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  • Radioactivity  (1,487)
  • Photosystem II  (1,079)
  • HPLC  (1,020)
  • (Human)
  • Gramineae
  • Solanaceae
  • recombinant DNA
  • Elsevier  (6,448)
  • Springer  (730)
  • Coimbra University Press  (1)
  • Lagos (Nigeria)  (1)
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  • 1
    facet.materialart.
    Unknown
    Biofísica médica: exercícios práticos (2021)
    Coimbra University Press
    Details
    Publication Date: 2022-01-31
    Description: The Book of Medical Biophysics. Practical exercises intends to be an auxiliary for the study of this matter. It constitutes the compilation of the exercises developed by the set of assistants since the Faculty of Medicine of the University of Coimbra has decided to invest in specific training in biophysics. The issues for which problems have been made relate to 3 chapters: mass transport through membranes, fluids and radioactivity. All problems developed are aimed at biomedical applications, and assist in the practical comprehension of the same themes presented in the book Biophysics Medical, authored by João José Pedroso de Lima, also published by the Coimbra University Press and representing the recommended book for the discipline. In fact, the theoretical explanations are even referred to this book. In all chapters there are problems resolved and commented, other problems for which solutions are present and yet another type that are proposed problems, but no solution presented. We hope that this book will be a good helper for the study of biophysics.
    Keywords: Medicine ; Medical biophysicsm - education ; Membranes ; Fluids ; Radioactivity
    Language: Portuguese
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    E-BOOK
  • 2
    facet.materialart.
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    Gross alpha and beta surveys of Lake Kainji water (2021)
    FISON | Lagos (Nigeria)
    In:  http://aquaticcommons.org/id/eprint/24608 | 19325 | 2018-05-22 05:54:40 | 24608 | Fisheries Society of Nigeria
    Details
    Publication Date: 2021-07-15
    Description: A gross alpha and beta survey of the water of Kainji Lake was conducted using multi-channel analyzer, a proportional counter detector. This was done to estimate the gross radioactivity concentrations due to natural radionuclides in the lake water. Radioactivity, a phenomenon that leads to production of radiations, and radiation is known to trigger or induce cancer. The gross alpha and beta activity concentration analyzed in the water shows that the alpha activity ranges from 1.0 x 105 Bq/cm3 to 0.7Bq/cm3 with an average value indication of 0.0702Bq/cm3, whereas the beta activity concentration indicated BDL for all the sites with exception of upstream 01 location (9~' 51' 485' and 4~'35' 473'E) which has 0.0022Bq/cm3. This could be attributed to the continued flow of the lake water. Although radioactivity measured in lakes is usual higher than that of flowing waters, because a significant part of the radioactive substance brought in by other tributary rivers accumulates in it (Szaho et al.1998). That of Kainji Lake is entirely different.
    Description: Includes: 9 references.
    Keywords: Limnology ; Pollution ; Nigeria ; Kainji L. ; Gross alpha-beta decay ; Radioactivity ; Lake Kainji ; Contamination ; freshwater environment ; automation
    Repository Name: AquaDocs
    Type: conference_item , TRUE
    Format: application/pdf
    Format: application/pdf
    Format: 164-166
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    PAPER (OA)
  • 3
    Electronic Resource
    Electronic Resource
    Cytokinins in pathogenesis and disease resistance of Pyrenophora teres-barley and Dreschslera maydis-maize interactions during early stages of infection (2000)
    Springer
    Mycopathologia 148 (2000), S. 87-95 
    Details
    ISSN: 1573-0832
    Keywords: Autoradiography ; barley ; cytokinins ; Dreschslera maydis ; green islands ; HPLC ; maize ; Pyrenophora teres
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Infection of Hordeum vulgare L. by Pyrenophora teresand of Zea mays by Dreschslera maydis were characterized by ‘green island’ formation, higher cytokinin levels and accumulation of metabolites in the infected areas. Higher cytokinin concentrations of the order 6-Y,Y-dimethylallylaminopurine 〉 zeatinriboside 〉 zeatin 〉dihydrozeatinriboside were detected at infection sites of susceptible hosts. By virtue of these cytokinins, infection sites may be acting as metabolic sinks helping proliferation of the pathogen. Existence of translocatory sinks at infection zones was confirmed from autoradiographic studies,where, accumulation of labeled metabolites was prominent at infection sites of susceptible hosts. Upon infection the lower cytokinin levels of resistant hosts decreased further with progress of infection. In the infected resistant hosts the concentrations of zeatin/zeatinriboside were the maximum among the four identified cytokinins. The pathogen is also capable of secreting cytokinins as evident from quantification of cytokinins in culture filtrate extracts using HPLC. Since detached leaves were used in the experiments the increase/decrease of various cytokinin levels may be attributed to pathogen influence. The increase in cytokinin levels in the susceptible host may be aiding the growth of the pathogen on one hand, while the decrease in the infected resistant host may signal the host to activate defenses against a potential pathogen at the early stage of infection.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007126025955
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  • 4
    Electronic Resource
    Electronic Resource
    Genomic factors responsible for abnormal morphology of pollen tubes in the interspecific cross Nicotiana tabacum ×N. rustica (2000)
    Springer
    Sexual plant reproduction 12 (2000), S. 333-337 
    Details
    ISSN: 1432-2145
    Keywords: Key words Unilateral incongruity ; Interspecific cross ; Gemome ; Solanaceae ; Tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Nicotiana tabacum shows unilateral pollen-pistil incongruity with N. rustica. If N. tabacum is pollinated with N. rustica, growth of the pollen tube is arrested in the middle of the style, and abundant callose deposition, tube swelling and tube winding are observed. An attempt was made to clarify the genomic factors responsible for this pollen-pistil incongruity. N. tabacum was pollinated with N. paniculata or N. undulata, progenitors of amphidiploid N. rustica. When pollinated with N. undulata, growth of the pollen tube was arrested in the middle of the style and showed abnormal morphology similar to that with N. rustica, but when pollinated with N. paniculata the pollen tube reached near the base of the style and was almost normal in appearance. These observations suggest that the factors responsible for the pollen tube abnormality of N. rustica are derived from the N. undulata genome.We also used N. sylvestris, N. glutinosa and N. otophora as pistilar parents and N. rustica or its progenitors as pollen parents to examine the genomic factors of the pistilate parents. The pollen tube features of these three species in the pistils of N. sylvestris were similar to those in the pistil of N. tabacum.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004970000027
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  • 5
    Electronic Resource
    Electronic Resource
    A two-dimensional many-body system with competing interactions as a model for segregation of photosystems in thylakoids of green plants (2000)
    Springer
    European biophysics journal 29 (2000), S. 214-220 
    Details
    ISSN: 1432-1017
    Keywords: Key words Lipid-protein interactions ; Photosystem I ; Photosystem II ; Thylakoid membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract  We address the segregation of photosystems I (PSI) and II (PSII) in thylakoid membranes by means of a molecular dynamics method. We assume a two-dimensional (in-plane) problem with PSI and PSII being represented by particles with different values of negative charge. The pair interactions between particles include a screened Coulomb repulsive part and am exponentially decaying attractive part. Our modeling results suggest that the system may have a complicated phase behavior, including a quasi-crystalline phase at low ionic screening, a disordered phase and, in addition, a possible “clotting” agglomerate phase at high screening where the photosystems tend to clot together. The relevance of the observed phenomena to the stacking of thylakoid membranes is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002490000080
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  • 6
    Electronic Resource
    Electronic Resource
    Interaction of nitric oxide with the oxygen evolving complex of photosystem II and manganese catalase: a comparative study (2000)
    Springer
    JBIC 5 (2000), S. 354-363 
    Details
    ISSN: 1432-1327
    Keywords: Catalase ; Manganese cluster ; Nitric oxide ; Photosystem II
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Thermus thermophilus catalase. Flash fluorescence studies indicate that the S3 state of the OEC in the presence of ca. 0.6 mM NO is reduced to the S1 with an apparent halftime of ca. 0.4 s at about 18 °C, compared with a biphasic decay, with approximate halftimes of 28 s for S3 to S2 and 140 s for S2 to S1 in the absence of NO. Under similar conditions the S2 state is reduced by NO to the S1 state with an approximate halftime of 2 s. These results extend a recent study indicating a slow reduction of the S1 state at −30°C, via the S0 and S−1 states, to a Mn(II)-Mn(III) state resembling the corresponding state in catalase. The reductive mode of action of NO is repeated with the di-Mn cluster of catalase: the Mn(III)-Mn(III) redox state is reduced to the Mn(II)-Mn(II) state via the intermediate Mn(II)-Mn(III) state. The kinetics of this reduction suggest a decreasing reduction potential with decreasing oxidation state, similar to what is observed with the active states of the OEC. What is unique about the OEC is the rapid interaction of NO with the S3 state of the OEC, which is compatible with a metalloradical character of this state.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/PL00010664
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  • 7
    Electronic Resource
    Electronic Resource
    Microsatellites and microsynteny in the chloroplast genomes of Oryza and eight other Gramineae species (2000)
    Springer
    Theoretical and applied genetics 100 (2000), S. 1257-1266 
    Details
    ISSN: 1432-2242
    Keywords: Key words Chloroplast microsatellite ; Simple sequence length polymorphism (SSLP) ; Allelic diversity ; Rice (Oryza sativa) ; Gramineae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Primer pairs flanking ten chloroplast microsatellite loci, originally identified in Oryza sativa cv Nipponbare, were evaluated for amplification and allelic diversity using a panel of 13 diverse cultivars of rice (O. sativa), 19 accessions of wild rice (three O. officinalis, five O. latifolia, five O. minuta, four O. australiensis, one O. brachyantha and one O. ridleyi) and eight other Gramineae species (maize, teosinte, wheat, oat, barley, pearl millet, sorghum and sugarcane). Amplified products were obtained for all samples at nine out of ten loci. Among the rice cultivars, the number of alleles per locus ranged from one to four, with monomorphic patterns observed at five loci. The average polymorphism information content (PIC) value at the other five (polymorphic) loci was 0.54 among the 13 cultivars. When wild rice and the other Gramineae species were compared based on the proportion of shared alleles, their phylogenetic relationships were in agreement with previous studies using different types of markers; however, the magnitude of the differences based on chloroplast microsatellites underestimated the genetic distance separating these divergent species and genera. A sequence-based comparison of homologous regions of the rice and maize chloroplast genomes revealed that, while a high level of microsynteny is evident, the occurrence of actively evolving microsatellite motifs in specific regions of the rice chloroplast genome appears to be mainly a species or genome-specific phenomenon. Thus the chloroplast primer pairs used in this study bracketed mutationally active microsatellite motifs in rice but degenerate, interrupted motifs or highly conserved, mutationally inert motifs in distantly related genera.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220051432
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  • 8
    Electronic Resource
    Electronic Resource
    Molecular mapping of chromosome segments introgressed from Solanum lycopersicoides into cultivated tomato (Lycopersicon esculentum) (2000)
    Springer
    Theoretical and applied genetics 100 (2000), S. 232-241 
    Details
    ISSN: 1432-2242
    Keywords: Key words Comparative mapping ; Homoeologous recombination ; Segregation distortion ; Solanaceae ; Tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The wild nightshade Solanum lycopersicoides (accessionLA2951) was backcrossed to the cultivated tomato (Lycopersicon esculentum cv ’VF36’), then inbred through single-seed descent for several generations. Over 300 backcross-inbred families thereby derived were genotyped at 139 marker loci, consisting of RFLPs, allozymes, and monogenic morphological markers, to identify introgressed S. lycopersicoides chromosomes and segments thereof. The pattern of genotypes observed in the lines indicated a high degree of overall synteny between the S. lycopersicoides genome and that of tomato. Two putative single-copy RFLP probes revealed secondary loci in this wide cross. Recovery of the L. esculentum genome was more rapid than expected, with an average value in the BC2 generation of 97.8%, versus the expected value of 87.5%. This was due to widespread segregation distortion that favored L. esculentum alleles as well as a tendency for plants homozygous for in- trogressed segments to be partially or completely male-sterile, thereby preventing the fixation of S. lycopersicoides markers in many lines. Despite these difficulties, nearly every S. lycopersicoides marker (or approximately 98% of the genome, measured in centi Morgans) was represented in at least 1 backcross-inbred line, with only a region on chromosome 4L missing from the population as a whole. Although the extent of transmission and fixation of introgressed segments varied according to chromosome, overall approximately 66% of the S. lycopersicoides genome was represented by homozygous in- trogressions with sufficient fertility to reproduce by self-pollination. An excess of terminal (vs. interstitial) segments was noted, and putative heterozygous substitutions for chromosomes 6, 7, 8, and 10 were found. Recombination within certain introgressed regions was reduced over 100-fold. These backcross-inbred lines are expected to facilitate the genetic analysis of traits identified in S. lycopersicoides and their transfer into horticultural tomatoes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220050031
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  • 9
    Electronic Resource
    Electronic Resource
    Identification and comparative mapping of a dominant potyvirus resistance gene cluster in Capsicum (2000)
    Springer
    Theoretical and applied genetics 101 (2000), S. 852-859 
    Details
    ISSN: 1432-2242
    Keywords: Key words PVY ; TEV ; Lycopersicon, Solanum ; Solanaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The dominant gene Pvr7 from Capsicum chinense Jacq. ’PI159236’ confers resistance to the pepper mottle potyvirus (PepMoV) Florida (V1182) strain. This gene is tightly linked to the dominant potyvirus resistance gene Pvr4 with observed recombination frequencies of 0.012 to 0.016. A cleaved amplified polymorphic sequence (CAPS) marker linked to Pvr4 was used to localize Pvr4 and, by extension, Pvr7, to linkage group 10 on an interspecific map of pepper. Our results indicated that Pvr4, Pvr7, and Tsw, a gene conferring resistance to tomato spotted wilt virus, comprise the first identified cluster of dominant disease resistance genes in Capsicum L. This position does not correspond to the locations of dominant potyvirus resistance genes in potato or to the positions of any other mapped solanaceous resistance genes or resistance gene homologues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s001220051552
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  • 10
    Electronic Resource
    Electronic Resource
    Preparation and Chromatographic Application of Macroporous Silicate in a Capillary (2000)
    Springer
    Journal of sol gel science and technology 19 (2000), S. 371-375 
    Details
    ISSN: 1573-4846
    Keywords: phase separation ; silica ; capillary column ; HPLC ; CEC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Continuous macroporous silica gel networks were prepared in a fused silica capillary, and evaluated in reversed-phase liquid chromatography. Under pressure-driven conditions, considerable dependence of column efficiency on the linear velocity of the mobile phase was observed in spite of the small size of the silica skeletons. A major source of band broadening in the pressure-driven mode was found in the A-term of van Deemter equation. The performance of the continuous silica capillary column in the electro-driven mode was much better than that in the pressure-driven mode.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008770707572
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  • 11
    Electronic Resource
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    Extraction of pigments and fatty acids from the green alga Scenedesmus obliquus (Chlorophyceae) (2000)
    Springer
    Aquatic ecology 34 (2000), S. 119-126 
    Details
    ISSN: 1573-5125
    Keywords: carotenoids ; chlorophyll ; GC ; HPLC ; lipids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this paper, the efficiency of pigment and fatty acid extraction from resistant algae using Scenedesmus obliquus as an example was examined. We found that adding quartz sand and solvent to freeze-dried algal material and subsequent extraction in an ultrasound bath for 90min at −4 °C resulted in excellent extraction of these compounds. This extraction method was compared with a method regularly used for extraction of fatty acids and pigments, i.e. addition of solvents to algal material with subsequent incubation. Our extraction using the ultrasound and sand method was about twice as efficient as this method for both pigments and fatty acids. The ultrasound method is simple, extracts over 90% of the different substances in one step and conserves the relationships of pigments and fatty acids. In addition, no alteration- or breakdown products were observed with the new method. Thus, this method allows accurate quantitative extraction of both pigments and fatty acids from Scenedesmus obliquus and other algae. The method was also been found to be as effective for Cryptomonas erosa (Cryptophyceae), Cyclotella meneghiniana (Bacillariophyceae), Microcystis aeruginosa (Cyanophyceae), and Staurastrum paradoxum (Chlorophyceae, Desmidiaceae) and is thus applicable to a wide spectrum of algae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009911418606
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  • 12
    Electronic Resource
    Electronic Resource
    Nannochloropsis (Eustigmatophyceae) as source of commercially valuable pigments (2000)
    Springer
    Journal of applied phycology 12 (2000), S. 249-255 
    Details
    ISSN: 1573-5176
    Keywords: pigments ; ketocarotenoids ; xanthophyll cycle ; microalgae culture ; Nannochloropsis ; Eustigmatophyceae ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pigment composition and its variation with culture agewere analyzed in six strains of Nannochloropsis(Eustigmatophyceae). The capacity for accumulationof the ketocarotenoids astaxanthin and canthaxanthinwas higher in N. salina and N. gaditanathan in the other strains studied here. Theinfluence of salinity (15 to 100 practical units) onpigment production was studied in N. gaditana,where a defined pattern of variation could not befound apart from a notable increase in zeaxanthin at100‰. In cultures grown in a photobioreactor and athigh cell densities of about 109 cells mL-1,pigment production reached: 350 mg L-1 forchlorophyll a, 50 mg L-1 for violaxanthin,5 mg L-1 for canthaxanthin, 3 mg L-1 forastaxanthin. The highest contents of canthaxanthin andastaxanthin obtained in experiments with N.gaditana were 19.4 and 14.6 ng pigment (106cells)-1, respectively, which accounts for 0.7%dry weight. By means of xanthophyll cycle inductionthrough exposure of cells to high irradiance and at40 °C, conversion of violaxanthin intozeaxanthin may attain up to 70% of the violaxanthincontent, which corresponds to 0.6% dry weight. Theresults indicate that interest in Nannochloropsis as a source of valuable pigments isnot related to its capacity for single pigmentaccumulation, but the availability of a range ofpigments such as chlorophyll a, zeaxanthin,canthaxanthin and astaxanthin, each with highproduction levels.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008170915932
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  • 13
    Electronic Resource
    Electronic Resource
    Gene Transfer In Vitro and In Vivo by Cationic Lipids Is Not Significantly Affected by Levels of Supercoiling of a Reporter Plasmid (2000)
    Springer
    Pharmaceutical research 17 (2000), S. 967-973 
    Details
    ISSN: 1573-904X
    Keywords: cationic lipids ; transfection ; DNA supercoiling ; HPLC ; lipofection ; gene therapy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. It is a common preconception that supercoiledplasmid DNA is more desirable for the transfection of cells that the relaxedform of the plasmid. This notion has led to the recommendation that aspecification for the minimum amount of plasmid in the supercoiled formshould exist in a gene therapy product. We have tested this notion byexamining the effects of the degree of supercoiling on cationiclipid-mediated gene transfer in vitro and in vivo. Methods. An ion-exchange high performance liquidchromatography (HPLC) method was developed to accurately quantitatethe relative amounts of supercoiled DNA in purified plasmid. A sample of thepurified plasmid was fully relaxed using topoisomerase. Next, the ability ofvarious levels of supercoiled plasmid to transfect mammalian cells wasmeasured. Results. This study suggests that there is no relationbetween the degree of supercoiling and lipofection efficiency. Subsequenttransfection using several different lipofection agents, different celltypes, and an in vivo model support these results. Conclusions. In considering a specification for the amountof supercoiled plasmid in a gene therapy product, it must be noted that therelaxed forms of the plasmid are no less efficient at gene delivery than thesupercoiled forms.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007531405796
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  • 14
    Electronic Resource
    Electronic Resource
    The reproductive biology ofJaborosa integrifolia (Solanaceae): Why its fruits are so rare? (2000)
    Springer
    Plant systematics and evolution 225 (2000), S. 15-28 
    Details
    ISSN: 1615-6110
    Keywords: Solanaceae ; Jaborosa integrifolia ; stigma-height polymorphism ; pollen tube growth ; breeding system ; self-incompatibility ; nectar sugar composition ; nectar removal effects ; Sphingidae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Jaborosa integrifolia exhibits stigma-height polymorphism. There are individuals with flowers where anthers and stigma are at the same height but the rule is variable herkogamy, the most common type (75%) being that with an exerted stigma. Self- and cross-tubes did not differ in their capability to reach the ovary (t = −0.67,P 〈 0.53); they had a high growth rate (6.95 ± 2.28 mm h−1). There is not autogamy but mostly self-incompatibility. Fruits from controlled cross-pollination showed the highest seed set and seed viability. The nectar sugar is characterized by a similar amount of glucose and fructose, and by the absence of sucrose. Although nectar secretion was continuous throughout the life of the flower, most nectar was secreted during the first 24 h after flower opening. Nectar production costs appear to be lower than in other species since nectar secretion is neither inhibited after a removal (i.e. a pollinator visit) nor reabsorbed as the flower ages. Sphingids visit the flowers mainly after midnight. They insert their proboscis down to the base of the corolla tube to reach the nectar. The upper limit to fruit production is set by pollinator visits. Fruits produced from open-pollinated flowers are often predated by numerous larvae (mainly lepidopteran ones). Considering that this species is mostly self-incompatible and pollination is limited, that each plant displays only a low number of flowers throughout the flowering season, and that there is a high rate of fruit predation, it is not surprising that fruits ofJ. integrifolia are so rare.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00985456
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  • 15
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    Functional characterization of the PS II–LHC II supercomplex isolated by a direct method from spinach thylakoid membranes (2000)
    Springer
    Photosynthesis research 64 (2000), S. 179-187 
    Details
    ISSN: 1573-5079
    Keywords: EPR ; fluorescence ; Photosystem II ; thermoluminescence ; thylakoid membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Recently, a novel procedure to isolate a highly pure and active Photosystem II preparation directly from thylakoid membranes, referred to as PS II–LHC II supercomplex, was reported [Eshaghi et al. (1999) FEBS Lett 446: 23–26]. In addition to the reaction center core proteins, the supercomplex contains all the extrinsic proteins of the oxygen evolving complex and a set of chlorophyll a/b binding proteins. In this paper, the functional properties of this isolated supercomplex are further characterized by using EPR spectroscopy, thermoluminescence, fluorescence relaxation kinetics and flash induced oxygen yield measurements. The PS II–LHC II supercomplex contains, in addition to QA and QB, a small pool of plastoquinone (PQ). Although the isolated complex is no longer membrane bound, it has preserved functional characteristics of a well defined PS II preparation with the exception of some modification of QB sites.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006404302573
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  • 16
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    Solubilization of green plant thylakoid membranes with n-dodecyl-α,D-maltoside. Implications for the structural organization of the Photosystem II, Photosystem I, ATP synthase and cytochrome b6 f complexes (2000)
    Springer
    Photosynthesis research 64 (2000), S. 155-166 
    Details
    ISSN: 1573-5079
    Keywords: CF0F1 ; cytochrome b 6 f ; electron microscopy ; grana ; Photosystem I ; Photosystem II
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A biochemical and structural analysis is presented of fractions that were obtained by a quick and mild solubilization of thylakoid membranes from spinach with the non-ionic detergent n-dodecyl-α,D-maltoside, followed by a partial purification using gel filtration chromatography. The largest fractions consisted of paired, appressed membrane fragments with an average diameter of about 360 nm and contain Photosystem II (PS II) and its associated light-harvesting antenna (LHC II), but virtually no Photosystem I, ATP synthase and cytochrome b 6 f complex. Some of the membranes show a semi-regular ordering of PS II in rows at an average distance of about 26.3 nm, and from a partially disrupted grana membrane fragment we show that the supercomplexes of PS II and LHC II represent the basic structural unit of PS II in the grana membranes. The numbers of free LHC II and PS II core complexes were very high and very low, respectively. The other macromolecular complexes of the thylakoid membrane occurred almost exclusively in dispersed forms. Photosystem I was observed in monomeric or multimeric PS I-200 complexes and there are no indications for free LHC I complexes. An extensive analysis by electron microscopy and image analysis of the CF0F1 ATP synthase complex suggests locations of the δ (on top of the F1 headpiece) and ∈ subunits (in the central stalk) and reveals that in a substantial part of the complexes the F1 headpiece is bended considerably from the central stalk. This kinking is very likely not an artefact of the isolation procedure and may represent the complex in its inactive, oxidized form.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006476213540
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  • 17
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    Selective replacement of the active and inactive pheophytin in reaction centres of Photosystem II by 131-deoxo-131-hydroxy-pheophytin a and comparison of their 6 K absorption spectra (2000)
    Springer
    Photosynthesis research 64 (2000), S. 189-198 
    Details
    ISSN: 1573-5079
    Keywords: Photosystem II ; pheophytin a ; pigment exchange ; reaction centre ; 131-deoxo-131-hydroxy pheophytin a
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pheophytin a (Pheo) in Photosystem II reaction centres was exchanged for 131-deoxo-131-hydroxy-pheophytin a (131-OH-Pheo). The absorption bands of 131-OH-Pheo are blue-shifted and well separated from those of Pheo. Two kinds of modified reaction centre preparations can be obtained by applying the exchange procedure once (RC1×) or twice (RC2×). HPLC analysis and Pheo QX absorption at 543 nm show that in RC1× about 50% of Pheo is replaced and in RC2× about 75%. Otherwise, the pigment and protein composition are not modified. Fluorescence emission and excitation spectra show quantitative excitation transfer from the new pigment to the emitting chlorophylls. Photoaccumulation of Pheo− is unmodified in RC1× and decreased only in RC2×, suggesting that the first exchange replaces the inactive and the second the active Pheo. Comparing the effects of the first and the second replacement on the absorption spectrum at 6 K did not reveal substantial spectral differences between the active and inactive Pheo. In both cases, the absorption changes in the QY region can be interpreted as a combination of a blue shift of a transition at 684 nm, a partial decoupling of chlorophylls absorbing at 680 nm and a disappearance of Pheo absorption in the 676-680 nm region. No absorption decrease is observed at 670 nm for RC1× or RC2×, showing that neither of the two reaction centre pheophytins contributes substantially to the absorption at this wavelength.
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    URL: http://dx.doi.org/10.1023/A:1006407314449
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    Excitation energy transfer in aggregates of Photosystem I and Photosystem II of the cyanobacterium Synechocystis sp. PCC 6803: Can assembly of the pigment-protein complexes control the extent of spillover? (2000)
    Springer
    Photosynthesis research 64 (2000), S. 199-207 
    Details
    ISSN: 1573-5079
    Keywords: cyanobacteria ; excitation energy transfer ; membrane proteins assembly ; Photosystem I ; Photosystem II ; spillover ; state transitions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The fluorescence profile of Photosystem I/Photosystem II mixtures in different solvent systems shows that both non-hydrophobic and hydrophobic interactions govern their association and control energy transfer from Photosystem II to Photosystem I. The non-hydrophobic interactions lead to a highly efficient excitation energy transfer from Photosystem II to Photosystem I. In view of this, we propose that similar non-hydrophobic interactions, between the Photosystem II and Photosystem I peripheral proteins, also play a significant role in their association in thylakoids that control state transitions in cyanobacteria.
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    URL: http://dx.doi.org/10.1023/A:1006485823403
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    Light-induced quenching of chlorophyll fluorescence at 77 K in leaves, chloroplasts and Photosystem II particles (2000)
    Springer
    Photosynthesis research 65 (2000), S. 219-229 
    Details
    ISSN: 1573-5079
    Keywords: energy dissipation ; Photosystem I ; Photosystem II ; spectroscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The light-induced chlorophyll (Chl) fluorescence decline at 77 K was investigated in segments of leaves, isolated thylakoids or Photosystem (PS) II particles. The intensity of chlorophyll fluorescence declines by about 40% upon 16 min of irradiation with 1000 μmol m−2 s−1 of white light. The decline follows biphasic kinetics, which can be fitted by two exponentials with amplitudes of approximately 20 and 22% and decay times of 0.42 and 4.6 min, respectively. The decline is stable at 77 K, however, it is reversed by warming of samples up to 270 K. This proves that the decline is caused by quenching of fluorescence and not by pigment photodegradation. The quantum yield for the induction of the fluorescence decline is by four to five orders lower than the quantum yield of QA reduction. Fluorescence quenching is only slightly affected by addition of ferricyanide or dithionite which are known to prevent or stimulate the light-induced accumulation of reduced pheophytin (Pheo). The normalised spectrum of the fluorescence quenching has two maxima at 685 and 695 nm for PS II emission and a plateau for PS I emission showing that the major quenching occurs within PS II. ‘Light-minus-dark’ difference absorbance spectra in the blue spectral region show an electrochromic shift for all samples. No absorbance change indicating Chl oxidation or Pheo reduction is observed in the blue (410–600 nm) and near infrared (730–900 nm) spectral regions. Absorbance change in the red spectral region shows a broad-band decrease at approximately 680 nm for thylakoids or two narrow bands at 677 and 670–672 nm for PS II particles, likely resulting also from electrochromism. These absorbance changes follow the slow component of the fluorescence decline. No absorbance changes corresponding to the fast component are found between 410 and 900 nm. This proves that the two components of the fluorescence decline reflect the formation of two different quenchers. The slow component of the light-induced fluorescence decline at 77 K is related to charge accumulation on a non-pigment molecule of the PS II complex.
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    URL: http://dx.doi.org/10.1023/A:1010730103382
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    Low-frequency fourier transform infrared spectroscopy of the oxygen-evolving complex in Photosystem II* (2000)
    Springer
    Photosynthesis research 66 (2000), S. 57-63 
    Details
    ISSN: 1573-5079
    Keywords: FTIR ; manganese cluster ; oxygen-evolving complex ; Photosystem II ; plastoquinone ; vibrational spectroscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this communication, we report our progress on the development of low-frequency Fourier transform infrared (FTIR) spectroscopic techniques to study metal-substrate and metal-ligand vibrational modes in the Photosystem II/oxygen-evolving complex (PS II/OEC). This information will provide important structural and mechanistic insight into the OEC. Strong water absorption in the low-frequency region (below 1000 cm−1), a lack of suitable materials, and temperature control problems have limited previous FTIR spectroscopic studies of the OEC to higher frequencies (〉1000 cm−1). We have overcome these technical difficulties that have blocked access to the low-frequency region and have developed successive instruments that allow us to move deeper into the low-frequency region (down to 350 cm−1), while increasing both data accumulation efficiency and S/N ratio. We have detected several low-frequency modes in the S2/S1spectrum that are specifically associated with these two states. Our results demonstrate the utility of FTIR techniques in accessing low-frequency modes in Photosystem II and in proteins generally.
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    URL: http://dx.doi.org/10.1023/A:1010779327960
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    Airborne test of laser pump-and-probe technique for assessment of phytoplankton photochemical characteristics (2000)
    Springer
    Photosynthesis research 66 (2000), S. 45-56 
    Details
    ISSN: 1573-5079
    Keywords: chlorophyll ; energy quenching ; fluorescence ; LIDAR ; photochemistry ; photosynthesis ; Photosystem II ; phytoplankton ; pump and probe ; remote sensing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Initial results of the airborne LIDAR measurement of photochemical quantum yield, ΦPo, and functional absorption cross-section, σPS II, of Photosystem II (PS II) are reported. NASA's AOL3 LIDAR was modified to implement short-pulse pump-and-probe (SP-P&P) LIDAR measurement protocol. The prototype system is capable of measuring a pump-induced increase in probe-stimulated chlorophyll fluorescence, ΔF/Fsat, along with the acquisition of `conventional' LIDAR-fluorosensor products from an operational altitude of 150 m. The use of a PS II sub-saturating probe pulse increases the response signal but also results in excessive energy quenching (EEQ) affecting the ΔF/Fsat magnitude. The airborne data indicated up to a 3-fold EEQ-caused decline in ΔF/Fsat, and 2-fold variability in the EEQ rate constant over a spatial scale a few hundred kilometers. Therefore, continuous monitoring of EEQ parameters must be incorporated in the operational SP-P&P protocol to provide data correction for the EEQ effect. Simultaneous airborne LIDAR measurements of ΦPo and σPS II with EEQ correction were shown to be feasible and optimal laser excitation parameters were determined. Strong daytime ΔF/Fsat decline under ambient light was found in the near-surface water layer over large aquatic areas. An example of SP-P&P LIDAR measurement of phytoplankton photochemical and fluorescent characteristics in the Chesapeake Bay mouth is presented. Prospects for future SP-P&P development and related problems are discussed.
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    URL: http://dx.doi.org/10.1023/A:1010764420934
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    An FTIR study on the structure of the oxygen-evolving Mn-cluster of Photosystem II in different spin forms of the S2 state (2000)
    Springer
    Photosynthesis research 63 (2000), S. 47-57 
    Details
    ISSN: 1573-5079
    Keywords: Fourier transform infrared spectroscopy ; Mn-cluster ; oxygen evolution ; photosynthesis ; Photosystem II ; vibrational spectroscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The S2 state of the oxygen-evolving Mn-cluster of Photosystem II (PS II) is known to have different forms that exhibit the g =2 multiline and g = 4.1 EPR signals. These two spin forms are interconvertible at 〉 200 K and the relative amplitudes of the two signals are dependent on the species of cryoprotectant and alcohol contained in the medium. Also, it was recently found that the mutiline form can be converted to the g = 4.1 form by absorption of near-infrared light by the Mn-cluster itself at around 150 K [Boussac et al. (1996) Biochemistry 35: 6984–6989]. We have used light-induced Fourier transform infrared (FTIR) difference spectroscopy to study the structural difference in these two S2 forms. FTIR difference spectra for S2/S1 as well as for S2QA -/S1QA measured at cryogenic temperatures using PS II membranes in the presence of various cryoprotectants, and monohydric alcohols did not show any specific differences except for intensities of amide I bands, which were larger when ethylene glycol or glycerol was present in addition to sucrose. This result was interpreted due to more flexible movement of the protein backbones upon S2 formation with a higher cryoprotectant content. Light-induced difference spectra measured at 150 K using either blue light without near-infrared light or red plus near-infrared light also did not show any detectable difference. In addition, a different spectrum upon near-infrared illumination at 150 K of the PS II sample in which the S2 state had been photogenerated at 200 K exhibited no meaningful signals. These results indicate that the two S2 forms that give rise to the multiline and g = 4.1 signals have only minor differences, if any, in the structures of amino-acid ligands and polypeptide backbones. This conclusion suggests that conversion between the two spin states is caused by a spin-state transition in the Mn(III) ion rather than valence swapping within the Mn-cluster that would considerably affect the vibrations of ligands.
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    URL: http://dx.doi.org/10.1023/A:1006362118267
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    Short-pulse pump-and-probe technique for airborne laser assessment of Photosystem II photochemical characteristics (2000)
    Springer
    Photosynthesis research 66 (2000), S. 33-44 
    Details
    ISSN: 1573-5079
    Keywords: chlorophyll ; fluorescence ; LIDAR ; photosynthesis ; Photosystem II ; pump and probe ; remote sensing ; singlet-singlet quenching ; singlet-triplet quenching
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The development of a technique for laser measurement of fPhotosystem II (PS II) photochemical characteristics of phytoplankton and terrestrial vegetation from an airborne platform is described. Results of theoretical analysis and experimental study of pump-and-probe measurement of the PS II functional absorption cross-section and photochemical quantum yield are presented. The use of 10 ns probe pulses of PS II sub-saturating intensity provides a significant, up to 150-fold, increase in the fluorescence signal compared to conventional `weak-probe' protocol. Little effect on the fluorescence yield from the probe-induced closure of PS II reaction centers is expected over the short pulse duration, and thus a relatively intense probe pulse can be used. On the other hand, a correction must be made for the probe-induced carotenoid triplet quenching and singlet-singlet annihilation. A Stern-Volmer model developed for this correction assumes a linear dependence of the quenching rate on the laser pulse fluence, which was experimentally validated. The PS II saturating pump pulse fluence (532 nm excitation) was found to be 10 and 40 μmol quanta m−2 for phytoplankton samples and leaves of higher plants, respectively. Thirty μs was determined as the optimal delay in the pump-probe pair. Our results indicate that the short-pulse pump-and-probe measurement of PS II photochemical characteristics can be implemented from an airborne platform using existing laser and LIDAR technologies.
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    URL: http://dx.doi.org/10.1023/A:1010795820025
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    Characterization of carotenes in a combination of a C18 HPLC column with isocratic elution and absorption spectra with a photodiode-array detector (2000)
    Springer
    Photosynthesis research 65 (2000), S. 93-99 
    Details
    ISSN: 1573-5079
    Keywords: absorption spectrum ; carotene ; carotenoid ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Carotenes have attracted much attention in recent years for their biological function in processes such as photosynthesis. The characterization of carotenes is difficult, however, because they consist of only carbon and hydrogen atoms, without oxygen. In the present study, we systematically examined the chemical structures of more than 30 carotenes, including most of the carotenes found in phototrophic organisms, and observed their elution order using a Novapak C18 HPLC column with simple isocratic elution. The elution order of the carotenes was C30, C40,C45 then C50. The C40 carotenes with fewer conjugated double bonds (N) had longer retention times. With respect to the end groups, the carotenes eluted in the following order: φ, Ψ, ∈ then β end groups. Furthermore, absorption spectra in the HPLC eluent used were recorded with a photodiode-array detector. A greater N value was associated with a longer absorption maximum wavelength. Since the conjugated end groups (φ and β) influenced the absorption spectra and the non-conjugated end groups (Ψ and ∈) did not, the number of conjugated end groups (zero, one and two) was clearly distinguishable. Therefore, the chemical structures of carotenes can be easily determined by a combination of the HPLC retention times and the absorption spectra.
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    URL: http://dx.doi.org/10.1023/A:1006445503030
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    Low resistance against novel 2-benzylamino-1,3,5-triazine herbicides in atrazine-resistant Chenopodium album plants (2000)
    Springer
    Photosynthesis research 65 (2000), S. 115-120 
    Details
    ISSN: 1573-5079
    Keywords: electron transport ; herbicides ; novel triazines ; O-J-I-P fluorescence transient ; Photosystem II ; resistance ; thylakoid membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of nine novel 2-benzylamino-1,3,5-triazines on photosynthetic reactions were measured in thylakoids isolated from wild-type and atrazine-resistant plants of Chenopodium album. The resistant plants have a mutation of serine for glycine at position 264 of the D1 protein. The measurement of oxygen evolution and chlorophyll a fluorescence induction indicated a 2–4-fold stronger inhibition by the 6-trifluoromethyl analogues of Photosystem II-dependent electron flow than atrazine. Analogues having a 6-methyl-, 6-monofluoromethyl or 6-difluoromethyl substitution were weak inhibitors, indicating that the 6-trifluoro group is very important for strong inhibition. All the nine novel 2-benzylamino-1,3,5-triazines were almost as active in wild-type as in atrazine-resistant thylakoids, indicating that the benzylamino substitution may be important for the lack of resistance in the atrazine-resistant plants.
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    URL: http://dx.doi.org/10.1023/A:1006493703939
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    Nitrogen ligation to the manganese cluster of Photosystem II in the absence of the extrinsic proteins and as a function of pH (2000)
    Springer
    Photosynthesis research 65 (2000), S. 175-185 
    Details
    ISSN: 1573-5079
    Keywords: electron paramagnetic resonance ; extrinsic proteins ; manganese cluster ; oxygen evolution ; photosynthesis ; Photosystem II
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three extrinsic proteins (PsbO, PsbP and PsbQ), with apparent molecular weights of 33, 23 and 17 kDa, bind to the lumenal side of Photosystem II (PS II) and stabilize the manganese, calcium and chloride cofactors of the oxygen evolving complex (OEC). The effect of these proteins on the structure of the tetramanganese cluster, especially their possible involvement in manganese ligation, is investigated in this study by measuring the reported histidine-manganese coupling [Tang et al. (1994) Proc Natl Acad Sci USA 91: 704–708] of PS II membranes depleted of none, two or three of these proteins using ESEEM (electron spin echo envelope modulation) spectroscopy. The results show that neither of the three proteins influence the histidine ligation of manganese. From this, the conserved histidine of the 23 kDa protein can be ruled out as a manganese ligand. Whereas the 33 and 17 kDa proteins lack conserved histidines, the existence of a 33 kDa protein-derived carboxylate ligand has been posited; our results show no evidence for a change of the manganese co-ordination upon removal of this protein. Studies of the pH-dependence of the histidine–manganese coupling show that the histidine ligation is present in PS II centers showing the S2 multiline EPR signal in the pH-range 4.2–9.5.
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    URL: http://dx.doi.org/10.1023/A:1006435432185
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    Degradation of the Photosystem II D1 and D2 proteins in different strains of the cyanobacterium Synechocystis PCC 6803 varying with respect to the type and level of psbA transcript (2000)
    Springer
    Plant molecular biology 42 (2000), S. 635-645 
    Details
    ISSN: 1573-5028
    Keywords: cyanobacteria ; D1 ; D2 ; Photosystem II ; psbA ; Synechocystis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The turnover of the D1 and D2 proteins of Photosystem II (PSII) has been investigated by pulse-chase radiolabeling in several strains of the cyanobacterium Synechocystis PCC 6803 containing different types and levels of the psbA transcript. Strains lacking psbA1 and psbA3 gene and containing high levels of the psbA2 transcript showed the selective synthesis of D1 whose degradation could be slowed down by the protein synthesis inhibitor lincomycin. In contrast, in strains containing just the psbA3 gene, the intensity of the D1 protein labeling was lower and labeling of the D2 and CP43 proteins was stimulated in comparison to the psbA2-containing strains. In addition, the rate and selectivity of the D1 degradation and its dependence on the presence of lincomycin was proportional to the level of the psbA3 transcript in the particular strain. Consequently, there was parallel, lincomycin-independent and slowed-down breakdown of the D1 and D2 proteins in strains with the lowest level of psbA3 transcript. These results are discussed in terms of a model in which the rate of D1 and D2 degradation in cyanobacteria is affected not only by the rate of PSII photodamage, but also by the availability of newly synthesized D1 protein. Moreover, the comparison of the non-oxygen-evolving D1 mutants D170A** and Y161F*** differing by the presence of tyrosine Z has indicated a minor role of the oxidized form of this secondary PSII electron donor in the donor side mechanism of D1 and D2 protein breakdown.
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    URL: http://dx.doi.org/10.1023/A:1006305308196
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    Selective Phytotoxic Activity of Withanolides from Iochroma australe to Crop and Weed Species (2000)
    Springer
    Journal of chemical ecology 26 (2000), S. 2187-2196 
    Details
    ISSN: 1573-1561
    Keywords: Phytotoxicity ; allelopathy ; withanolides ; Iochroma australe ; Solanaceae ; weed ; Sorghum halepense ; Chenopodium album ; crop ; Lactuca sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Phytotoxic effects of Iochroma australe extract and 4,7,20-oxowithanolides were examined in Petri dish bioassays at 10, 100, and 1000 ppm. The extract and the major constituent (17S,20R,22R)-4β,7β,20α-trihydroxy-1oxowitha-2,5,24-trienolide (2) reduced growth of the radicle of the weeds Sorghum halepense (Monoct.) and Chenopodium album (Dicot.). Neither the extract nor withanolides had significant effect on germination or radicle length of the commercial crop species Lactuca sativa. From our experimental data we conclude that the withanolides tested here are natural products with selective herbicidal activity against weed species.
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    URL: http://dx.doi.org/10.1023/A:1005576617857
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    Identification of 1-Octacosanal and 6-Methoxy-2-Benzoxazolinone from Wheat as Ovipositional Stimulants for Hessian Fly, Mayetiola destructor (2000)
    Springer
    Journal of chemical ecology 26 (2000), S. 859-873 
    Details
    ISSN: 1573-1561
    Keywords: Cecidomyiidae ; insect behavior ; egg-laying ; chemoreception ; MBOA ; Gramineae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Bioassay-guided fractionations of a dichloromethane extract of wheat epicuticular wax allowed the identification of 1-octacosanal and 6-methoxy-2-benzoxazolinone (MBOA) as the major components that stimulate oviposition by the Hessian fly, Mayetiola destructor. These compounds were identified by their mass spectral fragmentation patterns and by comparison of their gas chromatographic retention times with synthetic samples. Synthetic samples of 1-octacosanal or MBOA stimulated significant oviposition when compared with solvent controls. In combination, these compounds elicited a synergistic effect on the number of eggs laid by females compared to when they were presented alone. In a choice bioassay, a mixture of synthetic 1-octacosanal and MBOA in the approximate concentrations determined to be present in one plant equivalent of crude extract stimulated the same amount of oviposition as one plant equivalent of extract. This showed that together these two compounds appear to be responsible for the major proportion of the ovipositional stimulatory activity of the wheat epicuticular wax extract. Comparison of the activity of five straight-chain primary aldehydes with chain lengths from C22 to C30 revealed a relationship between chain length and the number of eggs laid by female Hessian flies, with 1-hexacosanal and 1-heptacosanal the most active of the aldehydes tested.
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    URL: http://dx.doi.org/10.1023/A:1005499907009
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    Effect of Solanum Glycoalkaloids on Potato Aphid, Macrosiphum euphorbiae: Part II (2000)
    Springer
    Journal of chemical ecology 26 (2000), S. 1113-1121 
    Details
    ISSN: 1573-1561
    Keywords: Macrosiphum euphorbiae ; potato aphid ; Homoptera ; Aphididae ; Solanaceae ; glycoalkaloids ; bioassay ; commersonine ; demissine ; solamargine ; solasodine ; solasonine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We report the deterrent, toxic, and antireproductive effect of several Solanum glycoalkaloids on the potato aphid, Macrosiphum euphorbiae, and discuss the structure–activity relationship of the tested compounds. The results indicate a structure-dependent biological effect of the glycoalkaloids, suggesting that, while the structure of the aglycone defines a basal biological effect, the carbohydrate moiety is crucial for the overall biological effect.
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    URL: http://dx.doi.org/10.1023/A:1005471624833
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    Determination of the Tropospheric Hydroxyl Radical by Liquidphase Scrubbing and HPLC: Preliminary Results (2000)
    Springer
    Journal of atmospheric chemistry 36 (2000), S. 81-105 
    Details
    ISSN: 1573-0662
    Keywords: hydroxyl radical ; OH ; HPLC ; chromatography ; atmosphere ; air ; troposphere ; determination ; analysis ; air scrubbing ; scavenging ; fluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract A preliminary study was carried out toexamine the feasibility of measuring tropospherichydroxyl radicals (OH) by liquidphase scrubbing andhigh performance liquid chromatography (HPLC). Thepotential advantages of this approach are itssimplicity, portability, and low expense. Thesampling system employs glass bubblers to trapatmospheric OH into a buffered solution of salicylicacid (o-hydroxybenzoic acid, OHBA). Rapidreaction of OH with OHBA produces a stable fluorescentproduct, 2,5-dihydroxybenzoic acid (2,5-DHBA), whichis determined by reverse-phase HPLC and fluorescencedetection. Our preliminary field results indicatethat this method is most suitable for OH measurementsin clean tropospheric air, where interferences fromother atmospheric species appear to be negligible orminor relative to polluted air. In clean air, thesampling period is about 45–90 minutes, which yieldsa detection limit of approximately 3–6 ×105 radicalscm-3. During an OHintercomparison experiment at the Caribou samplingsite in Colorado, our liquidphase scrubber method wascompared with the ion-assisted mass spectrometry (MS)method. Our results were within the same range asthose of the ion-assisted MS method (1–5 ×106 radicals cm-3) within our precision atthat time (about ±30–50%). Preliminary testsin Pullman, WA indicated that the method might alsofunction in moderately polluted air by acidifying thescrubbing solution or by adding a scavenger tosuppress interferences. In Pullman, mid-day OHconcentrations were usually in the range of 2–20 ×106 radicals cm-3. Nighttime OHconcentrations were always low, either at or slightlyabove the detection limit.
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    URL: http://dx.doi.org/10.1023/A:1006313021834
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    Electron spin-lattice relaxation studies of different forms of the S2 state multiline EPR signal of the Photosystem II oxygen-evolving complex (2000)
    Springer
    Photosynthesis research 66 (2000), S. 189-198 
    Details
    ISSN: 1573-5079
    Keywords: electron spin-lattice relaxation rate ; manganese ; oxygen-evolving complex ; photosynthesis ; Photosystem II
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The pulsed EPR inversion recovery sequence has been utilized to monitor the temperature dependence of the electron spin-lattice relaxation rate of the Mn cluster of the Photosystem II oxygen evolving complex poised in a variety of S 2 state forms giving rise to g = 2 multiline EPR signals. A previous study (Lorigan and Britt (1994) Biochemistry 33: 12072–12076) showed that for PS II membranes treated with 5% ethanol, the S 2 state Mn cluster relaxes via the Orbach spin-lattice relaxation mechanism, where the relaxation is enhanced via phonon scattering off an excited state spin manifold, in this case at an energy of Δ = 36.5 cm−1 above the S = 1/2 ground state giving rise to the multiline EPR signal. Parallel experiments are reported for PS II membranes with 5% methanol, treated with ammonia, and following short and long term dark adaptation. In each case, the temperature dependence of the electron spin-lattice relaxation rate is consistent with Orbach relaxation, and the range of excited state energies is relatively narrow (33.8 cm−1 ≤ Δ ≤ 39.7 cm−1). In addition, short term dark adapted (6 min, ‘active state’) PS II membranes show biphasic recovery traces which indicate that a minority fraction of the oxygen evolving complexes are trapped in a form with greatly slowed spin-lattice relaxation.
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    URL: http://dx.doi.org/10.1023/A:1010604607891
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    Embryo rescue of interspecific hybrids of Brachiaria spp (2000)
    Springer
    Plant cell, tissue and organ culture 61 (2000), S. 175-182 
    Details
    ISSN: 1573-5044
    Keywords: adventitious shoots ; Gramineae ; immature embryos ; micropropagation ; somatic embryogenesis ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Age of explant and six different media were evaluated with the objective of regenerating higher numbers of interspecific hybrids between sexual and apomictic Brachiaria. Immature embryos of 7–8, 9–10 and 11–12 days after pollination (DAP), from artificial hybridization between Brachiaria ruziziensis (R) as female parent, and B. brizantha (B) or B. decumbens (D) as male parent, were cultured in modified MS media (M4) – supplemented with different combinations of growth regulators and vitamins. Embryos cultured 9–12 DAP showed high percentage (85–100%) of germination for all the crosses examined. Germination and survival rates varied according to accessions within crosses. Six different media (all modified MS with different growth regulators and vitamins) were tested with the objective of inducing multiple shoots from 7 to 10 DAP embryos, from crosses between R × B. The media M1, supplemented with Kinetin (13.94 μM) and NAA (5.37 μM), and media M3, supplemented with BA (4.44 μM and IAA 2.85 μM), regenerated adventitious shoots and calli about 30–40 days after inoculation. The highest multiplication rate observed was 2.85 shoots per explant in media M1, 60–70 days after culturing. Two other media, M6, supplemented with 2,4-D (13.57 μM) and M2, with 2,4-D (9.05 μM) and BA (8.87 μM) exclusively induced the formation of calli. The described protocols proved to be efficient in regenerating healthy seedlings from immature embryos of interspecific hybrids in Brachiaria.
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    Effects of cytokinin/auxin combinations on organogenesis, shoot regeneration and tropane alkaloid production in Duboisia myoporoides (2000)
    Springer
    Plant cell, tissue and organ culture 62 (2000), S. 125-133 
    Details
    ISSN: 1573-5044
    Keywords: Duboisia myoporoides ; plant growth regulators ; shoot culture ; Solanaceae ; tropane alkaloid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Leaf explants of Duboisia myoporoides R. Br. were exposed to 65 different cytokinin/auxin combinations and the morphogenetic responses compared. Two different types of calluses were induced depending on the cytokinin/auxin combinations used for callus induction. Non-rooted shoots were regenerated from calluses induced with 7 different cytokinin/auxin combinations, cultured in the regeneration medium and also from the calluses induced with 2 different cytokinin/auxin combinations, cultured in the callus induction medium. Shoot-bud regenerating ability of the calluses, number of leaves formed/shoot, lengths of the leaves and shoots and tropane alkaloid biosynthetic ability of the non-rooted shoots depended on the cytokinin/auxin combinations used at the callus induction stage. Hyoscyamine and scopolamine were detected in the 11-week-old callus induced with BA 10 μM + NAA 1 μM. These alkaloids were also detected in the 6 and 9-week-old non-rooted shoots regenerated from callus induced with BA 10 μM + NAA 1 μM, cultured in regeneration medium and from the callus induced with BA 10 μM + IBA 0.1 μM, cultured in callus induction medium. Shoots were regenerated in liquid medium supplemented with BA 10 μM + IBA 0.1 μM. No tropane alkaloids were detected in the 4-week-old shoot cultured in liquid medium.
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    Reduction of QA in the dark: Another cause of fluorescence Fo increases by high temperatures in higher plants (2000)
    Springer
    Photosynthesis research 63 (2000), S. 23-34 
    Details
    ISSN: 1573-5079
    Keywords: chlororespiration ; pheophytin a ; photosynthesis ; Photosystem II ; potato ; tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Increases in the chlorophyll fluorescence Fo (dark level fluorescence) during heat treatments were studied in various higher plants. Besides the dissociation of light-harvesting chlorophyll a/b protein complexes from the reaction center complex of PS II and inactivation of PS II, dark reduction of QA via plastoquinone (PQ) seemed to be related to the Fo increase at high temperatures. In potato leaves or green tobacco cultured cells, a part of the Fo increase was quenched by light, reflecting light-induced oxidation of QA - which had been reduced in the dark at high temperatures. Appearance of the Fo increase due to QA reduction depended on the plant species, and the mechanisms for this are proposed. The reductants seemed to be already present and formed by very brief illumination of the leaves at high temperatures. A ndhB-less mutant of tobacco showed that complex I type NAD(P)H dehydrogenase is not involved in the heat-induced reduction of QA. Quite strong inhibition of the QA reduction by diphenyleneiodonium suggests that a flavoenzyme is one of the electron mediator to PQ from the reductant in the stroma. Reversibility of the heat-induced QA reduction suggests that an enzyme(s) involved is activated at high temperatures and mostly returns to an inactive form at room temperature (25 °C).
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    The IdiA protein of Synechococcus sp. PCC 7942 functions in protecting the acceptor side of Photosystem II under oxidative stress (2000)
    Springer
    Photosynthesis research 63 (2000), S. 145-157 
    Details
    ISSN: 1573-5079
    Keywords: cyanobacteria ; IdiA ; oxidative stress ; Photosystem II ; PsbO ; Synechococcus sp. PCC 7942 and PCC 6301
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Synechococcus sp. strains PCC 7942 and PCC 6301 contain a 35 kDa protein called IdiA (Iron deficiency induced protein A) that is expressed in elevated amounts under Fe deficiency and to a smaller extent also under Mn deficiency. Absence of this protein was shown to mainly damage Photosystem II. To decide whether IdiA has a function in optimizing and/or protecting preferentially either the donor or acceptor side reaction of Photosystem II, a comparative analysis was performed of Synechococcus sp. PCC 7942 wild-type, the IdiA-free mutant, the previously constructed PsbO-free Synechococcus PCC 7942 mutant and a newly constructed Synechococcus PCC 7942 double mutant lacking both PsbO and IdiA. Measurements of the chlorophyll fluorescence and determinations of Photosystem II activity using a variety of electron acceptors gave evidence that IdiA has its main function in protecting the acceptor side of Photosystem II. Especially, the use of dichlorobenzoquinone, preferentially accepting electrons from QA, gave a decreased O2 evolving activity in the IdiA-free mutant. Investigations of the influence of hydrogen peroxide treatment on cells revealed that this treatment caused a significantly higher damage of Photosystem II in the IdiA-free mutant than in wild-type. These results suggest that although the IdiA protein is not absolutely required for Photosystem II activity in Synechococcus PCC 7942, it does play an important role in protecting the acceptor side against oxidative damage.
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    Investigation of the plastoquinone pool size and fluorescence quenching in thylakoid membranes and Photosystem II (PS II) membrane fragments (2000)
    Springer
    Photosynthesis research 63 (2000), S. 171-182 
    Details
    ISSN: 1573-5079
    Keywords: Chlorophyll fluorescence ; non-photochemical quenching ; Photosystem II ; thylakoid membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The efficiency of oxidized endogenous plastoquinone-9 (PQ-9) as a non-photochemical quencher of chlorophyll fluorescence has been analyzed in spinach thylakoids and PS II membrane fragments isolated by Triton X-100 fractionation of grana stacks. The following results were obtained: (a) After subjection of PS II membrane fragments to ultrasonic treatment in the presence of PQ-9, the area over the induction curve of chlorophyll fluorescence owing to actinic cw light increases linearly with the PQ-9/PS II ratio in the reconstitution assay medium; (b) the difference of the maximum fluorescence levels, Fmax, of the induction curves, measured in the absence and presence of DCMU, is much more pronounced in PS II membrane fragments than in thylakoids; (c) the ratio Fmax(-DCMU)/Fmax(+DCMU) increases linearly with the content of oxidized PQ-9 that is varied in the thylakoids by reoxidation of the pool after preillumination and in PS II membrane fragments by the PQ-9/PS II ratio in the reconstitution assay; (d) the reconstitution procedure leads to tight binding of PQ-9 to PS II membrane fragments, and PQ-9 cannot be replaced by other quinones; (e) the fluorescence quenching by oxidized PQ-9 persists at low temperatures, and (f) oxidized PQ-9 preferentially affects the F695 of the fluorescence emission spectrum at 77 K. Based on the results of this study the oxidized PQ-9 is inferred to act as a non-photochemical quencher via a static mechanism. Possible implications for the nature of the quenching complex are discussed.
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    Inhibition of Photosystem II activity by saturating single turnover flashes in calcium-depleted and active Photosystem II (2000)
    Springer
    Photosynthesis research 63 (2000), S. 209-216 
    Details
    ISSN: 1573-5079
    Keywords: Ca2+-depletion ; charge recombination ; photoinhibition ; Photosystem II ; QA midpoint potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Inhibition of Photosystem II (PS II) activity induced by continuous light or by saturating single turnover flashes was investigated in Ca2+-depleted, Mn-depleted and active PS II enriched membrane fragments. While Ca2+- and Mn-depleted PS II were more damaged under continuous illumination, active PS II was more susceptible to flash-induced photoinhibition. The extent of photoinactivation as a function of the duration of the dark interval between the saturating single turnover flashes was investigated. The active centres showed the most photodamage when the time interval between the flashes was long enough (32 s) to allow for charge recombination between the S2 or S3 and QB − to occur. Illumination with groups of consecutive flashes (spacing between the flashes 0.1 s followed by 32 s dark interval) resulted in a binary oscillation of the loss of PS II-activity in active samples as has been shown previously (Keren N, Gong H, Ohad I (1995), J Biol Chem 270: 806–814). Ca2+- and Mn-depleted PS II did not show this effect. The data are explained by assuming that charge recombination in active PS II results in a back reaction that generates P680 triplet and thence singlet oxygen, while in Ca2+- and Mn-depleted PS II charge recombination occurs through a different pathway, that does not involve triplet generation. This correlates with an up-shift of the midpoint potential of QA in samples lacking Ca2+ or Mn that, in term, is predicted to result in the triplet generating pathway becoming thermodynamically less favourable (G.N. Johnson, A.W. Rutherford, A. Krieger, 1995, Biochim. Biophys. Acta 1229, 201–207). The diminished susceptibility to flash-induced photoinhibition in Ca2+- and Mn-depleted PS II is attributed at least in part to this mechanism.
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    Primary charge separation in Photosystem II (2000)
    Springer
    Photosynthesis research 63 (2000), S. 195-208 
    Details
    ISSN: 1573-5079
    Keywords: charge separation ; disorder ; exciton interaction ; Photosystem II ; reaction center
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In this Minireview, we discuss a number of issues on the primary photosynthetic reactions of the green plant Photosystem II. We discuss the origin of the 683 and 679 nm absorption bands of the PS II RC complex and suggest that these forms may reflect the single-site spectrum with dominant contributions from the zero-phonon line and a pronounced ∼80 cm−1 phonon side band, respectively. The couplings between the six central RC chlorins are probably very similar and, therefore, a `multimer' model arises in which there is no `special pair' and in which for each realization of the disorder the excitation may be dynamically localized on basically any combination of neighbouring chlorins. The key features of our model for the primary reactions in PS II include ultrafast (〈500 fs) energy transfer processes within the multimer, `slow' (∼20 ps) energy transfer processes from peripheral RC chlorophylls to the RC multimer, ultrafast charge separation (〈500 fs) with a low yield starting from the singlet-excited `accessory' chlorophyll of the active branch, cation transfer from this `accessory' chlorophyll to a `special pair' chlorophyll and/or charge separation starting from this `special pair' chlorophyll (∼8 ps), and slow relaxation (∼50 ps) of the radical pair by conformational changes of the protein. The charge separation in the PS II RC can probably not be described as a simple trap-limited or diffusion-limited process, while for the PS II core and larger complexes the transfer of the excitation energy to the PS II RC may be rate limiting.
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    Functional analysis of the PsbX protein by deletion of the corresponding gene in Synechocystis sp. PCC 6803 (2000)
    Springer
    Plant molecular biology 44 (2000), S. 815-827 
    Details
    ISSN: 1573-5028
    Keywords: 4.1 kDa protein ; low-molecular-mass proteins ; oxygen evolution ; Photosystem II ; PSII ; PsbX
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The psbX gene (sml0002) coding for a 4.1 kDa protein in Photosystem II of plants and cyanobacteria was deleted in both wild type and in a Photosystem I-less mutant of the cyanobacterium Synechocystis sp. PCC 6803. Polymerase chain reaction and sequencing analysis showed that the mutants had completely segregated. Deletion of the PsbX protein does not seem to influence growth rate, electron transport or water oxidation ability. Whereas a high light induction of the psbX mRNA could be observed in wild type, deletion of the gene did not lead to high light sensibility. Light saturation measurements and 77K fluorescence measurements indicated a minor disconnection of the antenna in the deletion mutant. Furthermore, fluorescence induction measurements as well as immuno-staining of the D1 protein showed that the amount of Photosystem II complexes in the mutants was reduced by 30%. Therefore, PsbX does not seem to be necessary for the Photosystem II electron transport, but directly or indirectly involved in the regulation of the amount of functionally active Photosystem II centres in Synechocystis sp. PCC 6803.
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    HPLC Screening and GC-MS Confirmation of Triazine Herbicides Residues in Drinking Water from Sugar Cane Area in Brazil (2000)
    Springer
    Water, air & soil pollution 118 (2000), S. 329-338 
    Details
    ISSN: 1573-2932
    Keywords: ametryn ; atrazine ; GC-MS ; HPLC ; simazine ; water
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The extensive use of chlorotriazines as selectiveherbicides in agriculture and their relatively highpersistence imply that these compounds are now presentin the environment, contaminating surface and groundwater. In European countries, United States andCanada, the drinking water ordinance demands a limitedconcentration of 0.5 μg L-1 for the sum of allpesticides and 0.1 μg L-1 with respect to eachcompound, implying on the necessity of sensitive andselective analytical methods. In the present study wedescribe two methods for the analysis of atrazine,simazine and ametryn residues in surface and groundwater collected from the Espraiado Stream watershed,Ribeirão Preto region, SP, Brazil. The HPLC methodused for sample screening was based on herbicideextraction with dichloromethane:isopropanol (9:1, v/v)followed by reversed-phase chromatography (RP-8) withdetection at 220 nm. The presence of herbicides wasconfirmed by GC-MS after ethyl acetate extraction. Atotal of 250 samples collected at different sites fromOctober 1995 to July 1996 were analyzed. Ametrynresidues were detected in 17 samples but almost alwaysat concentrations below those maximum levels recommended by international agencies of environmental control.
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    Autotoxic Compounds from Fresh Alfalfa Leaf Extracts: Identification and Biological Activity (2000)
    Springer
    Journal of chemical ecology 26 (2000), S. 315-327 
    Details
    ISSN: 1573-1561
    Keywords: Alfalfa extract ; autotoxicity ; bioassay ; chlorogenic acid ; salicylic acid ; HPLC ; GC-MS
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Many investigators have attempted to identify the allelochemicals in alfalfa (Medicago sativa), that cause autotoxicity. The autotoxic compounds from fresh alfalfa leaves were separated and quantified, and their biological activity was determined. Chemical separation procedures involved an 80% methanol extract of fresh alfalfa leaves, treatment with activated charcoal, microcrystalline cellulose thin-layer chromatography (MCTLC), and finally separation by Sephadex LH-20 column chromatography. The various fractions were examined further by high-performance liquid chromatography (HPLC). Preliminary identification by HPLC analysis resulted in peaks with retention times close to those of chlorogenic (m/z = 354) and salicylic acid (m/z = 138) standards, and these compounds were confirmed with GC-MS. Several other peaks remain unidentified. Chlorogenic acid occurs in relatively large amounts (0.39 mg/g) in alfalfa aqueous extracts as compared to salicylic acid (0.03 mg/g), and bioassays suggest that chlorogenic acid is involved in alfalfa autotoxicity.
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    HPLC analysis ofArtemisia dracunculus extracts (2000)
    Springer
    Chemistry of natural compounds 36 (2000), S. 144-147 
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    ISSN: 1573-8388
    Keywords: Artemisia dracunculus ; flavonoids ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Flavonoids in extracts ofArtemisia dracunculus L. are studied. The principal component is identified as pinocembrine. Pinocembrine is analyzed quantitatively using an internal standard. The uncertainties in the chromatographic measurements are estimated.
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    Differential Effect of Cd2+ and Ni2+ on Amino Acid Metabolism in Soybean Seedlings (2000)
    Springer
    Biologia plantarum 43 (2000), S. 79-84 
    Details
    ISSN: 1573-8264
    Keywords: alanine aminotransferase ; aspartate aminotransferase ; cysteine ; Glycine max ; heavy metals ; HPLC ; nitrate assimilation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In 10-d-old soybean seedlings, the growth of roots and shoots was significantly inhibited at 50 and 100 μM and more Cd2+, respectively, and by 50 μM or more Ni2+. Although total protein content of roots exposed to 200 μM Cd2+ or Ni2+ was similarly decreased compared to the control, the activity of nitrate reductase was much more inhibited by Cd2+. Ni2+-treatment (200 μM) induced an accumulation of all free amino acids in roots associated with a decrease in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities reflecting the accumulation of both alanine and aspartic acid, respectively. Cd2+-treatment (200 μM) decreased the amount of all free amino acids. In addition, cysteine which is the main amino acid consisting the phytochelatin complexes constituted about 17.5 % of total free amino acids. The activities of both ALT and AST in Cd2+-treated roots were higher than in Ni2+-treated roots suggesting higher conversion of alanine and aspartate to pyruvate and oxaloacetate. Primary leaves excised from either Cd2+ or Ni2+-treated seedlings showed similar pattern of enzyme activities as roots.
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    Suitability of new and old association hosts for the development of selected microgastrine parasitoids of gramineous stemborers (1999)
    Springer
    Entomologia experimentalis et applicata 90 (1999), S. 257-266 
    Details
    ISSN: 1570-7458
    Keywords: host suitability ; acceptance ; biological control ; new associations ; Lepidoptera ; Pyralidae ; New World ; Old World ; stemborers ; Braconidae ; larval parasitoids ; Gramineae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The present study examined the acceptability and suitability of Old World stemborers (Chilo partellus and C. orichalcociliellus) for the development of New World parasitoids (Apanteles deplanatus and A. minator) and New World stemborers (Diatraea saccharalis and D. grandiosella) for the development of Old World parasitoids (Cotesia sesamiae, C. flavipes and C. chilonis). Results revealed that acceptance and suitability were high in old associations. In new associations, parasitoids accepted about 60% of the new association hosts. In addition, 10 out of 17 new associations were successful. Apanteles species appeared to be more physiologically host specific than Cotesia species. For example, two of four new association hosts were accepted by A. deplanatus and only one (D. saccharalis) was partially suitable for progeny development. Among the Cotesia species, Cotesia flavipes appeared to have a wider host range than the two other species. It attacked all hosts offered and successfully parasitized all but one (D. grandiosella). Diatraea saccharalis was accepted and was a suitable host for the development of all parasitoid species tested, whereas D. grandiosella was unsuitable for the development of four out of five parasitoid species tested. No clear pattern was observed as behavioral acceptance did not always agree with the pattern of physiological suitability. Implications of these findings for importation biological control of stemborers are discussed.
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    Dependence on host constituents controlling food acceptance by Manduca sexta larvae (1999)
    Springer
    Entomologia experimentalis et applicata 93 (1999), S. 209-215 
    Details
    ISSN: 1570-7458
    Keywords: host specificity ; experience ; feeding behavior ; stimulants ; deterrents ; Manduca sexta ; Solanaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The host range of Manduca sexta L. (Lepidoptera: Sphingidae) is restricted in nature to plants in the family Solanaceae. However, naive hatchling larvae often accept and continue their development on foliage from a wide spectrum of unrelated plants. In contrast, solanaceous-experienced larvae refuse to feed on other plants. Experiments were designed to explore the role of constituents of various plants in this behavioral phenomenon. Fourth instar larvae reared on solanaceous hosts: tomato (Lycopersicon esculentum Mill.), potato (Solanum tuberosum L.), or tobacco (Nicotiana tabacum L.), and the leguminous non-host cowpea (Vigna sinensis Savi), or on artificial diet based on wheat germ were tested for their acceptance or preference when offered a new diet in both choice and no-choice situations. Under no-choice conditions, acceptability of cowpea foliage was strongly affected by the larval dietary experience. Most larvae reared on solanaceous foliage did not feed, whereas those larvae reared on non-solanaceous food readily accepted cowpea. Moreover, solanaceous foliage, leaf discs and extracts were readily accepted by larvae regardless of their dietary experience. Larvae reared on any of the solanaceous plants strongly preferred cowpea discs that were treated with solanaceous extract, while larvae reared on non-solanaceous diets did not discriminate between treated and control discs. Assays of cowpea extracts as well as acceptance of cowpea discs treated with solanaceous extract indicated a lack of detectable deterrent in cowpea. Our results suggest that the mechanism for induced host specificity in M. sexta larvae involves development of a dependence on solanaceous chemical constituents. We argue that this dependence on host plant chemistry has adaptive significance.
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    Mantle heat flow and geotherms for the main geologic domains in Morocco (1999)
    Springer
    International journal of earth sciences 88 (1999), S. 458-466 
    Details
    ISSN: 1437-3262
    Keywords: Key words Heat flow ; Pn velocity ; Radioactivity ; Geotherm ; Morocco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences
    Notes: Abstract  Thermal and deep seismic soundings data are used to study the dependence between the compressional Pn velocity and the surface heat flow or the temperature at the Moho discontinuity in Morocco. This correlation indicates a significant decrease in Pn velocity where high heat flow and Moho temperature are observed. This result is consistent with respect to other regions of the world. Crustal heat generation models and geotherms are constructed for the major Moroccan geological domains extending from the Precambrian units in the south to the Alpine units in the north. The crustal contribution in surface heat flow is on average 35 mWm–2, with high values of 41–42 mWm–2 in the western and eastern Meseta where Hercynian granite intrusions could enrich the crust in radioactive heat sources. High mantle heat flow values are obtained beneath the Alboran neogene basin (62 mWm–2), the Rif (47 mWm–2), the Middle Atlas (41 mWm–2), and the south Atlantic margin (40 mWm–2) where the crust is thinned by an extensional tectonic regime. Despite their similar formation context, the intra-continental belts of the Middle and the High Atlas show different geothermal field components. A lithospheric heating process in the Middle Atlas could be the result of a Plio-Quaternary basaltic volcanism. Finally, the Precambrian basement of the Anti-Atlas like all the West African shield is a stable domain showing the lowest subsurface temperatures.
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    A new approach for development of rugged sample preparation of metabolites of albendazole in cow milk (1999)
    Springer
    Microchimica acta 130 (1999), S. 155-163 
    Details
    ISSN: 1436-5073
    Keywords: Optimisation ; Taguchi method ; HPLC ; Solid phase extraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Sample preparation is the critical step in analysis of residues in biological samples. The development of a ragged method is time-consuming, because a huge number of parameters must be checked. To reduce the number of experiments Taguchi's method was applied in the sample preparation of metabolites of albendazole. During the experiments 11 controllable and 7 noise factors were investigated. From the influence of controllable and noise factors on recovery and standard deviation, conditions for the sample preparation and recovery could be concluded with high accuracy and reliability.
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    Validation of an HPLC method for the determination of *-dichlorobenzene and naphthalene in mothrepellents (1999)
    Springer
    Accreditation and quality assurance 4 (1999), S. 473-476 
    Details
    ISSN: 1432-0517
    Keywords: Key words Validation ; HPLC ; *-Dichlorobenzene ; Naphthalene ; Mothrepellents.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract  The determination of dichlorobenzene and naphthalene in commercial repellents used in Spain has been validated. This was done using an isocratic regime, to test the reverse -phase HPLC system with acetonitrile: water 65 : 35 (v: v) as the mobile phase, at 20  °C. This technique is proposed for the modular validation of the HPLC system . The results obtained with this method show good agreement with the results provided by the manufacturers of the mothrepellents.
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    Polycyclic Aromatic Hydrocarbons: Need for Assessment of Health Risks in India? Study of An Urban-Industrial Location in India (1999)
    Springer
    Environmental monitoring and assessment 59 (1999), S. 287-319 
    Details
    ISSN: 1573-2959
    Keywords: BaP ; carcinogens ; GC-MS ; HPLC ; polycyclic aromatic hydrocarbons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract This paper reports the PAHs levels in the atmosphere of an urbanised industrial site of India. A high-resolution capillary gas chromatograph with a mass spectrometric detector (HRCGC-MS) and a high performance liquid chromatograph (HPLC) equipped with a fluorescence detector were used for the identification and quantitation of PAHs. The atmospheric levels of PAHs were higher (4.66 ng/m3 yearly average) than most of the concentrations previously reported in the literature. Indian sites were found more contaminated with potently carcinogenic: four and above ringed PAHs. Based on a good correlation between the levels of lead, vanadium, BaP and BghiP, the vehicular emission appears to be a major source of the PAHs. Further, the higher levels of observed PAHs could be attributed to the vertical distribution of the aerosols, the preference of the PAHs for the particulate phase and the greater availability of the substrate in the atmosphere for their sorption. This paper also discusses the need for development of a PAHs monitoring protocol and related health effect studies in developing countries such as India.
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    URL: http://dx.doi.org/10.1023/A:1006169605672
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    The Intermolecular Disulfide Bridge of Human Glial Cell Line-Derived Neurotrophic Factor: Its Selective Reduction and Biological Activity of the Modified Protein (1999)
    Springer
    The protein journal 18 (1999), S. 585-593 
    Details
    ISSN: 1573-4943
    Keywords: Glial cell line-derived neurotrophic factor ; partial reduction ; HPLC ; chemical modification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Recombinant human glial cell line-derived neurotrophic factor has been implicated to have therapeutic potential in the treatment of neurodegenerative diseases. The mature protein is a single polypeptide of 134 amino acid residues and functions as a disulfide-linked dimer. Reduction of the protein with dithiothreitol at pH 7.0 and in the absence of denaturant showed that the single intermolecular cystine bridge was reduced preferentially. Direct alkylation of the generated free sulfhydryl group using iodoacetamide or iodoacetate without denaturant was incomplete. Unfolding the protein in 6 M guanidine hydrochloride prior to the modification showed rapid disulfide scrambling. However, the sulfhydryl-modifying reagent N-ethylmaleimide was able to label quantitatively the free cysteinyl residue in the absence of any added chaotropic agent. By a combination of peptide mapping, Edman degradation, and mass spectrometric analysis, the labeled residue was identified to be Cys101, hence verifying the location of the intermolecular disulfide bond. The modified protein behaved as a noncovalent dimer when chromatographed through a Superdex 75 column under nondenaturing conditions and was comparable in biological activity to an unmodified control sample. The results therefore indicate that the intermolecular disulfide bridge of the protein is not essential for its biological function.
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    URL: http://dx.doi.org/10.1023/A:1020607501910
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    Biosynthesis and Characterization of 4-Fluorotryptophan-Labeled Escherichia coli Arginyl-tRNA Synthetase (1999)
    Springer
    The protein journal 18 (1999), S. 187-192 
    Details
    ISSN: 1573-4943
    Keywords: Arginyl-tRNA synthetase ; 4-fluorotryptophan ; 19F NMR ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Escherichia coli 4-fluorotryptophan-substituted arginyl-tRNA synthetase was biosynthetically prepared and purified from a tryptophan auxotroph which could overproduce this enzyme. A method was developed to separate 4-fluorotryptophan from tryptophan and to determine accurately their contents in the 4-fluorotryptophan-containing proteins. It was confirmed that more than 95% of the tryptophan residues in the purified 4-fluorotryptophan-substituted arginyl-tRNA synthetase were replaced by 4-fluorotryptophan. Studies on the effect of the 4-fluorotryptophan replacement on properties of the enzyme showed that, when compared with the native enzyme, both the specific activity and the first-order rate constant of the fluorinated enzyme decreased by approximately 20% with just slightly higher K m values. CD studies, however, did not reveal any difference between the secondary structure of the native and fluorinated enzymes. In addition, thermal unfolding studies showed that the 4-fluorotryptophan replacement did not significantly affect the thermal stability of the enzyme. We may conclude that the substitution of 4-fluorotryptophan in arginyl-tRNA synthetase had no substantial effect on the structure and function of the enzyme. Finally, a preliminary study of 19F nuclear magnetic resonance spectroscopy of the fluorinated enzyme has shown promising prospect for further investigation of its structure and function with NMR.
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    URL: http://dx.doi.org/10.1023/A:1020675922382
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    Spectrum of resistance to root-knot nematodes and inheritance of heat-stable resistance in in pepper (Capsicum annuum L.) (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 496-502 
    Details
    ISSN: 1432-2242
    Keywords: Key words Capsicum annuum L. ; Solanaceae ; Meloidogyne spp. ; Nematode resistance ; Dominant Me3 gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Capsicum annuum L. has resistance to root-knot nematodes (RKN) (Meloidogyne spp.), severe polyphagous pests that occur world-wide. Several single dominant genes confer this resistance. Some are highly specific, whereas others are effective against a wide range of species. The spectrum of resistance to eight clonal RKN populations of the major Meloidogyne species, M. arenaria (2 populations), M. incognita (2 populations), M. javanica (1 population), and M. hapla (3 populations) was studied using eight lines of Capsicum annuum. Host susceptibility was determined by counting the egg masses (EM) on the roots. Plants were classified into resistant (R; EM ≤ 5) or susceptible (H; EM 〉5) classes. The french cultivar Doux Long des Landes was susceptible to all nematodes tested. The other seven pepper lines were highly resistant to M. arenaria, M. javanica and one population of M. hapla. Variability in resistance was observed for the other two populations of M. hapla. Only lines PM687, PM217, Criollo de Morelos 334 and Yolo NR were resistant to M. incognita. To investigate the genetic basis of resistance in the highly resistant line PM687, the resistance of two progenies was tested with the two populations of M. incognita: 118 doubled-haploid (DH) lines obtained by androgenesis from F1 hybrids of the cross between PM687 and the susceptible cultivar Yolo Wonder, and 163 F2 progenies. For both nematodes populations, the segregation patterns 69 R / 49 S for DH lines and 163 R / 45 S for F2 progenies were obtained at 22°C and at high temperatures (32°C and 42°C). The presence of a single dominant gene that totally prevented multiplication of M. incognita was thus confirmed and its stability at high temperature was demonstrated. This study confirmed the value of C. annuum as a source of complete spectrum resistance to the major RKN.
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    URL: http://dx.doi.org/10.1007/s001220051262
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    AFLP analysis of genetic relationships among the cultivated eggplant, Solanummelongena L., and wild relatives (Solanaceae) (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 626-633 
    Details
    ISSN: 1432-2242
    Keywords: Key words DNA ; AFLP ; Solanaceae ; Solanum ; Eggplant ; Phenetic analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The AFLP technique of DNA analysis was evaluated as a tool for assessing genetic relationships among the cultivated eggplant, S. melongena, and related species [Solanum L. subgenus Leptostemonum (Dunal) Bitter, section Melongena (Mill.) Dunal, series Incaniformia Bitter]. Genetic distances based on the AFLP data were estimated for 49 samples of 36 distinct accessions. Phenetic trees were constructed using Jaccard’s coefficient and UPGMA, and other clustering methods: they all had very high co-phenetic correlation values, and were found to be consistent with previous trees based on other data types, in particular ITS-1 sequences, isozymes and morphology, carried out on the same accessions. These results indicated that the AFLP technique is both an efficient and effective tool for determining genetic relationships among species of Solanum. A new classification is proposed for series Incaniformia.
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    URL: http://dx.doi.org/10.1007/s001220051277
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    AFLP analysis of genetic relationships in the tribe Datureae (Solanaceae) (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 634-641 
    Details
    ISSN: 1432-2242
    Keywords: Key words DNA ; AFLP ; Solanaceae ; Datura ; Brugmansia ; phenetic analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The AFLP technique was evaluated as a tool for assessing species relationships within the tribe Datureae and genetic distances were estimated for 47 accessions of over 12 species. The phenetic trees from various analyses of the AFLP data gave very high co-phenetic correlation values, and were found to be consistent with previous trees based on the analysis of different data types, in particular ITS-1 sequences, isozymes and morphology, carried out on the same accessions. These results indicated that the AFLP technique is both an efficient and effective tool for determining genetic relationships among taxa in the Solanaceae. A new classification is proposed for the tribe Datureae, which maintains the arborescent species as a separate genus, Brugmansia, and recognises three sections within the genus Datura; Stramonium, Dutra and Ceratocaulis. D. discolor, previously placed in section Dutra, was found to be intermediate between sections Dutra and Stramonium.
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    URL: http://dx.doi.org/10.1007/s001220051278
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    Residues of free and bound maleic hydrazide in potato tubers (1999)
    Springer
    Potato research 42 (1999), S. 89-93 
    Details
    ISSN: 1871-4528
    Keywords: analysis ; HPLC ; β-glucoside ; 1,2-dihydro-3,6-pyridazinedione ; Solanum tuberosum L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The method of Vadukul (1991) for determining maleic hydrazide (MH) was modified and gave recoveries of free MH of 89%±4%. The values recorded on individual tubers ranged from 2–14 mg kg−1. Maleic hydrazide was evenly distributed throughout the tuber (peel, outer and inner flesh) but concentration increased slightly as tuber size increased. The concentration of free MH decreased from 7 to 3 mg kg−1 over the storage period of 5 1/2 months. Acid hydrolysis released substantial amounts of MH particularly from older potatoes (13 mg kg−1) compared with 6 mg kg−1 from new potatoes, implying that free MH is gradually converted to a bound form with time after treatment. No evidence was found for the presence of a β-glucoside of MH.
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    The recovery of maleic hydrazide from processed potato products (1999)
    Springer
    Potato research 42 (1999), S. 95-99 
    Details
    ISSN: 1871-4528
    Keywords: analysis ; HPLC ; filter paper model ; Diels-Alder reaction ; 1,2-dihydro-3,6-pyridazinedione ; Solanum tuberosum L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A method used for estimating extractable maleic hydrazide (MH) concentrations in fresh potato material, concentration range 5–17 mg kg−1, was found not to be suitable for processed potato products (10–33% recoveries) although, boiling potato pieces enhanced recovery by 20%. Each step of the determination was examined and a modified procedure developed with particular emphasis on the extraction of MH from the dried potato matrix, and the quality of the HPLC column used. Potato slices and model systems based on filter papers plus additives were used. Recoveries from fried potato slices were 74±6%. Based on the effect of glucose in reducing extractable MH recoveries, it is suggested that the remainder of the MH (20–25%) is converted into a conjugated structure on reaction with dehydrated sugar (Diels-Alder reaction).
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    Detection of laxative abuse by urine analysis with HPLC and diode array detection (1999)
    Springer
    Pharmacy world & science 21 (1999), S. 40-43 
    Details
    ISSN: 1573-739X
    Keywords: Laxative abuse ; Factitial diarrhea ; Chronic diarrhea ; Urine analysis ; Bisacodyl ; Bisoxatin ; Phenolphthalein ; Emodine ; Aloe‐emodine ; Rheine ; Danthron ; Picosulphate ; HPLC ; Diode array
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract A simple method is proposed for analysis of stimulant laxatives and metabolites of laxatives in urine. All stimulant laxatives commercially available in Germany, Begium and the Netherlands, the diphenylmethane derivatives and the anthraquinones, were included. Chromatography was performed with a standardized isocratic HPLC system with diode array detection ('STIP'), which is commonly used in the Netherlands for toxicological screening. The method was validated by ingestion of a normal dose of the laxatives by human volunteers. In all cases the expected laxative metabolite could be detected in urine twelve hours after intake. Also urine samples of patients, suspected of laxative abuse, were analyzed.
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    URL: http://dx.doi.org/10.1023/A:1008647424809
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    Isolation, Characterization, and Stability of Positional Isomers of Mono-PEGylated Salmon Calcitonins (1999)
    Springer
    Pharmaceutical research 16 (1999), S. 813-818 
    Details
    ISSN: 1573-904X
    Keywords: calcitonin ; polyethylene glycol ; PEGylation ; peptide ; tryptic digestion ; stability ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. To separate and characterize the different positional isomers of mono-PEGylated salmon calcitonins (mono-PEG-sCTs) and to evaluate the effects of the PEGylation site on the stability of different mono-PEG-sCTs in rat kidney homogenate. Methods. Mono-PEG-sCTs were prepared using succinimidyl carbonate monomethoxy polyethylene glycol (5,000 Da) and separated by gel-filtration HPLC followed by reversed-phase HPLC. To characterize PEGylated sCTs, matrix-assisted laser desorption ionization time of flight mass spectrometry (M ALDI-TOF MS) and reversed-phase HPLC of the trypsin digested samples were performed. Mono-PEG-sCTs and sCT in rat kidney homogenates were measured by column-switching reversed-phase HPLC with on-line detection of the radioiodinated samples using a flow-through radioisotope detector. Results. Three different mono-PEGylated sCTs were separated by reversed-phase gradient HPLC. From the MALDI-TOF MS analysis, the average molecular weight of mono-PEG-sCTs was confirmed as around 8650 Da. The presence of PEG moiety in the mono-PEG-sCTs was also manifested by the fact that the distance between two adjacent mass spectum lines was 44 Da which corresponds to PEG monomer unit. Tryptic digestion analysis demonstrated that these mono-PEG-sCTs are 3 positional isomers of N-terminus, Lys18- and Lys11-residue modified mono-PEGylated sCTs. The degradation half-life of these 3 positional isomers in rat kidney homogenates significantly increased in order of the N-terminus (125.5 min), Lys11- (157.3 min), and Lysl8-residue modified mono-PEGylated sCT (281.5 min) over the native sCT (4.8 min). Conclusions. Three positional isomers of mono-PEGylated sCTs were purified and characterized. Of these, the resistance to proteolytic degradation was highest for the Lysl8-residue modified mono-PEG-sCT. These studies demonstrate that the in vivo stability of PEGylated sCTs is highly dependent on the site of PEG molecule attachment.
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    URL: http://dx.doi.org/10.1023/A:1018861616465
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    Evaluation of the Pharmacokinetic Features and Tissue Distribution of the Potent Nonnucleoside Inhibitor of HIV-1 Reverse Transcriptase, N-[2-(2-fluorophenethyl)]-N′-[2-(5-bromopyridyl)]-thiourea (HI-240) with an Analytical HPLC Method (1999)
    Springer
    Pharmaceutical research 16 (1999), S. 1226-1232 
    Details
    ISSN: 1573-904X
    Keywords: HI-240 ; nonnucleoside inhibitor ; pharmacokinetics ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. The purpose of the present study was to examine the pharmacokinetic features and tissue distribution of N-[2-(2-fluorophenethyl)]-N′-[2-(5-bromopyridyl)]-thiourea (HI-240), a novel non-nucleoside inhibitor of HIV reverse transcriptase with potent anti-viral activity against AZT-sensitive as well as multidrug-resistant HIV-1 strains. Methods. A sensitive and accurate high performance liquid chromatography (HPLC)-based quantitative detection method was established to measure concentrations of HI-240 in pharmacokinetic studies. The plasma concentration-time data were modeled by using the WinNonlin program to estimate the pharmacokinetic parameter values. Results. HI-240 had an elimination half-life of 78.3 ± 2.0 min after i.v. administration and 196.8 ± 3.1 min after i.p. administration. The systemic clearance of HI-240 was 2194 ± 61 ml/h/kg after i.v. administration and 9339 ± 1160 ml/h/kg after i.p. administration. Following i.v. injection, HI-240 rapidly distributed to and accumulated in multiple tissues with particularly high accumulation in adipose tissue, adrenal gland, and uterus+ovary. The concentration of HI-240 in brain tissue was comparable to that in the plasma, indicating that HI-240 easily crosses the blood-brain-barrier. Following i.p. injection, HI-240 was rapidly absorbed with a t1/2ka and a tmax values of less than 10 min. Following oral administration, HI-240 was absorbed with a t1/2ka of 4.2 ±1.1 min and a tmax of 95.1 ± 25.1 min. The intraperitoneal bioavailability was estimated at 23.5%, while the oral bioavailability was only 1%. Conclusions. The HPLC-based accurate and precise analytical detection method and pilot pharmacokinetic studies described herein provide the basis for advanced preclinical pharmacodynamic studies of HI-240. The ability of HI-240 to distribute rapidly and extensively into extravascular compartments and easily cross the blood-brain barrier represent significant pharmacokinetic advantages over AZT.
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    URL: http://dx.doi.org/10.1023/A:1014814313681
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    Pharmacokinetic Features and Metabolism of Calphostin C, A Naturally Occurring Perylenequinone with Antileukemic Activity (1999)
    Springer
    Pharmaceutical research 16 (1999), S. 1003-1009 
    Details
    ISSN: 1573-904X
    Keywords: pharmacokinetics ; Calphostin C ; HPLC ; perylenequinone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Purpose. To examine the pharmacokinetic features and metabolism of calphostin C, a naturally occurring perylenequinone with potent antileukemic activity. Methods. HPLC-based quantitative detection methods were used to measure calphostin C levels in lysates of leukemic cells and in plasma of mice treated with calphostin C. The plasma concentration-time data were analyzed using the WinNonlin program. In vitro esterases and a microsome P450 preparation in conjunction with a LC-MS(API-EI) system were used to study the metabolism of calphostin C. Results. An intracellular exposure level (AUC0−6h) of 257 μM·h was achieved after in vitro treatment of NALM-6 cells with calphostin C at a 5 μM final concentration in culture medium. After intraperitoneal (i.p.) injection of a 40 mg/kg nontoxic bolus dose of calphostin C, the estimated Cmax was 2.9 μM, which is higher than the effective in vitro concentration of calphostin C against leukemic cells. Drug absorption after i.p. administration was rapid with an absorption half-life of 24.2 min and the estimated tmax was 63.0 min. Calphostin C was cleared with an elimination half-life of 91.3 min. An inactive and smaller metabolite (calphostin B) was detected in plasma of calphostin C-treated mice with a tmax of 41.3 min. Esterase (but not P450) treatment of calphostin C in vitro yielded an inactive metabolite (calphostin B) of the same size and elution profile. Conclusions. Target plasma calphostin C concentrations of potent antileukemic activity can be reached in mice at nontoxic dose levels. This pilot pharmacokinetic study of calphostin C combined with the availability of the described quantitative HPLC method for its detection in cells and plasma provide the basis for future preclinical evaluation of calphostin C and its potential as an anti-leukemic drug.
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    URL: http://dx.doi.org/10.1023/A:1018923430094
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    25S-18S rDNA intergenic spacer ofNicotiana sylvestris (Solanaceae): Primary and secondary structure analysis (1999)
    Springer
    Plant systematics and evolution 218 (1999), S. 89-97 
    Details
    ISSN: 1615-6110
    Keywords: Solanaceae ; Nicotiana ; rDNA ; subrepeats ; molecular evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The complete nucleotide sequence (3701 bp) of the intergenic spacer region between the 3′-end of 25S rRNA gene and the 5′-end of 18S rRNA gene (IGS) is described forNicotiana sylvestris. The IGS ofN. sylvestris consists of seven structural regions (SR) two of which are composed of subrepeats (SR II: three and one half copies of the 76 bp D-subrepeat and numerous short C-subrepeats; SR IV: five copies of the 142/136 bp A-subrepeat). A presumptive rRNA processing signal (CTTTT) is located at the beginning of SR I. An identical sequence is also present in each copy of the A-subrepeat. Each D-subrepeat in SR IIa contains at the 5′-end the conserved sequence (GAGGTTTTT) which may be involved in transcription termination. SR III (TG-rich) and SR IV (AT-rich) in the central part of IGS contain several potential replication origin sites. The transcription initiation site (TATATAAGGGGGG) is located at the border of SR IV and V. No spacer promoters have been found. SR V contains four copies of a CATGG motif that is also present in SR II, VI and VII. At the 5′-end of SR VII there are three copies of the 66 bp highly divergent B-subrepeat. Presumptive pre-rRNA processing signals are also present here. Possible hair-pin structures are described for SR V, VI and VII.
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    Collapse of species boundaries in the wild potatoSolanum brevicaule complex (Solanaceae, S. sect.Petota): Molecular data (1999)
    Springer
    Plant systematics and evolution 214 (1999), S. 103-130 
    Details
    ISSN: 1615-6110
    Keywords: Solanaceae ; sect.Petota ; Solanum brevicaule ; Domestication ; hybridization ; potatoes ; systematics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract TheSolanum brevicaule complex is a group of morphologically very similar wild and cultivated potato taxa (Solanum sect.Petota). This study uses single to low-copy nuclear RFLPs and RAPDs to investigate their species boundaries and relationships. Cladistic analyses of both data sets are largely concordant with each other and with a recently published phenetic analyses of the same accessions using morphology. All three data sets separate members of the complex into populations from Peru and immediately adjacent northwestern Bolivia, including most cultivated species accessions, and populations from northwestern Bolivia to Argentina. The molecular results suggest that the complex is paraphyletic as currently circumscribed. Many species of theS. brevicaule complex should be relegated to synonymy.
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    Random mutagenesis in the large extrinsic loop E and transmembrane α-helix VI of the CP 47 protein of Photosystem II (1999)
    Springer
    Plant molecular biology 39 (1999), S. 381-386 
    Details
    ISSN: 1573-5028
    Keywords: Photosystem II ; CP 47 ; random mutagenesis ; XL-1 Red ; mutator strains
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The intrinsic chlorophyll-protein CP 47 is a component of Photosystem II which functions in both light-harvesting and oxygen evolution. Using the Escherichia coli mutator strain XL-1 Red, we introduced mutations at 14 sites in the large extrinsic loop E of CP 47 and its adjacent transmembrane α-helix VI. Four mutant cell lines were recovered in which the histidyl residues 455H, 466H and 469H were altered. The cell lines H455T, H455Y, H469Y, and the double mutant F432L,H466R exhibited phenotypes that supported the identification of the histidyl residues 455H, 466H and 469H as chlorophyll ligands. Four additional mutant cell lines were recovered which contained mutations at positions 448R in the large extrinsic loop of CP 47. These mutants, R448K, R448Q, R448S, and R448W, exhibited variable phenotypes ranging from moderate alteration of photoautotrophic growth and oxygen evolution rates to a complete inhibition of these parameters. Those mutants exhibiting photoautotrophic growth and oxygen evolution capability under standard conditions were unable to grow photoautotrophically or evolve oxygen when grown at low chloride concentrations. Finally, a mutant cell line exhibiting a substitution at position 342G was recovered. The mutant G342D exhibited moderate alterations of photoautotrophic growth and oxygen evolution. In addition to these alterations, mutants were recovered in which deletions and insertions (leading to frame shifts) and stop codons were introduced. These mutants uniformly lacked the ability to either grow photoautotrophically or evolve oxygen.
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    URL: http://dx.doi.org/10.1023/A:1006199901167
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    Disruption of specific flavonoid genes enhances the accumulation of flavonoid enzymes and end-products in Arabidopsis seedlings (1999)
    Springer
    Plant molecular biology 40 (1999), S. 45-54 
    Details
    ISSN: 1573-5028
    Keywords: antibodies ; Arabidopsis ; flavonoid biosynthesis ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Polyclonal antibodies were developed against the flavonoid biosynthetic enzymes, CHS, CHI, F3H, FLS, and LDOX from Arabidopsis thaliana. These antibodies were used to perform the first detailed analysis of coordinate expression of flavonoid metabolism at the protein level. The pattern of flavonoid enzyme expression over the course of seedling development was consistent with previous studies indicating that chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), and flavonol synthase (FLS) are encoded by ‘early’ genes while leucoanthocyanidin dioxygenase (LDOX) is encoded by a ‘late’ gene. This sequential expression may underlie the variations in flavonoid end-products produced during this developmental stage, as determined by HPLC analysis, which includes a shift in the ratio of the flavonols, quercetin and kaempferol. Moreover, immunoblot and HPLC analyses revealed that several transparent testa lines blocked at intermediate steps of the flavonoid pathway actually accumulated higher levels of specific flavonoid enzymes and end-products. These results suggest that specific intermediates may act as inducers of flavonoid metabolism.
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    URL: http://dx.doi.org/10.1023/A:1026414301100
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    Protoplast and leaf explant culture of Lycopersicon cheesmanii and salt tolerance of protoplast-derived calli (1999)
    Springer
    Plant cell, tissue and organ culture 56 (1999), S. 9-16 
    Details
    ISSN: 1573-5044
    Keywords: in vitro shoot regeneration ; micropropagation ; salinity ; Solanaceae ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In vitro manipulation of Lycopersicon cheesmanii was attempted in order to evaluate its potential to improve salt tolerance of L. esculentum following somatic hybridisation of these two species. The main study concerned three populations of the typicum form (LCS) used for tissue culture and protoplast studies. The minor halophytic form (LCT) was used only for protoplast experiments. Mother plants were sown and propagated in vitro. Shoots (2-7) were obtained from explants (leaflets, petioles, cotyledons), through calli, when 3-4 mg l−1 BA, kinetin or 2iP were used in association with auxin. IAA (0.2 and 1 mg l−1 was the most efficient, while 2,4-D or NAA were sometimes inhibitory according to genotype and explant. Leaf protoplasts (2-5 × 107/g of fresh weight) were obtained from LCS and LCT. For LCT, protoplast division declined after five subcultures of the plantlets that had been established from seeds. Protoplast-derived calli were obtained from all populations (maximum PE 30% for LCS). However, they failed to regenerate shoots. Protoplast-derived calli involved in salt tolerance tests showed that LCS17 line was the most tolerant.
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    URL: http://dx.doi.org/10.1023/A:1006253000482
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    The effect of copper on chlorophyll organization during greening of barley leaves (1999)
    Springer
    Photosynthesis research 62 (1999), S. 165-174 
    Details
    ISSN: 1573-5079
    Keywords: chlorophyll fluorescence ; oxygen evolution ; photoacoustic ; photodamage ; Photosystem II
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of copper on chlorophyll organization and function during greening of barley was examined, using chlorophyll fluorescence and photoacoustic techniques. Copper was found to inhibit pigment accumulation and to retard chlorophyll integration into the photosystems, as evident from low temperature (77 K) fluorescence spectra. Resolution of the minimal fluorescence (F0) into active and inactive parts, indicated a higher inactive fraction with copper treatment. This was attributed to chlorophyll molecules which failed to integrate normally, a conclusion supported by the longer fluorescence lifetime observed in copper treated plants. A lower ratio of chlorophyll a to b and fluorescence induction transients, showing accelerated Photosystem II closure, both indicate that copper treatment resulted in a larger light-harvesting antenna. Another effect of copper treatment was the suppression of oxygen evolution, indicating a decrease in photosynthetic capacity. We suggest that the non-integrated chlorophyll fraction sensitizes photodamage in the membrane, contributing to disruption of electron flow and pigment accumulation.
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    URL: http://dx.doi.org/10.1023/A:1006397714430
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    Dark reoxidation of the plastoquinone-pool is mediated by the low-potential form of cytochrome b-559 in spinach thylakoids (1999)
    Springer
    Photosynthesis research 62 (1999), S. 273-279 
    Details
    ISSN: 1573-5079
    Keywords: plastoquinone ; cytochrome b-559 ; thylakoid membrane ; Photosystem II ; chlororespiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have found that in isolated spinach thylakoids, plastoquinone-pool (PQ-pool), after its photoreduction, undergoes dark-reoxidation with the half-time of τ1/2 = 43 ± 3 s. To explain the observed rates of PQ-pool reoxidation, a nonenzymatic plastoquinol (PQH2) autoxidation under molecular oxygen and an enzymatic oxidation by the low-potential form of cytochrome b-559 (cyt. b-559LP), as the postulated PQ-oxidase in chlororespiration, were investigated. It was found that the autoxidation rate of PQH2 in organic solvents and liposomes was too low to account for the observed oxidation rate of PQH2 in thylakoids. The rate of cyt. b-559LP autoxidation in isolated Photosystem II was found to be similar (τ1/2 = 26 ± 5 s) to that of the PQ-pool. This suggests that the LP form of cyt. b-559 is probably responsible for the PQ-oxidase activity observed during chlororespiration.
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    URL: http://dx.doi.org/10.1023/A:1006374319191
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    Effects of anaerobiosis as probed by the polyphasic chlorophyll a fluorescence rise kinetic in pea (Pisum sativum L.) (1999)
    Springer
    Photosynthesis research 62 (1999), S. 67-83 
    Details
    ISSN: 1573-5079
    Keywords: anaerobiosis ; chlorophyll fluorescence ; chlororespiration ; fluorescence induction ; Photosystem II ; Pisum sativum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We analysed the changes of the chlorophyll (Chl) a fluorescence rise kinetic (from 50 μs to 1 s) that occur when leaves or chloroplasts of pea ( Pisum sativum L.) are incubated under anaerobic conditions in the dark. In control leaves, Chl a fluorescence followed a typical O-J-I-P polyphasic rise [Strasser et al. (1995) Photochem Photobiol 61: 32–42]. Anaerobiosis modified the shape of the transient with the main effect being a time-dependent increase in the fluorescence yield at the J-step (2 ms). Upon prolongation of the anaerobic treatment (〉 60 min), the O-J-I-P fluorescence rise was eventually transformed to an O-J (J = P) rise. A similar transformation was observed when pea leaves were treated with DCMU or sodium dithionite. Anaerobiosis resulted in a 10–20% reduction in the maximum quantum yield of the primary photochemistry of Photosystem II, as measured by the ratio of the maximal values of variable and total fluorescence (FV/FM). When the leaves were returned to the air in the dark, the shape of the fluorescence transient showed a time-dependent recovery from the anaerobiosis-induced change. The original O-J-I-P shape could also be restored by illuminating the anaerobically treated samples with far-red light but not with blue or white light. Osmotically broken chloroplasts displayed under anaerobic conditions fluorescence transients similar to those observed in anaerobically treated leaves, but only when they were incubated in a medium comprising reduced pyridine nucleotides (NADPH or NADH). As in intact leaves, illumination of the anaerobically treated chloroplasts by far-red light restored the original O-J-I-P transient, although only in the presence of methyl viologen. The results provide additional evidence for the existence of a chlororespiratory pathway in higher plant cells. Furthermore, they suggest that the J-level of the fluorescence transient is strongly determined by the redox state of the electron carriers at the PS II acceptor side.
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    URL: http://dx.doi.org/10.1023/A:1006321126009
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    Retrolyc1-1, a member of the Tntl retrotransposon super-family in the Lycopersicon peruvianum genome (1999)
    Springer
    Genetica 107 (1999), S. 65-72 
    Details
    ISSN: 1573-6857
    Keywords: Lycopersicon ; Retrolyc1 ; retrotransposon ; Solanaceae ; Tntl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Retrotransposons are ubiquitous mobile genetic elements that transpose through an RNA intermediate. One of the best known plant retrotransposon, Tnt1, was isolated from tobacco and showed an extensive distribution in the Nicotiana genus. We investigated the presence of related sequences in the Lycopersicon genus, another member of the Solanaceae family. Hybridization experiments performed using Tnt1 probes indicated that homologous sequences were present in all Lycopersicon species, indicating that these Tnt1-related sequences, that we named Retrolyc1, are distributed throughout the Lycopersicon genus. Different distribution patterns were detected between species, demonstrating a potential use of Retrolyc1 elements as molecular markers. An incomplete Retrolyc1 sequence, that we named Retrolyc1-1, was isolated from an L. peruvianum genomic library. Retrolyc1-1 shows extensive homology with Tnt1 sequences except in the LTR U3 region. Since this region is known to be involved in the control of transcription, this strongly suggests the existence of different patterns of regulation for Tnt1 and Retrolyc1 elements. The study of these two elements within the Solanaceae family may provide interesting models for retrotransposon evolution within this group and transmission in host genomes.
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    URL: http://dx.doi.org/10.1023/A:1004028002883
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    In vitro callus induction and regeneration studies in Withania somnifera (1999)
    Springer
    Plant cell, tissue and organ culture 57 (1999), S. 23-27 
    Details
    ISSN: 1573-5044
    Keywords: Solanaceae ; tissue culture ; Ashwagandha ; medicinal plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Callus cultures were initiated from axillary leaves, axillary shoots, hypocotyls, and root segments on Murashige and Skoog (MS) (1962) medium supplemented with 2,4-D (2 mg l−1) and KN (0.2 mg l−1). Shoots differentiated best from axillary shoot base callus on MS medium containing BA (2 mg l−1). Regenerated shoots rooted best on MS medium containing IBA (2 mg l−1) alone, and IBA (2 mg l−1) with IAA (2 mg l−1). Plantlets were transferred to pots containing sand and soil mixture, acclimatized in a culture room and afterwards transferred to the glasshouse.
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    Chemical potential of Aphelandra sp. cell cultures (1999)
    Springer
    Plant cell, tissue and organ culture 58 (1999), S. 133-140 
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    ISSN: 1573-5044
    Keywords: callus culture ; ESI-MS ; HPLC ; polyamines ; secondary metabolites ; verbascoside
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Six different callus lines and three different suspension culture lines were established from plants of two Aphelandra species (Acanthaceae). All established lines were analyzed for secondary metabolite accumulation. A discrepancy between secondary metabolites accumulated in the plants and in the cell cultures could be observed. All established Aphelandrasp. cell cultures produced verbascoside (acteoside) as the major extractable metabolite. Time course experiments were carried out to investigate the relationship between cell growth and verbascoside production. In the present study it was shown that verbascoside accumulation was growth dependent and positively related to the presence of 2,4-D in the medium. The conditions in which verbascoside represents ca. 18% of cell culture weight have been defined. Free polyamines were detected in the cell culture lines cultivated in MS liquid medium (cysteine 10 mg l-1, thiamine 1 mg l-1, 2,4-D 1 mg l-1, kinetin 0.2 mg l-1 and sucrose 30 g l-1). Putrescine and spermidine accumulated within 8 days to a maximum of 8.4 μmol g-1 of dry wt and 2.6 μmol g-1 of dry wt respectively and thereafter their concentration decreased rapidly. There was no evidence for the presence of spermine or any other type of free or conjugated polyamines in the tested cell culture lines.
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    Isolation and characterization of chloroplast Photosystem II antenna of spinach by reversed-phase liquid chromatography (1999)
    Springer
    Photosynthesis research 61 (1999), S. 281-290 
    Details
    ISSN: 1573-5079
    Keywords: antenna system ; chlorophyll–proteins ; HPLC ; LHC II ; Photosystem II ; spinach
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The protein components of the Photosystem II antenna system, isolated from spinach thylakoids, have been resolved by reversed-phase high performance liquid chromatography (RP-HPLC) using a butyl-silica stationary phase packed either into analytical or semi-preparative columns. Peak identification has been accomplished by a combination of various SDS–PAGE systems employing either Comassie (or silver) staining or immunological detection using polyclonal antibodies raised against LHC II and against CP29, CP26 and CP24 proteins and by aminoacid microsequence. Moreover, peak identification is consistent with the molecular masses determined by Electrospray Ionization Mass Spectrometry (HPLC-ESI-MS). The developed RP-HPLC method allows the resolution of all the protein components of the Photosystem II major Light Harvesting Complex (LHC II) and minor PS II antenna complex (CP24, CP26 and CP29) from grana membranes (BBY) and estimation of their relative stoichiometry in natural and stressed conditions, avoiding the expensive and time consuming separation procedure by sucrose-gradient ultracentrifugation and isoelectrofocusing.
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    Hg2+, Cu2+, and Pb2+ -induced changes in Photosystem II photochemical yield and energy storage in isolated thylakoid membranes: A study using simultaneous fluorescence and photoacoustic measurements (1999)
    Springer
    Photosynthesis research 59 (1999), S. 167-174 
    Details
    ISSN: 1573-5079
    Keywords: cyclic electron transport ; fluorescence ; metal ions ; Photosystem II ; thermal dissipation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Simultaneous fluorescence and photoacoustic measurements have been used to study the effects of metal ions (copper, lead, and mercury) during dark incubation of thylakoid membranes. The values of the chlorophyll fluorescence parameters Fo (initial fluorescence yield with the reaction centers in the open state), Fm (maximal fluorescence yield), Ft (steady state fluorescence yield) and the calculated parameters, Φo (maximal quantum yield of Photosystem II photochemistry) and Φt (actual quantum yield of Photosystem II photochemistry), strongly decreased in the presence of the metal ions coinciding with an increase in the non-photochemical deexcitation rate constant k(N). It was observed that photosynthetic energy storage measured by photoacoustic spectroscopy also decreased but a large portion of energy storage remained unaffected even at the highest metal ion concentrations used. A maximal inhibition of photosyntheti c energy storage of 80% and 50% was obtained with Hg2+ and Cu2+-treated thylakoids, respectively, while energy storage was insensitive to Pb2+. The results are consistent with the known predominant inhibition of the donor side of Photosystem II by the metal ions. The insensitive portion of energy storage is attributed to the possible recurrence of cyclic electron transport around Photosystem II that would depend on the extent of inhibition produced on the acceptor side by the metal ion used.
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    URL: http://dx.doi.org/10.1023/A:1006194621553
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    Structural predictions on the 33 kDa extrinsic protein associated to the oxygen evolving complex of photosynthetic organisms (1999)
    Springer
    Photosynthesis research 61 (1999), S. 11-21 
    Details
    ISSN: 1573-5079
    Keywords: manganese-stabilizing protein ; oxygen evolution ; Photosystem II ; protein structure prediction ; PsbO protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Modern computational methods for protein structure prediction have been used to study the structure of the 33 kDa extrinsic membrane protein, associated to the oxygen evolving complex of photosynthetic organisms. A multiple alignment of 14 sequences of this protein from cyanobacteria, algae and plants is presented. The alignment allows the identification of fully conserved residues and the recognition of one deletion and one insertion present in the plant sequences but not in cyanobacteria. A tree of similarity, deduced from pair-wise comparison and cluster analysis of the sequences, is also presented. The alignment and the consensus sequence derived are used for prediction the secondary structure of the protein. This prediction indicates that it is a mainly-beta protein (25–38% of β-strands) with no more than 4% of α-helix. Fold recognition by threading is applied to obtain a topological 2D model of the protein. In this model the secondary structure elements are located, including several highly conserved loops. Some of these conserved loops are suggested to be important for the binding of the 33 kDa protein to Photosystem II and for the stability of the manganese cluster. These structural predictions are in good agreement with experimental data reported by several authors.
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    URL: http://dx.doi.org/10.1023/A:1006265816104
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    Correlations between chlorophyll fluorescence quenching parameters and photosynthesis in a segregating Lycopersicon esculentum × L. peruvianum population as measured under constant conditions (1999)
    Springer
    Photosynthesis research 61 (1999), S. 145-156 
    Details
    ISSN: 1573-5079
    Keywords: chilling ; Photosystem II ; quantum efficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chl fluorescence of mature leaves in low-temperature treated plants was studied under identical measuring conditions in a segregating population of the F3 offspring of a cross between a chilling-tolerant and a chilling-sensitive tomato species. Through recombination of genes involved in photosynthesis, the population revealed a wide, continuous variability of photosynthetic capacity from plants performing much worse to those performing better than the parental lines of the cross. In the parental species, a nearly linear correlation was observed between photochemical chl fluorescence quenching (qP) and O2 evolution over a wide temperature range. Across the F3 generation, still a weak correlation between the two parameters was found at 20 °C, but not at 10 °C, when measured under identical conditions. This indicates that the fraction of open reaction centres could at least in part be adjusted to the photosynthetic capacity of the individual genotype. However, the correlation was so weak, that the previously suggested use of qP as a selection criterion for chilling tolerance of photosynthesis in breeding programs is regarded as doubtful, as long as photosynthesis rates are not measured in addition. Quantum efficiency of Photosystem II (ΦPSII) was strongly dependent on qP both at 20 and at 10 °C measuring temperature and depended on the quantum efficiency of open reaction centres (F′v/F′m) at 20, but not at 10 °C. F′v/F′m, in turn, correlated negatively with the processes of energy dissipation by the mechanisms of non-photochemical quenching (qN), i.e. its fast-relaxing component (qF) and photoinhibitory quenching (qI).
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    Differential Distribution of Gonadotropin-Releasing Hormone Variants in the Brain of Hydrochaeris hydrochaeris (Mammalia, Rodentia) (1999)
    Springer
    Cellular and molecular neurobiology 19 (1999), S. 635-651 
    Details
    ISSN: 1573-6830
    Keywords: gonadotropin-releasing hormone ; HPLC ; radioimmunoassay ; mammalian ; capybara
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1.In a previous paper we reported evidence for the presence of mGnRH- and sGnRH-like peptides in the preoptic–hypothalamic region of the capybara Hydrochaeris hydrochaeris (Montaner et al., 1998). In that study, the presence of a cGnRH-II like molecule in olfactory bulb extracts was suggested. 2.The capybara, the largest living rodent in the world, belongs to the order Hystricomorpha, which is considered to be one of the oldest groups of rodents. Some authors consider that this group is the ancestor of all remaining rodents. 3.In this study we have characterized GnRH molecular variants found in extracts from the olfactory bulbs and the mesencephalic region of capybara. These regions represent the two GnRH neuronal systems: the terminal nerve–septopreoptic and the midbrain systems. 4.An indirect method combining reverse-phase high-performance liquid chromatography (RP-HPLC) and radioimmunoassay (RIA) was used to characterize GnRH variants. The analysis of both extracts with two different RIA systems revealed three immunoreactive GnRH peaks, coeluting with mGnRH, cIIGnRH, and sGnRH synthetic standards. These results were additionally supported by serial dilution studies with specific antisera. 5.To our knowledge this the first report on the presence of three GnRH variants in the brain of an eutherian mammal. These results suggest that, similarly to other vertebrates, the expression of multiple GnRH variants may also be a common pattern in mammals.
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    No feeding on Phaeocystis sp. as solitary cells (post-bloom period) by the copepod Temora longicornis in the coastal waters of the English Channel (1999)
    Springer
    Hydrobiologia 414 (1999), S. 13-23 
    Details
    ISSN: 1573-5117
    Keywords: Phaeocystis sp. ; grazing ; copepods ; pigments ; HPLC ; English Channel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A Phaeocystis sp. (Prymnesiophyceae) bloom regularly occurs in April–May in the Eastern English Channel. In the literature, views are divided about the in situ appetence of copepods for this alga. In a study carried out in the coastal waters off the bay of Somme, at the end of the bloom, from 29 of April to 1 of May 1996, HPLC pigment analysis on both gut algal pigments and algal pigments from the water column shows that Temora longicornis adults did not feed on single cells of Phaeocystis sp. Alternatively, T. longicornis ingested diatoms and the gut content was correlated with the diatom biomass in the water. More, T. longicornis fed selectively on Dinophyceae and Cryptophyceae, which were scarcely present in the food environment. An inverse relationship was found between the concentration of Phaeocystis sp. in seawater and both gut content and abundance of young stages (CI–CIII copepodites) of T. longicornis. These results suggest an unfavourable impact of Phaeocystis sp. post-bloom on both feeding activity and distribution of T. longicornis.
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    Host specificity of Metriona elatior, a potential biological control agent of tropical soda apple, Solanum viarum, in the USA (1999)
    Springer
    BioControl 44 (1999), S. 423-438 
    Details
    ISSN: 1573-8248
    Keywords: classical biological control ; Solanaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The leaf beetle Metriona elatior from Brazil-Argentina was screened in the Florida (USA)State quarantine facility as a potential biological control agent of tropical soda apple, Solanum viarum, a recently arrived weed species. Multiple-choice host-specificity tests were conducted in small cages (60 cm × 60 cm × 60 cm) using 95 plant species in 29 families. Adults fed heavily on the main target weed (S. viarum), and on turkey berry,Solanum torvum (noxious weed of Asiatic origin); fed moderately on red soda apple, Solanum capsicoides (weed of South American origin), and eggplant, Solanum melongena (economic crop); and fed lightly on aquatic soda apple, Solanum tampicense (weed of Mexican-Caribbean-Central American origin), and onsilverleaf nightshade, Solanum elaeagnifolium(native weed widely distributed). M.elatior adults laid 84 to 97% of their egg masses onS. viarum, and 3 to 16% on S. melongena. Non-choice host-specificity tests were also conducted in quarantine in which M. elatior adults and neonate larvae were exposed to 17 and 19 plant species, respectively. Tests with the neonates indicate that this insect was able to complete its development on S. viarum, S. torvum, S. melongena, and S. capsicoides. Although some adult feeding and oviposition occurred on S.melongena in quarantine on potted plants in small cages, no feeding or oviposition by M. elatiorwas observed in field experiments conducted in Brazil. Surveys in unsprayed S. melongena fields in Argentina and Brazil indicated that M. elatioris not a pest of S. melongena in South America. The evidence obtained from the South-American field surveys, Brazil open-field experiments, and Florida quarantine host specificity tests indicate that M. elatior causes significant feeding damage toS. viarum, and does not represent a threat to S. melongena crops in the USA. Therefore an application for permission to releaseM. elatior against S. viarum in the USA was submitted in October 1998.
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    Automation of sample pretreatment for liquid chromatography (1998)
    Springer
    Microchimica acta 128 (1998), S. 19-29 
    Details
    ISSN: 1436-5073
    Keywords: automation ; sample preparation ; chromatography ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The purpose of this review is to discuss the strategic problems of automating sample preparation (SP) for high performance liquid chromatography (HPLC). There is a general feeling that SP is the bottleneck of many HPLC procedures. Despite numerous reports of successful automation of SP, there are still many laboratories using manual or semiautomated SP procedures. This calls for a reevaluation of the present situation.
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    Combining immunoassays with chromatographic and electrophoretic separation techniques — a review (1998)
    Springer
    Microchimica acta 129 (1998), S. 19-27 
    Details
    ISSN: 1436-5073
    Keywords: competitive ; non-competitive ; homogeneous ; heterogeneous ; pre-column immunoassay ; post-column immunoassay ; sandwich ; epitope ; on-line immunoassay ; off-line immunoassay ; laser-induced fluorescence ; microchip system ; HPLC ; CE ; digoxin digoxigenin ; solid phases affinity column ; urine ; plasma ; ELISA ; FAB fragments ; estrogen ; leukotriene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The combination of immunoassays with separation techniques such as chromatography and electrophoresis can provide both selectivity and sensitivity that is competitive with any method currently available for molecular analysis. Immunoassays can be carried out on-line and off-line, pre and post separation. The on-line post separation mode is the most promising for routine analysis because of the high throughput that can be achieved but also provides the greatest challenge with regard to compatibility of the interfaced systems. This paper reviews the various approaches that have been researched from a practical immunochemical point of view with emphasis on the special problems incurred with matrix compatibility for on-line post separation systems.
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    Analysis of fossil pigments from algae and bacteria in meromictic Lake Fidler, Tasmania, and its application to lake management (1998)
    Springer
    Journal of paleolimnology 19 (1998), S. 1-22 
    Details
    ISSN: 1573-0417
    Keywords: fossil pigments ; meromixis ; Lake Fidler ; Tasmania ; HPLC ; Mass Spectrometry ; lake management ; algae ; bacteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract Lake Fidler is an ectogenic meromictic lake with a monimolimnion maintained by periodic incursions of brackish water from the lower Gordon River estuary. A dam across the middle reaches of the Gordon River has restricted these incursions of brackish water and meromictic stability has rapidly declined. A palaeolimnological study was carried in order to assess the historical development of meromixis and the impact of the dam on the microbiological communities in the lake. Fossil pigments in a 17 m sediment core were analysed using reverse phase high performance liquid chromatography (rp-HPLC) and mass spectrometry (MS). In addition, taphonomic studies of pigment production, deposition and degradation in the water column and surface sediments were used to identify planktonic and benthic pigment degradation processes and constrain the stratigraphic interpretation. Results comparing the pigment composition of pelagic sediment traps and littoral surface sediments indicated that the core from the centre of the lake would permit a historical reconstruction of planktonic bacterial and algal communities. Marked increases in prokaryotic pigments ca 3500 yr B.P. suggested the possible colonisation of a chemocline by phototrophic bacteria. Further changes in chlorophyll: carotenoid ratios and changes in relative abundances of both chlorophyll a and bacteriochlorophyll c derivatives also indicated that a change in the depositional environment had occurred; possibly due to altered stratification or anoxia. From this we infer the onset of either intermittent or permanent meromixis. Further increases in prokaryotic pigment abundance suggested that the present state of permanent meromixis was firmly established by 2070 ±50 14C yr B.P., and diatom analysis confirmed the development of a stable mixolimnion. High resolution studies of the top 10 cm of sediments measured pigments in mean concentrations of 15.1 ng g-1 with a mean S.D. of only 2.78 indicating little change in pigment abundance since the construction of the dam. Thus, Lake Fidler still retains most of the features of meromixis. However, evidence from nearby Lake Morrison and Sulphide Pool has shown that any further declines in meromictic stability will cause a rapid reversion to holomixis. Palaeolimnological evidence from the early stages of meromictic development of Lake Fidler suggests that such reversion to holomixis may not permanently eliminate all the microbiological communities, and that, given time, they may return and prosper with re-establishment of a suitable chemocline. These studies will guide recommendations for a management strategy to prevent the further decay of meromixis in the Gordon River lakes.
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    URL: http://dx.doi.org/10.1023/A:1007909018527
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    Antibacterial activity of marine-derived fungi (1998)
    Springer
    Mycopathologia 143 (1998), S. 135-138 
    Details
    ISSN: 1573-0832
    Keywords: antibiotics ; HPLC ; marine penicillia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A total of 227 marine isolates of ubiqituous fungi were cultivated on different media and the secondary metabolite content of the extracts (ethyl acetate/chloroform/methanol 3 : 2 : 1) characterized by HPLC. The fungi were secured from animals, plants and sediments of Venezuelan waters (0–10 m) including mangroves and lagoonal areas. The extracts were tested for antibacterial activity. A total of 7 were active towards Vibrio parahaemolyticus and 55 towards Staphylococcus aureus, representing 18 different fungal species from 8 ascomycetous genera. For 61 strains of Penicillium citrinum antibacterial activity correlated well with content of secondary metabolites as measured by HPLC. Thirteen isolates of Penicillium steckii produced very similar profiles of secondary metabolites and 6 of these had activity against either V. parahaemolyticus or S. aureus or both.
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    URL: http://dx.doi.org/10.1023/A:1006961500325
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    Analysis of Fusarium toxins in maize and wheat using thin layer chromatography (1998)
    Springer
    Mycopathologia 142 (1998), S. 107-113 
    Details
    ISSN: 1573-0832
    Keywords: Deoxynivalenol ; Fumonisin B1 ; Zearalenone ; TLC ; HPLC ; ELISA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Thin layer chromatography (TLC) methods for identifying and quantifying deoxynivalenol (DON), fumonisin B1 (FB1) and zearalenone in grain samples were compared to immunoassay (ELISA) and high performance liquid chromatography (HPLC) methods to determine the reliability of the less expensive TLC. There was a very good agreement between levels of DON measured by TLC and competitive-direct ELISA, and between levels of fumonisin B1 measured by TLC and HPLC, over a wide range of concentrations. Correlation coefficients (Pearson's) were 0.978, 0.914 and 0.953 for DON in maize, DON in wheat and FB1 in maize respectively. A lower correlation coefficient (r = 0.672) was obtained when zearalenone was quantified by TLC and HPLC. Possible reasons for this are discussed. A cost comparison of the various methods revealed that TLC was the least expensive for sample analysis. It is recommended that researchers choose which analytical method to use based upon individual considerations of cost and precision.
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    URL: http://dx.doi.org/10.1023/A:1006937720711
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    Variability of some phenolic acids in phloem of 1-year-old shoots of Scots pine trees growing with Heterobasidion annosum (Fr.) Bref. (1998)
    Springer
    Trees 12 (1998), S. 230-235 
    Details
    ISSN: 0931-1890
    Keywords: Key words Scots pine ; Phenolic acids ; HPLC ; Heterobasidion annosum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  HPLC chromatographic analyses of some phenolic acids in phloem of 1-year-old shoots sampled from 32 trees of eight Polish provenances of Scots pine (Pinus sylvestris L.) growing under conditions of annosum root [Heterobasidion annosum (Fr.) Bref.] are discussed. Considerable quantitative and qualitative differentiation was found among individual trees. The variability of trees was estimated with regard to the level of phenolic acids and correlations were established in order to assess the character of their joint occurrence in shoot phloem. In view of pathogen presence, the content of phenolic acids varies between individuals depending upon the genotype of pine, the stage of development of the disease and upon the effect of tree growth conditions.
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    URL: http://dx.doi.org/10.1007/PL00009713
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    Photoinhibitory damage is modulated by the rate of photosynthesis and by the photosystem II light-harvesting chlorophyll antenna size (1998)
    Springer
    Planta 205 (1998), S. 288-296 
    Details
    ISSN: 1432-2048
    Keywords: Key words: Chlorophyll antenna size ; D1 protein ; Dunaliella (photoinhibition) ; Electron transport ; Photoinhibition ; Photosystem II
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. We investigated the effect of photosynthetic electron transport and of the photosystem II (PSII) chlorophyll (Chl) antenna size on the rate of PSII photoinhibitory damage. To modulate the rate of photosynthesis and the light-harvesting capacity in the unicellular chlorophyte Dunaliella salina Teod., we varied the amount of inorganic carbon in the culture medium. Cells were grown under high irradiance either with a limiting supply of inorganic carbon, provided by an initial concentration of 25 mM NaHCO3, or with supplemental CO2 bubbled in the form of 3% CO2 in air. The NaHCO3-grown cells displayed slow rates of photosynthesis and had a small PSII light-harvesting Chl antenna size (60 Chl molecules). The half-time of PSII photodamage was 40 min. When switched to supplemental CO2 conditions, the rate of photodamage was retarded to a t1/2 = 70 min. Conversely, CO2-supplemented cells displayed faster rates of photosynthesis and a larger PSII light-harvesting Chl antenna size (500 Chl molecules). They also showed a rate of photodamage with t1/2 = 40 min. When depleted of CO2, the rate of photodamage was accelerated (t1/2  = 20 min). These results indicate that the in-vivo susceptibility to photodamage is modulated by the rate of forward electron transport through PSII. Moreover, a large Chl antenna size enhances the rate of light absorption and photodamage and, therefore, counters the mitigating effect of forward electron transport. We propose that under steady-state photosynthesis, the rate of light absorption (determined by incident light intensity and PS Chl antenna size) and the rate of forward electron transport (determined by CO2 availability) modulate the oxidation/reduction state of the primary PSII acceptor QA, which in turn defines the low/high probability for photodamage in the PSII reaction center.
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    URL: http://dx.doi.org/10.1007/s004250050323
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    Production of a hepatoprotective cerebroside from suspension cultures of Lycium chinense (1998)
    Springer
    Plant cell reports 18 (1998), S. 252-254 
    Details
    ISSN: 1432-203X
    Keywords: Key wordsLycium chinense ; Hepatoprotective cerebroside ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Suspension cultures derived from Lycium chinense Miller seedlings produced significant amounts of a hepatoprotective cerebroside. Callus was induced from the stem of aseptic seedlings of L. chinense and maintained on MS solid media supplemented with 1.0 ppm 2,4-D and 0.1 ppm kinetin. Suspension cultures were established, and the cells were grown in the same liquid media in the dark. Lyophilized cells were extracted with a combined reagent of chloroform and methanol (2:1, v/v). An aqueous suspension of the evaporated cell extract was partitioned with chloroform, and the chloroform layer was subjected to silicic acid column chromatography followed by semi-preparative reverse phase C8 high pressure liquid chromatography. The purified compound showed hepatoprotective activity comparable to that shown by silymarin, and the structure was identified as 1-O-(β-d-glucopyranosyl)-(2S,3R,4E,8Z)-2-N-2′-hydroxy-(palmitoyl)-4,8-sphingadiene on the basis of spectral data. The content of the compound in cultured cell was tenfold higher than that of the fruit of L. chinense. The biosynthesis of the compound in cultured cell systems appears to parallel cell growth.
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    URL: http://dx.doi.org/10.1007/s002990050566
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    A novel thaumatin-like protein gene of tobacco is specifically expressed in the transmitting tissue of stigma and style (1998)
    Springer
    Sexual plant reproduction 11 (1998), S. 251-256 
    Details
    ISSN: 1432-2145
    Keywords: Key words Nicotiana tabacum ; Pathogenesis related protein ; Solanaceae ; Stigmatic exudate ; Thaumatin-like protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A cDNA clone that encodes a novel thaumatin-like glycosylated protein, SE39b, which constitutes one of the major proteins of the stigmatic exudate of tobacco, was isolated and sequenced. The deduced amino acid sequence of SE39b shows 37% identity with the pathogenesis-related protein of tobacco, PR-R1, and 52% identity with the thaumatin-like protein (TLP) of Arabidopsis. All 16 cysteine residues are conserved in SE39b as they are in all TLPs. Three potential glycosylation sites found in SE39b were consistent with a previous finding that concanavalin A has an affinity for SE39b. Northern blot and in situ analyses demonstrated that the gene was specifically expressed in the transmitting tissue of the stigma and style, and the transcript amounts reached maximum levels at anthesis. mRNA from orthologs of the gene of SE39b was detected in species of Cestreae, but not in species of Solaneae. SE39b should be categorized as a PR-like glycoprotein which is developmentally regulated and specifically expressed in the transmitting tissue of the stigma and style, such as Sp41 [(1,3)-β-glucanase], SK2 and Chi2;1 (chitinase).
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    URL: http://dx.doi.org/10.1007/s004970050150
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    Production of an Expression System for a Synaptobrevin Fragment to Monitor Cleavage by Botulinum Neurotoxin B (1998)
    Springer
    The protein journal 17 (1998), S. 453-462 
    Details
    ISSN: 1573-4943
    Keywords: Botulinum neurotoxin ; synaptobrevin ; thioredoxin ; recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Recombinant DNA techniques were used to develop an expression system for a 51-amino acid peptide fragment that encompasses residues 44–94 of human synaptobrevin 2. This protein is associated with secretory vesicles of nerve terminals and is a substrate for four of the seven serotypes of botulinum neurotoxin (BoNT). The DNA for the recombinant peptide was amplified by the polymerase chain reaction and cloned into the pTrxFus vector. The resulting synaptobrevin peptide was expressed as a thioredoxin fusion protein in E. coli and released into the medium by osmotic lysis. The 18.7-kDa thioredoxin-synaptobrevin protein, designated as TSB-51, is intended for use in a cell-free assay to test potential inhibitors of BoNT/B-mediated proteolysis of synaptobrevin with the ultimate aim of developing clinically effective therapeutic agents to counteract botulism. Incubation of TSB-51 with the purified light chain of BoNT/B resulted in proteolysis which was evident within 30 min and increased with time until completion (∼4 hr). Cleavage of TSB-51 appeared to be at the appropriate BoNT/B cleavage site as indicated by a reduced intensity of the 18.7-kDa band and the appearance of a band at 16.4 kDa on Tris-tricene polyacrylamide gradient gels. The concentration of free Zn2+ had a significant effect on the cleavage rate; low Zn2+ concentrations stimulated substrate cleavage, whereas high concentrations were inhibitory. Cleavage was not significantly depressed by the naturally occurring metalloprotease inhibitor phosphoramidon when tested at concentrations up to 5 mM. TSB-51 appears to be a useful substrate for studying BoNT/B and is expected to aid in the discovery of effective BoNT inhibitors.
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    URL: http://dx.doi.org/10.1023/A:1022570518330
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    Purification of combinatorial libraries (1998)
    Springer
    Molecular diversity 4 (1998), S. 47-52 
    Details
    ISSN: 1573-501X
    Keywords: chromatography ; HPLC ; library ; purification ; SPE
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract In the early days of combinatorial chemistry, much attention focused on preparation of large libraries for lead discovery. Recently, though, the focus has shifted toward smaller, more focused libraries for lead optimization. These focused libraries generally consist of individual discrete compounds. Biological assay requirements often require compounds of high purity, thus development of automated high throughput purification methods has received new attention in the past several years. This paper covers automated high throughput purification methods that have been applied to libraries of discrete compounds. Literature published through February 1998 is included. Purification methods discussed include extraction methods, scavenger methods, solid phase extraction, and preparative HPLC.
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    URL: http://dx.doi.org/10.1023/A:1009687711082
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    Thermoluminescence studies on the facultative crassulacean-acid- metabolism plant Mesembryanthemum crystallinum L. (1998)
    Springer
    Planta 205 (1998), S. 587-594 
    Details
    ISSN: 1432-2048
    Keywords: Key words: Crassulacean acid metabolism ; Mesembry-anthemum ; Photosystem II ; Thermoluminescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Thermoluminescence (TL) signals were measured from leaves of the facultative CAM (crassulacean acid metabolism) plant Mesembryanthemum crystallinum L.. Following the induction of CAM by salt treatment, a TL band at 46 °C was induced, which was charged by a single-turnover flash. The intensity of the 46 °C-band depends on the number of excitation flashes and oscillates with a period of four. A similar band was induced in C3 plants by far-red illumination. Under CAM conditions, the intensity of the 46 °C-band underlies a diurnal rhythm. The maximal intensity of the 46 °C-band is observed in the morning after onset of the light and in the evening. At around 12 a.m. it is suppressed. The intensity of the 46 °C-band relates to diurnal changes in the ratio of dihydroxy acetone phosphate/3-phosphoglycerate (DHAP/PGA) which is an indicator of the energy status of the chloroplast. During high-intensity illumination, the 46 °C-band disappears, but it is restored in the dark. We propose that the 46 °C-band is an indicator of the metabolic state of the leaf, originating from photosystem II centres initially in the S2(S3)QB oxidation state, in which the electron acceptor QB becomes reduced either by reverse electron flow or reduction of the plastoquinone pool via an NAD(P)H plastoquinone oxidoreductase. We present evidence that the redox state of the electron-transport chain is different under conditions of CAM compared to C3 metabolism and that changes induced by CAM can be monitored by measuring the amplitude of the 46 °C-band after flash excitation.
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    URL: http://dx.doi.org/10.1007/s004250050360
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    Mapping of the gene Nxphu that controls hypersensitive resistance to potato virus X in Solanum phureja IvP35 (1998)
    Springer
    Theoretical and applied genetics 96 (1998), S. 840-843 
    Details
    ISSN: 1432-2242
    Keywords: Key words Potato virus X ; Resistance gene ; Genetic mapping ; RFLP ; Solanaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The line IvP35 of the diploid (2n=2x=24) cultivated potato species Solanum phureja (family Solanaceae) expresses hypersensitive resistance (H) to potato X potexvirus (PVX). In this study, a diploid potato population was produced using IvP35 as the male parent and a diploid line of S. tuberosum (87HW13.7) as the female parent and tested for resistance to PVX. Data indicated that H to PVX in IvP35 is a dominant, monogenically inherited trait controlled by a single gene, named Nx phu , that is in a simplex condition (Nxnx). RFLP analysis carried out on the progeny lines revealed 4 markers (CT220, TG328, CT112 and TG424) from the long arm of chromosome IX that were linked to the hypersensitive phenotype; the closest linkage was observed with the marker TG424. Previous authors have shown that the same region of chromosome IX contains the gene Sw-5 for resistance to tomato spotted wilt tospovirus in Lycopersicon peruvianum (Solanaceae).
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    URL: http://dx.doi.org/10.1007/s001220050810
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    PsbY, a novel manganese-binding, low-molecular-mass protein associated with photosystem II (1998)
    Springer
    Molecular genetics and genomics 260 (1998), S. 56-68 
    Details
    ISSN: 1617-4623
    Keywords: Key words PsbY ; Photosystem II ; Gene duplication ; Polyprotein ; Topogenesis ; L-Arginine metabolizing enzyme
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe two related manganese-binding polypeptides with L-arginine metabolizing enzyme activity that can be detected as distinct components (designated PsbY-A1 and PsbY-A2, previously called L-AME) in membranes containing Photosystem II (PS II) from spinach. The polypeptides are bitopic and appear to exist in a heterodimeric form, but only in the chlorophyll a/b lineage of plants. Both proteins are encoded in the nucleus. In spinach and in Arabidopsis thaliana they are both derived from a single-copy gene (psbY) that is translated into a precursor polyprotein of approximately 20 kDa. The processing of the polyprotein is complex and includes at least four cleavage steps. Both polypeptides are exposed N-terminally to the lumenal and C-terminally to the stromal face of the thylakoid membrane.
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    URL: http://dx.doi.org/10.1007/s004380050870
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    Flow cytometric measurement of nuclear DNA content inCapsicum (Solanaceae) (1998)
    Springer
    Plant systematics and evolution 209 (1998), S. 85-91 
    Details
    ISSN: 1615-6110
    Keywords: Solanaceae ; Capsicum ; DNA content ; genome size ; flow cytometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nuclei were isolated from leaf tissue of differentCapsicum species and the relative fluorescence intensity was measured by flow cytometry after propidium iodide staining.Pisum sativum nuclei with known nuclear genome size (9.07 pg) were used as internal standard to determine nuclear DNA content of the samples in absolute units. The 2C DNA contents ranged between 7.65 pg inC. annuum and 9.72 pg inC. pubescens, and the general mean of the genus was 8.42 pg. These values correspond, respectively, to 1C genome size of 3.691 (C. annuum), 4.690 (C. pubescens) and 4.063 (general mean) Mbp. In general, white-flowered species proved to have less DNA, with the exception ofC. praetermissum, which displayed a 2C DNA content of 9.23 pg. It was possible to divide the studied species into three main groups according to their DNA content, and demonstrate differences in DNA content within two of the three species complexes established on the basis of morphological traits.
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    URL: http://dx.doi.org/10.1007/BF00991526
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    Pollen morphology and functional dioecy inSolanum (Solanaceae) (1998)
    Springer
    Plant systematics and evolution 210 (1998), S. 113-139 
    Details
    ISSN: 1615-6110
    Keywords: Solanaceae ; Solanum ; Evolution ; male sterility ; breeding system ; dioecy ; inaperturate pollen ; pollen development ; palynology ; plant reproduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dioecy has evolved independently several times in the large, mostly tropical genusSolanum. In all cases of dioecy inSolanum functionally male flowers have normal anthers, normal pollen and reduced stigmas while functionally female flowers have stigmas and anthers that appear normal but contain non-functional, usually inaperturate pollen. The inaperturate pollen has living cytoplasm, but apparently never germinates and it has been hypothesised that the pollen in these functionally female flowers is retained as a pollinator reward. Pollen morphology is compared in twelve of the thirteen known dioecious species ofSolanum, and some stages in the the development of inaperturate pollen in the anthers of functionally female flowers ofSolanum confertiseriatum of Western Ecuador are examined. Observations on the development and morphology of inaperturate pollen in functionally female flowers ofSolanum are related to hypotheses about the evolution of dioecy in the genus.
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    URL: http://dx.doi.org/10.1007/BF00984731
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    Free amino acid analysis of five microalgae (1998)
    Springer
    Journal of applied phycology 10 (1998), S. 131-134 
    Details
    ISSN: 1573-5176
    Keywords: Free amino acids ; HPLC ; microalgae ; Tetraselmis suecica ; Isochrysis galbana ; Thalassiosira sp. ; Skeletonema costatum ; Chaetoceros calcitrans ; cosmetology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The HPLC separation of fluorescent o-phtaldialdehyde (OPA) derivatives has been applied to the assay of free amino acids from five microalgae commonly used in aquaculture: Tetraselmis suecica, Skeletonema costatum, Chaetoceros calcitrans, Thalassiosira sp. and Isochrysis galbana, as part an assessment of their potential use in cosmetic products. Thirteen free amino acids were analyzed using High Performance Liquid Chromatography. There were considerable differences between species. However, four amino acids were responsible for more than 60% total concentration in all species: ASP, GLU, ARG and TYR; the next most important (accounting for less than 30%) were: ALA, VAL, PHE and LYS.
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    The photosystem II subunit CP29 can be phosphorylated in both C3 and C4 plants as suggested by sequence analysis (1998)
    Springer
    Plant molecular biology 36 (1998), S. 11-22 
    Details
    ISSN: 1573-5028
    Keywords: chlorophyll a/b protein ; CP29 ; Phosphorylation ; Photosystem II ; cold stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The CP29 subunit of Photosystem II is reversibly phosphorylated in Zea mays upon exposure to high light in the cold (Bergantino et al., J Biol Chem 270 (1995) 8474–8481). This phenomenon was previously proposed to be restricted to C4 plants. We present the complete sequence of the CP29 protein, deduced from a maize Lhcb4 cDNA clone, and its comparison with the previously known Lhcb4 sequences of two C3 plants: Hordeum vulgare and Arabidopsis thaliana. Despite the relatively low degree of homology in their amino-terminal region, i.e. the part of the molecule which is phosphorylated in maize, the three polypeptides conserve consensus sequences for the site of phosphorylation. We proved by immunoblotting and 33P-labelling that the same post-translational modification occurs in barley. Being thus common to C3 and C4 plant species, the phosphorylation of this minor antenna complex of Photosystem II appears now as a widespread phenomenon, possibly part of the phosphorylation cascade which signals the redox status of the plastoquinone to the nuclear transcription apparatus. Arabidopsis plants do not show phosphorylation of CP29 in the same conditions, but other low-molecular-weight phosphoproteins, whose role need to be elucidated, become evident.
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    URL: http://dx.doi.org/10.1023/A:1005904527408
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    The potential of AFLPs in biosystematics: A first application inSolanum taxonomy(Solanaceae) (1998)
    Springer
    Plant systematics and evolution 210 (1998), S. 87-103 
    Details
    ISSN: 1615-6110
    Keywords: Solanaceae ; Solanum ; AFLP markers ; DNA fingerprinting ; phylogeny ; polyploids ; potato ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using the AFLP technique highly informative DNA fingerprints were generated from 19 taxa ofSolanum sect.Petota (potatoes) and three taxa ofSolanum sect.Lycopersicum (tomatoes). Both phenetic and cladistic analyses were conducted from the individual genotypic level to the species level. An AFLP fingerprint, using a combination of suitable AFLP primers, generated 12 to 71 scorable fragments per genotype which was sufficient for taxonomic interpretation. The classifications based on the molecular markers were generally in agreement with current taxonomic opinions. Unexpectedly,S. microdontum was associated with ser.Megistacroloba rather than with ser.Tuberosa, andS. demissum (ser.Demissa) and species of ser.Acaulia appeared closely affiliated. AFLP is an efficient and reliable technique to generate biosystematic data and therefore a promising tool for evolutionary studies.
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    URL: http://dx.doi.org/10.1007/BF00984729
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    In vitro stem elongation of sweet pepper in media containing 24-epi-brassinolide (1998)
    Springer
    Plant cell, tissue and organ culture 53 (1998), S. 79-84 
    Details
    ISSN: 1573-5044
    Keywords: Capsicum annuum L. regeneration ; brassinolide ; Solanaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In vitro regeneration of sweet pepper (Capsicum annuum L. cvs Jupiter and Pimiento Perfection) has been performed via direct organogenesis. The resulting shoot-buds were placed on media containing 24-epi-brassinolide (EBR) 0.1 μM, a plant steroid lactone, in the presence or absence of zeatin 9.1 μM plus GA3 5.2 μM for further stem elongation. Different responses to these treatments were recorded depending upon the protocols used and the genotypes tested. It appears that EBR does not always act directly on stem elongation but may be an elicitor and/or an enhancer of elongation in concert with endogenous and other exogenously added growth regulators. Elongated shoots were easily rooted with alpha-naphtalenacetic acid 0.5 μM (0.1 mgl-1) and transfered to soil, and following acclimation were taken to maturity in the greenhouse.
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    URL: http://dx.doi.org/10.1023/A:1006071803855
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    Indifference of potato anther culture to colchicine and genetic similarity among anther-derived monoploid regenerants determined by RAPD analysis (1998)
    Springer
    Plant cell, tissue and organ culture 53 (1998), S. 49-58 
    Details
    ISSN: 1573-5044
    Keywords: androgenesis ; secondary embryogenesis ; Solanaceae ; Solanum phureja
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Effects of colchicine on androgenesis of diploid potato (Solanum phureja Juz. & Buk.) and ploidy of anther-derived plants were examined in three experiments. In the first, no significant difference was found for mean embryos per anther of an interspecific potato clone after application of five colchicine treatments (0, 25, 50, 100 and 200 mg l-1) for 24 h to freshly excised anthers containing late uninucleate microspores. The same colchicine treatments were applied to six hybrid potato families in the second experiment. Families differed for number of embryos per anther and embryo regeneration frequency; however, androgenic response did not differ significantly among colchicine treatments. The 312 regenerated plants included 233 (75%) monoploids. The third experiment examined durations (0, 90 s vacuum infiltration, 24, 48 and 72 h) of high colchicine treatment (200 mg l-1) on anther culture of seedlings representing one family. Mean embryos per anther, though not statistically significant, ranged from 0.96 to 1.90 for 48 h colchicine and 90 s vacuum infiltration, respectively. There were 126 plants regenerated of which 62% were monoploid. Frequency of monoploid plants regenerated from colchicine treatments did not differ significantly. RAPD analysis was conducted on 26 anther-derived monoploids of one family, based on common flasks of origin. The 13 decamer primers revealed 54 polymorphic loci. These were used to characterize the monoploids genetically. From one flask, two pairs of monoploids among six examined were genetically indistinguishable. Examination of a second and third flask revealed, six of seven and three of seven monoploids that were genetically indistinguishable. These data suggest the regeneration of genetic clones within flasks and may indicate the occurrence of secondary embryogenesis during anther culture.
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    URL: http://dx.doi.org/10.1023/A:1006099423651
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