Frequency-frequency correlation functions and apodization in two-dimensional infrared vibrational echo spectroscopy: a new approach

Ultrafast two-dimensional infrared (2D-IR) vibrational echo spectroscopy can probe structural dynamics under thermal equilibrium conditions on time scales ranging from femtoseconds to approximately 100 ps and longer. One of the important uses of 2D-IR spectroscopy is to monitor the dynamical evolution of a molecular system by reporting the time dependent frequency fluctuations of an ensemble of vibrational probes. The vibrational frequency-frequency correlation function (FFCF) is the connection between the experimental observables and the microscopic molecular dynamics and is thus the central object of interest in studying dynamics with 2D-IR vibrational echo spectroscopy. A new observable is presented that greatly simplifies the extraction of the FFCF from experimental data. The observable is the inverse of the center line slope (CLS) of the 2D spectrum. The CLS is the inverse of the slope of the line that connects the maxima of the peaks of a series of cuts through the 2D spectrum that are parallel to the frequency axis associated with the first electric field-matter interaction. The CLS varies from a maximum of 1 to 0 as spectral diffusion proceeds. It is shown analytically to second order in time that the CLS is the T(w) (time between pulses 2 and 3) dependent part of the FFCF. The procedure to extract the FFCF from the CLS is described, and it is shown that the T(w) independent homogeneous contribution to the FFCF can also be recovered to yield the full FFCF. The method is demonstrated by extracting FFCFs from families of calculated 2D-IR spectra and the linear absorption spectra produced from known FFCFs. Sources and magnitudes of errors in the procedure are quantified, and it is shown that in most circumstances, they are negligible. It is also demonstrated that the CLS is essentially unaffected by Fourier filtering methods (apodization), which can significantly increase the efficiency of data acquisition and spectral resolution, when the apodization is applied along the axis used for obtaining the CLS and is symmetrical about tau=0. The CLS is also unchanged by finite pulse durations that broaden 2D spectra.     

Deprotonation dynamics and stokes shift of pyranine (HPTS)

The short and intermediate time scale dynamics of the photoacid pyranine (1-hydroxy-3,6,8-pyrenetrisulfonic acid, commonly referred to as HPTS) are studied with visible pump-probe spectroscopy in various solvents to elucidate the nature of its proton-transfer kinetics in water. The observed time dependences of HPTS are compared with those of the methoxy derivative, MPTS. A global fitting procedure is employed to model both the spectral shift (Stokes shift) caused by solvent reorganization and deprotonation of pyranine in water. Three distinct time-dependent features can be clearly identified. They are the Stokes shift (1 ps in H(2)O and 1.5 ps in D(2)O), followed by the deprotonation processes, which gives rise to a biexponential decay of the protonated species with time constants (in H(2)O) of 3 and 88 ps. By the use of a model previously discussed in the literature, the biexponential process can be interpreted as an initial deprotonation step followed by the longer time scale process which separates the resulting ion pair. The results presented here are consistent with some of the previous reports but unambiguously identify and quantitatively measure the Stokes shift as a separate and distinct phenomenon from the deprotonation process, in contrast to other reports that have suggested that all short time (a few picoseconds) dynamics are merely a Stokes shift.     

Probing dynamics of complex molecular systems with ultrafast 2D IR vibrational echo spectroscopy

Ultrafast 2D IR vibrational echo spectroscopy is described and a number of experimental examples are given. Details of the experimental method including the pulse sequence, heterodyne detection, and determination of the absorptive component of the 2D spectrum are outlined. As an initial example, the 2D spectrum of the stretching mode of CO bound to the protein myoglobin (MbCO) is presented. The time dependence of the 2D spectrum of MbCO, which is caused by protein structural evolution, is presented and its relationship to the frequency-frequency correlation function is described and used to make protein structural assignments based on comparisons to molecular dynamics simulations. The 2D vibrational echo experiments on the protein horseradish peroxidase are presented. The time dependence of the 2D spectra of the enzyme in the free form and with a substrate bound at the active site are compared and used to examine the influence of substrate binding on the protein's structural dynamics. The application of 2D vibrational echo spectroscopy to the study of chemical exchange under thermal equilibrium conditions is described. 2D vibrational echo chemical exchange spectroscopy is applied to the study of formation and dissociation of organic solute-solvent complexes and to the isomerization around a carbon-carbon single bond of an ethane derivative.     

Analysis of Voice and Quantitative Measurement of Glottal Gap After Thyroplasty Type I in the Treatment of Unilateral Vocal Paralysis

Thyroplasty type I is one of several surgical treatments in which improving the voice of unilateral vocal fold paralysis is the ultimate objective. The goal of the surgery is the medialization of the paralyzed vocal fold. The purpose of this study is to evaluate the effectiveness of thyroplasty type I through acoustical analysis, aerodynamic measures, and quantitative videostroboscopic measurements. We report on 20 patients with unilateral vocal cord paralysis who underwent thyroplasty type I. We performed preoperative and postoperative video image analysis (normalized glottal gap area) and computer-assisted voice analysis (fundamental frequency, jitter, shimmer, noise-to-harmonic ratio, mean phonation time, mean flow rate, mean subglottic pressure) in all patients. The glottal gap was significantly reduced after thyroplasty type I. Postoperative voice quality was characterized by an improved pitch and amplitude pertubation (jitter and shimmer), phonation time (mean phonation time), and subglottic pressure (mean subglottic pressure). Thyroplasty type I is an effective method for regaining glottal closure and vocal function.     

Laryngeal Gestures During Stop Production Using High-Speed Digital Images

On acoustic and fiberscopic studies of stop consonants, voice onset time and glottal width have been shown to be greatest in heavily aspirated stops, next greater for slightly aspirated stops, and least for unaspirated stops. Integrated activity of the thyroarytenoid and posterior cricoarytenoid muscles has been reported to be involved in differentiating aspirate characteristics of the stops. However, the fine movement of mucosal edges of vocal folds during the production of stops has not been well documented. In recent years, a new method for high-speed digital recording of laryngeal dynamics has made this possible. In the current study, the movements of vocal fold edges were documented during the period of stop production using a fiberscopic system of high-speed digital images. By observing the glottal width and the visual vibratory movements of vocal folds before voice onset, the heavily aspirated stop was characterized as being more prominent and dynamic than the slightly aspirated and unaspirated stops.     

Hyaluronic Acid: Its Role in Voice

The extracellular matrix (ECM), once regarded simply as a structural scaffold, is now recognized as an important modulator of cellular behavior and function. One component that plays a prominent role in this process is hyaluronic acid (HA)--a molecule found in many different tissues. Research into the roles of HA indicates that it plays a key role in tissue viscosity, shock absorption, and space filling. Specifically, research into the role of HA in laryngology indicates that it has profound effects on the structure and viscosity of vocal folds. This article provides an introduction to the structure and biological functions of HA and its importance in voice. In addition, an overview of the pharmaceutical applications of HA is discussed.     

Vocal Nodules and Laryngeal Morphology

Our purpose was to study the occurrence of vocal fold nodules under conditions of habitual vocal abuse associated with increased laryngeal muscle tension, to identify the existence of a relationship between vocal nodules and laryngeal morphology. We studied one group of 30 subjects with vocal nodules, 18 to 50 years old, who were compared with two control groups, one of females and one of males, consisting of 30 subjects each. The parameters evaluated were: type of vocal folds coaptation, glottic proportion (GP) and abduction angle (AA), obtained by videotelelaryngoscopy. In the nodules group, the larynges presented a mean value of GP similar to that of the female group, both of which were lower than the mean GP value of the male group. On the other hand, the mean AA was lower than the one in the female group, and closer to the one in the male group. We concluded that vocal nodules were present only in larynges with a predominantly young female morphology, with functional limitations of abduction.     

Conformational dynamics and stability of HP35 studied with 2D IR vibrational echoes

Two-dimensional infrared (2D IR) vibrational echo spectroscopy was used to measure the fast dynamics of two variants of chicken villin headpiece 35 (HP35). The CN of cyanophenylalanine residues inserted in the hydrophobic core were used as a vibrational probe. Experiments were performed on both singly (HP35-P) and doubly CN-labeled peptide (HP35-P(2)) within the wild-type sequence, as well as on HP-35 containing a singly labeled cyanophenylalanine and two norleucine mutations (HP35-P NleNle). There is a remarkable similarity between the dynamics measured in singly and doubly CN-labeled HP35, demonstrating that the presence of an additional CN vibrational probe does not significantly alter the dynamics of the small peptide. The substitution of two lysine residues by norleucines markedly improves the stability of HP35 by replacing charged with nonpolar residues, stabilizing the hydrophobic core. The results of the 2D IR experiments reveal that the dynamics of HP35-P are significantly faster than those of HP35-P NleNle. These observations suggest that the slower structural fluctuations in the hydrophobic core, indicating a more tightly structured core, may be an important contributing factor to HP35-P NleNle's increased stability.     

Quantification of acitretin in human plasma by microbore liquid chromatography-negative chemical ionization mass spectrometry

A highly sensitive liquid chromatographic-mass spectrometric procedure has been developed to quantitate plasma concentrations of acitretin, a dermatologic agent used to treat severe psoriasis. The assay utilizes the combination of normal-phase microbore high-performance liquid chromatography, negative chemical ionization mass spectrometry, selective ion monitoring and stable isotope dilution. The method has been used to measure acitretin and its metabolite, 13-cis-acitretin, over a range of 1-20 ng/ml in human plasma. The inter-assay precision was 5.3% for acitretin and 3.9% for 13-cis-acitretin, while the intra-assay precisions for acitretin and 13-cis-acitretin were 10.8 and 12.7%, respectively. Reproducibility of the assay for acitretin and 13-cis-acitretin, which was determined by the relative standard deviation of multiple analyses of the same quality assurance sample, was 5.9 and 8.1%, respectively.     

Photoinduced electron transfer and geminate recombination for photoexcited acceptors in a pure donor solvent

Photoinduced electron transfer and geminate recombination are studied for the systems rhodamine 3B (R3B(+)) and rhodamine 6G (R6G(+)), which are cations, in neat neutral N,N-dimethylaniline (DMA). Following photoexcitation of R3B(+) or R6G(+) (abbreviated as R(+)), an electron is transferred from DMA to give the neutral radical R and the cation DMA(+). Because the DMA hole acceptor is the neat solvent, the forward transfer rate is very large, approximately 5x10(12) s(-1). The forward transfer is followed by geminate recombination, which displays a long-lived component suggesting several percent of the radicals escape geminate recombination. Spectrally resolved pump-probe experiments are used in which the probe is a "white" light continuum, and the full time-dependent spectrum is recorded with a spectrometer/charge-coupled device. Observations of stimulated emission (excited state decay-forward electron transfer), the R neutral radical spectrum, and the DMA(+) radical cation spectrum as well as the ground-state bleach recovery (geminate recombination) make it possible to unambiguously follow the electron transfer kinetics. Theoretical modeling shows that the long-lived component can be explained without invoking hole hopping or spin-forbidden transitions.