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U Bahr

Publications and source records attributed to U Bahr.

35 records · Page 2Linked to original sources
PubMed1998

Identification of a thymidylate synthase gene within the genome of Chilo iridescent virus.

The thymidylate synthase (TS, EC 2.1.1.45) is essential for the de novo synthesis of dTMP in pro- and eucaryotic organisms. Consequently it plays a major role in the replication of the DNA genome of a cell or a DNA virus. The gene encoding the TS of Chilo iridescent virus (CIV) was identified by nucleotide sequence analysis of the viral genome and was mapped within the EcoRI CIV DNA fragments G and R. Computer assisted analysis of the DNA nucleotide sequence between the genome coordinates 0.482 and 0.489 revealed an open reading frame (ORF) of 885 nucleotides. This ORF was found to encode a polypeptide of 295 amino acid residues (33.9 kDa) that showed significant homologies to known TS of different species including mammals, plants, fungi, protozoa, bacteria, and DNA viruses. The highest amino acid homologies were found between the CIV-TS and the TS of herpesvirus ateles (54.0%), Saccharomyces cerevisiae (51.8%), herpesvirus saimiri (51.0%), rhesus monkey rhadinovirus (50.7%), mouse (50.5%), rat (50.2%), varicella-zoster virus (50.2%), equine herpesvirus 2 (50.0%), and the human TS (48.4%). The CIV-TS contains six amino acid domains that are highly conserved in the TS of other species. Within these domains the major amino acid residues are present for which a functional role has been reported. The CIV-TS was found to be more closely related to the TS of eucaryotes than to the TS of procaryotes indicating the phylogenetic origin of the CIV-TS gene. The identification of a TS gene in the genome of CIV is the first report of a viral TS that is not encoded by a herpesvirus or a bacteriophage.

Amino Acid Sequence↗
PubMed1998

Identification and characterization of the Tupaia herpesvirus DNA polymerase gene.

Tupaia herpesviruses (THVs) have been isolated from malignant lymphomas and from degenerating lung or spleen cell cultures of tree shrews (Tupoia spp.), but because of a lack of genetic information the final classification of THVs is still open. In the present work the viral DNA polymerase (DPOL) gene was mapped within the genome of the different THV strains using PCR and degenerate oligonucleotide primers. Nucleotide sequences of the DPOL genes of THV strains 1 to 5 were determined and used for comparative analyses. The transcriptional activity of the THV-2 DPOL gene was confirmed by RT-PCR. It was found that the different THV strains are very closely related to each other. When compared to other herpesviruses the highest amino acid sequence identities detected were with DPOLs of the murine and human cytomegaloviruses. These results justify the conclusion that THVs are members of the subfamily Betaherpes-virinae.

Amino Acid Sequence↗
PubMed1997

High-sensitivity analysis of neutral underivatized oligosaccharides by nanoelectrospray mass spectrometry.

Nanoscale electrospray ionization (nano-ESI) overcomes the sensitivity problems found and reported for neutral oligosaccharides in conventional microscale forced-flow ESI. For a series of compounds ranging from trisaccharides to larger polymers with molecular masses up to 6 kDa, sample concentrations of 10(-5) M, i.e., 10 pmol total sample load, yielded very intense singly or multiply cationized molecule ions in an ion-trap mass spectrometer. In a dilution series, it is exemplified that, at the 10(-8) M level, molecule ion signals can be clearly registered with a S/N ratio of about 7. Only 100 amol of sample has been consumed in this experiment. Investigation of an oligosaccharide-peptide mixture revealed that the oligosaccharide is suppressed in conventional ESI, whereas in nano-ESI both analytes are detected at comparable and high intensities. Mechanistic implications are discussed, emphasizing the influence of surface activity for the two ESI techniques. The very low flow rates inherent to nano-ESI of about 30 nL/min, together with the high signal intensity, make it possible to fully employ the MSn capabilities of an ion-trap mass spectrometer for structural analysis. From less than 1 microL of sample solution it is possible to make consecutive fragmentation experiments up to MS7 to obtain valuable information about the structure of complex oligosaccharides.

Mass Spectrometry↗
PubMed1997

Delayed extraction time-of-flight MALDI mass spectrometry of proteins above 25,000 Da.

Matrix-assisted laser desorption/ionization (MALDI) with delayed extraction (DE) has been optimized for mass analysis of high-mass proteins and glycoproteins with masses above 25,000 Da. Under optimized experimental conditions, i.e. using a weak extraction field strength and a long delay time, a steep drop in mass resolution above 30,000 Da is no longer observed and an improvement of more than a factor of 10 is obtained compared with the non-DE case, at least up to 66 kDa in a 1.2 m time-of-flight mass analyzer. On this level of resolution the factors limiting further improvements become apparent, i.e. adduct ion formation between matrix and analyte, but also cationization and further non-matrix-related adducts, as well as prompt fragmentation. Moreover, heterogeneity of the sample is often the reason for the detection of broad signals for larger proteins. Within these limitations, DHBs (a 9:1 mixture of 2,5-dihydroxybenzoic acid and 2-hydroxy-5-methoxybenzoic acid) gave by far the best results.

Glycoproteins↗
PubMed1997

The DNA sequence of Chilo iridescent virus between the genome coordinates 0.101 and 0.391; similarities in coding strategy between insect and vertebrate iridoviruses.

Chilo iridescent virus (CIV), the type species of the genus Iridovirus within the family Iridoviridae, is highly pathogenic for larvae of important pest insects. The virions contain a single linear double-stranded DNA molecule (209 kbp) that is circularly permuted and terminally redundant. The nucleotide sequence of the viral genome between the genome coordinates 0.101 and 0.391 (60,170 bp) was determined by automated cycle sequencing. This particular region of the CIV genome contains 112 open reading frames (ORFs) with coding capacities for 50 to 1186 amino acids. The alignment of the deduced amino acid sequences with well-characterized proteins stored in protein databases led to the identification of several genes with significant homologies, such as the largest subunit of the DNA-dependent RNA polymerase, large subunit of the ribonucleoside-diphosphate reductase, endonuclease, protein-tyrosine phosphatase, helicase, global transactivator, two apoptosis inhibitor homologs, antibiotic peptide homolog, and others. The highest homologies were detected between putative viral gene products of CIV and the corresponding viral proteins of lymphocystis disease virus of fish (LCDV), which belongs to the genus Lymphocystivirus within the iridovirus family.

Amino Acid Sequence↗
PubMed1983

Cyclophosphamide levels in serum and spinal fluid of multiple sclerosis patients treated with immunosuppression.

The cyclophosphamide content of urine, serum and spinal fluid of immuno-suppressed multiple sclerosis patients was determined using a newly developed off-line combination of high pressure liquid chromatography and field desorption mass spectrometry. Although urinary excretion of the drug varied considerably between patients, the levels in serum and spinal fluid were in the same range, a relationship that may facilitate control of the dosage.

Adult↗
PubMed1982

Determination of lithium and rubidium in physiological fluids and tissues of rabbits during the reproductive phase.

1. The natural concentrations of lithium and rubidium were determined during the reproductive phase of rabbits by field desorption mass spectrometry. 2. Samples of serum, milk, amniotic fluid and placenta tissue were analysed. 3. The concentration changes in serum during the reproductive phase were between 2.61 and 5.02 micrograms/l for lithium and between 107.78 and 136.28 micrograms/l for rubidium. 4. Correlations between the concentration in maternal serum and bone growth of the fetus as well as the formation of milk were found. 5. Concentrations of 9.30 micrograms/l lithium and 1780.00 micrograms/l rubidium in the milk lead to the assumption that these trace metals are essential for the metabolism of the young rabbit.

Amniotic Fluid↗
PubMed1981

Isolation, identification and determination of cyclophosphamide and two of its metabolites in urine of a multiple sclerosis patient by high pressure liquid chromatography and field desorption mass spectrometry.

The off-line combination of high pressure liquid chromatography and field desorption mass spectrometry has been used for the simultaneous isolation, identification and determination of cyclophosphamide and two of its metabolites, 4-ketocyclophosphamide and carboxyphosphamide in urine from a patient suffering from multiple sclerosis. Cyclophosphamide and its metabolites were separated using reverse phase liquid chromatography. Field desorption mass spectrometry was employed for identification and quantification. The technique applied needs no derivatization for analysis. The limits of detection by field desorption mass spectrometry for 1, 2 and 3 are factor of about 4 X 10(3)-10(5) lower than those of a common variable ultraviolet detector. Quantitative determination was carried out using the method of stable isotope dilution with deuterated analogues of 1, 2 and 3. In a pilot study, the ratio of 1:2:3 was determined to 1:0.02:0.6. One ml of urine is sufficient for simultaneous analysis of the three compounds. The typical analysis time, including separation by liquid chromatography and field desorption measurement, is about 30 minutes.

Chromatography, High Pressure Liquid↗
PubMed1980

Determination of cyclophosphamide in urine, serum and cerebrospinal fluid of multiple sclerosis patients by field desorption mass spectrometry.

The levels of cyclophosphamide in samples of urine, serum and cerebrospinal fluid of multiple sclerosis patients, who had received the drug orally (4 x 100 mg/day) have been determined by field desorption mass spectrometry using the principle of stable isotope dilution. After thorough preparation of the samples, concentrations of cyclophosphamide of 10 to 60 micrograms/ml in urine and 200 to 400 ng/ml in serum and cerebrospinal fluid have been determined from 0.6-3 ml samples. The first quantitative data for cyclophosphamide in cerebrospinal fluid of multiple sclerosis patients could be established without the use of radioactive material. The relatively high level of the parent drug found in the fluid at the end of a 3-weeks treatment may shed some light on the mechanism of action of this drug in the treatment of the disease.

Cyclophosphamide↗
PubMed1980

Quantitative measurement of time-dependent thallium distribution in organs of mice by field desorption mass spectrometry.

The time-dependent distribution of the toxic heavy metal TI in mouse organs was determined after feeding 80, 130, and 160 mg/kg TI. Quantitative measurements were performed by field desorption mass spectrometry with stable isotope dilution. No pretreatment of the tissue samples, other than homogenizing and centrifugation, was necessary. The precision of the data was about +/- 10%. The main results are as follows: Heart, liver, kidney, and stomach showed an organ-specific initial uptake of TI during the first 2-3 h. This was followed by a period of washout to relatively low TI levels. Brain TI uptake was comparatively low and constant during the first 12 h. In the terminal stage (24 h) all organs, including the brain, contained increased TI levels of the same order of magnitude.

Animals↗
PubMed1978

Determination of lithium in microlitre amounts of human body fluids at therapeutic and normal levels by stable isotope dilution and field desorption mass spectrometry.

The use of field desorption mass spectrometry for the determination of lithium in body fluids at therapeutic levels--ppm region--as well as at the normal level--ppb region--has been developed. The use of a stable isotope enriched internal standard, together with the outstanding sensitivity of field desorption for alkali cations and the high specificity of mass spectrometry, allows a quantitative determination of lithium in microlitre amounts of body fluids, such as plasma, saliva and urine. The assay allows a determination of lithium even at ultratrace concentrations where routine spectroscopic procedures cannot be applied. Analysis of plasma required a simple protein precipitation, whereas saliva and urine could be analysed without treatment. The precision of the data obtained ranged from 2--10%. The time consumption for one analysis in routine work is about 20--30 min.

Humans↗
PubMedDate not supplied

Nano-electrospray ionization mass spectrometry: addressing analytical problems beyond routine.

The advent of nano-electrospray ionization (nano-ESI) has considerably extended the usability of ESI in the analytical mass spectrometric laboratory. One of the remarkable features of nano-ESI is its extremely low sample consumption. Only a few microliters of analyte solution (10(-5)-10(-8) M) are sufficient for molecular weight determination and structural investigations by MS/MS. But nano-ESI is more than just a minimized-flow ESI; the low solvent flow rate also affects the mechanism of ion formation. As a consequence, the area of ESI-MS applications is significantly enhanced. Oligosaccharides, glycosides as well as glycoproteins can be analyzed more easily than with normal ion spray. The same holds for the analysis of non-covalent complexes sprayed directly from aqueous solutions.

Animals↗
PubMedDate not supplied

Quantitative investigations on the diaplacental transfer of thallium by field desorption mass spectrometry.

The placental transfer of thallium cations in pregnant mice was investigated by determining the thallium concentrations in fetal and maternal tissue 0.5 to 24 h after application of thallium. The maternal dose of thallium was 8 mg/kg body weight throughout. Uterus and fetus were found not to differ from other organs like heart and liver in time course and magnitude of thallium uptake with an initial surge during the first few hours of exposure to thallium and a rapid decrease to steady 12 and 24 h values somewhat lower than those found in the kidney. Diaplacental transfer is therefore assumed comparatively rapid and a specific placental barrier for thallium does not seem to exist. For the determination of thallium concentrations Field Desorption Mass Spectrometry was utilized as a reliable, fast and sensitive method for the analysis of metal cations in biological material. This method does not require extensive pretreatment of the tissue and total sample amounts in the range of milligrams and less are sufficient for quantitative analysis.

Animals↗