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  • HPLC
  • Recombinant DNA
  • Springer  (61)
  • Elsevier
  • 1985-1989  (61)
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Year
  • 1
    Electronic Resource
    Electronic Resource
    Evidence for high molecular weight Na−Ca exchange in cardiac sarcolemmal vesicles (1988)
    Springer
    The journal of membrane biology 106 (1988), S. 211-218 
    Details
    ISSN: 1432-1424
    Keywords: Na−Ca exchange ; sarcolemma ; reconstitution ; HPLC ; target sizing ; cardiac
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Cardiac sarcolemma (SL) vesicles were subjected to irradiation inactivation-target sizing analyses and gel permeation high performance liquid chromatography (HPLC) to ascertain the weight range of native Na−Ca exchange. Frozen SL vesicle preparations were irradiated by electron bombardment and assayed for Na−Ca exchange activity. When applied to classical target sizing theory, the results yielded a minimum molecular weight (M r) of approximately 226,000±20,000sd (n=6). SL vesicle proteins were solubilized in 6% sodium cholate in the presence of exogenous phospholipid and fractionated by size on a TSK 30XL HPLC column. Eluted proteins were mixed 1∶1 with mobile phase buffer containing 50mg/ml soybean phospholipid and reconstituted by detergent dilution. The resulting proteoliposomes were assayed for Na−Ca exchange activity. Na−Ca exchange activity eluted in early fractions containing larger proteins as revealed by SDS-PAGE. Recovery of total protein and Na−Ca exchange activity were 91±7 and 68±11%, respectively. In the peak fraction, Na−Ca exchange specific activity increased two-to threefold compared to reconstituted controls. Compared to the elution profile of protein standards under identical column conditions, sodium cholate solubilized exchange activity had a minimumM r of 224,000 Da. Specific45Ca2+-binding SL proteins withM r of 234,000, 112,000, and 90,000 Da were detected by autoradiography of proteins transferred electrophoretically to nitrocellulose. These data suggest that native cardiac Na−Ca exchange is approximately 225,000 Da or larger. The exact identification and purification of cardiac Na−Ca exchange protein(s) remains incomplete.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01872159
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  • 2
    Electronic Resource
    Electronic Resource
    Determination of furosine as a measure for irreversibly bound glucose in human fibrinogen (1989)
    Springer
    Monatshefte für Chemie 120 (1989), S. 1147-1158 
    Details
    ISSN: 1434-4475
    Keywords: Human fibrinogen ; Furosine ; Glycation ; HPLC ; Nα-Formyl-Nɛ-(desoxy-1-D-fructosyl-1)-L-lysine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Description / Table of Contents: Zusammenfassung In der vorliegenden Arbeit wird ein Analysenverfahren vorgestellt, das eine Bestimmung der irreversiblen, nichtenzymatischen Glucosylierung von Fibrinogen aus menschlichem Blut ermöglicht. Die Methode beruht auf der Bestimmung von Furosin, welches während der Hydrolyse von Amadori-Produkten gebildet wird. Für die Validierung der erarbeiteten Furosin-Methode waren die Synthese von Nα-Formyl-Nɛ-(desoxy-1-D-fructosyl-1)-L-lysin als Modell und von Furosin als Eichstandard erforderlich. In Hydrolyseversuchen wurde das Ausmaß der Furosinbildung aus Fructoselysin und Fibrinogen untersucht. In Übereinstimmung mit der Literatur wurde gezeigt, daß es möglich ist, den Glucosylierungsgrad von Fibrinogen durch In-vitro-Inkubation mitD-Glucose zu erhöhen. Die Fibrinogengewinnung aus Blut erfolgte mit bekannten Techniken der Probenahme und Aufarbeitung. Durch Reduktions- (Reaktionsblindwert) und Hydrolyseschritte ist es möglich, Furosin flüssigkeitschromatographisch (UV-Detektion) quantitativ zu erfassen. Durch Anwendung dieses Analysenverfahrens kann das Ausmaß der Glucosylierung von menschlichem Fibrinogen mit guter Reproduzierbarkeit vergleichend erfaßt werden. Die Kalibrierung erfolgt gegen den synthetisierten Furosinstandard. Die praktische Durchführbarkeit wurde gezeigt.
    Notes: Summary An analytical procedure is presented which enables for the determination of irreversibly, nonenzymatic glycation of fibrinogen isolated from human blood. The method is based on determination of furosine, which is formed during hydrolysis of Amadori-products. For validation of the worked out furosine assay, synthesis of Nα-formyl-Nɛ-(desoxy-1-D-fructosyl-1)-Llysine as a model substance and furosine as a calibration standard were necessary. Several hydrolysis experiments showed the extent of furosine formation from fructosyllysine and from human fibrinogen. The increase of fibrinogen glycation by incubation withD-glucose could also be confirmed according to the literature. Using well-known techniques for sampling, work up and in performing reduction and hydrolysis steps, quantitative determination of furosine by high-performance liquid chromatography is possible. By application of the analytical assay, the extent of glycation of human fibrinogen can be analyzed with good precision. Calibration is performed by means of a prepared furosine standard. The practical handling of the method is shown.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00809298
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  • 3
    Electronic Resource
    Electronic Resource
    Trypsin in human milk (1986)
    Springer
    Cellular and molecular life sciences 42 (1986), S. 39-41 
    Details
    ISSN: 1420-9071
    Keywords: Human milk ; trypsin ; affinity chromatography ; HPLC ; radioimmunoassay
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Human milk trypsin was purified by adsorption chromatography on cellulose-bound 4-aminobenzamidine; its molecular weight was about 24,000 daltons. Its concentration determined by a radioimmunoassay varies between 2.9 and 5.6 μg/l.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01975884
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  • 4
    Electronic Resource
    Electronic Resource
    Makisterone A and 24-methylenecholesterol from the ovaries of the honey bee,Apis mellifera L (1986)
    Springer
    Cellular and molecular life sciences 42 (1986), S. 200-201 
    Details
    ISSN: 1420-9071
    Keywords: Ecdysteroids ; molting hormones ; makisterone A ; HPLC ; RIA ; honey bee ; ovaries ; Apis mellifera ; neutral sterols ; 24-methylenecholesterol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Makisterone A, a 28-carbon ecdysteroid (molting hormone) has been isolated from the ovaries of queen bees. Analysis by reversed-phase and silica high performance liquid chromatography (HPLC) in conjunction with a radioimmune assay (RIA) revealed about 11 ng of makisterone A present per gram of ovaries on a fresh weight basis. No C27 ecdysteroids were detected. The predominant neutral sterol present was 24-methylenecholesterol.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01952468
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  • 5
    Electronic Resource
    Electronic Resource
    Kinins in cerebrospinal fluid: Reduced concentration in spontaneously hypertensive rats (1986)
    Springer
    Cellular and molecular life sciences 42 (1986), S. 1238-1239 
    Details
    ISSN: 1420-9071
    Keywords: Kinins ; bradykinin ; kallidin ; cerebrospinal fluid ; HPLC ; hypertension
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rat cerebrospinal fluid contains peptides which displace radiolabeled bradykinin from its specific antibodies. Two peptides which showed the same retention time as kallidin and bradykinin in a reverse phase high pressure liquid chromatography system were detected in cerebrospinal fluid of rats. The concentration of radioimmunologically detected kinins in the cerebrospinal fluid of spontaneously hypertensive rats of the Okamoto strain was lower than that of the Wistar Kyoto control rats.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01946402
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  • 6
    Electronic Resource
    Electronic Resource
    Sporulation delay by stable cAMP analogues in the slime moldPhysarum polycephalum (1986)
    Springer
    Development genes and evolution 195 (1986), S. 276-280 
    Details
    ISSN: 1432-041X
    Keywords: cAMP-analogues ; Sporangia morphogenesis ; HPLC ; Microinjection ; Physarum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plasmodial cells of the slime moldPhysarum polycephalum become competent for sporulation following a prolonged period of starvation in darkness. Then sporulation can be induced by illumination. Microinjections of the stable (Sp)- and (Rp)-diastereoisomers of adenosine cyclic 3′,5′ monothionophosphate before and during a sensitive period from the start of illumination until 5 h after lead to a significant delay in the sporulation process. Both of the diastereoisomers of cyclic AMP prolong the time for sporangia to form in darkness. However, the (Sp)-diastereoisomer is more effective and causes morphological changes in plasmodia. The experimental data suggest that cyclic AMP is decisively involved in light-induced differentiation in the lower eukaryotoPhysarum polycephalum.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02438961
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  • 7
    Electronic Resource
    Electronic Resource
    Molecular cloning of a ribosomal protein gene from the fission yeast Schizosaccharomyces pombe (1986)
    Springer
    Current genetics 10 (1986), S. 365-370 
    Details
    ISSN: 1432-0983
    Keywords: Recombinant DNA ; Cross hybridization ; 2D-electrophoresis ; Hybrid selection translation ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Using the structural gene for the ribosomal protein L3 from Saccharomyces cerevisiae as a probe, we isolated a homologous fragment from genomic DNA of Schizosaccharomyces pombe. Analysis of the plasmid carrying this fragment by hybridization selection and 2D-electrophoresis revealed a 31 kDa ribosomal protein. Transformation of the vector pDB248x containing this fragment into Schizosaccharomyces pombe leads to an increased level of mRNA suggesting that we have cloned the entire and actively transcribed gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00418408
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  • 8
    Electronic Resource
    Electronic Resource
    Large mitochondrial genome and mitochondrial DNA size polymorphism in the mosquito parasite, Romanomermis culicivorax (1986)
    Springer
    Current genetics 11 (1986), S. 71-77 
    Details
    ISSN: 1432-0983
    Keywords: Mitochondrial DNA ; Population heterogeneity ; Molecular evolution ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Physical characterization of the mitochondrial genome derived from the obligate mosquito parasite, Romanomermis culicivorax has generated some surprising physical properties regarding the molecular structure of nematode mitochondrial DNA (mtDNA). Restriction enzyme analysis of this mtDNA has revealed a mitochondrial genome size of approximately 26 kb, the largest metazoan mtDNA reported to date. Isofemale lineages are monomorphic for one of three size variants, differing by 500-1,000 base pairs, present in our original field population. Cloned hybridization probes derived from a single region exhibiting a 600 by size polymorphism share strong homology with several spatially separated sites distributed about the mtDNA. This suggests that the homology is a result of repeated DNA sequence elements contained within this mitochondrial genome that contribute to mtDNA size polymorphism.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00389428
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  • 9
    Electronic Resource
    Electronic Resource
    The isolation of specific genes from the basidiomycete Schizophyllum commune (1987)
    Springer
    Current genetics 12 (1987), S. 547-554 
    Details
    ISSN: 1432-0983
    Keywords: Schizophyllum commune ; Transformation ; Gene isolation ; Basidiomycetes ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have developed a routine way to isolate genes directly from the basidiomycete fungus, Schizophyllum commune. Plasmid DNA from a genomic gene library was used to isolate five specific genes by complementation of Schizophyllum mutations via transformation. The mutant strains were deficient in the ability to synthesize either adenine (ade2 and ade5), uracil (ural, encoding orotidine-5′-phosphate decarboxylase; OMPdecase), tryptophan (rpl, encoding indole-3-glycerol phosphate synthetase; IGPS) or para aminobenzoic acid (pab1). In each case, Southern analysis revealed that transformation to prototrophy was concomitant with the integration of vector sequence into the genome of the S. commune mutant. Total DNA from transformants was restricted, religated, and used to transform E. coli. Ampicillin resistant plasmids were recovered from E. coli and tested for their ability to transform the corresponding mutant of S. commune. Plasmids complementing the ade2, adeS, pabl, trpl, and ural mutations were recovered.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419565
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  • 10
    Electronic Resource
    Electronic Resource
    Poly-cis carotene pathway in the Scenedesmus mutant C-6D (1988)
    Springer
    Archives of microbiology 150 (1988), S. 590-594 
    Details
    ISSN: 1432-072X
    Keywords: Acyclic poly-cis carotenes ; Carotene biosynthesis ; HPLC ; Pigment mutant C-6D ; Scenedesmus obliquus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The mutation of Scenedesmus obliquus strain C-6D is expressed in the dark only. Under these conditions, this strain synthesizes acyclic poly-cis carotenes. Cyclic carotenoids like β-carotene or xanthophylls are absent. In the light the normal cyclic carotenes and xanthophylls are synthesized in the all-trans configuration. The poly-cis carotencs present in dark cultures have been analysed and quantitated. It could be shown that these poly-cis carotenes are identical with the poly-cis carotenes synthesized by the tomato mutant Lycopersicon esculentum var. ‘Tangella’. The poly-cis pathways, however, are different regulated in the two organisms. The tomato mutant accumulates prolycopene as the major carotene, whereas the mutant C-6D accumulates mainly pro-ζ-carotene. Furthermore, the mutation in ‘Tangella’ is constitutive in light in contrast to Scenedesmus C-6D. Besides that, Scenedesmus C-6D synthesizes a further cis-carotene isomer of phytofluene as well as of ζ-carotene. The configuration of these carotenes still have to be elucidated. The occurrence of this poly-cis carotene biosynthetic pathway by a mutation of only one enzyme, the phytoene desaturase which, however, is only expressed in darkness under heterotrophic conditions, is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00408255
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  • 11
    Electronic Resource
    Electronic Resource
    The simultaneous determination of muramic acid and glucosamine in soil by high-performance liquid chromatography with precolumn fluorescence derivatization (1988)
    Springer
    Biology and fertility of soils 6 (1988), S. 125-130 
    Details
    ISSN: 1432-0789
    Keywords: Muramic acid ; Glucosamine ; Soil ; HPLC ; Biomass
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The optimal release and quantitative estimation of muramic acid and glucosamine were studied simultaneously in soil samples. The effect of hydrolysis conditions, HCl concentration, hydrolysis time, the ratio of soil dry weight to acid, and the recovery of reference substances were investigated. Derivatization of the fluorogenic reagent o-phthalaldehyde, in the presence of 2-mercaptoethanol with the residue of a soil hydrolysate, was achieved by optimizing the relative amounts of o-phthalaldehyde to hydrolysate in the reaction mixture, the pH of both, and the incubation period. A linear relationship was found between the fluorescence response and the concentration of the test substances. The muramic acid, as well as the glucosamine (o-phthalaldehyde) derivatives gave single peaks, and complete separation from interfering substances at the picomol level was achieved in a short time (3 h preparation and 30 min for chromatography) by using high-performance liquid chromatography.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00257661
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  • 12
    Electronic Resource
    Electronic Resource
    Drug monitoring of quinine by HPLC in cerebral malaria with acute renal failure treated by haemofiltration (1987)
    Springer
    European journal of clinical pharmacology 33 (1987), S. 293-296 
    Details
    ISSN: 1432-1041
    Keywords: falciparum malaria ; quinine ; acute renal failure ; HPLC ; haemofiltration ; plasma levels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The monitoring of quinine by HPLC in 3 patients suffering from cerebral malaria with acute renal failure and treated by haemofiltration is reported. The recommended dose of quinine in this situation is reduced to 10 to 15 mg·kg−1·day−1. However, in the first patient, when given quinine 10 mg kg−1·day−1 the plasma concentration was mainly below the recommended therapeutic range of 5 to 15 mg/l. In consequence, the dose of quinine in the second patient was elevated to quinine dihydrochloride 15.1 mg·kg−1·day−1 which produced plasma concentrations in the low therapeutic range. In the third patient, an unreduced dose of quinine dihydrochloride 25.7 mg·kg−1·day−1 was employed, resulting in plasma concentrations above 15 mg/l, which is generally assumed to be toxic, although, no sign of acute quinine toxicity was seen. The antimalarial effect in all three patients was satisfactory. Quinine was estimated in the haemofiltrate in two patients and was found to be below the limit of sensitivity (0.25 mg/l). Plasma quinine did not change during or shortly after haemofiltration. It is concluded that in case of acute renal failure in cerebral malaria the dose of quinine should be reduced, but that the common recommendation of 10 to 15 mg·kg−1·day−1 may be too low, and that haemofiltration has no marked influence on the total body clearance of quinine.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00637565
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  • 13
    Electronic Resource
    Electronic Resource
    Influence of smoking and gender on the disposition kinetics of metoprolol (1987)
    Springer
    European journal of clinical pharmacology 33 (1987), S. 355-361 
    Details
    ISSN: 1432-1041
    Keywords: metoprolol ; smoking ; gender ; pharmacokinetics ; HPLC ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The purpose of this study was to examine the influence of cigarette smoking and gender on the pharmacokinetics of metoprolol. Eighteen volunteers with no evidence of clinical disease each randomly received the following doses of metoprolol tartrate: 100 mg orally, 200 mg orally and 20 mg as a constant-rate intravenous infusion over 20 min. The only significant difference between smokers (S) and nonsmokers (NS) was that S had a larger steady-state volume of distribution (3.3 vs 2.5 l/kg). There were no differences in half-life, systemic clearance or bioavailability (f). No differences were observed between males (M) and females (FM) for any of the kinetic parameters examined. Systemic bioavailability varied markedly between subjects (range: 15 to 92%). In fifteen of the eighteen subjects, f was higher after the 200-mg dose compared to the 100-mg dose. These results suggest that metoprolol may be subject to saturable presystemic elimination and extend the previous observations of Johnsson et al. [1] who showed that f increased from 31% to 46% when doses were increased from 20 to 100 mg. However, the difference in f as the dose is increased is unlikely to be clinically significant since the mean difference is smaller than the variation in f among subjects.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00637630
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  • 14
    Electronic Resource
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    Disposition of monodesethylamodiaquine after a single oral dose of amodiaquine and three regimens for prophylaxis againstPlasmodium falciparum malaria (1987)
    Springer
    European journal of clinical pharmacology 33 (1987), S. 409-414 
    Details
    ISSN: 1432-1041
    Keywords: amodiaquine ; Plasmodium falciparum malaria ; monodesethylamodiaquine ; HPLC ; pharmacokinetics ; prophylaxis ; metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The disposition of monodesethylamodiaquine was studied in four healthy subjects after a single oral dose of 10 mg/kg amodiaquine base. Amodiaquine was not found in any sample, but the major metabolite monodesethylamodiaquine was detected and was assumed to be the sole derivative that contributed significantly to antimalarial activity in the blood. The best fit for the decay of the metabolite was obtained with a three-compartment model. The half-lives of the first two phases were 3.2 to 11.4 h for t1/2α1 and 22.7 to 50.3 h for t1/2α2 in plasma. The half-life of the terminal phase ( t1/2β) was between 9 and 18.2 days. The concentration in whole blood was 4- to 6-times higher than in plasma. Three schedules (alternate days, weekly, daily) of the conventional prophylactic dose of 10 mg/kg per week were compared in six other healthy subjects. There were significant differences in the plasma monodesethylamodiaquine levels between the three schedules.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00637639
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  • 15
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    Fermentation of recombinant yeast producing hepatitis B surface antigen (1987)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1987), S. 117-121 
    Details
    ISSN: 1476-5535
    Keywords: Fermentation ; Recombinant DNA ; Hepatitis B surface antigen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Fermentations were performed to determine parameters affecting the expression of hepatitis B surface antigen (HBsAg) in the yeastSaccharomyces cerevisiae containing the HBsAg gene. These studies emphasized inereasing both the relative abundance (HBsAg: cell mass) and total production of HBsAg. Specific activity was increased 70-fold when cells were grown in shake flasks containing nonselective rather than selective medium. The addition of adenine, ammonium sulfate or glucose to the complex medium reduced the production of antigen. Results similar to those achieved in shake flasks were obtained when the growth was performed in fermenters. A nutrient addition system was employed to increase the production of cells and HBsAg. The addition of glucose to the culture medium increased cell mass 6-fold but decreased the production of antigen. This imbalance was corrected by supplementing the glucose with complex nutrients.
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    URL: http://dx.doi.org/10.1007/BF01569510
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  • 16
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    Aufnahme und Verteilung von14C-Retinylacetat in Organe und Sinnesgewebe (Innenohr) des Meerschweinchens (1985)
    Springer
    European journal of nutrition 24 (1985), S. 187-196 
    Details
    ISSN: 1436-6215
    Keywords: 14C-Retinylacetat ; Sinneszellgewebe ; Vitamin-A-Verteilung ; HPLC ; Autoradiographie
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Summary The uptake and chemical identification of14C-retinyl acetate in the inner ear of the guinea-pig after oral administration is reported. For methodological reasons the experiment was carried out in vitamin A-deficient guinea-pigs. In the sensory tissues a time-dependent distribution was found similar to that in other organs. The chemical identification shows that the orally administered labeled retinyl acetate can be detected as retinyl palmitate in the membranous structures of the inner ear. This may be an indication for the ability of the inner ear tissues to esterize and probably store the transport form of vitamin A, retinol.
    Notes: Zusammenfassung Es wird über die Aufnahme und chemische Identifizierung von14C-Retinylacetat in das Innenohr des Meerschweinchens nach per oraler Verabreichung berichtet. Die Untersuchung wurde aus methodischen Gründen bei Vitamin-A-mangelernährten Tieren durchgeführt. Es findet sich eine zeitabhängige Verteilung in den Sinnesgeweben, die der anderer Organe gleicht. Die chemische Identifizierung zeigt, daß das als Retinylacetat verabreichte markierte Vitamin A in den membranösen Strukturen des Innenohres in Form des Retinylpalmitats zu finden ist. Dies mag ein Hinweis sein auf die Fähigkeit des Innenohrgewebes, die Transportform des Vitamins A, das Retinol, zu verestern und eventuell auch zu speichern.
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    URL: http://dx.doi.org/10.1007/BF02019356
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  • 17
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    Characterization of seed storage proteins of urdbean (Vigna mungo) (1988)
    Springer
    Plant foods for human nutrition 38 (1988), S. 163-173 
    Details
    ISSN: 1573-9104
    Keywords: urdbean ; protein fractions ; amino acid composition ; HPLC ; SDS-PAGE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Dehulled and defatted flour of urdbean (Vigna mungo), Var T-9, contained 25% protein with maximum contribution by globulins (63%). Albumins and glutelins contributed 12% and 21% respectively, whereas prolamins were present only in traces (1%). Globulins were further fractionated into legumin and vicilin type proteins which were present in the ratio of 4:1. All the protein fractions were heterogenous in nature as revealed by high performance liquid chromatography. SDS-polyacrylamide gel electrophoresis revealed the total protein sample to contain 21 different components with molecular weights ranging from 8.92 to 117.49kd. Albumins, globulins, prolamins and glutelins resolved into 4, 8, 6 and 13 different sized components of molecular weights ranging from 10.23 to 25.53, 10.84 to 112.72, 10.33 to 51.52 and 8.91 to 112.72kd, respectively. Amino acid analysis of all fractions revealed that glutamic acid was present in maximum concentration followed by aspartic acid and lysine. Just like other pulse proteins, the urdbean proteins were also deficient in sulphur containing amino acids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01091721
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  • 18
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    Screening for organic acids in fish tissues with special reference to the distribution of taurine inRutilus rutilus L. (1987)
    Springer
    Fish physiology and biochemistry 3 (1987), S. 145-149 
    Details
    ISSN: 1573-5168
    Keywords: Rutilus rutilus L. ; cyprinids ; organic acids ; taurine ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A liquid chromatographic method for rapid profiling of some organic acids and other compounds of physiological interest in fish tissues is presented. The method provides the detection of α-ketoglutaric, malic and pyruvic acid and is optimized for the quantitation of citric, phosphoric, succinic, lactic and fumaric acids, as well as glucose and especially taurine. These substances were separated in a single analytical run on a cation exchange column with dilute sulfuric acid as the elutant. The effluent was monitored by a refractive index or an UV detector at 208 nm wavelength. Resolution was satisfactory. No further treatment was found to be necessary when samples were deproteinized with perchloric acid. Taurine (2-aminoethansulfonic acid), not previously described in the roach (Rutilus rutilus L.), occurs in considerable amounts in red and white muscle and heart tissue. However, it is also present in all other tissues.
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    URL: http://dx.doi.org/10.1007/BF02180416
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  • 19
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    Studies on the tocopherol status in blood serum of premature babies and infants (1986)
    Springer
    European journal of nutrition 25 (1986), S. 1-8 
    Details
    ISSN: 1436-6215
    Keywords: Vitamin E ; tocopherol ; premature infant ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition , Medicine
    Description / Table of Contents: Zusammenfassung Bei 88 Frühgeborenen im Alter von 0 bis 2 Jahren wurde der Gehalt an natürlich vorkommendem Tocopherol im Blutserum durch Hochleistungsflüssigkeitschromatographie (HPLC) mit fluorimetrischer Detektion bestimmt. In 11 Fällen wurde während eines längeren Zeitraums der Tocopherolstatus von Patienten, die bekannte Vitamin-E-Präparate oral und/oder parenteral aufnahmen, untersucht. Es wurde keine Korrelation zwischen alpha-Tocopherol-Spiegel und Gestationsalter bzw. Geburtsgewicht beobachtet. Alle Frühgeborenen, die Vitamin-E-Präparate erhielten, wiesen im Durchschnitt Serum-Tocopherolgehalte von mehr als 0,5 mg/100 ml auf. Die höchste alpha-Tocopherolkonzentration, die während einer Vitamin-E-Therapie gemessen wurde, war 3,28 mg/100 ml. Eine Infusion von Intralipid, einem Produkt, das aus fraktioniertem Sojaöl erhalten wird, verursachte einen signifikanten Anstieg der gamma- und delta-Tocopherolgehalte im Blutserum. Für die Halbwertszeit von delta-Tocopherol im Serum wurde ein Wert von etwa 24 Stunden berechnet.
    Notes: Summary The level of naturally occurring tocopherols in blood serum of 88 preterm infants, aged from birth to 2 years, was determined by the high performance liquid chromatographic (HPLC) method using fluorescence detection. In 11 cases, patients were assayed for their tocopherol status in longitudinal studies, receiving known amounts of vitamin E supplements orally and/or parenterally. No correlation was found between serum alpha-tocopherol level and gestational age nor birth weight. All preterm infants receiving the vitamin E preparation showed an average serum tocopherol content of more than 0.5 mg/100 ml. The highest alpha-tocopherol concentration registered during vitamin E therapy was 3.28 mg/100 ml. Infusion of Intralipid, a product derived from fractionated soybean oil, caused a significant increase of gamma- and delta-tocopherol in blood serum. The half-life of delta-tocopherol in serum was calculated to be about 24 hours.
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    URL: http://dx.doi.org/10.1007/BF02023615
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    Factors affecting the expression of foreign proteins inEscherichia coli (1987)
    Springer
    Journal of industrial microbiology and biotechnology 1 (1987), S. 277-282 
    Details
    ISSN: 1476-5535
    Keywords: Recombinant DNA ; Gene expression ; Genetic engineering ; Biotechnology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A variety of factors affect the expression of foreign proteins inEscherichia coli. These include: promoter strength, efficiency of ribosome binding, stability of the foreign protein inE. coli, location of the foreign protein inE. coli, the codons used to encode the foreign protein, the metabolic state of the cell, and the location, stability and copy number of the foreign gene. This paper contains a critical review of these factors with the idea that a detailed understanding of them is the key to the development of strategies for the efficient large-scale production of foreign proteins inE. coli.
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    URL: http://dx.doi.org/10.1007/BF01569305
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    Refractometric determination of sulphate and other anions in plants separated by High-Performance Liquid Chromatography (1986)
    Springer
    Plant and soil 91 (1986), S. 129-132 
    Details
    ISSN: 1573-5036
    Keywords: Anion Determination ; Chloride ; HPLC ; Nitrate ; Nitrite ; Phosphate ; Refractometric detection ; Selenate ; Sulphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Anion exchange chromatography by High Performance Liquid Chromatography (HPLC) and the refractometric detection of the separated anions proved to be a rapid and sensitive method for the simultaneous determination of sulphate, nitrate, nitrite, chloride and phosphate in plants. Anion content was quantified using sodium selenate as internal standard. Only 20 μl of a ten times diluted deproteinized plant extract was needed for determination.
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    URL: http://dx.doi.org/10.1007/BF02181825
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    Opioid mechanisms in insects, with special attention toLeucophaea maderae (1988)
    Springer
    Cellular and molecular neurobiology 8 (1988), S. 269-284 
    Details
    ISSN: 1573-6830
    Keywords: neuroendocrine mechanisms ; evolutionary history ; neuropeptides ; endogenous opioids ; immunocytochemistry ; HPLC ; opioid receptors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. This review article provides information on the evolutionary history of neuroendocrine and related regulatory mechanisms. It focuses on the presence, diverse roles, and modes of operation of one class of neuropeptides, the endogenous opioids, in insects. 2. Opioid peptides, closely resembling those of vertebrates, have been identified in the brain and related neuroendocrine structures by means of immunocytochemistry and high-pressure liquid chromatography. 3. The demonstration of naloxone-sensitive, high-affinity binding sites for Met-enkephalin-like neuropeptides in the brain and digestive tract ofLeucophaea deserves special attention because it provides new insights into the functional significance of opiate receptors paralleling those known in vertebrates. 4. Possible roles of receptor-mediated opioid systems in the insects discussed are regulation of the cyclicity of the female reproductive system, maintenance of normal midgut function mediated by the recurrent nerve, and locomotor activity.
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    URL: http://dx.doi.org/10.1007/BF00711169
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    Atrial natriuretic peptide in the central nervous system of the rat (1988)
    Springer
    Cellular and molecular neurobiology 8 (1988), S. 339-391 
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    ISSN: 1573-6830
    Keywords: atrial natriuretic peptide ; central nervous system ; immunocytochemistry ; HPLC ; radioimmunoassay ; autoradiography ; intracerebral injection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. Studies of the presence of atrial natriuretic peptide immunoreactivity and receptor binding sites in the central nervous system have revealed unusual sites of interest. 2. As a result, numerous studies have appeared that indicate that brain atrial natriuretic peptide is implicated in the regulation of blood pressure, fluid and sodium balance, cerebral blood flow, brain microcirculation, blood-brain barrier function, and cerebrospinal fluid production. 3. Alteration of the atrial natriuretic peptide system in the brain could have important implications in hypertensive disease and disorders of water balance in the central nervous system.
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    URL: http://dx.doi.org/10.1007/BF00711224
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    Synthesis of stereoisomers of 8-methyl-2-decanol and esters attractive to severalDiabrotica sp. (1985)
    Springer
    Journal of chemical ecology 11 (1985), S. 1371-1387 
    Details
    ISSN: 1573-1561
    Keywords: Asymmetric synthesis ; stereoisomers ; sex pheromone ; Diabrotica ; Coleoptera ; Chrysomelidae ; (R)-2-methylbutyric acid ; HPLC ; diastereomers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Details of the syntheses of the four stereoisomers of 8-methyl-2-decanol and its propanoate ester are given. The racemic ester, two of its stereoisomers, and one stereoisomer as an acetate are attractive to several species ofDiabrotica. The key steps in the syntheses involve high-performance liquid chromatograpic resolutions of diastereomers to achieve high configurational enrichment of each site and generation of (R)-2-methylbutyric acid by chemical degradation ofd-isoleucine.
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    URL: http://dx.doi.org/10.1007/BF01012138
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    HPLC separation and wavelength area ratios of more than 50 phenolic acids and flavonoids (1985)
    Springer
    Journal of chemical ecology 11 (1985), S. 383-395 
    Details
    ISSN: 1573-1561
    Keywords: HPLC ; retention time ; area ratio ; phenolic acids ; flavonoids ; soybean ; Glycine max
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Relative retention times and wavelength area ratios for over 50 standard compounds were calculated using reverse-phase HPLC. The standard compounds analyzed included benzoic acids, cinnamic acids, benzene carboxylic acids, acetic acids, coumarins, benzaldehydes and a variety of flavonoid compounds including flavanones, flavones, isoflavones, and their glycosides. Each standard compound was chromatographed by three different gradient elutions. Compounds were detected by UV absorption at 254 nm and 280 nm. Relative retention times with respect to two different internal references and the 254nm: 280nm wavelength area ratio was determined for each standard. Soybean root and seed extracts were analyzed for the presence of the standard compounds using the chromatographic conditions described.
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    URL: http://dx.doi.org/10.1007/BF01411424
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    Paecilotoxin production in clinical or terrestrial isolates of Paecilomyces lilacinus strains (1989)
    Springer
    Mycopathologia 108 (1989), S. 195-199 
    Details
    ISSN: 1573-0832
    Keywords: Paecilomyces lilacinus ; paecilotoxin ; distribution ; HPLC ; clinical isolates ; entomopathogenic ; nematode-killing fungi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The production of paecilotoxin from various isolates of Paecilomyces lilacinus was studied using three different media and high performance liquid chromatography (HPLC). Alkaline medium was found to be suitable for the production of the toxins. Among 20 strains tested, 19 including four clinical isolates were found to produce the toxins. Production patterns of paecilotoxins were very similar in each strain and the main toxins were A and B.
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    URL: http://dx.doi.org/10.1007/BF00436225
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    Accumulation of a peptide toxin from the cyanobacterium Oscillatoria agardhii in the freshwater mussel Anadonta cygnea (1989)
    Springer
    Hydrobiologia 183 (1989), S. 211-216 
    Details
    ISSN: 1573-5117
    Keywords: Cyanobacteria ; freshwater mussels ; toxin accumulation ; tissue distribution ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Swan mussels (Anodonta cygnea) were exposed to a toxic strain of the cyanobacterium Oscillatoria agardhii. Mussels accumulated large amounts of the peptide Oscillatoria toxin which was present in low concentrations within the cyanobacterial cells in the test aquaria (40–60 µg Oscillatoria toxin/1). The toxin concentration in the mussels increased during the experiment and after 15 days of exposure the concentration was 70 ± 2 µg/g freeze dried tissue (mean ± range of values). The highest concentration of the toxin (130 µg/g of freeze dried tissue) was found in the hepatopancreatic tissue. The toxin did not seem to be metabolized in the mussels and they were not killed by the high toxin concentrations within them. After two months in clean water still detectable amounts of toxin were present in the mussels.
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    URL: http://dx.doi.org/10.1007/BF00007241
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    Molecular cloning of the plant plasma membrane H+-ATPase (1987)
    Springer
    Plant and soil 99 (1987), S. 185-196 
    Details
    ISSN: 1573-5036
    Keywords: Antibody ; H+-ATPase ; Membrane ; Recombinant DNA ; Transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Mineral transport across the plasma membrane of plant cells is controlled by an electrochemical gradient of protons. This gradient is generated by an ATP-consuming enzyme in the membrane known as a proton pump, or H+-ATPase. The protein has a catalytic subunit of Mr=100,000 and is a prominent band when plasma membrane proteins are analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We generated specific rabbit polyclonal antibody against the Mr=100,000 H+-ATPase and used the antibody to screen λgtll expression vector libraries of plant DNA. Several phage clones producing immunoreactive protein, and presumably containing DNA sequences for the ATPase structural gene, were isolated and purified from a carrot cDNA library and a Arabidopsis genomic DNA library. These studies represent our first efforts at cloning the structural gene for a plant plasma membrane transport protein. Applicability of the technique to other transport protein genes and the potential for use of recombinant DNA technology in plant mineral transport research are discussed.
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    URL: http://dx.doi.org/10.1007/BF02370165
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    Determination of plant growth substances in liquid endosperm of immature walnut (Juglans nigra) nuts by an ELISA technique (1988)
    Springer
    Plant growth regulation 7 (1988), S. 209-215 
    Details
    ISSN: 1573-5087
    Keywords: Walnut ; Juglans nigra ; Endosperm ; IAA ; ABA ; Cytokinin ; GA3 ; HPLC ; ELISA immunoenzymatic test ; Antibody
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Plant growth substances (PGSs) were analysed in liquid endosperm of black walnut using HPLC and an ELISA procedure. Of all the PGSs studied, we show no GA3, low levels of cytokinins (io6A, i6Ade, i6Ado) and ABA, and very high level of IAA
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    Analysis of fish bile with HPLC — fluorescence to determine environmental exposure to benzo(a)pyrene (1989)
    Springer
    Hydrobiologia 188-189 (1989), S. 561-566 
    Details
    ISSN: 1573-5117
    Keywords: benzo(a)pyrene equivalents ; bile ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Brown bullhead from the Black River, Ohio, have a high incidence of liver neoplasia which is associated with elevated concentrations of polynuclear aromatic hydrocarbons (PAHs) in the sediment. We evaluated the use of biliary concentrations of benzo(a)pyrene [B(a)P] equivalents as a means for determining PAH exposure. Bile was collected from 16 brown bullheads and 8 common carp taken from each of two Lake Erie tributaries in Ohio, the industrialized Black River and the non-industrialized Old Woman Creek. Hatchery bullhead (n = 8) were used to determine base levels of PAHs. A high performance liquid chromatography (HPLC) — fluorescence technique was used to determine the concentration of B(a)P equivalents in the bile samples. The area of all peaks fluorescing at 380/430 nm was summed to give a single value for B(a)P equivalents in each sample. Concentrations of B(a)P equivalents generally reflected concentrations of PAH in sediment where fish were collected. Bile taken from Black River carp contained the highest concentration of B(a)P equivalents and was significantly different from all other groups. The value obtained for Black River bullhead was also high and was found to be significantly different from hatchery bullhead. B(a)P equivalents varied between carp and bullhead from the same habitat possibly because of differing food habits or metabolic pathways. However, our results indicate that relative levels of B(a)P equivalents in the bile of fish correspond well to B(a)P levels in sediment and may offer a means of determining environmental exposure of fish to the parent compound.
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    URL: http://dx.doi.org/10.1007/BF00027823
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    Protein and amino acids analysis during alimentary-agar extraction from Gelidium sesquipedale (1987)
    Springer
    Hydrobiologia 151-152 (1987), S. 513-522 
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    ISSN: 1573-5117
    Keywords: seaweed ; protein ; amino acid ; extraction ; Gelidium sesquipedale ; residue ; solubilization ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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    URL: http://dx.doi.org/10.1007/BF00046176
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    Effects of carbohydrates and nitrogen on the development of anthocyanins of a red leaf peach (Prunus persica (L.) Batsch) in vitro (1987)
    Springer
    Plant cell, tissue and organ culture 9 (1987), S. 103-110 
    Details
    ISSN: 1573-5044
    Keywords: cyanidin 3-glucoside ; HPLC ; Prunus persica ; carbohydrate ; nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Heterozygous red leaf peach (Prunus persica (L.) Batsch) shoots were implanted on media with varying nitrogen and carbohydrate regimes to identify a combination which elicited maximum anthocyanin production in explants. A medium with relatively low nitrogen (5 mM NH4+ and 10 mM NO3-) and high sucrose (234 mM) was most effective in stimulating anthocyanin production. Sucrose was more effective as a carbon source than glucose, fructose, or starch under given nitrogen levels. The major anthocyanin in red leaf peach was tentatively identified as cyanidin 3-glucoside based on PC and HPLC analysis.
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    Optical resolution of helical and planarchiral metacyclophanes by high performance liquid chromatography on (+)-poly(triphenylmethyl-methacrylate) (1985)
    Springer
    Journal of inclusion phenomena and macrocyclic chemistry 3 (1985), S. 51-54 
    Details
    ISSN: 1573-1111
    Keywords: Chiral recognition ; enantiomer separation ; helical and planarchiral molecules ; HPLC ; inclusion chromatography ; (+)-poly(triphenylmethyl-methacrylate)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Racemic helical and planarchiral ten-membered metacyclophanes1 to 7 are efficiently resolved into the enantiomers using high performance liquid chromatography on (+)-poly(triphenylmethyl-methacrylate).
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    The Saccharomyces cerevisiae MET3 gene: Nucleotide sequence and relationship of the 5′ non-coding region to that of MET25 (1987)
    Springer
    Molecular genetics and genomics 210 (1987), S. 307-313 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Methionine metabolism ; Negative control ; Regulatory regions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae, the expression of several genes implicated in methionine biosynthesis is coregulated by a specific negative control. To elucidate the molecular basis of this regulation, we have cloned two of these genes, MET3 and MET25. The sequence of MET25 has already been determined (Kerjan et al. 1986). Here, we report the nucleotide sequence of the MET3 gene along with its 5′ and 3′ flanking regions. Plasmids bearing different deletions upstream of the transcribed region of MET3 were constructed. They were introduced into yeast cells and tested for their ability to complement met3 mutations and to respond to regulation by exogenous methionine. The regulatory region was located within a 100 bp region. The sequence of this regulatory region was compared with that of MET25. A short common sequence which occurs 250–280 bp upstream of the translation initiation codon of the gene was found. This sequence is a good candidate for the cis-acting regulatory element.
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    A multigene family in Calothrix sp. PCC 7601 encodes phycocyanin, the major component of the cyanobacterial light-harvesting antenna (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 296-304 
    Details
    ISSN: 1617-4623
    Keywords: Complementary chromatic adaptation ; Gene expression ; Photoregulation ; Phycobilisome ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In cyanobacteria, light is harvested by phycobilisomes which are essentially made up of chromophoric proteins called phycobiliproteins. We have characterized two gene clusters (cpcB1, cpcA1 and cpcB3, cpcA3) each encoding the two subunits of phycocyanin (βPC and αPC, respectively), one of the major phycobiliproteins in Calothrix 7601. Downstream from the gene encoding the PCα subunit in cluster 1, an open reading frame was found, cpcE1. These genes are organized in two transcriptional units, namely: cpcB3 A3 and cpcB1 A1 E1. All these genes are transcribed whatever the chromatic light received during cell growth. Consequently, although only one type of “constitutive” PC has been biochemically characterized, we have demonstrated that there are two cpc operons “constitutively” transcribed in this strain. With the previously described red light “inducible” cpcB2 A2 operon, there are three copies of the PC encoding genes in Calothrix 7601. The significance of this newly described multigene family in cyanobacteria is discussed. We have also mapped the 5′ and 3′ termini of the major transcript from the cpc1 operon. Analysis of the 5′ untranslated region of this transcript has revealed alternative secondary structures which are proposed to play a role in the regulation of the expression of this operon.
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    Gene cloning in Aspergillus nidulans: isolation of the isocitrate lyase gene (acuD) (1986)
    Springer
    Molecular genetics and genomics 202 (1986), S. 271-275 
    Details
    ISSN: 1617-4623
    Keywords: Aspergillus ; Isocitrate lyase ; Cloning ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An Aspergillus nidulans gene library was constructed in a high-frequency transformation vector, pDJB3, based on the Neurospora crassa pyr4 gene. This gene library was used to isolate the structural gene for isocitrate lyase (acuD) by complementation of a deficiency mutation following transformation of A. nidulans. Plasmids rescued in Escherichia coli were able to transform five different A. nidulans acuD mutants. Transformation using plasmids containing the cloned fragment resulted in integration at the acuD locus in six of nine transformants.
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    Integration of foreign DNA following microinjection of tobacco mesophyll protoplasts (1986)
    Springer
    Molecular genetics and genomics 202 (1986), S. 179-185 
    Details
    ISSN: 1617-4623
    Keywords: Gene transfer ; Plant cell transformation ; Plant tissue culture ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary DNA from a bacterial plasmid containing the T-DNA border sequences of Agrobacterium tumefaciens was transferred into the nucleus or the cytoplasm of tobacco mesophyll protoplasts by microinjection. Following culture in hanging drops, some of these protoplasts produced calli containing the foreign DNA sequences. Evidence for the presence of the injected plasmid DNA in these calli was provided by Southern hybridization analysis. The results demonstrated that random portions of the bacterial plasmid were linked to plant DNA and that integration did not occur at the T-DNA borders present on the injected plasmid. The average number of integrated copies ranged from less than one to 1–2 per tobacco genome. The frequency of integration averaged 14% with intranuclear injections compared to 6% with cytoplasmic injections. With further refinement, the use of microinjection may allow the introduction of many different types of genetic elements into plants.
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    Cloning, sequencing and expression of a Bacillus bacteriolytic enzyme in Escherichia coli (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 241-248 
    Details
    ISSN: 1617-4623
    Keywords: Autolysin ; Hydrolase ; In vitro transcription and translation ; Recombinant DNA ; Transcriptional and translational signals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Several hundred bacterial isolates were screened for bacteriolytic activity by growing them on agar medium containing autoclaved, lyophilized Micrococcus lysodeikticus cells as the substrate. A Bacillus sp. producing the largest lytic zone was selected. A genomic bank of this selected bacterium was constructed in the multi-functional vector pTZ18R, with partial SauIIIA DNA fragments inserted at the SalI restriction site. Screening of 800 colonies of this bank for cell lysis gave 5 recombinants exhibiting lytic activity, as detected by analysis of extracts of sonicated Escherichia coli cells on denaturing polyacrylamide gels containing autoclaved, lyophilized M. lysodeikticus cells as the substrate. One clone (pBH2500), expressed inE. coli strain NM522, was found to code for a lytic enzyme corresponding, in molecular weight, to the 27 kDa Bacillus sp hydrolase. This clone with an insertion of 2.5 kb was then subcloned as a 929 bp EcoRI-SauIIIA fragment in pTZ18R (pBH929) and showed higher cell lytic activity. A unique open reading frame for a protein of 251 amino acids, followed by a putative terminator sequence, was found after a consensus ribosome binding site. A putative leader sequence was identified in the first 37 amino acids. One truncated subclone (pBH703), corresponding to 196 out of 251 residues from the protein N-terminal end, still possessed lytic activity.
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    Secretion and export of IGF-1 in Escherichia coli strain JM101 (1988)
    Springer
    Molecular genetics and genomics 215 (1988), S. 19-25 
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    ISSN: 1617-4623
    Keywords: IGF-1 ; Escherichia coli secretion/export ; LamB leader peptide ; Heterologous gene expression ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The processing of LamB-IGF-1 fusion protein and the export of processed IGF-1 (insulin-like growth-factor-1) into the growth medium was examined in the Escherichia coli host strain, JM101. Several strain or plasmid modifications were tried to increase export of periplasmic (Processed) IGF-1 into the growth medium of JM101. These included: (1) use of a lon null mutant strain to increase accumulation levels of unprocessed LamB-IGF-1 fusion protein; (2) use of an alternative drug resistance marker on the expression plasmid rather than beta-lactamase, thereby reducing any competition for processing of LamB-IGF-1 by signal peptidase; (3) examination of whether phage M13 gene III protein expression caused more periplasmic IGF-1 to be exported into the growth medium due to increased outer membrane permeability; and (4) examination of the effect of E. coli or yeast optimized IGF-1 codons. None of these strain or plasmid modifications caused any significant increase in export of IGF-1 into the growth medium of JM101. Solubility studies of LamB-IGF-1 and processed IGF-1 showed that virtually all of the LamB-IGF-1 and IGF-1 remaining within the cell after a 2 h induction period was insoluble. This implied that only soluble LamB-IGF-1 was processed to IGF-1 and that only soluble IGF-1 was exported into the growth medium. Taken together, the results indicated that LamB-IGF-1 and IGF-1 solubility were the limiting factors in secretion of IGF-1 into the periplasm and export of IGF-1 into the growth medium.
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    Characterization of genetic transformation in Streptococcus oralis NCTC 11427: Expression of the pneumococcal amidase in S. oralis using a new shuttle vector (1988)
    Springer
    Molecular genetics and genomics 215 (1988), S. 53-57 
    Details
    ISSN: 1617-4623
    Keywords: Competence ; Autolysins ; Choline ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have worked out conditions for the study of competence development and genetic transformation in Streptococcus oralis NCTC 11427 (type strain), a species that contains choline in the cell wall. The peak of competence was found at the early exponential phase of growth and the optimal conditions for transformation were achieved with shuttle plasmids prepared from S. pneumoniae or from Escherichia coli serving as donor DNA. Transformation with dye-bouyant density gradient purified plasmid preparations followed first-order kinetics. The pneumococcal amidase can be expressed in S. oralis harbouring a plasmid carrying the lytA gene. This enzyme lysed the cell wall of the transformed cell in the presence of detergents.
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    URL: http://dx.doi.org/10.1007/BF00331302
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    DNA sequence of the metC gene and its flanking regions from Salmonella typhimurium LT2 and homology with the corresponding sequence of Escherichia coli (1989)
    Springer
    Molecular genetics and genomics 216 (1989), S. 164-169 
    Details
    ISSN: 1617-4623
    Keywords: metC ; Cystathionine-β-lyase ; Nucleotide sequence ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DNA sequence of the Salmonella typhimurium metC gene and its flanking regions was determined. The metC gene contains an open reading frame of 1185 nucleotides encoding a polypeptide of 395 amino acids with a predicted molecular weight of 42874 daltons. S1 nuclease mapping experiments located the transcription start site of the metC gene. The nucleotide sequence and the deduced amino acid sequence for the metC genes of S. typhimurium and Escherichia coli were compared. Although there are 279 nucleotide replacements, most do not change the amino acid sequence. Nucleotide sequence analysis of the flanking regions of the S. typhimurium metC gene shows that there is an open reading frame upstream and an open reading frame downstream of the gene. The existence of the divergently transcribed upstream open reading frame (designated ORF1) was confirmed by the construction of an ORF1-lacZ fusion. The transcription start site of ORF1 was determined by S1 nuclease mapping.
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    URL: http://dx.doi.org/10.1007/BF00332246
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    In vivo cloning of DNA into multicopy cosmids by mini-Mu-cosduction (1986)
    Springer
    Molecular genetics and genomics 205 (1986), S. 546-549 
    Details
    ISSN: 1617-4623
    Keywords: Transposition ; Genomic libraries ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A general in vivo procedure for cloning Escherichia coli genes into cosmids has been developed. The method we describe here uses a deleted Mu phage (a mini-Mu) to transpose E. coli genes into cosmids during mini-Mu replication. The resulting cosmids clones are packaged in-vivo into λ phage particles. Plasmids carrying a particular DNA sequence can be selectively recovered after infection of a new host with the in vivo constructed genomic cosmid library. This system was used succesfully to clone several E. coli genes.
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    Cloning of two type II methylase genes that recognise asymmetric nucleotide sequences: FokI and HgaI (1987)
    Springer
    Molecular genetics and genomics 209 (1987), S. 570-574 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Methylase ; Plasmid vector ; Gel electrophoresis ; Clearage map
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The modification genes of Flavobacterium okeanokoites and Haemophilus galinarum have been cloned into the vector pBR322 and expressed in Escherichia coli cells. FokI methylase gene is contained on a 3.80 kb piece of F. okeanokoites DNA. Plasmid constructs carrying this fragment of DNA are resistant to digestion by FokI restriction endonuclease but are sensitive to cleavage by HindIII, EcoRI and PstI. Unmodified λ DNA molecules, exposed in vitro to cell extracts prepared from cells habouring this plasmid, became resistant to digestion by FokI. The smallest HgaI methylase clone carries the pBR322 plasmid containing a 3.50 kb piece of H. galinarum DNA. This plasmid is resistant to digestion by HgaI. Neither the FokI nor the HgaI restriction endonuclease was detected in either clone. This is the first report of cloning modification genes whose protein products recognise asymmetric nucleotide sequences.
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    Cloning and nucleotide sequence of the Salmonella typhimurium LT2 metF gene and its homology with the corresponding sequence of Escherichia coli (1988)
    Springer
    Molecular genetics and genomics 212 (1988), S. 246-251 
    Details
    ISSN: 1617-4623
    Keywords: metF ; 5,10-methylenetetrahydrofolate reductase ; Recombinant DNA ; Nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Salmonella typhimurium LT2 metF gene, encoding 5,10-methylenetetrahydrofolate reductase, has been cloned. Strains with multicopy plasmids carrying the metF gene overproduce the enzyme 44-fold. The nucleotide sequence of the metF gene was determined, and an open reading frame of 888 nucleotides was identified. The polypeptide deduced from the DNA sequence contains 296 amino acids and has a molecular weight of 33 135 daltons. Mung bean nuclease mapping experiments located the transcription start point and possible transcription termination region for the gene. There is a 25bp nucleotide sequence between the translation termination site and the possible transcription termination region. This region possesses a GC-rich sequence that could form a stable stem and loop structure once transcribed (ΔG=-9 kcal/mol), followed by an AT-rich sequence, both of which are characteristic of rho-independent transcription terminators. The nucleotide and deduced amino acid sequences of the S. typhimurium metF gene are compared with the corresponding sequences of the Escherichia coli metF gene. The nucleotide sequences show 85% homology. Most of the nucleotide differences found do not alter the amino acid sequences, which show 95% homology. The results also show that a change has occurred in the metF region of the S. typhimurium chromosome as compared to the E. coli chromosome.
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    The repressor gene (c) of the Streptomyces temperate phage ϕc31: Nucleotide sequence, analysis and functional cloning (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 269-277 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Helix-turn-helix motifs ; In vitro transcription-translation ; Phage immunity ; Exonuclease III deletions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleotide sequence of the 3.4 kb SphI-G fragment that contained the repressor gene (c) of the temperate Streptomyces phage ϕc31 was determined. Analysis of this sequence revealed a large open reading frame with protein coding character and sequence changes in c gene point and deletion mutants identified this as the coding region of the repressor. Two of the mutants studied had undergone deletions of 1.1 kb and 1.4 kb that had occurred across short direct repeats of 6 bp and 11 bp, respectively. Coupled in vitro transcription-translation experiments using the cloned SphI-G fragment and Streptomyces lividans cell free extracts identified a protein product of approximately 72 kDa, in close agreement with that predicted from the nucleotide sequence. A strongly predicted helix-turn-helix motif that may be involved in DNA binding occurred towards the carboxy-terminus of the amino acid sequence. Initial attempts to clone the SphI-G fragment in Streptomyces failed; using information gained from the sequence analysis a smaller segment of this DNA fragment was cloned in S. lividans and conferred immunity to a clear plaque mutant (c1) of ϕc31.
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    URL: http://dx.doi.org/10.1007/BF00339591
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    Cloning of two chloramphenicol acetyltransferase genes from Clostridium butyricum and their expression in Escherichia coli and Bacillus subtilis (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 328-332 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Chloramphenicol resistance ; cat gene ; Plasmids pC194 and pUB110 ; Inducible gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two non-homologous chloramphenicol (Cm) acetyltransferase (CAT) genes, designated catA and catB, were cloned from Clostridium butyricum type strains and characterized by restriction mapping. Both genes are efficiently expressed in Escherichia coli and Bacillus subtilis. In contrast to analogous genes from staphylococci and bacilli, gene expression is not dependent on induction by Cm. The genes are considered as chromosomal, since no association with endogenous plasmids was detectable. Southern hybridization revealed a homology between catA and the staphylococcal Cm resistance plasmid, pC194. The subunit size of the clostridial CAT enzymes expressed in E. coli was determined as 22.5 kDa (catA) and 24 kDa (catB), respectively. The C. butyricum cat genes provide potentially useful selection markers for the construction of cloning vectors from cryptic clostridial plasmids.
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  • 47
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    Comparison of injection-generated gradient and isocratic ion-pair chromatography on short microbore column in HPLC (1986)
    Springer
    Chromatographia 22 (1986), S. 311-315 
    Details
    ISSN: 1612-1112
    Keywords: HPLC ; Ion-pair chromatography ; Short microbore column ; Catecholamine analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Possibilities of using short microbore columns in trace analysis are presented. Retention of ionizing solutes (catecholamines) is compared using both isocratic ionpair chromatography and injection-generated gradient of a counter ion in the mobile phase. Column efficiency is maintained even with a relatively high sample volume.
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    Chromatographic and NMR study of high boiling oils (1985)
    Springer
    Chromatographia 20 (1985), S. 609-614 
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    ISSN: 1612-1112
    Keywords: petroleum analysis ; Hydrocarbon group determination ; HPLC ; NMR ; Adsorption chromatography of oils
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary This paper presents an analytical system for the rapid and effective analytical control of hydrocarbon oil fractions. Three instrumental analytical methods were used: (1) semipreparative liquid adsorption chromatography (LC) (2) high-performance liquid chromatography (HPLC) (3) nuclear magnetic resonance spectroscopy (NMR). These methods were successfully applied for the determination of aliphatic, naphthenic and aromatic compounds in oils used in the preparation of motor oils. For low aromatic content the concentration of naphthenic and aliphatic ompounds may be determined by analyzing the original oil samples. In other cases the fractions obtained by separation on the semipreparative column were also analyzed. The combination of LC, HPLC and NMR gives a good possibility for the identification and determination of hydrocarbon groups in distillation oil samples.
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    Simultaneous high-performance liquid chromatographic determination of phenazopyridine and nitrofurantoin in tablets (1988)
    Springer
    Chromatographia 25 (1988), S. 61-63 
    Details
    ISSN: 1612-1112
    Keywords: Phenazopyridine ; Nitrofurantoin ; HPLC ; Tablets
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A reversed-phase, High-performance Liquid chromatographic method for the simultaneous determination of phenazopyridine and nitrofurantoin in tablets is described. An aminopropyl-silica (APS-Hypensil) 5μm column and a mobile phase consisting of Methanol:H2O: 0.05M sodium dihydrogen phosphate (50∶45∶5) were used. Calibration curves were linear over the concentration range 2–20μg/ml, with minimum detectability of 10ng/ml for both drugs. The method was applied to tablets containing the two species and the results obtained were compared to those given with a published method.
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    Distribution and characterization of neuropeptide Y-like immunoreactivity in the brain and pituitary of the goldfish (1989)
    Springer
    Cell & tissue research 255 (1989), S. 529-538 
    Details
    ISSN: 1432-0878
    Keywords: Neuropeptide ; Pituitary ; Brain ; Immunocy tochemistry ; HPLC ; Carassius auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of neuropeptide Y (NPY) immunoreactivity has been studied by means of immunocytochemistry and radioimmunoassay in the brain of the goldfish. It was found that NPY had a widespread distribution in the entire brain in particular in the telencephalon, diencephalon, optic tectum and rhombencephalon. In the pituitary gland, positive type-B fibers were observed in the various lobes frequently in direct contact with secretory cells, in particular the gonadotrophs, somatotrophs and MSH (melanocyte-stimulating hormone) secreting cells. When measured by radioimmunoassay, the highest NPY concentrations were found in the pituitary and telencephalon, confirming the results of immunocytochemistry. The displacement curves obtained with serial dilutions of brain extracts were parallel to that of synthetic porcine NPY. Following high performance liquid chromatography, the NPY-like material extracted from goldfish brain co-eluted as a single peak with synthetic porcine NPY. These data demonstrate the presence of an NPY-like substance widely distributed in the goldfish brain. The observation of NPY-immunoreactive fibers in the pituitary gland suggests that, among its other functions, NPY may play a role in the neuroendocrine regulation of pituitary function.
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    Disposition of 2′, 3′-dideoxyadenosine and 2′, 3′-dideoxyinosine in mice (1989)
    Springer
    Investigational new drugs 7 (1989), S. 139-146 
    Details
    ISSN: 1573-0646
    Keywords: 2′, 3′-dideoxyadenosine ; 2′-deoxycoformycin, 2′ ; 3′-dideoxyinosine ; metabolites ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary Mice were dosed with [3H]2′,3′-dideoxyadenosine ([3H]ddA) in three procedures: intravenously, intraperitoneally, and interperitoneally following a dose of 2′ -deoxycoformycin (dCF). For mice dosed intravenously, the content of radioactivity in plasma and tissue samples were essentially constant after 30 min. Of the radioactivity in plasma and brain samples collected between 30 min and 24 hr, more than 94% was present as 3H2O, indicating that most of the tritium from [3H]ddA had exchanged with water. No intact ddA was detected, and the deamination product, 2′,3′ -dideoxyinosine (ddI), was present only transiently. In the urine, the major radioactive material was [3H]ddI. Also detected were 3H2O and small amounts of [3H]hypoxanthine and [3H]ddA. Following intraperitoneal doses to mice, levels of radioactivity in plasma, liver, and kidney increased to a maximum by 15–30 min after dosing but dropped to essentially constant levels thereafter, again indicating that the tritium had exchanged with water. At 5, 15, and 30 min after dosing, ddI was the major radioactive component in plasma. Only small amounts of ddA were present. When dCF was administered 24 hr prior to intraperitoneal [3H]ddA, levels of radioactivity in plasma, liver, and kidney reached a maximum at 30 to 60 min after dosing and decreased to essentially constant levels thereafter. The dCF transiently inhibited the deamination of ddA to ddI, since, in plasma, [3H]ddA was the main radioactive component at 5 and 15 min after dosing. Comparison of HPLC assays based on radioactivity detection and UV absorbance showed that they were equivalent for measuring ddA and ddI in samples derived from dosed mice. Therefore, exchange of tritium must have occurred at a metabolic step beyond ddI. For mice dosed intravenously and orally with unlabeled ddI, there was evidence of a saturated process. Nevertheless, for the high and low intravenous doses of ddI, the percent of dose excreted in the urine as unchanged drug was the same.
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    Chromatographic separation of asparagine-linked oligosaccharides labeled with an ultravioletabsorbing compound,p-aminobenzoic acid ethyl ester (1988)
    Springer
    Glycoconjugate journal 5 (1988), S. 13-26 
    Details
    ISSN: 1573-4986
    Keywords: asparagine-linked oligosaccharides ; UV-absorbing derivatives ; gel permeation chromatography ; HPLC ; p-aminobenzoic acid ethyl ester
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract We have expanded on the suitability ofp-aminobenzoic acid ethyl ester as an ultraviolet-absorbing reagent [Wanget al., (1984) Anal Biochem 141:366–81] for the analysis of asparagine-linked oligosaccharides derived from glycoproteins. The oligosaccharides released from glycoproteins by hydrazinolysis/N-reacetylation were derivatized withp-aminobenzoic acid ethyl ester and the derivatives were purified and separated into neutral and acidic oligosaccharides on a PRE-SEP C18 cartridge. The acidic oligosaccharides could be further separated into a few species by high-voltage paper electrophoresis. p-Aminobenzoic acid ethyl ester derivatives of neutral oligosaccharides were analyzed by gel permeation chromatography on Bio-Gel P-4 and HPLC on a silica-based amide column. The elution profile and the proportion of the oligosaccharides were in agreement with literature values. The overall yield of oligosaccharides from glycoproteins was approximately 70%. Fifty pmol of oligosaccharide were detectable on Bio-Gel P-4 and 4–5 pmol on HPLC.
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    Theβ-subunit of human chorionic gonadotropin containsN-glycosidic trisialo tri- and tri′-antennary carbohydrate chains (1988)
    Springer
    Glycoconjugate journal 5 (1988), S. 221-233 
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    ISSN: 1573-4986
    Keywords: hCG ; PNGase-F ; FPLC ; HPLC ; 1 H-NMR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract TheN-linked carbohydrate chains of theβ-subunit of highly purified urinary human chorionic gonadotropin have been re-investigated. The oligosaccharides were released enzymatically by peptide-N 4-(N-acetyl-β-glucosaminyl)asparagine amidase-F, and fractionated by a combination of FPLC and HPLC. As a result of the application of improved fractionation methods, apart from the earlier reported carbohydrate chains, also small amounts of trisialo tri- and tri′-antennary oligosaccharides were found. The primary structures of the latter carbohydrate chains have been determined by 500-MHz1H-NMR spectroscopy to be
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    Reverse phase HPLC fractionation of the oligosaccharide alditols isolated from an I-active ovarian cyst mucin glycoprotein (1985)
    Springer
    Glycoconjugate journal 2 (1985), S. 17-30 
    Details
    ISSN: 1573-4986
    Keywords: HPLC ; cyst mucin glycoprotein ; blood group I
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract We have used reverse phase high pressure liquid chromatography (HPLC) to separate the reduced oligosaccharides produced by alkaline borohydride degradation of ovarian cyst blood group substances. From a single cyst, six oligosaccharides, ranging from two to seven residues in length, have been isolated by preparative HPLC on C-18 stationary phases using water for elution. The purity of the products and their structures were determined by high field proton NMR spectroscopy in conjunction with exo-and endoglycosidase digestion. All the chains isolated terminated inN-acetylgalactosaminitol which was substituted at the 3-postion by galactose and in some cases at the 6-postion byN-acetylglucosamine. The largest identified oligosaccharide was a heptasaccharide alditol containing a single α-linked fucose in a Lewis blood group structure (Lea).
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    Retention behaviour of some coumarins in normal-phase high-performance thin-layer and column chromatography (1988)
    Springer
    Chromatographia 25 (1988), S. 111-116 
    Details
    ISSN: 1612-1112
    Keywords: Coumarins ; HPTLC ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A number of coumarins, furocoumarins and pyranocoumarins were investigated using HPTLC and HPLC systems consisting of silica gel and binary and ternary solvents containing a polar modifier (acetonitrile, dioxane, diisopropyl ether, methyl ethyl ketone, ethyl acetate, 2-propanol or tetrahydrofuran) and a non-polar or weakly polar diluent (n-heptane or dichloromethane). The experimental results obtained on thin layers, under isocratic conditions, showed a linear relationship between the RM values and the log of the concentration of the polar modifier. The influence of the modifier and the individual substituents in the solute molecule on retention is presented as chromatographic “spectra” by plotting the RM and logk′ values against the mobile phase used.
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    Characterisation of neuropeptides of the AKH/RPCH-family from corpora cardiaca of Coleoptera (1989)
    Springer
    Journal of comparative physiology 159 (1989), S. 589-596 
    Details
    ISSN: 1432-136X
    Keywords: Corpora cardiaca ; Coleoptera ; Neuropeptides ; HPLC ; Amino acid composition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Extracts of corpora cardiaca from two members of the family Tenebrionidae,Zophobas rugipes andTenebrio molitor, from one member of the Chrysomelidae,Leptinotarsa decemlineata, and from three members of the Scarabaeidae,Pachnoda marginata, P. sinuata andMelolontha hippocastani, were assayed for adipokinetic and hypertrehalosaemic activity in acceptor locusts (Locusta migratoria) and cockroaches (Periplaneta americana), respectively. All corpus cardiacum material tested, except that from the cockchafer,M. hippocastani, gave positive bioassay results. Biological activities of corpus cardiacum extracts from all species investigated can be resolved on reversed-phase high performance liquid chromatography (RP-HPLC). Gland extracts from the two tenebrionid species each show a single peak of biological activity associated with a single peak of UV absorbance having an identical retention time in both species. The two biologically active fractions from the corpora cardiaca of the potato beetle,L. decemlineata, coelute with exogenous (synthetic) hypertrehalosaemic hormones I and II of the American cockroach. The two species of the genusPachnoda contain two active compounds in their glands; compound I of each species is more abundant and elutes just ahead of the (synthetic) hypertrehalosaemic hormone of the cockroachBlaberus discoidalis. The gland material ofM. hippocastani exhibits and absorbance peak with the same retention time as the major peak from thePachnoda-species; however, this peak material does not elicit biological activity in the assays used here. After fractionation by RP-HPLC the main biologically active compounds were subjected to amino acid analyses. All factors are peptidic and contain 8 amino acid residues. The peptides from the tenebrionid species have the amino acid residues Asx(2), Glx(1), Ser(1), Pro(1), Leu(1), Phe(1) and Trp(i), whereas the main peptide from corpora cardiaca ofP. marginata contains the residues Asx(2), Glx(1), Ser(1), Pro(1), Tyr(1), Leu(1) and Trp(1). Amino acid composition analyses of the two active fractions fromL. decemlineata reveal the residues Asx(2), Glx(1), Ser(1), Pro(1), Val(1), Phe(1) and Trp(1) for compound I and Asx(1), Glx(1), Thr(2), Pro(1), Leu(1), Phe(1) and Trp(1) for compound II.
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    Dopamine in tissues of the hydrozoan jellyfishPolyorchis penicillatus as revealed by HPLC and GC/MS (1989)
    Springer
    Journal of comparative physiology 159 (1989), S. 173-181 
    Details
    ISSN: 1432-136X
    Keywords: Jellyfish ; Dopamine ; HPLC ; GC/MS ; Hydrozoa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary High performance liquid chromatography of alumina extracts of several tissues inPolyorchis penicillatus show the presence of dopamine and a catecholamine resembling norepinephrine. Dopamine is found in the highest concentrations in nerve-rich tissue (120 fmol·mg wet wt−1), at intermediate concentrations in endoderm-rich tissue (30 fmol·mg−1), and at the lowest concentrations in the mesoglea (10 fmol·mg−1). The presence of dopamine was confirmed using gas chromatography/mass spectrometry with negative ion chemical ionization, but norepinephrine and epinephrine could not be detected in nerve-rich tissue.
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    Luminol chemiluminescence HPLC reaction detector for amino acids and other ligands (1987)
    Springer
    Microchimica acta 92 (1987), S. 79-90 
    Details
    ISSN: 1436-5073
    Keywords: luminol chemiluminescence ; HPLC ; reaction detector ; amino acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract A detector for HPLC is based on suppression of chemiluminescence from the Cu(II)-luminol-peroxide reaction. Analytes which complex Cu(II) reduce the free Cu(II) concentration and thus the observed chemiluminescence intensity. The degree of chemiluminescence suppression is a measure of the analyte concentrations. Detection limits are in the range of 1 pmole-1 nmole for amino acids (both primary and secondary without derivatization), CN−, amines, catecholamines, catechol, and aminoglycoside antibiotics. The detection approach is demonstrated with an ion-exchange separation of amino acids, a reverse phase separation of catecholamines, and an ion-pair separation of the three components of gentamicin C in commercial gentamicin sulfate.
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    The continuous infrared spectroscopic analysis of reversed phase liquid chromatography separations (1988)
    Springer
    Microchimica acta 95 (1988), S. 185-187 
    Details
    ISSN: 1436-5073
    Keywords: infrared ; FTIR ; HPLC ; reflectance-absorbance ; HPLC/IR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract A method for HPLC/FTIR is discussed in which the effluent from a microbore HPLC is continuously deposited onto and eliminated from the surface of a circular rotating germanium crystal that has been aluminum coated on its opposite side. After deposition, the germanium disk is again rotated, this time in the sample compartment of an FTIR spectrometer while reflectance-absorbance infrared spectra are continuously collected. The novel germanium-aluminum deposition surface allows collection of reflectance-absorbance spectra that are free of the degrading effects of superposition phenomena characteristic of reflectance-absorbance spectra obtained on metal surfaces. Furthermore, germanium is impervious to aqueous solvent mixtures and, therefore, allows for the direct deposition of reversed phase separations, including those requiring acid modified mobile phases.
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    Method for determining airborne diisocyanates (1988)
    Springer
    Microchimica acta 96 (1988), S. 197-206 
    Details
    ISSN: 1436-5073
    Keywords: diisocyanates (2,4-TDI, HDI, MDI) ; chemisorption ; HPLC ; air
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract A method has been developed for quantitatively determining 3 typical diisocyanates, viz. 2,4-toluylene diisocyanate (2,4-TDI), hexamethylene diisocyanate (HDI) and 4,4′-diphenyl-methane diisocyanate (MDI). The technique relies on chemisorption onto a suitable carrier material contained in an absorption tube coated with 1-(2-pyridyl)piperazine (2-PP). Stable urea derivatives with high molar absorptivity are formed, desorbed from the carrier material, separated by means of highpressure liquid chromatography and identified. The limits of detection for a sample volume of 20 1 of air are 0.9μg m−3 for 2,4-TDI and HDI, and 0.6μg m−3 for MDI. The derivatives are stable at 4°C for longer than 30 days. The influence of atmospheric humidity and the interference of dibutylamine on the detection reaction were investigated for 2,4-TDI and HDI. The whole procedure is easy to perform, highly sensitive and very reproducible and is suitable for monitoring concentrations of 2,4-TDI, HDI and MDI.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01236104
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  • 61
    Electronic Resource
    Electronic Resource
    Cloning and sequence analysis of the 18 S ribosomal RNA gene of tomato and a secondary structure model for the 18 S rRNA of angiosperms (1989)
    Springer
    Molecular genetics and genomics 219 (1989), S. 17-25 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Small subunit ; Ribosomal DNA ; Sequence comparison ; Lycopersicon esculentum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The gene of a cytoplasmic 18 S ribosomal RNA (18 S rDNA) of the dicotyledonous plant tomato (ycopersicon esculentum) cv. Rentita has been cloned, and its complete primary structure has been determined. The tomato 18 S rDNA is 1805 by long with a G+C content of 49.6%. Its sequence exhibits 94%–96% positional identity when it is colinearly aligned with the previously reported sequences of the 17–18 S rDNAs of the dicot soybean and the monocots maize and rice. A model of the secondary structure of the 18 S rRNA of angiosperms is presented and its genera-specific structural features are compared with a current eukaryotic 18 S rRNA consensus model.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00261152
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