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  • gene expression  (502)
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  • 1
    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.
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  • 2
    ISSN: 1572-9397
    Keywords: evolutionary algorithms ; genetic algorithms ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Mathematics
    Notes: Abstract This paper shows how evolutionary algorithms can be described in a concise, yet comprehensive and accurate way. A classification scheme is introduced and presented in a tabular form called TEA (Table of Evolutionary Algorithms). It distinguishes between different classes of evolutionary algorithms (e.g., genetic algorithms, ant systems) by enumerating the fundamental ingredients of each of these algorithms. At the end, possible uses of the TEA are illustrated on classical evolutionary algorithms.
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  • 3
    ISSN: 1420-9071
    Keywords: Aldehyde dehydrogenase ; developmental changes ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Previous reports suggested that the major cytosolic aldehyde dehydrogenase (ALDH1) was present in fetal and infant livers, but the major mitochondrial isozyme (ALDH2) was absent or severely diminished. Re-examination by means of starch gel electrophoresis followed by enzyme activity staining, and by means of dot blot immuno-hybridization of liver samples with known genotypes of theALDH 2 locus, indicated that bothALDH 1 andALDH 2 genes are expressed in fetal and infant livers. In addition, ALDH4 isozyme was also observed. The results imply that a fetus with the ‘usual’ homozygousALDH 2 1 /ALDH 2 1 genotype, but not one with the atypicalALDH 2 1 /ALDH 2 2 orALDH 2 2 /ALDH 2 2 , is capable of detoxifying acetaldehyde transferred from the mother.
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  • 4
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    Cellular and molecular life sciences 47 (1991), S. 321-331 
    ISSN: 1420-9071
    Keywords: Symbiosis ; ectomycorrhiza ; ectomycorrhiza development ; gene expression ; ectomycorrhizins ; protein patterns
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An ectomycorrhiza, a specialized root organ, is the result of a complex interaction leading to a finely-tuned symbiosis between a plant and a compatible ectomycorrhizal fungus. Ultrastructural observations combined with cytochemical and biochemical studies reveal that structural and metabolic changes in the symbiont cells lead to the final phenotype of the active ectomycorrhiza. In the present review these changes are interpreted as changes in gene expression and discussed within the context of ectomycorrhiza development. Recent genetic data indicate that the continued vegetative growth of the ectomycorrhizal hyphae and the root tissues, and their ability to switch to symbiotic organ formation, is basically controlled by developmentally critical genes. The activity of these ‘symbiotic genes’ during the differentiation of ectomycorrhizas is associated with extensive changes in the concentration of particular polypeptides and protein biosynthesis. The present state of knowledge about the developmental biology of ectomycorrhizas allows only speculation about the events during their development.
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  • 5
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    Cellular and molecular life sciences 49 (1993), S. 133-137 
    ISSN: 1420-9071
    Keywords: Fetal hemoglobin ; sickle cell anemia ; β thalassemia ; butyrate ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The inherited β-hemoglobinopathies (sickle cell disease and β thalassemia) are the result of a mutation in the adult (β) globin gene. The fetal globin chain, encoded by the γ globin genes, can substitute for the mutated or defective β globin chain, but expression of the γ globin gene is developmentally inactivated prior to birth. Reinducing expression of the normal fetal globin genes is a preferred method of ameliorating sickle cell disease and the β thalassemias. Stimulation of as little as 4–8% fetal globin synthesis in the bone marrow can produce 〉20% fetal hemoglobin in the peripheral circulation, due to enhanced survival of red blood cells containing both sickle and fetal hemoglobin, compared to those containing sickle hemoglobin alone. Butyric acid and butyrate derivatives are generally safe compounds which induce fetal hemoglobin production by stimulating the promoter of the fetal globin genes. An initial trial with the parent compound, delivered as Arginine Butyrate, has demonstrated rapid stimulation of fetal globin expression to levels that have been shown to ameliorate these conditions. Phase 1 trials of an oral butyrate derivative with a long plasma half-life have just begun. These agents now provide a specific new apporach for ameliorating these classic molecular disorders and merit further investigation in larger patient populations.
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  • 6
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    Cellular and molecular life sciences 52 (1996), S. 942-949 
    ISSN: 1420-9071
    Keywords: Apoptosis ; transglutaminase ; signalling ; gene expression ; promoter elements ; retinoic acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Clarification of the molecular details of forms of natural cell death, including apoptosis, has become one of the most challenging issues of contemporary biomedical sciences. One of the effector elements of various cell death pathways is the covalent cross-linking of cellular proteins by transglutaminases. This review will discuss the accumulating data related to the induction and regulation of these enzymes, particularly of tissue type transglutaminase, in the molecular program of cell death. A wide range of signalling pathways can lead to the parallel induction of apoptosis and transglutaminase, providing a handle for better understanding the exact molecular interactions responsible for the mechanism of regulated cell death.
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  • 7
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    Cellular and molecular life sciences 51 (1995), S. 1116-1123 
    ISSN: 1420-9071
    Keywords: Antisense RNA ; gene expression ; insertional mutagenesis ; physical mapping ; reporter genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Over the past ten years, powerful molecular genetic techniques have been developed to analyze gene function inDictyostelium. DNA-mediated transformation using a variety of selections and vectors has allowed the introduction of wild-type or modified genes that are under various forms of transcriptional control. Homologous recombination is efficient and can be used to modify the genome in precise ways. In addition, it is now possible to clone genes based on their mutant phenotype alone, either by insertional mutagenesis, or by screening antisense expression cDNA libraries. Finally, a nearly complete physical map of the genome is available and so genes are easily mapped by physical techniques. We discuss many of these advances within the context of major research problems presently under study.
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  • 8
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    Cellular and molecular life sciences 50 (1994), S. 958-962 
    ISSN: 1420-9071
    Keywords: Plasminogen activator ; active oxygen ; gene expression ; radical scavengers ; endothelial cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Active oxygen, produced by cultured cells following stimulation with various growth factors, seems to be involved in signal transduction leading to cellular responses such as gene expression and growth modulation. In the present study, the intracellular oxidation state was measured in immortalized human endothelial cells (ECV304) after treatment with tumor necrosis factor (TNF)α, using a fluorescent dye and a laser-scanning confocal microscope. The intracellular oxidation state was increased 60 min after the addition of TNFα, and this increase was abolished by a radical scavenger, N-acetylcysteine (NAC), which is also a precursor of glutathione, and by pyrrolidine dithiocarbamate (PDTC). TNFα increased the steady state level of urokinase-type plasminogen activator (uPA), and NAC inhibited this increase at a dose that also inhibited the increase in the intracellular oxidation state. PDTC, on the other hand, did not affect the induction of the uPA gene by TNFα. These results suggest that intracellular glutathione level rather than the oxidation state is necessary for the induction of the uPA gene by TNFα.
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  • 9
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    Cellular and molecular life sciences 49 (1993), S. 447-455 
    ISSN: 1420-9071
    Keywords: Proteoglycan ; chondroitin sulfate ; decorin ; gene expression ; tumor stroma ; DNA methylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Tumor stroma is a specialized form of tissue that is associated with epithelial neoplasms. Recent evidence indicates that significant changes in proteoglycan content occur in the tumor stroma and that these alterations could support tumor progression and invasion as well as tumor growth. Our main hypothesis is that the generation of tumor stroma is under direct control of the neoplastic cells and that, via a feedback loop, altered proteoglycan gene expression would influence the behavior of tumor cells. In this review, we will focus primarily on the work from our laboratory related to the altered expression of chondroitin sulfate proteoglycan and its role in tumor development and progression. The connective tissue stroma of human colon cancer is enriched in chondroitin sulfate and the stromal cell elements, primarily colon fibroblasts and smooth muscle cells, are responsible for this biosynthetic increase. These changes can be reproduced in vitro by using either tumor metabolites or co-cultures of human colon carcinoma cells and colon mesenchymal cells. The levels of decorin, a leucine-rich proteoglycan involved in the regulation of matrix assembly and cell proliferation, are markedly elevated in the stroma of colon carcinoma. These changes correlate with a marked increase in decorin mRNA levels and a concurrent hypomethylation of decorin gene, a DNA alteration associated with enhanced gene expression. Elucidation of decorin gene structure has revealed an unexpected degree of complexity in the 5′ untranslated region of the gene with two leader exons that are alternatively spliced to the second coding exon. Furthermore, a transforming growth factor beta (TGF-β)-negative element is present in the promoter region of decorin gene. This regulatory domain is likely to be implicated in the silencing of decorin gene by TGF-β and may contribute to the regulation of this matrix gene in the tumor stroma.
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  • 10
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    Cellular and molecular life sciences 51 (1995), S. 606-611 
    ISSN: 1420-9071
    Keywords: Metallothionein ; isometallothioneins ; gene expression ; rabbit kidney cell-line ; cadmium adaptation ; zinc adaptation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We explored the molecular genetics underlying the massive induction of isoMTs by Zn2+ or Cd2+ in metal tolerant rabbit kidney (RK-13) sub-line cells, using band shift assays and Southern blotting analysis. In sub-line cells accommodated to intermediate metal concentrations (100 μM Zn2+; 1–20 μM Cd2+) evidence suggested that the increase in the capacity for isoMT synthesis is brought about by an increased binding activity of the nuclear transcription factors MTF-1 and Sp1. Using quantitative band shift analysis with a mouse MRE-d oligonucleotide probe, the binding of both transcription factors was found to be enhanced two to three times over the binding activity measured in the unexposed parental RK-13 cells. Their increase in binding activity is probably the cause of the overexpression of MT genes and the development of metal tolerance in these cells. In cells tolerant to the highest concentrations of metal the analysis of Southern blot signals revealed MT gene amplification to be the most probable cause of the increased MT production. Thus, in cells of sub-lines growing in the presence of 350 μM Zn2+, two of the isoMT genes were coordinately triplicated and in cells tolerant to 150 μM Cd2+ one isoMT gene was amplified two-fold.
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  • 11
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    BioMetals 11 (1998), S. 345-358 
    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
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  • 12
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    Plant molecular biology reporter 16 (1998), S. 323-339 
    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.
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  • 13
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    Plant molecular biology reporter 17 (1999), S. 371-383 
    ISSN: 1572-9818
    Keywords: epidermal peel ; extraction ; gene expression ; stomata ; tree tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Stomatal guard cells are critical for maintenance of plant homeostasis and represent an interesting cell type for studies of leaf cell differentiation and patterning. Here we describe techniques for the isolation of guard cell RNA and protein from blended epidermal peels of Nicotiana glauca. The RNA isolation procedure is a modification of the hot borate method, which is particularly well-suited for recalcitrant tissues. Protein was extracted by disrupting guard cell-enriched epidermis with a French® press. This system offers the following advantages: relatively high yield, low or no contamination by other cell types, fresh tissue as a source of RNA and protein rather than protoplasts, and a plant species that is readily transformable. These techniques will allow for cloning and analysis of genes expressed in guard cells, application of traditional biochemical techniques to guard cell proteins, as well as characterization of genetic manipulation of guard cell function in transgenic plants.
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  • 14
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    Journal of paleolimnology 9 (1993), S. 63-68 
    ISSN: 1573-0417
    Keywords: Chrysophyceae ; stomatocysts ; membrane topography ; biogenic silica deposition ; surface pattern ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
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  • 15
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    Journal of paleolimnology 6 (1991), S. 257-260 
    ISSN: 1573-0417
    Keywords: Chrysophyceae ; stomatocysts ; biogenic silica deposition ; polymer gels ; surface pattern ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract The utility of classifying chrysophyte stomatocysts by their characteristic honeycomb and ridge patterns is questioned, because a strikingly similar expanding pattern appears on the surface of ionized polymer gels during osmotical swelling as a result of simple physical forces. The rapid accumulation of silicate into a spherical cyst inside a chrysophyte cell appears to be as a physical process sufficiently similar to result in an analogous pattern in microscopic scale. Chrysophyte stomatocysts that possess honeycomb or ridge patterns could be regarded as ‘frozen moments’ of the pattern evolution during the silicate gel phase. As a consequence, such structures should have little taxonomical value.
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  • 16
    ISSN: 1573-0832
    Keywords: DNA ; Exophiala dermatitidis ; Exophiala gougerotii ; Exophiala jeanselmei ; mitochondria ; restriction profiles ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Mitochondrial DNA (mtDNA) analysis with Hae III, Hind. III and Msp I was performed in 45 Exophiala jeanselmei strains (30 Phialophora jeanselmei and 15 Phialophora gougerotii strains) and 31 Exophiala dermatitidis strains. The results were as follows, 1) P. jeanselmei and P. gougerotii are identical, 2) E. jeanselmei is classified into 18 types based on restriction profiles, 3) two strains of E. jeanselmei CBS 577.76 and CBS 578.76 are identified as E. dermatitidis, 4) E. dermatitidis has no intraspecific variation and is definitely distinct from E. jeanselmei, 5) E. jeanselmei is suggested to be a complex organism because of extensive mtDNA polymorphism.
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  • 17
    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.
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  • 18
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    Antonie van Leeuwenhoek 57 (1990), S. 223-236 
    ISSN: 1572-9699
    Keywords: fluorescent Pseudomonas ; root associated ; siderophores ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The phenetic taxonomy of 110 fluorescent bacterial strains, isolated from the roots of tomatoes and other plants was numerically studied through 97 features including 69 assimilation tests. Thirty-two reference strains of various Pseudomonas spp. were additionally included. The strains clustered into 16 clusters at the 74% similarity level when using Jaccard similarity coefficients. Almost all field strains belonged to the P. fluorescens/P. putida-complex while none clustered with P. syringae and allied bacteria. The biovar II branch, as well as the newly described biovar VI of P. fluorescens, made up 55% and 20% respectively, of the field strains; two % were allocated to P. fluorescens biovar I and three % to biovar IV. Eleven % of the root associated strains were designated P. putida; six strains were biovar A, three strains biovar B while four strains could not be referred to any known biovar. The continuum within the P. fluorescens/P. putida-complex as well as the taxonomic status of the six biovars of P. fluorescens and the three biovars of P. putida are discussed.
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  • 19
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    Antonie van Leeuwenhoek 58 (1990), S. 95-98 
    ISSN: 1572-9699
    Keywords: Zygozyma smithiae ; Lipomycetaceae ; ambrosia yeasts ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A new species of the genus Zygozyma, Z. smithiae, was recovered from frass of the ambrosia beetle, Crossotarsus externedentatus in Northern Natal. A description of the new species and key to the genus are given.
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  • 20
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    Antonie van Leeuwenhoek 57 (1990), S. 59-61 
    ISSN: 1572-9699
    Keywords: Arxula ; Candidaceae ; yeasts ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The new genus Arxula is proposed for the classification of xerotolerant, ascomycetous, anamorphic, arthroconidial yeasts. The genus is considered to be of endomycetaceous affinity.
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  • 21
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    Antonie van Leeuwenhoek 59 (1991), S. 65-76 
    ISSN: 1572-9699
    Keywords: anaerobic respiration ; FNR protein ; oxygen regulation ; gene expression ; E. coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Molecular oxygen is an important regulatory signal in facultative anaerobic bacteria and controles the expression of a great variety of genes positively or negatively. The expression of anaerobic respiration and of related functions of E. coli is controlled by the positive gene regulator FNR, which activates transcription in the absence of O2. The regulated genes carry a FNR consensus sequence upstream of the promoter. Under the same conditions FNR represses some of the genes of aerobic respiration. The binding to the DNA occurs by an α-helix-turn-α-helix DNA-binding domain. FNR contains 5 cysteine residues, four of which are arranged in a cluster close to the N-terminal end. For the function of FNR as a O2-dependent regulator three of the cysteine residues in the cluster and the residue outside the cluster are essential. FNR binds iron as a cofactor which most likely is involved in the O2-sensing by the protein. The experiments indicate that the cysteine residues are responsible for the binding of the iron. From the protein in vivo two functional states can be differentiated, an aerobic or metal-depleted form and an anaerobic form. Only the anaerobic form acts as a gene activator or repressor. Sensing of O2 or of positive redox potentials by the iron ion is thought to cause the conversion of the two functional states. The FNR protein in addition contains a potential nucleotide binding domain. The significance and function of this site is not clear.
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  • 22
    ISSN: 1572-9699
    Keywords: basidiomycetous yeasts ; Kurtzmanomyces tardus ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A new yeast species of basidiomycetous affinity Kurtzmanomyces tardus was isolated from contaminated demineralized water. It differs from K. nectairii, the only other Kurtzmanomyces species so far described, in its carbon assimilation pattern and low DNA-DNA homology (2.3%±2.1).
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  • 23
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    Antonie van Leeuwenhoek 61 (1992), S. 245-248 
    ISSN: 1572-9699
    Keywords: Sporobolomyces lactosus ; yeasts ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A new species of ballistospore-forming yeasts was recovered and its description given.Sporobolomyces lactosus, isolated from activated sludge treating petrochemical wastes, produces pinkish-coral to pink colored colonies, assimilates lactose and has Q-10 as the major ubiquinone.
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  • 24
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    Antonie van Leeuwenhoek 61 (1992), S. 289-316 
    ISSN: 1572-9699
    Keywords: Trichosporon ; white piedra ; systemic mycosis ; taxonomy ; nDNA ; rRNA ; ubiquinone systems ; basidiomycetous yeasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The genusTrichosporon was revised using characters of morphology, ultrastructure, physiology, ubiquinone systems, mol% G+C of DNA, DNA/DNA reassociations and 26S ribosomal RNA partial sequences. A total of 101 strains was used, including all available type and authentic cultures of previously described taxa. Nineteen taxa could be distinguished, 15 of which having Q-9 coenzyme systems and 4 having Q-10. Sixteen previously described names were reduced to synonymy. One new species was described. The genus is characterized by the presence of arthroconidia. Few species possess further diagnostic morphological characters, such as the presence of appressoria, macroconidia or meristematic conidiation. The septa of two species were found to be non-perforate, while those of the remaining species contained dolipores at variable degrees of differentiation, with or without vesicular or tubular parenthesomes. All species were able to assimilate a large number of carbon compounds; visible CO2 production was absent. The genus was found to be fairly homogeneous on the basis of a phylogenetic analysis of partial 26S rRNA sequences, with the exception ofT. pullulans which proved to be unrelated. Most taxa were found to occupy well-defined ecological niches. Within the group of taxa isolated from humans, a distinction found be made between those involved in systemic mycoses and those which mainly caused public or non-public white piedras, respectively. One species was consistently associated with animals, while others came mainly from soil or water. One species was mesophilic and another psychrophilic.
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  • 25
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    Antonie van Leeuwenhoek 62 (1992), S. 131-153 
    ISSN: 1572-9699
    Keywords: mitochondrial DNA ; mutational analysis ; nucleo-mitochondrial interactions ; gene expression ; membrane assembly ; respiratory deficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In contrast to most other organisms, the yeastSaccharomyces cerevisiae can survive without functional mitochondria. This ability has been exploited in genetic approaches to the study of mitochondrial biogenesis. In the last two decades, mitochondrial genetics have made major contributions to the identification of genes on the mitochondrial genome, the mapping of these genes and the establishment of structure-function relationships in the products they encode. In parallel, more than 200 complementation groups, corresponding to as many nuclear genes necessary for mitochondrial function or biogenesis have been described. Many of the latter are required for post-transcriptional events in mitochondrial gene expression, including the processing of mitochondrial pre-RNAs, the translation of mitochondrial mRNAs, or the assembly of mitochondrial translation products into the membrane. The aim of this review is to describe the genetic approaches used to unravel the intricacies of mitochondrial biogenesis and to summarize recent insights gained from their application.
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  • 26
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    Antonie van Leeuwenhoek 71 (1997), S. 375-378 
    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.
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  • 27
    ISSN: 1572-9699
    Keywords: phylogeny ; prosthecobacter ; taxonomy ; Verrucomicrobia ; Verrucomicrobiae
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    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.
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  • 28
    ISSN: 1572-9699
    Keywords: keratinophilic fungi ; Neoarachnotheca ; Neoarachnotheca keratinophila ; Nannizziopsis tropicalis ; Onygenales ; taxonomy
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    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.
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  • 29
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    Antonie van Leeuwenhoek 74 (1998), S. 229-235 
    ISSN: 1572-9699
    Keywords: Dipodascus capitatus ; D.spicifer ; Geotrichum clavatum ; yeast ; taxonomy ; DNA heterogeneity
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    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.
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  • 30
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    Antonie van Leeuwenhoek 66 (1994), S. 337-342 
    ISSN: 1572-9699
    Keywords: Debaryomyces ; phylogeny ; ribosomal RNA ; taxonomy ; Wingea ; yeasts
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    Topics: Biology
    Notes: Abstract Extent of divergence in partial nucleotide sequences from large and small subunit ribosomal RNAs was used to estimate the evolutionary relationship between the generaWingea andDebaryomyces. These data showed the monotypic genusWingea to be congeneric withDebaryomyces, and it is proposed to transferW. robertsii toDebaryomyces.
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  • 31
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    Plant molecular biology reporter 16 (1998), S. 147-155 
    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.
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  • 32
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    Journal of insect conservation 2 (1998), S. 151-159 
    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.
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  • 33
    ISSN: 1572-9745
    Keywords: Biological control ; insects ; pathogens ; germination ; taxonomy ; genetic variability
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    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
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  • 34
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    Journal of the history of biology 32 (1999), S. 509-555 
    ISSN: 1573-0387
    Keywords: Louis Agassiz ; Spencer Baird ; Charles Girard ; reputation ; taxonomy ; ichthyology ; Museum of Comparative Zoology ; Smithsonian Institution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , History
    Notes: Abstract The reputations of scientists among their contemporaries depend not only on accomplishment, but also on interactions affected by influence and personality. The historical lore of most fields of scientific endeavor preserve these reputations, often through the identification of founders, innovators, and prolific workers whose contributions are considered fundamental to progress in the field. Historians frequently rely on the historical lore of scientists to guide their studies of the development of ideas, exhibiting justifiable caution in reassessing reputations in the light of current knowledge. However, the transmission of historical lore can obscure the relative importance of accomplishment, influence and personality in shaping contemporary reputations, leaving the historian to either accept reputations at face value or attempt to reconstruct the context in which they were created. The science of taxonomy, because of its rules of priority, leaves a relatively accurate record of historical accomplishment through the persistence of taxa in catalogues and faunal guides. These records allow the modern historian an unbiased means to assess the relative accomplishments of historical figures and therefore a means to critically reassess reputations independent of personality and influence. In the historical lore of North American ichthyology, Louis Agassiz at Harvard and Spencer Baird at the Smithsonian emerge as central figures in the early development of the field during the mid-1800s, contributing not only through the quality and quantity of their science, but also through their roles as institutional leaders and mentors to workers who followed. Charles Girard, originally a student of Agassiz's and later a coworker with Baird, receives little notice in the history of ichthyology, and his reputation is that of a minor player in the initial description of the North American fish fauna, and one whose work appears to have been flawed or even careless when compared to his contemporaries. However, a review of both contemporary and modern taxonomic works reveals that Girard's productivity far exceeded that of either Agassiz or Baird. Furthermore, an examination of the tendency of Girard and his contemporaries to introduce synonymous names into the literature, which might reflect careless or uncritical work, suggests that Girard was among the more accomplished workers of hisera, including Agassiz and Baird. Girard's low ranking in the folklore of North American ichthyology, therefore, can not be attributed to discernible shortcomings in his scientific work, but rather to a public and private campaign of criticism waged by Agassiz after Girard's departure from Harvard. While Agassiz's dispute with Girard stemmed from their personal interactions, he expressed them as criticisms of Girard's work, and thus helped shape Girard's scientific reputation as it has been transmitted through the lore of ichthyology. This case study reveals how scientific reputation may not always rest on accomplishment, but can be influenced by personal interactions obscured by time but nonetheless important to history.
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  • 35
    ISSN: 1420-9098
    Keywords: Thaumatomyrmex ; taxonomy ; comparative morphology ; predation ; Polyxenidae
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    Topics: Biology
    Notes: Summary We describe, for the first time, the predatory behaviour ofThaumatomyrmex ants on millipedes of the family Polyxenidae, based on field observations ofT. atrox and a field and laboratory study ofT. contumax. The capture of the prey and the removal process of its body-covering setae by the ants before they eat the millipede are described. This specialized behaviour in at least two species of the genus, belonging to two distinct groups of species, indicates a general trend inThaumatomyrmex. We coupled this study with a comparative morphological analysis of the mouthparts and digestive tube of these and otherThaumatomyrmex species. Also, we report the first case of sympatry in the genus, which suggests thatThaumatomyrmex includes several species, and not only one highly variable taxon, as hypothetized earlier.
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  • 36
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    Cellular and molecular life sciences 49 (1993), S. 1055-1058 
    ISSN: 1420-9071
    Keywords: Moderate halophiles ; taxonomy ; Salinicoccus roseus ; Marinococcus hispanicus
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    Topics: Biology , Medicine
    Notes: Abstract The Gram-positive moderately halophilic cocci validly described up to date areMicrococcus halobius, Sporosarcina halophila, Marinococcus halophilus, Marinococcus albus, Marinococcus hispanicus, and the recently proposedSalinicoccus roseus. Our chemotaxonomic studies on these bacteria have revealed thatMarinococcus hispanicus should be placed in the genusSalinicoccus. DNA-DNA hybridization studies showed that this species should be considered as a separate species ofS. roseus. Thus, we have proposed transferring it to the genusSalinicoccus, asS. hispanicus. Besides, phenotypic and chemotaxonomic features of the culture collection moderate halophilesMicrococcus sp. strains CCM 168 and CCM 1405 indicate that they should be included in the speciesSalinicoccus roseus. The description of this species was based only on one strain.
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  • 37
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    Cellular and molecular life sciences 52 (1996), S. 888-891 
    ISSN: 1420-9071
    Keywords: Ageing ; rat ; brain ; gene expression ; differential display
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    Topics: Biology , Medicine
    Notes: Abstract We have used the polymerase chain reaction (PCR)-based technique of differential display to analyse changes in gene expression during ageing of the rat brain. In this approach we have compared three young adult (6 months) with three old adult (20 months) animals. RNA preparations from the homogenised brains were subjected to reverse transcriptase (RT)-PCR using 36 different combinations of primer pairs. Any PCR product which was consistently found to be more prominent in the three young brains compared to the three old brains, and vice versa, was scored as potentially representing a gene which was differentially expressed during the ageing of this tissue. Out of a possible 2000+PCR products we identified 44 that might represent genes that exhibit differential expression during ageing of the rat brain. An initial screen of these fragments, by Southern-blotting the PCR products and hybridising them with cDNA probes derived from either young or old brain RNA preparations, indicated that 40% of them represented genes that were differentially expressed. This approach is likely to prove invaluable for identifying cohorts of genes that show differential expression during the ageing process.
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  • 38
    ISSN: 1420-9055
    Keywords: Heterotrophic bacteria ; sediments ; taxonomy ; seasonal variations ; aerobic bacterial population ; anaerobic bacterial population ; Lake Lugano (Lago di Lugano)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This research was carried out with the aim to explore the heterotrophic microbial population of two sediments in different oxic conditions of the Lake of Lugano (Lago di Lugano). The values of the viable bacterial counts found in our sediment samples were typical for an eutrophic lake. The increase in the proportion of anaerobic to aerobic bacteria at 30 °C observed in the sediment samples of Agno may reflect the depletion of oxygen concentrations in the water column. The generaAeromonas, Pseudomonas, Bacillus andClostridium, as well as strains belonging to the Coryneform-group, represented the major taxonomic groups of heterotrophic bacterial communities in the water-sediment interface.
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  • 39
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    Cellular and molecular life sciences 47 (1991), S. 22-31 
    ISSN: 1420-9071
    Keywords: Atherosclerosis ; cellular differentiation ; gene expression ; foam cells ; lipoproteins ; phorbol esters ; transcription factors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary THP-1 is a human monocytic leukemia cell line. After treatment with phorbol esters, THP-1 cells differentiate into macrophage-like cells which mimic native monocyte-derived macrophages in several respects. Compared to other human myeloid cell lines, such as HL-60, U937, KG-1, or HEL cell lines, differentiated THP-1 cells behave more like native monocyte-derived macrophages. Because of these characteristics, the THP-1 cell line provides a valuable model for studying the mechanisms involved in macrophage differentiation, and for exploring the regulation of macrophage-specific genes as they relate to physiological functions displayed by these cells.
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  • 40
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    Cellular and molecular life sciences 47 (1991), S. 905-912 
    ISSN: 1420-9071
    Keywords: Gene transfer ; gene modification ; gene expression ; livestock ; transgenic animal ; pharmaceutical proteins ; milk composition
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    Topics: Biology , Medicine
    Notes: Abstract There is every reason to expect that it will be possible within the next few years to begin to use farm animals to produce large quantities of some of the human proteins that are needed for the treatment of disease. Revolutionary new opportunities for the production of novel proteins in milk have been created by the development of methods for gene transfer. Exploitation of these opportunities depends upon selection and cloning of milk protein genes and identification of the sequences that govern tissue specific hormonally induced expression in the mammary gland. Studies with three genes, ovine β-lactoglobulin, rat β-casein and whey acidic protein of rat and mouse, suggest that they may all meet this requirement. Fragments of the ovine β-lactoglobulin, murine whey acidic protein and rabbit β-casein genes have directed production of novel proteins in the milk of transgenic mice, sheep, rabbits and pigs. The proteins were biologically active and usually co-migrated with authentic proteins. In early experiments, protein concentration was low, but our recent observations suggest that fusion genes containing genomic clones direct production of concentrations of protein that are suitable for commercial exploitation. In the longer term, two approaches may offer the potential of more reliable expression. Control elements capable of directing expression that is independent of site of insertion of the gene, but dependent on the number of copies of the gene, have been identified for a small number of genes. The availability of such elements for the milk protein genes would increase the reliability of gene expression considerably. Alternatively, targeted mutation of genes may allow the insertion of coding sequences within an existing gene so avoiding position effects.
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  • 41
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    Cellular and molecular life sciences 47 (1991), S. 866-877 
    ISSN: 1420-9071
    Keywords: Transgenic mice ; microinjection ; recombinant DNA ; gene expression ; transcription factors ; chromatin ; homologous recombination ; episomal maintenance ; embryonic stem cells ; germ line ; position-effect ; mosaicism ; globin genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This chapter is an attempt to summarize some commonly accepted and some more subjective opinions about the regulation of transgene expression in laboratory animals. After a short historical introduction, I present some general notions regarding gene structure/function. The spotlight shifts then to the description of the most popular techniques for gene transfer, including the targeted gene replacement. The different approaches are briefly discussed in terms of intrinsic advantages and limitations regarding gene expression patterns. Furthermore, the role of enhancers, promoters and othercis-acting elements such as silencers and dominant control regions as well as their involvement in the chromatin on-off state are discussed on the basis of a specific example studied in our laboratory. The review concludes by presenting recent results and the new perspectives opening in the field of ‘surrogate’ (also called ‘reversed’) genetics. Some problems which remain to be solved both at the technical as well as at the social-ethical level are also briefly presented.
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  • 42
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    Plant cell reports 10 (1991), S. 308-314 
    ISSN: 1432-203X
    Keywords: Agrobacterium tumefaciens ; Brassica juncea ; genetic transformation ; gene expression ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An efficient system for gene transfer into plants of Brassica juncea var. India Mustard, mediated by Agrobacterium tumefaciens. was developed through the manipulation of the culture medium and the use of the appropriate Agrobacterium strain. High frequency shoot regeneration (90–100%) was obtained from hypocotyl explants grown on medium containing 0.9% agarose, 3.3 mg/L AgNO3 and 0.5–2 mg/L BA in combination with 0.01–0.05 mg/L 2,4-D or 0.1–1 mg/L NAA. Of all the Agrobacterium strains tested, A. tumefaciens A208-SE, carrying the disarmed Ti plasmid and a binary vector pROA93, was the most effective for B. juncea transformation. pROA93 carries the coding sequences of the NPTII and the GUS genes, both driven by a common CaMV 35S promoter in two divergent directions. Inoculated explants grown on the selection medium in the presence of 0.5 mg/L BA and 0.1 mg/L NAA gave rise to transgenic shoots at the highest frequency (9%). All Ro transgenic plants were phenotypically normal, but variation in expression patterns of the GUS gene occurred among the transgenic plants in an organ- and tissue-specific manner. Both the NPTII and the GUS genes were transmitted to the R1 seed progeny and showed co-segregation.
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  • 43
    ISSN: 1432-203X
    Keywords: glyphosate ; gene expression ; gene amplification ; cell culture ; resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The stability and expression of amplified 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) genes was examined in glyphosate resistant tobacco cells grown in glyphosate-free medium, and in plantlets regenerated from resistant cells. Amplified DNA was maintained in resistant cells grown in the absence of glyphosate for three years. Amplified EPSPS genes were retained in regenerated plantlets at levels comparable to those observed in the resistant cells, and EPSPS mRNA was overexpressed (compared to unselected plantlets). However, glyphosate resistance in cell lines grown in glyphosate-free medium declined 7-fold, and in regenerated plantlets approximately 20-fold, compared to resistant cells maintained under glyphosate selection. In plantlets, reduced resistance correlated with lower levels of EPSPS mRNA. Plantlets regenerated from resistant cells exhibited morphological variation, and had an approximate doubling of their nuclear genome size.
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  • 44
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    Journal of industrial microbiology and biotechnology 17 (1996), S. 328-345 
    ISSN: 1476-5535
    Keywords: hyphomycetes ; ascomycetes ; taxonomy ; ecology ; stream biology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract There are more than 600 species of freshwater fungi with more known from temperate, as compared to tropical regions. These includeca 340 ascomycetes, 300 deuteromycetes, and a number of lower fungi which are not discussed here.Aniptodera, Annulatascus, Massarina, Ophioceras andPseudohalonectria are common freshwater ascomycetes, which appear to be well adapted for this lifestyle either in their ascospore types or their competitive-degradative characters. The most common genera of wood-inhabiting deuteromycetes includeCancellidium, Dactylaria, Dictyosporium andHelicomyces. They are categorized into four groups depending on their form and life style: the ingoldian hyphomycetes; the aero-aquatic hyphomycetes; the terrestrial-aquatic hyphomycetes; and the submerged-aquatic hyphomycetes. The adaptations of aquatic fungi for their dispersal and subsequent attachment to new substrates are discussed.
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  • 45
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    World journal of microbiology and biotechnology 10 (1994), S. 88-92 
    ISSN: 1573-0972
    Keywords: Intelligent screening ; microbial hydroxylation ; multivariate analyses ; taxonomy ; terpenoids
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    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Forty strains of bacteria and 60 of fungi were screened with globulol, 1,8-cineol and cedrol and the strains compared, on the basis of the resulting products, using multivariate analyses. The taxonomic position of the strains is mirrored in their biotransformation capability. Fungi and bacteria, fungal phyla, and Gram-positive and-negative bacteria could all be distinguished in a discriminant analysis using 12 substrates. Hierarchical clustering yielded five groups of strains with distinct biotransformation activity. Such clustering will allow a more efficient screening than before.
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  • 46
    ISSN: 1573-4919
    Keywords: mitochondrial biogenesis ; cytochrome c oxidase ; mRNA quantitation ; gene expression
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary Cytochrome c oxidase (COX), like other multi-subunit components of the respiratory chain, is controlled by both the nuclear and the mitochondrial genome. In order to find wether there is a close relationship between mRNAs encoded by the nucleus and by the mitochondrion, and between these mRNAs and enzyme activity, we compared six rat tissues (ventricle, liver, m. soleus, m. plantaris, and the white and red portions of m. gastrocnemius). We found a tenfold range for COX activity, a tenfold range for the contents of mRNA III (mitochondrial) and mRNA VIc (nuclear), a threefold range for total [poly(A)+] mRNA content and a sevenfold range for total RNA content in these tissues. The ratio of mRNA III to mRNA VIc was equal in each tissue, indicating the presence of a mechanism that coordinates the two genomes. There was a good correlation between mRNA content and COX activity (r = 0.78 for VIc, r = 0.77 for 111; p 〈 0.0001), demonstrating that the expression of this enzyme is mainly under pretranslational control.
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  • 47
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; rat liver
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The involvement of a hypocalcemic hormone calcitonin (CT) in the expression of hepatic Ca2+-binding protein regucalcin mRNA was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb). A single oral administration of calcium chloride (100 mg Ca/100 g body weight) to rats induced a remarkable increase in the serum calcium concentration and a corresponding elevation of the liver calcium content during 120 min after the administration. Thyroparathyroidectomy (TPTX) did not cause a significant increase in the liver calcium content after calcium administration. Hepatic regucalcin mRNA level was markedly elevated by calcium administration; the level was about 180% of controls at 60 min after the administration. This increase was completely abolished by TPTX. A single subcutaneous administration of CT (synthetic eel CT; 25–100 MRC mU/100 g) to TPTX rats received oral administration of calcium (100 mg/100 g) produced a remarkable increase in hepatic regucalcin mRNA levels; the level was about 280% of controls with the dose of 25 MRC mU CT/100 g. The present finding suggests that the expression of hepatic mRNA is stimulated by CT, and that the hormonal effect is mediated through Ca2+ in rat liver.
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  • 48
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; phenobarbital ; rat liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of phenobarbital on the expression of calcium-binding protein regucalcin mRNA in rat liver was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin cDNA (0.9 kb of open reading frame). Phenobarbital (4, 8 and 12 mg/ 100 g body weight) was intraperitoneally administered to rats 3 times with 24 h intervals, and the animals were sacrificed by bleeding at 24 h after the last administration. The hepatic regucalcin mRNA levels were markedly reduced by phenobarbital administration. This decrease was about 50% of control level with the 12 mg/100 g dose. Moreover, the hepatic regucalcin concentration was significantly decreased by the administration of phenobarbital (12 mg/100 g), although the serum regucalcin concentration was not altered appreciably. Meanwhile, serum transaminases (GOT and GPT) activities were not increased by the administration of phenobarbital (4 and 12 mg/100 g). The present study demonstrates that the expression of hepatic regucalcin mRNA is decreased by phenobarbital administration in rats, suggesting that regucalcin does not have a role in drug metabolism related to phenobarbital.
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  • 49
    ISSN: 1573-4919
    Keywords: vasoactive intestinal peptide ; ulcerative colitis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The availability of colon provides a ready source of human neurons. Among the products of nerve cell bodies, vasoactive intestinal peptide is a neuropeptide that serves as a marker of non-adrenergic, non-cholinergic inhibitory nerves in colon. These nerves have been proposed to be involved in regulation of immune function, secretion, and smooth muscle function. In previous work, we identified decreased tissue levels of vasoactive intestinal peptide in a disorder of chronic colonic mucosal inflammation, ulcerative colitis. We hypothesized that diminished gene expression of vasoactive intestinal peptide could result in decreased tissue levels of this neuropeptide. Sigmoid colon was obtained at surgery from controls (n=6) and patients with ulcerative colitis (n=6). Vasoactive intestinal peptide mRNA was quantified by Northern blot hybridization and tissue levels of vasoactive intestinal peptide were determined by radioimmunoassay. Tissue vasoactive intestinal peptide was decreased only in the mucosalsubmucosal layer of ulcerative colitis (p=.02). There was a single 1.7 kbase vasoactive intestinal peptide transcript identified in both control colon and ulcerative colitis. Normalized vasoactive intestinal peptide mRNA levels were increased by 260% in ulcerative colitis compared to controls (p〈.01). These observations suggest that decreased vasoactive intestinal peptide gene expression or abnormal post-transcriptional processing are not primary defects in this disorder of chronic inflammation. The findings support the alternative hypothesis that axonal degeneration in ulcerative colitis could result in increased expression of neuronal vasoactive intestinal peptide mRNA.
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  • 50
    ISSN: 1573-4919
    Keywords: pressure overload ; myocardium ; gene expression ; fibroblast ; extracellular matrix ; ventricular hypertrophy ; growth factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Transforming Growth Factor-beta1 (TGF-β1) is expressed in the heart by muscle and non-muscle cardiac cells.In vitro, cardiac myocytes and non-muscle cells including cardiac fibroblasts and endothelial cells respond to regulatory effects of TGF-β1. Expression of TGF-β1 in the heart is subject to regulation by hemodynamic stimuli. Increased expression of mRNA transcripts for TGF-β1 has been reported in several models of cardiac hypertrophy. The objective of this study was to determine the effect of TGF-β1 in the myocardium. TGF-β1 was injected intravenously. Expression of mRNA transcripts for functional and structural proteins was determined by Northern hybridization analysis. DNA synthesis was determined by measurement of3H-thymidine incorporation into ventricular DNA. The results showed differential regulation of mRNAs for myocyte- and non-myocyte-specific proteins in the heart of TGF-β1 treated rats. Moderate but statistically significant decrease in DNA synthesis was observed in the heart of TGF-β1 treated rats (37.5%, P〈0.025). Together, these data point to a physiological role for TGF-β1 in the heart. They further suggest that similar to its diversein vitro cell-specific regulatory effects, TGF-β1 may have multicellular targets in the heart. Effect of TGF-β1 alone or combined with those of other cytokines/hormones that come into play, as the result of its administration, may be responsible for altered gene expression and DNA synthesis in the myocardium. We propose that in experimental models of myocardial hypertrophy which are associated with increased expression of TGF-β1 in the heart, the contribution of regulatory effects of this growth factor to the manifestations of ventricular hypertrophy could be significant.
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  • 51
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; insulin ; calcium ; gene expression ; rat liver
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of refeeding on the expression of Ca2+-binding protein regucalcin mRNA in the liver of fasted rats was investigated. When rats were fasted overnight, the hepatic regucalcin mRNA level was reduced about 70% of that in feeding rats. Refeeding produced a remarkable elevation of hepatic regucalcin mRNA level (about 150–170% of fasted rats). Liver regucalcin concentration was appreciably increased by refeeding, although it was not altered by fasting. The oral administration of glucose (2 g/kg body weight) to fasted rats caused a significant increase in hepatic regucalcin mRNA level. Moreover, hepatic regucalcin mRNA level was clearly elevated by a single subcutaneous administration of insulin (10 and 100 U/kg) to fasted rats. The hormonal effect was not further enhanced by the simultaneous administration of calcium chloride (250 mg Ca/kg) to fasted rats, although calcium administration stimulated regucalcin mRNA expression in the liver. The present study suggests that the expression of hepatic regucalcin mRNA stimulated by refeeding is significantly involved in the action of insulin and/or calcium as stimulating factors.
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  • 52
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    Molecular and cellular biochemistry 143 (1995), S. 67-71 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; gene distribution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The existence and expression of gene encoding the Ca2+-binding protein regucalcin in various species and tissues were investigated with Southern and Northern hybridization analyses using regucalcin cDNA (0.9 kb of open reading frame). Genomic Southern hybridization analysis demonstrated that regucalcin gene was widely conserved among higher animals including human, monkey, rat, mouse, dog, bovine, rabbit and chicken. The gene was not found in yeast. The Northern blot analysis of poly (A)+RNAs extracted from the liver of various species showed that regucalcin mRNA was predominantly expressed in rat and mouse, although the expression was also seen in human, bovine and chicken. Furthermore, the enzyme-linked immunoadsorbent assay (ELISA) with rabbit-anti-regucalcin IgG indicated that hepatic regucalcin concentration was most pronounced in rat as compared with that of guinea pig, mouse and chicken. These observations show that the gene expression of regucalcin and its protein synthesis is unique in the liver of rats, suggesting the existence of a specific mechanism in demonstrating regucalcin synthesis from gene.
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  • 53
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    Molecular and cellular biochemistry 143 (1995), S. 137-141 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; estrogen ; gene expression ; rat liver
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of nuclear receptor-related hormones on the expression of hepatic calcium-binding protein regucalcin mRNA in rats was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin cDNA (0.9 kb of open-reading frame). A single subcutaneons administration of 17β-estradiol (0.5, 1.0 and 2.0 mg/kg body weight) in rats induced a remarkable increase of regucalcin mRNA in liver; the level was about 200% of control at 24 h after the administration of 2.0 mg/kg. The increase showed about 350% even at 6 h after the administration. Meanwhile, hepatic regucalcin mRNA level was not appreciably altered by a single subcutaneous administration of thyroxine (T4) (20, 40 and 80 mg/kg) or hydrocortisone (10 and 30 mg/kg) in rats. The present study demonstrates that the expression of hepatic regucalcin mRNA is stimulated by estrogen action in the liver nuclei of rats.
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  • 54
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    Molecular and cellular biochemistry 144 (1995), S. 105-108 
    ISSN: 1573-4919
    Keywords: fatty acid synthase ; gene expression ; and thyroid hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of triiodothyronine (T3) on regulation of fatty acid synthase in chicken liver was investigated. In hypothyroid animals, enzyme activity was about one half of that in euthyroid animals. T3 treatment increased the enzyme activity in hypothyroid animals. There is little difference in both the mRNA concentration and the transcription rate between euthyroid and hypothyroid animals. T3 treatment markedly decreased both the mRNA concentration and the transcription rate in euthyroid and hypothyroid animals. These results suggested that T3 maintained the normal level of enzyme expression primarily by stimulating the post-transcriptional step, while the transcription of the gene was inhibited by hyperthyroidism.
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  • 55
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    Molecular and cellular biochemistry 155 (1996), S. 85-90 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; rat hepatoma ; Morris hepatoma cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Whether the gene expression of hepatic Ca2+-binding protein regucalcin is altered in hepatomas was investigated. The change in regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb). Rat hepatoma was induced by continuous feeding of basal diet containing 0.06% 3′-methyl-4-dimethylaminoazobenzene (3′-Me-DAB). After 35 weeks feeding, rats were sacrificed, and the non-tumorous and tumorous tissues of the livers were removed. In individual rats, the regucalcin mRNA levels in the tumorous tissues were generally decreased in comparison with that of the non-tumorous tissues of the chemical-fed rats, although the chemical administration might decrease the mRNA expression in normal rat liver, suggesting that the chemical administration causes a suppresive effect on the mRNA expression. When the genomic DNA extracted from the liver tumorous tissues was digested with restriction enzymes (EcoRI, BamHI and HindIII) and analyzed by Southern blotting, no rear-ranged band was found in the regucalcin gene from the hepatoma. Interestingly, in the transplantable Morris hepatoma cells, the regucalcin mRNA was markedly expressed, while the albumin mRNA was expressed only slightly. The present study demonstrates that regucalcin mRNA is clearly expressed in the transformed cells (Morris hepatoma cells).
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  • 56
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    Molecular and cellular biochemistry 155 (1996), S. 105-111 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; aldosterone ; estrogen ; dexamethasone ; gene expression ; rat kidney cortex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of various steroid hormones on the expression of calcium-binding protein regucalcin mRNA in the kidney cortex of rats was investigated. The change of 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 the kidney cortex but not the medulla. Rats received a single subcutaneous administration of steroid; the animals were sacrificed 60 min after the treatment of aldosterone (2.5, 5.0 and 10 μg/100 g body weight) or 6 h after the treatment of estrogen (17β-estradiol; 0.05, 0.1 and 0.2 mg/100 g), hydrocortisone (0.5, 1.0 and 3.0 mg/100 g) and dexamethasone (50, 100 and 150 μg/100 g). Regucalcin mRNA levels in the kidney cortex were clearly diminished by the administration of aldosterone or estrogen, while hydrocortisone administration had no effect. The administration of dexamethasone (100 μg/100 g) caused a remarkable increase of regucalcin mRNA levels in the kidney cortex. The dexamethasone-induced increase in regucalcin mRNA levels was completely blocked by the simultaneous administration of cycloheximide (150 μg/100 g), although the drug administration had no effect on the mRNA levels in control rats. Meanwhile, the dexamethasone administration did not cause an appreciable alteration of calcium content in the kidney cortex. The present study demonstrates that, of the various steroid hormones used, dexamethasone uniquely has a stimulatory effect on regucalcin mRNA expression in the kidney cortex of rats. The steroid effect may be mediated through a newly synthesized protein.
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  • 57
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    Molecular and cellular biochemistry 160-161 (1996), S. 307-313 
    ISSN: 1573-4919
    Keywords: myocardium ; hypertension ; gene expression ; estrogens ; cardiac hypertrophy ; signal transduction ; genetic program
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Gender specific differences in cardiovascular disease are largely mediated by sex hormones. The use of estrogens significantly reduces the overall incidence of heart disease in postmenopausal women. Beneficial effects of estrogens on plasma lipoprotein levels are clearly established. However, these do not explain the magnitude of risk reduction seen in clinical studies. Thus, additional and currently unknown functions of estrogens must be operative. Elucidation of the exact estrogen action in the heart will have important implications in the treatment of cardiovascular disease. It will probably enhance the therapeutic repertoire in treating heart disease, the most common cause of death in industrialized countries. We will review the current understanding of the function of estrogens in the heart and discuss potential strategies on how to apply these data to clinical practice.
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  • 58
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    Molecular and cellular biochemistry 162 (1996), S. 139-144 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; saline ingestion ; hypertensive rats ; kidney cortex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The effect of adrenalectomy (ADX) or saline ingestion, which is a hypertensive factor, on the expression of calcium-binding protein regucalcin mRNA in the kidney cortex of rats was investigated. The change of 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 the kidney cortex but not the medulla. Rats were adrenalectomized, and 48 h later they were sacrificed. ADX caused a reduction of regucalcin mRNA levels in the kidney cortex, suggesting that adrenal glands participate in the regulation of the mRNA expression. This reduction was not restored by the subcutaneous administration of dexamethasone with an effective dose (1 mg/kg body weight), which can stimulate kidney regucalcin mRNA expression. Regucalcin mRNA levels in the kidney cortex of rats were markedly suppressed by the ingestion of saline for 7 days. The ADX-induced decrease of renal cortex regucalcin mRNA levels was not appreciably restored by saline ingestion. Moreover, regucalcin mRNA levels in the kidney cortex of spontaneous hypertensive rats (SHR) were clearly decreased as compared with that of control (Wistar-Kyoto) rats. Meanwhile, calcium content in the kidney cortex was not significantly decreased by ADX or saline ingestion. The present study suggests that the expression of regucalcin mRNA in the kidney cortex of rats is suppressed by saline administration.
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  • 59
    ISSN: 1573-4919
    Keywords: calreticulin ; gene expression ; steroid receptor
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    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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  • 60
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    Molecular and cellular biochemistry 148 (1995), S. 45-57 
    ISSN: 1573-4919
    Keywords: manganese ; superoxide dismutase ; gene expression ; hyperoxide lung injury ; nuclear factor kappa B
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract TNFα and IL-1 each can activate NF-κB and induce gene expression of manganese superoxide dismutase (MnSOD), a mitochondrial matrix enzyme which can provide critical protection against hyperoxic lung injury. The regulation of MnSOD gene expression is not well understood. Since redox status can modulate NF-κB and potential κB site(s) exist in the MnSOD promoter, the effect of thiols (including NAC, DTT and 2-ME) on TNFα and IL-1 induced activation of NF-κB and MnSOD gene expression was investigated. Activation of NF-kB and increased MnSOD expression were potentiated by thiol reducing agents. In contrast, thiol oxidizing or alkylating agents inhibited both NF-κB activation and elevated MnSOD expression in response to TNFα or IL-1. Since protease inhibitors TPCK and TLCK can inhibit NF-κB activation, we also investigated the effect of these compounds on MnSOD expression and NF-κB activation. TPCK and TLCK each inhibited MnSOD gene expression and NF-κB activation. Since the MnSOD promoter also contains anAP-1 binding site, the effect of thiols and thiol modifying agents on AP-1 activation was investigated. Thiols had no consistent effect onAP-1 activation. Likewise, some of the thiol modifying compounds inhibited AP-1 activation by TNFα or IL-1, whereas others did not. Since diverse agents had similar effects on activation of NF-κB and MnSOD gene expression, we have demonstrated that activation of NF-κB and MnSOD gene expression are closely associated and that reduced sulfhydryl groups are required for cytokine mediation of both processes.
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  • 61
    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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  • 62
    ISSN: 1573-4919
    Keywords: microbodies ; diabetes mellitus ; steroid hormone receptor ; β-oxidation ; gene expression
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract To determine whether the increased fatty acid β-oxidation in the peroxisomes of diabetic rat liver is mediated by a common peroxisome proliferation mechanism, we measured the activation of long-chain (LC) and very long chain (VLC) fatty acids catalyzed by palmitoyl CoA ligase (PAL) and lignoceryl CoA ligase and oxidation of LC (palmitic acid) and VLC (lignoceric acid) fatty acids by isotopic methods. Immunoblot analysis of acyl-CoA oxidase (ACO), and Northern blot analysis of peroxisome proliferator-activated receptor (PPAR-α), ACO, and PAL were also performed. The PAL activity increased in peroxisomes and mitochondria from the liver of diabetic rats by 2.6-fold and 2.1-fold, respectively. The lignoceroyl-CoA ligase activity increased by 2.6-fold in diabetic peroxisomes. Palmitic acid oxidation increased in the diabetic peroxisomes and mitochondria by 2.5-fold and 2.7-fold, respectively, while lignoceric acid oxidation increased by 2.0-fold in the peroxisomes. Immunoreactive ACO protein increased by 2-fold in the diabetic group. The mRNA levels for PPAR-α, ACO and PAL increased 2.9-, 2.8- and 1.6-fold, respectively, in the diabetic group. These results suggest that the increased supply of fatty acids to liver in diabetic state stimulates the expression of PPAR-α and its target genes responsible for the metabolism of fatty acids.
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  • 63
    ISSN: 1573-4927
    Keywords: mutator ; transposable element ; alcohol dehydrogenase ; maize ; gene expression
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A secondary mutant, derived from an allele of maize alcohol dehydrogenase 1 (Adh1) carrying a Mutator transposable element (Mu1) in its first intron, was reported to exhibit a threefold decrease in ADH enzymatic activity and steady-state RNA levels compared to the original mutant. The original mutant,Adh1-S3034 (abbreviatedS3034), was previously characterized at the molecular level. The derivative, abbreviatedS3034b, has now been cloned; at the DNA sequence level the insertion and surroundingAdh1 sequences are indistinguishable fromS3034. Furthermore, in our lines there is no difference in relative ADH activities between products of the two putative alleles. A comparison of gene expression in heterozygotes obtained by crossing to different tester lines reveals a correlation between the measured decrease in levels of ADH polypeptide produced by the mutant allele and the background in which it is measured; this effect is distinct from any background-related variation in the expression of the progenitor allele. It does not appear to be attributable to alternative patterns of DNA modification. It appears to reflect a background-associated difference in the level of normalAdh1-RNA produced. Thus the previously reported distinction betweenS3034 andS3034b may be due to differences in the extent to which the mutant allele and a given genetic background interact to produce functionalAdh1-RNA.
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  • 64
    ISSN: 1573-4919
    Keywords: Rb and p53 genes ; gene expression ; colorectal cancers ; colon carcinoma cell lines ; cell cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We have compared the expression of the retinoblastoma (Rb) and p53 genes in normal human fibroblasts, colon carcinoma cell lines, matched pairs of colorectal tumor tissues and adjacent normal mucosa and in synchronized human diploid fibroblast cell line W138. The increased expression of Rb and p53 RNA was observed in a majority of colorectal cancers in comparison to adjacent normal mucosa and is accompanied by proportional increase in the expression of histone H3 gene. The Rb and p53 RNA levels varied significantly between the various colon carcinoma cell lines. However, we found that the expression of Rb and p53 RNA is regulated differently in cell cycle synchronized normal human fibroblasts. The Rb mRNA level did not change with the position in the cell cycle and did not differ significantly whether the cells were serum deprived or in 10% serum. But p53 mRNA expression follows the same pattern as histone H3 mRNA.
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  • 65
    ISSN: 1573-4919
    Keywords: mouse ; protein tyrosine phosphatase ; cDNA cloning ; nucleotide sequence ; gene expression
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The PTP-2 cDNA encoding an intracellular protein tyrosine phosphatase (PTPase-2) was isolated and sequenced from mouse testis and T-cell cDNA libraries. This PTP-2 cDNA was found to be homologous to human PTP-TC and rat PTP-S, and contained 1,551 nucleotides, including 1,146 nucleotides encoding 382 amino acids as well as 5′ (61 nucleotides) and 3′ (344 nucleotides) non-coding regions. Northern blot analysis indicated that PTP-2 mRNA of 1.9 Kb was most abundant in testis and kidney, although it was also present in spleen, muscle, liver, heart and brain.
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  • 66
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    Molecular and cellular biochemistry 135 (1994), S. 79-88 
    ISSN: 1573-4919
    Keywords: calcium ; nucleus ; calpain ; calmodulin ; cell division ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Calcium has long been known to play a role as a key cytoplasmic second messenger, but until relatively recently its possible involvement in nuclear signal transduction and the regulation of nuclear events has not been extensively studied. Evidence revealing the presence of transmembrane nuclear Ca2+ gradients and a variety of intranuclear Ca2+ binding proteins has fueled renewed interest in this key ion and its involvement in cell-cycle timing and division, gene expression, and protein activation. This review will offer an overview of the current state of knowledge and theory regarding calcium orchestration of nuclear functions and events and discuss possible future directions in this field of study.
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  • 67
    ISSN: 1573-4919
    Keywords: poly(ADP-ribosyl) transferase (human) ; autoregulation ; gene expression ; promoter structure ; cruciform structure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Human nuclear poly(ADP-ribosyl)transferase (ADPRT) modifies proteins with branched ADP-ribose-polymers. Various proteins, including ADPRT itself, serve as acceptors for polyADP-ribose. Target proteins include those controlling basic cellular processes such as DNA repair, differentiation and proliferation. Because of the outstanding features of this enzyme: automodification, several functional domains and central role in physiology of the cell, the molecular biology of ADPRT gained wide interest. The promoter structure contains several CCAAT/TATA boxes and SP1 sites. However, there is no CCAAT/TATA box in the neighbourhood of an SP1 site and, thus no obvious site for initiation of transcription. Within this region there are several noteworthy inverted repeats, which by internal basepairing could form two types of cruciform structures. Deletion analysis revealed that these cruciform structures have functional significance. Removal of one type increases the promoter activity, whereas removal of the other diminishes the promoter function. Overexpression of ADPRT from heterologous promoters (MMTV, SV40) leads to repression of the activity of the ADPRT promoter. Indeed, ADPRT was shown to bind specifically to one type of cruciform structure. This specific interaction indicates autorepression of the ADPRT gene: the enzyme ADPRT acts directly as a negative modulator of the activity of its own promoter.
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  • 68
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    Molecular and cellular biochemistry 155 (1996), S. 153-162 
    ISSN: 1573-4919
    Keywords: apolipoprotein B and E ; lipid ; gene expression ; rat ; mouse
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The aim of the present investigation was to study the regulation of apolipoprotein E by two dietary nutrients, saturated fat and cholesterol, known to raise plasma cholesterol levels. ApoE is a protein component of several classes of lipoproteins including VLDL and HDL, and dietary lipids may regulate VLDL and apoE-containing HDL particles through their effects on apoE gene. Male rats and mice were fed the following 4 diets: control diet (C); high cholesterol diet with 0.5% cholesterol (HC); high fat diet with 20% hydrogenated coconut oil (HF); and high fat plus high cholesterol diet with 0.5% cholesterol and 20% fat (HF/C). Plasma cholesterol levels remained unchanged on HC diet, but in mice VLDL-cholesterol increased by 31%. HF diet increased VLDL and LDL by 15–17% in rats, and 21% in mice. A combination of fat and cholesterol diet showed pronounced effects on plasma lipoprotein concentrations, raising apoB-containing particles by 21% and 44% in mice and rats, respectively. Plasma apoE levels increased significantly on all diets. The mechanism of regulation of increased plasma apoB and apoE levels was examined. Quantification of hepatic apoB mRNA showed a lack of correlation between plasma apoB and hepatic apoB mRNA levels, suggesting that posttranscriptional regulation increased plasma apoB-containing lipoproteins in animals fed saturated fat diets. Hepatic apoE mRNA levels increased significantly in animals fed cholesterol-rich diets. However, despite increased plasma apoE levels on diet containing only saturated fat, hepatic apoE mRNA did not change. Synthesis of apoE on the liver polysomes increased selectively on cholesterol-rich diets. These results suggest that cholesterol-rich diets altered apoE, in part, by transcriptional mechanism, and saturated fat-rich diets increased plasma apoE levels by posttranscriptional mechanism, possibly decreased receptor-mediated uptake of apoE-containing particles. The regulation of LDL receptor was also studied since plasma apoB and E levels may be altered by LDL receptor-mediated uptake by the hepatocytes. As expected, high cholesterol diet decreased LDL receptor mRNA by 30–40%. However, the LDL receptor protein on liver membranes did not change on any of the test diets in both animal species. Hepatic cholesterol content increased several fold selectively on high cholesterol diets. These findings suggest that: 1) both transcriptional and posttranscriptional mechanisms are important in regulating plasma apoB and E containing lipoproteins; 2) dietary cholesterol regulates apoE gene by a transcriptional mechanism anddietary saturated fat by posttranscriptional mechanism; and 3) changes in the hepatic apoE and LDL receptor mRNA are associated with the changes in intracellular cholesterol concentrations.
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  • 69
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    Molecular and cellular biochemistry 146 (1995), S. 71-77 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; 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 was investigated. The change of regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb of open-reading frame). Regucalcin mRNA was expressed in the kidney cortex, and this expression was clearly increased by a single intraperitoneal administration of calcium chloride solution (5–15 mg Ca/100 g body weight) in rats; this increase was remarkable at 60–120 min after the administration. Thyroparathyroidectomy (TPTX) caused a slight decrease of regucalcin mRNA levels in the kidney cortex. However, the administration of calcium (10 mg/100 g) in TPTX rats produced a clear increase of regucalcin mRNA levels in the kidney cortex. The subcutaneous administration of calcitonin (10–100 MRC mU/100 g) or parathyroid hormone [1–34] (1–10 U/100 g) in TPTX rats which received calcium (10 mg/100 g) administration did not cause an appreciable alteration of regucalcin mRNA levels in the kidney cortex, suggesting that the mRNA expression is not stimulated by calcium-regulating hormones. The administration of trifluoperazine (TFP; 5 mg/100 g), an inhibitor of Ca2+/calmodulin action, completely blocked the expression of regucalcin mRNA stimulated by calcium administration. Now, calcium content in the kidney cortex was significantly elevated by a single intraperitpneal administration of calcium (10 mg/100 g) in rats. The present study clearly demonstrates that the expression of regucalcin mRNA in the kidney cortex is stimulated by calcium administration in rats. This expression may be mediated through Ca2+/calmodulin action in the kidney cortex.
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  • 70
    ISSN: 1573-4919
    Keywords: fatty acid transport protein ; gene expression ; subtractive hybridization ; oxidative stress ; ischemia/reperfusion ; ischemic preconditioning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract In this study, ischemia and oxidative stress-inducible gene expression in heart was examined by subtractive hybridization technique. Total RNA was isolated from ventricular muscle fragments of normal and oxidative stress-induced hearts. Poly (A)+ RNA was purified followed by the construction of a plasmid cDNA library. This was followed by the subtractive screening of oxidative stress-induced cDNA library. The positive colonies were amplified and the plasmid isolated. An aliquot was subjected to restriction cutting with Bam H1 and EcoRl; the fragments corresponding to cDNA insert were separated by electrophoresis, radiolabeled by random-primed DNA synthesis, and used as probes in standard Northern blotting experiments. An aliquot containing the plasmid from the confirmed positives was then subjected to bidirectional partial DNA sequencing (using M13 and T7/T3α primers) by the chain-extension/chain termination method. These sequences were subjected to a computerized search for homologies against all sequences in the updated worldwide Gen Bank and EMBL sequence databases followed by restriction mapping and reading frame identification. Out of 24 putative positive colonies screened, one clone was matched with 〉 97% homology with FAT gene that has been implicated in binding or transport of long chain fatty acids. cDNA probe synthesized from this clone identified two major transcripts of 4.8 and 2.9 kb. Additional experiments were then performed where isolated perfused rat hearts were subjected to the following treatments: (1) 5 min ischemia; (2) 10 min ischemia; (3) 20 min ischemia; (4) 5 min ischemia followed by 10 min reperfusion (ischemic preconditioning); and (5) 5 min ischemia followed by 10 min reperfusion, repeated four times (4 × preconditioning). RNAs were extracted from these hearts and hybridized with the FAT cDNA probe. The results indicated that FAT gene was induced by oxidative stress, ischemic preconditioning, but not by ischemia. (Mol Cell Biochem 160/161: 241–247, 1996)
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  • 71
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    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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  • 72
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    Molecular and cellular biochemistry 163-164 (1996), S. 231-237 
    ISSN: 1573-4919
    Keywords: extracellular matrix ; angiotensin II ; fibrillar collagen ; cardiac fibrosis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Recent studies suggest that angiotensin II (angiotensin) may be involved in the regulation of metabolism of the cardiac extracellular matrix (ECM). Two major components of ECM are collagen types I and III which play an important role in maintaining the structure and function of the heart. Although the cellular metabolism of collagen is very complex (especially at the posttranslational level), we chose to address events that occur relatively early in the synthesis of cardiac collagen molecules. To gain an understanding of the role of angiotensin in the regulation of cardiac collagen gene expression, we studied the effect of three different doses of angiotensin (12, 24, and 48 μg/kg/h) on adult heart and cultured neonatal cardiac fibroblasts. The steady-state mRNA abundance of collagen types I and III was monitored using Northern blot analysis in both left and right ventricular samples at day 3 of angiotensin infusion and in cultured cardiac fibroblasts stimulated with angiotensin. In all mRNA abundance studies, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) signal was used to normalize the data for possible differences in loading and/or transfer of total RNA. Both collagen types I/GAPDH and III/GAPDH mRNA signal ratios were increased significantly in left ventricle in all dose regimens used for angiotensin infusion. Only the collagen type I/GAPDH mRNA signal ratio was increased in right ventricle with angiotensin infusion. Angiotensin (10−7-10−5 M) had no effect on the steady-state mRNA abundance of collagen genes in cultured neonatal cardiac fibroblasts after 24 h treatment in serum-free conditions. Our results confirm that infusion of angiotensin may upregulate steady-state collagen gene mRNA abundance in the heart. Angiotensin had no observable effect on collagen mRNA abundance in neonatal fibroblast culture. An explanation for the current results may be that angiotensin causes the release of undefined factors from cardiac myocytes, and that these secondary factors may be involved in either the activation of collagen gene transcription or in alteration of stability of collagen mRNA transcripts via a paracrine mechanism. Although our results indicate hemodynamic loading may potentiate the action of angiotensin, this scenario is unlikely as collagen type I gene expression was increased in the normotensive right ventricle.
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  • 73
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    Molecular and cellular biochemistry 167 (1997), S. 169-177 
    ISSN: 1573-4919
    Keywords: tamoxifen ; interferon ; gene expression ; breast cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The molecular basis for the enhanced growth inhibition of MCF-7 human breast cancer xenografts by a combination of human interferon-β (IFN-β) and tamoxifen was investigated. Treatment of MCF-7, MDA-MB-231, and BT-20 cells with the combination of IFN-β and tamoxifen resulted in enhanced antiproliferative effects in vitro. Treatment with the combination of IFN-β and tamoxifen enhanced the expression of several IFN-β-inducible genes in human breast carcinoma cell lines relative to levels induced by IFN-β alone. Tamoxifen alone did not induce transcription of IFN-stimulated genes (ISGs). Augmentation of ISG expression by the combination of IFN-β and tamoxifen was noted in breast tumor cell lines irrespective of their functional estrogen receptor (ER) status or their dependence on estradiol for growth, suggesting that upregulation of ISGs was independent of ER status. Enhancement of IFN-stimulated gene expression by tamoxifen occurred at the transcripti onal level. Expression of transfected reporter genes under the control of IFN-α/β regulated promoters was also enhanced in IFN-β and tamoxifen-treated cells. Similarly, transcriptional induction of chimeric reporter plasmids driven by an IFN-γ inducible promoter (GAS; IFN-γ activated site) was also enhanced by the combination of IFN-γ and tamoxifen. In tamoxifen treated cells, IFN-β and IFN-γ readily activated transcription factors ISGF-3 and GAF, respectively. Therefore, augmentation of ISG expression by tamoxifen is an early event in the antitumoral activity of this drug combination. (Mol Cell Biochem 167: 169-177, 1997)
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  • 74
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    Molecular and cellular biochemistry 172 (1997), S. 47-57 
    ISSN: 1573-4919
    Keywords: smooth muscle ; gene transfer ; DNA ; RNA ; ribozyme ; liposome ; lipoxygenase ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Chemically synthesized hammerhead-type ribozymes targeted against the porcine leukocyte-type 12-lipoxygenase (LO) have been developed and studied. One chimeric ribozyme consists of DNA in the non-enzymatic portions, and RNA in the enzymatic core as well as two phosphorothioate internucleotide linkages at 3′ terminus. The second ribozyme consists of ribonucleotide sequences generated by in vitro transcription. In this chapter we describe methodologies to first analyze the ribozyme catalytic activity in vitro by studying cleavage of target RNA in vitro. The subsequent sections will describe how to target the catalytic ribozyme and deliver it to porcine vascular smooth muscle cells (PVSMC) by a liposome-mediated method. Finally ways to evaluate its activity to inhibit expression of the 12-LO mRNA will be presented. These results demonstrate the feasibility of using ribozymes as novel candidates for therapeutic agents to block specific gene expression in vascular cells.
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  • 75
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    Keywords: heart ; DNA ; library ; gene expression
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    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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  • 76
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    Molecular and cellular biochemistry 173 (1997), S. 59-69 
    ISSN: 1573-4919
    Keywords: hydrogen peroxide ; oxidative stress ; gene expression ; lens epithelial cells ; N-acetylcysteine ; pyrrolidine dithiocarbamate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The involvement of H2O2 in cataract development has been established inboth human patients and animal models. At the molecular level H2O2 has beenobserved to cause damage to DNA, protein and lipid. To explore the oxidativestress response of the lens system at the gene expression level, we haveexamined the effects of H2O2 on the mRNA change of the proto-oncogenes,c-jun, c-fos and c-myc in a rabbit lens cell line, N/N1003A. H2O2 treatmentof the rabbit lens epithelial cells for 60 min induces quick up-regulationof both c-jun and c-fos mRNAs. The maximal induction is 38 fold for c-jun at150 µM and 72 fold for c-fos at 250 µM H2O2. Treatment ofN/N1003A cells with 50-250 µM H2O2 for 60 min leads to a 2-5 foldincrease of the c-myc mRNA level. H2O2 also induces an up-regulation intransactivity of the activating protein-1 (AP-1) as shown with a reportergene driven by a prolactin gene promoter with 4 copies of AP-1 binding sitesinserted in the upstream of the promoter. Maximal induction occurs with 150µM H2O2. In the same system, the antioxidants, N-acetyl-cysteine (NAC)and pyrrolidine dithiocarbamate (PDTC) at concentrations shown toup-regulate the mRNAs of both c-jun and c-fos, also enhance thetransactivity of AP-1. NAC and PDTC have different effects in modulating theinduction of AP-1 activity by H2O2 and TPA. These results reveal thatoxidative stress regulates expression of various regulatory genes in lenssystems, which likely affects cell proliferation, differentiation andviability and thus affect normal lens functions.
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  • 77
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    Keywords: regucalcin ; calcium-binding protein ; cDNA cloning ; gene expression ; mouse liver
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    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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  • 78
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    Molecular and cellular biochemistry 176 (1997), S. 273-279 
    ISSN: 1573-4919
    Keywords: cardiac hypertrophy ; myosin heavy chain ; gene expression ; adrenergic system
    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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  • 79
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    Keywords: regucalcin ; Ca2+-binding protein ; protein kinase C ; Ca2+signaling ; gene expression ; H4-II-E hepatoma 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 rat hepatoma cells (H4-II-E) 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 H4-II-E hepatoma cells. This expression was clearly stimulated in the presence of serum (10% fetal bovine serum). Bay K 8644 (2. 5 × 10-6 M), a Ca2+ channel agonist, significantly stimulated regucalcin mRNA expression in the absence or presence of 10% serum. Dibutyryl cyclic AMP (10-3 M) did not have a stimulatory effect on the regucalcin mRNA expression. The presence of phorbol 12-myristate 13-acetate (PMA; 10-6 M) or estrogen (10-8 M) caused a significant increase in regucalcin mRNA levels in the hepatoma cells cultured in serum-free medium, while insulin (5 × 10-9 M) or dexamethasone (10-6 M) had no effect. Bay K 8644-stimulated regucalcin mRNA expression in the hepatoma cells was completely blocked in the presence of trifluoperazine (10-5 M), an antagonist of calmodulin, or staurosporine (10-7 M), an inhibitor of protein kinase C. The stimulatory effect of PMA was clearly inhibited in the presence of stauroporine. The present study demonstrates that regucalcin mRNA is expressed in the transformed H4-II-E hepatoma cells, and that the expression is stimulated through Ca2+-dependent signaling factors.
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  • 80
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    Molecular and cellular biochemistry 199 (1999), S. 189-200 
    ISSN: 1573-4919
    Keywords: lung ; cancer ; urokinase ; receptor ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The urokinase-type plasminogen activator (uPA) interacts with its receptor (uPAR) to promote proteolysis as well as cell proliferation and migration. These functions contribute to the pathogenesis of neoplastic growth and invasiveness. Expression of uPAR in tumor extracts also inversely correlates with prognosis in many forms of cancer. In this study, we sought to determine if differences in uPAR expression were distinguishable between cultured human lung carcinoma and malignant mesothelioma subtypes. We also sought to determine if, as in malignant mesothelioma cells, uPAR expression is regulated at the posttranscriptional level in cultured malignant lung carcinoma cells. Using 125I-uPA binding and ligand blotting techniques, uPAR was expressed by phenotypically diverse lung carcinoma cell lines, including the H460, H157 and H1395 non-small cell lines and the H146 small cell lung carcinoma line. Increased uPAR expression was also detected in spindle-shaped (M33K) and epithelioid (M9K and MS-1) malignant mesothelioma cells. Selected mediators, including TGF-β, TNF-α, LPS and PMA, uniformly enhanced uPAR expression in each of the tumor cell lines. Steady state uPAR mRNA expression was determined by RNase protection assay and correlated directly with the changes in cell surface uPAR expression. By gel mobility shift and UV-cross linking assays, a uPAR mRNA binding protein (uPAR mRNABp) implicated in the posttranscriptional control of message stability, was identified in each of the cell lines. Expression of uPAR and its message in cultured lung carcinoma and malignant mesothelioma cells is similarly influenced by effectors present in the tumor microenvironment. Regulation of the uPAR message occurs at the posttranscriptional level in cultured small and non-small cell lung carcinoma cells as well as spindle-shaped and fibrous malignant mesothelioma cell lines. Posttranscriptional regulation of uPAR in all these cells involves the interaction of the uPAR mRNABp with uPAR mRNA, which promotes uPAR mRNA destabilization.
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  • 81
    ISSN: 1573-4927
    Keywords: glycophorins ; gorilla ; evolution ; gene family ; gene expression
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    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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    Biochemical genetics 28 (1990), S. 337-346 
    ISSN: 1573-4927
    Keywords: glutamate dehydrogenase ; Drosophila melanogaster ; gene expression ; evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We have isolated theDrosophila melanogaster locus coding forl-glutamate dehydrogenase (EC 1.4.1.3) by virtue of its similarity to the corresponding human gene. There is only one copy of this gene in the fruit fly genome, located on the right arm of chromosome 3 (95D1-4). The transcript includes at least one large intron and matures to a ∼2.4-kb-long polyadenylated RNA whose expression is under developmental control.
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  • 83
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    Keywords: glucocorticoid receptor ; MMTV ; transcription factors ; gene expression
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The long terminal repeat (LTR) of the mouse mammary tumor virus was used as a template to examine the dual binding parameters of the glucocorticoid-receptor (GR) and a repressor protein termed Inhibitory Factor 1 (IF1). The roceptor binds specifically to the glucocorticoid response element and precludes the binding of IF1 to its juxtaposed binding site within the LTR. When the two DNA targets are separated by the insertion of an additional 52 base pairs, coincident binding of both proteins is observed. Gel retention assays reveal three distinct nucleoprotein complexes. The first complex consists of the receptor and the LTR, the second is comprised of IF1 and DNA and the third is a multiprotein-DNA complex consisting of the GR, IF1 and DNA, migrating at a higher molecular weight position. The inhibition of IF1 binding by the presence of prebound GR leads to the repression of transcription of juxtaposed genes. The GR may act to block access of a sequence, used by the cell to titrate repressor proteins and facilitate the onset of gene expression. (Mol Cell Biochem122: 25–37, 1993)
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    Molecular and cellular biochemistry 122 (1993), S. 147-158 
    ISSN: 1573-4919
    Keywords: PSG transcripts ; gene expression ; PCR ; T lymphocytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The presence of PSG in blood cells has been demonstrated by immunohistochemical staining. However, the origin of these proteins is not known. This report examines the expression of the PSG genes in different types of freshly isolated blood cells. RNA isolated from bone marrow and peripheral blood cells of healthy individuals was analyzed for PSG transcripts by reverse transcriptase-polymerase chain reaction using synthetic oligonucleotide primers specific for the PSG genes. The level of expression of the PSG genes in different types of cells exhibited significant individual variation. Trace amounts of PSG transcripts could be detected in polymorphonuclear cells (PMN), monocytes and B lymphocytes while T lymphocytes always contained the highest level of transcript. The expression of PSG genes in the blood cells apparently was not affected by the method of isolation nor by overnight culturing of these cells except in the case when lymphocytes were separated by rosetting with sheep red blood cells. All reported PSG transcripts were detected in blood cells. Both type I and type II transcripts of the PSG genes were detected in blood cells with the exception of type II transcript of PSG5 and PSG11 which were only found in the placenta. Tissue specificity in the expression or alternative splicing of some of the PSG family members was implicated.
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  • 85
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    Molecular and cellular biochemistry 133-134 (1994), S. 235-243 
    ISSN: 1573-4919
    Keywords: creatine kinase ; mitochondria ; metabolism ; creatine phosphate shuttle ; gene expression ; muscle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Mitochondrial Creatine Kinase (MtCK) is responsible for the transfer of high energy phosphate from mitochondria to the cytosolic carrier, creatine, and exists in mammals as two isoenzymes encoded by separate genes. In rats and humans, sarcomere-specific MtCK (sMtCK) is expressed only in skeletal and heart muscle, and has 87% nucleotide identity across the 1257 bp coding region. The ubiquitous isoenzyme of MtCK (uMtCK) is expressed in many tissues with highest levels in brain, gut, and kidney, and has 92% nucleotide identity between the 1254 bp coding regions of rat and human. Both genes are highly regulated developmentally in a tissue-specific manner. There is virtually no expression of sMtCK mRNA prior to birth. Unlike cytosolic muscle CK (MCK) and brain CK (BCK), there is no developmental isoenzyme switch between the MtCKs. Cell culture models representing the tissue-specific expression of either sMtCK or uMtCK are available, but there are no adequate developmental models to examine their regulation. Several animal models are available to examine the coordinate regulation of the CK gene family and include 1) Cardiac Stress by coarctation (sMtCK, BCK, and MCK), 2) Uterus and placenta during pregnancy (uMtCK and BCK), and 3) Diabetes and mitochondrial myopathy (sMtCK, BCK, and MCK). We report the details of these findings, and discuss the coordinate regulation of the genes necessary for high-energy transduction.
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    Molecular and cellular biochemistry 135 (1994), S. 89-98 
    ISSN: 1573-4919
    Keywords: calcium ; cell death ; nuclei ; apoptosis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The last decade has seen the rapid development of research investigating the molecular mechanisms whereby hormones, peptide growth factors and cytokines regulate cell metabolism, differentiation and proliferation. One general signalling mechanism used to transfer the information delivered by agonists into appropriate intracellular compartments involves the rapid Ca2+ redistribution throughout the cell, which results in transient elevations of the cytosolic free Ca2+ concentration. Ca2+ signals are required for a number of cellular processes including the activation of nuclear processes such as gene transcription and cell cycle events. The latter require that appropriate Ca2+ signals elicited in response to agonists be transduced across the nuclear envelope. It has generally been assumed that small molecules, metabolites and ions could freely diffuse across the nuclear envelope. Nevertheless several findings during the past few years have suggested that nuclear pore permeability can be regulated and that ion transport systems and ion-selective channels may exist on the nuclear membranes and regulate intranuclear processes. Intranuclear Ca2+ fluctuations can affect chromatin organization, induce gene expression and also activate cleavage of nuclear DNA by nucleases during programmed cell death or apoptosis. The possible mechanisms involved in nuclear Ca2+ transport and the control of nuclear Ca2+-dependent enzymes in apoptosis is discussed below.
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    Molecular and cellular biochemistry 103 (1991), S. 149-154 
    ISSN: 1573-4919
    Keywords: EGF-receptor gene ; gene expression ; cycloheximide ; dexamethasone and A431 carcinoma cell line
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Human A431 carcinoma cell line is known to have 30 fold amplified epidermal growth factor receptor (EGF-R) gene. We have studied the effect of steroid hormone dexamethasone (DEX) and protein synthesis inhibitor cycloheximide (CHX) on the expression of EGF-R gene in this cell line. DEX treatment and protein synthesis inhibition by CHX treatment cause a rapid 3 to 4 fold increase in the level of EGF-R mRNA and combined treatment of the above two agents have less than additive effect. It appears that mRNA for EGF-R accumulate within the cell during protein synthesis inhibition and upon removal of CHX, gets translated into EGF-R specific protein as judged by immuno-dot assay. We did not observe the phenomenon of ‘super induction’ nor much of an additive effect under condition of combined DEX and CHX treatment.
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    Molecular and cellular biochemistry 104 (1991), S. 35-43 
    ISSN: 1573-4919
    Keywords: α1-adrenergic receptors ; β-adrenergic receptors ; cardiac muscle ; cell culture ; gene expression ; protein kinase C
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The molecular mechanisms of cardiac myocyte growth are relevant to important problems in cardiovascular disease. A cell culture model has been developed to explore the role of adrenergic hormones in cardiac myocyte growth and gene expression. Activation of a cardiac myocyte α1-adrenergic receptor by catecholamines induces hypertrophic growth of neonatal rat cardiac myocytes and initiates selective increases in contractile protein gene transcription. These effects on growth and gene expression do not depend on contractile activity. The cardiac myocytes contain at least two subtypes of α1-adrenergic receptors and at least three isoforms of protein kinase C (PKC). A distinct α1 receptor subtype may mediate hypertrophy and gene transcription. Different isoforms of PKC are translocated to different intracellular sites on activation, and there is evidence that the β-PKC isoform may be an element in the signal transduction pathway from an α1 receptor at the surface to the cardiac myocyte nucleus. Growth regulation through a β-adrenergic receptor can also be demonstrated in the culture model. The growth response mediated through a β-adrenergic receptor differs in several respects from that transduced through an al adrenergic receptor.
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    Molecular and cellular biochemistry 172 (1997), S. 37-46 
    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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  • 90
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    Molecular and cellular biochemistry 172 (1997), S. 111-120 
    ISSN: 1573-4919
    Keywords: differential display ; cardiac development ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract An estimated 15,000 different mRNA species are expressed in a typical mammalian cell. The differential expression of mRNAs in both a temporal and cell-specific manner determines the fate of the cell and creates the organism. Analysis of this differential gene expression has become a central aim of many laboratories attempting to understand the mechanisms underlying various biological processes. Currently, we are using a technique called differential display to analyze the differential expression of genes in cardiomyocytes. Differential display is a rapid and powerful technique that was introduced by Liang and Pardee in 1992. Since that time, it has been successfully applied by several groups, and it is quickly becoming a standard method for studying differential gene expression. Here, we present a detailed article discussing the differential display methodology and how we have utilized it to identify potential genes involved in cardiomyocyte proliferation. Furthermore, we have provided a list of materials and supplied examples of data obtained, in an effort to allow the reader to perform the technique with success in their own laboratory.
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  • 91
    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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  • 92
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    Molecular and cellular biochemistry 178 (1998), S. 157-162 
    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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  • 93
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    Molecular and cellular biochemistry 162 (1996), S. 51-58 
    ISSN: 1573-4919
    Keywords: metabolism ; glucose transporter ; adipocytes ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We tested the hypothesis that the constitutive glucose transporter (GLUT 1) in 3T3-L 1 adipocytes belongs to the family of glucose-regulated proteins which are transcriptionally regulated by glucose deprivation. Using cDNA probes for both GRP78 (BiP) and GLUT1, we show that the level of GRP78 mRNA increased by 15-fold within 24 h of glucose deprivation with little change in GLUT1 mRNA. The elevated GRP78 mRNA in turn led to a time-dependent increase in GRP78 protein. While glucose deprivation did not alter the expression of the normal glycoform of GLUT 1, a lower molecular weight glycoform accumulated with extended deprivation. Mannose and fructose, but not galactose, prevented the induction of GRP78 and accumulation of the abnormal GLUT1. Because GRP78 acts as a chaperone in other cell systems, we also sought evidence to support this activity in 3T3-L1 adipocytes. Using the technique of co-immunoprecipitation, we demonstrate that GRP78 bound several proteins unique to the glucose-deprived state. No deprivation-specific proteins could be detected in association with GLUT 1. These data lead us to conclude that GLUTl does not display characteristics of the glucose-regulated proteins, at least in 3T3-L1 adipocytes, a widely used model for differentiation, hormone action, and nutrient control. However, the mechanisms for activating traditional members of this family appear intact.
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  • 94
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    Molecular and cellular biochemistry 103 (1991), S. 41-50 
    ISSN: 1573-4919
    Keywords: insulin-like growth factor-1 ; binding proteins ; diabetes ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary Multiple factors contribute to the growth retardation which is a characteristic feature of uncontrolled diabetes. In this report we have examined the effects of streptozotocin-induced (STZ) diabetes on expression of insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding protein-1 (IGFBP-1) in various tissues. As early as 7 days after STZ administration there was a modest reduction in IGF-I mRNA abundance. The reduction (10–30%) was of similar magnitude in each of the 7 tissues examined; liver, kidney, lung, diaphragm, quadraceps, heart and adipose tissue. However, the reduction achieved statistical significance only in the lung (p 〈 0.05) and diaphragm (p 〈 0.01). A further reduction in IGF-I mRNA abundance was seen in many tissues, 32 and 91 days after STZ administration. In contrast to the decrease in IGF-I mRNA, IGFBP-1 mRNA was significantly increased in the liver and kidney of diabetic rats. IGFBP-1 mRNA was detectable at only very low levels in other tissues but was increased in diabetic rats compared non-diabetic rats. In diabetic rats, a highly significant correlation (R = 0.75, p 〈 0.001) between hepatic IGFBP-1 mRNA and glucose was observed whereas there was no significant correlation between serum glucose and hepatic IGF-I mRNA abundance (R = 0.24, p = NS). Treatment of diabetic rats with insulin resulted in a small, non significant increase in hepatic and renal IGF-I mRNA and a significant decrease in renal IGFBP-1 mRNA abundance. The observations reported here are consistent with the hypothesis that diminished IGF-I expression and inhibition of available IGF-1 by increased levels of IGFBP-1 may explain the impaired growth seen in diabetic animals.
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  • 95
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    Molecular and cellular biochemistry 186 (1998), S. 43-51 
    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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  • 96
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    Molecular and cellular biochemistry 201 (1999), S. 111-123 
    ISSN: 1573-4919
    Keywords: complement factor I ; TPA ; protein kinase C ; gene expression ; Hep G2 cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract This study examined the role of the protein kinase C (PKC) signalling pathway in the regulation of expression of human complement factor I (CFI) gene. The production of CFI by Hep G2 cells was enhanced in a dose- and time-dependent fashion by 12-O-tetradecanoyl-1,2-phorbol 13-acetate (TPA), a potent PKC activator. 4α-phorbol didecanoate, an inactive phorbol ester, had no effect on CFI synthesis. The TPA-dependent increase in CFI secretion was correlated with an increase in CFI mRNA levels. Forskolin, a cAMP-inducing agent, augmented the TPA response. W7, an inhibitor of protein kinase A and genistein, an inhibitor of protein tyrosine kinase(s) both did not prevent the increase in CFI expression mediated by TPA. However, calphostin C, a specific inhibitor of PKC, abolished the TPA-induced increase in CFI mRNA levels. Down regulation of intracellular PKC levels by prior exposure of Hep G2 cells to a high concentration of TPA also blocked the increase in CFI mRNA levels induced by TPA suggesting that the TPA effects were mediated via activation of PKC. mRNA decay studies indicated that the half-life of CFI mRNA in TPA-induced cells was not significantly different from control. Nuclear run-on transcriptional assays on the other hand demonstrated that whereas the CFI gene is transcribed under basal conditions in Hep G2 cells, TPA induced a 3-4 fold increase in the transcription rate of CFI gene in 24 h. The transcription rate of GAPDH gene did not change, indicating that the effects were not general on gene transcription. Transient transfections of Hep G2 cells with chloramphenicol acetyltransferase reporter gene (CAT) constructs containing a series of sequential 5′ deletions of the CFI promoter and CAT assays showed that the sequence between -136 and -130, containing an AP-1 consensus sequence (TGAGTCA) was required for the TPA response. This observation was substantiated by the finding that mutation of this AP-1 site to TttaTCA or TtAtcCA abolished the TPA responsiveness. The enhancement of the activity of transfected chimeric CAT constructs by TPA was abrogated by calphostin C and by pyrrolidine dithiocarbamate (an inhibitor of NF-κB and AP-1 transactivation). These results indicate that TPA regulation of CFI gene requires PKC signalling and is mediated by via a TPA response element (TRE) in the CFI promoter region located at -136/-130 and involves the transactivation of AP-1 and NF-κB transcription factors. We suggest that PKC may be one of the intracellular pathways that control CFI gene expression and that cellular processes (involving growth factors, hormones, cytokines etc.) that activate PKC may upregulate the expression of the CFI gene.
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  • 97
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium-binding protein ; gene expression ; Ca2+-ATPase ; brain microsomes ; aging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The expression of calcium-binding protein regucalcin and its effect on the microsomal Ca2+-ATPase activity in rat brain tissues was investigated. The expression of regucalcin mRNA was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) analysis in brain tissues using rat regucalcin-specific primers. Regucalcin concentration in the brain tissues was about 5 × 10-9 M as measured using enzyme-linked immunoadsorbent assay (ELISA), and this level was lowered with increasing age (50 weeks old). The presence of regucalcin (10-9 to 10-7 M) in the enzyme reaction mixture caused a significant decrease in Ca2+-ATPase activity in the brain microsomes of young rats (5 weeks old). Meanwhile, the enzyme activity was not significantly altered by the addition of calmodulin (1 or 50 μg/ml), calbindin (1 or 10 μg/ml), and S-100 A protein (5 or 25 μg/ml), which are other Ca2+-binding proteins in rat brain. The effect of regucalcin to inhibit microsomal Ca2+-ATPase activity was weakened in the brain of rats with increasing age (50 weeks old). The present study demonstrates that regucalcin is expressed in the brain, and that it can uniquely inhibit Ca2+-ATPase activity in the brain microsomes of rats. The findings suggest that regucalcin plays a role in the regulation of microsomal Ca2+-ATPase activity in rat brain tissues.
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  • 98
    ISSN: 1573-1561
    Keywords: Coleoptera ; Staphylinidae ; Tenebrionidae ; larvae ; defensive glands ; quinone ; naphthoquinone ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The abdominal glands of three bark-inhabiting larvae of generaBolitochara, Leptusa (Staphylinidae), andHypophloeus (Tenebrionidae) were studied chemically and morphologically. Behavior of the larvae indicated that secretion is emitted only after severe disturbance of the larvae. These mechanical contacts may also occur incidentally with coinhabiting nonpredatory arthropods when the beetle larvae move within small interstices under bark. Depending on the species, the secretions contained 1,4-benzoquinone derivatives and three 6-alkyl-naphthoquinones dissolved within various alkanes, alkenes, ethyl-, isopropyl-, and isoamylesters. More erratically distributed gland constituents also detected were acetophenone, benzyl propionate, and methyl hydroxybenzoate. In the laboratory, synthetic quinone-containing solutions simulating those found inLeptusa andBolitochara larvae acted as strong topical irritants and caused further damage to last-stageCalliphora vomitoria larvae if hydrocarbons or esters were used as solvents. The natural secretions ofHypophloeus versipellis elicited considerable mortality in two subcortical sciarid larvae cooccurring with tenebrionid larvae. Bioassay and secretion chemistry of the Staphylinidae/Tenebrionidae larval secretions indicated that they are typical defensive secretions that act topically. Morphological data characterized Bolitocharini larvae as possessing protuberant abdominal tergites supplied with an interiorly situated gland reservoir. After mechanical contact, the defensive secretion is topically applied to other arthropods by dabbing this tergal protuberance on targets. The defensive gland ofHypophloeus versipellis is unusual in possessing a movable reservoir opening situated at the anterior border of tergite IX. By this peculiar gland morphologyHypophloeus larvae are capable of shooting secretion droplets frontally from their slightly depressed dorsal abdominal surface without bending their abdominal tips dorsally. This seems an adaptation to the interstitial habitat of the larvae. The types of defensive glands and their phylogenetic value in Aleocharinae/Tenebrionidae larvae are discussed.
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  • 99
    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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    Molecular and cellular biochemistry 151 (1995), S. 55-60 
    ISSN: 1573-4919
    Keywords: regucalcin ; calcium ; gene expression ; kidney damage ; rat kidney cortex
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The alteration of Ca2+-binding protein regucalcin mRNA expression in the kidney cortex of rats administered cisplatin and cephaloridine, which can induce kidney damage, was investigated. Cisplatin (0.25, 0.5 and 1.0 mg/100 g body weight) or cephaloridine (25, 50 and 100 mg/100 g) was intraperitoneally administered in rats, and 1, 2 and 3 days later they were sacrificed. The alteration in serum findings after the administration of cisplatin (1.0 mg/100 g) or cephaloridine (50 and 100 mg/100 g) demonstrated chemically induced kidney damage; blood urea nitrogen (BUN) concentration increased markedly and serum inorganic phosphorus or calcium concentration decreased significantly. Moreover, the administration of cisplatin (1.0 mg/100 g) or cephaloridine (100 mg/100 g) caused a remarkable increase of calcium content in the kidney cortex of rats, indicating kidney damage. The expression of regucalcin mRNA in the kidney cortex was markedly reduced by the administration of cisplatin or cephaloridine in rats, when the mRNA levels were analyzed by Northern blotting using rat liver regucalcin cDNA (0.9 kb). The mRNA decreases were seen with the used lowest dose of cisplatin or cephaloridine. The present study clearly demonstrates that the mRNA expression of Ca2+-binding protein regucalcin in the kidney cortex of rats is decreased by chemically induced kidney damage.
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