A comparison of the fluorescence dynamics of single molecules of a green fluorescent protein: one- versus two-photon excitation.

We report on the dynamics of fluorescence from individual molecules of a mutant of the wild-type green fluorescent protein (GFP) from Aequorea victoria, super folder GFP (SFGFP). SFGFP is a novel and robust variant designed for in vivo high-throughput screening of protein expression levels. It shows increased thermal stability and is able to retain its fluorescence when fused to poorly folding proteins. We use a recently developed single-molecule technique which combines fluorescence-fluctuation spectroscopy and time-correlated single photon counting in order to characterize the photophysical properties of SFGFP under one- (OPE) and two- (TPE) photon excitation conditions. We use Rhodamine 110 as a model chromophore to validate the methodology and to explain the single-molecule results of SFGFP. Under OPE, single SFGFP molecules undergo fluorescence flickering on the time scale of micros and tens of micros due to triplet formation and ground-state protonation-deprotonation, respectively, as demonstrated by excitation intensity- and pH-dependent experiments. OPE single-molecule fluorescence lifetimes indicate heterogeneity in the population of SFGFP, indicating the presence of the deprotonated I and B forms of the SFGFP chromophore. TPE of single SFGFP molecules results in the photoconversion of the chromophore. TPE of single SFGFP molecules show fluorescence flickering on the time scale of micros due to triplet formation. A flicker connected with protonation-deprotonation of the SFGFP chromophore is detected only at low pH. Our results show that SFGFP is a promising fusion reporter for intracellular applications using OPE and TPE microscopy.     

Investigations of Steric and Electronic Factors Influencing the Cyclopalladation of meta-Toluidine Analogues

The cyclopalladation of two different types of aniline derivatives is described: the acetylated anilines N-(3-methylphenyl)acetamide ( 2a ), 3-(acetylamino)-4-chlorobenzoic acid ( 2c ), and N-(2-chlorophenyl)acetamide ( 2d ) are cyclometalated easily with palladium(II) acetate and trifluoroacetic acid to yield the corresponding complexes 4a, 4c , and 4d , respectively, whereas the acetylated meta-toluidine N-(2-chloro-5-methylphenyl)acetamide ( 2b ) cannot be metalated at the only accessible site between the acetylamino and the methyl group. This aromatic C? H bond can be activated, however, with the second type of meta-toluidine derivatives: the 2-chloro-5, N-dimethyl-N-nitrosoaniline ( 3b ) readily undergoes cyclopalladation to yield the corresponding Pd^II complex di-μ-trifluoro-acetato-bis[3-chloro-6-methyl-2-(N-methyl-N- nitrosoamino)phenyl-C,N? O]dipalladium(II) ( 5b ) containing a five-membered palladacycle with coordination of Pd^II at the nitroso N-atom, which is established by ¹⁵N-NMR spectroscopy.     

A general bound on the critical temperature of Ising ferromagnets

An inequality relating the spin correlation functions for a general Ising ferromagnet is found. This inequality gives an improved upper bound for the critical temperature of translationally invariant ferromagnets with pair-wise interactions.     

Effect of CO2 Flow Rate on the Extraction of Astaxanthin and Fatty Acids from Haematococcus pluvialis Using Supercritical Fluid Technology

Haematococcus pluvialis is the largest producer of natural astaxanthin in the world. Astaxanthin is a bioactive compound used in food, feed, nutraceutics, and cosmetics. In this study, astaxanthin extraction from H. pluvialis by supercritical fluid extraction was evaluated. The effects of temperature (40 and 50 °C), pressure (40 and 50 MPa), and CO₂ flow rate (2 and 4 L/min) were investigated. The results showed that the highest astaxanthin recovery was obtained at 50 °C/50 MPa and the CO₂ flow rates evaluated had no significant effect. It was possible to achieve astaxanthin recoveries of 95% after 175 min for a CO₂ flow rate of 2 L/min, and 95 min for CO₂ flow rate of 4 L/min. The ω-6/ω-3 ratios obtained were similar in all conditions, reaching 0.87, demonstrating that the extracts from H. pluvialis by SFE are rich in unsaturated fatty acids (UFA) which increases their positive effects when used as a functional ingredient in food.     

Antimony speciation in soils: improving the detection limits using post-column pre-reduction hydride generation atomic fluorescence spectroscopy (HPLC/pre-reduction/HG-AFS)

HG-AFS is highly sensitive and low cost detection system and its use for antimony chemical speciation coupled to HPLC is gaining popularity. However speciation analysis in soils is strongly hampered because the most efficient extractant reported in the literature (oxalic acid) strongly inhibits the generation of SbH₃ by Sb(V), the major species in this kind of matrix, severely affecting its detection limits. The purpose of this research is to reduce the detection limit of Sb(V), by using a post column on-line reduction system with l-cysteine reagent (HPLC/pre-reduction/HG-AFS). The system was optimized by experimental design, optimum conditions found were 2% (w/v) and 10 °C temperature coil. Detection limits of Sb(V) and Sb(III) in oxalic acid (0.25 mol L⁻¹) were improved from 0.3 and 0.1 μg L⁻¹ to 0.07 and 0.07 μg L⁻¹, respectively. The methodology developed was applied to Chilean soils, where Sb(V) was the predominant species.     

History of Vinyl Chloride Polymers

In 1926 Semon tried to dehydrohalogenate high molecular weight poly(vinyl chloride) (PVC) in a high boiling solvent to get an unsaturated polymer which might bond rubber to metal. Unexpectedly, he obtained plasticized PVC, a flexible product inert both electrically and chemically. This discovery opened the door to the commercialization of PVC, a plastic with an annual United States production now exceeding 6 billion pounds. Special PVC's and PVC products have been developed taking advantage of the many favorable properties. Rigid structural products from house siding to pipes are becoming of increasing importance. Two main types of polymers have been utilized: 1) one prepared by suspension polymerization, and 2) a special variety prepared by colloidal polymerization and spray drying. This latter material has been especially useful for making plastisols. Plasticizers and stabilizers were developed to maximize useful and nontoxic properties. Vinyl chloride monomer (VCM) production and co-polymerization evolved as lower cost processes, higher quality products, and greater manufacturing safety were introduced. Recent challenges for the industry have included pollution and carcinogenic hazards which have been overcome by imaginative new technologies. The rate of growth of the industry is shown graphically.