Analysis of breath by proton transfer reaction time of flight mass spectrometry in rats with steatohepatitis induced by high-fat diet

Breath testing has been largely used as a diagnostic tool, but the difficulties in data interpretation and sample collection have limited its application. We developed a fast (< 20?s), on-line, non-invasive method for the collection and analysis of exhaled breath in awake rats based on proton transfer reaction time of flight mass spectrometry (PTR-ToF-MS) and applied it to investigate possible relationships between pathologies induced by dietary regime and breath composition. As a case study, we investigated rats with dietary induced non-alcoholic steatohepatitis (NASH) and modifications induced by coffee addition to the diet. We considered two different diets (standard and high fat) complemented with two different drinking possibilities (water or decaffeinated coffee) for a total of four groups with four rats each. Several spectrometric peaks were reliable markers for both dietary fat content and coffee supplementation. The high resolution and accuracy of PTR-ToF-MS allowed the identification of related compounds such as methanol, dimethyl sulphide, dimethyl sulphone and ammonia. In conclusion, the rapid and minimally invasive breath analysis of awake rats permitted the identification of markers related to diet and specific pathologic conditions and provided a useful tool for broader metabolic investigations. Copyright ? 2012 John Wiley & Sons, Ltd.     

Identification of glucosinolates in capers by LC-ESI-hybrid linear ion trap with Fourier transform ion cyclotron resonance mass spectrometry (LC-ESI-LTQ-FTICR MS) and infrared multiphoton dissociation

An liquid chromatography-mass spectrometry method using electrospray ionization in negative ion mode coupled with a hybrid quadrupole linear ion trap and Fourier transform ion cyclotron resonance (FTICR) mass spectrometer was applied to characterize of intact glucosinolates (GLSs) in crude sample extracts of wild bud flowers of Capparis spinosa (Capparis species, family Capparaceae). Structural information of GLSs was obtained upon precursor ions' isolation within the FTICR trapping cell and subsequent fragmentation induced by infrared multiphoton dissociation (IRMPD). Such a fragmentation was found very useful in terms of chemical identification of all precursor ions [M-H](-) including sulfur-rich GLSs reported here for the first time. Along with most common GLSs already found in capers such as glucocapparin, isopropyl/n-propyl-GLS, mercapto-glucocapparin, and two indolic GLS, i.e., 4-hydroxyglucobrassicin and glucobrassicin, the occurrence of the uncommon glycinyl-glucocapparin as well as two sulfur-rich GLSs is reported. IRMPD showed an increased selectivity towards disulfide bond cleavages with thiol migration, suggesting the side chain structure of non-targeted compounds, i.e., disulfanyl-glucocapparin and trisulfanyl-glucocapparin. Glucocapparin [2.05?±?0.25?mg/g, dry weight (dw)] was the most abundant GLS, followed by glucobrassicin (232?±?18?μg/g, dw) and 4-hydroxyglucobrassicin (89?±?12?μg/g, dw). All other compounds were present at very low content ranging from 0.5 to 1.5?μg/g dw. Copyright ? 2012 John Wiley & Sons, Ltd.     

The Use of DRAQ5 to Monitor Intracellular DNA in <Emphasis Type="Italic">Escherichia coli</Emphasis> by Flow Cytometry

Flow cytometry provides a rapid and high-content multiparameter analysis of individual microorganisms within a population. In the past years, several fluorescent stains were developed in order to monitor DNA content distribution and cell-cycle phases, mainly in eukaryotic cells. Recently, due to its low detection limits, several of these fluorescent stains were also applied to prokaryotic cells. In this study, the ability of a novel far-red fluorescent stain DRAQ5 in assessing intracellular DNA content distribution in Escherichia coli DH5α was evaluated. The results showed that a DRAQ5-labelled live E. coli suspension can be obtained by incubation of 1 × 10⁶ cells/mL with 5 μM DRAQ5 in PBS buffer supplemented with EDTA (pH = 7.4) during 30 min at 37 °C. Flow cytometric analysis of fixed E. coli cells revealed that ethanol should be used in detriment of glutaraldehyde for DRAQ5 labelling. After the analysis of RNase and DNase digested samples, DRAQ5 was proven to be a specific DNA labelling stain. The present study demonstrates that the use of DRAQ5 as a DNA-labelling stain provides an easy assessment of intracellular DNA content and cell-cycle phases in Gram-negative bacteria such as E. coli.     

Reassessment of pH reference values with improved methodology for the evaluation of ionic strength

The conflict between pH as empirical number in routine control and the pH value regarded as conveying some information concerning the effective concentration or activity of hydrogen ions, a_H, has caused much confusion.There are, however, reasons to conclude that the overwhelming amount of thermodynamic data is not sufficiently accurate—either due to ignorance of metrological concepts or due to insufficiently specified measurement processes of fundamental chemical quantities pH.The commonly used seven reference buffer solutions to which primary pH values have been conventional assigned, represent a selection out of a more extensive list, recommended by NBS (now NIST) in 1962. From then onwards conventions concerning the Debye-Hückel model of electrolyte solutions and ionic strength have been revised and the pH(S) values reassessed in conformity but only for these seven reference buffer solutions. The others have, so far remained unchanged, locking harmonisation of the conventionally assigned pH(S) values.In this work, ionic strength is calculated through complete equations derived from the acidity constants. Concentrations of the various species involved in the conventional assignment of pH and their corresponding activity coefficients are therefore, more rigorously known. The process proves particularly useful for poliprotic acids with overlapping acidity constants, where the ratio is less than 10³.As a consequence, conventionally assigned pH values of reference buffer solutions are recalculated and corrections are introduced as appropriate.     

Solution enthalpies of 1-bromoadamantane in monoalcohols at 298.15 K

Solution enthalpies of 1-bromoadamantane in 14 monoalcohols were measured at 298.15 K. Comparison with the solution enthalpies of 2-bromo-2-methylpropane was performed. The obtained data were discussed in terms of the effect of solute's size and solvent's structure.     

Simultaneous N2O and NO reduction over carbon supported catalysts

The simultaneous reduction of N₂O+NO with carbon supported Ni/K catalysts was studied. High conversions of both compounds were obtained at 400°C, and a synergystic effect between Ni and K was observed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Characterisation of the essential oil of Nepeta glomerata Montbret et Aucher ex Bentham from Lebanon and its biological activities

The essential oil of Nepeta glomerata from Lebanon was studied by means of GC and GC/MS analysis; 70 compounds were identified. The oil was constituted mainly by monoterpenes and the most abundant components were α-pinene, spathulenol and carvacrol. Nepeta glomerata oil showed antibacterial activity, particularly towards Gram-positive bacteria, and also inhibited LPS-induced NO production in macrophage cell line RAW 264.7, with an IC(50) value of 78.1 μg mL(-1). Furthermore, an in?vitro cytotoxic assay showed that the oil was more active on a renal adenocarcinoma cell line (48% of inhibition of proliferation at 100 μg mL(-1)) in comparison to an amelanotic melanoma.     

Solutions of Semilinear Elliptic Equations in Tubes

Given a smooth compact k-dimensional manifold Λ embedded in ? ^m , with m≥2 and 1≤k≤m?1, and given ?>0, we define B _? (Λ) to be the geodesic tubular neighborhood of radius ? about Λ. In this paper, we construct positive solutions of the semilinear elliptic equation $$\begin{cases} \Delta u + u^p = 0 &\mbox{in } B_{\epsilon}(\varLambda) \\ u = 0 & \mbox{on } \partial B_{\epsilon}(\varLambda) , \end{cases} $$ when the parameter ? is chosen small enough. In this equation, the exponent p satisfies either p>1 when n:=m?k≤2 or $p\in(1, \frac{n+2}{n-2})$ when n>2. In particular, p can be critical or supercritical in dimension m≥3. As ? tends to 0, the solutions we construct have Morse index tending to infinity. Moreover, using a Pohozaev type argument, we prove that our result is sharp in the sense that there are no positive solutions for $p>\frac{n+2}{n-2}$ , n≥3, if ? is sufficiently small.     

Sign-changing stationary solutions and blowup for a nonlinear heat equation in dimension two

Consider the nonlinear heat equation $$v_t -\Delta v=|v|^{p-1}v \qquad \qquad \qquad (NLH)$$ in the unit ball of \({\mathbb{R}^2}\) , with Dirichlet boundary condition. Let \({u_{p,\mathcal{K}}}\) be a radially symmetric, sign-changing stationary solution having a fixed number \({\mathcal{K}}\) of nodal regions. We prove that the solution of (NLH) with initial value \({\lambda u_{p,\mathcal{K}}}\) blows up in finite time if |λ ?1| > 0 is sufficiently small and if p is sufficiently large. The proof is based on the analysis of the asymptotic behavior of \({u_{p,\mathcal{K}}}\) and of the linearized operator \({L= -\Delta - p | u_{p,\mathcal{K}} | ^{p-1}}\) .