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  • Articles  (591)
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  • Calcification
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 2 (1968), S. 296-298 
    ISSN: 1432-0827
    Keywords: Calcium ; Lipid ; Bacteria ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé On a fait ce travail pour determiner si le facteur responsable pour la liaison de calcium par un calcifableBacterionema matruchotii est dans la fraction lipide de la cellule. Des cellules congelees et sechees ont ete extraites par le chloroform-methanol. La fraction de chloroform-methanol, les cellules extraites et les cellules non traitees ont ete examinees pour la liaison de calcium. La fraction du chloroform-methanol et les cellules non traitees avaient la liaison de calcium. Les cellules extraites n'en avaient pas.
    Abstract: Zusammenfassung Diese Arbeit wurde durchgeführt um festzustellen, ob sich der Faktor für die Calcium-bindung, durch das calcifizierendeBacterionema matruchotii, in der Lipoidfraktion befindet. Die lyophiilisierten Zellen wurden mit Chloroform-Methanol extrahiert. Die Chloroform-Methanol-Fraktion, die extrahierten Zellen, sowie die nicht behandelten Zellen wurden auf eine Calciumbindung hin untersucht. Die Chloroform-Methanol-Fraktion und die nicht behandelten Zellen demonstrierten eine Calciumbindung. Die extrahierten Zellen hingegen nicht.
    Notes: Abstract This work was done to determine whether the factor responsible for calcium binding by a calcifiableBacterionema matruchotii is in the lipid fraction of the cell. Freeze-dried cells were extracted with chloroform-methanol. The chloroform-methanol fraction, the extracted cells and untreated cells were examined for calcium binding. The chloroform-methanol fraction and the untreated cells bound calcium. The extracted cells did not.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 4 (1969), S. 260-268 
    ISSN: 1432-0827
    Keywords: Cartilage ; Histochemistry ; Staining ; Protein ; Polysaccharide ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des coupes de cartilage épiphysaire frais de jeunes rats, effectuées à la main, sont colorées à pH=4,5 dans des solutions à 0,01% de divers colorants cationiques, appartenant aux groupes de la thiazine, oxazine, azine, triphénylméthane, acridine, et phthallocyanine. Les granules intracellulaires métachromatiques, mises en évidence antérieurement par le bleu de toluidine, sont également identifiées à l'aide de l'azur A, le bleu de méthylène et le bleu de crésyl. Les granules se colorent moins bien à la thionine, le rouge neutre, la safranine O, le bleu de toluylène et l'acridine orange. Dans les conditions utilisées, la matrice de la zone de réserve et la matrice de la zone hypertrophique inférieure (en voie de calcification) se colorent, alors que les matrices des zones prolifératives et hypertrophiques supérieures ne prennent pas les colorants. La gallocyanine, le violet cristal, la fuchsine basique, l'azocarmin B, le bleu de gallamine et la bleu alcian ne se colorent pas ou donnent des réactions colorées différentes de celles décrites ci-dessus. Il semble que le pK et le poids moléculaire des colorants jouent un rôle important, mais ils ne paraissent pas être les seuls facteurs intervenant dans la coloration des granules. Un changement, lié à la calcification, semble intervenir au niveau du matériel métachromatique (probablement des polysaccharides protéiques), aussi bien dans la matrice que les cellules cartilagineuses épiphysaires.
    Abstract: Zusammenfassung Handpräparierte Schnitte von frischem Epiphysenknorpel junger Ratten wurden bei einem pH von 4,5 in 0,01% igen Lösungen verschiedener kationischer Farbstoffe folgender Klassen gefärbt: Thiazin, Oxazin, Azin, Triphenylmethan, Acridin und Phthalocyanin. Die intracellulären β-und γ-metachromatischen Granula, erstmals mit Toluidinblau im frischen Gewebe nachgewiesen, konnten auch gut mit Azur A, Methylenblau und Brillantkresylblau dargestellt werden. Die Granula konnten ebenfalls, aber weniger gut, mit Thionin, Neutralrot, Safranin D, Toluylenblau und Acridinorange gefärbt werden. Unter diesen Färbungsbedingungen werden die inaktive Matrixzone und die untere hypertrophische (verkalkende) Matrixzone angefärbt, während die proliferative und die obere hypertrophische Matrixzone sich nicht färben. Gallocyanin, Kristallviolett, basisches Fuchsin, Azokarmin B, Gallaminblau und Alzianblau färbten entweder gar nicht, oder gaben ein anderes als das obenbeschriebene Färbemuster. Es wird vorgeschlagen, daß das pK und das Molekulargewicht der Farbstoffe wichtig aber nicht unbedingt die einzigen Faktoren sind, die die Färbung der Granula bestimmen. Die Resultate zeigen, daß eine Veränderung im metachromatischen Material (vermutlich Proteinpolysaccharide) vorliegt, und zwar sowohl in der Matrix als in den Zellen des Epiphysenknorpels; diese Veränderung scheint im Zusammenhang mit der Verkalkung zu stehen.
    Notes: Abstract Hand-cut sections of fresh epiphyseal cartilage from young rats were stained at pH 4.5 in 0.01% solutions of various cationic dyes of the thiazine, oxazine, azine, triphenylmethane, acridine, and phthallocyanin classes. The intracellular β-and γ-metachromatic granules, previously demonstrated in fresh tissues with toluidine blue, were also demonstrated well with azure A, methylene blue, and brilliant cresyl blue. The granules were also demonstrated, but not as well, by thionin, neutral red, safranin O, toluylene blue, and acridine orange. Under the conditions of staining, the reserve zone matrix and the lower hypertrophic (calcifying) zone matrix stained, whereas the proliferative and upper hypertrophic zone matrix did not stain. Gallocyanin, crystal violet, basic fuchsin, azocarmine B, gallamine blue, and alcian blue either did not stain, or gave a different pattern of staining from that described above. It is suggested that the pK and molecular weight of the dyes are important, but not necessarily the only factors in determining the staining of the granules. The results indicate that there is a change in the metachromatic material (presumably proteinpolysaccharide) in both the matrix and cells of epiphyseal cartilage, which appears to be related to calcification.
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  • 3
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    Springer
    Calcified tissue international 4 (1969), S. 20-38 
    ISSN: 1432-0827
    Keywords: Calcification ; Epiphyseal Cartilage ; Bone ; Electrolytes ; Organic matrices
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Un procédé de dissection a été mis au point pour permettre l'analyse zonale du cartilage de l'épiphyse des os de la jambe d'un foetus bovin. Des échantillons de tissu complet et lavé venant des différentes zones ont été analysés pour déterminer leur contenu en électrolyte et en constituants organiques, ainsi que pour leur densité, cendres et humidité. Les résultats ont montré que lorsque la quantité de cendres et la densité augmentaient, l'eau contenu dans le tissu diminuait. Les quantités de cendres dans les zones de cartilage en voie de calcification étaient plus grandes qu'il avait été. Quand elles étaient exprimées comme un pourcentage du poids sec, elles étaient les plus importantes dans le cartilage lavé calcifié que dans le autre zones. Au début de la minéralisation du cartilage, la quantité de Na (m moles/l de tissu frais) diminuait tandis que celles du Ca et du P inorganique augmentaient. Les niveaux de Mg augmentaient pendant que la calcification se poursuivait, mais seulement à une faction du taux du Ca et du P. Les rapports Ca/P inorganique étaient les plus grands dans le cartilage au repos (Cartilage non-différentié hyalin), suggérant un lien initiale entre Ca et les chrondromucoprotéines. Cependant, au début de la calcification, pendant la prolifération du cartilage les rapports Ca/P étaient beaucoup plus petits (ca. 1.50) mais augmentaient graduellement avec l'advancement de la minéralisation. Des changements importants survenaient dans la composition de la phase organique, pendant la calcification endochondrale. Comme il a été déterminé par l'analyse de l'hydroxyproline la quantité de collagéne diminuait progressivement pendant la calcification du cartilage, mais augmentait rapidement pendant la formation d'os. Comme il a été déterminé par l'analyse de l'héxosamine et du sulfute les chrondromucoprotéines étaient aux niveaux les plus éléves pendant la prolifération du cartilage et diminuaient constamment au cours de la calcification. Cependant, bien que la calcification était déja très avancée dans le cartilage hypertrophique, de grandes quantites de mucopolysaccharides étaient encore présentes. Les rapports sulfure/hhéxosamine montraient un léger déclin pendant les premiéres étapes de la calcification, mais augmentaient beaucoup pendant le cours de la minéralisation. Les quantités d'acide sialique étaient plus grandes dans le cartilage de l'épiphyse que dans le cartilage au repos ou dans l'os. Les lipides augmentaient rapidement pendant la calcification du cartilage, mais étaient très réduites dans l'os complètement formé. La signification de ces résultats est discutée.
    Abstract: Zusammenfassung Eine Seziermethode, die eine Schichten-Analyse der Beinepiphysenplatte von Rinderfeten erlaubt, wurde entwickelt. Proben vor und nach Waschen des Gewebes der verschiedenen Schichten werden untersucht in bezug auf Elektrolyte und organische Bestandteile, als auch in bezug auf Dichte, Aschengehalt und Feuchtigkeit. Die Resultate zeigten eine Zunahme des Aschengehaltes und der Dichte, während der Wassergehalt abnahm. Unerwartet hoch waren die Aschenwerte im in Verkalkung begriffenen Knorpel. Ausgedrückt in Prozent Trockengewicht, ergab gewaschener, verkalkter Knorpel den höchsten Wert aller Zonen. In den Frühstadien der Knorpelmineralisation nahm der Natriumgehalt (m Mol/l Frischgewebe) ab, während Ca und anorganischer P zunahmen. Mit fortschreitender Verkalkung erhöhte sich auch der Magnesium-Spiegel, allerdings nur zu einem Bruchteil des Ausmaßes, in welchem Ca und P zunahmen. Die höchsten Ca/P anorg. Verhältnisse wurden im Ruheknorpel (undifferenzierter hyaliner Knorpel) gefunden, was auf eine initiale Bindung von Ca durch Chondromucoproteine hinweist. Die Ca/P-Verhältnisse proliferierenden Knorpels waren jedoch bei Verkalkungsbeginn viel tiefer (ca. 1.50). Diese nahmen allerdings mit fortschreitender Mineralisierung stetig zu. In der endochondralen Verkalkungsphase fanden markante Veränderungen in der Zusammensetzung des organischen Anteils statt. Basierend auf der Hydroxyprolinanalyse nahm der Collagengehalt in der knorpeligen Verkalkungsperiode fortschreitend ab, während er jedoch bei der Knochenbildung rasch zunahm. Die an Hand von Hexosamin- und Schwefelanalysen bestimmten Chondromucoproteingehalte ergaben Höchstwerte im proliferierenden Knorpel und fielen stetig ab mit zunehmender Verkalkung. Trotz der im hypertrophischen Knorpel schon weit fortgeschrittenen Verkalkung waren immer noch große Mengen an Mucopolysacchariden vorhanden. Die Schwefel/Hexosamin-Verhältnisse zeigten eine minimale Abnahme in den frühen Verkalkungsphasen, nahmen jedoch markant zu bei fortschreitender Mineralisation. Der Sialinsäurespiegel war im Epiphysenknorpel, verglichen mit demjenigen des Ruheknorpels oder Knochens, erhöht. In der knorpeligen Verkalkungsphase nahmen die Lipide rasch zu, während jedoch die Werte des vollständig ausgebildeten Knochens stark vermindert waren. Die Bedeutung dieser Ergebnisse wird besprochen.
    Notes: Abstract A dissection procedure has been devised to permit zonal analysis of the epiphyseal plate of fetal calf leg bones. Samples of whole and washed tissue from the various zones were analyzed for their content of electrolyte and organic constituents, as well as for density, ash and moisture. Results showed that as ash content and density increased, water content decreased. Ash levels in calcifying cartilage zones were unexpectedly high. When expressed as a percentage of dry weight, washed calcified cartilage had the highest content of any zone. In the early stages of the mineralization of cartilage, Na content (mmoles/l of fresh tissue) decreased as Ca and inorganic P increased. Magnesium levels increased as calcification proceeded, but only at a fraction of the rate of Ca and P. Ratios of Ca/inorganic P were highest in resting cartilage (non-differentiated hyaline cartilage), suggesting an initial binding of Ca to chondromucoproteins. However, at the onset of calcification in proliferating cartilage, Ca/P ratios were much lower (ca. 1.50), but gradually increased with advancing mineralization. Marked changes occurred in the composition of the organic phase during endochondral calcification. As determined by hydroxyproline analysis, collagen content progressively decreased during cartilaginous calcification, but increased rapidly during bone formation. As determined by hexosamine and sulfur analysis, chondromucoproteins were at highest levels in proliferating cartilage and decreased steadily as calcification increased. However, although calcification was already well advanced in hypertrophic cartilage, large amounts of mucopolysaccharide still were present. Sulfur/hexosamine ratios showed a slight decline during the early stages of calcification, but increased markedly with further mineralization. Sialic acid levels were elevated in epiphyseal cartilage over those in resting cartilage or bone. Lipids increased rapidly during cartilaginous calcification, but were greatly reduced in fully-formed bone. The significance of these findings is discussed.
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  • 4
    ISSN: 1432-0975
    Keywords: Key words Coral ; Calcification ; Biomineralisation ; Autoradiography ; Freeze-substitution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract  The uptake of 45Ca and/or 14C by the skeleton of coral colonies has been commonly used to investigate the processes of calcification. This study reports the differential uptake of these tracers within different regions of the skeleton and tissues of individual corallites and polyps of the hermatypic coral Galaxea fascicularis. Incubation in 45Ca in the light resulted in 80 percent of the 45Ca taken up being deposited in the skeleton. Autoradiography of transverse and longitudinal slices of freeze-substituted polyps and corallites showed that in the light 45Ca was incorporated into the exsert septa, the outside of the thecal walls of the corallite and the inner edges of the septa. Incorporation did not occur in the costae. The radioactivity in the skeleton was considerably greater than in the tissues. In the dark, or in the presence of the photosynthetic inhibitor Diuron, 45Ca was taken up by the exsert septa and was patchily distributed in the corallite walls which suggests that it was not a result of isotopic exchange. The differential incorporation of 45Ca onto the exsert septa was confirmed by scintillation counting. Negligible radioactivity remained in the extrathecal coelenteron after a brief 5 min rinse in non-radioactive seawater. Only 0.1% of 14C taken up in the light was incorporated into the skeleton and this was confirmed by autoradiography. In the presence of Diuron or in the dark, very little 14C was incorporated into tissues or skeleton and in autoradiographs was either not evident in the skeleton or the distribution was similar to that seen in autoradiographs of 45Ca uptake. These results show that the deposition of 45Ca, and therefore calcium carbonate, occurs at specific loci on the skeleton of a corallite. In the dark, deposition occurs specifically at the growing points of the corallite. Differential deposition of calcium carbonate within individual corallites has not been previously reported.
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  • 5
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    Mangroves and salt marshes 2 (1998), S. 99-107 
    ISSN: 1572-977X
    Keywords: conductance ; mangrove ; photosynthesis ; productivity ; water potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Diurnal gas exchange characteristics were measured simultaneously in two mangrove species, Avicennia marina and Bruguiera gymnorrhiza, over 7 d in summer (February–March), to compare their productivity. The study was undertaken in the Beachwood Mangroves Nature Reserve, Durban, South Africa, using fully expanded leaves of young and mature trees at the top of the canopy. Gas exchange was strongly influenced by photosynthetic photon flux density (PPFD), leaf temperature and the accompanying leaf to air vapour pressure deficit (Δ w). Carbon dioxide exchange was saturated at a PPFD of about 600 μmol m-2s-1 in B. gymnorrhiza compared to 800 μmol m-2s-1 in A. marina. Maximal CO2 exchange occurred between 12h00 and 14h00 and was consistently greater in A. marina (8.8 μmol m-2s-1) than in B. gymnorrhiza (5.3 mu;mol m-2s-1). Mean internal CO2 concentrations ( ci) were 260 μl l-1 in A. marina and 252 μl l-1 in B. gymnorrhiza. Photorespiratory activity was 32% in A. marina and 30% in B. gymnorrhiza. Mean water use efficiency (WUE) was 8.0 μmol mmol-1 in A. marina and 10.6 μmol mmol-1 in B. gymnorrhiza. Diurnal leaf water potentials ranged from –0.8 to –3.5 MPa and were generally lower in A. marina.
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  • 6
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    Mangroves and salt marshes 2 (1998), S. 191-198 
    ISSN: 1572-977X
    Keywords: canopy ; Hinchinbrook ; leaf area index ; mangrove ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Data on stand structure and rates of photosynthesis were used to estimate net canopy carbon fixation and carbon accumulation as living biomass in mangrove forests in Hinchinbrook Channel, Australia. Total annual canopy net carbon fixation was estimated to be about 29 t C ha−1 yr−1. This equates to about 204,000 t C yr−1 for all mangrove forests in Hinchinbrook Channel. Of this, only about 12% was stored as living plant biomass. Although it is not yet possible to present a robust carbon balance for mangrove trees, the remainder is presumably lost through plant respiration, litter fall, root turnover and exudation of organic compounds from roots.
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  • 7
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    Mangroves and salt marshes 3 (1999), S. 147-153 
    ISSN: 1572-977X
    Keywords: conductance ; gas exchange ; mangrove ; photorespiration ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photosynthetic characteristics were investigated in the geographically isolated and restricted mangrove species, P.rhizophoreae. Gas exchange measurements were made on two to seven years old hydroponically grown plants maintained in 10%, 50% and 100% seawater. CO2 exchange in the 50% and 100% seawater treatments was reduced by 10% and 26%, respectively, compared to the 10% seawater treatment. CO2 response curves indicated that carboxylation efficiency was greater in 10% than in 50% seawater, while stomatal limitation increased from 11% to 16% as salinity increased from 10% to 50% seawater. Carbon losses via photorespiration (31% and 41%) and CO2 compensation point (67 and 81 μ11−1) were greater in 50% than in the 10% seawater treatment. Maximal CO2 exchange occurred at 30 °C with no differences among the salinity treatments. The results indicate that P. rhizophoreae exhibits many gas exchange characteristics previously reported for other mangroves.
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  • 8
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    Calcified tissue international 3 (1969), S. 151-162 
    ISSN: 1432-0827
    Keywords: Calcification ; Physiologic ; Phosphonic Acids ; Phosphates ; Crystallization ; Electron Microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé On a étudié la formation de l'hydroxyapatite de calcium cristallin à partir de solutions d'ions de calcium et de phosphate et l'inhibition de la croissance de cristaux de l'hydroxyapatite de calcium au moyen de polyphosphonates et de polyphosphates. Les polyphosphonates, éthane hydroxy-1-diphosphonate-1,1 de disodium et dichlorométhanediphosphonate de disodium, sont inhibiteurs efficaces contre la croissance de cristaux de l'hydroxyapatite de calcium. Les polyphosphates sont aussi inhibiteurs efficaces contre la croissance de cristaux de l'hydroxyapatite de calcium tant que le niveau exigé de polyphosphate intact est présent dans le système. Cependant, à cause de leur instabilité hydrolytique, qui est soulignée par une température élevée, valeur de pH basse, et certaines enzymes, la concentration du polyphosphate diminue avec le tempsin vitro, et son activité comme inhibiteur est perdue. Au contraire aux polyphosphates, les polyphosphonates sont hydrolytiquement stables. Les polyphosphonates sont chimiosorbés sur la surface des microcristallites de l'hydroxyapatite de calcium, ainsi empêchant l'occurrence d'autre croissance de cristaux semblable à l'action d'autres poisons connus de croissance de cristaux. On propose l'extension de cette action sur la formation de l'apatite et cette stabilité des polyphosphonates aux applications médicales et dentaires concernant le metabolisme pathologique de calcium et de phosphate.
    Abstract: Zusammenfassung Die Bildung des kristallinen Calciumhydroxyapatit aus Lösungen, welche Calcium- und Phosphationen enthalten, und die Hemmung der Bildung von kristallinen Calciumhydroxyapatit durch Polyphosphonate und Polyphosphate wurden untersucht. Polyphosphonate, Dinatriumäthan-1-hydroxyl-1,1-diphosphonat und Dinatriumdichloromethandiphosphonate verhindern das Kristallwachstum des Calciumhydroxyapatits. Die Polyphosphate verhindern ebenfalls das Kristallwachstum des Calciumhydroxyapatits, solange die notwendige Konzentration des nicht hydrolysierten Polyphosphats vorhanden ist. Wegen ihrer hydrolytischen Unbeständigkeit, die durch hohe Temperatur, niedrige pH und bestimmte Enzyme erhöht wird, vermindert sich jedoch die Konzentration des Polyphosphats allmählichin vitro, und ihre Hemmungsaktivität geht verloren. Im Gegensatz zu den Polyphosphaten sind die Polyphosphonate hydrolytisch beständig. Die Polyphosphonate werden an der Oberfläche der Mikrokristallite des Calciumhydroxyapatits chemisorbiert und verhindern, wie andere bekannte Kristallwachstumsgifte, auf diese Weise weiteres Kristallwachstum. Die Beständigkeit der Polyphosphonate und ihre Chemisorption an dem Apatit empfehlen ihren Gebrauch in der ärztlichen und zahnärztlichen Praxis, soweit sie den pathologischen Calcium- und Phosphatstoffwechsel betreffen.
    Notes: Abstract The formation of crystalline calcium hydroxyapatite from solutions of calcium and phosphate ions and the inhibition of calcium hydroxyapatite crystal growth by polyphosphonates and polyphosphates have been studied. The polyphosphonates, disodium ethane-1-hydroxy-1,1-diphosphonate and disodium dichloromethane diphosphonate, are effective inhibitors of calcium hydroxyapatite crystal growth. The polyphosphates are also effective inhibitors of calcium hydroxyapatite crystal growth as long as the required level of intact polyphosphate is present in the system. However, because of their hydrolytic instability, which is enhanced by high temperature, low pH, and certain enzymes, the concentration of the polyphosphate decreases with timein vitro, and its activity as an inhibitor is lost. In contrast to the polyphosphates, the polyphosphonates are hydrolytically stable. The polyphosphonates are chemisorbed on the surface of the microcrystallites of calcium hydroxyapatite and, in the manner of other known crystal growth poisons, thus prevent further crystal growth. The stability of the polyphosphonates and their chemisorption on apatite suggest their use in medical and dental applications involving pathological calcium and phosphate metabolism.
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  • 9
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    Calcified tissue international 3 (1969), S. 184-193 
    ISSN: 1432-0827
    Keywords: Mitochondria ; Calcification ; Calcium ; Chondrocytes ; Growth plate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des cultures d'épiphyse prélevées sur des rats et des souris ont été étudiées sous le microscope électronique. Un profile de granules mitochondriques de densité électronique a été trouvé. Les chondrocytes dans la zone proliférative avaint peu de granules, alors que ceux des zones successives ont montré une augmentation progressive de leur nombre et densité jusqu'à ce que la zone de calcification provisoire ait été atteinte. Cette zone a montré une distribution périphérique de mitochondries et une réduction du nombre et de la densité des granules mitochondriques. Du calcium isotopique 47 a été utilisé autoradiographiquement pour déterminer la location de calcium dans ces cellules. Des grains ont été trouvés sur les membranes R.E. et sur la plupart des mitochondries. La preuve d'un profile de ces granules et de leur rapport spatial avec la face de minéralisation indique une action éventuelle de mitochondries dès le début de la calcification de la matrice.
    Abstract: Zusammenfassung Epiphysekulturen von Ratten und Mäusen wurden unter dem Elektronenmikroskop untersucht. Ein Profil von mitochondrischen Körnchen mit elektronischer Dichte wurde gefunden. Chondrozyten in der Proliferationszone wiesen wenig Körnchen auf, während die der nachfolgenden Zonen allmählich an Zahl und Dichte zunahmen, bis die Zone der provisorischen Verkalkung erreicht wurde. Diese Zone zeigte eine periphere Verteilung der Mitochondrien und eine Abnahme in Zahl und Dichte der mitochondrischen Körnchen. Isotopes Kalzium 47 wurde autoradiographisch verwendet, um die Lage des Kalziums in diesen Zellen zu bestimmen. Körnchen wurden auf den E.R.-Membranen und auf einem Großteil der Mitochondrien gefunden. Der Nachweis eines Profils dieser Körnchen und ihres räumlichen Verhältnisses zur Mineralisierungsfläche weist auf einen möglich Einfluß der Mitochondrien mit Beginn der Matrixverkalkung hin.
    Notes: Abstract Rat and mice epiphyseal growth plates were studied with the electron microscope. A gradient of mitochondrial electron-dense granules was found. Chondrocytes in the proliferative zone had few granules, while those of the succeeding zones showed a gradual increase in number and density until the zone of provisional calcification was reached. This zone showed a peripheral distribution of mitochondria and a decrease in the number and density of mitochondrial granules. Isotopic47calcium was used autoradiographically to determine the location of calcium in these cells. Grains were found over the endoplasmic reticulum membranes and over most mitochondria. The demonstration of a gradient of these granules and their spatial relation to the mineralization front suggests a possible involvement of mitochondria in the onset of matrix calcification.
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  • 10
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    Calcified tissue international 4 (1969), S. 78-83 
    ISSN: 1432-0827
    Keywords: Bone ; Calcification ; Osteomalacia ; Phosphorus ; Vitamin D
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les effects d'une dose de 1 mg de vitamine D3 (40 000 Unités,) administrée par voie intra-veineuse, ont été étudiés sur l'histologie osseuse et le métabolisme du phosphore chez 19 sujets contrôles dont l'histologie était normale et 28 malades présentant les caractères histologiques de l'ostéomalacie par carence vitaminique D. L'administration de la vitamine D n'a entraîné aucune modification histologique ou biologique significative chez les sujets contrôles. Mais chez les ostéomalaciques, il est apparu en moins de sept jours une augmentation très significative du front de calcification à l'interface tissue-ostéoïde-tissue-calcifié. Cette modification s'accompagnait d'une ascension progressive de la phosphatémie et de la réabsorption tubulaire du phosphore atteignant dans le même délai des valeurs normales.
    Abstract: Zusammenfassung Bei 19 Patienten mit normaler Knochenhistologie und bei 28 Patienten mit histologisch gesicherter Osteomalacie wurde die Wirkung von 1 mg Vitamin D3 (40000 Iv) i.v. auf die Knochenhistologie und den Phosphatmetabolismus untersucht. Bei den Kontrollpatienten konnten keine signifikanten Änderungen nach Vitamin D festgestellt werden, wogegen die Osteomalacie-patienten innerhalb von 7 Tagen eine deutliche Zunahme der Verkalkungszone an der Grenze zwischen Osteoid- und Knochengewebe zeigten. Diese Änderung war von einer fortschreitenden Zunahme des Serum-Phosphates, verbunden mit einer gesteigerten renalen tubulären Rückabsorption des Phosphates begleitet; beide kehrten anschließend gleichzeitig wieder zur Norm zurück.
    Notes: Abstract The effect of a 1 mg dose of intravenous Vitamin D3 (40,000 i.u.) on bone histology and phosphate metabolism was investigated in 19 patients with normal bone histology and 28 patients with histological evidence of osteomalacia. No significant changes occurred in the control patients after Vitamin D but the patients with osteomalacia showed a marked increase, within seven days, in the proportion of osteoid having a calcification front. This was accompanied by a progressive rise in the serum phosphate, which was associated with an increase in the renal tubular reabsorption of phosphate to normal.
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    Calcified tissue international 4 (1969), S. 231-244 
    ISSN: 1432-0827
    Keywords: Calcification ; Pyrophosphate ; Inhibitors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les propriétés d'induction de calcification des liquides biologiques peuvent être mesurées, in vitro et in vivo, par leur possibilité de redurcir des blocs d'émail décalcifié. Cette propriété est exprimée par les concentrations de calcium (et de phosphate) de solutions synthétiques inductrices de calcification, de concentrations et d'activité connues. Un plasma humain typique a une activité inductrice correspondant à celle d'une solution contenant du calcium, 0,70 mM, Ca/P, 1,67 et fluorure, 0,05 mM. L'activité relativement peu élevée des sérums et des plasmas s'explique par la présence d'inhibiteurs ioniques. L'ion phosphate est l'un d'eux, mais ne peut rendre compte de l'inhibition totale. Des anions et des cations sont responsables de l'inhibition, avec un rôle majeur pour les cations. Les ions suivants, en concentration physiologique, jouent un rôle inhibiteur: P7O 7 4− , HCO 3 − , SiO 4 2− , CrO 4 2− , Mg2+, Zn2+. Un mélange de ces ions provoque une inhibition totale identique à celle du plasma.
    Abstract: Zusammenfassung Der Einfluß biologischer Flüssigkeiten auf den Verkalkungsvorgang kann in vitro und in vivo anhand ihrer Fähigkeit, enthärtete Blöcke von Zahnschmelz wieder zu härten, gemessen werden. Diese Aktivität wird ausgedrückt als Konzentration des Calciums (und Phosphates) synthetischer calcifizierender Lösungen mit bekannter Konzentration und Aktivität. Die Aktivität eines charakteristischen menschlichen Plasmas entsprach derjenigen einer Lösung folgender Zusammensetzung: Calcium 0,70 mM; Ca/P=1,67 und Fluorid 0,05 mM. Die relativ niedrige Aktivität von Serum und Plasma ist bedingt durch das Vorhandensein einer Anzahl ionischer Inhibitoren. Das Pyrophosphation ist ein solcher; er kann aber nicht für die gesamte Inhibition verantwortlich gemacht werden. Sowohl anionische als auch kationische Inhibitoren sind vorhanden, wobei die Kationen den Hauptanteil ausmachen. Folgende Ionen erwiesen sich in einer physiologischen Konzentration als Inhibitoren synthetischer Systeme: P7O 7 3− , HCO 3 − , SiO 4 1− , CrO 4 2− , Mg2+, Zn2+. Zusammengenommen verursachten diese Ionen eine Gesamtinhibition ähnlich derjenigen des Plasmas.
    Notes: Abstract The calcifying activities of biological fluids can be measured,in vitro andin vivo, by their ability to reharden softened blocks of tooth enamel. The activity is expressed in terms of the calcium (and phosphate) concentrations of synthetic calcifying solutions of known concentration and activity. A typical human plasma had an activity corresponding to that of a solution of the following concentration: calcium, 0.70 mM; Ca/P, 1.67 and fluoride, 0.05 mM. The relatively low activity of serums and plasmas was shown to arise from the presence of a number of ionic inhibitors. Pyrophosphate ion is one such inhibitor but cannot account for the major inhibition. Both anionic and cationic inhibitors were shown to be present, with the cations respresenting the major portion. The following ions at their reported physiological concentration were shown to be inhibitors in synthetic systems: P7O 7 4− , HCO 3 − , SiO 4 2− , CrO 4 2− , Mg2+, Zn2+. In combination, these ions caused a total inhibition similar to those for plasmas.
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    Calcified tissue international 3 (1969), S. 266-273 
    ISSN: 1432-0827
    Keywords: Calcification ; Arteries ; Membranes ; Elastic tissue
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Avec la modification d'une méthode de von Kossa nous avons macroscopiquement representé les dessins de calcification de la membrane élastique intérieure, ainsi que les grosses artères du bassin, des artères des extrémités inferieures, des artères du bras, de la rate et des reins. Dans les artères du type élastique, dans les A.a. ilicae communes et internae, se présentaient les dégénérations calcaires de la membrane élastique intérieure sous forme d'incrustations noires arrondies ou polygonales qui ont été formes en groupes. Elles étaient toujours présentes chez des enfants agés de plus de trois mois et chez des aux adultes. Dans les artères musculaires, se montraient les parties calcifiées de la membrane élastique intérieure en forme de «bandes de calcaire» noires et groupes en paires le long des bordures des fissures de la membrane. Dans le cas où existait une calcinose forcée, on pouvait montrer avec cette méthode tout le système de fissures de la mambrane élastique intérieure. Chez tous les morts âgés de 10 à 20 ans, nous avons trouvé des bandes de calcaire dans les artères musculaires des extrémités inferieures. Ces bandes de calcaire ont été constatées aussi dans des personnes plus âgées. Au point de la fusion des bandes de calcaire et des incrustations calcaire polygonales des dépôts calcaires en forme de feuille («feuilles de calcaire») se forment dans la membrane elastique. Les bandes de calcaire et les feuilles de calcaire représentent des points de cristallisation pour des dépôts calcaire granuleux. Ils se forment dans la plupart des cas sur le la côté externe tourné vers la media.
    Abstract: Zusammenfassung Mit einer modifizierten von Kossa-Methode wurden die Verkalkungsmuster der inneren elastischen Membran der großen Beckenarterien, der Arterien der unteren Extremität, der Oberarm-, Milz-und Nierenarterien makroskopisch dargestellt. In den Arterien vom elastischen Typ, i.e. in den Aa. ilicae communes et internae, erschienen die Verkalkungen der inneren elastischen Membran als schwarze rundliche oder polygonale Inkrustationen, die zu Gruppen angeordnet waren. Sie wurden häufig bereits bei Neugeborenen festgestellt. Bei Kindern, die älter waren als 3 Monate und bei Erwachsenen lagen sie stets vor. — In den muskulären Arterien traten die verkalkten Anteile der inneren elastischen Membran makroskopisch als schwarze paarweise angeordnete „Kalkbänder” entlang den Rändern vorgebildeter Membranspalten auf. Bei ausgeprägter Calcinose konnte mit der angewandten Methode das gesamte Spaltensystem der inneren elastischen Membran dargestellt werden. Kalkbänder wurden in den muskulären Arterien der unteren Extremität bei allen 10–20 Jahre alten Verstorbenen vorgefunden und waren auch in den nachfolgenden Altersstufen stets vorhanden. Beim Zusammenfluß von Kalkbändern und polygonalen Kalkinkrustationen entstehen in der inneren elastischen Membran folienartige Kalkablagerungen („Kalkfolien”). Die Kalkbänder und Kalkfolien stellen Kristallisationspunkte für körnige Kalkablagerungen dar, die zumeist an ihrer äußeren, der Media zugekehrten Seite entstehen.
    Notes: Abstract Calcification patterns of the internal elastic membrane of the main pelvic arteries, lower limb arteries, brachial, splenic and renal arteries were demonstrated grossly by a modified von Kossa technique. In the elastic segment in the common and internal iliac arteries, the membrane calcification appeared as groups of roundish or polygonal incrustations. They were found frequently in newborns, and were always present in infants of more than three months, as well as in adults. In the muscular arteries, the calcified parts of the internal elastic membrane appeared grossly as pairs of bands (“calcific bands”) along the edges of the pre-existing gaps in this membrane. When calcification was pronounced, the whole pattern of the membrane gaps could be demonstrated by the method used in this study. Calcific bands were found in the muscular arteries of the lower limbs in all 10–20 year-old subjects, and were always present in the older age groups. The confluence of calcific bands or polygonal membrane incrustations found in the iliac arteries lead to sheet-like membrane calcification. The calcific bands and sheets represent crystallizing points for grain-like calcific deposits, which appear later on the medial surface.
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    Calcified tissue international 3 (1969), S. 363-365 
    ISSN: 1432-0827
    Keywords: Calcification ; Mitochondria ; Protozoa
    Source: Springer Online Journal Archives 1860-2000
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    Calcified tissue international 3 (1969), S. 38-54 
    ISSN: 1432-0827
    Keywords: Epiphyseal cartilage ; Calcification ; Apatite crystals ; Organic/Inorganic relationships
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les rapports entre composants organiques et inorganiques du cartilage ont été étudiés au niveau des épiphyses tibiales chez des rats de 5 jours et au niveau des jonctions costo-chondrales chez des cobayes d'un mois. Voici les principaux résultats auxquels on est parvenu: (a) Dans les zones où la calcification s'amorce les cristallites d'apatite sont susceptibles d'être facilement éliminés des coupes en araldite et en glycol methacrylate (GMA) après traitement pendant une heure par une solution d'acide formique au 2%. (b) Les zones décalcifiées sont douées d'une faible densité aux électrons et vont exemptes d'une structure quelconque. Seulement la fixation à l'osmium peut révéler du matériel amorphe. (c) Indépendemment du fixateur et du moyen d'inclusion, les coupes décalcifiées, traitées par l'acétate d'uranyl et/ou par le citrate de plomb, montrent de fines structures allongées dont l'aspect est semblable à celui des cristallites. (d) Ces structures ainsi qu'un fond dense interposé prennent l'acide phosphotungstique (PTA) dans les coupes en GMA, une méthode celle-ci qui met en évidence les polysaccharides. (e) Le fond dense ne prend plus le PTA après traitement par la hyaluronidase et en même temps les structures semblables au cristallites apparaissent plus faibles. Après traitement par l'acétate d'uranyl et le citrate de plomb, l'attaque par la hyaluronidase ne produit aucune modification appréciable. Au contraire l'aspect des structures semblables aux cristallites apparait modifié par la digestion à la papaïne. (f) Quand la décalcification à l'EDTA précéde l'enrobage, des amas de matériel fibrillaire se mettent en évidence dans les zones où la calcification s'était amorcée. L'ensemble de ces résultats permet d'établir la nature organique des structures semblables au cristallites et porte aussi à admettre que celles-ci soient formées par de fins filaments cylindriques (digérés par la papaïne) entourés, et probablement engainés par del polysaccharides (digérés par la hyaluronidase et prennant le PTA dans les coupes en GMA).
    Abstract: Zusammenfassung Die Wechselbeziehungen zwischen den organischen und inorganischen Substanzen des verkalkenden Knorpels wurden in der tibialen und femuralen Epiphyse 5 Tage alter Ratten und in den Rippen 1 Monat alter Meerschweinchen untersucht. Die Hauptergebnisse waren: a) Die Hydroxyapatitkristalle der Frühverkalkungszonen können in Araldit- wie auch in Glycolmethylacrilatschnitten (GMA) mit einer lstündigen Behandlung durch 1% ige Ameisensäure leicht entfernt werden. b) Die entkalkten Zonen haben eine sehr niedrige Elektronendichte und enthalten keine sichtbaren Festkörper. Nach Fixierung mit Osmiumsäure kann man darin ein formloses Material erkennen. c) Eine Färbung der Schnitte mit Uranacetat und Bleicitrat, welche von den Fixier- und Einbettungsmitteln nicht beeinträchtigt wird, zeigt feine, längliche Körper (“Kristallgeister”) an, welche morphologisch den Kristallen von Hydroxyapatit gleichen. d) Die Färbung der GMA-Schnitte mit Phosphowolframsäure (PWS), eine Methode welche Polysaccharide anzeigt, hob diese “Kristallgeister” besonders hervor und demonstrierte auch eine dichte dazwischenliegende Grundsubstanz. e) Diese Substanz ist nach Behandlung mit Hyaluronidase nicht mehr färbbar mit PWS; die Färbbarkeit der “Kristallgeister” wird dabei jedoch nur verringert. Die Hyaluronidasebehandlung verändert die Uranacetat- und Bleicitratfärbung nicht. Die Feinstruktur der “Kristallgeister” wird durch Papainbehandlung sehr angegriffen. f) Falls die Schnitte vor dem Einbetten mit EDTA entkalkt werden, können Faserbündel in den Frühzonen der Verkalkung nachgewiesen werden. Diese Ergebnisse bestätigen die organische Natur der “Kristallgeister”. Sie beweisen auch, daß dieselben aus feinen Proteinstäbchen bestehen (verdaulich in Papain) und von Polysacchariden umgeben und vielleicht auch überzogen sind (verdaulich in Hyaluronidase und anfärbbar mit PWS).
    Notes: Abstract The organic/inorganic relationships in calcifying cartilage have been studied in tibialfemoral epiphyses of 5-day-old rats and in costochondral junctions of 1-month-old guinea pigs. The main results are: (a) Apatite crystallites in areas of early cartilage calcification are easily removed from araldite and glycol methacrylate (GMA) sections by 1-hour treatment with 2% formic acid. (b) The decalcified areas have a very low electron density and do not contain recognizable structures. Only after osmium fixation can an amorphous material be found in them. (c) Independently from the fixative and the embedding medium, staining decalcified areas with uranyl acetate and/or lead citrate reveals thin, elongated structures (crystallite ghosts) similar in shape to apatite crystallites. (d) These crystallite ghosts and a dense background are stained by phosphotungstic acid (PTA) in GMA sections, a method which reveals polysaccharides. (e) The dense background is no longer stained by PTA after hyaluronidase digestion and the stainability of the crystallite ghosts is reduced. After uranyl acetate/lead citrate staining there are no recognizable changes due to hyaluronidase. But, on the other hand, the fine structure of the crystallite ghosts is modified by papain digestion. (f) Clusters of fibrillar material are present in the areas of early calcification when EDTA decalcification is carried out before embedding the specimens. These results confirm the organic nature of the crystallite ghosts and suggest that they are formed by thin, rod-like protein/like structures (digested by papain) surrounded, and probably sheathed, by acid polysaccharides (digested by hyaluronidase and stained by PTA in GMA sections).
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    Calcified tissue international 4 (1969), S. 136-146 
    ISSN: 1432-0827
    Keywords: Bone ; Calcification ; Osteoblasts ; Osteoclasts ; Poultry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'activité des cellules osseuses et la composition du fémur de pondeuses furent examinées pendant sept jours de déficience calcique (diète contenant 0,13% de calcium) et sept jours de réplétion (diète contenant 3,2% de calcium). Du point de vue histologique, seul l'os cortical donnait des signes nets de résorption et d'activité ostéoclastique. Le nombre d'ostéoclastes dans l'os médullaire différait peu des valuers témoin pendant les périodes de déficience et de réplétion subséquente, sauf pour une augmentation significative au premier jour de déplétion. L'effect histologique le plus important dans l'os médullaire était une augmentation marquée en nombre d'ostéoblastes aux troisième, cinquième, et un peu moins au septième jours de déplétion. Le nombre d'ostéoblastes était en corrélation positive avec la teneur de l'os médullaire en ostéoide et négative avec son degré de calcification. L'activité de l'os médullaire en phosphatase alcaline augmentait avec la longueur de la déficience calcique. Un jour après le retour des pondeuses à une diète contenant 3,2% de calcium, la calcification de l'os médullaire avait augmenté de façon significative, le nombre d'ostéoblastes avait diminué au niveau ou au-dessous du niveau de contrôle et l'activité de la phosphatase alcaline avait baissé considérablement. L'importance de ces résultats est discutée par rapport au controle des populations des cellules dan l'os et au rôle de l'os médullaire.
    Abstract: Zusammenfassung Die Zahl der Knochenzellen und die Zusammensetzung des Femurs von Legehennen wurden während einer siebentägigen Calciumentzugsperiode (Calciumgehalt des Futters 0,13%) und einer siebentägigen Ersatzperiode (Calciumgehalt des Futters 3,2%) untersucht. Histologisch zeigte nur die Cortex eindeutige Knochenresorption und osteoklastische Aktivität. Abgesehen von einer signifikanten Zunahme am 1. Tag des Calciumentzuges, variierte die Zahl der Osteoklasten im Markknochen sowohl während der Entzugs- als auch während der nachfolgenden Ersatzperiode wenig. Die wichtigste histologische Änderung im Markknochen bestand in einer starken Zunahme in der Zahl der Osteoblasten am 3., 5. und etwas weniger am 7. Tag der Entzugsperiode. Die Zahl der Osteoblasten zeigte eine positive Korrelation mit dem Osteoidgehalt des Markknochens und eine negative mit dem Grade seiner Verkalkung. Die Aktivität der alkalischen Phosphatase im Markknochen war desto größer je länger den Hennen die calciumarme Ration verfüttert worden war. Die Wiederverabreichung der Ration, welche 3,2% Calcium enthielt, verursachte innerhalb eines Tages eine signifikante Zunahme in der Verkalkung des Markknochens, ein Absinken der Osteoblastzahl auf die Kontrollwerte oder unter sie und eine drastische Verringerung der alkalischen Phosphataseaktivität. Die Bedeutung dieser Ergebnisse in bezug auf die Kontrolle des Knochenzellenbestandes und auf die Funktion des Markknochens wird diskutiert.
    Notes: Abstract Bone cell activity and the composition of the femur of laying hens were studied during 7 days of calcium depletion on a 0.13% calcium diet and 7 days of calcium repletion on a 3.2% calcium diet. Histologically, only cortical bone showed clear signs of bone resorption and osteoclastic activity during the depletion period. The number of osteoclasts in medullary bone varied little from control values throughout both calcium depletion and repletion, except for a significant increase on the first day of depletion. The major histologicalchange in medullary bone was a marked increase in the number of osteoblasts on the third, fifth and, to a lesser extent, seventh, day of depletion. The number of osteoblasts in medullary bone was positively correlated with its osteoid content and negatively correlated with its degree of calcification. Alkaline phosphatase activity of medullary bone increased with the time the hens had been on the calcium-deficient diet. Returning the hens to the 3.2% calcium ration caused, within one day, a significant increase in medullary bone calcification, a decrease of osteoblast numbers to, or below, control levels, and a drastic reduction in alkaline phosphatase activity of medullary bone. The significance of these findings in relation to the control of bone cell populations and the functions of medullary bone is discussed.
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    Calcified tissue international 4 (1969), S. 101-112 
    ISSN: 1432-0827
    Keywords: Calcification ; Ligament ; Mollusc ; Aragonite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Une étude de microscopie électronique est réalisée sur la formation et la structure du ligament interne deMytilus edulis etPinctada radiata. Cette partie du ligament est dérivée des cellules isthmiques qui sont de forme cylindrique irrégulière. Elles présentent un ergastoplasme bien développé et un appareil de Golgi, engagé dans l'élaboration de vésicules et granules qui s'incorporent au niveau de la conchioline. Les cristaux se forment au niveau du front de calcification, à la surface interne du ligament. Ils sont entourés par une enveloppe. Ils se présentent comme des monocristaux d'aragonite, allongés et en forme d'aiguilles, dispersés dans le ligament. Bien que les constituants de la carapace et du ligament soient identiques, il existe des différences concernant l'augmentation quantitative de conchioline et une diminution en nombre, forme diverse, groupement et croissance des cristaux. Ces différences sont probablement liées à la fonction spécialisée du ligament.
    Abstract: Zusammenfassung Die Bildung und Struktur des inneren Ligamentes vonMytilus edulis undPinctada radiata wurden am Elektronenmikroskop untersucht. Dieser Teil des Ligamentes stammt von den Isthmuszellen ab, deren Form unregelmäßig säulenartig ist. Sie zeigen ein vorspringendes, rauhes endoplasmatisches Reticulum und einen Golgiapparat, welche sich mit der Bildung von Bläschen und Granula befassen, die schließlich in einem integralen Teil des Conchiolins eingebaut werden. Die Kristalle entstehen an der Calcifikationsgrenze an der inneren Oberfläche des Ligamentes und sind in Hüllen eingeschlossen. Sie bestehen aus langen, nadelförmigen, einzelnen Aragonit-Kristallen, die über das ganze Ligament verteilt sind. Obschon die Bestandteile der Muschel und des Ligamentes gleichartig sind, unterscheiden sich die beiden durch eine erhöhte conchiolinmenge, wie auch durch eine Abnahme der Anzahl der Kristalle, welche verschieden in der Form, in der Anordnung und im Wachstum sind. Dies alles ist vermutlich auf die spezielle Funktion des Ligamentes zurückzuführen.
    Notes: Abstract An electron microscope study was made of the formation and structure of the inner ligament ofMytilus edulis andPinctada radiata. This part of the ligament is derived from the isthmus cells which are irregular columnar in shape. They exhibit a prominent rough endoplasmic reticulum and a Golgi apparatus, which are concerned with the elaboration of vesicles and granules eventually incorporated into an integral part of the conchiolin. The crystals arise at the calcification front at the inner surface of the ligament and are enclosed in envelopes. They consist of long, needle-shaped, single aragonite crystals widely dispersed in the ligament. Although the components of the shell and ligament are similar, differences between them consist of an increased amount of conchiolin, as well as a decrease in the amount, diversity of form, arrangement and growth of the crystals; all probably related to the specialized function of the ligament.
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    Calcified tissue international 4 (1969), S. 359-365 
    ISSN: 1432-0827
    Keywords: Metals ; Ions ; Buffers ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'action de divers ions métalliques polyvalents sur les propriétés métastables d'un tampon calcifiable a été étudiée. A des concentrations de 1 μM ou moins, les ions Pb2+, F2+, Fe3+, Cr3+, Bi3+ et Al3+ déstabilisent le tampon. Il semble que les phosphates insolubles formés par ces ions déclanchent la précipitation de l'apatite. L'importance de ces résultats dans l'étiologie des calcifications pathologiques et la calciphylaxie est décrite.
    Abstract: Zusammenfassung Die Wirkung von verschiedenen polyvalenten Metallionen auf die Metastabilität eines Calcifikationspuffers wurde untersucht. Bei Konzentrationen von 1 μM oder weniger wird der Puffer durch Pb2+, Fe2+, Fe2+, Cr3+, Bi3+ und Al3+ entstabilisiert. Die Resultate lassen vermuten, daß unlösliche Phosphate, die mit diesen Ionen entstehen, eine Fällung von Apatit veranlassen. Die Bedeutung dieser Befunde bei pathologischen Verkalkungen und Calciphylaxis werden besprochen.
    Notes: Abstract The effect was studied of various polyvalent metal ions on the metastability of a calcification buffer. At concentration of 1 μM or less, Pb2+, Fe2+, Fe3+, Cr3+, Bi3+ and Al3+ ions were found to destabilize the buffer. The results suggest that insoluble phosphates formed by these ions initiate the precipitation of calcium apatite. The implications of these findings to pathological calcification and calciphylaxis are discussed.
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    Calcified tissue international 4 (1969), S. 39-47 
    ISSN: 1432-0827
    Keywords: Dehydroepiandrosterone ; Calcification ; Embryo ; Tissue Culture ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les modes d'utilisation de glucose, le contenu de calcium et d'hydroxyproline et la densité cellulaire du perioste de les os frontaux d'embryons de poulet de 12 et 13 jours de developpement, cultivés sur coagulum de plasma, se presentant différenment à chaque âge. Cultivés avec sulfate de déhydroèpiandrostérone en concentration 1 mM, les frontaux de 12 jours montrent un synthese augmentée du matrice osseuse, celle de 13 jours se calcifient à une vélocité significativement plus grande que celle des os contôles. Le degré de calcification au quatrième jour de culture measuré par la relation calcium/hydroxyproline, suit un fonction lineáire avec le logarithme des doses de sulfate de dehydroepiandrostérone employées (0.5, 1,0 et 2,0 mM). Les renseignements obtenus indiquent que les frontaux de 13 jours, cultivés “in vitro” constituent modeles experimentaux appropriés pour étudier l'effet des androgénes sur le tissue osseux.
    Abstract: Zusammenfassung Stirnbeine von Hühnerembryonen an ihrem 12. und 13. Entwicklungstag entnommen und in vitro kultiviert zeigen verschiedene Arten der Glucoseverwertung der Periostzellendichte, des Calcium- und Hydroxyprolingehaltes. Wird Dehydroepiandrosteronsulfat dem Medium in einer 1 mM-Konzentration zugegeben, so beteiligen sich die 12tägigen Stirnbeine vorwiegend an der Knochengewebesynthese, während die 13tägigen signifikant stärker verkalken als die Kontrollen. Gemessen an der Calcium/hydroxyprolin Ratio bildet die Verkalkung der 13tägigen Stirnbeine eine lineare Funktion mit den Logarithmen der verwendeten Dosen von Dehydroepiandrosteronsulfat (0,5, 1,0 und 2,0 mM). Das in vitro kultivierte 13tägige Stirnbein schein ein geeignetes Experimentiermodell zur Studie der Dehydroepiandrosteronsulfatwirkung auf das Knochengewebe zu sein, weil es das grundlegende Phänomen (erhöhte Verkalkung) wiedergibt, welches man auch bei mit Androgenen behandelten Menschen und Tieren beobachtet.
    Notes: Abstract Chick embryo frontal bones at 12 and 13 days of development cultivatedin vitro exhibit different patterns of glucose utilization, periosteal cellular density and calcium and hydroxyproline content. When dehydroepiandrosterone sulfate is added to the medium at a concentration 1 mM, 12-day frontals engage primarily in osteoid tissue synthesis while 13-day frontals calcify at a significantly greater rate than controls. Measured with the ratio calcium/hydroxyproline, the calcification of 13-day frontals follows a linear function with the logarithm of the doses of dehydroepiandrosterone sulfate employed (0.5, 1.0 and 2.0 mM). The 13-day frontal bone cultivatedin vitro seems to be an adequate experimental model for the study of the effects of dehydroepiandrosterone sulfate on bone tissue because it reproduces the basic phenomenon (increased calcification) observed in man and animals treated with androgens.
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  • 19
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    Calcified tissue international 56 (1995), S. 140-144 
    ISSN: 1432-0827
    Keywords: Bone sialoprotein ; Collagen ; Hole zone ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Bone sialoprotein (BSP) has an affinity to collagen fibrils [25]. A role of carbohydrate chains in the affinity was examined by removing sialic acids of BSP. Neuraminidase treatment of the BSP increased the binding to collagen. Binding sites of BSP on collagen were examined by biochemical and electron-microscopic methods. Purified bovine BSP was labeled with biotin. Collagen α chains or CNBr peptides were separated by electrophoresis and transfered to nitrocellulose membranes. The membranes were incubated with the biotin-labeled BSP, and the bound BSP was visualized with avidin conjugated with alkaline phosphatase. The labeled BSP was preferentially bound to the α 2 chain, and peptides derived from α 2 chain. In another experiment, the labeled BSP was incubated with reconstituted native collagen fibrils. The mixture was put on a copper grid, reacted with avidin conjugated with gold particles, and observed with an electron microscope. The gold particles were seen mainly within hole zones of the fibrils. BSP bound to the α 2 chain within the hole zones may regulate the onset of calcification at hole zones and the cell binding to collagen fibrils.
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  • 20
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    Calcified tissue international 2 (1968), S. 1-19 
    ISSN: 1432-0827
    Keywords: Collagen ; Hydroxyapatite ; Keratin ; Enamel ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
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  • 21
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    Calcified tissue international 2 (1968), S. 60-66 
    ISSN: 1432-0827
    Keywords: Fluorides ; Topical/Pharmacodynamics ; Penicillin/Pharmacodynamics ; Calcification ; Physiologic/Drug Effects ; Rats
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'objet de ce travail était de comparer le phénomène de la maturation postéruptive dans les molaires des rats ordinaires et sans germes à qui on a donné à manger un régime «non-cariogène». On a étudié les effets de l'application topique du fluorure et de la nourriture de la penicilline sur la maturation. Les molaires des rats ordinaires (nourris d'un régime «chow» — aliment mixte pour les animaux préparé d'habitude sous la forme d'une farine ou en boulettes) traités topiquement avec une solution à 1% de NaF et les molaires des rats ordinaires nourris d'un régime «chow» complété avec la penicilline à 1% ont montré un degré de maturation significativement plus grand que les molaires des rats de la même portée nourris de «chow» et traités avec de l'eau. De l'autre côté, les molaires des rats sans germes traités topiquement avec une solution à 1% de NaF n'ont montré aucune différence significative dans le degré de maturation des molaires de rats de contrôle semblables traités topiqument avec l'eau. Ces résultats sont en accord avec l'hypothèse que dans un milieu normal le procédé de minéralisation (maturation) est opposé par un procédé de déminéralisation. Le procédé de déminéralisation est un résultat de la production d'acide par des bactéries qui métabolisent les aliments encastrés dans les «sulci» des molaires. On propose que dans les animaux ordinaires le fluorure et la penicilline peuvent influencer la maturation en empêchant le procédé de déminéralisation. Dans les animaux sans germes le procédé de déminéralisation est absent parce que la microflore orale est absente.
    Abstract: Zusammenfassung Der Zweck dieser Arbeit war ein Vergleich des nach dem Durchbruch auftretenden Reifungsphänomens der Backenzähne von gewöhnlich und von keimfrei gehaltenen Ratten, die mit einer nicht-cariogenen Diät ernährt wurden. Es wurden die Auswirkungen von topisch angewendetem Fluorid, verbunden mit Penicillin-Fütterung auf die Reifung untersucht. Die Backenzähne der gewöhnlich gehaltenen Ratten (mit “Chow”-Diät ernährt), die mit einer 1%igen Na-Fluoridlösung topisch behandelt wurden, und solchen deren “Chow”-Diät zu 1% mit Penicillin versetzt wurde, zeigten einen erheblich größeren Reifungsgrad als die Backenzähne von Tieren des gleichen Wurfes, die nur mit der “Chow”-Diät und Wasser ernährt wurden. Diese Resultate stimmen mit der Hypothese überein, daß in einer normalen Umgebung dem Mineralisationsprozeß (Reifung) ein Demineralisationsprozeß entgegenwirkt. Der Demineralisationsprozeß ist durch eine Säureproduktion von Bakterien bedingt, welche die in die Zahnfurchen eingepreßten Nahrungsbestandteile metabolisieren. Es wird die Annahme vorgeschlagen, daß Fluoride und Penicillin bei den gewöhnlich gehaltenen Tieren die Reifung durch eine Inhibition des Demineralisationsprozesses beeinflussen. Bei keimfrei gehaltenen Tieren findet dagegen kein Demineralisationsprozeß statt, da die orale Mikroflora fehlt.
    Notes: Abstract The purpose of this work was to compare the phenomenon of post-eruptive maturation in molars of conventional and germfree rats fed non-cariogenic diets. The effects of topical application of fluoride and feeding penicillin on maturation were studied. The molars of conventional rats (fed a chow diet) treated topically with a 1% NaF solution and the molars of conventional rats fed a chow diet supplemented with 1% penicillin showed a significantly greater degree of maturation than did the molars of littermate rats fed chow and treated with water. On the other hand, the molars of germfree rats treated topically with a 1% NaF solution showed no significant difference in degree of maturation from the molars of similar control rats topically treated with water. These data are consistent with an hypothesis that in a normal environment the mineralization (maturation) process is opposed by a demineralization process. The demineralization process is a result of production of acid by bacteria metabolizing the diet impacted in the sulci of molars. It is proposed that in conventional animals fluoride and penicillin may influence maturation by inhibiting the demineralization process. In the germfree animals the demineralization process is absent because the oral microflora is absent.
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  • 22
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    Calcified tissue international 2 (1968), S. 361-369 
    ISSN: 1432-0827
    Keywords: Calcification ; Hydroxyapatite ; Keratin ; Hair ; Electron diffraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Certaines régions du cortex des fibres de kératine contiennent une proportion relativement élevée de matériel non kératinisé. L'orientation microfibrillaire dans ces régions est biaxiale et on y trouve, en outre, des zones de matériel amorphe. Les sels calciques, et en particulier l'hydroxyleapatite, sont étudiés par diffraction électronique qui permet de les localiser et de les identifier. En outre, cette technique permet de déterminer la fréquence de cellules nonkératinisées dans la partie interne du cortex et de les comparer avec les propriétés histochimiques de cette région. Une comparison est effectuée entre la calcification de la kératine et celle intéressant le collagène et l'élastine.
    Abstract: Zusammenfassung Es wird gezeigt, daß Bereiche im Cortex von gewissen Keratinfasern einen relativ hohen Anteil an nichtkeratinösem Material enthalten. Ferner konnte nachgewiesen werden, daß die Orientierung der Mikrofibrillen in diesen Bereichen biaxialer Art ist, und daß Bezirke von amorphem Material vorhanden sind. Die Calciumsalze und insbesondere das Hydroxyapatit konnten mit Hilfe der Elektronendiffraktion lokalisiert und identifiziert werden. Diese Methode hat sich auch als wertvoll erwiesen, um die Wechselbeziehung zwischen dem Vorkommen von einigen nichtkeratinisierten Zellen im Cortexinnern und den histochemischen Eigenschaften dieser Regionen zu zeigen. Ein Vergleich wird gemacht zwischen der Umgebung der Verkalkungsstellen im Keratin mit derjenigen im Kollagen und im Elastin.
    Notes: Abstract Regions in the cortex of some keratin fibres are shown to contain a relatively high proportion of non-keratinous material. The microfibrillar orientation in these regions is shown to be biaxial in nature and, in addition, areas of amorphous material are reported. Calcium salts, in particular hydroxyapatite, have been located and identified using electron diffraction, which has also proved valuable in correlating the incidence of some non-keratinised cells in the inner part of the cortex with the histochemical properties of this region. A comparison is made between the environment of the sites of calcification in keratin with those in collagen and elastin.
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  • 23
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    Calcified tissue international 2 (1968), S. 214-228 
    ISSN: 1432-0827
    Keywords: Calcinosis ; Calcification ; Cartilage ; Collagen ; Mineral metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les facteurs, influençant la vitesse et l'intensité du phénomène d'association des ions calcium et phosphates avec des fibres contenant du collagène, et préparés à partir du tendon de boeuf par deux méthodes d'extraction différentes, ont été étudiés. Les fibres, obtenues par ces deux méthodes, nécessitent spécifiquement du phosphate pour absorber du calcium et vice versa. L'absorption ionique des deux préparations est inhibée par du Mg++, du pyrophosphate et un peptide acidique, isolé du sérum humain. Alors que les fibres contenant du collagène, préparées selon les deux méthodes, présentent une absorption ionique à des vitesses sensiblement identiques, seule une des méthodes donne une matrice réagissant positivement à la technique de coloration au nitrate d'argent de von Kossa. Etant donné que les deux critères de calcification sont intéressés de façon identique par des conditions de réaction et par des inhibiteurs, il apparait que les deux facteurs sont des manifestations de différents stades de calcification et que des études d'absorption ionique fournissent une base quantitative d'appréciation de la calcification, pouvant être d'importance pour l'étude du mécanisme et de contrôle de la minéralisation tissulaire.
    Abstract: Zusammenfassung Überprüft wurden die Faktoren, welche Geschwindigkeit und Ausmaß der Erscheinung beeinflussen, wobei Calcium- und Phosphationen sich mit den kollagenhaltigen, durch zwei verschiedene Extraktionsmethoden aus Rindersehnen gewonnenen Fasern eng zusammenbinden. Die mit beiden Methoden zubereiteten Fasern benötigen spezifisch Phosphat für die Calciumaufnahme und Calcium für die Phosphataufnahme. Die Ionenaufnahme beider Arten wird durch Mg++, Pyrophosphat und saure, aus dem menschlichen Serum isolierte Peptide gehemmt. Während die nach beiden Methoden präparierten kollagenhaltigen Fasern eine Ionenaufnahme von beinahe gleicher Geschwindigkeit verursachen, ergibt nur eine dieser Methoden eine Matrix, die mit der Silbernitratfärbung nach vonKossa positiv reagiert. Da beide Calcifikationskriterien gleicherweise durch Reaktionsbedingungen und Inhibitoren beeinflußt werden, wird daraus geschlossen, daß beide Erscheinungen verschiedener Stadien des Gesamtcalcifikationsprozesses sind. Untersuchungen über die Ionenaufnahme ergeben eine quantitative Angabe der Verkalkung, welche für die Erforschung des Mechanismus und der Kontrolle der Mineralisation der Gewebe wichtig sein könnte.
    Notes: Abstract Factors that influence the rate and extent of the phenomenon in which calcium and phosphate ions become firmly associated with collagen-containing fibers prepared from beef tendon by two different extraction methods have been investigated. The fibers produced by both methods specifically require phosphate for calcium uptake and calcium is required for phosphate uptake. Ion uptake by both types is inhibited by Mg++, pyrophosphate, and an acidic peptide isolated from human serum. Whereas the collagen-containing fibers prepared by both methods induce ion uptake at nearly identical rates, only one of the methods produced a matrix that gives a positive response to the silver nitrate staining technique of von Kossa. Since both criteria of calcification are similarly influenced by reaction conditions and inhibitors, it is concluded that both are manifestations of different stages of the overall calcification process and that studies of ion uptake provide a quantitative assessment of calcification which could be of importance for investigating the mechanism and control of tissue mineralization.
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  • 24
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    Calcified tissue international 56 (1995), S. 403-407 
    ISSN: 1432-0827
    Keywords: Bone sialoprotein ; Fibronectin ; Type I colfagen ; MC3T3-E1 cells ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Bone sialoprotein (BSP) containing an Arg-Gly-Asp cell-binding sequence was purified from bovine bone 4 M guanidine-HCl extract after HCl demineralization by a series of chromatographic procedures. When this protein was coated on culture dishes in the presence of type I collagen, it increased both DNA content and alkaline phosphatase (ALP) activity in osteoblast-like MC3T3-E1 cells, and stimulated calcification in the cells, whereas fibronectin, another cell-binding protein, showed a marked increase in the DNA content but had little effect on the ALP activity. These findings suggest that BSP is mitogenic for preosteoblasts and differentiating the cells into osteoblasts, thereby stimulating bone calcification
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  • 25
    ISSN: 1432-1939
    Keywords: Key words Agrostis canina ; CO2 vents ; photosynthesis ; lignification ; growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The aim of this study was to characterise growth and photosynthetic capacity in plants adapted to long-term contrasting atmospheric CO2 concentrations (C a). Seeds of Agrostis canina L. ssp. monteluccii were collected from a natural CO2 transect in central-western Italy and plants grown in controlled environment chambers at both ambient and elevated CO2 (350 and 700 μmol mol−1) in nutrient-rich soil. Seasonal mean C a at the source of the plant material ranged from 610 to 451 μmol CO2 mol−1, derived from C4 leaf stable carbon isotope discrimination (δ13C). Under chamber conditions, CO2 enrichment stimulated the growth of all populations. However, plants originating from elevated C a exhibited higher initial relative growth rates (RGRs) irrespective of chamber CO2 concentrations and a positive relationship was found between RGR and C a at the seed source. Seed weight was positively correlated with C a, but differences in seed weight were found to explain no more than 34% of the variation in RGRs at elevated CO2. Longer-term experiments (over 98 days) on two populations originating from the extremes of the transect (451 and 610 μmol CO2 mol−1) indicated that differences in growth between populations were maintained when plants were grown at both 350 and 700 μmol CO2 mol−1. Analysis of leaf material revealed an increase in the cell wall fraction (CWF) in plants grown at elevated CO2, with plants originating from high C a exhibiting constitutively lower levels but a variable response in terms of the degree of lignification. In vivo gas exchange measurements revealed no significant differences in light and CO2 saturated rates of photosynthesis and carboxylation efficiency between populations or with CO2 treatment. Moreover, SDS-PAGE/ LISA quantification of leaf ribulose bisphosphate carboxylase/oxygenase (Rubisco) showed no difference in Rubisco content between populations or CO2 treatments. These findings suggest that long-term adaptation to growth at elevated CO2 may be associated with a potential for increased growth, but this does not appear to be linked with differences in the intrinsic capacity for photosynthesis.
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  • 26
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    Water, air & soil pollution 85 (1995), S. 1461-1466 
    ISSN: 1573-2932
    Keywords: ozone ; wheat ; Triticum aestivum ; growth ; senescence ; biomass partitioning ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract In closed-chamber fumigation experiments dry matter partitioning and chlorophyll fluorescence of wheat were studied, analysing the effects of ozone during different stages of plant development. Ozone causes enhanced leaf senescence, leading to a loss of green leaf area and, consequently to a decreased supply of assimilates, affecting (in increasing order of severeness) stem, ear and grain productivity because of reduced storage pools for translocation. Leaves of plants before shooting stage were most sensitive but the lack of green leaf area after ear emergence had the most pronounced effects on grain yield. Measurements of photochemical capacity showed that evidence for negative ozone effects could be found in changes of chlorophyll fluorescence parameters in leaf sections not yet showing visible ozone injury. Negative effects on photosynthesis were more distinct with increasing accumulated ozone dose, with increasing age of leaf tissue and with increasing ozone sensitivity of the cultivar. The changes in chlorophyll fluorescence are most likely to be explained by a decreased pool size of plastoquinones caused by ozone.
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  • 27
    ISSN: 1573-5028
    Keywords: Cell cycle ; circadian clock ; green alga ; GTP-binding proteins ; light regulation ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract ADP-ribosylation factor (ARF) is a highly conserved, low molecular mass (ca. 21 kDa) GTP-binding protein that has been implicated in vesicle trafficking and signal transduction in yeast and mammalian cells. However, little is known of ARF in plant systems. A putative ARF polypeptide was identifed in subcellular fractions of the green alga Chlamydomonas reinhardtii, based on [32P]GTP binding and immunoblot assays. A cDNA clone was isolated from Chlamydomonas (Arf1), which encodes a 20.7 kDa protein with 90% identity to human ARF1. Northern blot analyses showed that levels of Arf1 mRNA are highly regulated during 12 h/12 h light/dark (LD) cycles. A biphasic pattern of expression was observed: a transient peak of Arf1 mRNA occurred at the onset of the light period, which was followed ca. 12 h later by a more prominent peak in the early to mid-dark period. When LD-synchronized cells were shifted to continuous darkness, the dark-specific peak of Arf1 mRNA persisted, indicative of a circadian rhythm. The increase in Arf1 mRNA at the beginning of the light period, however, was shown to be light-dependent, and, moreover, dependent on photosynthesis, since it was prevented by DCMU. We conclude that the biphasic pattern of Arf1 mRNA accumulation during LD cycles is due to regulation by two different factors, light (which requires photosynthesis) and the circadian clock. Thus, these studies identify a novel pattern of expression for a GTP-binding protein gene.
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  • 28
    ISSN: 1573-5028
    Keywords: gene expression ; photosynthesis ; protein turnover ; psbA ; tac promoter ; D1 protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Over-expression of the psbAIII gene encoding for the D1 protein (form II; D1:2) of the photosystem II reaction centre in the Synechococcus sp. PCC 7942 was studied using a tac promoter and the lacI Q system. Over-expression was induced with 40 μg/ml IPTG in the growth medium for either 6 or 12 h at growth irradiance (50 μmol photons m-2 s-1). This treatment doubled the amount of psbAII/III mRNA and the D1:2 protein in membranes but decreased the amount of psbAI messages and the D1:1 protein. The total amount of both heterodimeric reaction centre proteins, D1 and D2, remained constant under growth light conditions, indicating that the number of PSII centres in the membranes was not affected, only the form of the D1 protein was changed from D1:1 to D1:2 in most centres. When the cells were photoinhibited either at 500 or 1000 μmol photons m-2 s-1, in the presence or absence of the protein synthesis inhibitor lincomycin, the D1:2 protein remained at a higher level in cells in which over-expression had been induced by IPTG. These cells were also less prone to photoinhibition of PSII. It is suggested that the tolerance of cells to photoinhibition increases when most PSII reaction centres contain the D1:2 protein at the beginning of high irradiance. This tolerance is further strengthened by maintaining psbAIII gene over-expression during the photoinhibitory treatment.
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  • 29
    ISSN: 1573-5028
    Keywords: antisense ; Corynebacterium glutamicum ; Escherichia coli ; Flaveria trinervia ; overexpression ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phosphoenolpyruvate carboxylase (PEPC) genes from Corynebacterium glutamicum (cppc), Escherichia coli (eppc) or Flaveria trinervia (fppc) were transferred to Solanum tuberosum. Plant regenerants producing foreign PEPC were identified by Western blot analysis. Maximum PEPC activities measured in eppc and fppc plants grown in the greenhouse were doubled compared to control plants. For cppc a transgenic plant line could be selected which exhibited a fourfold increase in PEPC activity. In the presence of acetyl-CoA, a known activator of the procaryotic PEPC, a sixfold higher activity level was observed. In cppc plants grown in axenic culture PEPC activities were even higher. There was a 6-fold or 12-fold increase in the PEPC activities compared to the controls measured in the absence or presence of acetyl-CoA, respectively. Comparable results were obtained by transient expression in Nicotiana tabacum protoplasts. PEPC of C. glutamicum (PEPC C.g.) in S. tuberosum leaf extracts displays its characteristic K m(PEP) value. Plant growth was examined with plants showing high expression of PEPC and, moreover, with a plant cell line expressing and antisense S. tuberosum (anti-sppc) gene. In axenic culture the growth rate of a cppc plant cell line was appreciably diminished, whereas growth rates of an anti-sppc line were similar or slightly higher than in controls. Malate levels were increased in cppc plants and decreased in antisense plants. There were no significant differences in photosynthetic electron transport or steady state CO2 assimilation between control plants and transformants overexpressing PEPC C.g. or anti-sppc plants. However, a prolonged dark treatment resulted in a delayed induction of photosynthetic electron transport in plants with less PEPC. Rates of CO2 release in the dark determined after a 45 min illumination period at a high proton flux density were considerably enhanced in cppc plants and slightly diminished in anti-sppc plants. When CO2 assimilation rates were corrected for estimated rates of mitochondrial respiration in the light, the electron requirement for CO2 assimilation determined in low CO2 was slightly lower in transformants with higher PEPC, whereas transformants with decreased PEPC exhibited an appreciably elevated electron requirement. The CO2 compensation point remained unchanged in plants (cppc) with high PEPC activity, but might be increased in an antisense plant cell line. Stomatal opening was delayed in antisense plants, but was accelerated in plants overexpressing PEPC C.g. compared to the controls.
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  • 30
    ISSN: 1573-5028
    Keywords: photosystem II ; photosynthesis ; chlorophyll-binding protein ; Synechocystis ; oxygen evolution ; oligonucleotide-mediated mutagenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract CP 47, a component of photosystem II (PSII) in higher plants, algae and cyanobacteria, is encoded by the psbB gene. Site-specific mutagenesis has been used to alter a portion of the psbB gene encoding the large extrinsic loop E of CP 47 in the cyanobacterium Synechocystis 6803. Alteration of a lysine residue occurring at position 321 to glycine produced a strain with altered PSII activity. This strain grew at wild-type rates in complete BG-11 media (480 µM chloride). However, oxygen evolution rates for this mutant in complete media were only 60% of the observed wild-type rates. Quantum yield measurements at low light intensities indicated that the mutant had 66% of the fully functional PSII centers contained in the control strain. The mutant proved to be extremely sensitive to photoinactivation at high light intensities, exhibiting a 3-fold increase in the rate of photoinactivation. When this mutant was grown in media depleted of chloride (30 µM chloride), it lost the ability to grow photoautotrophically while the control strain exhibited a normal rate of growth. The effect of chloride depletion on the growth rate of the mutant was reversed by the addition of 480 µM bromide to the chloride-depleted BG-11 media. In the presence of glucose, the mutant and control strains grew at comparable rates in either chloride-containing or chloride-depleted media. Oxygen evolution rates for the mutant were further depressed (28% of control rates) under chloride-limiting conditions. Addition of bromide restored these rates to those observed under chloride-sufficient conditions. Measurements of the variable fluorescence yield indicated that the mutant assembled fewer functional centers in the absence of chloride. These results indicate that the mutation K321G in CP 47 affects PSII stability and/or assembly under conditions where chloride is limiting.
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  • 31
    ISSN: 1573-5028
    Keywords: chloroplasts ; gene expression ; heat bleaching ; photosynthesis ; transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A shift in the ratio of chlorophyll (Chl) a and Chl b is an early indicator of heat bleaching in Euglena gracilis. This observation prompted us to consider whether or not changes in steady-state levels of chloroplast transcripts and in transcriptional activity could limit the synthesis of Chl a-binding proteins in bleaching plastids. We found that the mature transcripts for CP47 and CP43, the Chl a-binding apoproteins of the proximal antenna of photosystem II, decline sharply very early during bleaching. Our study also shows that transcription of psbB and psbC, the chloroplast genes encoding CP47 and CP43, remains essentially unchanged during the same interval. We conclude that posttranscriptional events, such as mRNA stability, could play a major role in initiating an irreversible loss of chloroplast function in Euglena at a moderately elevated temperature. Lack of these transcripts would eventually impair the assembly of photosystem II in thylakoids.
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  • 32
    ISSN: 1573-5028
    Keywords: Cyanobacteria ; gene copy number ; light regulation ; photosynthesis ; photosystem II reaction center ; polymerase chain reaction ; psbA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract DNA sequence, copy number, expression and phylogenetic relevance of the psbA gene from the abundant marine prokaryote P. marinus CCMP 1375 was analyzed. The 7 amino acids near the C-terminus missing in higher plant and in Prochlorothrix hollandica D1 proteins are present in the derived amino acid sequence. P. marinus contains only a single psbA gene. Thus, this organism lacks the ability to adapt its photosystem II by replacement of one type of D1 by another, as several cyanobacteria do. Phylogenetic trees suggested the D1-1 iso-form from Synechococcus PCC 7942 as the next related D1 protein and place P. Marinus separately from Prochlorothrix hollandica among the cyanobacteria.
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  • 33
    ISSN: 1573-5028
    Keywords: chlorophyll synthesis ; cyanobacteria ; chlorophyl-binding proteins ; photosynthesis ; thylakoid membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Part of the chlL gene encoding a component involved in light-independent protochlorophyllide reduction was deleted in wild type and in a photosystem I-less strain of Synechocystis sp. PCC 6803. In resulting mutants, chlorophyll biosynthesis was fully light-dependent. When these mutants were propagated under light-activated heterotrophic growth conditions (in darkness except for 15 min of weak light a day) for several weeks, essentially no chlorophyll was detectable but protochlorophyllide accumulated. Upon return of the chlL - mutant cultures to continuous light, within the first 6 h chlorophyll was synthesized at the expense of protochlorophyllide at a rate independent of the presence of photosystem I. Chlorophyll biosynthesized during this time gave rise to a 685 nm fluorescence emission peak at 77 K in intact cells. This peak most likely originates from a component different from those known to be directly associated with photosystems II and I. Development of 695 and 725 nm peaks (indicative of intact photosystem II and photosystem I, respectively) required longer exposures to light. After 6 h of greening, the rate of chlorophyll synthesis slowed as protochlorophyllide was depleted. In the chlL - strain, greening occurred at the same rate at two different light intensities (5 and 50 μE m-2s-1), indicating that also at low light intensity the amount of light is not rate-limiting for protochlorophyllide reduction. Thus, in this system the rate of chlorophyll biosynthesis is limited neither by biosynthesis of photosystems nor by the light-dependent protochlorophyllide reduction. We suggest the presence of a chlorophyll-binding ‘chelator’ protein (with 77 K fluorescence emission at 685 nm) that binds newly synthesized chlorophyll and that provides chlorophyll for newly synthesized photosynthetic reaction centers and antennae.
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  • 34
    ISSN: 1573-5028
    Keywords: Cyanobacteria ; photosynthesis ; random mutagenesis ; sodium bisulfite ; thylakoids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To identify amino acid residues of the D2 protein that are critical for functional photosystem II (PS II), sodium bisulfite was utilized for in vitro random mutagenesis of the psbDI gene from Synechocystis sp. PCC 6803. Sodium bisulfite reacts specifically with cytosine in single-stranded regions of DNA and does not attack double-stranded DNA. Using a hybrid plasmid that was single-stranded in the region to be mutagenized and that was double-stranded elsewhere, mutations were targeted to a specific psbDI region coding for the lumenal A-B loop of the D2 protein. Several mutants were isolated with a total of 15 different amino acid changes in the loop. The majority of these mutations did not result in a loss of photoautotrophic growth or in significantly altered PS II function. However, mutation of Glu-69 to Lys, Ser-79 to Phe, and Ser-88 to Phe were found to influence photosystem II activity; the importance of the latter two residues for proper PS II function was unexpected. Cells carrying the double mutation S79F/S88F in D2 did not grow photoautotrophically and had no functionally active PS II centers. The single mutant S79F was also incapable of photoautrophic growth, but displayed reasonably stable oxygen evolution, while PS II function in the single mutant S88F appeared to be close to normal. Because of the more pronounced phenotype of the S79F/S88F strain as compared to the single mutants, both Ser residues appear to affect stable assembly and function of the PS II complex. The mechanism by which the S79F mutant loses photoautotrophic growth remains to be established. However, these results show the potential of targeted random mutagenesis to identify functionally important residues in selected regions of proteins.
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  • 35
    ISSN: 1871-4528
    Keywords: benzylaminopurine ; gibberellic acid ; senescence ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The modifying effects of applying the plant growth regulators (PGRs) benzylaminopurine (BAP), gibberellic acid (GA3) and BAP+GA3 on physiological age were studied. Two experiments with two cultivars, differing in rate of physiological ageing (medium-early Pampeana, medium-late Huinkul) and two storage systems were performed during 1988/89 and 1989/90 in two different potato areas of Argentina. In both seasons seed tubers stored in heaps reached an advanced physiological age at planting, compared with tubers from the cold store. Seed tubers of cv. Pampeana were older than those of Huinkul. compared with control crops, those sprayed with BAP maintained ground cover and photosynthesis for longer, and those sprayed with GA3 for a shorter period. Consequently tuber yield was decreased by GA3 in 1988/89, but in 1989/90 all crops treated with PGRs outyielded the control. BAP could overcome effects of advanced physiological age on crop senescence and tuber yield.
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  • 36
    ISSN: 1871-4528
    Keywords: water stress ; photosynthesis ; stress index ; dry matter partitioning ; tuber quality ; Solanum tuberosum L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A lysimeter experiment was performed to study the optimal allocation of limited water supply in potatoes. Irrigation regimes equal to 40, 60 and 80% of maximum evapotranspiration (ET) were evenly applied over the crop cycle. Other treatments involved withholding 80 mm of irrigation, based on ET, beginning at each of three designated growth stages (tuber initiation, early and late tuber growth). An irrigated control treatment, restoring the entire ET, was included for comparison. Continuous drought stress reduced photosynthesis as irrigation volumes were reduced. Plant biomass and tuber yield decreased almost proportionally to water consumption, so that WUE was roughly constant. N uptake was highest in the control and in 80% ET treatment. Withholding water during tuberisation severely hindered plant physiological processes and penalized tuber yield. Reductions in photosynthesis, total biomass and yield were the greatest when drought was imposed during tuber initiation. The earliest stress resulted in the lowest WUE and N uptake. A new crop water stress index (SI) was proposed, which combines atmospheric demand for water and canopy temperature.
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  • 37
    ISSN: 1610-7403
    Keywords: Chamaecyparis obtusa Endl. ; photosynthesis ; Pinus densiflora Sieb. et Zucc. ; stomatal conductance ; water stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract This study was conducted to investigate the potential for modifying drought tolerance of Japanese cypress (Chamaecyparis obtusa Endl.) and Japanese red pine (Pinus densiflora Sieb. et Zucc.). Three-year-old seedlings were controlled for five-months at three different soil water potentials ({ie73-1}). Japanese cypress exposed to high {ie73-2} was able to maintain higher photosynthesis (Phn), transpiration (Tr) and stomatal conductance to H2O (gH2O) in comparison to low {ie73-3} pretreatments, however, there was no significant difference in Phn for Japanese red pine. Soil water potential at the threshold from the maximum to limited Phn was higher in high {ie73-4} pretreatments than in low {ie73-5} pretreatments. Net photosynthesis, Tr and gH2O decreased more rapidly in high {ie73-6} pretreatments than in low {ie73-7} pretreatments. Transpiration decreased more significantly than Phn, thus, resulted in increased water use efficiency. All these factors are likely to result in significant improvements in the drought tolerance. Japanese red pine seems more drought-tolerant than Japanese cypress. Japanese cypress is suitable to soil of −0.05 MPa water potential, and Japanese red pine is suitable to −0.16 MPa and even dryer soils.
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  • 38
    ISSN: 1610-7403
    Keywords: chlorophyll fluorescence ; electron transport ; photorespiration ; photosynthesis ; quantum yield ; tree
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Using attached and detached leaves ofAcer palmatum Thunb. andRhaphiolepsis umbellata Makino, pulse-modulated chlorophyll fluorescence and CO2 exchange were measured. Quantum yield of photosynthesis was determined from the fluorescence parameter(Fm′−Fs)/Fm′, where (Fm′−Fs) was defined as the difference between steady state chlorophyll fluorescence (Fs) and maximum fluorescence (Fm′) elicited by a saturating light pulse. The rate of electron transport through photosystem II (total electron flow) was calculated from the product of quantum yield andA (PFD), whereA is the rate of absorbed photons as given by leaf absorptance, and PFD is the photon flux density at the leaf surface. The rate of electron transport dependant on CO2 uptake (assimilative electron flow) was calculated from the gross photosynthetic rate in a leaf. The difference between the rates of total and assimilative electron transport was denoted as the rate of non-assimilative electron transport which depends on photorespiration and oxygen reduction. Available data provided quantitative information on the rate of non-assimilative electron flow in intact leaves. When leaf photosynthesis ofA. palmatum was measured under sunlight, the rates of total and assimilative electron transport were determined to be approximately 900 and 150 μmol equiv. e/mg Chl·h, respectively. The difference (750 μmol equiv. e/mg Chl·h) was attributed to the activity of non-assimilative electron flow. The ratio of total to assimilative electron flow was found to increase gradually with rising in irradiance. The results suggest that non-assimilative electron flow occurred at much higher rate than assimilative electron flow at high irradiance. Implications of the results are briefly discussed in relation to photosynthesis limitation in tree leaves.
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  • 39
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    Chromatographia 40 (1995), S. 99-101 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; High temperature open-tubular columns ; Solvent make-up ; Acetophenone oxidation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Acetophnone, a substance stable at room temperature, is readily oxidizes in a high temperature open tubular liquid chromatographic system by the residual oxygen in the mobile phase. The oxidation product is less UV absorptive and detection sensitivity decreases greatly. To prevent the oxidation, through degassing of the mobile phase is necessary.
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  • 40
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Endotoxins ; Lipopolysaccharide ; Aminated poly(γ-methyl L-glutamate) ; Bovine serum albumin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method is described for the selective removal of endotoxins from various protein solutions using columns packed with aminated poly (γ-methyl L-glutamate) (PMLG-NH2) spheres. The PMLG-NH2 adsorbents showed a high adsorbing activity for endotoxins which had an ionic strength of μ=0.05–1.0 and pH 5.0–9.0. The endotoxin-adsorbing capacity per millitre of the wet adsorbent increased from 0.40 to 1.35 mg (E. coli O111: B4 LPS) at μ=0.2 and pH 7.0 while the aminogroup content of the adsorbent increased from 0.8 to 3.5 meq g−1. The PMLG-NH2-3.5 has an amino-group content of 3.5 meq g−1. This column packing selectively adsorbed endotoxins, without loss of the protein, from a γ-globulin or cytochromec solution which contained endotoxins at μ=0.05 and pH 7.0. On the other hand, when bovine serum albumin (BSA) was present in solution with endotoxins, both the endotoxins and the BSA were adsorbed by the column. The BSA-adsorbing activity increased with increasing amino-group content of the adsorbent. However, this undesirable adsorption was suppressed with increasing ionic strength of the buffer. As a result, when the packing which had an amino-group content of 1.5 meg g−1 was used in conditions of μ=0.2 and pH 7.0, the endotoxins were removed from a BSA-containing solution without affecting the recovery of the BSA.
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  • 41
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    Chromatographia 40 (1995), S. 51-57 
    ISSN: 1612-1112
    Keywords: Column electrophoresis ; Column liquid chromatography ; Phenolic lignin degradation compounds ; Sugar degradation products ; Quantitative analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method for the quantitative analysis of phenolic lignin degradation products by capillary electrophoresis (CE) with on-column UV detection has been developed. The liquid biomass solutions contain low molecular hemicellulosic sugars and phenolic lignin degradation products with various degrees of polymerization. Special attention has been paid to the monomeric phenolic components of lignin degradation fragments, e.g. derivatives of phenolic acids, aldehydes, and alcohols. Uncoated fused silica capillaries and borate-phosphate buffer systems at moderate pH conditions were used in order to separate the compounds of interest. To provide validation of the method, the same samples were analyzed independently by HPLC using gradient elution on a RP-C18 column. As sugar degradation fragment, furan-2-carboxylic acid was detected.
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  • 42
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    Chromatographia 40 (1995), S. 85-90 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Phenolic compounds ; Ion-pair solid phase extraction ; On-line trace enrichment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Two packing materials, C18 and PLRP-S, are studied for on-line trace enrichment of phenolic compounds in water. Various precolumns of different internal diameter are also tested and the addition of an ion-pair reagent to increase retention and thus, breakthrough volumes of phenolic compounds, is studied. Best results are obtained when a PLRP-S precolumn is coupled on-line with a C18 analytical column and DAD detector. Addition of TBA considerably increases breakthrough volumes. In contrast, when a C18 precolumn is used, breakthrough volumes are lower and it is impossible to determine TCP and PCP, under the experimental conditions used, because of interference of other nonpolar compounds in the samples. The performance of the system is evaluated with river and tap water and the preconcentration of 10 ml of sample in a PLRP-S precolumn involves a linear range between 1 μg 1−1 and 20 μl−1 and limits of determination between 0.5 μg l−1 and 1 μg l−1 are obtained.
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  • 43
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    Chromatographia 40 (1995), S. 631-637 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Trace analysis ; Pesticides in tap water ; Aromatic sulphonates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Methodology and instrumentation are described which allow chromatographic screening of water samples under substantially simplified conditions and at reduced cost. A single column is used to accomplish sample extraction, trace enrichment, and separation. The performance of such a system is demonstrated, and the results compared with conventionally used concepts.
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  • 44
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Transchelation ; Zinc pyrithione and related compounds ; Antidandruff formulations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary An analytical procedure has been developed for the determination of zinc pyrithione in antidandruff formulations. Zinc pyrithione was converted into a stable copper complex and then analysed by reversed-phase HPLC. The proposed method allows the separation of the analyte from related pyrithiones and therefore is able to verify the compliance of cosmetic preparations with current legislation.
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  • 45
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Benzodiazepines ; Solvent effects ; ACN clusters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Two different methods were used to determine the separation factor α at different temperatures and the Gibbs-Helmholtz parameters (Δ(ΔH), Δ(ΔS)) of two adjacent benzodiazepines on a chromatogram were obtained from plots of lnα versus 1/T. We first studied each factor (fraction of water ϕ in the ACN/water mixture and column temperatureT), which controls the retention mechanism, and then we examined the simultaneous variation of all these factors. The changes in Δ(ΔH) and Δ(ΔS) in relation to a volume fraction of water ϕ in an ACN/water mixture were examined. In the ACN/water system, Δ(ΔH) was fairly constant in the acetonitrile region of ϕ≤0.52 and appears to be a roughly linear function of ϕ for ϕ≥0.52. In this system Δ(ΔS) is approximately a parabolic function of ϕ with an optimum at ϕ≅0.52. The retention mechanism of ten benzodiazepines was found to be significantly different in the methanol/water and ACN/water mixtures. The separation optimization of these ten benzodiazepines was then considered. A fraction of water of 0.43 in the ACN/water mixture and a column temperature of 44°C gave the most efficient separation conditions in the ACN/water mixture.
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  • 46
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polyurethane degradation products ; 2,4- and 2,6-toluendiamine ; Urine analysis ; Breast implants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A reversed-phase HPLC method has been developed for the urinary determination of mutagenic 2,4- and 2,6-toluendiamines. These amines are degradation products of polyurethane, a material used to cover textured silicone breast implants. FMOC-Cl was used as fluorescent derivatising agent in order to obtain a limit of detection of 15 ng/ml in urine. Pre-treatment of urine samples was by liquid/liquid extraction and urine specimens of patients after surgury were analysed.
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  • 47
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    Chromatographia 41 (1995), S. 657-660 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polyphenols ; Flavonoids ; Phenolic acids and aldehydes ; Eucalyptus spp.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Standards of the polyphenols occurring in wood, bark and leaf extracts ofEucalyptus spp. (i.e. flavonoids and phenolic acids and aldehydes) have been analyzed by HPLC using reversed phase columns, gradient elution and diode-array detection. The conditions used are reported.
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  • 48
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    Chromatographia 40 (1995), S. 657-661 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed phases ; Polyencapsulated silica ; Polyacrylates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary By copolymerization of silica modified with vinyl groups and acrylic acid esters in suspension highly efficient, polymeric-encapsulated stationary-phases can be prepared. The suspending liquid has to be selected such that the monomers are at least partially adsorbed on the surface and the polymer formed does not precipitate. Under these conditions stationary phases capable of speedy mass transfer can be prepared, where the reduced plate heights are between 2 and 3. The silica surface shielding is optimized so that basic and acidic solutes elute with symmetrical peak.
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  • 49
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chiral resolution of amino acids ; Pre-column derivatization ; Fluorescent chiral tagging reagents ; Stereochemical purity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Derivatization of amino acid enantiomers with fluorescent chiral Edman-type reagents, 4-(3-isothiocyanatopyrrolidin-1-yl)-7-nitro-2,1,3-benzoxadiazole [R(−)- and S(+)-NBD-PyNCS] and 4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N,N-dimethylaminosulfonyl)-2,1,3-benzoxadiazole [R(−)-and S(+)-DBD-PyNCS], yields corresponding diastereomers separable by reversed-phase HPLC on normal achiral columns. The resolved diastereomers were detected fluorometrically at 530 nm with excitation at 490 nm for the NBD-PyNCS derivatives and at 560 nm with excitation at 450 nm for those derived from DBD-PyNCS reagents. This HPLC-derivatization method was used for evaluation of stereochemical purity for some synthetic commercial peptides. The enantioanalysis was reliable down to 0.05% racemization of the amino acid residues and a quantity of 100 μg peptide sample was usually enough for the analysis. Two acid hydrolysis methods, i.e. the standard procedure with constant-boiling hydrochloric acid (HCl) and a rapid vapor-phase procedure with HCl-trifluoroacetic acid (TFA) mixture, were compared. The later was found to be unsuitable owing to increased racemization of the amino acid residues during the hydrolysis. Judging from the results obtained for proline and leucine residues, most of the tested peptides including biologically active peptides, such as neurotensin, [d-Ala2,d-Leu5]-enkepharin and morphine tolerance peptide, possessed stereochemical purities higher than 98%. Influence of structural features of the peptides on the racemization of the amino acid residues was found to be significant.
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  • 50
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Mass spectrometric detection ; Fast atom bombardment ionization ; Serum bile acid ; Rat bile
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary In order to analyse bile acids in biological matrices, e.g. rat bile and human serum, high performance liquid chromatography (HPLC) was coupled to continuous-flow fast atom bombardment mass spectrometry (CFFAB-MS). A gradient elution system which had already proved to be well suited for the quantitative determination of conjugated bile acids in bile was modified to allow HPLC-CFFAB-MS-coupling. Due to the sensitivity of this coupling method it is possible to obtain more information about the biliary bile acid pattern and species-specific secondary bile acids. Furthermore, we were able to identify obviously unknown bile acid species in rat bile which most likely classify as mono-oxo and di-oxo-taurocholates (MW 513 Da, 511 Da) and mono-oxo-glycocholates (MW 463 Da). In the present study we show that using this system it is possible to determine both conjugated and unconjugated as well as sulfated bile acids, without time consuming group separation and derivatization, from rat bile and human serum. In addition, it is suggested that the method presented here should be considered for use in routine analysis.
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  • 51
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fluorescence enhancement ; Bovine serum albumin ; Dansylamino acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The effects of acetonitrile and bovine serum albumin (BSA) concentrations on the signal intensity and retention behavior of dansylamino acids have been examined by using γ-cyclodextrin-bonded silica gel as the stationary phase in microcolumn liquid chromatography. Fluorescence intensities of dansylamino acids were enhanced by BSA as a mobile phase additive, e.g., detection limits of dansyl derivatives of L-Ala and L-Phe were improved by a factor of 12–18.
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  • 52
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Micellar liquid phases ; Octanol-water partition coefficient ; Benzene and naphthalene derivatives
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary It is shown that the logarithm of the capacity factor (log k′) for fifteen benzene and naphthalene derivatives in micellar liquid chromatography with forty nine different mobile phases generally correlates better with the logarithm of the octanol-water partition coefficient (log Pow) than the capacity factor (k′). Optimum conditions are established to obtain the best linear correlations of log k′-log Pow.
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  • 53
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Photolysis ; Electrochemical detection ; Drugs with chlorinated aromatic rings
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Pharmaceuticals containing a thiazide ring or chlorinated aromatic ring were investigated with respect to enhanced selectivity in determination. Oxidative electrochemical detection coupled with HPLC was used to study the influence of the pH of the mobile phase under conditions of photolysis. To cover a pH range 3.9–12, the employment of a polymer column stable in alkaline media was necessary. The method offers the great advantage of derivatization without chemicals at low operating potentials, thereby providing high selectivity.
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  • 54
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    Chromatographia 40 (1995), S. 296-302 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Structural isomer separation ; Cyclodextrin ; Polymers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Several β-cyclodextrin (CD)-bonded stationary phases for high performance liquid chromatography (HPLC) were prepared, based on silica beads coated with a poly(alkylamine), [poly(ethyleneimine)(PEI)]. In order to obtain stationary phases with a high content of CD and maximum accessibility of the CD cavity, the functionalization was carried out after the coating, using the mono-tosyl derivative of CD as the intermediate. The ability of these supports to separate ortho, meta and para isomers of some disubstituted benzene derivatives was examined. The contribution of the amino groups of the polymer and of the CD cavity to the separation process is discussed. The resolution is mainly based on the difference in the stability of the complexes of the various isomers with CD. The influence of the amount of CD on the height of the theoretical plate is also studied.
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  • 55
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed phase ; Polystyrene ; Pellicular ; Ultra high molecular mass
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Ultra high molecular mass (〉 7 million dalton) polystyrenes are prone to degradation in size exclusion chromatography. In gradient elution reversed phase HPLC they do not elute visibly on small particle size porous supports. However, large diameter C18 pellicular particles were successfully employed for reversed phase study of ultra high molecular mass (15 million dalton) polystyrenes without polymer degradation during elution. Although retention for the lower molecular mass polystyrenes was lower than on small diameter porous particles, the medium high molecular mass polystyrenes (0.5–1 million dalton) showed similar retention. The addition of small diameter porous particles in small quantities, to the large diameter pellicular particles, increased the amount of retention of the low molecular mass polystyrenes without affecting the higher molecular mass polystyrenes.
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  • 56
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Methyl ethyl ketone in urine ; Derivatization ; 3-Methyl-2-benzothiazolinone hydrazone ; Comparison with GC-MS
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A HPLC method for the determination of methyl ethyl ketone (MEK) in urine after derivatization with 3-methyl-2-benzothiazolinone hydrazone is proposed. The calibration curve for the ketone was linear, ranging between 0.23–10 mg/L, with a detection limit of 0.025 mg/L. The results were compared to those obtained by GC-MS, coupled to the headspace technique. MEK derivatization and the derivative purification processes were verified with respect to the main variables such as reaction temperature, reagent concentration, probable interferences and enrichment phase. The method is simple and reliable and shows a good sensitivity.
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  • 57
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    Chromatographia 40 (1995), S. 382-386 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Sulfonamides in foodstuffs ; Honey ; Milk ; Eggs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A procedure for the simultaneous determination of several sulfonamides in different foods, such as honey, milk and eggs is proposed. The analysis is carried out using reversed phase liquid chromatography with spectrophotometric detection. Optimization of the mobile phase led to good separation and a short analysis time when an initial isocratic step with a 3∶97 acetonitrile: water mixture was used for 5 minutes, followed by a linear gradient up to a 40∶60 mixture over 15 min. The proposed method is suitable for routine quality control analysis to ensure the absence of sulfonamides in foods. Recovery studies yielded good results for all food samples because there were no interferences from the matrices.
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  • 58
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    Chromatographia 40 (1995), S. 411-416 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Particle beam LC/MS interface ; Capillary columns for LC ; Mycotoxins ; Peanut meal
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A new method for the analysis of aflatoxins in food extracts, based on liquid chromatography/mass spectrometry interfacing, is presented. The chromatographic separation was performed with a reversed phase packed capillary column coupled with a modified particle beam interface capable of handling microliter per minute flow rates. This system allows higher overall sensitivity and easier operation procedures. The method has proved to be particularly suitable for the analysis of the toxins in very complex matrices. The specificity of electron impact ionization allowed positive identification of the aflatoxins with an excellent response linearity for accurate quantitation.
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  • 59
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Thin-layer chromatography ; Reversed phase ; 2-benzoylbenzoic acids ; Anionic and cationic counter ions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The 2-benzoylbenzoic acid series was investigated by reversed-phase, high-performance, thin-layer and column chromatography using various alkylammonium salts and di(2-ethylhexyl)orthophosphoric acid as polar associating reagents. The effects of the individual substituents on retention were quantified by Δlog k′ and ΔRM values. The compounds investigated differing in molecular structure (hydrophilic and hydrophobic substituents) commonly occurring groups in drugs and biologically active substances provide information on molecular interaction in these ion-pair systems. The combined effects on retention of organic modifier and ion-pair reagent concentration were investigated.
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  • 60
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Two-dimensional separation ; D- and L-amino acids ; Enantioseperation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A two-dimensional, column liquid chromatographic system is used for the determination of the D- and L-enantiomers of amino acids in biological samples. Separation of the amino acids is first on ion-exchange column by gradient elution with a sodium citratesodium chloride buffer. Enantioseparation is by subsequent injection of 3 μl heart-cuts of the individual amino acids onto a second column with a chiral crown ether stationary phase. Finally, fluorescence detection is after post-column labelling of the amino acids using ano-phthalaldehyde-2-mercaptoethanol reagent solution. The high separation power and selectivity of the system allow processing of complex samples with hardly any additional treatment and the determination of small quantities of D-amino acids in the presence of excess L-form. Applicability of the system is illustrated by the determination of D- and L-aspartate, serine, glutamate and alanine in various complex biological samples, such as protein hydrolysates, urine and biotechnological and food samples. Data are given on detectability, repeatability and linearity.
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  • 61
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion pair LC ; Sweeteners, preservatives and antioxidants ; Food additive analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A paired-ion, reversed-phase high-performance liquid chromatographic method was developed for the simultaneous determination of sweeteners (dulcin, saccharin-Na and acesulfame-K), preservatives (sodium dehydroacetate, sorbic acid, salicyclic acid, benzoic acid, succinic acid, methyl-para-hydroxybenzoic acid, ethylpara-hydroxybenzoic acid, n-propyl-para-hydroxybenzoic acid, isopropyl-para-hydroxybenzoic acid, n-butyl-para-hydroxybenzoic acid, and isobutyl-para-hydroxybenzoic acid), and antioxidants (3-tertiary-butyl-4-hydroxyanisole and tertiary-butyl-hydroquinone). A mobile phase of acetonitrile-50 mM aqueous α-hydroxy-isobutyric acid solution (pH 4.5) (2.2 ∶ 3.4 or 2.4 ∶ 3.6, v/v) containing 2.5 mM hexadecyltrimethylammonium bromide and a C18 column with a flow rate at 1.0 mL/min and detection at 233 nm were used. This method was found to be very reproducible with detection limits ranged from 0.15 to 3.00 μg. The retention factor (k) of each additive could be affected by concentrations of hexadecyltrimethylammonium bromide and α-hydroxyisobutyric acid, and pH and ratio of mobile phase. The presence of additives in some food samples was determined.
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  • 62
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Limit of detection ; Precision ; Uncertainty prediction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The precision of integration over noisy instrumental output for quantitative analysis is studied. A probability theory is developed to predict the relative standard deviation (RSD) of integration results over an integration domain from one-point integation (peak height measurement) to entire area integration in HPLC. Common integration modes of horizontal zero line and oblique zero line are taken into account, but no peak overlap is assumed. The question of the analytical superiority of peak height measurement or integration for quantitation is answered. In the HPLC apparatus used, the minimum RSD of measurements is found in the integration domain of ca. ±0.5 σ for analytes [peaks are approximated by the Gaussian signal of width, σ (standard deviation)]. The RSD of integration measurements is also shown to depend on the stochastic properties of back-ground noise (uncorrelated noise and correlated 1/f type noise). The theoretical conclusion is verified by Monte Carlo simulation and HPLC experiments for some aromatic compounds.
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  • 63
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion-pair and GPC separations ; Amoxicillin oligomers ; β-Lactam ring polymers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Polymeric substances formed from concentrated sodium amoxicillin in an aqueous solution have been separated using two high-performance liquid chromatographic methods. We used a C18 reversed-phase column with tetrabutylammonium chloride as an ion-pairing agent with an acetonitrile gradient and a TSKgel G2500PWxl column with water as the solvent for gel permeation chromatography. The separated materials, ranging in size from the monomer to the tetramer, have been characterized by functional-group chemical analysis, while the identification of the piperazine-2,5-dione was done using a pure standard. A greater number of peaks which were also better defined were obtained using the ion-pair reversed-phase method, and open and closed beta-lactam ring polymer forms could be distinguished. Using the gel permeation method, only a few monomer, piperazine-2,5-dione, dimer, trimer and combined amoxicillin trimer and tetramer peaks were obtained with water, although those obtained were quite well defined. The data on the time-course of formation of the oligomers and the amoxicillin degradation product were virtually identical by both methods.
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  • 64
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; On-column fluorimetric detection ; Gradient elution ; Dansyl amino acids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Fluorimetric detection in the presence of a stationary phase has been applied to gradient elution of dansyl amino acids in liquid chromatography. A 1.5 mm ID quartz tube packed with the same materials as the separation column was employed for the flow cell. Conventional-size columns were employed. The peak height of analytes increased with increasing retention owing to focusing and environmental effects of the stationary phase, leading to improvements in sensitivity, which was pronounced for analytes eluting late. The lower the gradient, the larger the improvement in sensitivity achieved. Detection limits were improved by a factor of up to 5.1 by fluorimetric detection using the packed flow cell, compared with those achieved using a common empty flow cell.
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  • 65
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; LC-MS ; Cinchona alkaloids ; New column packing materials
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A major problem in the HPLC analysis of alkaloids is the poor peak shape and consequently low resolution, due to the interactions of the basic alkaloids with the residual acidic silanol groups of most reversed phase materials. The performance of new packing materials specially designed for the separation of basic compounds has been studied using mobile phases without the special additives commonly applied in the analysis of alkaloids. Strongly basic Cinchona alkaloids were used as test compounds. Retention characteristics and selectivities of each material were studied, after mobile phase optimisation for the column. The influence of the major factors (nature and content of the organic modifier, pH value, salt concentration) affecting resolution was studied. The mobile phases were chosen so that they could be used in thermospray LC-MS. The addition of salts to the mobile phase improves separation but in general the modification of the mobile phase gave little change in selectivity. The performance of silicabased C18 material proved superior to the polymer materials tested.
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  • 66
    ISSN: 1573-5125
    Keywords: chlorophyll fluorescence ; nutrient limitation ; phytoplankton ; photosynthesis ; quantum efficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 67
    ISSN: 1573-5125
    Keywords: intertidal areas ; photosynthetically active radiation ; photosynthesis ; Tagus estuary ; tides
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photosynthetically active radiation (PAR) and temperature were measured continuously at the surface of estuarine intertidal sediments in the Tagus estuary, Portugal, along two spring-neap tidal cycles. PAR and temperature were strongly conditioned by the periodic tidal inundation, with large and abrupt variations occurring during flooding and ebbing. PAR levels reaching the sediment surface decreased very rapidly to zero or very low values during most of the daytime immersion. Inundation during high tide had the general effect of attenuating the amplitude of daily temperature fluctuation, with the incoming water usually warmer than the sediment during the night or early morning and cooler during the day. The daily progression of tidal emersion resulted in a clear fortnightly variation in total daily PAR reaching the sediment surface, while both daily mean temperature and mean temperature of diurnal low tide periods failed to exhibit a well-defined fortnightly periodicity. The obtained results indicate that the estuarine intertidal environment is dominated, at sub-seasonal time scales, by fortnightly periodicity in irradiance and temperature conditions favourable for benthic photosynthesis.
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  • 68
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    Journal of applied phycology 10 (1998), S. 547-554 
    ISSN: 1573-5176
    Keywords: herbicide ; green alga ; growth ; nutrients ; photosynthesis ; it Protosiphon botryoides ; respiration ; Thiobencarb
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of the herbicide thiobencarb (Saturn) were tested on the growth and physiology of the chlorophyte Protosiphon botryoides isolated from an Egyptian paddy. Assays were conducted using 16-day batch cultures. Chlorophyll and dry weight biomass yields were significantly reduced at 2–3 mg L-1 thiobencarb, and dark respiration increased and protein decreased significantly at 3 mg L-1. Reductions in exponential specific growth rate (μ) were generally small, but in some cases significant. Thiobencarb also slightly, but significantly, reduced the 77 K fluorescence parameter Fv/Fm, an indicator of maximum photosynthetic efficiency. No consistent dose-dependent changes occurred in chlorophyll per unit dry weight, total carbohydrate or gross photosynthetic capacity. Whereas half of the added thiobencarb was recovered from control (uninoculated) medium, it was largely absent from cells and culture medium after sixteen days, indicating biodegradation by the alga or associated bacteria. P. botryoides recovered fully within sixteen days following subculture in thiobencarb-free medium. Independently varying phosphate and nitrate nine-fold had no clear effect on the sensitivity of P. botryoides to thiobencarb.
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  • 69
    ISSN: 1573-5176
    Keywords: Cyanobacterium ; Spirulina platensis ; Arthrospira ; CO2 ; organic carbon ; nitrogen ; photosynthesis ; batch culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The consequences of the addition of CO2 (1%) in cultures of S. platensis are examined in terms of biomass yield, cell composition and external medium composition. CO2 enrichment was tested under nitrogen saturating and nitrogen limiting conditions. Increasing CO2 levels did not cause any change in maximum growth rate while it decreased maximum biomass yield. Protein and pigments were decreased and carbohydrate increased by high CO2, but the capability to store carbohydrates was saturated. C:N ratio remained unchanged while organic carbon released to the external medium was enhanced, suggesting that organic carbon release in S. platensis is an efficient mechanism for the maintenance of the metabolic integrity, balancing the cell C:N ratio in response to environmental CO2 changes. CO2 affected the pigment content: Phycocyanin, chlorophyll and carotenoids were reduced in around 50%, but the photosynthetic parameters were slightly changed. We propose that in S. platensis CO2 could act promoting degradation of pigments synthetised in excess in normal CO2 conditions, that are not necessary for light harvesting. Nitrogen assimilation was significantly not affected by CO2, and it is proposed that the inability to stimulate N assimilation by CO2 enrichment determined the lack of response in maximum growth rate.
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  • 70
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    Journal of applied phycology 11 (1999), S. 195-201 
    ISSN: 1573-5176
    Keywords: dense algal suspension ; light-harvesting pigment ; photosynthesis ; productivity ; cyanobacterium ; Synechocystis PCC 6714
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Microalgal productivity was examined using both a wild type and a phycocyanin-deficient mutant of Synechocystis PCC 6714 (PD-1). The culture was conducted at various light intensities under low and high cell densities in a continuous culture system. At low light intensity, photosynthetic productivity was almost the same for both low and high cell densities. However, at higher light intensities photosynthetic productivity was higher in mutant PD-1 than in the wild type. At 2000 μmol photon m−2 s−1 the productivity was 50% higher in mutant PD-1. This result is consistent with our first report (Nakajima & Ueda, 1997), which showed that photosynthetic productivity can be improved by reducing the light harvesting pigment content in high cell density cultures at high light intensities. It is concluded that the technology for reducing LHP content is a useful method for improving photosynthetic productivity in algal mass production.
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  • 71
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    Journal of applied phycology 8 (1996), S. 263-273 
    ISSN: 1573-5176
    Keywords: molecular biology ; mutagenesis ; photosynthesis ; protein engineering ; respiration ; thylakoid membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cyanobacteriumSynechocystis sp. PCC 6803 is readily amenable to targeted mutagenesis: Foreign DNA is taken up spontaneously, and after uptake DNA can be integrated into the organism's genome by homologous recombination. Using appropriate DNA constructs for transformation, specific genes in the organism can be interrupted, deleted, or replaced by modified gene copies. The organism can grow under a number of different conditions, ranging from photoautotrophic to fully heterotrophic modes, making genetic modifications that alter fundamental processes such as photosynthesis and/or respiration feasible. For example, deletion of photosystem I leads to an obligate (photo)heterotrophic strain in which photosystem II-generated electrons appear to be consumed by respiratory processes, whereas deletion of photosystem II leads to an obligate (photo)heterotrophic strain in which cyclic electron flow around photosystem I appears to remain active. A major advantage ofSynechocystis sp. PCC 6803 is that its entire genome has been sequenced (by S. Tabata and co-workers), opening many avenues to address basic and applied research problems. For example, genes can be introduced, modified or deleted, and hypotheses regarding the function of an open reading frame can be tested by deletion of this open reading frame. Methods to modify genes are numerous. In addition to site-directed mutagenesis, novel molecular genetic approaches including ‘targeted random mutagenesis’, combinatorial mutagenesis and introduction of hybrid genes have come of age and have proven to be very powerful tools in protein engineering. These approaches have been utilized primarily in this strain to study photosynthesis, but applications of this technology, including pathway engineering, alterations of substrate specificity of enzymes and introduction of tolerance to a variety of stresses, are equally feasible in relation to more applied aims. For optimal utilization of the potential of theSynechocystis sp. PCC 6803 system, however, an increased emphasis toward understanding the biochemistry and molecular physiology of cyanobacteria will also be critically important.
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  • 72
    ISSN: 1573-5176
    Keywords: fluorescence ; photoinhibition ; photosynthesis ; Spirulina ; photobioreactor ; temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A chlorophyll fluorescence technique was applied to anin situ study on the effects of low temperature and high light stresses onSpirulina cultures grown outdoors in controlled tubular photobioreactors at high (1.1 g L−1) and low (0.44 g L−1) biomass concentrations. Diurnal changes in PSII photochemistry (F v/F m) after 15 min of darkness, or in the light (dF/F′ m), and non-photochemical (qN) quenching were measured using a portable, pulse-amplitude-modulated fluorometer. The depression of theF v/F m ratio ofSpirulina cultures grown outdoors at 25°C (i.e. 10°C below optimum for growth) and 0.44 g L−1, reached 30% at the middle of the day. At the same time of the day thedF/F′ m ratio showed a reduction of up to 52%. The depression of bothF v/F m anddF/F′ m was lower in the cultures grown at 1.1 g L−1. Photoinhibition reduced the daily productivity of the culture grown at 0.44 g L−1 and 25°C by 33% with respect to that grown at 35°C. Changes in the growth yields of the cultures grown under different temperatures and growth rates correlate well with analogous changes in photon yield (dF/F′ m). Simple measurements of photochemical yield (F v/F m) can be used to test the physiological status ofSpirulina cultures. The results indicate that the saturating pulse fluorescence technique, when usedin situ, is a powerful tool for assessment of the photosynthetic characteristics of outdoor cultures ofSpirulina.
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  • 73
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    Journal of applied phycology 10 (1998), S. 419-425 
    ISSN: 1573-5176
    Keywords: Gracilaria cornea ; photosynthesis ; respiration ; chlorophyll ; phycoerythrin ; Florida ; salinity ; temperature ; irradiance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The agarophyte Gracilaria cornea, collected over 2.5 y in the Florida Keys, shows adaptations to oceanic salinities and subtropical to tropical water temperatures in its photosynthetic and respiratory responses as measured with a respirometer. No seasonal pattern in responses to irradiance, temperature, and salinity were evident between five collections over a 20-month period, indicating the tropical nature of the populations from Bahia Honda and Pigeon Keys. Concentrations of chlorophyll a (0.09 to 0.41 mg g d wt-1) and phycoerythrin (0.06 to 0.36 mg g d wt- 1) were low and reflect the low nutrient regime of the habitats, especially when compared to laboratory cultured plants. Compensation and saturation irradiances were also low (11–38 and 90–127 μmol photon m-2 s-1), indicating acclimation to lower irradiances in their shallow (1–2 m depth) habitats where turbidity can be high. In comparison with other subtropical and warm temperate species of Gracilaria, G. cornea had lower levels of pigment, but similarly high photosynthetic efficiency, demonstrating shade adaptation; it had only limited tolerance to salinities below 20‰ and temperatures below 15 °C. Thus, G. cornea from the Florida Keys in mariculture would require subtropical to tropical temperatures and stable oceanic salinities.
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  • 74
    ISSN: 1573-5176
    Keywords: Chlorophyll antenna size ; damage and repair cycle ; photon use efficiency ; photosynthesis ; photoinhibition ; Dunaliella salina
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The photon use efficiencies and maximal rates of photosynthesis in Dunaliella salina (Chlorophyta) cultures acclimated to different light intensities were investigated. Batch cultures were grown to the mid-exponential phase under continuous low-light (LL: 100 μmol photon m-2 s-1) or high-light (HL: 2000 μmol photon m-2 s-1) conditions. Under LL, cells were normally pigmented (deep green) containing ∼500 chlorophyll (Chl) molecules per photosystem II (PSII) unit and ∼250 Chl molecules per photosystem I (PSI). HL-grown cells were yellow-green, contained only 60 Chl per PSII and 100 Chl per PSI and showed signs of chronic photoinhibition, i.e., accumulation of photodamaged PSII reaction centers in the chloroplast thylakoids. In LL-grown cells, photosynthesis saturated at ∼200 μmol photon m-2 s-1 with a rate (Pmax) of ∼100 mmol O2 (mol Chl)-1 s-1. In HL-grown cells, photosynthesis saturated at much higher light intensities, i.e. ∼2500 μmol photon m-2 s-1, and exhibited a three-fold higher Pmax (∼300 mmol O2 (mol Chl)-1 s-1) than the normally pigmented LL-grown cells. Recovery of the HL-grown cells from photoinhibition, occurring prior to a light-harvesting Chl antenna size increase, enhanced Pmax to ∼675 mmol O2 (mol Chl)-1 s-1. Extrapolation of these results to outdoor mass culture conditions suggested that algal strains with small Chl antenna size could exhibit 2–3 times higher productivities than currently achieved with normally pigmented cells.
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  • 75
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    Journal of applied phycology 11 (1999), S. 157-163 
    ISSN: 1573-5176
    Keywords: Euglena gracilis ; photosynthesis ; waste water ; pulp and paper industry ; ultraviolet-B radiation (280–320 nm) ; pentachlorophenol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The green flagellate Euglena gracilis has been used as a model organism to elucidate the possible large-scale and short-term effects of waste substances from the pulp and paper industry on photosynthetic efficiency (PE). Different concentrations of waste substances before and after treatment in a cleaning system were studied. The uncleaned sample at concentrations up to 1:10 and the cleaned sample at concentrations up to 1:5 showed stimulating effects on the PE after 7 days of incubation compared to the control. The effects of waste substances on the PE of E. gracilis were also studied in combination with short-term studies (20 and 40 min) of ultraviolet-B radiation (UV-B, 280–320 nm). It was shown that increasing concentrations of the uncleaned sample had continuously stimulating effects on the PE and worked protectively against UV-B radiation. The cleaned sample exhibited no effects, or negative effects, on the PE of E. gracilis together with UV-B radiation compared to the experiments with only UV-B radiation. At the concentration 1:1 of the cleaned sample an increase in the PE was detected due to the high concentration of the coloured substances and a decrease in the UV-B penetration. PE revealed itself to be highly sensitive for detecting toxic effects on E. gracilis and is thus very promising for use in regular toxicity tests of waste water from pulp and paper industry.
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  • 76
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    Journal of applied phycology 10 (1998), S. 51-53 
    ISSN: 1573-5176
    Keywords: blue-green alga ; cyanobacterium ; Fv/Fmlight ; Nostoc flagelliforme ; photosynthesis ; rewetting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract PS II photochemical efficiency (Fv/Fm) of Nostoc flagelliforme was examined after rewetting in order to investigate the light-dependency of its photosynthetic recovery. Fv/Fm was not detected in the dark, but was immediately recognized in the light. Different levels of light irradiation (4, 40 and 400 µmol photon m2 s-1) displayed different effects on the recovery process of photosynthesis. The intermediate level led to the best recovery of photochemical efficiency; the low light required longer and the high light inhibited the extent of the recovered efficiency. It was concluded that the photosynthetic recovery of N. flagelliforme is both light-dependent and influenced by photon flux density.
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  • 77
    ISSN: 1573-5176
    Keywords: ammonium ; C:N ratio ; tank culture ; dietary fibre ; fatty acids ; nitrogen ; photosynthesis ; Ulva rigida
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Physiological and biochemical changes in relation to inorganic nitrogen availability were studied for tank-cultivated Ulva rigida grown under nitrogen- enriched and nitrogen-depleted seawater. U. rigida was initially cultivated in nitrogen-enriched seawater (daily concentrations of NH4+ and NO3- + NO2- ranged between 0.5–1.7 and 0.06–0.15 mg L-1, respectively), then transferred to nitrogen-depleted seawater where photosynthetic capacity decreased to zero after 23 d. At the time (14 d) when photosynthetic rates were lower than 2.0 μmol O2 g-1 FW min-1 and strong bleaching had occurred, some algae were returned to the initial nitrogen-enriched seawater to study recovery from N-limited growth. Data on biochemical composition (chlorophylls, ash, caloric content, fatty acids and dietary fibres) and colouration varied significantly depending on the nitrogen conditions. C:N ratios correlated significantly with biochemical parameters. Fatty acid (FA) synthesis continued during the N-starvation period; saturated and mono-unsaturated FA increased to a maximun of 72.2%, while poly-unsaturated fatty acids (PUFA) decreased to 27.7%. During the N-enriched recovery period, the reverse was found. C:N ratios above 10 correlated with carbohydrate synthesis as shown by the dietary fibre level. Under nitrogen enriched conditions, C:N ratios decreased along with a decrease in fibre level. Under controlled conditions, nitrogen represents a major influence on the development of intensive tank cultivation of Ulva rigida, not only by affecting parameters closely related to nitrogen metabolism but also some clearly influenced by carbon uptake.
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  • 78
    ISSN: 1573-5176
    Keywords: herbicide ; DCMU ; photosynthesis ; monoalgal culture ; Chaetoceros ; Dunaliella ; Nannochloropsis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The selective effect of DCMU on photosynthetic activity and growth rate was examined in several marine unicellular algae:Nannochloropsis sp. (Eustigmatohyceae),Dunaliella salina (Chlorophyceae)Isochrysis galbana (Prymnesiophyceae) andChaetoceros sp. (Bacillariophyceae). DCMU at 10−7 M caused an immediate decrease in the photosynthetic rate ofDunaliella andIsochrysis (about 50% inhibition), whereas 10−6 M imposed 80% inhibition in the photosynthetic rate ofChaetoceros. InNannochloropsis the rate was affected only when DCMU concentration exceeded 10−6M. Cellular growth rate of all studied algae was affected by DCMU in a similar manner to photosynthesis. The differential effect of DCMU was further examined in mixed cultures in whichNannochloropsis was cultivated together with an additional species simulating a contamination situation of aNannochloropsis culture. When DCMU was added at concentrations higher than 10−7 M, the growth of the competing algae significantly decreased, whileNannochloropsis maintained a relatively high growth rate. Consequently, after a growth period of 4 to 7 days a clear domination ofNannochloropsis was observed. These results demonstrate that DCMU and probably other herbicides of similar characteristics can be used effectively as a selective tool to suppress contaminating unicellular algae in open ponds in order to maintain a monoculture ofNannochloropsis.
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  • 79
    ISSN: 1573-5176
    Keywords: Spirulina platensis ; cyanobacterium ; NaCl ; photoinhibition ; photosynthesis ; photosystem II ; salt stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The response ofSpirulina platensis cells to salinity stress was studied. Once adapted to the higher osmoticum, photosynthetic parameters such as the maximum rate of photosynthesis under saturating irradiance (Pmax) and the initial slope of the P-I curve (α) are reduced by 15% and 25% in 0.5 M NaCl grown cells, respectively. Salt-adapted cells have a modified biochemical composition; reduced protein and chlorophyll content, and an increased level of carbohydrates. The reduction in the photosynthetic capacity of the salt-adaptedSpirulina cells reflects a lower ability to utilize light energy and results in an increase in the susceptibility of the stressed cells to photoinhibition. This conclusion is supported by the finding that cultures exposed to salt stress show not only a decrease in growth rate (μ), but lose the ability to respond to increased irradiance with an increase in growth. The use of variable fluorescence as a fast and reliable measurement to follow the changes in PSII of salt-stressesSpirulina cells enables following the early events of salinity shock. It indicates that as soon as the cells are exposed to salt, a protection mechanism is induced. This mechanism does not require any protein synthesis and may take place even in the dark, though at somewhat reduced effectiveness. The significance of the result in providing a better understanding of the interaction between two environmental stresses — light and salinity — and their application in the outdoor mass cultivation ofSpirulina are discussed.
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  • 80
    ISSN: 1573-5176
    Keywords: algal growth ; Porphyra ; inorganic carbon (Ci) ; photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photosynthetic (oxygen evolution) and growth (biomass increase) responses to ambient pH and inorganic carbon (Ci) supply were determined for Porphyralinearis grown in 0.5 L glass cylinders in the laboratory, or in 40 L fibreglass outdoor tanks with running seawater. While net photosynthetic rates were uniform at pH 6.0–8.0, dropping only at pH 8.7, growth rates were significantly affected by pH levels other than that of seawater (c. pH 8.3). In glass cylinders, weekly growth rates averaged 76% at external pH 8.0, 13% at pH 8.7 and 26% at pH 7.0. Photosynthetic O2 evolution on a daily basis(i.e. total O2 evolved during day time less total O2 consumed during night time) was similar to the growth responses at all experimental pH levels, apparently due to high dark respiration rates measured at acidic pH. Weekly growth rates averaged 53% in algae grown in fibreglass tanks aerated with regular air (360 mg L-1 CO2) and 28% in algae grown in tanks aerated with CO2-enriched air (750 mg L-1 CO2). The pH of the seawater medium in which P. linear is was grown increased slightly during the day and only rarely reached 9.0. The pH at the boundary layer of algae submerged in seawater increased in response to light reaching, about pH 8.9 within minutes, or remained unchanged for algae submerged in a CO2-free artificial sea water medium. Photosynthesis of P. linearissaturated at Ci concentrations of seawater (K0.5560 μM at pH 8.2) and showed low photosynthetic affinity for CO2(K0.5 61 μM) at pH 6.0. It is therefore concluded that P. linearisuses primarily CO2 with HCO3 - being an alternative source of Ci for photosynthesis. Its fast growth could be related to the enzyme carbonic anhydrase whose activity was detected intra- and extracellularly.
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  • 81
    ISSN: 1573-5176
    Keywords: CO2 ; inorganic carbon ; macroalgae ; photosynthesis ; PAM
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photosynthesis and cell composition of Porphyraleucosticta discs grown at low (〈 0.0001% in air), current (control) and high (1% CO2 in air)inorganic carbon (Ci) concentrations were analyzed. Carbohydrate content in discs grown at high Ci increased (15.1 mg g-1 FW) with respect to the control (6.4 mg g FW-1), whereas soluble protein content decreased to one-third (5.6 to2.1 mg g-1 FW). Carbohydrate content was unaffected and soluble protein slightly increased in discs grown at low Ci. As a consequence of these changes, a lower C/N molar ratio (8.6) was found in the discs grown at low compared to high Ci(12.4). Nitrate reductase activity increased at high Ci from 0.3 ± 0.2 to 1.7 ± 0.4 μmolNO2 - g-1 FW h-1indicating that reduction and assimilation of nitrate were uncoupled. The response of photosynthesis to increasing irradiance, estimated from O2evolution vs. irradiance curves, was affected by the treatments. Maximum quantum yield (Φ O2°) and effective quantum yield (Φ O2) at 150 μmol photon m-2s-1 decreased by 20% and 50%, respectively, at low Ci. These differences could be due to changes in photosynthetic electron flow between PSII and PSI. Treatments also produced changes in maximal (Fv/Fm) and effective (ΔF/Fm′)quantum yield for photosystem II charge separation.
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  • 82
    ISSN: 1573-5176
    Keywords: cell-wall polysaccharides ; chlorophylla ; Gelidium sesquipedale ; internal C and N ; light quality ; photosynthesis ; phycobiliproteins ; respiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The influence of different light qualities on the photosynthetic rate, dark respiration, intracellular carbon and nitrogen content, and accumulation of photosynthetic pigments and cell-wall polysaccharides during short-term incubation (5 h) of the red algaGelidium sesquipedale was investigated. The same photon irradiance of 50μmol m−2 s−2 below the light saturation point of photosynthesis was applied in each case. Blue light stimulated photosynthesis, dark respiration and the accumulation of chlorophyll and biliproteins, phycoerythrin in particular. The accumulation of internal carbon and nitrogen was greater under blue light than under the other light qualities. In contrast, the percentage of cell-wall polysaccharides was higher in red light. The content of cell-wall polysaccharides decreased during the time of incubation in all light treatments except in red light. The action of a non-photosynthetic photoreceptor in the control of cell-wall polysaccharide synthesis is suggested because the accumulation of cell-wall polysaccharides was not correlated with net photosynthesis in contrast to what occurred with carbon, chlorophyll and phycoerythrin accumulation.
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  • 83
    ISSN: 1573-5176
    Keywords: cyanobacteria ; light intensity ; photoinhibition ; photosynthesis ; recovery ; Spirulina platensis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three isolates ofSpirulina platensis (Norst) Geitler marked BP, P4P and Z19/2 were compared with respect to their response and acclimation capability to high photon flux densities (HPFD). Cultures exposed to HPFD (1500–3500 μmol photon m−2 s−1) exhibited a marked decrease in light-dependent O2 evolution rate. P4P was more sensitive to HPFD than the two other isolates. All three isolates recovered from photoinhibition when placed under low PFD. The BP isolate was able to recover also in the dark but to a lower extent and at a lower rate, while no recovery was observed in the other two isolates under dark conditions. No recovery was observed when protein synthesis was inhibited using chloramphenicol. Cultures grown at 200 μmol photon m−2 s−1 differed from cultures grown at 120 μmol photon m 2 s-1 by their lower maximal photosynthetic rate (P max ) and higher light saturation (I k ) value, while being more resistant to HPFD stress. The ability ofSpirulina isolates to acclimate and withstand HPFD may provide useful information for the selection of strains useful for outdoor mass cultivation.
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  • 84
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    Journal of applied phycology 9 (1997), S. 525-532 
    ISSN: 1573-5176
    Keywords: Gelidiella ; agarophyte ; photosynthesis ; P-I curve ; morphology ; tide ; pigment
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The red seaweed agarophyte, Gelidiella acerosa (Forsskål) (Feldmann& Hamel) was collected from tidepools, high intertidal rocks. and shallow subtidal are as along a reef flat in Ilocos Norte, northern Philippines. The three populations were compared during the summer (dry) and rainy (wet) seasons to determine changes in morphology and photoacclimation capacity as possible use in mariculture. During summer months (February toApril) after exposure to environmental extremes (i.e. the highest percent of minus tides during daylight, high light regimes, desiccation, and solar bleaching), the populations differed in their morphologies and responses to increasing irradiance levels (P–I curve). Tidepool plants were the tallest, bushiest, and with increased diameter of cortical cells; while,high intertidal plants were the shortest, with sparse branching pattern and decreased diameter of cortical cells. Although their saturation irradiances indicated shade tolerance (Ik = 52 − 112 µmol photon m -2 s-1). their differential light saturation curves (P-I curves) suggested a capacity to acclimate to ambient light regimes. For example, plants from the high intertidal zone showed higher photosynthetic rates and saturation irradiances, slightly lower initial slopes of the P-I curves and levels of light harvesting accessory pigments, rphycoerhythrin (R-PE) and rphycocyanin (R-PC), after being exposed to higher light regimes. In contrast, plants from tidepools and shallow subtidal areas had lower photosynthetic rates and saturation irradiances, slightly steeper initial slopes of the P-I curves and levels of R-PE and R-PC, having been exposed to lower light regimes. During the rainy months (June to November) no significant responses in these parameters were recorded. Comparison of the P-I responses of vegetative and tetrasporic plants showed these to vary with season. The data suggest that when plants became reproductive their physiological fitness either was unchanged or slightly enhanced. These results indicate that all three populations of G. acerosa could be used as seed stock for mariculture.
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  • 85
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    Journal of applied phycology 9 (1997), S. 503-510 
    ISSN: 1573-5176
    Keywords: Chlorella pyrenoidosa ; dense algalsuspension ; light-harvesting pigment ; photosynthesis ; Synechocystis PCC 6714
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of light-harvesting pigments (LHP) inmicroalgal cells on photosynthetic activity in adense cell suspension were examined. The results suggest that a lower LHP content should result in higher photosynthetic productivity under high light intensity. The idea was first proposed by Lien and San Pietro in 1975 that photosynthesis could be improved by reducing the LHP content in microalgal cells, but this has not been demonstrated in detail. Experiments to evaluate the idea were conducted with Synechocystis PCC6714 and Chlorellapyrenoidosa. In the experiments with PCC 6714, photosynthesis of a phycocyanin-deficient mutant was compared with that of the wild type. In the experiments with C. pyrenoidosa, the LHP content was controlled by the light intensity in the algalculture. The maximum photosynthetic activity was 20–30% higher in the dense suspension of cells having a lower LHP content with both organisms. These results indicate that the idea of reducing the LHP contentcould be applicable to a wide variety of photosynthetic organisms.
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  • 86
    ISSN: 1573-5176
    Keywords: Gelidium sesquipedale ; photosynthesis ; fluorescence ; light response curves ; pigments ; depth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photosynthesis-light response curves of Gelidium sesquipedale from the west coast of Portugal (Cape Espichel) were determined at four different depths, 3, 10, 15 and 22 m. Data acquisition using chlorophyll a fluorescence methodology and oxygen electrode measurements were compared. Response curves were determined over an increasing range of irradiance values (I), from darkness to 900 μmol photon m-2 s-1 PAR. In general, light response curves obtained for G. sesquipedale showed a similar pattern whether determined by the chlorophyll fluorescence method or by oxygen evolution. The photosynthetic capacity of G. sesquipedale decreased with depth, as expected, revealing a ‘sun’ and ‘shade’ acclimation pattern, between shallow and deeper waters.
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  • 87
    ISSN: 1573-5176
    Keywords: UV-radiation ; chlorophyll fluorescence ; photosynthesis ; stress tolerance ; electron transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Photoinhibition and recovery kinetics after short exposure to solar radiation following three different irradiance treatments of irradiances (PAR, PAR+UVA and PAR+UVA+UVB) was assessed in two intertidal species of the genus Gelidium, Gelidium sesquipedale and G. latifolium, collected from Tarifa (southern Spain) using in vivo chlorophyll fluorescence (PAM fluorometry). After 3 h UV radiation exposure, optimal quantum efficiency (Fv/Fm) in G. sesquipedale decreased between 25 and 35% relative to the control. Under PAR alone, values decreased to 60%. In G. latifolium, photoinhibition did not exceed 40%. Similar results were found for the effective quantum yield (ΔF/Fm′), however, no marked differences in relation to light treatments were seen. When plants were shaded for recovery from stress, only in G. latifolium a significant increase in photosynthesis was observed (between 80 and 100% of control). In contrast, photosynthesis of G. sesquipedale suffered a chronic photoinhibition or photodamage under the three light irradiances. Full solar radiation (PAR+UVA+UVB) affected also the electron transport rate in both species. Here, initial slopes of electron transport vs. irradiance curves decreased up to 60% of controls. Although the recovery kinetic under PAR+UVA+UVB conditions was delayed in G. latifolium, after 24 h recovery this species reached significantly higher than G. sesquipedale. PAR impaired electron trasport only in G. sesquipedale. Overall, both species are characterized by different capacity to tolerate enhanced solar radiation. G. latifolium is a sun adapted plant, well suited to intertidal light conditions, whereas G. sesquipedale, growing at shaded sites in the intertidal zone, is more vulnerable to enhanced UV radiation.
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  • 88
    ISSN: 1573-5176
    Keywords: 14C ; photosynthesis ; population growth ; Selenastrum capricornutum ; suspended sediment elutriate ; zinc ; cadmium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Short-term 14C-fixation (4 h) Selenastrum capricornutum algal toxicity tests were conducted with Cd (n=8), Zn (n=9) and suspended sediment aqueous elutriates (n=28) and the results were compared to those obtained in a 48 h population growth test. In order to provide more realistic experimental conditions, toxicity tests were carried out in prefiltered nutrient-spiked Lake Geneva water. The population growth inhibition test was significantly more sensitive than the14 C-fixation test for Cd (median EC50-4h and EC50-48h values of 600 and 118 µg L-1, respectively) whereas no significant difference was measured for Zn toxicity (median EC50-4h and EC50-48h values of 97 and 96 µg L-1, respectively). With suspended sediment aqueous elutriates, the relative sensitivity of the two different end points is sample dependent, with ratios of the EC25 for the14 C-fixation: population growth test ranging from 〈0.26 to 〉53.3. Elutriate toxicity shows no apparent relationship between the acute and chronic test, indicating that population growth inhibition cannot be derived directly or predicted from14 C-fixation. Both tests with their specific advantages and limitations provide valuable complementary information to measure the impact of single toxicants or complex mixtures on aquatic plants.
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  • 89
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    Journal of applied phycology 10 (1998), S. 447-452 
    ISSN: 1573-5176
    Keywords: dense algal suspension ; light-harvesting pigment ; photosynthesis ; Synechocystis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects on photoinhibition of light-harvesting pigments in microalgal cells were examined using the wild type and a phycocyanin- deficient mutant (PD-1) of Synechosystis PCC 6714. Mutant PD-1 showed higher resistance to high light than the wild type in terms of the decline of photosynthetic activity at any light intensity and with various cell densities. This suggests that the loss of productivity induced by high light intensity would be improved by reducing the content of light-harvesting pigments.
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  • 90
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    Journal of applied phycology 11 (1999), S. 473-477 
    ISSN: 1573-5176
    Keywords: aquaculture ; light ; photosynthesis ; Porphyra
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Due to their rapid growth and nutrient assimilation,Porphyra spp. are good candidates for bioremediation and polyculture. The production potential of two strains of P. purpurea and P. umbilicalis from north-east USA was evaluated by measuring rates of photosynthesis (as O2evolution) of material grown at 20 °C. Photosynthetic rates of P. umbilicalis were 80%higher than P. purpurea over the temperature range 5–20 °C, at both sub-saturating andsaturating irradiances (37 and 289 μmol photonm-2 s-1). Porphyra umbilicalis was more efficient at low irradiances (higher α) and had a higher Pmax (23.0 vs 15.6 μmolO2 g-1 DW min-1) than P.purpurea, suggesting that P. umbilicalis is a better choice for mass culture, where self-shading maybe severe.
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  • 91
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    Journal of applied phycology 11 (1999), S. 535-541 
    ISSN: 1573-5176
    Keywords: cyanobacterium ; Nostoc flagelliforme ; nutrients ; photosynthesis ; potassium ; re-hydration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of nutrients on the photosynthetic recovery of Nostoc flagelliforme during re-hydration were investigated in order to see if their addition was necessary. Net photosynthesis was negligible in distilled water without nutrient-enrichment. Addition of K+ resulted in significant enhancement of net photosynthesis, whereas other nutrients (Fe3+, Mg2+, Na+, NO3 -, PO4 3-, Cl-) and trace-metals (A5) showed little effect. The recovered net photosynthetic activity increased with the increased K+, and reached the maximum at concentrations above 230 μM. Desiccation and re-hydration did not affect the dependence of photosynthetic recovery on K+. It was concluded that dried field populations of N. flagelliforme require exogenous addition of potassium for photosynthetic recovery and that growth may be potassium-limited in nature.
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  • 92
    ISSN: 1573-5125
    Keywords: sherbicide tolerance ; photosynthesis ; phytoplankton ; simetryn ; triazine ; herbicides ; ultraviolet radiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We examined the effects of UV B radiation on14C-uptake rates and carbon assimilation into the major end-products of photosynthesis of the green algaScenedesmus in the presence and absence of the triazine herbicide simetryn. Experiments were conducted using both a herbicide-susceptible and herbicide-tolerant strains ofScenedesmus. Three different UV-B dose rates were used as well as a light control. The lowest dose rate was almost the same level as in subsurface of ponds and lakes, while the other two were slightly lower and higher than natural sunlight on the surface of ponds and lakes, respectively. Total uptake rates of14C were not reduced by the UV B irradiation alone even at the highest dose rate. However, in the presence of the herbicide, uptake rates were clearly reduced by the highest dose rate of UV-B concomitant with increasing herbicide concentrations in the herbicide-susceptible strain. On the other hand, the proportion of lipid fraction was slightly reduced by all the UV-B treatments in the herbicide-susceptible strain even in the absence of the herbicide. In the herbicide-tolerant strain, uptake rates were not affected by UV-B radiation or by the herbicide. These facts indicated that UV-B effects could be smaller than predicted. It may be important to examine combined effects of UV-B and other anthropogenic and/or natural stresses for assessing actual UV-B effects in the field.
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  • 93
    ISSN: 1573-5125
    Keywords: cryptomonads ; macromolecular ; Phototron ; photosynthesis ; UV radiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We used a device called a Phototron to measure the effects of UV radiation on the cosmopolitan algae, Cryptomonas erosa, grown in continuous cultures. In the Phototron, we investigated changes in photosynthetic parameters (Pmax – specific production rate at optimal light intensity; α – initial slope of the linear portion of the Photosynthesis-Irradiance curve; and θ – the convexity or rate of bending) and carbon allocation as a function of irradiance at three different environmentally-realistic doses of UV radiation in unconditioned (no prior UV exposure) and conditioned algae (15 d previous UV exposure). For unconditioned control algae, Pmax-Total was lower, although not significantly, than the two highest UV treatments. For conditioned control algae, Pmax-Total was higher, although not significantly, than all UV treatments. Our data suggest that short term (4 h) exposure to low levels of UV (8.09 W m−2 unweighted) does not affect Pmax-Total in C. erosa, but does change the proportion of carbon allocated to lipids and proteins. Also, comparisons of lipids, polysaccharides and proteins as a percent of total carbon uptake between unconditioned and conditioned algae indicate that exposure history to UV radiation can have a negative impact on carbon allocation to lipids and proteins, in a wetland alga species that is crucial to the efficient transfer of energy through freshwater food webs.
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  • 94
    ISSN: 1573-868X
    Keywords: Carbon dioxide ; coral reefs ; atoll ; lagoon ; photosynthesis ; calcification ; total alkalinity ; partial pressure ; topography ; residence time
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences
    Notes: Abstract Factors controlling the CO2 system parameters, including the partial pressure of CO2 (PCO2) in coral reef waters, were investigated in three mid-oceanic reefs of the Indo-Pacific region. Surface water PCO2 in the lagoons of Majuro Atoll and Palau barrier reef in the Pacific were 25 µatm and 48 µatm higher than those of the offshore waters, respectively, while South Male Atoll lagoon of the Maldives in the Indian Ocean exhibited relatively small difference in PCO2 compared to the offshore water. Observations from Majuro Atoll and Palau barrier reef are consistent with the view that calcium carbonate production predominates in coral reefs. On the other hand, results from South Male Atoll can be attributed to the thorough flushing of the lagoon, which is connected to the open ocean by numerous deep channels. The offshore-lagoon PCO2 difference depends on system-level net organic-to-inorganic carbon production ratio while reef topography, especially residence time of the lagoon, has a secondary effect on the magnitude of the offshore-lagoon difference. A potential for releasing CO2 might be more evident in an enclosed atoll where the reef water has a longer residence time. Oceanic atoll and barrier reef lagoons, which are in the terminal stage of evolutionary history of oceanic volcanic islands, have the potential to release CO2 to the atmosphere.
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  • 95
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    Aquaculture international 4 (1996), S. 237-252 
    ISSN: 1573-143X
    Keywords: Calcification ; Environmental disturbances ; Eumelanin ; Great scallop (Pecten maximus) ; Shell colour ; microstructure and melanization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Modifications of great scallop (Pecten moximus) shells have been noticed in many sites of scallop fisheries in Brittany, especially in shallow waters. These calcification abnormalities are linked to the appearance of a brown colouration of the internal calcified shell layer, due to the presence of a eumelanin associated with the insoluble organic matrix of the biocrystals. The appearance of this pigment generates many disturbances of the calcified foliated microstructure of the scallop internal shell layer. The mantle structure is not modified in brown shells as compared with white ones. No pathogenic signs such as hyperplasia or haemocytic infiltration have been observed. According to this observation, we hypothesize that the brown colour phenomenon is more a result of environmental disturbance rather than a symptom of a pathogenic disease. The colour abnormalities of the internal shell layer can be detected by a spectral analysis of its reflectance before it can be detected with the naked eye. This method, correlated to microstructural observations, gives a rapid and precise analysis of the appearance of the pigmentation on adult or juvenile scallops. It may be a useful method for the evaluation of the influence of environmental parameters, for example, on calcification and its abnormalities.
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  • 96
    ISSN: 1573-5117
    Keywords: gas exchange ; mangrove ; photosynthesis ; salinity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Continuous measurements of gas exchange characteristics were made on two to nine year old hydroponically grown Avicennia germinans (L.) Stearn, Aegialitis annulata R. Br. and Aegiceras corniculatum (L.) Blanco maintained at 50 or 500 mol m−3 NaCl. In Avicennia germinans and Aegialitis annulata, CO2 assimilation rates were initially higher at 500 mol m−3 NaCl and decreased gradually towards the end of the photoperiod when rates were similar to those at the lower salinity. In Aegiceras corniculatum, assimilation rates were higher at 50 mol m−3 NaCl and about 55% lower at the higher salinity. In all three species, leaf conductance and transpiration exhibited trends similar to those for CO2 assimilation. Intercellular CO2 concentrations were similar at both salinities in Avicennia germinans and Aegialitis annulata, but considerably higher at the lower salinity in Aegiceras corniculatum. Water use efficiencies (WUE), although similar between salinity treatments in Avicennia germinans and Aegialitis annulata, were greater at the higher salinity in Aegiceras corniculatum. Data obtained from CO2 response curves indicated that assimilation at high salinity in Aegiceras corniculatum was limited by conductance, and to a lesser extent, by photosynthetic capacity. In Avicennia germinans and Aegialitis annulata, assimilation was greater at the higher salinity as indicated by increase in both the initial slope and the upper plateau of the CO2 response data. Greater assimilation at high salinity in Avicennia germinans and Aegialitis annulata may be attributed to lower carbon losses via photorespiration and to efficient salt excretion and sequestration.
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  • 97
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    Hydrobiologia 364 (1997), S. 199-208 
    ISSN: 1573-5117
    Keywords: Phytoplankton ; photosynthesis ; light ; temperature ; tidal freshwater ; irradiance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The photosynthetic response to irradiance wasquantified for phytoplankton from the tidalfreshwater Potomac River biweekly to monthly over aperiod of six years. Samples were collected from twoshallow embayments and portions of the deeper rivermainstem. Photosynthetic rate was measured in thelaboratory at in situ temperature over a range ofirradiance levels and photosynthetic parameters werecalculated using nonlinear regression.PB max,the maximum photosynthetic ratestandardized to chlorophyll a, increased withtemperature up to 25 °C with a Q10 of 2.02. Above 25 °C, PB max was essentiallyconstant with temperature. Lesser correlationbetween PB max and ambient irradiance couldbe explained by the correlation of irradiance withtemperature. α, the slope of the P–I curve atlow light, was correlated with both ambientirradiance and temperature. Highest α valueswere found in late summer when high temperature andintermediate ambient irradiance were observed. Spring and early summer were characterized by lowα. Despite low light penetration, Ik andα values were indicative of sun limitationpossibly due to intermittent high light levelsexperienced during mixing. Ik showed a clearseasonal trend directly related to days from summersolstice. Spatial patterns were minimal except thatIk was consistently lower in one shallowembayment than in the other two areas. Seasonalpatterns in photosynthetic parameters correspondedroughly to changes from a spring diatom populationto summer cyanobacterial assemblage.
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  • 98
    ISSN: 1573-5117
    Keywords: photon flux density ; intracellular metabolic pools ; proteins ; carbohydrates ; lipids ; polysaccharides ; photosynthesis ; phytoplankton
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of photon flux density (PFD) on the partitioning of photosynthetically fixed 14CO2-C into major intracellular end products was investigated for three species of freshwater planktonic algae (Nitzschia palea, Monoraphidium minutum and Synechococcus elongatus belonging to three different classes. This study was designed to investigate the phenomenon of polysaccharide synthesis associated with the saturation of protein synthesis and to test if this process is common to all three phytoplankton species. Protein synthesis was saturated at low PFD in all three species of algae studied. However, fixed carbon was differentially stored, namely in lipids in Nitzschia palea (Bacillariophyceae), in polysaccharides in Monoraphidium minutum (Chlorophyceae), and in low molecular weight metabolites (LMW) in Synechococcus elongatus (Cyanophyceae). The results of this transient state study indicate that the metabolic pathways of algae can easily be controlled by different irradiance. Furthermore, it appears that the difference in the patterns of synthesis is taxonomy dependent.
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  • 99
    ISSN: 1573-5117
    Keywords: biotest ; brackish ; Gracilaria ; growth inhibition ; marine ; photosynthesis ; Rhodophyta ; seaweed ; toxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A growth inhibition test method was developed using the macroalga Gracilaria tenuistipitata as the test organism. This alga was chosen because of its high laboratory growth rates, commonly 30–40% d−1, which are reached in salinities between 5 and 40‰, and its epiphyte resistance. The toxicity of a number of substances, including heavy metals, herbicides and complex wastewaters towards the alga was assayed. Anti-fouling paints were tested with a modification of the method. EC50 values for heavy metals varied between 0.05 and 17 mg l−1 and for herbicides between 0.002 and 0.02 mg l−l. The sensitivity to the toxicant was generally higher at low salinity. Omitting nitrogen and phosphorus additions to the test medium increased the sensitivity and a semi-static performance was possible with maintained or increased sensitivity. Preliminary tests done with a computerised photosynthesis inhibition method produced promising results. In conclusion, this is a simple, sensitive and reproducible test method for assessing the toxicity of substances, wastewaters and anti-fouling paints in brackish and marine environments.
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  • 100
    ISSN: 1573-5117
    Keywords: Avicennia marina ; gas exchange ; mangroves ; photosynthesis ; waterlogging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This study was undertaken in summer on fully expanded leaves of Avicennia marina trees in the Beachwood Mangroves Nature Reserve, Durban,South Africa. Data sets were obtained over 5–7 days of relatively dry conditions and over two periods of 5 days during which the swamp was continuously inundated with dilute seawater (〈 150 mol m−3NaCl). Gas exchange responses were strongly influenced by photosynthetic photon flux density (PPFD), leaf temperature and leaf to air vapour pressure deficit (Δw). Carbon dioxide exchange was saturated at a PPFD of about 800 µmol m−2 s−1. Maximal CO2 exchange rates ranged from 8.5 to 9.9 µmol m−2 s−1 with no differences between drained and waterlogged conditions. Under drained conditions, leaf conductance,transpiration and internal CO2 concentrations were generally lower, and water use efficiencies higher, than during waterlogging. Continuous waterlogging for 5 days had no adverse effect on CO2 exchange. Xylem water potentials ranged from −1.32to −3.53 MPa during drained and from −1.02 to −2.65 Mpa during waterlogged conditions. These results are discussed in relation to anatomical and metabolic adaptations of A. marina to waterlogging stress.
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