THE KINETICS OF THE EARLY STAGES OF THE PHYTOCHROME PHOTOTRANSFORMATION Pr→ Pfr. A COMPARATIVE STUDY OF SMALL (60 kDalton) and NATIVE (124 kDalton) PHYTOCHROMES FROM OAT

A comparative study of the decay kinetics of photogenerated transients from small (60 kDalton) and native (124 kDalton) oat phytochrome in the red-absorbing form (P_r) in phosphate buffer containing 5 mM ethylenediamine tetraacetic acid, pH 7.8, (PB) and in PB containing 20% ethylene glycol, has been carried out in the temperature range 275–298 K. The analysis confirmed that at least two primary photoproducts, intermediates Iⁱ₇₀₀s and Iⁱ_7oo are formed from P_r. The kinetic parameters, as observed in PB at 695 nm and 275 K, are similar for the I₇₀₀ intermediates of both small and native phytochrome. Namely, the lifetimes are about 21 μs (component percentages 38%) for the I Iⁱ₇₀₀s and about 200 μ.s (62%) for the Iⁱ₇₀₀S- Arrhenius preexponential factors (A) of about 10¹⁶ and 10¹⁵ s-¹and activation energies of about 61 and 56 kJ/mol were measured for the absorbance decays of the I₇₀₀S of small and native phytochrome, respectively. The kinetic data favour parallel paths for the formation of the Iⁱ₇₀₀s from P_r, and the activation parameters indicate that the primary photoreactions of the transformation from P_r to the far-red-absorbing form are restricted to the chromophore within the protein. Moreover, the relatively modest temperature dependence of the lifetimes of the Iⁱ₇₀₀^S from small and native P_r supports the working hypothesis that the ground state reactions to the Iⁱ_bl, intermediates–although somewhat influenced by the polypeptide fragment that is removed upon degradation of native to small P_r–are localized to the chromophore, as is most probably the case also for the primary photoreactions. The effect of the addition of 20% ethylene glycol on the pre-exponential factors of the time-dependent decay functions is discussed in similar terms of the early stages of the phototransformation.     

Synthesis and Monoamine Oxidase Inhibitory Activity of 3-Substituted 5H-Indeno[1,2-c]pyridazines

The one-pot synthesis of nine 5H-indeno[1,2-c]pyridazines is described. These compounds are shown to be potent, reversible inhibitors of monoamine oxidase B (MAO-B) with little or no effect on monoamine oxidase A (MAO-A). Qualitative structure-activity relations indicate that the MAO-B inhibitory activity is strongly influenced by electronic and bulk properties of substitutents.     

Enhancement of the oxygen transfer in a circulating three-phase fluidized bed bioreactor

Applied Biochemistry and Biotechnology - The addition of α-alumina to the aqueous solution of sodium alginate for imobilization of viable cells allows the production of denser particles than...     

Solution Conformations,Photophysics, and Photochemistry of Bile Pigments; Bilirubin and Biliverdin,Dimethyl Esters and Related Linear Tetrapyrroles

Bilirubin and biliverdin dimethyl esters (BRE and BVE, respectively) and related linear tetrapyrroles have been studied using a combination of photochemical and spectroscopic techniques, the latter including absorption, fluorescence fluorescence excitation, medium-induced circular dichroism, and proton magnetic resonance. Both types of tetrapyrroles form mixtures of different topological isomers in very dilute solutions. In the case of the bilirubins the heterogeneity of the solutions is caused by two coexisting conformers with different orientations of the A/B and C/D pyrromethenone moieties with repect to each other. The spectral properties of one conformer resemble the isolated parent pyrromethenone, whereas those of the other result from electronic coupling of the two subchromophores presumably held in a “ridge tile” -like orientation. C? C rotations at the C-5 and C-15 bridges substantially compete in both components with the photochemical channels (E→Z isomerization and lumirubin formation) for the radiationless deactivation of the excited singlet state. The more rigid “ridge tile” component additionally undergoes hydrogen bond-mediated deactivation, and it photoisomerizes more efficiently. The situation is markedly more complex with the biliverdins. In order to obtain a more detailed insight into the mechanisms of the radiationless excited-state processes, time-resolved optoacoustic spectroscopy and ultrafast absorption (pump-probe) and fluorescence detection (single-photon-timing) techniques were used to supplement the stationary methods. The solution mixtures are composed of a (family of) helically coiled all-Z, all-syn species, and of species differing from the former by stretched arrangements of the rings B and C around the central C-10 bridge (E-anti, E-syn, and Z-anti). Two excited singlet states with picosecond lifetimes are attributed to either one or two coiled ground-state forms, and two remarkably long-lived nanosecond excited states arise each from a stretched ground state. The radiationless deactivation of the shorter-lived of the picosecond states is brought about by ultrafast intramolecular proton transfer between the B/C nitrogen atoms, in addition to the C? C rotational modes operative in both. Z→E photoisomerization is also an appreciable deactivation channel of excited biliverdin dimethyl ester. It is confined to the central C-10 double bond and selectively affords a stretched isomer (10E-anti), which thermally reforms the coiled starting meterial at room temperature via a sequence of tautomerization and C? C rotation. Heating or ultrasonic treatment can reverse this sequence and drive it farther to populate another stretched isomer (10E-syn) which is thermally stable at room temperature. This stretched form aggregates (presumably to dimers) already at concentrations at which the coiled species still appears to be fully monomeric.     

Application of Sputter-Assisted EPMA to Depth Profile Analysis

 A common problem in depth profile measurement is the calibration of the depth scale. The new technique of sputter assisted electron probe microanalysis offers the possibility of calculating the composition as well as the depth scale solely from the acquired X-ray intensity data without further information, e.g. sputter rates. To achieve a depth resolution that is smaller than the depth of information of the electron probe, i.e. 0.1–1 μm, special deconvolution algorithms must be applied to the acquired data. To assess the capabilities of this new technique it was applied to a Ti/Al/Ti multilayer on Si under different measurement conditions. Quantitative depth profiles were obtained by application of a deconvolution algorithm based on maximum entropy analysis. By comparison of these profiles with AES depth profiles and AFM roughness measurements, it was shown that the limiting factor to the achievable depth resolution is the occurrence of surface roughening induced by the sputtering process rather than the relatively large depth of information of the electron probe. We conclude that for certain applications sputter-assisted EPMA can be regarded as a valid depth profiling technique with a depth resolution in the nm range.     

Spectrofluorimetric Determination of Methyl Paraben in Pharmaceutical Preparations by Means of its Dansyl Chloride Derivative

 A spectrofluorimetric method for the determination of methyl paraben based on derivatization with the labelling reagent dansyl chloride (DNS-Cl), is presented. The effect of the reaction variables (pH, DNS-Cl concentration, temperature, reaction time) and instrumental parameters, has been examined. A linear calibration graph in the ng/ml range has been established. The limit of detection is 18 ng/ml with relative standard deviation less than 3%. The proposed method has been satisfactorily applied to determination of the paraben in two pharmaceutical preparations. Received May 25, 1999. Revision October 20, 1999.     

Über die katalytische Wirkung einiger Elemente in einer Landolt-Reaktion auf “Brom-Grundlage”; Ultramikrobestimmung von Molybdän(VI)

Zusammenfassung Die Landolt-Reaktion auf Bromgrundlage des Systems Wasserstoffperoxid—Bromid—Ascorbinsäure—Tolidin wird durch Mo(VI), Fe(III), Cu(II), V(V) und W(VI) katalysiert. Auf dieser Grundlage läßt sich Mo(VI) mit Hilfe der Simultankomparationsmethode bei 65° C in einem Konzentrationsbereich von 1 bis 10g/5 ml und bei 50° C in einem Konzentrationsbereich von 0,1 bis 1g/5 ml mit einem relativen Fehler von ±0.5% bestimmen.

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Summary The Landolt reaction on a bromine basis of the system hydrogen peroxide-bromide-ascorbic acid-tolidine is catalyzed by Mo(VI), Fe(III), Cu(II), V(V), and W(VI). On this basis, Mo(VI) can be determined with the aid of the simultaneous comparation method at 65° C in a concentration range from 1 to 10g/5 ml and at 50° C in a concentration range from 0.1 to 1g/5 ml with a relative error of ±0.5%.

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Résumé La «réaction de Landolt, avec brome» du système eau oxygénée—bromure—acide ascorbique—tolidine, est catalysée par Mo-VI, Fe-III, Cu-II, V-V et W-VT. On peut donc doser Mo-VI par la méthode de comparaison simultanée à 65° C dans un domaine de concentration de 1 à 10g/5 ml et à 50° C de 0,1 à 1g/5 ml avec une erreur relative de ±0,5%.

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Die vorliegende Arbeit sollte Herrn Prof. Dr.E. Schulek zu seinem 70. Geburtstag gewidmet werden. Noch ehe die Publikation möglich war, ist Herr Prof.Schulek am 14. Oktober 1964 leider gestorben.     

Validation of <Superscript>239,240</Superscript>Pu, <Superscript>238</Superscript>Pu separation method using molecular recognition product AnaLig<Superscript>®</Superscript> Pu02 gel and extraction chromatography TRU<Superscript>®</Superscript> resin

Two separation techniques for plutonium determination using AnaLig^® Pu02 molecular recognition technology product (MRT) and extraction chromatography TRU^® resin were tested. The methods performance was investigated by analysis of National Physical Laboratory (NPL-Alpha-Beta High, ABH 2003, 2005) intercomparison test samples. The results obtained for both procedures were compared in terms of activities and recoveries. Data analysis showed good agreement with the reference values. The AnaLig^® Pu02 separation method for ^239,240Pu, ²³⁸Pu determination was successfully validated with the same performance as the TRU^® resin method.