Search PubMed⌕ Search

Biomedical subjects

K Wolf

Publications and source records attributed to K Wolf.

At least 181 records · Page 10Linked to original sources
PubMed1986

Isolation and genetical and biochemical characterization of mutants resistant to the alkaloid lycorine.

Mutants resistant to 200 micrograms/ml of the alkaloid lycorine (LYCR) in non-fermentable substrate were isolated after nitrosoguanidine mutagenesis. Tetrad analysis and growth of heterozygous (LYCR/lycS) diploids from two different mutants revealed that a single nuclear and dominant mutation is responsible for the resistant phenotype. In the wild type total protein synthesis is only slightly inhibited, whereas DNA and RNA synthesis is lowered to about 30% of the control. In the lycorine resistant mutants all macromolecular syntheses are unaffected by the drug.

Amaryllidaceae Alkaloids↗
PubMed1986

[Capsule fibrosis in silicone implants].

The contracture of soft tissue capsule, appearing after implantation of all types of breast prosthesis, is the central problem of interest in the augmented breast. Clinical results were graduated in the Baker classification according to the degree of severity. Frequency rate depends on time lapse between implantation and post-examination, definition of capsular contracture, number of all patients and type of augmentation. Various factors are assumed to be causes of capsular contracture; i.e. haematoma, foreign body reaction, subclinical infections and physical-geometrical characteristics of prosthesis. It is possible to influence fibrosis with steroids and antibiotics.

Breast↗
PubMed1986

The mosaic cox1 gene in the mitochondrial genome of Schizosaccharomyces pombe: minimal structural requirements and evolution of group I introns.

The gene encoding subunit 1 of cytochrome oxidase (cox1) in the fission yeast Schizosaccharomyces pombe is polymorphic. In strain 50 it contains two group I introns with open reading frames (ORFs) in phase with the upstream exons (Lang, 1984). In strain EF1 two additional very short group I introns which do not possess ORFs were detected by DNA sequencing. These two introns (AI2a and AI3) share distinct characteristics concerning their nucleotide sequence and secondary structure and are located at identical positions as the introns AI4 and AI5 beta, respectively, in the cox1 gene of Saccharomyces cerevisiae. The sequence homology of the cob and cox1 genes around the splice points of introns AI2a, AI4, and BI4 (cob intron 4) might reflect horizontal gene transfer between the distantly related species S. pombe and S. cerevisiae.

Amino Acid Sequence↗
PubMed1985

The mitochondrial genome of the fission yeast Schizosaccharomyces pombe. 7. Continuous gene for apocytochrome b in strain EF1 (CBS 356) and sequence variation in the region of intron insertion in strain ade 7-50h.

The third BamHI fragment, containing most of gene for apocytochrome b, has been cloned and sequenced in the Schizosaccharomyces pombe strain EF1 (CBS 356). In contrast to strain ade 7-50h- (50) from the Leupold collection, in which the gene is interrupted by an intron of group II (Lang et al. 1984), the homologous gene in strain EF1 is continuous. This demonstrates that the intron in the gene for apocytochrome b is optional. Aligning the EF1 sequence with the homologous regions in strain 50, 2 base pair changes were found in the leader and 14 in the coding region. These changes led to 12 altered triplets, but 9 of them specify the same amino acid. Seven base changes were clustered within a stretch of 30 base pairs in the region in which the intron is inserted in strain 50. Five out of the resulting six triplet changes were also silent. These sequence variations around the highly conserved splice point region may be linked to the insertion or excision of the intron.

Ascomycota↗
PubMed1985

Cholinesterase activity and exposure time to acetylcholine as factors influencing the muscarinic inhibition of [3H]-noradrenaline overflow from guinea-pig isolated atria.

Guinea-pig isolated atria were incubated and loaded with [3H]-noradrenaline. The release of 3H and of [3H]-noradrenaline was induced by field stimulation (6-9 trains of 150 pulses at 5 Hz). The stimulation-evoked overflows of 3H and of [3H]-noradrenaline were determined. In the absence of an inhibitor of acetylcholinesterase, acetylcholine (12 min preincubation before nerve stimulation, up to 10 microM) failed to inhibit the evoked [3H]-noradrenaline overflow. In the presence of atropine, an increase by acetylcholine of evoked release was observed in the same atria. In contrast, the selective muscarinic agonist methacholine significantly decreased the evoked overflow. The inhibition was antagonized by atropine. Methacholine did not enhance release in the presence of atropine. When present for only 2 min, acetylcholine 10 microM inhibited the evoked overflow and no facilitation of release was observed in the presence of atropine. In the presence of physostigmine, acetylcholine (12 min preincubation, 1 and 10 microM) inhibited evoked [3H]-noradrenaline overflow, but the overflow was increased by acetylcholine 10 microM in the presence of atropine. In the presence of cocaine, corticosterone, phentolamine, propranolol and hexamethonium together, acetylcholine 1 microM inhibited the evoked [3H]-noradrenaline overflow. The inhibition was significantly enhanced in the presence of physostigmine. It decreased with preincubation time of the agonist, despite the presence of physostigmine and constant replacement by new drug. Neither inhibition nor facilitation of evoked release was observed in the presence of atropine. It is concluded that a muscarinic inhibition by acetylcholine (upon prolonged exposure time) may be masked by a concomitant facilitation of release and/or desensitization of the muscarinic inhibitory mechanism. Furthermore, degradation by acetylcholinesterase contributes in part to the ineffectiveness of acetylcholine as a presynaptic inhibitor. When a distortion of the overflow/release ratio was excluded, adrenergic and nicotinic effects were prevented, and acetylcholinesterase was inhibited, the fading of muscarinic inhibition by acetylcholine may have been exclusively due to a slow and moderate desensitization of the presynaptic muscarinic mechanism.

Acetylcholine↗
PubMed1985

Enhancement of the antitumor efficacy of lomustine by the radiosensitizer RSU 1069.

Previous investigations have shown that combining the radiation sensitizer misonidazole with conventional alkylating chemotherapeutic agents can lead to a therapeutic advantage. More recently, another sensitizer, RSU 1069, has been reported to give an enhancement of antitumor agent efficacy similar to that observed with misonidazole, but at an approximately tenfold lower sensitizer dose. One chemotherapeutic agent whose activity has been modified by sensitizers to a greater extent in tumors than in critical normal tissues is the nitrosourea lomustine (CCNU). The present studies evaluated the therapeutic benefit of combining RSU 1069 and CCNU in KHT sarcoma-bearing C3H/HeJ mice. The drugs were administered ip, and tumor response was assessed by measuring the survival of clonogenic KHT cells 22-24 hours after treatment. Normal tissue toxicity was determined using peripheral wbc counts 3 days after treatment and a 30-day lethality assay. Combining CCNU with a 0.38-mmol/kg dose of RSU 1069 increased tumor cell killing by a factor of approximately 1.9. Wbc toxicity and 30-day animal lethality increased with CCNU dose, but the addition of RSU 1069 enhanced either endpoint only slightly (factor of 1.0-1.2). The addition of RSU 1069 to CCNU treatment, therefore, led to a significant therapeutic benefit.

Animals↗
PubMed1985

In vivo interaction between radiation and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea in the absence or presence of misonidazole in mice.

The effect of combining the radiosensitizer misonidazole (MISO) with 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) prior to radiation on the response of various KHT sarcoma cell subpopulations was evaluated. Centrifugal elutriation was used to obtain homogeneous populations of cells with respect to their cell cycle position from suspensions prepared directly from treated solid KHT tumors. Survival in these subpopulations was determined using an in vivo to in vitro cloning assay. In situ treatment consisted of either (a) CCNU (3.0 mg/kg) plus MISO (1.0 mg/g), (b) radiation (15 Gy) alone, or (c) CCNU plus MISO preceding the radiation by 24 hr. In the latter protocol, one treatment effectively eliminated those cells being preferentially spared by the other. The CCNU-MISO followed by radiation combination not only reduced the treatment resistance due to hypoxia and cell cycle position but also left all cell subpopulations in the tumor equally sensitive to the treatment. To determine the nature of the interaction between these agents in detail, the extent of cell killing following such a treatment regimen was evaluated using isoeffect plot (isobologram) analysis. KHT sarcoma-bearing C3H mice were treated with various doses of CCNU either alone or in simultaneous combination with a 1.0-mg/g dose of MISO 24 hr prior to receiving a range of radiation doses. Combining CCNU and radiation led to significantly enhanced tumor cell killing. When evaluated by isoeffect plot analysis, the data points resulting from this combination fell well below the envelope of additivity, indicating that a supraadditive interaction between these two agents had occurred in the tumor cells. An even greater interaction was observed in the KHT sarcomas when MISO was combined with CCNU prior to irradiation. The findings of effective elimination of treatment-resistant subpopulations along with a supraadditive tumor cell kill offer, at least in part, an explanation for the previously described therapeutic advantage observed for the CCNU-MISO-radiation protocol.

Animals↗
PubMed1984

The mitochondrial genome of the fission yeast Schizosaccharomyces pombe. 2. Localization of genes by interspecific hybridization in strain ade7-50h- and cloning of the genome in small fragments.

A series of 18 small overlapping restriction fragments has been cloned, covering the complete mitochondrial genome of Schizosaccharomyces pombe. By hybridizing mitochondrial gene probes from Saccharomyces cerevisiae and Neurospora crassa with restriction fragments of Schizosaccharomyces pombe mitochondrial DNA, the following homologous genes were localized on the mitochondrial genome of S. pombe: cob, cox1, cox2 and cox3, ATPase subunit 6 and 9 genes, the large rRNA gene and both types of open reading frames occurring in mitochondrial introns of various ascomycetes. The region of the genome, hybridizing with cob exon probes is separated by an intervening sequence of about 2500 bp, which is homologous with the first two introns of the cox1 gene in Saccharomyces cerevisiae (class II introns according to Michel et al. 1982). Similarly, in the cox1 homologous region, which covers about 4000 bp, two regions were detected hybridizing with class I intron probes, suggesting the existence of two cox1 introns in Schizosaccharomyces pombe. Hybridization with several specific exon probes with a determined order has revealed that cob, cox1, cox3 and the large rRNA gene are all transcribed from the same DNA strand. The low intensities of hybridization signals suggest a large evolutionary distance between Schizosaccharomyces pombe and Saccharomyces cerevisiae or Neurospora crassa mitochondrial genes. Considering the length of the mitochondrial DNA of Schizosaccharomyces pombe (about 19.4 kbp) and the expected length of the localized genes and intron sequences there is enough space left for encoding the expected set of tRNAs and the small rRNA gene. The existence of leader-, trailer-, ori- and spacer sequences or further unassigned reading frames is then restricted to a total length of about 3000 bp only.

Ascomycota↗
PubMed1984

The mitochondrial genome of the fission yeast Schizosaccharomyces pombe. 3. Gene mapping in strain EF1 (CBS 356) and analysis of hybrids between the strains EF1 and ade7-50h-.

The Schizosaccharomyces pombe strain EF1 (CBS 356) is haploid, prototrophic, respiratory competent, and of homothallic mating type. From restriction enzyme analysis the length of the mitochondrial genome is 17.3 kilobase pairs, which is in good agreement with the value of 17.1 kilobase pairs determined by electron microscopy. The mitochondrial genome of strain EF1 is thus about 2.3 kilobase pairs shorter than that of strain ade7-50h- (about 19.4 kilobase pairs). A restriction map was constructed for 11 enzymes: For most, but not all of them, the pattern is nearly identical to that of strain ade7-50h-. The genes for the large ribosomal RNA, the subunits 1, 2, and 3 of cytochrome c oxidase, subunits 6 and 9 of ATP synthetase, and cytochrome b were localized by hybridization with mitochondrial DNA probes from Saccharomyces cerevisiae. The gene order was found to be the same in both yeast strains. From Southern hybridization of strain ade7-50h- with nick-translated mitochondrial DNA from strain EF1 it is evident that strain EF1 does not possess the intron, which is present in the cytochrome b gene of Schizosaccharomyces pombe strain ade7-50h-. Crosses between strain ade7-50h- and EF1 demonstrate that both the nuclear and the mitochondrial genomes are able to recombine. The mitochondrial genomes of 2 out of 30 independently isolated hybrids between the two strains are described as the result of recombination between the two parental mitochondrial genomes.

Adenosine Triphosphatases↗
PubMed1984

Potentiation of melphalan activity in the KHT sarcoma by the radiosensitizer RSU 1069.

The radiation sensitizer misonidazole (MISO) has been shown to potentiate the cytotoxic action of a variety of anti-cancer agents. Even larger enhancement ratios than those observed with MISO have been found with certain other nitroimidazoles. One agent reported to be particularly effective in combination with the chemotherapeutic agent melphalan is the sensitizer RSU 1069. The present studies therefore were designed to evaluate the effect of combining these two agents in the treatment of intramuscularly growing KHT sarcomas. Tumor response was assessed using an in vivo to in vitro clonogenic cell survival assay. When given at times ranging from 60 min before to 30 min after melphalan exposure, RSU 1069 was found to increase the tumoricidal activity of the chemotherapeutic agent. Complete dose response curves combining RSU 1069 and a range of melphalan doses then were determined. For comparison the effects of combining MISO or benznidazole (BENZO) with melphalan also were evaluated. All sensitizers were administered i.p. either 30 min before (BENZO) or simultaneously with (MISO, RSU 1069) the chemotherapeutic agent. Survival of clonogenic tumor cells assessed 22 to 24 hr after treatment was used to assay tumor response. When combined with melphalan, doses of RSU 1069 (0.38 mmol/kg), BENZO (0.3 mmol/kg) and MISO (5.0 mmol/kg) were found to yield dose modifying factors of 1.6, 1.5, and 1.4, respectively. These results indicate that potentiation of melphalan activity occurs at RSU 1069 doses which are approximately 10-fold lower than those of MISO, making this sensitizer as effective a potentiator of melphalan as so far tested in the KHT sarcoma.

Animals↗
PubMed1983

Expression of cloned mitochondrial DNA from the petite negative yeast Schizosaccharomyces pombe in E. coli minicells.

The minicell producing Escherichia coli strain D24 (lysogenic for phage lambda cI857) was transformed with the recombinant plasmid pDG3 containing the entire mitochondrial (mt) genome of the fission yeast Schizosaccharomyces pombe (S. pombe) cloned in the single BamHI-site of the E. coli plasmid pBR322 (Del Giudice 1981). By DNA-RNA hybridization it could be shown that the total mtDNA sequence of the plasmid pDG3 was transcribed in the E. coli minicells. The cloned mtDNA also directed the synthesis of at least five novel polypeptides with molecular weights between 7,200 and 34,000. When the minicell producing E. coli strain P678-54 was transformed with the hybrid plasmid pDG3, considerable portions of the inserted mtDNA sequences were deleted. One of the resulting plasmids (pDG4), lacking about two-thirds of the mtDNA sequence, directed the synthesis of new polypeptides in the range of 7,000 to 17,500 daltons. Another derivative of pDG3, the plasmid pDG5, containing one-sixth of the mtDNA sequence, directed the synthesis of at least three novel polypeptides. The mt origin of novel polypeptides coded by the hybrid plasmid pDG3 was demonstrated by use of antisera raised against total mitochondrial proteins from S. pombe and antisera against subunits II and III of cytochrome c oxidase from Saccharomyces cerevisiae (S. cer.).

Ascomycota↗
PubMed1983

In vivo potentiation of 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea by the radiation sensitizer benznidazole.

Recent studies in mouse tumor systems have indicated a potential therapeutic advantage in combining the radiosensitizer misonidazole (MISO) with cancer chemotherapy drugs. One agent the antitumor activity of which has been enhanced to a greater extent than its hematological or gastrointestinal toxicities is the nitrosourea, 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU). Recently, sensitizers more lipophylic than MISO have been reported to give greater tumor response enhancement when combined with CCNU. The present studies compared the potential therapeutic benefit of combining MISO (partition coefficient, 0.43) or benznidazole (BENZO) (partition coefficient, 8.5) in KHT sarcoma or RIF-1 tumor-bearing C3H mice. Both sensitizers were administered i.p. and given either 30 min before (BENZO) or simultaneously with (MISO) the chemotherapeutic agent. Survival of clonogenic tumor cells assessed 22 to 24 hr after treatment or in situ tumor growth delay were used as assays of tumor response. Normal tissue toxicity was determined using the drug dose yielding 50% animal lethality in 30 days end point. When combined with CCNU, doses of MISO (5.0 mmol/kg) or BENZO (0.3 mmol/kg) were found to yield approximately equivalent increases in both the tumor effect (enhancement ratio, approximately 1.8 to 2.0) and normal tissue toxicity (enhancement ratio approximately 1.3 to 1.4). Both sensitizers therefore led to a therapeutic benefit. However, although a approximately 10-fold lower dose of the more lipophylic sensitizer BENZO proved to be as effective as MISO at enhancing the tumoricidal effects of CCNU, this dose reduction did not result in a greater therapeutic gain for BENZO.

Animals↗
PubMed1982

Extrachromosomal inheritance of nalidixic acid resistance in the petite negative yeast Schizosaccharomyces pombe.

Spontaneous mutants resistant to nalidixic acid (NAL) were isolated from the petite negative yeast Schizosaccharomyces pombe (S. pombe). One of these mutants, resistant to 200 micrograms/ml NAL, nalr-Y13, was characterized both genetically and biochemically. The extrachromosomal inheritance of this mutation was demonstrated both by mitotic segregation and by mitotic haploidization analysis. In the wild-type, NAL at a concentration of 100 micrograms/ml almost completely inhibits incorporation of [14C]adenine in total DNA as well as in mitochondrial DNA. In the NAL-resistant mutant both total DNA synthesis and mitochondrial DNA synthesis were resistant to the drug. These results are discussed in view of previously published findings on the close interaction between the two DNA synthesizing systems in S. pombe.

Ascomycota↗