Chip gel electrophoresis as a tool for study of matrix metalloproteinase 9 interaction with metallothionein

Matrix metalloproteinases (MMPs) play numerous roles in physiological and pathological processes including cancer. Interactions of MMPs with other zinc-binding proteins are of great interest mainly from the point of view of the participation of these proteins in carcinogenesis. The aim of this study was to utilize chip gel electrophoresis for investigation of matrix metalloproteinase 9 (gelatinase B, MMP-9) interactions with collagen. The interaction was observed and the effect of temperature was tested. We further focused on the study of activation of MMP-9 by the presence of zinc-binding proteins called metallothioneins (MT). We confirmed the basic presumption of the activation of MMP-9 by the presence of MT similar to the temperature effect. Moreover, we chose the method of differential pulse voltammetry Brdicka reaction to verify our results. It can be concluded that the activity of MMP-9 was higher in the presence of MT in comparison with the experimental collagen(type I)-MMP-9 mixture in vitro.     

A model for the prediction of breathiness in vowels

The perception of breathiness in vowels is cued by multiple acoustic cues, including changes in aspiration noise (AH) and the open quotient (OQ) [Klatt and Klatt, J. Acoust. Soc. Am. 87(2), 820-857 (1990)]. A loudness model can be used to determine the extent to which AH masks the harmonic components in voice. The resulting "partial loudness" (PL) and loudness of AH ["noise loudness" (NL)] have been shown to be good predictors of perceived breathiness [Shrivastav and Sapienza, J. Acoust. Soc. Am. 114(1), 2217-2224 (2003)]. The levels of AH and OQ were systematically manipulated for ten synthetic vowels. Perceptual judgments of breathiness were obtained and regression functions to predict breathiness from the ratio of NL to PL (η) were derived. Results show that breathiness can be modeled as a power function of η. The power parameter of this function appears to be affected by the fundamental frequency of the vowel. A second experiment was conducted to determine if the resulting power function could estimate breathiness in a different set of voices. The breathiness of these stimuli, both natural and synthetic, was determined in a listening test. The model estimates of breathiness were highly correlated with perceptual data but the absolute predicted values showed some discrepancies.     

Assays for determination of matrix metalloproteinases and their activity

Matrix metalloproteinases (MMPs) are involved in many physiological and pathological processes. Due to their ability to cleave and to remodel components of surrounding tissues, MMPs may affect cell migration, differentiation, growth, inflammatory processes, neovascularization, wound healing, apoptosis, the uterine cycle and many other actions within the body, including those needed for tumorigenesis and other diseases.MMPs can therefore be used as potential markers for detecting various cancers, neurodegenerative, and immune and cardiovascular diseases. Numerous MMP assays were developed for clinical and research purposes, but far more attention has been devoted to understanding their biological functions.Due to differences in methodology, results obtained in various laboratory settings are difficult to compare because of the lack of standards and analytical methods of validation. Limits of detection of particular methods used for identifying MMPs are also disputable.Enzymatic, immunochemical and fluorimetric methods are particularly suitable for clinical use. In-vivo imaging methods offer many potential advantages in cancer research and diagnostics. Other methods are subject to investigation [e.g., phage display, multiple-enzyme/multiple-reagent assay system (MEMRAS) and activity-based profiling].     

Nonplanar Electron - Acoustic Shock Waves with Superthermal Hot Electrons

Nonplanar electron-acoustic shock waves having superthermal hot electrons are investigated with two temperature electrons model in unmagnetized plasma. Using reductive perturbation method, Korteweg-de Vries-Burgers (KdVB) equation is obtained in the cylindrical/spherical coordinates. Dissipation effect is introduced in the model by means of kinematic viscosity term. On the basis of the solutions of KdVB equation, variation of shock waves features (amplitude, velocity and width) with different plasma parameters are analysed. KdV-Burgers equation always leads to monotonic solitons and no oscillatory peak may appear. The combined effect of particle density (α), superthermal parameter (κ), electron temperature ratio (??) and kinetic viscosity (η₀) is numerically studied, and it is observed that these parameters significantly change the properties of the shock waves in nonplanar geometry especially in spherical coordinates. Results could be helpful to analyse the soliton features in laboratory as well as in the space environments.     

Direct measurement of the Chudakov effect

Experimental results for the restricted energy loss of pairs created from 1-178 GeV photons in a thin Au target and subsequently passing a CCD detector are presented. It is shown that pairs--when detected close to the creation vertex--suffer a reduced energy loss due to the internal screening of the charges constituting the pair. Furthermore, the ability to measure directly the energy of the pair by calorimetry enables a comparison with theory as a function of energy. The observed phenomenon is in good qualitative agreement with general expectations from the Chudakov effect but indicates a quantitative disagreement with either of two mutually disagreeing theories.     

Decay of three-particle high spin states in85Y

High spin states in⁸⁵Y have been excited in the reaction⁷²Ge(¹⁶O,p2n) at 48–60 MeV beam energy. From measurements ofnγ andγγ coincidences, excitation functions and angular distributions, the high spin spectrum of⁸⁵Y has been established up to 5.4 MeV excitation energy and spinI≦29/2. Lifetimes or limits of lifetimes have been determined for 14 levels via the recoil distance and Doppler shift attenuation method. Theg _9/2 decoupled proton band is found to backbend atI ^π=17/2⁺ as also seen in the reduction of the 17/2⁺→13/2⁺ and 21/2⁺→17/2⁺ E2 transition strengths. The lowest (3qp) positive parity band has predominantly aν ² g _9/2×πg _9/2 structure. The observed negative parity yrast states in the spin range 15/2^?≦I≦29/2^? are most probably of (g _9/2)² ×(f _5/2,p _1/2) (3qp) nature as suggested by a comparison of the⁸⁴Sr and⁸⁵Y level structures and electromagnetic transition strengths.     

Copper- and phosphine-free Sonogashira coupling reactions of aryl iodides catalyzed by an N,N-bis(naphthylideneimino)diethylenetriamine-functionalized polystyrene resin supported Pd(II) complex under aerobic conditions

A polymer-supported palladium(II) N,N-bis(naphthylideneimino)diethylenetriamine complex is found to be a highly active catalyst for Sonogashira coupling reactions. The reactions are performed under copper- and phosphine-free conditions in an air atmosphere. The palladium catalyst is easily separated, and can be reused several times without significant loss in catalytic activity.     

Microfluidic tool based on the antibody-modified paramagnetic particles for detection of 8-hydroxy-2'-deoxyguanosine in urine of prostate cancer patients

Guanosine derivatives are important for diagnosis of oxidative DNA damage including 8-hydroxy-2'-deoxyguanosine (8-OHdG) as one of the most abundant products of DNA oxidation. This compound is commonly determined in urine, which makes 8-OHdG a good non-invasive marker of oxidation stress. In this study, we optimized and tested the isolation of 8-OHdG from biological matrix by using paramagnetic particles with an antibody-modified surface. 8-OHdG was determined using 1-naphthol generated by alkaline phosphatase conjugated with the secondary antibody. 1-Naphthol was determined by stopped flow injection analysis (SFIA) with electrochemical detector using a glassy carbon working electrode and by stationary electrochemical detection using linear sweep voltammetry. A special modular electrochemical SFIA system which needs only 10 μL of sample including working buffer for one analysis was completely designed and successfully verified. The recoveries in different matrices and analyte concentration were estimated. Detection limit (3 S/N) was estimated as 5 pg/mL of 8-OHdG. This method promises to be very easily modified to microfluidic systems as "lab on valve". The optimized method had sufficient selectivity and thus could be used for determination of 8-OHDG in human urine and therefore for estimation of oxidative DNA damage as a result of oxidation stress in prostate cancer patients.