REDUCED PHOTOINACTIVATION OF 10-DODECYL ACRIDINE ORANGE-SENSITIZED YEAST CELLS AT HIGH FLUENCE RATES MEASUREMENTS AND COMPUTER SIMULATIONS

Abstract-During the development of a photodamage cell sorter several photosensitizers were tested for their ability to photoinactivate more than 90% of the sensitized cells after a brief irradiation with a fluence of 10 kJ/m². In pilot experiments, yeast cells sensitized with 10-dodecyl acridine orange (DAO) were effectively photoinactivated after receiving a fluence of 10 kJ/m² delivered in 8 s. However, when the same fluence was delivered in 3 μ s during passage through a focused laser beam in the cell sorter, all cells survived.
Computer simulations of the relevant photophysical and chemical reactions inside the irradiated cell were used to investigate the cause of this phenomenon. The results indicated that the absence of photoinactivation by DAO, after flash irradiations, was caused by the combined effects of (1) limited oxygen diffusion into the cell and (2) a reduced number of collisions between photosensitizer triplet and oxygen molecules during the irradiation due to saturation of the intracellular photosensitizer triplet concentration. The contributions of triplet-triplet annihilation and triplet quenching by ground state photosensitizer molecules were found to be minimal and not significant. These findings indicate that Type II photosensitizers are incapable of rapid selective photoinactivation in cell sorters.     

Extraction of proteins from plant tissues for two-dimensional electrophoresis analysis

To increase the number of proteins detectable by two-dimensional electrophoresis (2-DE) in plants, we present a new procedure for extracting total proteins from plant tissue. This method avoids any loss of proteins in the course of sample preparation and results in two different fractions, one comprising mainly the cytoplasmatic proteins, the other one containing predominantly structure bond proteins. 2-DE patterns obtained from these two fractions show that the total number of different protein spots detected exceeds the degree of resolution commonly reported for plant proteins threefold.     

Analytical pyrolysis of carbohydrates: I. Chemical interpretation of matrix influences on pyrolysis-mass spectra of amylose using pyrolysis-gas chromatography-mass spectrometry

Inorganic additives, viz. Na₂CO₃, NaOH, HCl, ZnC₂, NaH₂PO₄, Na₂HPO₄, NaCl. MgSO₄ and sea salt, to the pyrolysis matrix change the pyrolysis-mass spectrum of amylose significantly. Carbonyl compounds, acids and their lactones, furans, pyranones anhydrosugars and aromatic substances are found in different ratios under the various conditions, as determined by pyrolysis-gas chromatography-mass spectrometry (Py-GC-MS).Carbonyl compounds, acids and lactones are released from alkaline and neutral matrices. Furans and anhydrohexoses are particularly formed under neutral and acidic conditions. Pyranones are specific for phosphate matrices. Unsaturated hydrocarbons and aromatic substances arise from strongly alkaline or dehydrating matrices. Degradation pathways are proposed for various compound categories.The results of pyrolysis-mass spectrometry and Py-GC-MS are highly compatible.     

Highly efficient recycling of a "sandwich" type polyoxometalate oxidation catalyst using solvent resistant nanofiltration

A "sandwich" type polyoxometalate catalyst ([MeN(n-C8H17)3]12[WZn3(ZnW9O34)2]) was very efficiently recycled by nanofiltration with almost quantitative retention, using an alpha-alumina supported mesoporous gamma-alumina membrane.     

The solid-phase Nicholas reaction: scope and limitations

Two libraries of alpha-substituted alkynes has been prepared on solid phase using a sequential Sonogashira/Nicholas reaction approach. The scope of nucleophiles in the Nicholas reaction on solid phase has been investigated, including carbon, oxygen, nitrogen, sulfur, fluoride, and hydride nucleophiles. The conditions for the reaction sequence have been optimized in terms of Lewis acid, catalyst for the Sonogashira step, temperature, reaction time, and decomplexation method, enabling the five-step sequence to be performed in 1 day.     

Rearrangements of 3-heterobicyclo[3.2.0]hept-6-enes

Studies directed at a synthesis of dihydrothiepin 1b have resulted in the elucidation of several factors which effect cyclobutene ring opening in the 3-heterobicyclo[3.2.0]hept-6-ene ring system. We report the unexpected rearrangement of 4a, 4b, 13b and 13c to the synthetically useful a-vinyl-2,5-dihydrothiophenes 7a, 7b, 15a and 15b, respectively. Conversion of 4a to 6 is suggested to occur by a 1,3-rearrangement of 4a to isomeric 3-thiabicyclo[3.2.0]hept-6-ene 19 followed by cyclobutene ring opening in 19.     

Molecular Structure of 1,3,5-Tris (Trimethylstannyl) Benzene: An Electron Diffraction Study

The molecular structure of 1,3,5-tris (trimethylstannyl) benzene has been determined by gas-phase electron diffraction. The C — C bond length is in good agreement with that in benzene. In agreement with the somewhat electron-releasing character of the substituents, the endocyclic bond angles at the substituents are somewhat smaller than 120°. The mean value of Sn — C bond lengths is greater than that in tetraphenyltin and tetramethyltin. The SnMe₃ groups appear freely rotating around the C_aryl — Sn bonds. The following bond lengths (r _g) and bond angles were determined: (Sn — C)_mean 2.150 ± 0.007 Å, C — C 1.399 ± 0.005 Å, (C — H)_mean 1.105 ± 0.006 Å, < C — C(Sn) — C 117.7 ± 1.7º, < C_aryl — Sn — C_methyl 106.7 ± 0.7º < Sn — C — H 111.5 ± 0.9º.     

Mechanism of hierarchical porosity development in MFI zeolites by desilication: the role of aluminium as a pore-directing agent

The role of the concentration and the nature of aluminium in the creation of hierarchical porosity in both commercial and synthesized MFI zeolites have been investigated through controlled mesoporosity development by desilication in alkaline medium. Framework aluminium controls the process of framework silicon extraction and makes desilication selective towards intracrystalline mesopore formation. An optimal molar Si/Al ratio in the range 25-50 has been identified; this leads to an optimal mesoporosity centred around 10 nm and mesopore surface areas of up to 235 m(2) g(-1) while preserving the intrinsic crystalline and acidic properties. At lower framework Si/Al ratios the relatively high Al content inhibits Si extraction and hardly any mesopores are created, while in highly siliceous ZSM-5 unselective extraction of framework Si induces formation of large pores. The existence of framework Al sites in different T positions that are more or less susceptible to the alkaline treatment, and the occurrence of re-alumination, are tentative explanations for the remarkable behaviour of Al in the desilication process. The presence of substantial extra framework Al, obtained by steam treatment, inhibits Si extraction and related mesopore formation; this is attributed to re-alumination of the extraframework Al species during the alkaline treatment. Removal of extraframework Al species by mild oxalic acid treatment restores susceptibility to desilication, which is accompanied by formation of larger mesopores due to the enhanced Si/Al ratio in the acid-treated zeolite.     

The effect of (natural) polyols on the initial colour of heavy metal- and zinc-free poly(vinyl chloride)

The propensity of (natural) polyols dispersed in heavy metal- and zinc-free PVC sheets to improve the initial colour, e.g. the colour during the first minutes of exposure to high processing temperatures (short-term stability), has been investigated. It is shown using W(Lab) values that the initial colour improves upon addition of polyols containing primary hydroxyl groups. The polyols act as HCl scavengers, presumably via an acid-catalysed S_N2 substitution of the primary hydroxyl groups by chloride ions. In contrast, polyols with only secondary or tertiary hydroxyl groups accelerate the thermal degradation of PVC. Notwithstanding, the efficacy of the (natural) polyols containing primary hydroxyl groups will be reduced if the polyol is susceptible to competitive acid-catalysed intramolecular cyclodehydration reactions under the processing conditions. This is substantiated by a comparison of the behaviour of mannitol and 3,4-di-O-methyl-d-mannitol. The methylated derivative, which is less prone to undergo intramolecular cyclodehydration, improves the initial colour of heavy metal- and zinc-free PVC sheets more significantly than mannitol itself.     

Spectroscopy and quantum chemical modeling reveal a predominant contribution of excitonic interactions to the bathochromic shift in alpha-crustacyanin, the blue carotenoprotein in the carapace of the lobster Homarus gammarus

To resolve the molecular basis of the coloration mechanism of alpha-crustacyanin, we used (13)C-labeled astaxanthins as chromophores for solid-state (13)C NMR and resonance Raman spectroscopy of [6,6',7,7']-(13)C(4) alpha-crustacyanin and [8,8',9,9',10,10',11,11',20,20']-(13)C(10) alpha-crustacyanin. We complement the experimental data with time-dependent density functional theory calculations on several models based on the structural information available for beta-crustacyanin. The data rule out major changes and strong polarization effects in the ground-state electron density of astaxanthin upon binding to the protein. Conformational changes in the chromophore and hydrogen-bond interactions between the astaxanthin and the protein can account only for about one-third of the total bathochromic shift in alpha-crustacyanin. The exciton coupling due to the proximity of two astaxanthin chromophores is found to be large, suggesting that aggregation effects in the protein represent the primary source of the color change.