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Physical estimation of triplet phases--effects of different radiation sources and modes for profile scans.

A comparative study has been made of the intensity profiles from three-beam experiments to estimate triplet phases using radiation from a conventional sealed-tube X-ray source and two different synchrotron sources. Synchrotron radiation, with its much smaller angular divergence, narrower spectral bandwidth and higher flux, distinctly improves the experimental conditions for physical phase estimation. Pure psi scans about the primary diffraction vector, such as can be made with a six-circle diffractometer, further improve the conditions compared with combined omega/psi scans with a four-circle instrument, where the rotation in psi is accomplished by combining rotations about the three axes omega, chi and varphi. Interference profiles collected by pure psi scans and unfocused synchrotron radiation have FWHM values reduced by factors in the range 20-35 relative to those obtained with combined omega-2theta/psi scans and radiation from a conventional source. As a consequence of these changes, which also involve greatly increased peak amplitudes, the 0/pi-type asymmetry of the profiles is exposed much more pronounced closer to the three-beam point, enabling unambiguous phase assignment for all triplets that were studied. The superiority of the pure psi scan will be even more important in studies of general phases for which the phase information lies also in the relative heights of the (sharp) interference maxima for a triplet and the Friedel-related triplet.

Journal Article↗

Triplet-state kinetics of Zn-porphyrin cytochrome c in micellar media. Measurement of intermicellar exchange rates.

The interactions of protein molecules with surfactant assemblies in aqueous and hydrocarbon media have been studied via the triplet-state kinetics of Zn-porphyrin cytochrome c in solutions containing an anionic [sodium bis(2-ethylhexyl)sulfosuccinate, AOT] or a cationic (cetyltrimethylammonium bromide, CTAB) surfactant. In aqueous solution, the observed triplet state decay is single exponential with a lifetime of 8 ms. In aqueous solutions of AOT and in AOT-reversed micellar solutions, biexponential triplet state decays were observed, indicating that interactions between the surfactant and the protein occur, resulting in a change in protein conformation near the porphyrin ring. In CTAB-reversed micellar solutions, quenching of the Zn-porphyrin cytochrome c triplet state by ferricyanide and methyl viologen was studied. Because the quenching is exchange-limited under the conditions used, the exchange rate constants for the water pools can be obtained from these experiments. The observed exchange rate constants are in the range (1-5) x 10(7) M-1 S-1, depending on the water content of the reversed micelles and on the type of quencher used. These values are three orders of magnitude lower than the calculated collision rate of the reversed micelles.

Cytochrome c Group↗

Anthralin-derived transients--I. The triplet state and the products of its reaction with oxygen in benzene.

Direct laser excitation in benzene of 1,8-dihydroxy-9-anthrone (anthralin) does not lead to transient species with lifetimes in the nanosecond time regime or longer. The triplet state has been produced in benzene by pulse radiolysis and characterised in terms of its absorption spectrum (lambda max 560 nm), natural lifetime (11 microseconds), self-quenching properties (kmicrosecond = 2.6 x 10(7) l mol-1 s-1) and triplet energy (234 kJ mol-1). There is no tendency in the non-polar medium for production of either the triplet or ground state in a tautomeric form. The observed triplet state reacts with oxygen with a typical rate constant, 2.2 x 10(9) l mol-1 s-1. The products of this reaction are singlet oxygen (approximately 64%) and the anthralin radical (approximately 14%).

Anthralin↗

Nanosecond kinetic absorption and calorimetric studies of cyclopentenone: the triplet, self-quenching, and the predimerization biradicals.

The photolysis of 2-cyclopentenone has been studied by a combination of kinetic absorption spectrophotometry and time-resolved photoacoustic calorimetry. The lifetime of the cyclopentenone triplet is strongly concentration dependent and corresponds to a value of 380 +/- 75 ns at infinite dilution in acetonitrile. The biradical intermediate (or pair of isomeric biradicals) immediately preceding formation of photodimer forms with very high efficiency upon quenching of triplet cyclopentenone by a second ground state cyclopentenone molecule and has an energy of 47 kcal/mol relative to two molecules of reactant. Quenching of the cyclopentenone triplet by conjugated dienes is much slower than expected, which is the reason that early estimates of the triplet lifetime were much too short.

Calorimetry↗

An unstable triplet repeat in a gene related to myotonic muscular dystrophy.

Synthetic oligonucleotides containing GC-rich triplet sequences were used in a scanning strategy to identify unstable genetic sequences at the myotonic dystrophy (DM) locus. A highly polymorphic GCT repeat was identified and found to be unstable, with an increased number of repeats occurring in DM patients. In the case of severe congenital DM, the paternal triplet allele was inherited unaltered while the maternal, DM-associated allele was unstable. These studies suggest that the mutational mechanism leading to DM is triplet amplification, similar to that occurring in the fragile X syndrome. The triplet repeat sequence is within a gene (to be referred to as myotonin-protein kinase), which has a sequence similar to protein kinases.

Amino Acid Sequence↗

Oculopharyngeal muscular dystrophy: a point mutation which mimics the effect of the PABPN1 gene triplet repeat expansion mutation.

BACKGROUND: Oculopharyngeal muscular dystrophy (OPMD) is an autosomal dominant late onset neuromuscular disease characterised by proximal muscle weakness, ptosis, and swallowing difficulty. The only causative mutation described to date is a triplet repeat expansion consisting of two to seven additional base triplets in a repeat sequence in exon 1 of the polyadenine binding protein nuclear 1 (PABPN1) gene. This results in an increase in length of a polyalanine tract in the PABPN1 protein from 10 to 12-17 residues. OBJECTIVE: Description of another mutation in a case of OPMD. METHODS: Sequence analysis of exon 1 of the PABPN1 gene was undertaken on 202 patients referred for a possible diagnosis of OPMD but negative for the triplet repeat expansion mutation. RESULTS: A case was identified with typical symptoms of OPMD, negative for the repeat expansion mutation but with a missense mutation in PABPN1 close to the 3' end of the normal polyalanine codon repeat sequence. CONCLUSIONS: The single base mutation changes a glycine codon to an alanine codon and results in an increase in the number of contiguous polyalanine codons. This mimics the effect of the common triplet repeat expansion mutation and represents a previously undescribed mechanism of mutation.

Aged↗

Evolution of a triplet repeat in a conifer.

The opportunity to trace the evolution of a triplet repeat is rare, especially for seed-plant lineages with a well-defined fossil record. Microsatellite PtTX2133 sequences from 18 species in 2 conifer genera were used to calibrate the birth of a CAGn repeat, from its protomicrosatellite origins to its repeat expansion. Birth occurred in the hard-pine genome ~ 136 million years ago, or 14 million generations ago, then expanded as a polymorphic triplet repeat 136-100 million years before a major North American vicariance event. Calibration of the triplet-repeat birth and expansion is supported by the shared allelic lineages among Old and New World hard pines and the shared alleles solely among North American diploxylon or hard pines. Five CAGn repeat units appeared to be the expansion threshold for Old and New World diploxylon pines. Haploxylon pine species worldwide did not undergo birth and repeat expansion, remaining monomorphic, with a single imperfect 198-bp allele. A sister genus, Picea, had only a region of cryptic simplicity, preceding a proto-microsatellite region. The polymorphic triplet repeat in hard pines is older than some long-lived microsatellites reported for reptiles, yet younger than those reported for insects. Some cautionary points are raised about phylogenetic applications for this long-lived microsatellite.

Alleles↗

Stillbirth rates in singletons, twins and triplets in Sweden, 1869 to 2001.

The temporal variation in the stillbirth rates (SBR), measured as the number of stillborn per 1000 total births, among singletons, twins and triplets was studied on Swedish birth data for the period 1869 to 2001 and comparisons with data from other populations were made. Among both single and multiple births there were marked, almost monotonously decreasing trends in the stillbirth rates. Among singletons the stillbirth rate decreased from 29.5 per 1000 in the period 1869 to 1878 to 3.4 in the period 1991 to 2001. Among twins the stillbirth rate decreased from 94 per 1000 in 1869 to 1878 to a minimum of 8.2 in 1991 to 2001 and among triplets from 166 per 1000 to a minimum of 19.8. The relative declining pattern in the SBRs was almost the same, being 88% among singletons, 91% among twins and 88% among triplets. In the 1980s and 1990s the definition of the stillbirth rate was changed in many countries, including Finland, but no changes in the definition of stillbirths have been made in Sweden. The effect of the artificial reproduction techniques, including in vitro fertilization, on the rates of multiple maternities is also discussed. It was noted especially that they had a more marked effect on the triplet than on the twinning rate.

Female↗

Quenching of cascade reaction between triplet and photochrome probes with nitroxide radicals. A novel labeling method in study of membranes and surface systems.

We proposed a new method for the study of molecular dynamics and fluidity of the living and model biomembranes and surface systems. The method is based on the measurements of the sensitized photoisomerization kinetics of a photochrome probe. The cascade triplet cis-trans photoisomerization of the excited stilbene derivative sensitized with the excited triplet Erythrosin B has been studied in a model liposome membrane. The photoisomerization reaction is depressed with nitroxide radicals quenching the excited triplet state of the sensitizer. The enhanced fluorescence polarization of the stilbene probe incorporated into liposome membranes indicates that the stilbene molecules are squeezed in a relatively viscous media of the phospholipids. Calibration of the "triple" cascade system is based on a previously proposed method that allows the measurement of the product of the quenching rate constant and the sensitizer's triplet lifetime, as well as the quantitative detection of the nitroxide radicals in the vicinity of the membrane surface. The experiment was conducted using the constant-illumination fluorescence technique. Sensitivity of the method using a standard commercial spectrofluorimeter is about 10(-12) mol of fluorescence molecules per sample and can be improved using an advanced fluorescence technique. The minimal local concentration of nitroxide radicals or any other quenchers being detected is about 10(-5) M. This method enables the investigation of any chemical and biological surface processes of microscopic scale when the minimal volume is about 10(-3) microL or less.

Biosensing Techniques↗

Triplets of spikes in a model of spike timing-dependent plasticity.

Classical experiments on spike timing-dependent plasticity (STDP) use a protocol based on pairs of presynaptic and postsynaptic spikes repeated at a given frequency to induce synaptic potentiation or depression. Therefore, standard STDP models have expressed the weight change as a function of pairs of presynaptic and postsynaptic spike. Unfortunately, those paired-based STDP models cannot account for the dependence on the repetition frequency of the pairs of spike. Moreover, those STDP models cannot reproduce recent triplet and quadruplet experiments. Here, we examine a triplet rule (i.e., a rule which considers sets of three spikes, i.e., two pre and one post or one pre and two post) and compare it to classical pair-based STDP learning rules. With such a triplet rule, it is possible to fit experimental data from visual cortical slices as well as from hippocampal cultures. Moreover, when assuming stochastic spike trains, the triplet learning rule can be mapped to a Bienenstock-Cooper-Munro learning rule.

Action Potentials↗

Photophysical properties of the lowest excited singlet and triplet states of thio- and seleno-psoralens

The decay processes of the lowest excited singlet and triplet states of five heteropsoralens (HPS) were investigated by steady-state and shift-phase fluorometry and by laser-flash photolysis in different solvents. The emission spectra of HPS are detectable only in trifluoroethanol (TFE), where fluorescence lifetimes (tau F) and quantum yields (phi F) were measured. The triplet lifetimes (tau T), triplet (phi T) and singlet-oxygen production (phi delta) quantum yields were determined in benzene, ethanol and TFE by laser-flash photolysis. Semiempirical (INDO/1-CI) calculations allowed the nature of the lowest excited singlet and triplet states and transition probabilities to be obtained. Theoretical and experimental results indicate that the two lowest excited singlet states S1 and S2 of HPS are close-lying and different in nature (pi,pi* and n,pi*). The "proximity effect" between these two states controls the photophysical properties of HPS as it does for the other furocoumarins. However, HPS have a peculiar behavior with respect to the related compounds because they are fluorescent and have, in three cases, detectable intersystem crossing only in TFE. This behavior can be tentatively explained by a different energy gap and/or order between the S1 and S2 states.

Journal Article↗

Does the photochemical conversion of colchicine into lumicolchicines involve triplet transients? A solvent dependence study.

beta- and gamma-lumicolchicines are photoproducts formed by the cycloisomerization of the tropolone ring of colchicine (COL) alkaloids. The mechanism of the photoconversion, suggested to involve the triplet state, is examined here by studying the effect of the solvent polarity on the lumicolchicine photoisomer ratio. Triplet COL, detected by laser flash photolysis, is quenched by oxygen, but not by transtilbene or 1-methylnaphtalene. Neither the quantum yield of conversion of COL nor the photoproduct ratio was altered by the presence of oxygen. Likewise, energy transfer to COL from triplet acetone produced by either isobutanal/horseradish peroxidase system or tetramethyldioxetane thermolysis failed to provoke photoreaction of COL. Our data argue against the intermediacy of a COL triplet state in the photoisomerization and stress on the role of specific solvent-solute interactions in determining the partitioning of excited singlet state into the beta- and gamma-isomer formation.

Colchicine↗

A laser flash photolysis study of fenofibric acid in aqueous buffered media: unexpected triplet state inversion in a derivative of 4-alkoxybenzophenone.

Laser excitation of aqueous solutions of fenofibric acid (FA) at pH 7.4 show the formation of two reaction intermediates, the triplet state and the hydrated electron. The former is longer lived in water than in acetonitrile; its anionic form decays irreversibly by decarboxylation to give a carbanion that protonates before or after rearrangement. Several spectroscopic and quenching studies suggest that in aqueous media the triplet state of FA has a pi,pi* character, in comparison with an n,pi* character in organic media. Further, the known chemistry of the triplet, including decarboxylation and hydrogen abstraction, occurs predominantly from the n,pi* state, and as a consequence, activation energies are higher when the lowest triplet has a pi,pi* character. Photoionization is more important in aqueous than in organic media and involves a biphotonic process. Hydrated electrons are trapped by FA, leading to the corresponding ketyl radical after protonation.

Benzophenones↗

Study of the interaction between triplet riboflavin and the alpha-, betaH- and betaL-crystallins of the eye lens.

Time-resolved photolysis studies of riboflavin (RF) were carried out in the presence and absence of alpha-, betaH- and betaL-crystallins of bovine eye lens. The transient absorption spectra, recorded 5 micros after the laser pulse, reveal the presence of the absorption band (625-675 nm) of the RF neutral triplet state (tau = 42 micros) accompanied by the appearance of a long-lived absorption (tau = 320 micros) in the 500-600 nm region due to the formation of the semireduced RF radical. The RF excited state is quenched by the crystallin proteins through a mechanism that involves electron transfer from the proteins to the flavin, as shown by the decrease of the triplet RF band with the concomitant increase of the band of its semireduced form. Tryptophan loss on RF-sensitized photooxidation of the crystallins when irradiated with monochromatic visible light (450 nm) in a 5% oxygen atmosphere was studied. A direct correlation was found between the triplet RF quenching rate constants by the different crystallin fractions and the decomposition rate constants for the exposed and partially buried tryptophans in the proteins. The RF-sensitized photooxidation of the crystallins is accompanied by the decrease of the low molecular weight constituents giving rise to its multimeric forms. A direct correlation was observed between the initial rate of decrease of the low molecular weight bands corresponding to the irradiated alpha-, betaH- and betaL-crystallins and the quenching constant values of triplet RF by the different crystallins. The correlations found in this study confirm the importance of the Type-I photosensitizing mechanism of the crystallins, when RF acts as a sensitizer at low oxygen concentration, as can occur in the eye lens.

Animals↗

Alpha-methylstilbene and the duality of mechanism in the quenching of stilbene triplets by molecular oxygen.

Spin-exchange quenching of alpha-methylstilbene triplets by molecular oxygen and by the free radical di-tert-butyl nitroxide is shown to favor the cis isomer more than does natural decay. The effect of the two quenching events is an identical 7% decrease in the fraction of perpendicular triplets that decay to the trans isomer. The conclusion that relaxed stilbene triplets reside in a shallow minimum corresponding to a geometry in which the two benzyl moieties are orthogonal was based on the observation that their quenching by O2 does not alter the trans/cis photostationary ratio. Our results with alpha-methylstilbene confirm the hypothesis that in the case of stilbene spin exchange quenching by O2 at the twisted geometry favors the cis isomer but occurs in competition with excitation transfer from transoid triplets that leads to the trans isomer and to singlet oxygen. The opposite effects of the two oxygen quenching paths on stilbene isomer composition cancel accidentally leading to an overall insensitivity of benzophenone-sensitized photostationary states to the presence of oxygen. Quenching rate constants derived on the basis of this cancellation are close to diffusion-controlled and predict singlet O2 quantum yields of 0.08 and 0.13 in the presence of air and under an O2 atmosphere, in good agreement with experimental measurements.

Molecular Conformation↗

A comparison of word-recognition abilities assessed with digit pairs and digit triplets in multitalker babble.

This study compares, for listeners with normal hearing and listeners with hearing loss, the recognition performances obtained with digit-pair and digit-triplet stimulus sets presented in multitalker babble. Digits 1 through 10 (excluding 7) were mixed in approximately 1,000 ms segments of babble from 4 to -20 dB signal-to-babble (S/B) ratios, concatenated to form the pairs and triplets, and recorded on compact disc. Nine and eight digits were presented at each level for the digit-triplet and digit-pair paradigms, respectively. For the listeners with normal hearing and the listeners with hearing loss, the recognition performances were 3 dB and 1.2 dB better, respectively, on digit pairs than on digit triplets. For equal intelligibility, the listeners with hearing loss required an approximately 10 dB more favorable S/B than the listeners with normal hearing. The distributions of the 50% points for the two groups had no overlap.

Adult↗

Accounting for triplet and saturation effects in FCS measurements.

Fluorescence correlation spectroscopy (FCS) is an increasingly important tool for determining low concentrations and dynamics of molecules in solution. Oftentimes triplet transitions give rise to fast blinking effects, which are accounted for by including an exponential term in the fitting of the autocorrelation function (ACF). In such cases, concomitant saturation effects also modify the amplitude and shape of the remaining parts of the ACF. We review studies of triplet and saturation effects in FCS and present a simple procedure to obtain more accurate results of particle concentrations and diffusional dynamics in experiments where triplet kinetics are evident, or where moderate laser powers approaching saturation levels are used, for example, to acquire sufficient photon numbers when observation times are limited. The procedure involves use of a modified function for curve-fitting the ACF, but there are no additional fitting parameters. Instead, a simple calibration of the total fluorescence count rate as a function of relative laser power is fit to a polynomial, and the non-linear components of this fit, together with the relative laser power used for the FCS measurement, are used to specify the magnitude of additional terms in the fitting function. Monte Carlo simulations and experiments using Alexa dyes and quantum dots, with continuous and pulsed laser excitation, demonstrate the application of the modified fitting procedure with first order correction terms, in the regime where distortions in the ACF due to photobleaching and detector dead time are small compared to those of fluorescence saturation and triplet photophysics.

Algorithms↗

Triplet chemotherapy with gemcitabine, a platinum, and a third agent in the treatment of advanced non-small cell lung cancer.

Cisplatin-based chemotherapy regimens were shown to prolong survival, relieve symptoms, and improve quality of life in a cost-effective manner in patients with advanced non-small cell lung cancer. New chemotherapeutic agents introduced in the 1990s, such as gemcitabine, paclitaxel, docetaxel, vinorelbine, and irinotecan, were shown to be as effective or more effective than cisplatin. The doublet combination of these new agents with cisplatin or carboplatin was more active than either as single agents, and randomized trials showed that new combinations such as gemcitabine/ cisplatin, vinorelbine/cisplatin, and paclitaxel/cisplatin were preferred to cisplatin alone or to etoposide/cisplatin. Gemcitabine has a mild toxicity profile that allows it to be combined easily in triplet combinations with other doublets. Phase I-II trials of such triplet combinations show that each drug can be given safely in full dosage. Preliminary efficacy results of these gemcitabine-based triplet combinations are encouraging. Randomized trials comparing triplets such as gemcitabine/paclitaxel/carboplatin with the corresponding doublets are indicated.

Antineoplastic Combined Chemotherapy Protocols↗