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B Widegren

Publications and source records attributed to B Widegren.

31 records · Page 2Linked to original sources

The complete nucleotide sequence of the mitochondrial DNA of the fin whale, Balaenoptera physalus.

The composition of the mitochondrial DNA (mtDNA) of the fin whale, Balaenoptera physalus, was determined. The length of the molecule is 16,398 bp, and its organization conforms with that of other mammals. The general similarity between the mtDNA of the fin whale and the cow is greater than the similarity between the fin whale and other species (human, mouse, rat) in which the composition of the entire molecule has been described. The D-loop region of the mtDNA of the fin whale is 81% identical to the D-loop of dolphin DNA, and the central portion of the D-loop is similar to the bovine D-loop. The accumulation of transversions and gaps in the 12S and 16S rRNA genes was assessed by comparing the fin whale, cow, and human. The sequence difference between human and the whale and human and the cow was at the same level, indicating that the rate of evolution of the mtDNA rRNA genes is about the same in artiodactyls and cetaceans. In the 12S rRNA gene an accumulation rate of 0.05% per million years places the separation of cetaceans and artiodactyls at about 55 million years ago. The corresponding figure for human and either the whale or the cow is about 80 million years. In the 16S rRNA gene a 0.08% accumulation rate of transversions and gaps per million years yields concurring figures. A comparison between the cytochrome b gene of the fin whale and cytochrome b sequences in the literature, including dolphin (Stenella) sequences, identified the cetaceans as monophyletic and the artiodactyls as their closest relatives. The comparison between the cytochrome b sequences of the fin whale and Stenella showed that differences in codon positions one or two were frequently associated with a change in another codon position.

Amino Acid Sequence↗

Composition and chromosomal localization of cetacean highly repetitive DNA with special reference to the blue whale, Balaenoptera musculus.

Three highly repetitive DNA components--the common cetacean component, the heavy (GC-rich) satellite and the light (AT-rich) satellite--were were studied in the blue whale. Consensus sequences of the common component and the heavy satellite were determined on the basis of three repeats of the common component and eight repeats of the heavy satellite. The tandemly organized common cetacean component, which comprises a large portion of all cetacean--both odontocete (toothed whale) and mysticete (whalebone whale)--genomes has a repeat length of 1,760 bp and the three clones analysed showed a high degree of conformity. The repeat contains a 72 bp sequence with dyad symmetry and striking intrastrand complementarity. The rest of the repeat comprises a unique sequence. The repeat unit of the heavy satellite of the blue whale is 422 bp. Also this component is tandemly organized. About half the length of the repeat constitutes a unique sequence and the other half is made up of subrepeats with TTAGGG as a frequent motif. The light satellite has not been sequenced and its basic repeat unit has not yet been identified. The chromosomal localization of the three components was determined by in situ hybridization using 3H-labelled cloned fragments as probes. The common cetacean component was located in most interstitial and terminal C-bands. The heavy satellite occurred primarily in terminal C-bands. When the two components hybridized to the same terminal C-bands, the localization of the heavy satellite was distal to that of the common cetacean component. Neither component shared localization with the light satellite which is located in centromeric C-bands in just a few chromosome pairs.

Animals↗

The cytotoxic eosinophil cationic protein (ECP) has ribonuclease activity.

The eosinophil cationic protein (ECP) is a specific cytotoxic constituent of granules. In this work we demonstrated that ECP has a ribonuclease activity. Purified ECP was resolved by ion exchange chromatography into subfractions, which all showed ribonuclease activity. Another eosinophil granule protein, EPX, identical with eosinophil-derived neurotoxin (EDN) had a 125-fold higher RNase activity than ECP. ECP may exert its cytotoxic effects on parasites and cells because of its extreme basicity alone or it may be internalized and act by degrading mRNA.

Blood Proteins↗

DNA cloning and hybridization in deer species supporting the chromosome field theory.

The Cervidae show the largest variation in chromosome number found within any mammalian family. The eight species of deer which are the subject of this study vary in chromosome number from 2n = 70 to 2n = 6. Three species of Bovidae are also included since they belong to a closely related family. Digestion of nuclear DNAs with the restriction endonucleases Hae III, Hpa II, Msp I, Eco RI, Xba I, Pst I and Bam HI reveals that there is a series of highly repetitive sequences forming similar band patterns in the different species. There are two bands (1100 and 550 base pairs) which are common to all species although the two families separated more than 40 million years ago. To obtain information on the degree of homology among these conserved sequences we isolated a Bam HI restriction fragment of approximately 770 base pairs from red deer DNA. This sequence was 32P labeled and hybridized by the Southern blot technique with DNAs cleaved with Bam HI, Eco RI, Hpa II and Msp I. Moreover, the same sequence was cloned in the plasmid vector pBR322 nick translated with 32P and hybridized with the DNAs of 8 species of Cervidae and 3 of Bovidae. The same cloned probe was labeled with 3H and hybridized in situ with the metaphase chromosomes of red deer (2n = 68) and Muntiacus muntjak (2n = 7 male). Homologies are still present between the highly repetitive sequences of the 8 species of Cervidae despite the drastic reorganization that led to extreme chromosome numbers. Moreover, the cloned DNA sequence was found to occupy the same position, in the proximal regions of the arms, in both red deer (2n = 68) and M. muntjak (2n = 7 male) chromosomes. The ribosomal RNA genes and the centromeres in these species have also maintained their main territory despite the drastic chromosome reorganization. These results are experimental confirmation of the chromosome field theory which predicted that each DNA sequence has an optimal territory within the centromere-telomere field and tends to occupy this same territory following chromosome reorganization.

Animals↗

Characteristics of a conserved 1,579-bp highly repetitive component in the killer whale, Orcinus orca.

A tandemly organized, highly repetitive DNA component of the killer whale was sequenced. The length of the repeat was 1,579 bp. This unit, which characterizes all delphinids, shows stringent hybridization homology with a 1,740-bp repeat that is characteristic of all other cetacean families. The 1,579-bp component comprises approximately 15% of the killer-whale genome, in which it is repeated 4-5 X 10(5) times. Computer analysis of the sequence showed no linear repetition within the component. This indicates that the 1,579-bp unit has not evolved by amplification of shorter repeats. Several inverted repeats of substantial length were found in the 1,579-bp unit. The most conspicuous of these was a 72-bp sequence that deviated from matching in only three positions. The 72-bp sequence occurs within an open reading frame 330 bp in length. Transcriptional activity was registered in the cloned repeat in a cell-free system. The length of the transcript was approximately 340 nucleotides. The chromosomal localization of the 1,579-bp repeat was determined by in situ hybridization. The repeat was present in eight of 21 autosomal pairs and was found in almost all C-band-positive (constitutive heterochromatin) regions of the karyotype.

Animals↗

Conservation of highly repetitive DNA in cetaceans.

It is controversial whether odontocetes (toothed whales) and mysticetes (whalebone whales) have a common ancestry. Cetacean karyological uniformity, which is unique among mammalian orders, suggests a monophyletic origin; however, several anatomical authorities have maintained that odontocetes and mysticetes are diphyletic. We investigated the issue using Southern blot hybridization. Two labelled restriction fragment probes from the DNA of the sei whale (a mysticete) were hybridized to restricted DNA of cetacean species representing all extant families except the Eschrichtiidae, the gray whales. The probes hybridized to specific restriction fragments in all odontocete and mysticete materials. Hybridization showed preservation of hybridization homologies and a striking conservation of the length of highly repeated DNA sequences. The results are compatible with a common ancestry of odontocetes and mysticetes.

Animals↗

Conservation of repetitive DNA sequences in deer species studied by southern blot transfer.

The Cervidae show one of the largest variations in chromosome number found within a mammalian family. The five species of the deer family which are the subject of this study vary in chromosome number from 2n = 70 to 2n = 6. Digestion with the restriction enzymes EcoRI, HpaII, HaeIII and MspI reveals that there is a series of highly repetitive sequences forming similar band patterns in the different species. To obtain information on the degree of homology among these conserved sequences we isolated a HpaII restriction fragment of approximately 990 base pairs from reindeer DNA. This DNA sequence was 32P-labelled and hybridized by the Southern blot technique to DNAs cleaved with HpaII and HaeIII from the reindeer and four other Cervidae species. Hybridization to specific restriction fragments was recorded in all species. The patterns of hybridization showed a higher degree of similarity between reindeer, elk and roe deer than between reindeer and the Asiatic species (fallow deer and muntjac). Homologies are still present between the highly repetitive sequences of the five species despite the drastic reorganization that led to a change in chromosome number from 6 to 70.

Animals↗

In vivo and in vitro ethylene oxide exposure of human lymphocytes assessed by chemical stimulation of unscheduled DNA synthesis.

Factory workers exposed to ethylene oxide (EO), 0.5-1.0 ppm in factory air, together with matched controls from the same factory, were examined for evidence of toxic exposure by measurement of unscheduled DNA synthesis (UDS) induced by N-acetoxy-2-acetylaminofluorene (NA-AAF) and of chromosome aberrations in peripheral lymphocytes. The total chromatid gaps plus breaks were significantly elevated and NA-AAF-induced UDS was significantly reduced in the EO-exposed group as compared with the unexposed control group. The NA-AAF-induced UDS values negatively correlated to the duration (yr) of EO exposure (r = -0.45, p less than 0.02) and the number of chromosome breaks (r = -0.61, p less than 0.05), indicating an inhibition in vivo of DNA-repair capacity by EO. These data were verified in vitro by biochemical and autoradiographic studies of EO-induced UDS in human blood cells. Above 2 mM EO, UDS was inhibited in lymphocytes whether they were cultured for 24 or 122 h after alkylation with EO. Even at the subtoxic EO dose of 0.1 mM, lymphocytes were sensitized to additional exposures of NA-AAF, so that cytotoxicity was increased to 40% compared with 5% for the controls even though UDS was unaffected. It is concluded that EO was toxic to lymphocytes, even when they were sensitized at non-toxic EO doses to the cytotoxic action of other mutagens (e.g. NA-AAF), and the cells that did survive above 2 mM EO were inhibited in their DNA-repair capacity as judged by reduced UDS.

Adult↗

Regression of intracerebral rat glioma isografts by therapeutic subcutaneous immunization with interferon-gamma, interleukin-7, or B7-1-transfected tumor cells.

Progress in the definition of the roles of various costimulators and cytokines in determining the type and height of immune responses has made it important to explore genetically altered tumor cells expressing such molecules for therapeutic immunizations. We have studied the effect of therapeutic subcutaneous (s.c.) immunizations on the growth of preexisting intracerebral brain tumor isografts in the rat. Transfectant glioma cell clones expressing either rat interferon-gamma (IFN-gamma), rat interleukin-7 (IL-7), or rat B7-1 were selected. After irradiation (80 Gy) the clones were used for immunization (administered in up to four s.c. doses in a hind leg over 14-day intervals starting 1 day after the intracranial isografting of the parental tumor). Significant growth inhibition of the intracerebral parental tumors was induced by transfectants expressing IFN-gamma and IL-7, respectively. The strongest effect was observed with IFN-gamma-expressing cells, resulting in cures in 37% of the males and in 100% of the females. Immunization with IL-7 had a similar, strong initial effect, with significantly prolonged survival in the majority of the rats but a lower final cure rate (survival for >150 days). The B7-1-expressing tumor clones induced cures in seven of eight female rats; however, no cures were seen in the male rats. It was also shown that the B7-1-expressing cells were themselves strongly immunogenic in female rats, requiring high cell numbers to result in a progressively growing tumor upon s.c. isografting; this was not the case in male rats. As a whole, the results imply that despite the unfavorable location of intracerebral tumors, therapeutic s.c. immunizations with certain types of genetically altered tumor cells can induce complete regressions with permanent survival and without gross neurological or other apparent signs of brain damage. The present results demonstrate complete regressions when immunizations are initiated shortly after intracranial isografting, when the intracerebral tumor is small.

Animals↗