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  • 1
    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.
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  • 2
    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.
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  • 3
    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.
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  • 4
    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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  • 5
    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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  • 6
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    Biologia plantarum 43 (2000), S. 79-84 
    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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  • 7
    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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  • 8
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    Plant cell, tissue and organ culture 58 (1999), S. 133-140 
    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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  • 9
    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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  • 10
    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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  • 11
    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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  • 12
    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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  • 13
    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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  • 14
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    Mycopathologia 142 (1998), S. 107-113 
    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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  • 15
    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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  • 16
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    Plant cell reports 18 (1998), S. 252-254 
    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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  • 17
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    Journal of applied phycology 10 (1998), S. 131-134 
    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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  • 18
    ISSN: 1573-5079
    Keywords: carotenoid ; chlorophyll b formation ; chlorophyllide a esterification ; accumulation of photosynthetic pigments ; HPLC ; protochlorophyllide a
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chlorophyll and carotenoid variations of 2-d-old and 10-d-old bean leaves (Phaseolus vulgaris var Red Kidney) were analyzed by HPLC during the first photoperiod of greening (16 h light + 8 h dark). The HPLC method used is suitable for the separation of cis- and trans-carotenoid isomers, Pchlide a and Chlide a as well as their esters. The main results are (1) before illumination the composition of the carotenoid pool is similar at the two developmental stages; (2) non-illuminated 2-d-old leaves are devoid of Pchlide a ester; (3) chlorophyll and carotenoid accumulation in 2-d-old leaves presented a lag phase twice longer than observed in 10-d-old ones; (4) Chlide a seems directly esterified to Chl a in 2-d-old leaves whereas esterification requires four steps in 10-d-old leaves and, (5) the kinetics of Chl and carotenoid accumulation are different at the two investigated developmental stages.
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  • 19
    ISSN: 0884-3996
    Keywords: bioluminescence ; adrenalin ; noradrenalin ; photophores ; HPLC ; mesopelagic fish ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The presence of adrenalin (E) and noradrenalin (NE) was found by HPLC both in the photophores and at other tissue levels of numerous species of mesopelagic fish in The Strait of Messina, with the aim of determining the incidence of these catecholamines in photophores, in light transmission and the eventual presence at other tissue levels. © 1998 John Wiley & Sons, Ltd.
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  • 20
    ISSN: 0730-2312
    Keywords: HMG-CoA ; MVA ; HPLC ; dolichol-like lipids ; DNA synthesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Substantial evidence has suggested that a nonsterol product of mevalonic acid (MVA) is essential for the initiation of DNA synthesis in mammalian cells. Several possible isoprenoid candidates have been suggested, but the identity of this compound still remains unknown. In this study we have isolated and purified MVA products from SV40-transformed human fibroblasts and identified fractions with a growth-stimulatory effect. The cells were labelled with [14C]MVA in the presence of inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. After lipid extraction, the [14C]MVA-labelled lipids were subjected to high performance liquid chromatography and size-exclusion chromatography, and the effect of the fractionated eluate on the DNA synthesis of arrested MVA-depleted target cells was tested. Thereby we found a fraction of [14C]MVA-labelled lipids with a substantial stimulatory effect on DNA synthesis. The chromatographic behavior suggested that the growth-stimulating fractions contained dolichol-20. This was confirmed by mass spectrometric analysis. Similar results were obtained when lipids from hepatocellular carcinoma cells and a sample from breast tumor were isolated and analyzed by the same procedure. The mechanisms by which these compounds induce DNA synthesis are unknown. Recent data obtained in our laboratory have provided evidence that dolichyl groups are covalently linked to tumor cell proteins, which implicates a new biological function for long-chain polyisoprenoid alcohols (Hjertman et al. [1997] FEBS Lett 416:235-238). In this study we demonstrate that tumor cells containing dolichol-like growth-stimulatory lipids also contained dolichylated proteins. This raises the question whether the growth-stimulatory dolichol-like lipids serve as substrates for the dolichylation reaction. J. Cell. Biochem. 71:502-514, 1998. © 1998 Wiley-Liss, Inc.
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  • 21
    ISSN: 1573-0417
    Keywords: palaeolimnology ; pigments ; massspectrometry ; HPLC ; carotenoids ; chlorophylls ; bacteriochlorophylls ; biomarkers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract Accurate identification of fossil pigments is essential if they are to be used as biomarker compounds in palaeolimnological studies. In recent years High Performance Liquid Chromatography (HPLC) has greatly enhanced the efficiency with which fossil pigments can be characterised and quantified. Using HPLC, undegraded pigments are typically identified through retention times, absorbance spectra and co-chromatography with authentic reference standards. However, lake sediments may also contain degraded pigments for which there are often no standards, and which may be difficult to identify using HPLC alone. In this study, we submitted HPLC fractions of fossil pigments and pigment derivatives collected from a meromictic lake in south west Tasmania, to a combination of Mass Spectrometry (MS) techniques including Electron Impact (EI) and static Liquid Secondary Ion MS (LSIMS) to identify their molecular ion characteristics and organic chemical composition. Mass Spectrometry permitted the detection of specific mass ions which were used to verify the identity of pigments and their derivatives. These included five carotenoids, chlorophyll a and derivatives, three previously described bacteriochlorophyll c derivatives with molecular weights of 770, 784, and 802, and two undescribed derivatives of bacteriochlorophyll c with molecular weights of 766 and 788. With these improved identifications we speculate on the pathways and modes of pigment degradation in the lake and asses the value of the degraded pigments as biomarkers. The use of MS permitted the identification of a greater number of signature pigments of algal and bacterial communities thus increasing the palaeolimnological value of the sediments. These methods are best applied in fossil pigment studies where there are a large number of unknown pigments and pigment degradation products, and where there are no authentic standards for co-chromatography. Practical suggestions for pigment MS are included in the discussion.
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  • 22
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    Cell & tissue research 288 (1997), S. 127-134 
    ISSN: 1432-0878
    Keywords: Key words: Taurine ; Immunohistochemistry ; HPLC ; Amino acid neurotransmitters ; Renilla koellikeri (Cnidaria)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. A quantitative evaluation of putative amino acid neurotransmitters in sea pansy polyps by high-performance liquid chromatography indicates that the taurine content exceeds that of other amino acids by a 100-fold. The cellular source of this taurine was investigated by immunohistochemistry with two polyclonal antisera raised in rabbit, one against a glutaraldehyde-polylysine-taurine conjugate and the other against a succinylated ovalbumin-carbodiimide-taurine conjugate. Taurine-immunoreactive neurons were localized in a perioral subectodermal nerve net and in the zooid nerve net of the endodermal retractor muscle of the polyp mesenteries. Double labeling experiments revealed that taurine immunostaining does not colocalize with Phe–Mat–Ang–Phe –NH2 FMRFamide immunoreactivity. In addition, strong taurine immunoreactivity was found in nematocytes and other ectodermal cells, in myoepithelial cell bodies of the endoderm, and in calcareous spicule-producing cells of the colonial tissue mass. The limited distribution of neuronal taurine immunostaining to nerve nets associated with muscle systems subtending autozooid polyp retraction supports a role for taurine as a neuromuscular transmitter for this protective reflex. In contrast, the widespread distribution of taurine immunoreactivity in nematocytes and in other nonneuronal cells points to additional cellular functions of taurine, one of which may be to mediate responses to osmotic or metabolic stress.
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  • 23
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    Plant systematics and evolution 208 (1997), S. 1-9 
    ISSN: 1615-6110
    Keywords: Fabaceae ; Vicia faba ; V. kalakhensis ; Seed albumins ; HPLC ; taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previously reported electrophoretic seed albumin data have shown an unexpected association ofVicia faba withV. kalakhensis. In the present work, seed albumins ofV. faba (subsp.paucijuga and subsp.faba) were compared with those ofV. kalakhensis using ionexchange (IE) and reversed-phase (RP) high-performance liquid chromatography (HPLC). Two subspecies ofV. faba displayed similar seed albumin profiles. On the other hand, seed albumin profiles ofV. faba andV. kalakhensis showed no major protein peak in common either in IE-HPLC or RP-HPLC chromatograms. The reported differences in seed albumin composition ofV. faba andV. kalakhensis are consistent with other taxonomical data showingV. faba to be genetically distant from the wild relatives.
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  • 24
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    Hydrobiologia 352 (1997), S. 251-262 
    ISSN: 1573-5117
    Keywords: Red Sea ; PAHs ; oil pollution ; HPLC ; GC/MS ; fish ; origin ; toxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A detailed analytical study using combined normal phase high pressure liquid chromatography (HPLC), gas chromatography (GC) and gas chromatography/mass spectrometry (GC/MS) of Polynuclear Aromatic Hydrocarbons (PAHs) in fish from the Red Sea was undertaken. This investigation involves a preliminary assessment of the sixteen parent compounds issued by the U.S. Environmental Protection Agency(EPA). The study revealed measurable levels of Σ PAHs (the sum of three to five or six ring parent compounds) (49.2 ng g−1 dry weight) and total PAHs (all PAH detected) (422.1 ng g−1 dry weight) in edible muscle of fishes collected from the Red Sea. These concentrations are within the range of values reported for other comparable regions of the world. Mean concentrations for individual parent PAH in fish muscles were; naphthalene 19.5, biphenyl 4.6, acenaphthylene 1.0, acenaphthene 1.2, fluorene 5.5, phenanthrene 14.0, anthracene 0.8, fluoranthene 1.5, pyrene 1.8, benz(a)anthracene 0.4, chrysene 1.9, benzo(b)fluoranthene 0.5, benzo(k)fluoranthene 0.5, benzo(e)pyrene 0.9, benzo(a)pyrene 0.5, perylene 0.2, and indeno(1,2,3-cd)pyrene 0.1 ng g−1 dry weight respectively. The Red Sea fish extracts exhibit the low molecular weight aromatics as well as the discernible alkyl-substituted species of naphthalene, fluorene, phenanthrene and dibenzothiophene. Thus, it was suggested that the most probable source of PAHs is oil contamination originating from spillages and/or heavy ship traffic. It was concluded that the presence of PAHs in the fish muscles is not responsible for the reported fish kill phenomenon. However, the high concentrations of carcinogenic chrysene encountered in these fishes should be considered seriously as it is hazardous to human health. Based on fish consumption by Yemeni‘s population it was calculated that the daily intake of total carcinogens were 0.15 µg/person/day.
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  • 25
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    Invertebrate neuroscience 2 (1997), S. 253-260 
    ISSN: 1439-1104
    Keywords: Aplysia ; serotonin ; aging ; weight ; development ; HPLC
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    Topics: Biology
    Notes: Abstract The neurotransmitter serotonin (5-HT) plays an important role in a number of behaviors inAplysia californica some of which have been shown to vary with age. We were thus interested in examining the age-dependence of 5-HT inA. californica. Because animals of the same age can have very different weights, and weight alone is reliably known for wild-caught animals, we also examined the variation of 5-HT with weight. Serotonin was measured in the ring and abdominal ganglia combined, in lab-reared animals from 3 to 12 months post-hatch across a wide weight range. Serotonin increased rapidly from 4 to 6 months, and more slowly from 6 to 13 months. Serotonin scaled by soluble ganglion protein increased from 3 to 6–7 months, reached a maximum, and then decreased again. Serotonin, but not scaled 5-HT, increased significantly with weight across the whole weight range. Animals of the same weight, but different ages, had different 5-HT levels, as did young animals of the same age but different weight. Serotonin varied significantly with both age and weight, with the age-dependence being the more significant.
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  • 26
    ISSN: 1573-0832
    Keywords: Fumonisins ; o-phthalaldehyde ; HPLC ; postcolumn derivatization ; LC/MS
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    Topics: Biology , Medicine
    Notes: Abstract Fumonisins B1(FB1) and B2(FB2) were isocratically separated on a fluorocarbon column without using an ion pair reagent and nonvolatile buffer during the HPLC and were detected by an o-phthalaldehyde postcolumn derivatization system using a fluorescence detector. The minimum detectable concentrations of FB1 and FB2 in corn by this system were 0.01 μg/g and 0.01 μg/g, respectively. The separated fumonisins were further identified by a directly interfaced ion trap MS using electrospray ionization. FB1 and FB2 in naturally contaminated corn were identified in the selective ion monitoring mode at concentrations of 3.75μg/g and 1.44 μg/g, respectively.
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  • 27
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    Plant cell, tissue and organ culture 51 (1997), S. 83-87 
    ISSN: 1573-5044
    Keywords: Agrobacterium rhizogenes ; hairy root ; henna ; HPLC ; Lawsonia inermis ; lawsone ; micropropagation
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    Topics: Biology
    Notes: Abstract The improvement of axillary shoot formation of Lawsonia inermis L. cultured in vitro depended on the iron concentration in the culture medium. Regenerated shoots were rooted on a hormone-free half-strength Murashige and Skoog medium (1/2 MS) before transfer to greenhouse conditions. Determination of lawsone in the plant material was investigated using a new HPLC method. The results showed that lawsone accumulation in vivo is restricted to the aerial part of the plant. In addition, the possibility of inducing lawsone biosynthesis in root cultures was studied. Hairy root cultures were established by a co-culture method using leaf segments of L. inermis and Agrobacterium rhizogenes NCIB 8196. Of several basal media tested, the production of lawsone (0.13% dry weight) was only observed in hairy roots tissues incubated in the dark and cultured in 1/2 MS or MS media.
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  • 28
    ISSN: 1573-0778
    Keywords: adaptive control ; ammonia ; antithrombin III ; BHK cells ; fed-batch culture ; fuzzy control ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract An adaptive fuzzy controller was developed to control the glucose and glutamine concentrations in the reactor constant at the desired level. The parameter values of the controller change during the cultivation according to the culture phase which was detected by the lactate concentration. Cultivations with different glucose and glutamine set point concentrations of a recombinant BHK anchorage-dependent cell line were performed in a fed-batch reactor on-line connected with an HPLC system. Glucose and glutamine concentrations were satisfactorily controlled at each set point during all cultivation periods. Ammonia had a determining effect on productivity since it inhibited cell growth and protein specific production. Ammonia production increased with an increase of glutamine or a decrease of glucose set point concentrations, indicating the importance of glucose to glutamine ratio for the optimization of productivity in mammalian cell cultures.
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  • 29
    ISSN: 1570-7458
    Keywords: antagonist ; bioassay ; ecdysteroid ; HPLC ; steroid hormone receptor ; withanolide
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    Topics: Biology
    Notes: Abstract Sixteen withanolides isolated from Iochroma gesnerioides (Kunth) Miers (Solanaceae) have been assessed for their activities as ecdysteroid agonists and antagonists. None of the compounds showed any agonistic activity, but several showed significant antagonistic activity. With a 20-hydroxyecdysone concentration of 5 × 10−8 M, the ED50 values for 2,3-dihydro-3ξ-methoxywithaferin A, 2,3-dihydro-3ξ-methoxywithacnistine, 2,3-dihydro-3ξ-methoxyiochromolide and 2,3-dihydro-3ξ-hydroxywithacnistine are 3.5 × 10-5 M, 1 × 10−5 M, 5 × 10−6 M and 2.5 × 10−6 M, respectively.
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    Aquatic sciences 58 (1996), S. 241-252 
    ISSN: 1420-9055
    Keywords: HPLC ; chlorophylls ; phaeopigments ; carotenoids ; xanthophylls
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    Topics: Biology
    Notes: Abstract To determine some of the environmental effects that influence the relative proportions of pigments in algae, high pressure liquid chromatographic (HPLC) analysis was employed to determine the relative amounts of common photosynthetic pigments in batch cultures of the diatoms,Fragilaria crotonensis andThalassiosira pseudonana, and the green algae,Scenedesmus abundans, andHaematococcus pluvialis, illuminated for 12 hours each day. Similar analyses were conducted in five-day experiments during which cultures ofF. crotonensis andS. quadricauda were kept in continuous darkness. Comparing the results to those for controls continuing to receive the daily illumination indicated that the diatoms and green algae react similarly to light deficiency. The relative amounts of the main accessory pigment in the diatoms, fucoxanthin, and that in the green alga, apparently lutein, decreased as a reaction to a lack of illumination, while the total chlorophyll level in algae of both groups remained nearly constant. Quantitative differences induced by the experimental conditions were considerably less that those observed among different species of diatom or among the different green algae, however. Finally, cultures ofS. quadricauda were analyzed and then kept for 43 days without the addition of any nutrients. A proportion of the culture was kept for this period in perpetual darkness while another continued to receive 12 hours of illumination. The results show that considerable changes occur as the cultures age, and that these changes occur more slowly in the darkness. Some consequences of these findings for phytoplankton production studies based on analyses of photosynthetic pigments are discussed.
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  • 31
    ISSN: 1432-2242
    Keywords: RAPD-PCR ; HPLC ; Watermelon ; Citrullus lanantus (Thunb.) Mansf. ; Phenogram
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    Topics: Biology
    Notes: Abstract RAPD (random amplified polymorphic DNA) markers generated by 15 arbitrary decamers were used to determine the frequency of DNA polymorphism in 39 watermelon [Citrullus lanantus (Thunb.) Mansf.] germplasms. Of the 15 primers tested, all except 1 (primer 275) directed the amplification of polymorphic products. A total of 162 amplification products were generated across all 39 genotypes. Among the 162 fragments, 35 (21%) appeared to be reliable polymorphic markers. The mean value by marker difference in this comparison was 0.24, and the highest, 0.69. Eight RAPD markers could be utilized in the unique variety discrimination 8 watermelon genotypes. From the phenograms constructed by UPGMA based on the comparison of RAPD markers, four clusters were resolved. Each group was also characterized and identified with morphological and genetic characteristics for each genotype. The free sugars of the edible parts of watermelons were analyzed by HPLC (high-performance liquid chromatography). Results from the phylogenetic analysis of band sharing data were consistent with sweetness as measured by HPLC. In conclusion, RAPD assays can be used for providing alternative markers for identifying genotypes and quantitative characteristics in watermelon.
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  • 32
    ISSN: 1432-2242
    Keywords: Key words RAPD-PCR ; HPLC ; Watermelon ; Citrullus lanantus (Thunb.) Mansf. ; Phenogram
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  RAPD (random amplified polymorphic DNA) markers generated by 15 arbitrary decamers were used to determine the frequency of DNA polymorphism in 39 watermelon [Citrullus lanantus (Thunb.) Mansf.] germplasms. Of the 15 primers tested, all except 1 (primer 275) directed the amplification of polymorphic products. A total of 162 amplification products were generated across all 39 genotypes. Among the 162 fragments, 35 (21%) appeared to be reliable polymorphic markers. The mean value by marker difference in this comparison was 0.24, and the highest, 0.69. Eight RAPD markers could be utilized in the unique variety discrimination 8 watermelon genotypes. From the phenograms constructed by UPGMA based on the comparison of RAPD markers, four clusters were resolved. Each group was also characterized and identified with morphological and genetic characteristics for each genotype. The free sugars of the edible parts of watermelons were analyzed by HPLC (high-performance liquid chromatography). Results from the phylogenetic analysis of band sharing data were consistent with sweetness as measured by HPLC. In conclusion, RAPD assays can be used for providing alternative markers for identifying genotypes and quantitative characteristics in watermelon.
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  • 33
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    Journal of plant research 109 (1996), S. 223-230 
    ISSN: 1618-0860
    Keywords: Bacteriochlorophyllg′ ; Chlorophylla′ ; Green sulfur bacteria ; Heliobacteria ; HPLC ; PS1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Based on precise pigment analyses of a series of photosystem (PS) 1-type plants, a novel hypothesis is proposed that chiorophyll (Chl)a′ and bacteriochlorophyll (BChl)g′, the 132-epimers of Chla and BChlg, constitute the primary electron donors of PS1 and heliobacteria, respectively. Interestingly PS 1-type reaction centers do not have (B) Pheo but have Chl a-like pigments as primary electron acceptors.
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    Journal of applied phycology 8 (1996), S. 509-515 
    ISSN: 1573-5176
    Keywords: ADAM ; Dinophysis fortii ; dinophysistoxin-1 ; DSP ; HPLC ; okadaic acid ; scallop
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell densities of toxic phytoplankton species responsible for diarrhetic shellfish poisoning (DSP) were monitored at a sampling site in Mutsu Bay, Japan, in 1995.Dinophysis fortii almost completely dominated the toxic phytoplankton community. Okadaic acid (OA) and dinophysistoxin-1 (DTX1) contents in bothD. fortii cells and midgut glands of scallops collected at the same sampling site were determined by HPLC — fluorometry. DTX1 was detected fromD. fortii and scallops. The contents of DTX1 inD. fortii changed markedly during the experimental periods (5–252 pg cell−1). The highest concentration of DTX1 in the midgut glands of scallops coincided with the period of relatively high cell densities ofD. fortii with the highest content of DTX1 (252 pg cell−1). The results demonstrate that toxin content in the cells is an important factor affecting the toxicity of shellfish.
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  • 35
    ISSN: 1573-6830
    Keywords: catecholamines ; hypothalamus ; development ; uptake ; α-methyl-m-tyrosine ; 6-hydroxydopamine ; HPLC ; rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. The present study aimed to develop a pharmacological model of catecholamine (CA) depletion in the hypothalamus during the period of its morphofunctional development, i.e. in fetal and neonatal rats of both sexes. 2. In the first series of experiments, pregnant females and, hence, fetuses were systemically treated daily from the embryonic day (E) 13 to E20 with the inhibitor of the CA synthesis α-methyl-m-tyrosine. The CA concentrations were subsequently measured in the fetal hypothalamus at E21 by high performance liquid chromatography with electrochemical detection (HPLC-ED). In the second series of experiments, neonatal rats were injected with neurotoxin, 6-hydroxydopamine and/or α-methyl-m-tyrosine daily from the 2nd postnatal day (P2) to P10. 3. The HPLC-ED assay of hypothalamic catecholamines (CA's) at E21 and P11 showed that both in fetuses and neonates, α-methyl-m-tyrosine caused more than 50% depletion of hypothalamic noradrenaline and adrenaline, while the dopamine (DA) level remained unchanged. The combined treatment of neonatal rats with α-methyl-m-tyrosine and 6-hydroxydopamine resulted additionally in a 25% decreased level of DA. 4. The influence of CA deficiency on the developing hypothalamic CA system was further evaluated by measuring [3H]DA uptake by nervous tissuein vitro. 5. The CA deficiency caused a 50% drop of [3H]DA uptake by the hypothalamic tissue in treated fetuses suggesting a stimulating effect of CA's on the early development of the CA system. In pharmacologically treated neonatal rats [3H]DA uptake remained at the control level showing no influence of the CA deficiency on the developing CA system after birth. 6. The usefulness of the proposed pharmacological model for studying of CA influence on differentiating hypothalamic target neurons is discussed.
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    World journal of microbiology and biotechnology 12 (1996), S. 239-242 
    ISSN: 1573-0972
    Keywords: p-coumaric acid ; ferulic acid ; HPLC ; phenolic acid esterase assay
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A rapid, simple and sensitive method for detection of ferulic and p-coumaric acids using HPLC has been developed which can be used to determine the respective phenolic acid esterase activities of microorganisms. Prior concentration, purification or derivatization of the samples are not required. As little as 0.5 mg ferulic or p-coumaric acid/I could be detected and estimated in 〈 1 h. The method is specific for the two phenolic acids sice no interference by other components was observed.
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  • 37
    ISSN: 1573-1561
    Keywords: Papilio polyxenes ; Papilionidae ; Daucus carota ; Apiaceae ; oviposition ; leaf surface ; contact chemoreception ; HPLC ; flavonoid glycosides ; chlorogenic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Ovipositing black swallowtail butterflies,Papilio polyxenes, make their final host-selection decisions on the basis of compounds present on the leaf surface. Little information is available, however, on the chemistry of leaf surfaces. The purpose of this study was to develop a technique to extract and quantify the concentrations of compounds from the leaf surfaces ofDaucus carota, one of the main host species forP. polyxenes, with particular reference to compounds already identified as contact oviposition stimulants, namelytrans-chlorogenic acid (CA) and luteolin-7-O-(6″-O-malonyl)-β-d-glucopyranoside (L7MG), as well as its degradation product luteolin-7-glucoside (L7G). Plant surfaces were extracted by dipping leaves sequentially in pairs of solvents: (1) CHCl3-MeOH, (2) near-boiling H2O, (3) CHCl3-near-boiling H2O, and (4) CH2Cl2-CH2Cl2. The resulting extracts were fractionated and analyzed using high-performance liquid chromatography. The leaf-surface concentrations of each compound were calculated using regressions relating leaf surface area to leaf weight that were obtained from measurements of field-collected carrot plants. All four methods removed the three compounds from carrot leaf surfaces, but the solvent systems differed in effectiveness. The chloroform-near-boiling water solvent system performed better than the other solvent combinations, but not significantly so. This system also extracted the highest number of polar, UV-absorbing compounds. Methylene chloride was significantly less efficient than the other methods. An additional test confirmed that the chloroform-near-boiling water method removed compounds from the surface alone and probably not from the apoplast or symplast. Surface concentrations of CA (up to 600 ng/cm2 leaf surface) were substantially greater than those of the two flavonoid compounds. No clear seasonal trend in concentrations was evident from the limited number of sampling dates.
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    World journal of microbiology and biotechnology 12 (1996), S. 32-37 
    ISSN: 1573-0972
    Keywords: Bacillus thuringiensis subsp. darmstadiensis ; HPLC ; modified airlift reactor ; net-draft-tube ; thuringiensin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A net-draft-tube, modified airlift reactor and a stirred-tank reactor were used for thuringiensin production by Bacillus thuringiensis subsp. darmstadiensis growing with various concentrations of molasses. The optimum concentration of molasses for thuringiensin production in both reactors was 15 g/l. There was a 6 h delay in sporulation in the modified airlift reactor compared with that in the stirred-tank reactor. Thuringiensin yield in the modified airlift reactor (2.2 g/l) was consequently higher than that in the stirred-tank reactor (1.1 g/l).
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    Journal of comparative physiology 176 (1995), S. 761-771 
    ISSN: 1432-1351
    Keywords: Biogenic amines ; HPLC ; Trichoplusia ni ; Circadian rhythm
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Quantitative levels of melatonin and 5-hydroxytryptamine were measured over the scotophase in the protocerebrum, subesophageal ganglion, optic lobes, thoracic ganglia, and hemolymph of adult male cabbage looper moths, Trichoplusia ni, using HPLC with electrochemical detection. Melatonin levels were very low (s 〈 1 pmol) or undetectable during the photophase, but increased in all tissues during the dark. Lowest mean levels in the dark were observed in the optic lobes (0.3 to 0.7 pmols). Maximal mean levels in the protocerebrum (5.2 pmols) occurred in the early part of the scotophase, but in all other tissues (2.8 in the subesophageal ganglion; 9.5 in thoracic ganglia) and hemolymph (18 pg/μl) maximal mean levels were observed later in the dark. Levels of 5-hydroxytryptamine in each tissue, in contrast, were higher than melatonin levels in the photophase, and in the protocerebrum and thoracic ganglia decreased during the dark, but in the optic lobes and subesophageal ganglion remained unchanged. Further, decreases in 5-hydroxytryptamine during the dark were significantly lower than the increased levels of melatonin, suggesting that active synthesis of 5-hydroxytryptamine was occurring. In a second experiment, when measured from individuals in three different photoperiods (6∶18, 12∶12, 18∶6 light∶dark) maximum mean melatonin levels in the brain (protocerebral and subesophageal ganglia) peaked within the first 1.5 h of the dark and remained at measurable levels for the duration of the dark. In a third experiment, levels of melatonin in the brain and thoracic ganglia displayed rhythmicity in continuous dark conditions but not in continuous light, when compared with profiles obtained in a normal light ∶ dark regime.
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  • 40
    ISSN: 1573-4978
    Keywords: aminoacyl-tRNA synthetase ; embryogenesis ; HPLC ; total RNA ; transfer RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Total RNA as well as transfer RNA were quantified from mature ova apart from four different embryonic stages namely mid-cleavage, early gastrula, mid-gastrula and organogenesis of the freshwater teleostHeteropneustes fossilis. Total RNA as well as transfer RNA quantity follow a similar variation pattern, being maximum during mid-gastrulation. When analysed by total amino acid acceptance capacity, transfer RNA shows its maximum activity during mid-gastrulation. This coincides with the higher ratio of tRNA to total RNA at this stage. The relative aminoacylation capacity for Ser, Gly, Asn and Thr are found to be higher (9–34%) compared to that for other amino acids. Total tRNA, resolved into three peaks upon HPLC fractionation, shows a high cumulative peak area during mid-gastrulation and organogenesis. These results indicate a switch over of maternal to embryonic translation machinery during gastrulation.
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    Journal of applied phycology 7 (1995), S. 487-494 
    ISSN: 1573-5176
    Keywords: HPLC ; method ; algal pigments ; chlorophyll ; carotenoid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of columns (Nucleosil C18ODS, MZ-PAH, YMC-PACK C30), column properties (inner diameters of 4 mm, 3 mm and 2 mm, pore-width 10 nm and 30 nm) and eluents (methanol, acetonitrile, acetone, water) were tested on the separation of algal pigments. The length of columns was 250 mm and particle size was 5 μm. Flow rates and gradients were adjusted to optimize peak separation; remaining chromatographic conditions were kept constant. The resolution of chromatographic systems was tested with pigment standards and various algal cultures. Total flow rate and retention times decreased with decreasing inner diameter, whereas pressure, sensitivity and peak-width increased. Pore width had negligible effects on the chromatographic separation of pigments under the test conditions. Only with acetonitrile as eluent were all the taxonomically important pigments resolved adequately: zeaxanthin (Cyanophyceae), lutein (Chlorophyceae), fucoxanthin (Bacillariopyceae), alloxanthin (Cryptophyceae), peridinin (Dinophyceae).
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  • 42
    ISSN: 1573-1561
    Keywords: Arctiidae ; Lepidoptera ; lichens ; lichen compounds ; HPLC ; sequestration ; chemical defense
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A survey for the presence of sequestered lichen compounds in 103 wild-caught imagines representing eight different genera and 16 different species of the Arctiidae was conducted. Known lichen compounds were detected for the first time in 24 of the analyzed specimens (representing five different genera and 11 different species) based on their HPLC retention times and on their UV-absorption spectra. The anthraquinone parietin, the depside atranorin, as well as a hydrolytic cleavage product of the latter were among the lichen compounds most frequently detected in wild-caught imagines. The observed variation of sequestered lichen compounds in wild-caught imagines with unknown feeding history may be due to several reasons. Lack of lichen compounds in imagines may have been caused, for example, by larvae feeding on lichens with no or only minute amounts of phenolic products. The age of the specimens analyzed may also influence the results obtained. Avoidance of lichen compounds by selective feeding on those parts of lichen thalli that have no or little lichen products may be another reason for the lack of lichen compounds in imagines. Preliminary feeding experiments conducted with larvae ofEilema complana, for example, indicated that the larvae fed exclusively on the algal layer and cortex of the lichenCladonia pyxidata, whereas the medulla, which is rich in fumarprotocetrarie acid, was avoided. As expected, imagines hatching from the larvae were free of this lichen compound. Any ecological role of the sequestered lichen compounds for the herbivores is unknown. It is possible, however, that sequestered lichen compounds may be utilized for the chemical defense of arctiid moths or against microbial pathogens.
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  • 43
    ISSN: 0739-4462
    Keywords: juvenile hormone kinase ; tritiation ; HPLC ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Juvenile hormone epoxide hydrolase (JHEH) and juvenile hormone diol phosphotransferase (JHDPT) were characterized from the Malpighian tubules of day 1 fifth instar Manduca sexta. An improved RP-HPLC assay is described for the major metabolites of (10R, 11S) juvenile hormone I: diol, acid, aciddiol, and diol-phosphate. JHEH is strictly associated with membrane fractions, while JHDPT is cytosolic. JHEH may be solubilized in active form by the nonionic detergents Thesit or MEGA-8. Separation of Malpighian tubule cytosol proteins using preparative isoelectric focusing yields two zones which contain JHDPT activity, at pl 4.8-5.1 and 6.8-8.2. The partially purified JHDPT from either zone requires both ATP and Mg2+ for activity, so this enzyme may be formally called either ATP:juvenile hormone diol phosphotransferase or juvenile hormone diol kinase (EC 2.1.7.3.). Metabolites more polar than JH I aciddiol and JH I diol-phosphate are generated in vivo from either [3H]JH I or [3H]JH I diol. © 1995 Wiley-Liss, Inc.
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    Archives of Insect Biochemistry and Physiology 29 (1995), S. 11-23 
    ISSN: 0739-4462
    Keywords: cricket ; HPLC ; neuropeptide ; diuretic ; antidiuretic ; Malpighian tubule ; fluid secretion ; Mas-DP1 ; achetakinin ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In the Malpighian tubules of Acheta, the distal and middle segments are functionally and morphologically quite distinct (Spring and Kim, Mol Comp Physiol 12:130-145, 1993). Furthermore, they respond quite differently to corpora cardiaca (CC) homogenates, dibutyryl cAMP, and A23187 (Kim and Spring, J Insect Physiol 38:373-381, 1992). In this study we compared secretion by these two regions in response to Acheta and Romalea CC extracts, synthetic Manduca sexta diuretic peptide (Mas-DP1), and the family of synthetic myotropic peptides, the achetakinins, isolated from Acheta. Both Acheta and Romalea CC extracts had opposite effects on the two regions: mid-tubule secretion increased 3-fold whereas secretion by the distal segment declined 75-80%. Mas-DP1 increased secretion by the mid-tubule more than 3-fold and had no effect on the distal segment. All of the achetakinins decreased secretion by the distal tubule, with achetakinin 1 being least effective (55% inhibition) and achetakinin 5 being most effective (75% inhibition). Achetakinins 1 and 2 increased midtubule secretion by 3.7- and 3.3-fold, respectively, whereas the others had no effect on this region. Regarding HPLC fractions of CC extracts, in general the more hydrophilic fractions inhibited secretion by both distal and mid-tubules. The more hydrophobic fractions were nearly uniformly stimulatory when applied to the mid-tubule, and either inhibited secretion or had no effect on the distal region. The possible interpretations of these data and the implications towards future research are discussed. © 1995 Wiley-Liss, Inc.
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  • 45
    ISSN: 1572-8773
    Keywords: HPLC ; pseudobactin ; Pseudomonas fluorescens ; siderophores
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Several iron binding metabolites (siderophores) of Pseudomonas fluorescens B10 (JL-3133) have been detected using C18 reverse phase HPLC coupled with photodiode array detection methods. This analysis utilized a volatile mobile phase of 90% 20 mm NH4HCO3/10% MeOH, pH 6.5. It has been shown to be applicable to other P. fluorescens strains for the detection of related metabolites. Direct scale-up of the analytical HPLC conditions allowed for the efficient preparative isolation of pseudobactin, the principle siderophore produced by P. fluorescens B10 (JL-3133).
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  • 46
    ISSN: 1572-8773
    Keywords: 2,3-dihydroxybenzoylserine ; enterobactin ; Escherichia coli ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Reversed-phase HPLC separation of enterobactin and its 2,3-dihydroxybenzoylserine derivatives was used for a comparative analysis of mutants of Escherichia coli, defective in the regulation of enterobactin biosynthesis (fur), enterobactin transport (fepA) and enterobactin esterase (fes). A complete separation of all 2,3-dihydroxybenzoylserine compounds was achieved: the monomer (DHBS), the linear dimer (DHBS)2 and trimer (DHBS)3, the cyclic trimer, enterobactin, as well as 2,3-dihydroxybenzoic acid. The production of all these compounds was followed after ethylacetate extraction from acidified culture fluids. Enterobactin was found to be the predominant product in all mutant strains. The mutant strains behaved differently with regard to the breakdown products. All degradation products, such as DHBS, (DHBS)2 and (DHBS)3, were detected in the overproducing fur mutant where both transport and esterase are still functioning, while only the monomer, DHBS, was detected in the fepA mutant and no degradation was found in the esterase-deficient fes mutant. From the pattern of breakdown products it may be inferred that the esterase acts in two different ways, depending on whether transport is functioning or not. Thus, esterolytic cleavage of ferric enterobactin after entering the cells results in a mixture of all three hydrolysis products, i.e. DHBS, (DHBS)2 and (DHBS)3, while cleavage of iron-free enterobactin subsequent to its biosynthesis yields only the monomer. Thus, the results of quantitative HPLC analysis of enterobactin and its breakdown products show that different enterobactin esterase products arise, depending on whether iron is bound to enterobactin or not.
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  • 47
    ISSN: 1420-9055
    Keywords: Trophic relationships ; organic biomarkers ; herbivorous zooplankton ; phytoplanktonic pigments ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phyto-zooplankton trophic relationships were studied using phytoplanktonic pigments (chlorophylls and carotenoids) as organic natural markers. Pigments were separated by high-pressure liquid chromatography (HPLC). Comparison of pigment profiles from monospecific cultures of various taxonomic groups (Chlorophyceae, Bacillariophyceae and Cyanobacteria) and from Cladocera crustaceans (Daphnia magna) fed with these cultures, showed that the characteristic pigment associations of the different taxa are conserved during their transfer from primary producers to secondary consumers. Chromatographic profiles of the Bacillariophyceae and Chlorophyceae type were obtained fromDaphnia respectively fed with mixtures of a Chlorophyceae and a diatom species and mixture of a Chlorophyceae and a Cyanobacterium. This showed the importance of this method in demonstrating a possible selective feeding by the herbivorous zooplankton. The observation of pigment profiles of the Dinophyceae type following feeding of a zooplankton assemblage from Lake Pavin within this natural medium (phytoplankton dominated by a Dinophyceae) and of a Chlorophyceae type profile as the same assemblage was fed in the laboratory on phytoplankton from Lake Villerest (composed of about 80% Cyanobacteria and 20% Chlorophyceae), suggested that this method could be applied to the natural environment.
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  • 48
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    Cytotechnology 14 (1994), S. 123-128 
    ISSN: 1573-0778
    Keywords: HPLC ; mammalian cells ; metabolism ; pyrrolidone carboxylic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We have developed a simple and accurate isocratic HPLC method, without any prederivatisation, for the determination of glucose, lactate, glutamine, glutamate, pyrrolidone carboxylic acid and alanine in samples from mammalian cell cultures. The method has been successfully validated with enzyme analysis for each of the compounds. Quantification of pyrrolidone carboxylic acid makes the correction for glutamine decrease due to chemical decomposition very simple and accurate, and avoids some possibly erroneous calculations.
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  • 49
    ISSN: 1573-5044
    Keywords: Catharanthus roseus ; enzyme ; HMG-CoA ; HPLC ; metabolism ; mevalonate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) is an important intermediate in various metabolic pathways, e.g. sterol biosynthesis, ketogenesis and leucine catabolism. The reactions and enzymes involved in the metabolism of HMG-CoA are briefly reviewed. These enzymes have been studied in Catharanthus roseus, a model system for studies on the regulation of secondary metabolic pathways, particularly those leading to terpenoidindole alkaloids. By using HPLC, three HMG-CoA catabolizing enzyme activities have been detected in protein extracts from suspension cultured C. roseus cells: HMG-CoA lyase, 3′-nucleotidase and (tentatively identified) 3-methylglutaconyl-CoA hydratase (HMG-CoA hydrolyase). The enzymes have been partially purified. HMG-CoA is formed from three molecules of acetyl-CoA, via reactions which are catalyzed by two (as in yeast and animal cells, via intermediacy of acetoacetyl-CoA) or by just one enzyme (as in e.g. radish). It is yet not clear which process occurs in C. roseus.
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  • 50
    ISSN: 1573-6881
    Keywords: Chloroplast ATPase ; reaction rate ; HPLC ; Mg2+
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract The influences of total magnesium ion concentration at different total ATP concentrations, and of total ATP concentration, for different total magnesium ion concentrations, on the enzymatic rate of the isolated chloroplast F1 ATPase, have been followed by a chromatographic method consisting in the separation and determination of ADP. From the various series of curves, it is concluded that the experimental results (position of the maxima,K m values) are better fitted by a mechanism involving the activation of the enzyme by magnesium ion and hydrolysis of free ATP, rather than by the classical mechanism, for which the enzyme hydrolyzes the MgATP complex and is inhibited by Mg2+. Although the equations giving the reaction rate are similar in the two cases, the calculated values ofK m are widely different. The value obtained from the classical mechanism does not agree withK D , the dissociation constant of the enzyme-substrate complex, measured by the Hummel and Dreyer method. Moreover, when the total ATP concentration tends toward the total magnesium ion concentration, the nucleotide binding to the enzyme tends toward zero, although it should be maximum if MgATP were the true substrate. Finally, the inhibitory effect of Na+ is more easily explained as a competition between this ion and the activating Mg2+, than by the classical mechanism.
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  • 51
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    Journal of chemical ecology 20 (1994), S. 957-967 
    ISSN: 1573-1561
    Keywords: Chenopodium album ; lambsquarters ; decomposition ; inhibition ; HPLC ; paper chromatography ; phenolics ; radish ; solvent extraction ; ultrafiltration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Aqueous extract of air-dried lambsquarters (Chenopodium album) at 25 mg/ml significantly inhibited germination and growth of radish and wheat seeds. Soybean seed germination was not inhibited; however, hypocotyl growth was significantly reduced. Germination of radish seeds in sand amended with pulverized lambsquarters shoots at 2 and 4 mg/g was reduced 40 and 95%, respectively. Shoot dry weight and plant height were also reduced 30 and 9%, respectively, at 4 mg/g, but not at 2 mg/g concentration. Residues after extraction with water incorporated in sand were not inhibitory, indicating water solubility of the inhibitor(s). Aqueous extract of shoots decomposed for five days lost nearly 40% of its inhibitory effect; 20% of it still persisted in the extract of shoots decomposed for 30 days. The filtrate from ultrafiltration of aqueous extract through a pad of molecular-weight cutoff 1000 inhibited radish seeds germination and growth, indicating that the molecular weight of the inhibitor(s) was less than 1000. Partitioning of the aqueous extract by a series of solvents resulted in isolation of an inhibitor(s) in the butanol fraction. Seven phenolics were identified in this fraction using high-performance liquid chromatography (HPLC). Paper chromatographic analysis of the butanol fraction revealed six bands, of which one band withR f =0.83 inhibited germination and growth of radish seeds. Chlorogenic acid identified by HPLC appeared to be the principal component of the phytotoxin.
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  • 52
    ISSN: 1573-1561
    Keywords: Apiaceae ; Peucedanum ; Lepidoptera ; Noctuidae ; Spodoptera littoralis ; HPLC ; preparative isolation ; furocoumarins ; furanocoumarins ; pyranocoumarins ; growth inhibition ; dietary utilization ; plant chemical diversity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Peucedanum arenarium Waldst. & Kit.,P. austriacum (Jacq.) Koch,P. coriaceum Reichenb.,P. longifolium Waldst. & Kit,P. officinale L.,P. oreoselinum (L.) Moench,P. ostruthium L., andP. palustre (L.) Moench accumulate different structural types of coumarins including simple coumarins, linear furanocoumarins, linear dihydropyranocoumarins, angular dihydrofuranocoumarins and angular dihydropyranocoumarins. Linear furanocoumarins, known for various biological activities, include some well-known antifeedants, such as bergapten, isopimpinellin, and xanthotoxin. The aim of this investigation was to screen the diverse coumarins fromPeucedanum for insecticidal activity. LC was used to analyze and isolate coumarins for the bioassays. A growth inhibition bioassay with 17 derivatives, comprising all structural types fromPeucedanum, carried out withSpodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) as test organism, indicated the majority of the linear furanocoumarins and the angular dihydrofuranocoumarin athamantin as active compounds. Oxygenation of the prenyl residue of linear furanocoumarins decreased activity. Further formation of an ester with angelic acid even resulted in complete inactivity. Five active linear furanocoumarins, bergapten, isopimpinellin, xanthotoxin, isoimperatorin, and imperatorin, and two linear furanocoumarins with a substituted furan ring, peucedanin and 8-methoxypeucedanin, were compared in a dietary utilization bioassay. Relative growth rate (RGR) and relative consumption rate (RCR) divided the tested coumarins in three groups of similar activity. Isopimpinellin and peucedanin slightly decreased RGR and RCR of the treated larvae, and xanthotoxin, isoimperatorin, and 8-methoxypeucedanin heavily decreased RGR and RCR. Bergapten and imperatorin differed by the lowest RGR values and rather high RCR values. The effects caused by these two coumarins indicate specific postingestive toxicity. The results obtained in this study add to the reputation of coumarins to be an effective chemical defense, postulating that chemical diversity is a necessary trait for well-defended plants.
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  • 53
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    Photosynthesis research 41 (1994), S. 157-164 
    ISSN: 1573-5079
    Keywords: bacterial pigments ; bacteriochlorophyll homologues ; chlorobiaceae ; HPLC ; pigment analyses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A reversed-phase High Performance Liquid Cromatography (HPLC) method has been developed to accurately separate bacteriochlorophyllsc, d ande homologues in a reasonably short run time of 60 minutes. By using this method, two well-defined groups of bacteriochlorophyll homologue peaks can be discriminated. The first one consists of 4 peaks (min 24 to 30), which corresponds to the four main farnesyl homologues. The second peak subset is formed by a cluster of up to 10 minor peaks (min 33 to 40). These peaks can be related with series of several alcohol esters of the different chlorosome chlorophylls. The number of homologues was, however, quite variable depending on both, the bacteriochlorophyll and the bacterial species. The method hereby described, also provides a good separation of other photosynthetic pigments, either bacterial (Bacteriochlorophylla, chlorobactene, isorenieratene and okenone) or algal ones (Chlorophylla, Pheophytina and β-carotene). A preliminary screening of the homologue composition of several green photosynthetic bacterial species and isolates, has revealed different relative quantitative patterns. These differences seem to be related to physiological aspects rather than to taxonomic ones. The application of the method to the study of natural populations avoids the typical drawbacks on the pigment identification of overlapping eukaryotic and prokaryotic phototrophic microorganisms, giving further information about their physiological status.
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  • 54
    ISSN: 1573-5079
    Keywords: cyanobacteria ; HPLC ; monomer ; Photosystem I ; state transitions ; trimer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In cyanobacteria, solubilization of thylakoid membranes by detergents yields both monomeric and trimeric Photosystem I (PS I) complexes in variable amounts. We present evidence for the existence of both monomeric and trimeric PS I in cyanobacterial thylakoid membranes with the oligomeric state depending ‘in vitro’ on the ion concentration. At low salt concentrations (i.e.≤10 mM MgSO4) PS I is mainly extracted as a trimer from these membranes and at high salt concentrations (i.e.≥150 mM MgSO4) nearly exclusively as a monomer, irrespective of the type of salt used (i.e. mono- or bivalent ions) and the temperature (i.e. 4°C or 20°C). Once solubilized, the PS I trimer is stable over a wide range of ion concentrations (i.e. beyond 0.5 M). A model is presented which suggests a monomer-oligomer equilibrium of PS I, but also of PS II and the cyt. b6/f-complex in the cyanobacterial thylakoid membrane. The possible physiological role of this equilibrium in the regulation of state transitions is discussed.
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  • 55
    ISSN: 1573-5133
    Keywords: Fish ; Displacement ; Home range ; Natural marking ; Radioactivity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Synopsis On the basis that γ-emitting artificial radionuclides, present in nuclear plant radioactive wastes, can be considered as ‘natural’ collective markers of fish living downstream from a radioactive discharge, we studied the restricted movements of chub in a reservoir on the Lower Rhône river where the Marcoule nuclear plant is located. A qualitative determination, based on the detection of specific radioelements in our samples originating from radioactive waste, and a quantitative determination of the cesium-137 concentration in the samples, were used to identify fish radioactively marked by the effluent. Individual measurements of γ-radioactivity in 49 adult chub captured at two stations, each 2 km long and 6 km apart, divided the fish into two distinct sub-units: one living downstream of the discharge pipe, in which 73% were marked, and the other upstream, in which 79% were not marked. Similar results were found in two neighbouring stations when we analysed previous radioecological measurements of γ-radioactivity in groups of chub. After combining all the data concerning chub, detailed information was obtained on the spatial stability of the chub population in the reservoir. Two spatially different stocks were found and each stock can be divided in two components: a sedentary component that remains in a restricted zone (its home range), and a mobile component that undertakes movements between the two zones.
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  • 56
    ISSN: 0749-503X
    Keywords: Cell wall ; glucan ; chitin ; killer toxin ; HPLC ; S. cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have previously shown that mutations in the yeast KNR4 gene resulted in pleiotropic cell wall defects, including resistance to killer 9 toxin, elevated osmotic sensitivity to SDS and increased resistance to zymolyase, a (1→3)-β-glucanase. In this report, we further demonstrated that knr4 mutant cells were more permeable to a chromogenic substrate, X-GAL, suggesting that the mutant cell walls were leakier to certain non-permeable molecules. To determine if these defects resulted from structural changes in the cell walls, we analysed the alkali-insoluble cell wall components using HPLC assays developed for this purpose. Comparative analysis using four isogenic strains from a ‘knr4 disrupted’ tetrad demonstrated that mutant cell walls contained much less (1→3)-β-glucan and (1→6)-β-glucan; however, the level of chitin, a minor cell wall component, was found to be five times higher in the mutant strains compared to the wild-type strains. The data suggested that the knr4 mutant cell walls were dramatically weakened, which may explain the pleiotropic cell wall defects.
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  • 57
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    Archives of Insect Biochemistry and Physiology 27 (1994), S. 27-38 
    ISSN: 0739-4462
    Keywords: hormone ; HPLC ; ovary ; antibody ; PAGE ; pulse-chase ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Trypsin modulating oostatic factor (TMOF) was followed by RIA in the ovary of female Aedes aegypti before and after the blood meal. The amount of TMOF in a pair of ovaries from females fed sugar for 3 days or blood for 24 h was low (1.7 ng). Between 24 and 48 h after the blood meal the amount of TMOF in the ovaries rapidly increased and reached a peak of 104 ng at 48 h. The amount of TMOF in the head of a female A. aegypti was very low (0.05 to 0.1 ng) during sugar and blood feeding. Immunocytochemical methodology identified the follicular epithelium as the site of biosynthesis of TMOF in the ovary. Females ovariectomized and fed a blood meal continued to synthesize trypsin for 64 h, whereas intact controls stopped at 40 h, indicating that a factor from the ovary regulates trypsin biosynthesis. Ovaries incubated in vitro with [3H]proline synthesized [pro-3H]TMOF that was identified by HPLC and by anti-TMOF serum. The ovary started to synthesize TMOF in vitro 24 h after the blood meal, and the synthesis reached a peak at 36 h and then declined. The synthesis of TMOF by the ovary is closely correlated with the termination of trypsin biosynthesis in the female mosquito's midgut. Ovaries that were pulsed with [3H]proline for 30 min synthesized [pro-3H]TMOF which was chased into the medium with unlabeled proline, indicating that the hormone is secreted by the ovary. These results indicate that TMOF is a secretory peptide, synthesized by the ovarian follicular epithelium and that it modulates trypsin biosynthesis in the mosquito's gut. © 1994 Wiley-Liss, Inc.
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  • 58
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    Archives of Insect Biochemistry and Physiology 27 (1994), S. 11-25 
    ISSN: 0739-4462
    Keywords: hormone ; HPLC ; gas chromatography ; tissue culture ; methoxyhydrin derivative ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Synthesis of (10R)-juvenile hormone III (JH III) outside the corpora allata (CA) was investigated in female Aedes aegypti. Intact females or ligated abdomens of blood-fed and sugar-fed females synthesized in vivo [12-3H]JH III-like molecules from [12-3H]-methyl farnesoate, indicating that an organ(s) in the female abdomen, other than the CA, converted methyl farnesoate into JH III. To find out the organ(s) that synthesized JH III-like molecules, ovaries, fat bodies, and midguts were incubated in vitro with [12-3H]methyl farnesoate and the synthesis of JH III-like molecules was compared with JH III synthesized by CA. To identify tissue(s) having both farnesoic acid methyl transferase and farnesoate epoxidase, enzymes that convert farnesoic acid into JH III, ovaries, and fat bodies were removed from sugar and blood-fed females and incubated with [12-3H]farnesoic acid. Chemical derivatization by methoxyhydrin formation followed by esterification with (+)-α-methoxy- α-trifluoromethyl phenylacetic (MTPA) acid chloride and reversed phase liquid chromatography identified (10R)-JH III methoxyhydrin (+)-MTPA ester as the sole JH III-like molecule produced in tissue culture incubation of ovaries. Since only (10R)-JH III is produced and not racemic JH III, the oxidation of farnesoic acid must be enzymatically mediated. Ovaries and corpora allata of female A. aegypti also synthesized [3H,14C]JH III from L-[methyl-3H]methionine and [14C]acetate which was characterized by HPLC and gas chromatography. These results suggest that mosquito ovary can synthesize (10R)-JH III from farnesoic acid, and that this tissue synthesizes JH III-like molecules from L-methionine and acetate. © 1994 Wiley-Liss, Inc.
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  • 59
    ISSN: 1432-1351
    Keywords: Photoreception ; Extraretinal Photoreceptor ; Chromophore ; Opsin ; Reptile ; Immunocytochemistry ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Since the beginning of this century evidence has accumulated which demonstrates that non-mammalian vertebrates possess photoreceptors situated deep within the brain. While many attempts have been made to localize these sensory cells, studies have either failed or been inconclusive. In this report we have used several experimental approaches to localize the deep brain photoreceptors of the lizard Anolis carolinensis. Using 3 antibodies that bind vertebrate cone opsins, we have immunolabelled cerebrospinal fluid (CSF)-contacting neurons located at the ventricular border within the nucleus ventromedialis of the septum. Western blot analysis indicates that these antibodies recognized a single 40 kD protein in ocular, anterior brain, and pineal extracts. Immunoblots of rodent brain did not show a similar protein band. We have also identified specific retinoids associated with phototransduction (11-cis and all-trans-3,4-didehydroretinaldehyde) within anterior brain extracts. This combined data provides the most detailed analysis of deep brain photoreceptors in any vertebrate. Consequently, we feel Anolis provides an excellent model to study this unexplored sensory system of the vertebrates.
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  • 60
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    Mycopathologia 121 (1993), S. 179-192 
    ISSN: 1573-0832
    Keywords: ELISA ; Fusaria ; HPLC ; Mycotoxins ; TLC ; Trichothecenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The production of type A trichothecene mycotoxins by 19 Fusaria, including 12Fusarium sporotrichioides, 4F. chlamydosporum and 3F. graminearum at 15°C and 25°C over a 35-day period was analyzed by ELISA using antibodies cross-reactive with most type A trichothecenes after conversion to T-2 tetraol tetraacetate. The toxin production peaked at 20–25 days of incubation with maximum yield between 4–6 mg type A trichothecene/ml of culture medium for 5F. sporotrichioides cultures and between 1 to 2 mg/ml for 6F. sporotrichioides cultures. OneF. sporotrichioides produced 700 µg type A trichothecenes/ml of culture medium. Detectable type A trichothecene was also found in the culture extracts ofF. chlamydosporum andF. graminearum, but the yield was very low (less than 100 µg/ml). Quantitative determination of individual trichothecenes was achieved by separation of different toxin in HPLC and followed by ELISA analysis. Eight to 10 immunoreactive peaks, corresponding to various type A trichothecenes, were detected in all the fungal extracts. T-2 tetraol (T-2-4ol), 4-acetyl-T-2 tetraol (4-Ac-T-2-4ol), neosolaniol (NEOS), diacetoxyscirpenol (DAS), HT-2 and T-2 toxin accounted for more than 85% of the total toxins. In general, low temperature was preferred for total type A trichothecene production. More T-2-4ol, 4-Ac-T-2-4ol, HT-2 and DAS were produced at 25°C. In contrast, more T-2 toxin and NEOS were produced at 15°C. Transformation of T-2 toxin and NEOS to polar metabolites such as T-2-4ol, 4-acetyl-T-2-4ol and HT-2 by various strains were observed at both temperatures after 25 days incubation.
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  • 61
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    Mycopathologia 121 (1993), S. 27-32 
    ISSN: 1573-0832
    Keywords: Corn ; Fusarium graminearum ; HPLC ; Zearalenone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A high pressure liquid chromatographic (HPLC) method to determine zearalenone in corn contaminated withFusarium graminearum is described. After extraction with methanol-water and solvent partition, samples were cleaned up by applying the extract to a disposable silica cartridge and by eluting the toxin with a mixture of hexane/dry ethyl ether (5/5). Separation was achieved by a reverse phase μBondapak C18 column followed by fluorescence detection using an excitation wavelength at 274 nm and an emission wavelength at 440 nm. Detection limit was about 5 ng. Recoveries ranging from 85.37 to 100.97%, in standard solutions range 30–0.5 µg/ml, were found.
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  • 62
    ISSN: 1420-9055
    Keywords: chlorophylla ; chlorophyllidea ; pheopigments ; spectrophotometry ; HPLC ; marine sediments
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pigment concentrations (chlorophylla, chlorophyllidea and pheopigmentsa) were measured by HPLC and spectrophotometry with acidification on 57 samples collected in different marine coastal sediments, containing autochthonous microphytes, and with various organic matter contents (plant detritus, biodeposits or hydrocarbons). Statistical analysis shows that the spectrophotometry with acidification, as compared to HPLC, gives reliable values for chlorophylla. Chlorophyllidea concentrations may be considered as negligible. Though spectrophotometric methods are sometimes questioned when applied to sediments they appear to give easy, quick and good estimates of Chla contents in benthic microphytes for hydrobiological studies in coastal areas.
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  • 63
    ISSN: 1432-203X
    Keywords: CAT ; GUS ; HPLC ; Co-transformation ; Internal Standard ; Tobacco Protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The use of transient gene expression assays for the study of natural or engineered plant promoters is affected by a considerable degree of inter-experiment variability. As a means of obtaining interpretable data from a limited number of experiments, we worked out conditions for the simultaneous determi nation of the activity of two reporter genes, a “sample” and a “reference”, ona single extract of co-transformed protoplasts. ß-glucuronidase (GUS) and chloramphenicol acetyl transferase (CAT) genes, both under the control of the CaMV 35S promoter, were transferred into tobacco (Nicotiana tabacum L.) protoplasts on two independent plasmids. The parallel expression of the two reporter genes in several independent co-transformation experiments was verified. Conditions for the use of a single protoplast extraction buffer and for the simultaneous assay of both reporter gene activities were set up. A HPLC method for the non-radioactive determination of both enzyme activities on a single aliquot of the reaction mixture was developed. The resulting procedure was tested using the GUS gene as “reference” and the CAT gene, under the control of either wild type or upstream-deleted (−90) CaMV 35S promoter, as “sample”. The protocol is simple and allows the fast analysis of plant promoters in the presence of a true internal standard under conditions in which assay manipulations are reduced to a minimum and both reporter gene activities are subjected to the same experimental treatments.
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  • 64
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    Journal of chemical ecology 19 (1993), S. 395-410 
    ISSN: 1573-1561
    Keywords: Oreina gloriosa ; Coleoptera ; Chrysomelidae ; chemical defense ; cardenolides ; quantitative variation ; aging ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The defensive secretion of the alpine chrysomelidOreina gloriosa is a complex mixture of mainly cardenolides and tyrosine betaine. Individually sampled secretions of adult laboratory-reared and field-collected beetles were analyzed by reverse-phase HPLC; 16 secretion components were quantified. Quantities and concentrations of different components were significantly affected by the age, sex, and reproductive status of individual beetles. Aging was correlated with marked increases (up to 4.4-fold) and decreases (up to 2.7-fold) of quantities and concentrations of several components. Differences between the sexes were smaller, but quantities of all components and concentrations of several components were larger in laboratory-reared females than in males. There was less of one component of the secretion in mated than unmated females, but the concentrations of four secretion components were higher (up to 1.6-fold) in mated females.
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  • 65
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    The journal of membrane biology 136 (1993), S. 281-288 
    ISSN: 1432-1424
    Keywords: Thiamine triphosphate ; Anion channel ; Oxythiamine ; Neuroblastoma ; Patch clamp ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract In neuroblastoma cells, the intracellular thiamine triphosphate (TTP) concentration was found to be about 0.5 μ m, which is several times above the amount of cultured neurons or glial cells. In inside-out patches, addition of TTP (1 or 10) μ m to the bath activated an anion channel of large unit conductance (350–400 pS) in symmetrical 150 mm NaCl solution. The activation occurred after a delay of about 4 min and was not reversed when TTP was washed out. A possible explanation is that the channel has been irreversibly phosphorylated by TTP. The channel open probability (P o) shows a bell-shaped behavior as a function of pipette potential (V p). P o is maximal for −25 mV〈V p〈10 mV and steeply decreases outside this potential range. From reversal potentials, permeability ratios of PCl/ PNa = 20 and PCl/Pgluconate = 3 were estimated. ATP (5 mm) at the cytoplasmic side of the channel decreased the mean single channel conductance by about 50%, but thiamine derivatives did not affect unit conductance; 4,4′ -diisothiocyanostilbene-2,2′-disulfonic acid (0.1 mm) increased the flickering of the channel between the open and closed state, finally leading to its closure. Addition of oxythiamine (1 mm), a thiamine antimetabolite, to the pipette filling solution potentiates the time-dependent inactivation of the channel at V p=−20 mV but had the opposite effect at +30 mV. This finding corresponds to a shift of P o towards more negative resting membrane potentials. These observations agree with our previous results showing a modulation of chloride permeability by thiamine derivatives in membrane vesicles from rat brain.
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  • 66
    ISSN: 1432-0878
    Keywords: Pineal organ ; Retina ; Photoreceptors ; Photopigment ; Immunocytochemistry ; HPLC ; Autoradiography ; Mouse (C57BL)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The aim of the present study was to characterize the rod-opsin immunoreaction in the mammalian pineal organ. Pigmented mice (strain C57BL) were selected as the animal model. Immunocytochemical investigations involving the use of highly specific polyclonal and monoclonal antibodies against bovine rod-opsin (the apoprotein of the photopigment rhodopsin) showed that approximately 25% of all pinealocytes were rod-opsin immunoreactive. Immunoblotting techniques revealed three protein bands of approximately 40, 75, and 110 kDa; these were detected by the monoclonal antibody and the polyclonal antiserum in retinal and pineal extracts. These protein bands presumably represented the monomeric, dimeric and trimeric forms of rod-opsin. The amount of rod-opsin in retina and pineal organ was quantified by means of an enzyme-linked immunosorbent assay. This yielded 570±30 pmoles rod-opsin per eye and 0.3±0.05 pmoles rod-opsin per pineal organ. High pressure liquid chromatography analysis of whole eye extracts demonstrated the chromophoric group of the photopigment rhodopsin, 11-cis retinal, and its isomer, all-trans-retinal. A shift from 11-cis retinal to all-trans-retinal was found upon light adaptation. No retinals were detected in the pineal organ. Autoradiographic investigations showed that 3H-retinol, intraperitoneally injected into the animals, was incorporated into the outer and inner segments of retinal photoreceptors, but not into the pineal organ. It is concluded that the mouse pineal organ contains the authentic apoprotein of rhodopsin but that it lacks retinal derivatives as essential components of all known vertebrate photopigments. Consequently, the “photoreceptor-specific” proteins of the mammalian pineal organ are not involved in photoreception and phototransduction, but may serve other functions to be explored in future studies.
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  • 67
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    Journal of applied phycology 5 (1993), S. 623-628 
    ISSN: 1573-5176
    Keywords: -carotene ; chlorophylla ; cyanobacterial mats ; HPLC ; myxoxanthophyll
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The efficiency of 9:1 acetone-water, DMSO and boiling 9:1 ethanol-water in extracting chlorophyll and carotenoid pigments from benthic cyanobacterial mats from Antarctica for HPLC (high performance liquid chromatography) analysis was examined. Considerable breakdown of chlorophylla was observed after 5 min extraction in boiling ethanol and 2 h extraction in DMSO. Over 50% of the chlorophylla was degraded to chlorophyllidea and there was substantial loss of carotenoids after a 15 h exposure of ground cells to cold 9:1 acetone-water. Mild sonication of ground mat material in 9:1 acetone-water followed by a 4 h extraction at 4 ° C was found to minimise chlorophylla breakdown and dramatically improved the extraction efficiency of chlorophylla, myxoxanthophyll and -carotene.
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  • 68
    ISSN: 1573-5079
    Keywords: carotenoids ; chlorophylls ; HPLC ; phase partition ; xanthophylls
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    Topics: Biology
    Notes: Abstract Thylakoid membranes from spinach were fragmented mechanically and separated into vesicles originating from grana and stroma-exposed lamellae (Andreasson et al. (1988) Biochim Biophys Acta 936: 339–350). The grana vesicles were further fragmented and separated into smaller vesicles originating from different parts of the grana (Svensson and Albertsson (1989) Photosynth Res 20: 249–259). All vesicles so obtained were analyzed with respect to chlorophyll and carotenoid composition by reverse phase HPLC. For all fractions the following relations (mole/mole) were found: 1 carotenoid per 4 chlorophyll (a+b), 2 lutein per 5 chlorophyll b and 5 violaxanthin per 100 chlorophyll (a + b). The contents of lutein and neoxanthin were each linearly related to chlorophyll b and β-carotene was linearly related to chlorophyll a.
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  • 69
    ISSN: 1573-5168
    Keywords: GnRH ; salmon GnRH ; chicken GnRH-II ; HPLC ; RIA ; bluefin tuna ; red seabream ; black seabream ; red spotted grouper ; Japanese flounder
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Brain extracts from bluefin tuna, Thunnus thynnus, red seabream, Pagrus major, black seabream, Acanthopagrus schlegeli, red spotted grouper, Epinephelus akaara and Japanese flounder, Paralichthys olivaceus, were analyzed by high performance liquid chromatography (HPLC) and specific radioimmunoassays. Immunoreactive material co-eluting from HPLC with salmon gonadotropin-releasing hormone (GnRH) and chicken GnRH-II, respectively, was found in all five species. In addition, a GnRH immunoreactive fraction showing the same HPLC retention time as lamprey GnRH-I was detected in the brain extracts of all species examined when using an unspecific radioimmunoassay which detects several GnRH forms, including lamprey GnRH-I. In the Japanese flounder brain extract, a fourth GnRH immunoreactive fraction was detected with the unspecific radioimmunoassay which did not co-elute with any of the six synthetic GnRH standards used in the present study.
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  • 70
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    Journal of chemical ecology 19 (1993), S. 2231-2244 
    ISSN: 1573-1561
    Keywords: Allelopathy ; allelochemicals ; phytotoxin ; gramine ; hordenine ; HPLC ; TEM ; micrograph ; autophagy ; barley ; Hordeum vulgare ; Sinapis alba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The release of alkaloids by barley was quantified by HPLC. Hordenine was released from the roots of barley in a hydroponic system for up to 60 days. The amount reached a maximum, 2μg/plant/day, at 36 days, then declined. Effects on white mustard by hordenine and gramine included reduction of radicle length and apparent reduction in health and vigor of radicle tips. Transmission electron microscopic examination of white mustard radicle tips exposed to hordenine and gramine showed damage to cell walls, increase in both size and number of vacuoles, autophagy, and disorganization of organelles. The evidence of the morphological and primary effects of barley allelochemicals at the levels released by living plants indicates that the biologically active secondary metabolites of barley may lead to a significant role in selfdefense by the crop.
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  • 71
    ISSN: 1573-1561
    Keywords: Barley ; Hordeum vulgare ; allelochemicals ; hordenine ; gramine ; quantification ; HPLC ; cultivars
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    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A method was devised for the extraction and quantification of hordenine and gramine from barley (Hordeum vulgare) tissue using HPLC techniques. Quantification was by peak area, the relationship between peak area and concentration of authentic standards being linear for both hordenine and gramine. Significant differences in the ability of three lines of barley to produce hordenine and gramine were detected using this method.
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  • 72
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    Plant cell, tissue and organ culture 35 (1993), S. 181-193 
    ISSN: 1573-5044
    Keywords: HPLC ; membrane rafts ; microtubule bioassay ; roots ; Taxol ; Taxus tissue eulture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Callus was induced from Taxus baccata cv. Repandens Parsons ex Rehd., T. brevifolia Nutt., T. cuspidata Sieb. & Zucc., and T. x media cvs. Hicksii and Densiformis Rehd. using different concentrations of 2,4-d-(2,4-dichlorophenoxyacetic acid), IBA (indole-3-butyric acid), or NAA α-naphthalene acetic acid in combination with kinetin. All cultures grew slowly following the first subculture, and a majority turned brown and ceased growth within the next six to twelve months. The callus cultures which lived, continued to grow very slowly for one to two years before the growth rate improved. Initiation of roots and shoot primordia-like structures occurred on some cultures maintained in the dark, and 16 h light/8 h dark, respectively. A fast-growing, habituated callus line (CR-1) derived from T. x media Rehd. cv. Hicksii was established from callus initiated in 1986. Supplementing the medium with casein hydrolysate and both fructose and glucose enhanced the growth rate. A great deal of heterogeneity was found among and within the callus, with respect to the amount of taxol produced. The callus exhibited levels of taxol ranging from 0.1 to 13.1 mg kg-1 (0.0001 to 0.0131%) on a dry weight basis. Overall, the older brown-colored callus produced more taxol than the younger pale yellow-colored callus. The presence of taxol in callus samples was established by high performance liquid chromatography, its biological activity confirmed by a microtubule-stabilizing bioassay and its structure confirmed using one-and two-dimensional 1H and 13C nuclear magnetic resonance spectroscopy.
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  • 73
    ISSN: 1573-5079
    Keywords: green sulfur bacteria ; chlorosome ; bacteriochlorophyll c ; bacteriochlorophyll d ; bacteriochlorophyll e ; HPLC ; absorption spectrum ; mass spectroscopy ; Chlorobiaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The chlorosomal bacteriochlorophyll (BChl) composition of the green sulfur bacteria Chlorobium vibrioforme and Chlorobium phaeovibrioides was investigated by means of normal-phase high-performance liquid chromatography. From both species a number of homologues was isolated, which were identified by absorption and 252Cf-plasma desorption mass spectroscopy. Besides BChl d, C. vibrioforme contained a significant amount of BChl c, which may provide an explanation for the previous observation of at least two spectrally different pools of BChl in the chlorosomes of green sulfur bacteria (Otte et al. 1991). C. phaeovibrioides contained various homologues of BChl e only. Absorption spectra in acetone of BChl c, d and e, as well as bacteriopheophytin e are presented. No systematic differences were found for the various homologues of each pigment. In addition to farnesol, the mass spectra revealed the presence of various minor esterifying alcohols in both species, including phytol, oleol, cetol and 4-undecyl-2-furanmethanol, as well as an alcohol of low molecular mass, which is tentatively assumed to be decenol.
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  • 74
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    Journal of chemical ecology 19 (1993), S. 1691-1701 
    ISSN: 1573-1561
    Keywords: Allelochemicals ; Gliricidia sepium ; Sorghum vulgare ; HPLC ; aglycone ; phenolic acids ; weed control ; root biomass ; shoot biomass ; inflorescence biomass ; grain yield ; weed biomass
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Allelochemicals fromGliricidia sepium were extracted, identified, and quantified using HPLC. Fifteen toxic compounds, namely gallic acid, protocatechuic acid,p-hydroxybenzoic acid, gentisic acid,Β-resorcyclic acid, vanillic acid, syringic acid,p-coumaric acid,m-coumaric acid,o-coumaric acid, ferulic acid, sinapinic acid (trans andcis forms), coumarin, and myricetin were identified and quantified. These compounds from the plant extracts were tested on the seeds of the crop plant,Sorghum vulgare. Rate of germination of the seeds and root elongation were found to be inhibited by the various compounds of the extract. Different quantities ofGliricidia leaf mulch, viz., 400, 800, and 1200 g/m2 applied to theSorghum grown fields, were found to effectively control weeds. Mulching improved the total yield ofSorghum. Leaf manuring and mulching showed better crop yield when applied up to 800 g ofGliricidia leaf/m2. Crop yield was better in mulch-applied fields when compared to the manure-applied ones.
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  • 75
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    Biotechnology and Bioengineering 42 (1993), S. 815-820 
    ISSN: 0006-3592
    Keywords: pravastatin ; ML236B sodium salt ; computer coupled control system ; HPLC ; on-line monitoring ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An automatic feeding process for microbial hydroxylation of ML236B sodium salt (ML-236B Na; compactin) by Streptomyces carbophilus SANK 62585 was developed. The hydroxylated product, pravastatin sodium salt (pravastatin; trade name Mevalotin), is an inhibitor of 3-hydroxy-3-methyglutaryl-coenzyme A reductase (HMG-CoA reductase) used as cholesterol-lowering drug. The hydroxylation activity of S. carbophilus was induced by the addition of ML236B Na to culture broth but inhibited by high concentration of ML236B Na. In order to obtain high conversion yield, it was necessary to maintain optimum ML236B Na concentration throughout the fermentation by continuous feeding. For this purpose, we developed an on-line monitoring method, which mainly consisted of a cross-flow filtration module, high-performance liquid chromatography (HPLC) analyzer, feed pump, and microcomputer for regulation of ML236B Na concentration. An algorithm for control of ML236B Na feed rate based on feedback and feed-forward control where conversion rate after Δt was estimated by using regression analysis of the five latest values of conversion rate. In a fed-batch culture employing this system, the concentration of ML236B Na was maintained at optimum level during the fermentation and the productivity of pravastatin was increased threefold over that obtained in manual control culture. © 1993 John Wiley & Sons, Inc.
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  • 76
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    Archives of Insect Biochemistry and Physiology 24 (1993), S. 1-19 
    ISSN: 0739-4462
    Keywords: chromatography ; cytochrome b5 ; monooxygenase ; HPLC ; immunoelectrophoresis ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A cytochrome P-450 (P-450) was purified from abdominal microsomes of untreated and phenobarbital treated susceptible (S+) and insecticide-resistant (LPR) houseflies using HPLC purification procedures. The purified P-450s exhibited the same apparent molecular masses (54,400 dalton) and could not be distinguished from each other on the basis of HPLC chromatographic properties, reduced CO-difference absorbance maxima (447 ± 0.5 nm), or partial NH2-terminal sequences (MLLLLLLIVVTTLYIFAKL). Since these P-450s were indistinguishable and were immunologically identical to cytochrome P-450lpr we conclude that these P-450s are in fact cytochrome P-450lpr. The level of P-450lpr was increased twofold after PB exposure in the S+ strain, while the level of P-450lpr did not change in the LPR strain. Immunoinhibition of ECOD activity with anti-P-450lpr antiserum suggests that PB treatment in S+ house flies induced P-450lpr, and other forms of cytochromes P-450 with high activity toward this substrate. © 1993 Wiley-Liss, Inc.
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  • 77
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    Plant cell reports 11 (1992), S. 132-136 
    ISSN: 1432-203X
    Keywords: Transformation ; Palaver somniferum ; Agrobacterium rhizogenes ; Morphinan alkaloids ; ELISA ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Transformed cultures of opium poppy (Papaver somniferum L.) were established by infecting hypocotyl segments with Agrobacterium rhizogenes MAFF 03-01724. Undifferentiated calli formed on the infected site grew satisfactorily on phytohormone-free solid medium in the dark and produced opine, mikimopine, which could not be detected in a normal culture. Numerous adventitious shoots developed from transformed calli during subculture. The transformed shoots separated individually were cultured on phytohormone-free MS solid medium at 22 ° C under 14 h/day light. They displayed wider leaves and longer internodes than shoots established from seeds or non-transformed root culture. The content of morphinan alkaloids in the cultures and regenerated shoots were quantitatively analyzed by enzyme-linked immunosorbent assay and high performance liquid chromatography. HPLC analysis revealed that non-transformed shoots contained much more codeine (1310 gmg/g dry wt.) than morphine (50 μg/g dry wt.), while the transformed shoot cultures did not contain morphine, although the level of morphinan alkaloids in the transformed shoots (213 μg morphine equivalents/g fr. wt.) was comparable to that in non-transformed shoots (182 μg morphine equivalents/g fr. wt.) by ELISA.
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  • 78
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    Theoretical and applied genetics 85 (1992), S. 407-414 
    ISSN: 1432-2242
    Keywords: Maize ; Zein ; Prolamins ; Mutants Genes ; HPLC ; Quantitation ; Epistasis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Zeins, the major endosperm proteins in maize (Zea mays L.), are deficient in the essential amino acids lysine and tryptophan. Some mutant genes, like opaque-2 (o2) and floury-2 (fl2), reduce the levels of A- and B-zeins, thereby improving maize's nutritional value. Other mutants, such as amylose-extender (ae), floury-1 (fl1), soft starch (h), dull-1 (du), shrunken-1 (sh1), sugary-1 (su1), sugary-2 (su2), and waxy (wx), primarily affect starch composition, but also alter zein composition. We undertook this study to examine the effects of some of these mutant genes on A/B-zein composition and to study the interactions of these genes in double-mutant combinations. Endosperm prolamins were extracted from inbred B37, ten near-isogenic single mutants (ae, du, fl1, fl2, h, o2, sh1, su1, su2, and wx), and most double-mutant combinations. Zeins in these extracts were fractionated by reversed-phase highperformance liquid chromatography (RP-HPLC) into 22–24 peaks. Of the resulting 22 major peaks the areas of 16 (per milligram endosperm) were significantly affected by individual mutant genes relative to the zein composition of the normal inbred. In combination these genes exhibited significant epistatic interactions in regulating the expression of individual A/B zeins. Epistatic interactions were judged to be significant when the amount of a peak in a double mutant differed from the averages for the peak in the two respective single mutants. The o2 gene, alone and in combination with other mutant genes, significantly decreased the amounts of many individual zeins. The effect of the o2 gene was the greatest of all the genes examined. Various clustering techniques were used to see if mutant effects could be grouped; among these was principal component analysis, a multivariate statistical technique that analyzes all peak sizes simultaneously. Three-dimensional scatter graphs were constructed based on the first three principal components. For the single mutants, these showed no relationships to gene actions; for the double mutants, however, this technique showed that four single mutants, o2, sh1, su1 and su2, had the greatest effects on zein composition when combined with each other and with the remaining six single mutants.
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    Cell & tissue research 270 (1992), S. 601-607 
    ISSN: 1432-0878
    Keywords: Dopamine ; HPLC ; Immunocytochemistry ; Hydra attenuata (Cnidaria)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary High performance liquid chromatography (HPLC), with electrochemical detection, is an analytical method sensitive enough to permit quantification of dopamine, dihydroxyphenylalanine (DOPA) and 5-S-cysteinyl DOPA in single or hemisected specimens ofHydra attenuata. Dopamine and 5-S-cysteinylDOPA appear to be the quantitatively predominant catechol compounds inH. attenuata, whereas DOPA is present in minor amounts. The presence of DOPA and 5-S-cysteinylDOPA, and the quantitative correlation between dopamine and these compounds in many specimens, suggests that dopamine inH. attenuata, as in higher animals, is formed through decarboxylation of DOPA. Contrary to the dopaminergic nerves in higher animals, DOPA inHydra seems to be oxidized and 5-S-cysteinyl DOPA is formed as a by-product. The oxidation of DOPA indicates that the hydroxylation of tyrosine into DOPA in the tissues ofH. attenuata is mediated by a tyrosinase rather than a tyrosine hydroxylase. Immunocytochemical methods demonstrate a highly variable distribution of dopamine in the tissues of different specimens ofH. attenuata. Dopamine immunoreactivity is confined to ectodermal tissue and can be found in several different cell types including nerve cells, battery cells, nematocytes, epithelial cells and interstitial undifferentiated cells. The large amounts of dopamine found in some specimens ofH. attenuata indicate some biological function, although its sporadic occurrence in neurites makes it less plausible as a generally utilized neurotransmitter in this animal.
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  • 80
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    Journal of industrial microbiology and biotechnology 11 (1992), S. 43-51 
    ISSN: 1476-5535
    Keywords: Rhodotorula rubra sp. TP1 ; Pigment characterization ; HPLC ; TLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A new strain ofRhodotorula rubra has been isolated from yogurt which shows promise as a source for pigment and protein feed for aquacultured animals. The pigment was extracted by rupturing the cells with the French press followed by extraction with acetone and purification of the acetone extract using petroleum ether and cold 10% NaCl. The absorption spectrum indicated that the pigment was a carotenoid, the chemistry of which was examined using nuclear magnetic resonance, mass spectroscopy and resonance Raman spectroscopy. A reverse-phase HPLC equipped with octadecylsilylated (ODS) silica column showed nearly 80-times more pigment production under similar cultural conditions thatPhaffia rhodozyma. The isolate grows optimally at 20°C when grown on a variety of media. Its morphology has been studied using transmission electron microscopy, scanning electron microscopy and phase contrast microscopy. From the results of the API system, the isolate was identified asRhodotorula rubra.
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  • 81
    ISSN: 1573-1561
    Keywords: Bracken fern ; Pteridium aquilinum var.Caudatum ; tropics ; ptaquiloside ; pterosin A ; pterosin B ; quantitation ; HPLC ; blade growth ; semiochemicals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A negative correlation has been found between the amounts of pterosins A and B and ptaquiloside per biomass unit, and the growth stage of the blade of bracken. Their concentration decreased progressively from the crozierto the mature frond, where it attained less than 5% of the initial value. The growth was measured following the total blade length, its height, moisture content, and time of emergence from the soil surface. Quantitation of these compounds was achieved by HPLC using a water extraction, methylene chloride treatment, and silica gel microcolumn cleanup sequence. Pterosins were unevenly distributed in the blade, whereas ptaquiloside maintained a constant concentration throughout. Rhizomes contained only minor amounts of these compounds. Their possible role as semiochemicals in bracken is discussed.
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  • 82
    ISSN: 1573-5117
    Keywords: Heterocapsa sp. ; Olisthodiscus luteus ; cycles ; pigments ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photosynthetic pigment composition was studied in batch cultures of Heterocapsa sp. and Olisthodiscus luteus growing exponentially in a 12:12 light:dark cycle. Both species divided in the dark. The synthesis of pigments was continuous for both species. However for chlorophyll c and peridinin, in Heterocapsa sp., and chlorophyll c and fucoxanthin, in O. luteus, (pigments belonging to light harvesting complexes) the synthesis was significantly higher during the light period. Concentrations per total cell volume (TCV) of chlorophyll a, chlorophyll c, peridinin and diadinoxanthin in Heterocapsa sp., and chlorophyll a, chlorophyll c, fucoxanthin and violaxanthin in O. luteus, showed a maximum at the onset of light and decreased during the light period. The values of the chlorophyll a:chlorophyll c, chlorophyll a:peridinin and chlorophyll a:fucoxanthin ratios are compared with data reported in the literature.
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    Journal of comparative physiology 162 (1992), S. 263-266 
    ISSN: 1432-136X
    Keywords: Metabolism ; Purine nucleotides ; De novo synthesis ; HPLC ; Crustacean, Artemia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In vivo studies of the incoporation of [U-14C]glycine into purine nucleotides have established the de novo pathway for purine biosynthesis in Artemia sp. during the early period of larval development. This pathway can be modified by the salt concentration of the incubation media. In addition, Artemia of different geographical origins may differ with respect to the detection, functionality and variability of this metabolical pathway.
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  • 84
    ISSN: 0884-3996
    Keywords: Luminometers ; radiometers ; low-light level imaging ; review ; survey ; immunoassay ; rapid microbiology ; HPLC ; GLC ; microtitre plate ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: This survey was compiled in January and February 1992 from information available in public domain literature requested by and supplied to the author by numerous companies in the previous two months. More than 90 luminometers (manual, automatic, microtitre plate, HPLC, LC, GLC, imaging and specials) from more than 60 companies are included. Each company was invited to supply company brochures, technical details, user manual and information about software and any other information concerning their product(s). The response varied from a single information sheet to promotional material and up to full product information and specification with technical details, user manuals and scientific publications. Where an instrument is dedicated to a single task the company may have only provided details relevant to accomplishing that task. Part 2 of this survey will contain photographs of some of the luminometers. It is intended that updates to this review will be published at least annually in this journal and suppliers are invited to provide full technical details of new luminometric equipment to the author.
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  • 85
    ISSN: 0884-3996
    Keywords: Luminometers ; radiometers ; low-light level imaging ; review ; survey ; immunoassay ; rapid microbiology ; HPLC ; GLC ; microtitre plate ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Part 1 of this survey comprised a description of the luminometers (J. Biolumin. Chemilumin. 7, 77-108 (1992)) and was compiled in January and February 1992 from information available in public domain literature requested by and supplied to the author by numerous companies in the previous two months. This part includes photographs of around 40 of these instruments. It is intended that updates to this review will be published at least annually in this journal and suppliers are invited to provide full technical details of new luminometric equipment to the author.
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  • 86
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    Archives of Insect Biochemistry and Physiology 21 (1992), S. 77-89 
    ISSN: 0739-4462
    Keywords: HPLC ; protein purification ; embryos ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: From a medium in which Daudi cells had been grown, we isolated by HPLC a protein that caused ovarian abnormalities in adult females of Drosophila melanogaster when injected into preblastoderm embryos. This protein, whose apparent Mr is between 30,000 and 50,000, was found to be a moderately polar compound which is heat stable and whose activity is destroyed by acidification. The protein is characteristic of medium conditioned from Daudi cells. © 1992 Wiley-Liss, Inc.
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  • 87
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    Archives of Insect Biochemistry and Physiology 20 (1992), S. 253-263 
    ISSN: 0739-4462
    Keywords: neuropeptide inhibitors ; antennal heart nerve ; immunocytochemistry ; HPLC ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The presence of allatostatins in the nerves of the antennal pulsatile organ muscle of the cockroach Diploptera punctata was confirmed by immunocytochemistry, bioassay, and HPLC. Immunocytochemical reactivity with monoclonal antibody against allatostatin I showed strong allatostatin immunoreactivity in the antennal heart nerve which innervates this muscle with varicostities along the muscle fibers and in the insertion of the muscle on the pulsatile ampullae. Bioassay of Sep-Pak purified muscle extract demonstrated inhibition of juvenile hormone synthesis by corpora allata in vitro. A dose-response curve showed maximum inhibition of juvenile hormone synthesis was achieved with 10-20 pulsatile organ muscle eq/corpora allata, and 50% inhibition achieved with an estimated 2.6 pulsatile organ muscle eq. Two successive HPLC separations of the Sep-Pak purified extract yielded bioactive fractions corresponding to the elution times of the five known allatostatins. © 1992 Wiley-Liss, Inc.
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  • 88
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    Journal of comparative physiology 169 (1991), S. 39-50 
    ISSN: 1432-1351
    Keywords: Photoreception ; Retinally degenerate ; Mouse ; Circadian ; Rods ; Cones ; 11-cis retinaldehyde ; Immunocytochemistry ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary We have examined the effects of light on circadian locomotor rhythms in retinally degenerate mice (C57BL/6J mice homozygous for the rd allele: rd/rd). The sensitivity of circadian photoreception in these mice was determined by varying the irradiance of a 15 min light pulse (515 nm) given at circadian time 16 and meauring the magnitude of the phase shift of the locomotor rhythm. Experiments were performed on animals 80 days of age. Despite the loss of visual photoreceptors in the rd/rd retina, animals showed circadian responses to light that were indistinguishable from mice with normal retinas (rd/+ and +/+). While no photoreceptor outersegments were identified in the retina of rd/rd animals (80–100 days of age), we did identify a small number of perikarya that were immunoreactive for cone opsins, and even fewer cells that contained rod opsin. Using HPLC, we demonstrated the presence and photoisomerization of the rhodopsin chromophore 11-cis retinaldehyde. The rd/rd retinas contained about 2% of 11-cis retinaldehyde found in +/+ retinas. We have yet to determine whether the opsin immunoreactive perikarya or some other unidentified cell type mediate circadian light detection in the rd/rd retina.
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  • 89
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    Journal of chemical ecology 17 (1991), S. 343-352 
    ISSN: 1573-1561
    Keywords: Allelopathy ; cogongrass ; competition ; Imperata cylindrica ; HPLC ; interference ; weed
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract To understand the interference mechanism of the weed, cogongrass,Imperata cylindrica (L.) Beauv., its effect on nutrient availability and mycoflora of its soil rhizosphere as well as nodule characteristics, root length, and root/shoot ratio of Melilotus parviflora Desf. were investigated. Additionally, the effect of the leachates of leaves and root/rhizome of cogongrass on seed germination and seedling characteristics of radish, mustard, fenugreek, and tomato were examined. Furthermore, to assess the qualitative and quantitative differences in phytochemical components, the leachates and the soils from three sampling sites (with cogongrass and 1.5 m and 3 m away from cogongrass) were analyzed with high-performance liquid chromatography (HPLC) on a C18 column. No significant difference in nutrient availability was found, but qualitative and quantitative differences in phenolic fractions were recorded in the three sampling sites. Furthermore, of the 19 fungi recorded in the soils, decreases in the number of colonies (per gram of soil) ofAspergillus fumigatus, A. niger, A. candidus, and an increase of A. flavus was recorded in the soils with cogongrass. The inhibition in nodule number, weight, nitrogen fixation (acetylene reduction activity), root length, and root/shoot ratio of Melilotus parviflora were noted. Percent seed germination, root and shoot length, fresh and dry weight of seedlings of different seeds were affected by the leachates of leaves and root/rhizome. It was found that root/rhizome leachate was more inhibitory than leaf leachate. However, the inhibition was higher in soil+leaves leachate than soil+root/rhizome leachate. HPLC analysis established that four compounds were contributed by the weed to the soil system even though their relative concentration varies in various leachates. It is surmised that these compounds cause allelopathic inhibition of growth characteristics of seeds tested. Significance of the data vis-a-vis the interference potential of the cogongrass is discussed.
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  • 90
    ISSN: 1432-0878
    Keywords: Brain, vertebrate ; Phenylethanolamine-N-methyltransferase ; Adrenaline ; Immunocytochemistry ; HPLC ; Rat (Wistar, Sprague-Dawley, Long Evans, Zucker)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemistry was used to compare the immunoreactivity of adrenergic neurons to a well characterized specific immunoserum to phenylethanolamine-N-methyltransferase (PNMT) in different strains of rats commonly used in research studies. In adult animals, marked differences were found in the PNMT-immunoreactivity of neurons between Wistar rats and other strains, resulting in a lower PNMT-immunostaining intensity (i) within neuronal perikarya of the medulla oblongata, and (ii) more strikingly, within nerve fibers and terminals located in various brain regions. This low PNMT-immunoreactivity of nerve fibers was detected both in 14- and 35-day-old Wistar rats. On the other hand, the HPLC measurement of catecholamines, in particular of adrenaline in the hypothalamus and the medulla oblongata, did not show any difference between adult Wistar and Sprague-Dawley rats. These data suggest that the low PNMT-immunoreactivity observed in central adrenergic neurons of the Wistar rats is related to the poor recognition of the antigen by the PNMT-antibody used. Possibly, these nerve cells mainly display an isoform of the enzyme that is immunologically different from the PNMT contained within the adrenergic neurons of other rat strains.
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  • 91
    ISSN: 1432-0878
    Keywords: Neurosecretion ; Catecholamines ; HPLC ; Immunohistochemistry ; Glyoxylic acid fluorescence ; Ophryotrocha puerilis (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the posterior part of the brain of the protandric polychaete Ophryotrocha puerilis neurosecretory cells form prominent axon terminals. The terminals are arranged in two complexes. The perikarya of these presumably monopolar neurons are scattered in the anterior part of the cerebral perikaryal layer. In females the terminals store large amounts of neurosecretory material. It has been suggested earlier that neurosecretions of the terminals may play a role during sex reversal from females to males. Application of histamine caused the release of neurosecretory material from the respective terminals in females. However, this discharge was not followed by sex reversal. Application of reserpine had no influence on the terminals. Neither by in vivo observation nor by ultrastructural analysis any effect of reserpine on the terminal complexes could be observed. In isolated terminals filled with neurosecretory material from females, catecholamines could not be detected by HPLC. Also, polyclonal antibodies against dopamine did not stain the terminal complexes. Furthermore, the complexes did not develop any fluorescence after glyoxylic acid treatment. Therefore, the present results contradict the hypothesis that the neurosecretory material of the respective axon terminals is catecholaminergic and that it is involved in sex differentiation. The function of the secretory neurons studied here remains unclear.
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  • 92
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    Journal of applied phycology 3 (1991), S. 259-264 
    ISSN: 1573-5176
    Keywords: vitamin analysis ; HPLC ; algae ; Tetraselmis suecica ; Isochrysis galbana ; Pavlova lutheri ; Skeletonema costatum ; Chaetoceros calcitrans ; Sargassum muticum ; cosmetology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Vitamin analysis was carried out on five microalgae used in aquaculture:Tetraselmis suecica, Isochrysis galbana, Pavlova lutheri, Skeletonema costatum andChaetoceros calcitrans and one macroalga,Sargassum muticum, which is invasive on the Atlantic shores of France. Both liposoluble (provitamin A, E, K) and hydrosoluble (B1, B2, B6, B12, C, PP) vitamins were quantified. For most of them, greater amounts were obtained in the algal products than in the usual sources. On a dry weight basis,Tetraselmis suecica contained 4280 μg g−1 provitamin A and 6323 μg g−1 vitamin E,Pavlova lutheri 1162 μg g−1 vitamin B12 and 837 μg g−1 vitamin C,Isochrysis galbana 2690 μg g−1 vitamin PP and 183 μg g−1 vitamin B6, andSkeletonema costatum 710 μg g−1 vitamin B1.
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  • 93
    ISSN: 1573-5176
    Keywords: determination ; glutamate ; HPLC ; salt interference ; seaweed
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A study was made to find a better method of analyzing the glutamate pool in seaweeds than the use of HPLC, which provides unsatisfactory results with material rich in alginates and salts. A method recommended elsewhere (Inglis A, Bartone N, Finlayson J, 1988, J. Biochem. Biophys. Methods 15: 249–254) for physiological fluids has been assayed and improved for algal samples. It consisted of the addition of lithium acetate before the phenylisothiocyanate derivatization, omission of one drying step and extraction of the derivative with heptane before chromatographic analysis. Neither salt nor alginates interfered with analysis.
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  • 94
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    Hydrobiologia 216-217 (1991), S. 369-376 
    ISSN: 1573-5117
    Keywords: green hydra ; Chlorella ; symbiosis ; amino acids ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Supply of amino acids may be important in controlling cell division of Chlorella symbiotic with green hydra. Freshly isolated symbionts display characteristics of N-limited algae, and low pH in perialgal vacuoles and high levels of host glutamine synthetase (GS) limit uptake of ammonium. Movement of tritiated amino acids from host to algal pools suggests that symbiotic algae utilize amino acids derived from host digestion of prey. Amounts are significant in relation to host and algal amino acids pools. During host starvation, glutamine produced by host GS may be important as a nitrogen supply to the algae, which take up this amino acid at high rates at low pH.
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  • 95
    ISSN: 1573-1561
    Keywords: Wheat ; Triticum aestivum ; soybean ; Glycine max ; no till ; conventional till ; soil extracts ; allelopathy ; phenolic acids ; Folin & Ciocalteu's phenol reagent ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Soil core (0–2.5 and/or 0–10 cm) samples were taken from wheat no till, wheat-conventional till, and fallow-conventional till soybean cropping systems from July to October of 1989 and extracted with water in an autoclave. The soil extracts were analyzed for seven common phenolic acids (p-coumaric, vanillic,p-hydroxybenzoic, syringic, caffeic, ferulic, and sinapic; in order of importance) by high-performance liquid chromatography. The highest concentration observed was 4 μg/g soil forp-coumaric acid. Folin & Ciocalteu's phenol reagent was used to determine total phenolic acid content. Total phenolic acid content of 0- to 2.5-cm core samples was approximately 34% higher than that of the 0- to 10-cm core samples. Phenolic acid content of 0- to 2.5-cm core samples from wheat-no till systems was significantly higher than those from all other cropping systems. Individual phenolic acids and total phenolic acid content of soils were highly correlated. The last two observations were confirmed by principal component analysis. The concentrations were confirmed by principal component analysis, tions of individual phenolic acids extracted from soil samples were related to soil pH, water content of soil samples, total soil carbon, and total soil nitrogen. Indirect evidence suggested that phenolic acids recovered by the water-autoclave procedure used came primarily from bound forms in the soil samples.
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  • 96
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    Biology and fertility of soils 9 (1990), S. 335-340 
    ISSN: 1432-0789
    Keywords: Amino acids ; HPLC ; Immobilized protease ; Organic matter fractions ; Peptides ; Soil nitrogen ; Soil enzymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Organic matter was extracted from three soils, a cultivated Berwick sandy loam, a cultivated Franklin loamy sand, and an uncultivated Cumberland silty loam. Gel-permeation chromatography was used to separate organic matter extracts into high- (HMW) and low-molecular-weight (LMW) fractions. Reversed-phase high performance liquid chromatography was used to separate and collect the LMW peptide fractions. Peptide samples were hydrolyzed with immobilized proteases attached to beaded agarose and carboxymethyl cellulose in column and batch reaction systems. The chromatograms suggested that peptides are bound to common soil components. The amino acids released in the greatest percentages were relatively non-polar. Large percentages of serine, glycine, alanine, threonine, and valine were observed in the LMW soil peptides. Little aspartic acid, asparagine, glutamic acid, glutamine, arginine, and no histidine was detected in the LMW soil peptides. The soil peptides released different amino acid percentages and quantities when hydrolyzed by immobilized proteases attached to different supports. The quanitities of amino acids released by batch hydrolysis differed from those obtained with column hydrolysis. Greater quantities of amino acids were released (by both types of immobilized protease) from the LMW peptide hydrolysates of the two cultivated soils than from the uncultivated soil.
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  • 97
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    Antonie van Leeuwenhoek 57 (1990), S. 179-189 
    ISSN: 1572-9699
    Keywords: heat-resistant moulds ; chemotaxonomy ; HPLC ; TLC ; mycotoxins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Species of the ascomycetous genus Talaromyces have been examined for profiles of secondary metabolites on TLC. The greatest number of specific metabolites were produced on oatmeal-, malt extract- and yeast-extract sucrose agars. Profiles of intracellular secondary metabolites produced on oatmeal agar were specific for each species and provided a means of simple differentiation of the taxa. Examination of the most important species using high performence liquid chromatography (HPLC) allowed to solve some taxonomic problems. Known mycotoxins are produced by T. stipitatus (duclauxin, talaromycins, botryodiploidin), T. stipitatus chemotype II (emodin), T. panasenkoi (spiculisporic acid), T. trachyspermus (spiculisporic acid), T. trac macrosporus (duclauxin) and T. wortmannii (rugulosin). Wortmannin is produced by an atypical strain of T. flavus but not T. wortmannii. Several other secondary metabolites were discovered for the first time in the following species: Glauconic acid is produced by T. panasenkoi, T. ohiensis and T. trachyspermus; vermiculine by T. ohiensis; duclauxin by T. flavus var. macrosporus and the mitorubrins by T. flavus and T. udagawae. The profiles of secondary metabolites support the established taxonomy of the species based on morphology, showing the genetic stability of profiles of secondary metabolites in Talaromyces. Two new taxa are proposed: T. macrosporus comb. nov. (stat. anam. Penicillium macrosporum stat. nov.), and Penicillium vonarxii, sp. nov. for the anamorph of T. luteus.
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    Archives of microbiology 153 (1990), S. 432-437 
    ISSN: 1432-072X
    Keywords: Chromatium vinosum ; Phototrophic bacteria ; Polysulfides ; Polythionates ; Elemental sulfur ; Sulfur globules ; Ion chromatography ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cultures of Chromatium vinosum, devoid of sulfur globules, were supplemented with sulfide and incubated under anoxic conditions in the light. The concentrations of sulfide, polysulfides, thiosulfate, polythionates and elemental sulfur (sulfur rings) were monitored for 3 days by ion-chromatography and reversed-phase HPLC. While sulfide disappeared rapidly, thiosulfate and elemental sulfur (S6, S7 S8 rings) were formed. After sulfide depletion, the concentration of thiosulfate decreased fairly rapidly, but elemental sulfur was oxidized very slowly to sulfate. Neither polysulfides (S x 2− ), polythionates (SnO 6 2− , n=4–6), nor other polysulfur compounds could be detected, which is in accordance with the fact that sulfide-grown cells were able to oxidize polysulfide without lag. The nature of the intracellular sulfur globules is discussed.
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  • 99
    ISSN: 1573-1561
    Keywords: Asclepias fruticosa ; milkweed ; Danaus plexippus ; monarch butterfly ; Lepidoptera ; Danaidae ; cardenolides ; HPLC ; gomphoside ; afroside ; digitoxin ; calactin ; calotropin ; cardenolide fingerprint ; cardiac glycosides ; internal standard
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The cardenolide extracts from latex and aerial parts ofAsclepias fruticosa and ofDanaus plexippus reared onA. fruticosa orA. curassavica were purified by adsorption chromatography on silica gel. HPLC analysis on a C18 reverse-phase column with an acetonitrile-water gradient as mobile phase, separated 28 compounds with a UV spectrum typical forcardenolides. Afroside and gomphoside (major components), as well as calotropagenin, calotoxin, calotropin, calactin, uscharidin, uscharin, and voruscharin, occurred as single peaks in the profiles of latex and aerial plant parts ofA. fruticosa. Calactin and calotropin were the major cardenolides inDanaus plexippus reared onA. fruticosa orA. curassavica. Quantitative data obtained with digitoxin as internal standard showed that 1.3–1.5% of the leaf cardenolides were sequestered byDanaus plexippus in which levels of 70–80μg cardenolide per butterfly were measured. The calotropin from the leaves was almost completely sequestered, and 10–13% of the calactin was stored by the butterfly, assuming that no conversion occurred in larval tissues.
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  • 100
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    Protoplasma 156 (1990), S. 130-138 
    ISSN: 1615-6102
    Keywords: Glyoxysomes ; Castor bean ; Enzyme purification ; Catalase ; HPLC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A strategy for the rapid purification of proteins from glyoxysomes of castor bean (Ricinus communis cv. Hale) is described. The first step was to separate the proteins in the mixture on the basis of hydrophobicity by reversed phase high performance liquid chromatography using a gradient of increasing acetonitrile concentration. Individual protein peaks were collected and fractionated according to molecular mass by preparative polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The purified polypeptides were used to produce monospecific, polyclonal antibodies. One of these, an anti-catalase antibody, has been employed to assess the subcellular distribution of catalase in endosperm of maturing seeds, dry seeds and seedlings. During seed maturation 45% of the catalase activity was associated with structures sedimenting at high isopycnic densities (1.21 g/cm3). However, in dry seeds, only 6% or less of the catalase activity was associated with these dense particles. In 4-day seedlings 80% of catalase activity was associated with glyoxysomes (1.24 g/cm3). A novel catalase 59 kDa subunit was found in the cytosol of 4-day seedlings and in isolated organelles from maturing and dry seed.
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