Isolation and sequence determination of peptides in the venom of the spider wasp (Cyphononyx dorsalis) guided by matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry

Micro-scale (sub-pmol) isolation and sequence determination of three peptides from the venom of the solitary spider wasp Cyphononyx dorsalis is described. We isolated two novel peptides Cd-125 and Cd-146 and a known peptide Thr(6)-bradykinin from only two venom sacs of solitary spider wasp Cyphononyx dorsalis without bioassay-guided fractionation. but instead guided by MALDI-TOF MS. The MALDI-TOF MS analysis of each fraction showed the purity and molecular weight of the components, which led to the isolation of the peptides virtually without loss of sample amount. The sequences of the novel peptides Cd-125 (Asp-Thr-Ala-Arg-Leu-Lys-Trp-His) and Cd-146 (Ser-Glu-Thr-Gly-Asn-Thr-Val-Thr-Val-Lys-Gly-Phe-Ser-Pro-Leu-Arg) were determined by Edman degradation together with mass spectrometry. and finally corroborated by solid-phase synthesis. The known peptide Thr(6)-bradykinin (Arg-Pro-Pro-Gly-Phe-Thr-Pro-Phe-Arg) was identified by comparison with the synthetic authentic specimen. This is the first example for any kinins to be found in Pompilidae wasp venoms. The procedure reported here can be applicable to studies on many other components of solitary wasp venoms with limited sample availability. (C) 2001 Elsevier B.V. Ltd. All rights reserved.

DIODE-LASER AND FOURIER-TRANSFORM SPECTROSCOPY OF THE (CH3OH)-C-13 C-O STRETCHING MODE

In this work we present high resolution Doppler limited absorption spectra measurements of the C-O stretching mode of (CH3OH)-C-13, obtained from diode laser spectroscopy, and the Fourier Transform spectrum obtained at 0. 12 cm-1 resolution. By using these data and previously known spectroscopic information, we determined the frequency and the J quantum number for the multiplets of the P and R(J) branches of the C-O stretching fundamental band. Infrared transitions in coincidence with emission lines of the regular CO2 laser and some of its isotope parents are pointed out.

Characterization of gallotannins from Astronium species by flow injection analysis-electrospray ionization-ion trap-tandem mass spectrometry and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

An in vitro thermal analysis during different light-activated hydrogen peroxide bleaching

This study measured the critical temperature reaching time and also the variation of temperature in the surface of the cervical region and within the pulp chamber of human teeth submitted to dental bleaching using 35% hydrogen peroxide gel activated by three different light sources. The samples were randomly divided into 3 groups (n = 15), according to the catalyst light source: Halogen Light (HL), High Intensity Diode Laser (DL), and Light Emmited Diode (LED). The results of temperature variation were submitted to the analysis of variance and Tukey test with p < 0.05. The temperature increase (mean value and standard deviation) inside the pulp chamber for the HL group was 6.8 ± 2.8°C; for the DL group was 15.3 ± 8.8°C; and for the LED group was 1.9 ± 1.0°C for. The temperature variation (mean value and standard deviation) on the tooth surface, for the group irradiated with HL was 9.1 ± 2.2°C; for the group irradiated with DL were 25.7 ± 18.9°C; and for the group irradiated with LED were 2.6 ± 1.4°C. The mean temperature increase values were significantly higher for the group irradiated with DL when compared with groups irradiated with HL and LED (p < 0.05). When applying the inferior limits of the interval of confidence of 95%, an application time of 38.7 s was found for HL group, and 4.4 s for DL group. The LED group did not achieve the critical temperatures for pulp or the periodontal, even when irradiated for 360 s. The HL and DL light sources may be used for dental bleaching for a short period of time. The LED source did not heat the target tissues significantly within the parameters used in this study. © 2010 Pleiades Publishing, Ltd.

Two-wavelength whole-field interferometry setup for thermal lens study

In this paper we present a new approach for thermal lens analysis using a two-wavelength DSPI (Digital Speckle Pattern Interferometry) setup for wavefront sensing. The employed geometry enables the sensor to detect wavefronts with small phase differences and inherent aberrations found in induced lenses. The wavefronts was reconstructed by four-stepping fringe evaluation and branch-cut unwrapping from fringes formed onto a diffusive glass. Real-time single-exposure contour interferograms could be obtained in order to get discernible and low-spacial frequency contour fringes and obtain low-noise measurements. In our experiments we studied the thermal lens effect in a 4% Er-doped CaO-Al2O3 glass sample. The diode lasers were tuned to have a contour interval of around 120 μm. The incident pump power was longitudinally and collinearly oriented with the probe beams. Each interferogram described a spherical-like wavefront. Using the ABCD matrix formalism we obtained the induced lens dioptric power from the thermal effect for different values of absorbed pump power. © 2012 Copyright SPIE.

Effects of rotation in the laser-pulse photoassociation in a thermal gas of atoms

Photoassociation is a possible route for the formation of chemical bonds. In this process, the binding of colliding atoms can be induced by means of a laser field. Photoassociation has been studied in the ultracold regime and also with temperatures well above millikelvins in the thermal energy domain, which is a situation commonly encountered in the laboratory. A photoassociation mechanism can be envisioned based on the use of infrared pulses to drive a transition from free colliding atoms on the electronic ground state to form a molecule directly on that state. This work takes a step in this direction, investigating the laser-pulse-driven formation of heteronuclear diatomic molecules in a thermal gas of atoms including rotational effects. Based on the assumption of full system controllability, the maximum possible photoassociation yield is deduced. The photoassociation probability is calculated as a function of the laser parameters for different temperatures. Additionally, the photoassociation yield induced by subpicosecond pulses of a priori fixed shape is compared to the maximum possible yield.

Identification of Corynebacterium spp. isolated from bovine intramammary infections by matrix-assisted laser desorption ionization-time of flight mass spectrometry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Laser irradiation and thermal treatment inducing selective crystallization in Sb2O3-Sb2S3 glassy films

The influence of both thermal treatment and laser irradiation on the structural and optical properties of films in the Sb 2 O 3 –Sb 2 S 3 system was investigated. The films were prepared by RF-sputtering using glass compositions as raw materials. Irreversible photodarkening effect was observed after exposure the films to a 458nm solid state laser. It is shown, for the first time, the use of holographic technique to measure “in situ”, simultaneously and independently, the phase and amplitude modulations in glassy films. The films were also photo-crystallized and analysed “in situ” using a laser coupled to a micro-Raman equipment. Results showed that Sb 2 S 3 crystalline phase was obtained after irradiation. The effect of thermal annealing on the structure of the films was carried out. Different from the result obtained by irradiation, thermal annealing induces the crystallization of the Sb 2 O 3 phase. Photo and thermal induced effects on films were studied using UV–Vis and Raman spectroscopy, atomic force microscopy (AFM), thermal analysis (DSC), X-ray diffraction, scanning electron microscopy (MEV) and energy-dispersive X-ray spectroscopy (EDX).

Degree of conversion of different composite resins photo-activated with light-emitting diode and argon ion laser

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effectiveness of the diode laser in the treatment of ligature-induced periodontitis in rats: a histopathological, histometric, and immunohistochemical study

The aim of this study was to evaluate the effects of the use of a high-power gallium-aluminum-arsenide diode laser (GaAlAs; 808 nm, 1 W, 20 s, 20 Hz, 10 J) alone or as adjunctive therapy to scaling and root planing in the treatment of induced periodontitis in rats. Periodontitis was induced by placing a ligature around the mandibular first molar of 60 rats. After 7 days, the ligature was removed and the animals were divided into four groups as follows: C (control), no periodontal treatment; SRP, scaling and root planing (SRP); DL, diode laser (DL) irradiation treatment; and SRP/DL, both SRP and DL irradiation treatment. Five animals from each group were euthanized at 7, 15, and 30 days posttreatment. The effectiveness of the treatments was evaluated in the furcation area using histopathological analysis, histometric analysis of alveolar bone loss (ABL), and immunohistochemical detection of tartrate-resistant acid phosphatase (TRAP), runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN). DL, alone or in combination with adjunctive therapy to SRP in the treatment of experimental periodontitis, resulted in a decreased local inflammatory response. At 7-days posttreatment, the DL and SRP/DL groups had fewer TRAP-positive cells and more RUNX2-positive cells. There was greater OCN immunolabeling in the DL group than in the C and SRP groups at 15 days. There was less ABL in the DL and SRP/DL groups at 15 and 30 days. In conclusion, DL was effective in the treatment of ligature-induced periodontitis in rats, both when used alone and when used as adjunctive therapy to SRP.

Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Differentiation of the Dimorphic Fungal Species Paracoccidioides brasiliensis and Paracoccidioides lutzii

Isolates of Paracoccidioides brasiliensis and Paracoccidioides lutzii, previously characterized by molecular techniques, were identified for the first time by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). All isolates were correctly identified, with log score values of >2.0. Thus, MALDI-TOF MS is a new tool for differentiating species of the genus Paracoccidioides.

Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Characterization of Primary Amine End-Functionalized Polystyrene and Poly(methyl methacrylate) Synthesized by Living Anionic Polymerization Techniques

The reaction of living anionic polymers with 2,2,5,5-tetramethyl-1-(3-bromopropyl)-1-aza-2,5- disilacyclopentane (1) was investigated using coupled thin layer chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Structures of byproducts as well as the major product were determined. The anionic initiator having a protected primary amine functional group, 2,2,5,5-tetramethyl- 1-(3-lithiopropyl)-1-aza-2,5-disilacyclopentane (2), was synthesized using all-glass high-vacuum techniques, which allows the long-term stability of this initiator to be maintained. The use of 2 in the preparation of well-defined aliphatic primary amine R-end-functionalized polystyrene and poly(methyl methacrylate) was investigated. Primary amino R-end-functionalized poly(methyl methacrylate) can be obtained near-quantitatively by reacting 2 with 1,1-diphenylethylene in tetrahydrofuran at room temperature prior to polymerizing methyl methacrylate at -78 °C. When 2 is used to initiate styrene at room temperature in benzene, an additive such as N,N,N',N'- tetramethylethylenediamine is necessary to activate the polymerization. However, although the resulting polymers have narrow molecular weight distributions and well-controlled molecular weights, our mass spectra data suggest that the yield of primary amine α-end-functionalized polystyrene from these syntheses is very low. The majority of the products are methyl α-end-functionalized polystyrene.