Detection and discrimination of spectral peaks and notches at 1 and 8 kHz

The ability of subjects to detect and discriminate spectral peaks and notches in noise stimuli was determined for center frequencies fc of 1 and 8 kHz. The signals were delivered using an insert earphone designed to produce a flat frequency response at the eardrum for frequencies up to 14 kHz. In experiment I, subjects were required to distinguish a broadband reference noise with a flat spectrum from a noise with either a peak or a notch at fc. The threshold peak height or notch depth was determined as a function of bandwidth of the peak or notch (0.125, 0.25, or 0.5 times fc). Thresholds increased with decreasing bandwidth, particularly for the notches. In experiment II, subjects were required to detect an increase in the height of a spectral peak or a decrease in the depth of a notch as a function of bandwidth. Performance was worse for notches than for peaks, particularly at narrow bandwidths. For both experiments I and II, randomizing (roving) the overall level of the stimuli had little effect at 1 kHz, but tended to impair performance at 8 kHz, particularly for notches. Experiments III-VI measured thresholds for detecting changes in center frequency of sinusoids, bands of noise, and spectral peaks or notches in a broadband background. Thresholds were lowest for the sinusoids and highest for the peaks and notches. The width of the bands, peaks, or notches had only a small effect on thresholds. For the notches at 8 kHz, thresholds for detecting glides in center frequency were lower than thresholds for detecting a difference in center frequency between two steady sounds. Randomizing the overall level of the stimuli made frequency discrimination of the sinusoids worse, but had little or no effect for the noise stimuli. In all six experiments, performance was generally worse at 8 kHz than at 1 kHz. The results are discussed in terms of their implications for the detectability of spectral cues introduced by the pinnae.     

Evaluation of a Gelatin-Modified Poly(ε-Caprolactone) Film as a Scaffold for Lung Disease

Lung transplantation is a necessary step for the patients with the end stage of chronic obstructive pulmonary disease. The use of artificial lungs is a promising alternative to natural lung transplantation which is complicated and is restricted by low organ donations. For successful lung engineering, it is important to choose the correct combination of specific biological cells and a synthetic carrier polymer. The focus of this study was to investigate the interactions of human lung epithelial cell line NCl-H292 that is involved in lung tissue development with the biodegradable poly(ε-caprolactone) before and after its chemical modification to evaluate potential for use in artificial lung formation. Also, the effect of polymer chemical modification on its mechanical and surface properties has been investigated. The poly(ε-caprolactone) surface was modified using aminolysis followed by immobilization of gelatin. The unmodified and modified polymer surfaces were characterized for roughness, tensile strength, and NCl-H292 metabolic cell activity. The results showed for the first time the possibility for NCI-H292 cells to adhere on this polymeric material. The resazurin assay showed that the metabolic activity at 24?h postseeding of 80% in the presence of the unmodified and greater than 100% in the presence of the modified polymer was observed. The roughness of the poly(ε-caprolactone) increased from 4 to 26?nm and the film strength increased from 0.01 to 0.045 kN when the material was chemically modified. The results obtained to date show potential for using modified poly(ε-caprolactone) as a scaffold for lung tissue engineering.     

Phase-stepped ESPI and moiré interferometry for measuring stress-intensity factor andJ integral

Electronic-speckle-pattern interferometry and moiré interferometry have been used to calculateK ₁ andJ for compact tension specimens. Automated-fringe-pattern analysis enables the full-field of data to be used with the minimum of operator intervention. Measurements are shown to be accurate to within 10 percent. TheJ-measurement procedure employed could form the basis of an automatic-fault detection system.     

Efficient preparation of 2,4-methanoproline

Using a modification of the route described by Clardy and Hughes et al., 2,4-methanoproline hydrochloride (1) was prepared in four steps and 70% overall yield from dl-serine methyl ester.     

Catalytic mechanism of Golgi-resident human tyrosylprotein sulfotransferase-2: A mass spectrometry approach

Human tyrosylprotein sulfotransferases catalyze the transfer of a sulfuryl moiety from the universal sulfate donor PAPS to the hydroxyl substituent of tyrosine residues in proteins and peptides to yield tyrosine sulfated products and PAP. Tyrosine sulfation occurs in the trans-Golgi network, affecting an estimated 1% of the tyrosine residues in all secreted and membrane-bound proteins in higher order eukaryotes. In this study, an effective LC-MS-based TPST kinetics assay was developed and utilized to measure the kinetic properties of human TPST-2 and investigate its catalytic mechanism when G protein-coupled CC-chemokine receptor 8 (CCR8) peptides were used as acceptor substrates. Through initial rate kinetics, product inhibition studies, and radioactive-labeling experiments, our data strongly suggest a two-site ping-pong model for TPST-2 action. In this mechanistic model, the enzyme allows independent binding of substrates to two distinct sites, and involves the formation of a sulfated enzyme covalent intermediate. Some insights on the important amino acid residues at the catalytic site of TPST-2 and its covalent intermediate are also presented. To our knowledge, this is the first detailed study of the reaction kinetics and mechanism reported for human TPST-2 or any other Golgi-resident sulfotransferase.     

Chemical-genetic screen identifies riluzole as an enhancer of Wnt/β-catenin signaling in melanoma

To identify new protein and pharmacological regulators of Wnt/β-catenin signaling, we used a cell-based reporter assay to screen a collection of 1857 human-experienced compounds for their ability to enhance activation of the β-catenin reporter by a low concentration of WNT3A. This identified 44 unique compounds, including the FDA-approved drug riluzole, which is presently in clinical trials for treating melanoma. We found that treating melanoma cells with riluzole in?vitro enhances the ability of WNT3A to regulate gene expression, to promote pigmentation, and to decrease cell proliferation. Furthermore riluzole, like WNT3A, decreases metastases in a mouse melanoma model. Interestingly, siRNAs targeting the metabotropic glutamate receptor, GRM1, a reported indirect target of riluzole, enhance β-catenin signaling. The unexpected regulation of β-catenin signaling by both riluzole and GRM1 has implications for the future uses of this drug.     

Quinol-containing ligands enable high superoxide dismutase activity by modulating coordination number,charge, oxidation states and stability of manganese complexes throughout redox cycling

Reactivity assays previously suggested that two quinol-containing MRI contrast agent sensors for H₂O₂, [Mn(H2qp1)(MeCN)]²⁺ and [Mn(H4qp2)Br₂], could also catalytically degrade superoxide. Subsequently, [Zn(H2qp1)(OTf)]⁺ was found to use the redox activity of the H2qp1 ligand to catalyze the conversion of O₂˙⁻ to O₂ and H₂O₂, raising the possibility that the organic ligand, rather than the metal, could serve as the redox partner for O₂˙⁻ in the manganese chemistry. Here, we use stopped-flow kinetics and cryospray-ionization mass spectrometry (CSI-MS) analysis of the direct reactions between the manganese-containing contrast agents and O₂˙⁻ to confirm the activity and elucidate the catalytic mechanism. The obtained data are consistent with the operation of multiple parallel catalytic cycles, with both the quinol groups and manganese cycling through different oxidation states during the reactions with superoxide. The choice of ligand impacts the overall charges of the intermediates and allows us to visualize complementary sets of intermediates within the catalytic cycles using CSI-MS. With the diquinolic H4qp2, we detect Mn(iii)-superoxo intermediates with both reduced and oxidized forms of the ligand, a Mn(iii)-hydroperoxo compound, and what is formally a Mn(iv)-oxo species with the monoquinolate/mono-para-quinone form of H4qp2. With the monoquinolic H2qp1, we observe a Mn(ii)-superoxo ↔ Mn(iii)-peroxo intermediate with the oxidized para-quinone form of the ligand. The observation of these species suggests inner-sphere mechanisms for O₂˙⁻ oxidation and reduction that include both the ligand and manganese as redox partners. The higher positive charges of the complexes with the reduced and oxidized forms of H2qp1 compared to those with related forms of H4qp2 result in higher catalytic activity (k_cat ∼ 10⁸ M⁻¹ s⁻¹ at pH 7.4) that rivals those of the most active superoxide dismutase (SOD) mimics. The manganese complex with H2qp1 is markedly more stable in water than other highly active non-porphyrin-based and even some Mn(ii) porphyrin-based SOD mimics.

Manganese complexes with polydentate quinol-containing ligands are found to catalyze the degradation of superoxide through inner-sphere mechanisms. The redox activity of the ligand stabilizes higher-valent manganese species.     

Cucurbit[7]uril Disrupts Aggregate Formation Between Rhodamine B Dyes Covalently Attached to Glass Substrates

Dye aggregation is detrimental to the performance of high optical density dye-doped photonic materials. To overcome this challenge, the ability of cucurbit[7]uril (CB7) as a molecular host to disrupt aggregate formation on glass substrates was examined. Rhodamine B was covalently attached to glass slides by initially coating the surface with azidohexylsiloxane followed by copper-catalyzed “click” triazole formation with rhodamine B propargyl ester. The absorption and emission spectra of rhodamine B coated slides in water indicated diverse heterogeneous properties as surface dye density varied. Fluorescence quenching due to dye aggregation was evident at high surface dye density. Addition of aqueous cucurbit[7]uril (CB7) to the surface-tethered dyes perturbed the spectra to reveal a considerable reduction in heterogeneity, which suggested that the presence of a surface in close proximity does not significantly impair CB7’s ability to complex with tethered rhodamine B.     

The effect of hearing loss on the resolution of partials and fundamental frequency discrimination

The relationship between the ability to hear out partials in complex tones, discrimination of the fundamental frequency (F0) of complex tones, and frequency selectivity was examined for subjects with mild-to-moderate cochlear hearing loss. The ability to hear out partials was measured using a two-interval task. Each interval included a sinusoid followed by a complex tone; one complex contained a partial with the same frequency as the sinusoid, whereas in the other complex that partial was missing. Subjects had to indicate the interval in which the partial was present in the complex. The components in the complex were uniformly spaced on the ERB(N)-number scale. Performance was generally good for the two "edge" partials, but poorer for the inner partials. Performance for the latter improved with increasing spacing. F0 discrimination was measured for a bandpass-filtered complex tone containing low harmonics. The equivalent rectangular bandwidth (ERB) of the auditory filter was estimated using the notched-noise method for center frequencies of 0.5, 1, and 2 kHz. Significant correlations were found between the ability to hear out inner partials, F0 discrimination, and the ERB. The results support the idea that F0 discrimination of tones with low harmonics depends on the ability to resolve the harmonics.