respectively, in order to obtain nisoldipine plasma concentration vs time profiles comparable to the ones resulting from the concomitant intake of food and NCC. Simultaneous measurements of blood pressure (BP) and nisoldipine concentration were performed on days 3, 4, 7 and 10. Results: The relationship between nisoldipine plasma concentrations and percentage reduction in BP [diastolic (DBP) and systolic (SBP), supine and standing] could be described by an Emax model. The mean maximum reduction (Emax) relative to baseline was about 36.4% and 37.7% (DBP, supine and standing) and 27.9% and 29.2% (SBP, supine and standing), respectively. The interindividual variability (% CV) in Emax was low, ranging from 17.6% to 28.8%. The mean nisoldipine plasma concentration corresponding to 50% of the maximum effect (EC50) ranged between 0.99 and 2.62 μg · l–1 with a pronounced interindividual variability (% CV) of 89.5–108.8%. Mean Cmax values after administration of the 30 and 40 mg NCC together with food were 4.5 and 7.5 μg · l–1, respectively. Based on the concentration-effect relationship established in the present study, the effect achieved with a concentration of 7.5 μg · l–1 will be about 77% of Emax for DBP and about 88% of Emax for SBP, respectively. Conclusion: At the time of maximum plasma concentration the additional decrease in BP relative to baseline due to the food effect will be about 7–15% for DBP and 3–9% for SBP. After administration of the 10␣mg solution with a mean Cmax of 8.7 μg · l–1, only headache and flush with mild severity have been reported as adverse events. These maximum concentrations are comparable to Cmax values seen after intake of 40 mg NCC with food. With regard to heart rate (HR) there were distinct differences between the two formulations: Following administration of 5, 10 and 20 mg nisoldipine solution, there were dose-dependent increases in HR by a maximum of 4, 12 and 16 beats · min−1, respectively, whereas the HR profile for the NCC was similar to that seen under placebo treatment.
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    URL: http://dx.doi.org/10.1007/s002280050233
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    Calreticulin inhibits glucocorticoid– but not cAMP–sensitive expression of tyrosine aminotransferase gene in cultured McA–RH7777 hepatocytes (1997)
    Springer
    Molecular and cellular biochemistry 171 (1997), S. 37-43 
    Details
    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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    URL: http://dx.doi.org/10.1023/A:1006865108833
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    Stimulatory effect of calcium administration on regucalcin mRNA expression is attenuated in the kidney cortex of rats ingested with saline (1998)
    Springer
    Molecular and cellular biochemistry 178 (1998), S. 275-281 
    Details
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; calmodulin ; spontaneous hypertensive rats ; rat kidney cortex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The expression of calcium-binding protein regucalcin mRNA in the kidney cortex of rats ingested with saline was investigated. The alteration in regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb of open reading frame). Rats were freely given saline as drinking water for 7 days. Regucalcin mRNA levels in the kidney cortex were suppressed by saline ingestion. When calcium chloride (10 mg Ca/100 g body weight) was intraperitoneally administered to rats ingested with saline for 7 days, the effect of calcium administration to increase regucalcin mRNA levels was weakened by saline ingestion. Such effect was also seen by the administration of 2.5 and 5 mg Ca/100 g. Regucalcin mRNA levels in the kidney cortex of spontaneous hypertensive rats (SHR) were not appreciably increased by the administration of calcium (10 mg/100 g). Meanwhile, calcium content in the kidney cortex was significantly elevated by the administration of calcium (10 mg/100 g) to normal rats. This increase was weakened in saline-ingested rats. Moreover, Ca2+/calmodulin-dependent protein kinase activity in the cytosol of kidney cortex was significantly decreased by saline ingestion. These results suggest the possibility that saline ingestion-induced suppression of regucalcin mRNA expression in the kidney cortex is partly involved in the attenuation of Ca2+ signalling.
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    Hepatic calcium-binding protein regucalcin concentration is decreased by streptozotocin-diabetic state and ethanol ingestion in rats (1997)
    Springer
    Molecular and cellular biochemistry 168 (1997), S. 67-72 
    Details
    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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    Tamoxifen enhances interferon-regulated gene expression in breast cancer cells (1997)
    Springer
    Molecular and cellular biochemistry 167 (1997), S. 169-177 
    Details
    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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    Use of a hammerhead ribozyme with cationic liposomes to reduce leukocyte type 12-lipoxygenase expression in vascular smooth muscle (1997)
    Springer
    Molecular and cellular biochemistry 172 (1997), S. 47-57 
    Details
    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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    Construction of a human heart cDNA library and identification of cardiovascular based genes (CVBest) (1997)
    Springer
    Molecular and cellular biochemistry 172 (1997), S. 81-87 
    Details
    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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    Hydrogen peroxide-induced expression of the proto-oncogenes, c-jun, c-fos and c-myc in rabbit lens epithelial cells (1997)
    Springer
    Molecular and cellular biochemistry 173 (1997), S. 59-69 
    Details
    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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    Molecular cloning of the cDNA coding for regucalcin and its mRNA expression in mouse liver: The expression is stimulated by calcium administration (1997)
    Springer
    Molecular and cellular biochemistry 173 (1997), S. 127-133 
    Details
    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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    Cardiac hypertrophy: Old concepts, new perspectives (1997)
    Springer
    Molecular and cellular biochemistry 176 (1997), S. 273-279 
    Details
    ISSN: 1573-4919
    Keywords: cardiac hypertrophy ; myosin heavy chain ; gene expression ; adrenergic system
    Source: Springer Online Journal Archives 1860-2000
    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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    URL: http://dx.doi.org/10.1023/A:1006865515646
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    The Glycophorin A Gene Family in Gorillas: Structure, Expression, and Comparison with the Human and Chimpanzee Homologues (1997)
    Springer
    Biochemical genetics 35 (1997), S. 59-76 
    Details
    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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    URL: http://dx.doi.org/10.1023/A:1022212630370
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    Novel methods for adenovirus-mediated gene transfer to blood vessels in vivo (1997)
    Springer
    Molecular and cellular biochemistry 172 (1997), S. 37-46 
    Details
    ISSN: 1573-4919
    Keywords: gene transfer ; gene expression ; adenovirus ; blood vessel
    Source: Springer Online Journal Archives 1860-2000
    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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    Differential display: Identifying genes involved in cardiomyocyte proliferation (1997)
    Springer
    Molecular and cellular biochemistry 172 (1997), S. 111-120 
    Details
    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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    URL: http://dx.doi.org/10.1023/A:1006819705814
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    Increased gene expression of plasminogen activators and inhibitors in left ventricular hypertrophy (1997)
    Springer
    Molecular and cellular biochemistry 176 (1997), S. 265-271 
    Details
    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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    Protein tyrosine phosphatase gene expression analysis in Swiss 3T3 fibroblasts (1998)
    Springer
    Molecular and cellular biochemistry 178 (1998), S. 157-162 
    Details
    ISSN: 1573-4919
    Keywords: protein tyrosine phosphatases ; gene expression ; degenerate deoxyoligonucleotides ; RT-PCR ; Swiss 3T3 fibroblasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The aim of this study was to identify protein tyrosine phosphatases (PTPs) expressed in Swiss 3T3 fibroblasts and to examine their expression levels as well as to characterize quantitative aspects of RT-PCR based on degenerate deoxyoligonucleotides. By using an RT-PCR assay based on degenerate deoxyoligonucleotide primers, expression of mRNAs for two cytoplasmic- and six transmembrane-type PTPs in Swiss 3T3 cells was detected. The sequences of two of them are new. Among nine analyzed PTPs expressed to widely varied extends, only three have mRNA levels high enough to be seen on Northern blots with 10 µg of total RNA per lane. The frequencies with which the examined PTPs are represented among the PCR amplification products, correlate stronger with the primer fidelity, defined as the number of mismatches between the primer- and the cDNA target-sequences, rather than with the PTP expression levels. In conclusion, an RT-PCR assay based on degenerate primers can be successfully used to sample the expressed PTPs and to identify new members of this gene family. However, reliable quantification of their mRNA levels can only be achieved using the classical approaches, like Northern, RNase protection assay or non-degenerate quantitative RT-PCR.
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    URL: http://dx.doi.org/10.1023/A:1006897629337
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    Gene expression after short periods of coronary occlusion (1998)
    Springer
    Molecular and cellular biochemistry 186 (1998), S. 43-51 
    Details
    ISSN: 1573-4919
    Keywords: myocardial ischemia ; gene expression ; growth factors ; phospholamban ; calsequestrin heat shock proteins ; preconditioning ; stunning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Brief periods of coronary occlusion render the affected myocardium more tolarant to the otherwise devastating effects of long coronary occlusion. Besides this phenomena, called ischemic preconditioning, short periods of ischemia cause a regional dysfunction, namely myocardial stunning. The molecular mechanisms of both syndromes are not very well understood. We therefore investigated the expression of genes which may be involved in cardioprotection or repair processes.Using our porcine model of ischemia and reperfusion we were able to show an induction of genes coding for transcription factors (proto-oncogenes), for proteins involved in repair processes (heat shock genes), for proteins implicated in the calcium homeostasis (calcium-handling genes) and for growth factors. We could show that the increased mRNA levels are due to an enhanced transcriptional activity and not to a prolonged half-life of the transcripts. The angiogenic growth factor vascular endothelial growth factor (VEGF) represents an exception. It exhibits - in addition to a HIF-motif (Hypoxia Inducible Factor) in its promoter/enhancer - a protein binding region in its 3′ UTR which when occupied renders the mRNA more stable. However to what extent the expression of the distinct genes contributes to the cardioprotective effect of ischemic preconditioning or myocardial stunning can only be presumed. Increased mRNA stability can be confered via adenosine which is produced during ischemia by ATP-breakdown. The demasking of unknown genes - via differential display reverse transcription polymerase chain reaction (DDRT-PCR) - should provide a more comprehensive view of the mechanisms underlying both processes.
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    Characteristics of Hydroxamic Acid Induction in Wheat Triggered by Aphid Infestation (1997)
    Springer
    Journal of chemical ecology 23 (1997), S. 2695-2705 
    Details
    ISSN: 1573-1561
    Keywords: Defense ; herbivory ; aphids ; wheat ; Gramineae ; hydroxamic acids ; Defense theory ; Carbon/Nutrient theory
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Hydroxamic acids (Hx) are natural products of Gramineae that are associated with cereal resistance to pests. We aimed at characterizing the induction of Hx accumulation in seedlings of wheat,Triticum aestivum, by short-term infestation of the cereal aphid,Rhopalosiphum padi. A load of 25 aphids increased significantly the Hx levels in the infested primary leaf in comparison with control levels. Lower loads did not increase Hx concentration. Aphid infestation lasting 16 hr did not elicit induction of Hx, even after a time-lag of 32 hr to allow the expression of any induced response. Forty-eight hours was the minimum duration of aphid infestation required to trigger Hx induction. The age of the infested tissue (the primary leaf) did not affect induction. Similar increases of Hx were found in unfolding, expanding, and totally expanded primary leaves. It was determined that the regime of nutrient supply (N-intensive nutritive solutions at low and high concentration) to wheat seedlings had no effect on the magnitude of the aphid-induced Hx (N-based secondary metabolites). Results obtained are discussed in the framework of general theories of plant defense allocation.
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    Influence of Taxonomy, Ecology, and Seasonality in River Stage on Fish Contamination Risks in Floodplain Swamps of the Lower Mississippi River (1998)
    Springer
    Ecotoxicology 7 (1998), S. 325-334 
    Details
    ISSN: 1573-3017
    Keywords: contamination risks ; fish ; Mississippi River ; ecological factors ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract We compared contamination levels in fish from contaminated and uncontaminated floodplain swamps of the lower Mississippi River to assess differences in contamination risks between swamps, across different taxonomic and ecological groupings of fishes within and between swamps, and with seasonality in river stage. Fish tissue levels of inorganic contaminants were substantially lower than environmental levels in both swamps, suggesting either that fish were not uptaking these contaminants, or they were effectively eliminating the contaminants from their bodies. Tissue levels of organic contaminants were high relative to environmental levels, suggesting that these contaminants were bioaccumulating. Organic contaminants were significantly higher in fish from the contaminated swamp (Devil's Swamp) than in fish from a reference swamp up river (Tunica Swamp). Because the organic contaminants were largely confined to sediments, we expected bottom-oriented fishes to have higher concentrations than pelagic fishes. Assuming that uptake was primarily through the food chain, we expected top predators to exhibit higher concentrations than low-level consumers. We also expected year- round swamp residents to exhibit higher accumulations than more transitory users of backswamp habitat. However, organic contaminant levels did not differ in the directions expected for any of these groupings. We did observe differences in organic contaminant levels within and between swamps for different taxonomic groupings of fishes (species and genera). Some taxa occupying low to middle positions in the food web (e.g., gizzard shad, Lepomis spp.) exhibited higher concentrations than taxa near the top of the food web. Within Devil's Swamp, organic contaminant levels were significantly higher at low river stage, when fish were confined to the swamp, than at high river stage, when fish were free to move between the river and the swamp. We caught more species and more fish per unit effort in Devil's Swamp than in Tunica Swamp, contrary to expectations if contaminants in the former were negatively impacting population and community structure. Species richness differences between swamps were a consequence of catch differences, with higher catch corresponding to inclusion of more rare species. The lower catch in Tunica Swamp may have resulted from physical modifications of its waterways to support agriculture and hunting. The results of this study underscore the importance in factoring information on the taxonomy and ecology of organisms, and seasonal changes in environmental conditions, into assessments of contamination risks.
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    URL: http://dx.doi.org/10.1023/A:1008811713427
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    Regulation of lipoprotein metabolism by estrogen in inbred strains of mice occurs primarily by posttranscriptional mechanisms (1997)
    Springer
    Molecular and cellular biochemistry 173 (1997), S. 161-168 
    Details
    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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    Expression of calcium-binding protein regucalcin mRNA in the cloned human hepatoma cells (HegG2): Stimulation by insulin (1997)
    Springer
    Molecular and cellular biochemistry 175 (1997), S. 163-168 
    Details
    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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    Intrinsic secondary structure of human TNFR-I mRNA influences the determination of gene expression by RT-PCR (1997)
    Springer
    Molecular and cellular biochemistry 177 (1997), S. 1-6 
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    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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    URL: http://dx.doi.org/10.1023/A:1006862304381
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    Alternative splicing of human plasma cholesteryl ester transfer protein mRNA in Caco-2 cells and its modulation by oleic acid (1997)
    Springer
    Molecular and cellular biochemistry 177 (1997), S. 107-112 
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    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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    Expression of calcium-binding protein regucalcin mRNA in fetal rat liver is stimulated by calcium administration (1998)
    Springer
    Molecular and cellular biochemistry 178 (1998), S. 283-287 
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    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; fetal development ; rat liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The expression of hepatic calcium-binding protein regucalcin mRNA in fetal rats was investigated. The alteration in regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin cDNA (0.9 kb with complete open reading frame). Hepatic regucalcin mRNA levels were progressively increased with fetal development; the mRNA was clearly expressed at 15 and 21 days of pregnancy but only slightly at the 8 days. Meanwhile, β-actin mRNA levels in the fetal liver were remarkable at 8 and 15 days of pregnancy. The fetal liver regucalcin mRNA levels at 15 days of pregnancy were significantly decreased by overnight-fasting of maternal rats. The oral administration of calcium chloride (50 mg Ca/100 g body weight) to maternal rats at 15 days of pregnancy caused a remarkable elevation (about 2 fold) of regucalcin mRNA levels in the fetal liver; this increase was seen 60 and 180 min after the calcium administration. After birth, regucalcin mRNA was increasingly expressed in the livers of newborn and weanling rats, while hepatic β-actin mRNA expression was not appreciably altered with increasing ages. These findings demonstrate that the expression of hepatic regucalcin mRNA is increased with fetal development, and that the gene expression may be stimulated by the ingestion of dietary calcium.
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    Enhanced NOR-1 gene expression by exposure of Chinese hamster cells to high-density 50 Hz magnetic fields (1998)
    Springer
    Molecular and cellular biochemistry 181 (1998), S. 191-195 
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    ISSN: 1573-4919
    Keywords: extremely low frequency magnetic fields ; gene expression ; neuron derived orphan receptor-1 ; signal transduction ; Chinese hamster ovary K1 cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Enhanced expression of neuron derived orphan receptor (NOR-1) gene was observed by exposure of Chinese hamster ovary K1 (CHO-K1) cells to an extremely low frequency magnetic field (ELFMF) of 50 Hz at 400 mT, but not at 5 mT. The enhanced expression, reaching the maximum at 6 h, was transient and reduced to the control level after exposure to 400 mT ELFMF for 24 h. The NOR-1 expression induced by treatment with forskolin and TPA was further enhanced by the simultaneous treatment with 400 mT ELFMF, in which the maximum response was at 3 h. The NOR-1 expression by these treatments was induced more earlier than that by 400 mT ELFMF alone. When cells were treated with an inhibitor of the protein kinase C (calphostin C or crocetin) and Ca2+ entry blockers (nifedipin and dantrolen) during the 400 mT ELFMF exposure, the enhanced NOR-1 expression was not observed. Exposure of CHO-K1 cells to the high-density 400 mT ELFMF may affect the signal transduction in the cells, resulting in the enhanced NOR-1 gene expression.
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    The molecular basis for the role of zinc in developmental biology (1998)
    Springer
    Molecular and cellular biochemistry 188 (1998), S. 41-48 
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    ISSN: 1573-4919
    Keywords: zinc ; transcription factors ; gene expression ; organogenesis ; Xenopus laevis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Zinc regulates the gene expression machinery. It affects the structure of chromatin, the template function of its DNA, the activity of numerous transcription factors and of RNA polymerases. Hence, it determines both the types of mRNA transcripts synthesized and the rate of transcription itself. Alterations in one or more of these zinc dependent processes have been proposed to account for the proliferative arrest and teratology induced by zinc deficiency. To examine this proposal, studies of zinc during X. laevis development have been initiated. The kinetics of X. laevis oocyte zinc uptake and storage and of zinc utilization during embryogenesis have been examined first. Vitellogenin carries zinc into the oocyte. Ten % of the total zinc (10 ng/egg) remains within the cytosol while 90% (90 ng/egg) is stored in the yolk platelets associated with lipovitellin. The cytosolic pool is the source of the zinc for all newly formed metalloproteins involved in embryo development. The yolk platelet zinc pool is stored for later use during early metamorphosis. It is now possible to examine zinc transfer to molecules, such as e.g. transcription factors, and the role of the metal in their function in development and organogenesis.
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    Effect of 60 Hz magnetic field exposure on c-fos expression in stimulated PC12 cells (1998)
    Springer
    Molecular and cellular biochemistry 189 (1998), S. 107-111 
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    ISSN: 1573-4919
    Keywords: gene expression ; electromagnetic fields ; superinduction ; anisomycin ; immediate early gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Rat pheochromocytoma PC12 cells have been treated with nerve growth factor (NGF) at final concentrations of 2, 4, 8, and 16 ng/ml, and then were exposed to 60-Hz, sinusoidal magnetic fields (MF) of 12.5, 25, 50, and 100 μT (rms) for 30 min. Transcript levels for both c-fos and glyceraldehyde-3 -phosphate dehydrogenase were determined by Northern blot analysis using 32P-labeled cDNA probes. No change in c-fos expression was measured at any condition employed. Treatment of PC12 cells with a combination of agents (NGF, forskolin, and tetradecanoylphorbol acetate [TPA]) increased c-fos expression over that detected with NGF alone. MF exposure of cells treated with the three-agent regimen produced two outcomes, either no change or a doubling of c-fos expression. In subsequent experiments, cells were treated with NGF, NGF + forskolin + TPA, or pre-treated with anisomycin and then treated with NGF + forskolin + TPA. It was determined that MF exposure, like superinduction with anisomycin, increased c-fos expression only in cultures which were not yet exhibiting maximal c-fos expression. It is hypothesized that MF exposure, like anisomycin, may alter the activity of key intracellular protein kinases.
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    Expression of mouse RNase MRP RNA in human embryonic kidney 293 cells (1997)
    Springer
    Molecular biology reports 24 (1997), S. 221-230 
    Details
    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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    Nuclear matrix associated DNA-binding proteins of ocular lens epithelial cells (1998)
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    Molecular biology reports 25 (1998), S. 13-19 
    Details
    ISSN: 1573-4978
    Keywords: gene expression ; nuclear matrix proteins ; ocular lens epithelial cells ; transcription factors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Association of transcription factors with the nuclear matrix represents a mechanism by which nuclear architecture may influence transcriptional control of gene expression. This investigation examines nuclear matrix associated proteins (NMP's) isolated from ocular lens epithelial cells by monitoring DNA binding activities using consensus oligonucleotides recognized by the transcription factors YY1, AML-1, AP-1, SP-1 and ATF. The nuclear matrix fractions tested included an immortilized human lens epithelial cell line containing the SV40 large T-antigen, and two mouse lens epithelial cell lines derived from either a normal mouse or a cataract mouse. A rabbit epidermal epithelial cell line and HeLa cells were also included in this study for comparison. The data from these experiments reveal that ubiquitously represented and tissue restricted regulatory proteins are associated with nuclear matrix of lens epithelial cells. The functional significance of the nuclear matrix association of these transcription factors remains to be determined. However, our findings raise the possibility that the transcription factors associated with the nuclear matrix could have specific roles in gene regulation and eye tissue development.
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    Characterization of two cDNA clones for mRNAs expressed during ripening of melon (Cucumis melo L.) fruits (1997)
    Springer
    Plant molecular biology 33 (1997), S. 313-322 
    Details
    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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    Differential transcript induction of parsley pathogenesis-related proteins and of a small heat shock protein by ozone and heat shock (1997)
    Springer
    Plant molecular biology 33 (1997), S. 343-350 
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    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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    Molecular cloning and expression of two cDNAs encoding asparagine synthetase in soybean (1997)
    Springer
    Plant molecular biology 33 (1997), S. 301-311 
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    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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    Regulation of two loblolly pine (Pinus taeda L.) isocitrate lyase genes in megagametophytes of mature and stratified seeds and during postgerminative growth (1997)
    Springer
    Plant molecular biology 33 (1997), S. 593-604 
    Details
    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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    Characterisation of a pea hsp70 gene which is both developmentally and stress-regulated (1997)
    Springer
    Plant molecular biology 34 (1997), S. 345-352 
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    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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    Leaf senescence in Brassica napus: cloning of senescence related genes by subtractive hybridisation (1997)
    Springer
    Plant molecular biology 33 (1997), S. 821-834 
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    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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    Self-incompatibility in the grasses: evolutionary relationship of the S gene from Phalaris coerulescens to homologous sequences in other grasses (1997)
    Springer
    Plant molecular biology 34 (1997), S. 223-232 
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    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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    A chloroplast derived trnH gene is expressed in the mitochondrial genome of gramineous plants (1997)
    Springer
    Plant molecular biology 34 (1997), S. 353-356 
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    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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    Expression of a wheat ADP-glucose pyrophosphorylase gene during development of normal and water-stress-affected anthers (1997)
    Springer
    Plant molecular biology 34 (1997), S. 445-453 
    Details
    ISSN: 1573-5028
    Keywords: ADP-glucose pyrophosphorylase ; anther ; pollen ; male sterility ; water stress ; wheat ; starch
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In wheat (Triticum aestivum L.), water deficit during meiosis in the microspore mother cells (MMCs) induces pollen abortion, resulting in the failure of fertilization and a reduction in grain set. In stressed plants, meiosis in MMCs proceeds normally but subsequent pollen development is arrested. Unlike normal pollen grains, which accumulate starch during the late maturation phase, stress-affected anthers contain pollen grains with little or no starch. Stress also alters the normal distribution of starch in the anther wall and connective tissue. To determine how starch biosynthesis is regulated within the developing anthers of stressed plants, we studied the expression of ADP-glucose pyrophosphorylase (AGP), which catalyzes the rate limiting step of starch biosynthesis. Two partial-length cDNAs corresponding to the large subunit of AGP were amplified by RT-PCR from anther RNA, and used as probes to monitor AGP expression in developing anthers of normal and water-stressed plants. These clones, WAL1 and WAL2, had identical deduced amino acid sequences and shared 96% sequence identity at the nucleic acid level. In normal anthers, AGP expression was biphasic, indicating that AGP expression is required for starch biosynthesis both during meiosis and later during pollen maturation. AGP expression in stressed anthers was not affected during the first phase of starch accumulation, but was strongly inhibited during the second phase. We conclude from these results that the reduced starch deposition later in the development of stressed pollen could be the result of a lower expression of AGP. However, this inhibition of AGP expression is unlikely to be the primary cause of male sterility because anatomical symptoms of pollen abortion are observed prior to the time when AGP expression is inhibited.
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    URL: http://dx.doi.org/10.1023/A:1005882118506
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    Specific sequence modifications of a cry3B endotoxin gene result in high levels of expression and insect resistance (1997)
    Springer
    Plant molecular biology 34 (1997), S. 485-496 
    Details
    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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    LhcaR1 of the red alga Porphyridium cruentum encodes a polypeptide of the LHCI complex with seven potential chlorophyll a-binding residues that are conserved in most LHCs (1997)
    Springer
    Plant molecular biology 33 (1997), S. 157-167 
    Details
    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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    Pollen embryogenesis (1997)
    Springer
    Plant molecular biology 33 (1997), S. 1-10 
    Details
    ISSN: 1573-5028
    Keywords: androgenesis ; anther and microspore culture ; gene expression ; haploids ; pollen embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1005748614261
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    Characterization of dynamics of the psbD light-induced transcription in mature wheat chloroplasts (1997)
    Springer
    Plant molecular biology 33 (1997), S. 267-278 
    Details
    ISSN: 1573-5028
    Keywords: chloroplast ; in vitro transcription ; light ; psbA ; psbD/C operon ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dynamical aspects of three chloroplast promoters responding to change in light condition were examined in mature chloroplasts of wheat (Triticum aestivum) by in vitro transcription. The wheat psbD/C operon has four distinct promoters, two of which named as D/C-3 and D/C-4 promoters dominantly function in mature chloroplasts to produce the mRNAs encoding D2/CP43 and CP43 alone, respectively. Activity of the D/C-3 promoter in mature chloroplasts was reduced to less than 30% by 24 h dark adaptation and recovered by re-illumination to the original level within 30 to 60 min. The activation of the D/C-3 promoter which requires de novo cytoplasmic protein synthesis was induced by low fluence of light (e.g. 16 µE m-2 s-1), but the extent of activation increased with increasing light fluence. The accumulation of mRNAs from the D/C-3 promoter saturated at 2- to 3-fold higher level within 2 h when the dark-adapted seedlings were transferred to the lig at 72 µE m-2 s-1, concomitant with the increase in rate of D2 synthesis, suggesting that synthesis of D2 in mature chloroplasts is controlled via the D/C-3 promoter activity in a light-dependent way. Activity of the D/C-4 promoter slightly increased in the dark and decreased in the light. Effect of light on the psbA promoter activity was not observed at all in mature chloroplasts. In vitro transcriptional analysis of the D/C-3 promoter with 5′ deletion mutations revealed that at least two cis elements which are located within the sequences of -78 to -47 and -46 to -29 of the transcription initiation site, respectively, act as enhancing elements in the D/C-3 promoter. The light-switching element of the transcription, however, was suggested to be located in the core promoter sequence downstream of the -35 element.
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    Effects of chilling on the expression of ethylene biosynthetic genes in Passe-Crassane pear (Pyrus communis L.) fruits (1997)
    Springer
    Plant molecular biology 33 (1997), S. 847-855 
    Details
    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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    Characterization of the gene for Δ1-pyrroline-5-carboxylate synthetase and correlation between the expression of the gene and salt tolerance in Oryza sativa L. (1997)
    Springer
    Plant molecular biology 33 (1997), S. 857-865 
    Details
    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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    Calcium influx signals normal flagellar RNA induction following acid shock of Chlamydomonas reinhardtii (1997)
    Springer
    Plant molecular biology 33 (1997), S. 467-481 
    Details
    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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    Differential induction of seven 1-aminocyclopropane-1-carboxylate synthase genes by elicitor in suspension cultures of tomato (Lycopersicon esculentum) (1997)
    Springer
    Plant molecular biology 34 (1997), S. 275-286 
    Details
    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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    URL: http://dx.doi.org/10.1023/A:1005800511372
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    Transcription of genes for conglutin γ and a leginsulin-like protein in narrow-leafed lupin (1997)
    Springer
    Plant molecular biology 34 (1997), S. 613-627 
    Details
    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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    An endo-1,4-β-glucanase expressed at high levels in rapidly expanding tissues (1997)
    Springer
    Plant molecular biology 33 (1997), S. 87-95 
    Details
    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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    The wheat protein kinase gene, TaPK3/, of the PKABA1 subfamily is differentially regulated in greening wheat seedlings (1997)
    Springer
    Plant molecular biology 33 (1997), S. 935-941 
    Details
    ISSN: 1573-5028
    Keywords: cold temperature ; dehydration ; greening ; protein kinase ; Triticum aestivum/ ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have identified a new wheat PKABA1/-like protein kinase gene, TaPK3/, that is expressed in greening wheat seedlings. TaPK3 has high sequence homology (97% similarity with some sequence diversity at the 3' end) to the wheat PKABA1 protein kinase mRNA, which is upregulated by cold-temperature treatment, dehydration and abscisic acid (ABA). Use of a TaPK3 gene-specific probe has revealed that TaPK3 is differentially expressed with respect to PKABA1. TaPK3 mRNA accumulates in greening shoot tissue of wheat, but is not affected by dehydration, cold-temperature treatment or ABA. Based on sequence and expression differences, we conclude that expression of the PKABA1/-like protein kinases is not limited to stress responses.
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    Potato SNF1-related protein kinase: molecular cloning, expression analysis and peptide kinase activity measurements (1997)
    Springer
    Plant molecular biology 34 (1997), S. 31-43 
    Details
    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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    Isolation and transcript analysis of gibberellin 20-oxidase genes in pea and bean in relation to fruit development (1997)
    Springer
    Plant molecular biology 33 (1997), S. 1073-1084 
    Details
    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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    Cloning and characterization of tomato leaf senescence-related cDNAs (1997)
    Springer
    Plant molecular biology 33 (1997), S. 641-651 
    Details
    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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    Characterization and expression of two members of the S-adenosylmethionine decarboxylase gene family in carnation flower (1997)
    Springer
    Plant molecular biology 34 (1997), S. 371-382 
    Details
    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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    URL: http://dx.doi.org/10.1023/A:1005811229988
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    Expression of arginine decarboxylase in seedlings of indica rice (Oryza sativa L.) cultivars as affected by salinity stress (1997)
    Springer
    Plant molecular biology 34 (1997), S. 477-483 
    Details
    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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    URL: http://dx.doi.org/10.1023/A:1005802320672
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    cDNA structure and expression patterns of a low-temperature-specific wheat gene tacr7 (1997)
    Springer
    Plant molecular biology 34 (1997), S. 643-650 
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    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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    URL: http://dx.doi.org/10.1023/A:1005852703506
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    Engineered resistance to tomato spotted wilt virus in transgenic peanut expressing the viral nucleocapsid gene (1997)
    Springer
    Transgenic research 6 (1997), S. 297-305 
    Details
    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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    URL: http://dx.doi.org/10.1023/A:1018462729127
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    Rescue of an MMTV transgene by co-integration reveals novel locus control properties of the ovine β-lactoglobulin gene that confer locus commitment to heterogeneous tissues (1998)
    Springer
    Transgenic research 7 (1998), S. 205-212 
    Details
    ISSN: 1573-9368
    Keywords: transgenic mice ; gene expression ; mammary gland ; co-injection ; milk protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In an attempt to enhance the frequency and level of expression of a poor-performing MMTV-driven transgene, we co-integrated this construct with the ovine β-lactoglobulin (BLG) gene in transgenic mice. Seven lines of transgenic mice possessing co-integrated BLG and MMTV-RZ5 transgenes were compared with 12 lines of mice that possessed only the MMTV-RZ5 construct. Co-integration enhanced the frequency of expression in the mammary gland from two out of 12 lines for the MMTV-RZ5 transgene alone, to five out of seven when co-integrated with BLG. Surprisingly, co-integration also resulted in co-expression of the two transgenes in the salivary gland, lung and spleen in addition to the mammary gland. Furthermore, both transgenes were expressed in virgin animals, and throughout pregnancy and lactation, suggesting that the developmental regulation of the locus follows that of the MMTV-promoter. These findings represent a novel locus control property of the ovine BLG gene that confer s commitment of the locus to the mammary gland, but also to a range of heterogeneous tissues possibly defined by the second promoter at the locus
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    URL: http://dx.doi.org/10.1023/A:1008893030461
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    Expression of Human Erythropoietin Transgenes and of the Endogenous Wap Gene in the Mammary Gland of Transgenic Rabbits during Gestation and Lactation (1998)
    Springer
    Transgenic research 7 (1998), S. 311-317 
    Details
    ISSN: 1573-9368
    Keywords: transgenic rabbits ; gene expression ; mammary gland ; erythropoietin ; WAP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An understanding of the expression of transgenes in the mammary gland during gestation and lactation is crucial for the use of transgenic mammals as bioreactors. Here we describe the temporal pattern of expression of the endogenous rabbit WAP gene and human erythropoietin (hEPO) transgenes under the control of rabbit WAP promoter and 3′ flanking sequences. The endogenous rabbit WAP gene was expressed throughout gestation including the day of mating, as well as during lactation in transgenic rabbits bearing a minigene construct. In non-pregnant cycling females, WAP expression was found independent of transgenic status; however, WAP expression was not detected in non-cycling females. The significance of this new finding is not clear at present. hEPO mRNA was detected in mammary gland biopsies from pregnant transgenic rabbits only on day 28 of gestation. During lactation, transcripts were present in mammary gland biopsy samples taken on days 0, 7, 14 and 21. A sharp decline in the levels ...
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    Recombinant human extracellular superoxide dismutase produced in milk of transgenic rabbits (1997)
    Springer
    Transgenic research 6 (1997), S. 271-278 
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    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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    URL: http://dx.doi.org/10.1023/A:1018406611380
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    Lanceispora amphibia gen. et sp. nov., a new amphisphaeriaceous ascomycete inhabiting senescent and fallen leaves of mangrove (1997)
    Springer
    Mycoscience 38 (1997), S. 207-213 
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    ISSN: 1618-2545
    Keywords: Bruguiera gymnorrhiza ; ecology ; Lanceispora amphibia ; mangrove ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Lanceispora amphibia gen. et sp. nov. in the Amphisphaeriaceae is described from senescent and fallen leaves ofBruguiera gymnorrhiza in mangrove forests in the Southwest Islands, Japan. The fungus produces immersed ascomata in leaf tissue, cylindrical asci with an apical ring staining blue with iodine, and oblanceolate ascospores with a septum above the middle. Studies on the fungal succession on the mangrove leaves revealed thatL. amphibia infects senescent leaves on the tree and inhabits intertidal fallen leaves, showing the highest frequency of occurrence at the late stage of decomposition. In culture the optimal conditions for hyphal growth were 20 ppt salinity and 30°C, and those for sexual reproduction were 10 ppt salinity and 25°C. Growth at 0 ppt (fresh water) was depressed. The fungus has amphibious habits, growing on the tree and in intertidal water; and it is adapted to the high osmotic conditions in leaf tissues of the mangrove tree and to the subtropical, brackish water environment of mangrove forests.
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    URL: http://dx.doi.org/10.1007/BF02460855
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    Taxonomical studies on ovariicolous ustomycetes on caryophyllaceae I.Ustilago jehudana andUstilago moenchiae-manticae (1997)
    Springer
    Mycoscience 38 (1997), S. 323-328 
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    ISSN: 1618-2545
    Keywords: Bauhinus ; Microbotryum ; taxonomy ; Ustilago ; ustomycetes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A study of the type specimen ofUstilago jehudana resulted in the correction of the diagnosis. The sori are localized in the host gynoecium but not in the anthers. Morphological characters of the sori and ustospores of the later describedU. moenchiae-manticae are identical with these ofU. jehudana. Ustilago moenchiae-manticae is reduced here to a synonym ofU. jehudana. The smut is reported as new to Bulgaria on a new host, viz.,Moenchia erecta. A new combination,Bauhinus jehudanus, is proposed.
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    Morphology and molecular phylogeny ofTretopileus sphaerophorus, a synnematous hyphomycete with basidiomycetous affinities (1998)
    Springer
    Mycoscience 39 (1998), S. 21-30 
    Details
    ISSN: 1618-2545
    Keywords: Aphyllophorales ; ribosomalDNA ; synnematous hyphomycete ; taxonomy ; Tretopileus sphaerophorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tretopileus sphaerophorus, a synnematous hyphomycete with basidiomycetous affinities was newly isolated from the decaying petiole and peduncle ofCocos nucifera collected in Depok, Indonesia. The species produced first a bulbil as a propagule on the top of a synnema. After the bulbil had fallen, the synnema proliferated about seven times to produce new bulbils, each time making conspicuous nodes at the upper part. By careful morphological observation, clamp connections were confirmed on the hyphae in the specimens and culture. In culture, each hyphal cell with or without a clamp was found to be dikaryotic by DAPI nuclear staining. Germination of the bulbils occurred first from projecting hyphal tips on their upper surface, which have been treated as germ pores. The inner structure of the bulbils, the hyaline mucus of the bulbils, and conidium-like hyphal fragments were also examined. Phylogenetically,T. sphaerophorus was inferred to be related to the Aphyllophorales based on the nuclear encoded small subunit (18S) rDNA using the homology search system (FASTA) and the neighbour-joining method.
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    The order phyllachorales: Taxonomic review (1998)
    Springer
    Mycoscience 39 (1998), S. 97-104 
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    ISSN: 1618-2545
    Keywords: Loculoascomycetes ; phyllachoraceae ; phyllachorales ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The order Phyllachorales contains ascomycetous fungi of considerable economic importance. The group is represented mostly by foliar parasites which produce perithecia under a clypeus, inside a stroma, or do not produce any stromatic tissue. A major taxonomic problem with this order is the lack of reliable morphological characters that clearly delimit the entire group. The main purpose of this review is to provide a clear picture of the taxonomic relationships of the order Phyllachorales, along with a key to the most important genera in the family Phyllachoraceae.
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    A synopsis of the genusScleromitrula (=Verpatinia) (Ascomycotina: Helotiales: Sclerotiniaceae) (1997)
    Springer
    Mycoscience 38 (1997), S. 55-69 
    Details
    ISSN: 1618-2545
    Keywords: discomycetes ; ITS rDNA phylogeny ; morphology ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The systematics ofScleromitrula andVerpatinia of the family Sclerotiniaceae is reevaluated on the basis of morphological, cultural and molecular criteria.Scleromitrula shiraiana, Verpatinia species andCiborinia candolleana share gross morphological, microanatomical and cultural features which clearly distinguish them from the closely relatedCiborinia andRutstroemia species. Phylogenetic analyses of sequences from the internal transcribed spacer region (ITS1, ITS2, and the 5.8S gene) of nuclear ribosomal DNA demonstrate that the stipitate-capitate specimens ofScleromitrula andVerpatinia species plus the stipitate-cupulateCiborinia candolleana constitute a monophyletic clade separate from a clade including the type species ofCiborinia. Scleromitrula is emended to includeS. shiraiana, the new speciesS. rubicola, C. candolleana, and specimens formerly assigned toVerpatinia. A key to the accepted species ofScleromitrula is provided.
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    Ypsilonidium bananisporum sp. nov. (Ceratobasidiales) from Iriomote Island, Japan (1997)
    Springer
    Mycoscience 38 (1997), S. 71-73 
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    ISSN: 1618-2545
    Keywords: Ceratobasidiaceae ; Japan ; taxonomy ; Ypsilonidium bananisporum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ypsilonidium bananisporum sp. nov. belonging to Ceratobasidiales is described and illustrated. This fungus has all the characteristics of the genusYpsilonidium including reticulate-hypochnoid basidiomes, broad hyphae branching at right angles, broadly clavate basidia with two sterigmata, and basidiospores germinating by repetition. It differs from all hitherto known species in the genus by producing suballantoid to banana-shaped basidiospores, measuring 19.5–22×5.5–6 μm.
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    Myxomycetes from Israel IV (1997)
    Springer
    Mycoscience 38 (1997), S. 87-89 
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    ISSN: 1618-2545
    Keywords: Israel ; Mycomycetes ; Physarales ; Stemonitales ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ten taxa of myxomycetes growing mainly withEucalyptus, oak and pine are described. They were found in Upper Galilee, Mt. Carmel and Central parts of the country and all are new to Israel.
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    Mycoleptodiscus terrestris from black pepper roots in the Dominican Republic (1997)
    Springer
    Mycoscience 38 (1997), S. 91-94 
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    ISSN: 1618-2545
    Keywords: hyphomycetes ; identification ; taxonomy ; Tuberculariaceae ; Tuberculariales
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mycoleptodiscus terrestris from black pepper roots in the Dominican Republic is described together with some notes and photomicrographs.
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    Microsphaeropsis rugospora, a new species from Japanese soil (1997)
    Springer
    Mycoscience 38 (1997), S. 429-431 
    Details
    ISSN: 1618-2545
    Keywords: coelomycete ; Japan ; Microsphaeropsis rugospora ; soil fungus ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Abstract In an exploratory survey of soil-borne mitosporic fungi as producers of secondary metabolites useful to the pharmaceutical industry, a pale yellow pycnidial coelomycete was encountered and isolated on potato-dextrose agar. The fungus was characterized as follows: rapid growth on common media, conidiomata which are non-pulvinate, semi-immersed to immersed, nearly globose, glabrous, with a slightly papillate ostiole; pale yellowish brown, translucent, membranaceous peridium; discrete, ampulliform, monophialidic conidiogenous cells; and onecelled, dark brown, globose, thick-walled, rugose conidia which germinate very easily. In accordance with this profile, our isolate is included in the genusMicrosphaeropsis Höhnel. (Morgan-Jones, 1974a, b; Sutton, 1977, 1980; Morgan-Jones and White, 1987; Heiny et al., 1992; Katumoto, 1992). However, it proved to be sufficiently different from all described species ofMicrosphaeropsis to warrant its description as a new species.
    Notes: Abstract A new species ofMicrosphaeropsis (Sphaeropsidales, Coelomycetes),M. rugospora, is described and illustrated. This fungus is characterized by its rapid growth on common media such as oatmeal and potato-carrot agars; semi-immersed to immersed, nearly globose, papillate pycnidias; pale yellowish brown, translucent, membranaceous peridium; monophialidic, ampulliform conidiogenous cells; and one-celled, dark brown, globose conidia ornamented with distinct tubercles. The holotype was isolated from the cultivated soil in Tanegashima Island, southern Japan.
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    On some species ofMycogloea (1998)
    Springer
    Mycoscience 39 (1998), S. 31-36 
    Details
    ISSN: 1618-2545
    Keywords: Mycogloea ; Platygloea ; Platygloeaceae ; Platygloeales ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three new species ofMycogloea are described and illustrated; they are:M. amethystina from Canada,M. nipponica, from Japan, andM. bullata from Thailand.Mycogloea tahitiensis is reported from Japan and additional undescribed taxa in the genus are briefly noted. Some characteristics of the genus are discussed, and a key is provided for six species recognized at this time.
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    Different levels of morphometric variation in three heterogamous dogrose species (Rosa sect.Caninae, Rosaceae) (1997)
    Springer
    Plant systematics and evolution 204 (1997), S. 207-224 
    Details
    ISSN: 1615-6110
    Keywords: Rosaceae ; Rosa sect.Caninae ; Systematics ; taxonomy ; genetic variation ; hemisexual ; apomixis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Difficulties in delimiting well-defined entities in the dogroses (Rosa sect.Caninae) has resulted in very variable taxonomic treatments. The present study was undertaken to provide a background for taxonomy as well as plant breeding. Morphometric diversity was analysed on seedlings obtained from field collections in South Sweden of three species,Rosa dumalis, R. rubiginosa andR. villosa. A canonical variates analysis showed that the three species are relatively distinct whereas two subspecies ofR. dumalis were less well discriminated. Analyses of variance demonstrated that intraspecific variation is pronounced inR. dumalis and, to a lesser extent, inR. villosa.
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    URL: http://dx.doi.org/10.1007/BF00989206
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    Anchusella, a new genus ofBoraginaceae from the Central-Eastern Mediterranean (1997)
    Springer
    Plant systematics and evolution 205 (1997), S. 241-264 
    Details
    ISSN: 1615-6110
    Keywords: Boraginaceae ; Boragineae ; Anchusella ; A. variegata ; A. cretica ; Lycopsis ; Anchusa ; Mediterranean flora ; macromorphology ; micromorphology ; karyology ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The two closely related speciesLycopsis variegata andAnchusa cretica, formerly placed inAnchusa subg.Rivinia, were compared with the type species ofLycopsis andAnchusa, on the basis of a set of macro and microcharacters. The presence of only two fertile stamens as well as other peculiar characters in flower structure, androecium, gynoecium, pollen and fruit, supports the institution of the new genusAnchusella, consisting ofA. variegata andA. cretica. Karyological and eco-chorological aspects are consistent with morphological data in pointing to the autonomy of this genus, which appears characterized by autapomorphic, advanced traits.
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    Comparative HPLC analysis of seed albumins fromVicia faba andV. kalakhensis (Fabaceae) (1997)
    Springer
    Plant systematics and evolution 208 (1997), S. 1-9 
    Details
    ISSN: 1615-6110
    Keywords: Fabaceae ; Vicia faba ; V. kalakhensis ; Seed albumins ; HPLC ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previously reported electrophoretic seed albumin data have shown an unexpected association ofVicia faba withV. kalakhensis. In the present work, seed albumins ofV. faba (subsp.paucijuga and subsp.faba) were compared with those ofV. kalakhensis using ionexchange (IE) and reversed-phase (RP) high-performance liquid chromatography (HPLC). Two subspecies ofV. faba displayed similar seed albumin profiles. On the other hand, seed albumin profiles ofV. faba andV. kalakhensis showed no major protein peak in common either in IE-HPLC or RP-HPLC chromatograms. The reported differences in seed albumin composition ofV. faba andV. kalakhensis are consistent with other taxonomical data showingV. faba to be genetically distant from the wild relatives.
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    URL: http://dx.doi.org/10.1007/BF00986078
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