Reduction in accumulation of [3H]triphenylmethylphosphonium cation in neuroblastoma cells caused by optical probes of membrane potential: Evidence for interactions between carbocyanine dyes and lipophilic anions

The accumulation of [³H]triphenylmethylphosphonium cation in neuroblastoma N1E 115 cells in the presence of tetraphenylboron is reduced by 3,3′-diethylthiadicarbocyanine iodide and by 3,3′-dipropylthiadicarbocyanine iodide. This reduction in uptake of the lipophilic cation is not due to the carbocyanine dyes depolarizing the plasma membrane of these cells but due to an interaction between the carbocyanine dyes and tetraphenylboron leaving less of the lipophilic anion free in solution to assist uptake of the lipophilic cation. This interaction is shown to have a 1:1 stoicheiometry.     

固定对组织光学性质的影响

利用带有积分球的SHIMADZU UV—240分光光度计,测量了组织在固定前和后的反射率和透射率,分析了福尔马林固定对组织光学性质的影响。     

Signalling fitness: larger males sire more offspring. Studies of the stalk-eyed fly Cyrtodiopsis whitei (Diopsidae,Diptera)

Some species of stalk-eyed flies (Diopsidae, Diptera) have a sexual dimorphism of eyespan. For example, Cyrtodiopsis whitei males have much longer eyestalks than females of equal body length. Expression of this trait increases with the bearers' size. The slope of the log-log regression line eyespan versus body length is close to two in males, while in females it is roughly one. Behavioural experiments suggest that male eyespan signals quantitatively a male's strength or attractiveness to a competitor or mate. We used 3 pure strains of C. Whitei, which were distinguished by their different phosphoglucomutase allele outfit. We compared the reproductive success of males of different sizes and found the number of offspring to be directly proportional to body length. Thus eyespan, rising with the square of body length, provides an exaggerated and highly conclusive signal in the advertisement of fitness.     

Activity, isoenzymes and purity of mushroom tyrosinase in commercial preparations

Several lots of commercial tyrosinase preparations were examined with regard to their enzyme activity, isoenzyme composition and purity. Enzyme activity toward catechol, -dopa and tyrosine showed significant variations from lot to lot and activation by SDS. Distribution of isoenzyme forms also varied from lot to lot. Comparisons of electrophoretic and isoelectric focusing protein profiles showed considerable differences and distributions of the proteins in each sample. Tyrosinase appeared to be a minor component in each preparation when compared to a partially purified enzyme. Investigators using commercial tyrosinase should exercise caution in interpreting data due to the presence of different isoenzyme forms, their distribution in various lots, and the presence of numerous other proteins.     

Fumonisin B1 — Isolation and analytical characterisation for the application as reference material

A procedure for the production of Fumonsin B1 with a purity of more than 97% has been worked out. The isolated material was characterised by NMR and GC-MS and showed good agreement to literature data, thus proving the identity of the analysed FB1. For the purity assessment with LC-FLD and LC-MS a commercially available standard and the isolated FB1 sample were analysed under the same conditions. The overall purity of the FB₁ samples was estimated as 91.9±8.1% (reference) and 97.6±2.4% (biopure), respectively. Presented at the 25^th Mykotoxin Workshop in Giessen, Germany, May 19–21, 2003     

Identification of the optical isomers of the amino acids in Polysphondylium violaceum acrasin

Using optically specific enzymes on the hydrolysate of glorin, we show the structure of natural glorin to be: N-propionyl-γ-L-glutamyl-L-ornithine-δ-lactam ethyl ester     

Perturbations of membrane structure by optical probes: II. Differential scanning calorimetry of dipalmitoyllecithin and its analogs interacting with merocyanine 540

Summary Differential scanning calorimetry of multilamellar liposomes, interacting with the optical probe Merocyanine 540, yields quantitative information about perturbances of the bilayer structure induced by this dye. At low dye: lipid ratios, the dye perturbs primarily its own microenvironment, which is laterally separated from the unmodified lipid domain and exhibits modified thermotropic properties. A further increase in the dye concentration results in a perturbance of the whole lipid bilayer. The degree of perturbance is sensitive to structural modifications in the head-group region of the lipids. It is concluded that Merocyanine 540 reports in every case, even at infinite dilution, on localized events originating from a perturbed microenvironment.An abstract of this paper was presented at the XI Congress of Biochemistry, Toronto, in July 1979.     

A global benchmark study using affinity-based biosensors

To explore the variability in biosensor studies, 150 participants from 20 countries were given the same protein samples and asked to determine kinetic rate constants for the interaction. We chose a protein system that was amenable to analysis using different biosensor platforms as well as by users of different expertise levels. The two proteins (a 50-kDa Fab and a 60-kDa glutathione S-transferase [GST] antigen) form a relatively high-affinity complex, so participants needed to optimize several experimental parameters, including ligand immobilization and regeneration conditions as well as analyte concentrations and injection/dissociation times. Although most participants collected binding responses that could be fit to yield kinetic parameters, the quality of a few data sets could have been improved by optimizing the assay design. Once these outliers were removed, the average reported affinity across the remaining panel of participants was 620 pM with a standard deviation of 980 pM. These results demonstrate that when this biosensor assay was designed and executed appropriately, the reported rate constants were consistent, and independent of which protein was immobilized and which biosensor was used.     

Analytical Approach to the Prism Coupling Problem in the Kretschmann Configuration

Prism coupling in the Kretschmann configuration is a well-known method for excitation of surface plasmon polaritons (SPP’s) in a metal film bounded from one side by a prism and from the other side by air. The analysis of the reflectance in the upper medium (prism) is based on the well-known Fresnel’s formula. Due to the fact that this formula cannot be inverted directly to give the complex dielectric permittivity and the thickness of the metal film from measured values of the reflectance at three or more different angles of incidence, an additional analysis is needed. Here, such an analysis is presented. The special case of illumination with He–Ne laser (λ = 632.8 nm) of silver film bounded by air is considered. A new asymptotic formula for the Lorentz dip is derived. Our experimental data for silver film are reported too.