Rapid identification of Escherichia coli and Helicobacter pylori in biological samples by capillary zone electrophoresis

The differences in surface charge of different bacteria can be exploited for their separation by capillary electrophoresis. However, this method of separation of microorganisms is beset with various drawbacks such as adhesion of bacteria to the fused silica surface or cluster formation. To overcome these phenomena we investigated the addition of poly(ethylene oxide) as a focusing agent to the running buffer and used calcium and myoinositol hexakisphosphate as specific ions that interact with the bacterial surface, changing its electrical properties and electrophoretic mobilities. In the present work, we applied CZE to identification of E. coli in infected urine (direct injection) from patients with urinary tract infections and to identification of Helicobacter pylori, which is a gram-negative bacillus responsible for one of the most common infections found in humans worldwide. Helicobacter pylori colonize the stomach and are responsible for severe diseases of the gastric tract, ranging from chronic gastric ulcer to gastric cancer.     

Selective and reversible control of a chemical reaction with narrow-band infrared radiation: HXeCC radical in solid xenon

The light-induced H + XeC2 <--> HXeCC reaction is studied in solid Xe, and the full optical control of this reaction is demonstrated. By narrow-band excitation in the IR spectral region, HXeCC radicals can be decomposed to a local metastable configuration and then selectively recovered by resonant excitation of the XeC2 vibrations. The novel recovery process is explained by short-range mobility of the reagents promoted by vibrational energy redistribution near the absorbing XeC2 molecule. This means that a chemical reaction can be selectively promoted in a desired place where the chosen absorber locates. The obtained results make a strong case of solid-state reactive vibrational excitation spectroscopy of weak radiationless transitions.     

Characterization of nodularin variants in Nodularia spumigena from the Baltic Sea using liquid chromatography/mass spectrometry/mass spectrometry

Nodularin is a potent hepatotoxic cyclic pentapeptide produced by planktonic cyanobacterium Nodularia spumigena. Bloom and culture samples of the cyanobacterium collected and isolated from the Gulf of Gdańsk, southern Baltic Sea, were analyzed. Hybrid quadrupole-time-of-flight liquid chromatography/mass spectrometry/mass spectrometry (TOF-LC/MS/MS) with ionspray (ISP) and collision-induced dissociation (CID) were used to characterize nodularin and its analogues. The identification process was based on the comparison of recorded product ion spectra with the previously reported FAB-MS/CID (high-energy) mass spectra of the corresponding nodularin variants. Amino acid structures and sequences were derived from the fragmentation pattern of the [M+H](+) ions. Apart from unmodified nodularin with an arginine residue (NOD-R), three demethylated variants have been found. The sites of demethylation were located on aspartic acid [Asp(1)]NOD, the Adda residue [DMAdda(3)]NOD, and dehydrobutyric acid [dhb(5)]NOD. In two other nodularin variants an additional methyl group is located in the Adda [MeAdda]NOD and Glu [Glu(4)(OMe)]NOD residues. The linear NOD and the geometrical isomer of NOD-R, reported earlier in N. spumigena from New Zealand, have also been detected. Two of the total eight nodularin variants characterized in the present study, [dhb(5)]NOD and [MeAdda]NOD, have not been described earlier.     

A comparison of rigid and flexible water models in collisions of monomers and small clusters

In this study we have investigated the dynamics of small water clusters using microcanonical molecular dynamics simulations. The clusters are formed by colliding vapor monomers with target clusters of two and five molecules. The monomers are sampled from a thermal ensemble at T=300 K and target clusters with several total energies are considered. We compare rigid extended simple point charge water with flexible counterparts having intramolecular harmonic bonds with force constants 10(3) and 10(5) kcal(mol A2). We show that the lifetimes of the clusters formed via collision process are similar for the rigid model and the flexible model with the bigger force constant, if the translational temperatures of the target cluster molecules are equal. The model with the smaller force constant results in much longer lifetimes due to the stabilizing effect caused by the kinetic energy transfer into internal vibration of the molecules. This process may take several hundreds of picoseconds, giving rise to time-dependent decay rates of constant-energy clusters. A study of binary collisions of water molecules shows that the introduction of flexibility to the molecules increases the possibility of dimer formation and thus offers an alternative route for dimer production in vapors. Our results imply that allowing for internal degrees of freedom is likely to enhance gas-liquid nucleation rates in water simulations.     

Unimolecular processes in CH2OH below the dissociation barrier: O-H stretch overtone excitation and dissociation

The OH-stretch overtone spectroscopy and dynamics of the hydroxymethyl radical, CH(2)OH, are reported in the region of the second and third overtones, which is above the thermochemical threshold to dissociation to H+CH(2)O (D(0)=9600 cm(-1)). The second overtone spectrum at 10 484 cm(-1) is obtained by double resonance IR-UV resonance enhanced multiphoton ionization (REMPI) spectroscopy via the 3p(z) electronic state. It is rotationally resolved with a linewidth of 0.4 cm(-1) and displays properties of local-mode vibration. No dissociation products are observed. The third overtone spectra of CH(2)OH and CD(2)OH are observed at approximately 13 600 cm(-1) by monitoring H-atom photofragments while scanning the excitation laser frequency. No double resonance REMPI spectrum is detected, and no D fragments are produced. The spectra of both isotope analogs can be simulated with a linewidth of 1.3 cm(-1), indicating dissociation via tunneling. By treating the tunneling as one dimensional and using the calculated imaginary frequency, the barrier to dissociation is estimated at about 15 200 cm(-1), in good agreement with theoretical estimations. The Birge-Sponer plot is linear for OH-stretch vibrations 1nu(1)-4nu(1), demonstrating behavior of a one-dimensional Morse oscillator. The anharmonicity parameter derived from the plot is similar to the values obtained for other small OH containing molecules. Isomerization to methoxy does not contribute to the predissociation signal and the mechanism appears to be direct O-H fission via tunneling. CH(2)OH presents a unique example in which the reaction coordinate is excited directly and leads to predissociation via tunneling while preserving the local-mode character of the stretch vibration.     

Postcollision relaxation of small atomic clusters

Molecular-dynamics simulations are performed to investigate the effects caused by the lack of internal equilibration on the dynamics and properties of atomic clusters. The studied systems consist of Lennard-Jones clusters of five to ten atoms and a colliding vapor monomer. Cluster radius and potential energy are shown to reach a time-independent value within 30 ps after a collision with a vapor monomer. The relaxation in terms of rotational energy takes at least 200 ps. During the first couple of picoseconds after the collision time-dependent cluster decay rates are observed. The unrelaxed cluster states are expected to have minimal effect on gas-liquid nucleation rates.     

The synthesis of opiorphin and studies on its binding ability toward Cu(II)

Opiorphin (OPI) is an endogenous pentapeptide isolated from human saliva. A characteristic feature of this peptide is the presence of an N-terminal Gln residue. The synthesis and binding abilities of opiorphin toward Cu(II) ions are described. The use of potentiometric and spectroscopic methods allows us to propose the binding mode of the formed complexes.     

UV-Vis Investigation on the Hydrocarbon Length Effect on the α-Tocopherol Solubility in an AOT/n-Alkane/Water System

The influence of n-alkane hydrocarbon chain length on both binding and distribution constants of α-tocopherol and Aerosol-OT [sodium bis(2-ethylhexyl) sulfosuccinate] reversed micelles were studied with the UV-vis method using n-heptane and n-decane as solvents. The amount of water in the system was determined by R defined as the ratio of water to surfactant molalities ( $R=[\hbox{H$_{2}$O}]/[\hbox{AOT}]$ ). No significant water influence on distribution and binding constants was observed. This finding is consistent with earlier results, which indicated that the location of α-tocopherol molecules in the AOT reversed micelles is the palisade layer. The results obtained indicate that the longer hydrocarbon chain length makes surroundings of micelles more ordered and causes some limitations in α-tocopherol access to its palisade layer. This implies a weaker connection to the micellar structure and an increase of α-tocopherol freedom of motion in the space of hydrocarbon tails around polar core.     

Quantitative analysis of phosphatidylcholines and phosphatidylethanolamines in urine of patients with breast cancer by nanoflow liquid chromatography/tandem mass spectrometry

Phosphatidylcholines (PCs) and phosphatidylethanolamines (PEs) from urine of patients with breast cancer were qualitatively and quantitatively analyzed by nanoflow liquid chromatography-electrospray ionization tandem mass spectrometry (nLC-ESI-MS-MS). Urinary phospholipids (PLs) were extracted from three different categories of patients (non-cancer controls and breast cancer patients before and after surgery) by first lyophilizing only 1 mL of urine sample to enrich PLs. Next, nLC-ESI-MS-MS analysis of intact urinary phospholipids was performed, resulting in structural identification of 21 PCs and 12 PEs, followed by quantitative analysis using a multiple standard addition method. This study demonstrated that nLC-ESI-MS-MS can be powerfully utilized for the study of relative changes in the contents and concentration of urinary PCs and PEs from breast cancer patients: total concentration of PCs and PEs of patient sample increased to (144?±?9)% and (171?±?11)%, respectively, compared to control sample but they decreased significantly following surgery.