Identification of Escherichia coli by detection of hydroquinone and uracil in the urine system

For rapid identification of Escherichia coli, changes of urinary metabolites incubated with E. coli were investigated by gas chromatography--mass spectrometry. Hydroquinone and uracil were detected and the normal urinary constituent 4-deoxythreonic acid was found to diminish in urine incubated with E. coli. Hydroquinone could not be detected in urine incubated with Klebsiella pneumoniae, Serratia marcescens or Pseudomonas aeruginosa. Although uracil was detected in normal urine, urine incubated with E. coli showed an increased uracil level. Urine incubated with K. pneumoniae, S. marcescens or P. aeruginosa evidenced no such change. A decrease of 4-deoxythreonic acid was noted in urine incubated with S. marcescens or P. aeruginosa. In 7.0 X 10(7) cells of E. coli, 0.33-2.36 micrograms of hydroquinone and 13.4-42.0 micrograms of uracil were detected after 3 h of incubation at 38 degrees C, and production was not changed after 4, 5 or 8 h of incubation. These results suggest that the detection of hydroquinone and uracil in urine is useful for rapid identification of E. coli.     

Stereoselective synthesis of dinucleoside phosphate aryl esters by using new condensing reagents,arenesulfonyl 5-(pyridin-2-yl)-tetrazole1

Deoxyadenylyl-(3'-5')-phosphoro-o-chlorophenyl deoxyadenosine having the Sp configuration was synthesised stereoselectively by condensing N,5'-bis-dimethoxytrityldeoxyadenosine 3'-(o-chlorophenyl) phosphate with N,3'-bis-dimethoxytrityldeoxyadenosine using 1-2,4,6-triisopropylbenzene-5-(pyridin-2-yl) tetrazole as the activating reagent followed by dedimethoxytritylation. The absolute configuration of this diastereomer was determined spectroscopically by comparison of two corresponding diastereomers which were prepared by using a conventional condensing reagent and isolated by chromatography on silica gel. Their nuclear Overhauser effect in proton magnetic resonances and the circular dichroism of these two diastereoisomers could only be explained if the product obtained by the stereoselective synthesis had Sp configuration. Our results also suggest that phosphotriesters of this type exist in solution as an equilibrium of folded and extended forms.     

Doubly-Colored Graphs as Graphical Models for Subductions of Coset Representations, Double Cosets, and Unit Subduced Cycle Indices

The concept of doubly-colored graphs is proposed to model subductions of coset representations, double cosets, and unit subduced cycle indices, which have been mathematically formulated in coset algebraic theory developed by Fujita (Symmetry and Combinatorial Enumeration in Chemistry, Springer-Verlag, Berlin, 1991).     

Availability of 6-hydroxynicotinic acid for rapid identification of Pseudomonas aeruginosa and Serratia marcescens

Gas chromatography--mass spectrometry has been used to identify specific metabolites produced by Gram-negative bacteria such as Pseudomonas aeruginosa, Serratia marcescens, Klebsiella pneumoniae and Escherichia coli in a defined medium. 6-Hydroxynicotinic acid was detected in spent culture media of Pseudomonas aeruginosa and Serratia marcescens, but could not be detected in those of Klebsiella pneumoniae and Escherichia coli. The production of 6-hydroxynicotinic acid was recognized by the addition of nicotinic acid in urine with Pseudomonas aeruginosa or Serratia marcescens, but not without the addition of nicotinic acid. Among 10(5) Pseudomonas aeruginosa per 1 ml of urine (criteria for the diagnosis of urinary tract infection), 0.15 microgram of 6-hydroxynicotinic acid was detected in urine at 4 h incubation with nicotinic acid at the optimum pH of 6.9, 38 degrees C. The production of 6-hydroxynicotinic acid was proportional to the number of the bacteria and displayed a time dependency. These results suggest that the availability of 6-hydroxynicotinic acid might make for more rapid identification of bacteria than current methods.     

Near-infrared Light Emission from Nanomolar-level Singlet Molecular Oxygen Generated with an Ultra low Concentration Mixture of Hypochlorite and Hydrogen Peroxide

Abstract— We report the detection of a weak near-infrared light emission originating from 8 nM singlet molecular oxygen (¹O₂) produced in a mixture of 1 m M hypochlorite (OC1^-) and 8 n M hydrogen peroxide (H₂O₂). The measurements were made with a highly sensitive detection system for ultraweak light emission in the 1.0-1.5 μm wavelength region. The emission intensity exhibited linear dependence for H₂O₂ concentrations in the range of 8-670 n M . The mixture containing a lower concentration (33 μ M ) of OCl^- pseudocontinuously emitted near-infrared light for 5 s. The rate constant for ¹O₂ production obtained from the kinetic analysis agrees with that previously reported. Our results demonstrate the possibility of measuring very low concentrations of ¹O₂ in a OCi-/H₂O² mixture as well as ¹O₂ production in intact living systems.     

Photosensitized oxygenation of sulfides within an amphiphilic dendrimer containing a benzophenone core

Photosensitized oxygenation of sulfides within amphiphilic dendrimers, Gn [n(generation) = 1-3], consisting of a benzophenone (BZP) sensitizing core, apolar interior based on n-undecane spacer, and polar dendron exterior based on 2,2-bis(hydroxymethyl)propionic acid, has been investigated in O(2)-saturated methanol. Sulfoxide formation occurring via reaction of O(2) with triplet excited-state sulfide ((3)sulfide), which is formed by a triplet energy transfer (TET) from photoformed (3)BZP to sulfide, was accelerated by the dendric sensitizers, where G2 showed the highest yields of alkylaryl and dialkyl sulfoxides. Laser photolysis studies revealed that enhanced access of sulfide to the (3)BZP core inside the apolar microenvironment accelerates the TET to sulfide, whereas prompt migration of polar sulfoxide to the polar outer shell of the dendrimer suppresses a competitive TET to sulfoxide, thus resulting in effective (3)sulfide formation. Another notable feature of the dendric sensitizer appears in oxygenation of diaryl sulfide, which is promoted by a persulfoxide intermediate formed by photooxygenation of dialkyl sulfide; photoirradiation of a mixture of diethyl sulfide (1a) and diphenyl sulfide (4a) with G2 gave 17-fold higher diphenyl sulfoxide (4b) yield than that obtained with unmodified BZP. The apolar microenvironment within the dendric sensitizer encapsulates a large quantity of 4a, which is oxidized effectively by the persulfoxide of 1a, thus resulting in high 4b yield. The BZP core within the dendric sensitizer is stable even by photoexcitation in protic solvent, suggesting potential utilities of this dendric system for effective and selective photosensitized oxygenation of sulfides.     

Quantitation of formate by solid-phase microextraction and gas chromatography--mass spectrometry utilizing a [13C]formate internal standard

A new method for the analysis of formic acid was developed using gas chromatography-electron impact ionization mass spectrometry in the selected ion monitoring mode and solid-phase microextraction. Using this method with [13C]formic acid as an internal standard, the peak area ratio of [12C]formic acid/[13C]formic acid was not affected by differing methanol or sulfuric acid concentrations during the esterification and fiber adsorbing step. In comparison, the peak area ratio of formic acid/acetonitrile as detected by conventional GC with flame ionization detection was greatly affected by methanol or sulfuric acid concentrations. The formic acid calibration curve of our method showed excellent linearity over the range 5 to 200 microM. The within- and between-run assay relative standard deviations for the formic acid concentration were all less than 1.70%.     

Facile fabrication of Ag-Pd bimetallic nanoparticles in ultrathin TiO(2)-gel films: nanoparticle morphology and catalytic activity

Ag-Pd bimetallic nanoparticles were prepared directly in ultrathin TiO(2)-gel films by a stepwise ion-exchange/reduction approach. Ion-exchange sites were created in ultrathin films using Mg(2+) ions as template. Ag(+) ion was then incorporated by ion exchange, and converted into metallic nanoparticles by low-temperature H(2) plasma, regenerating ion-exchange sites. The same procedure was then carried out for Pd(2+) ion, producing Pd-on-Ag bimetallic nanoparticles, as TEM observation and plasmon resonance absorption indicate. By contrast, reversed metal incorporation procedure appeared to give a mixture of individual Ag and Pd nanoparticles, as confirmed by TEM, absorption spectroscopy and X-ray photoelectron spectroscopy. For hydrogenation of methyl acrylate, the catalytic activity of the Pd-on-Ag nanoparticle is 367 times as large as that of commercial Pd black and 1.6 times as large as that of Pd monometallic nanoparticle. The outstanding catalytic activity was explicable by the large fraction of the surface-exposed Pd atoms. The formation process of the bimetallic nanoparticle and their general morphological feature are discussed.     

Evaluation of diastolic phase by left ventricular volume curve using S2-gated equilibrium method in radioisotope angiography

S2-gated (the second heart sound) method was designed by authors. In 6 normal subjects and 16 patients (old myocardial infarction 12 cases, hypertension 2 cases and aortic regurgitation 2 cases), radioisotope (RI) angiography using S2-gated equilibrium method was performed. In RI angiography, 99mTc-human serum albumin (HSA) 555MBq (15 mCi) as tracer, PDP11/34 as minicomputer and PCG/ECG synchronizer (Metro Inst.) were used. Then left ventricular (LV) volume curve by S2-gated and electrocardiogram (ECG) R wave-gated method were obtained. Using LV volume curve, left ventricular ejection fraction (EF), mean ejection rate (mER, s-1), mean filling rate (mFR, s-1) and rapid filling fraction (RFF) were calculated. mFR indicated mean filling rate during rapid filling phase. RFF was defined as the filling fraction during rapid filling phase among stroke volume. S2-gated method was reliable in evaluation of early diastolic phase, compared with ECG-gated method. There was the difference between RFF in normal group and myocardial infarction (MI) group (p less than 0.005). RFF in 2 groups were correlated with EF (r = 0.82, p less than 0.01). RFF was useful in evaluating MI cases who had normal EF values. The comparison with mER by ECG-gated and mFR by S2-gated was useful in evaluating MI cases who had normal mER values. mFR was remarkably lower than mER in MI group, but was equal to mER in normal group approximately. In conclusion, the evaluation using RFF and mFR by S2-gated method was useful in MI cases who had normal systolic phase indices.     

Synthesis of a new chiral source, (1R,2S)-1-Phenylphospholane-2-carboxylic acid, via a key intermediate alpha-phenylphospholanyllithium borane complex: configurational stability and X-ray crystal structure of an alpha-monophosphinoalkyllithium borane complex

A synthetic route to enantiomerically pure (1R,2S)-1-phenylphospholane-2-carboxylic acid (1), which is a phosphorus analogue of proline, has been established. A key step is the deprotonation-carboxylation of the 1-phenylphospholane borane complex 3 by using sBuLi/1,2-dipiperidinoethane (DPE). Configurational stability of the key intermediate, the amine-coordinated alpha-phosphinoalkyllithium borane complex 4, was investigated by employing lithiodestannylation-carboxylation of both diastereomers of the 1-phenyl-2-trimethylstannylphospholane borane complex 7 in the presence of several kinds of amines, and as a result, 4 was found to be configurationally labile even at -100 degrees C. The key intermediate, the DPE-coordinated trans-1-phenyl-2-phospholanyllithium borane complex 9, was isolated, and the structure was identified by X-ray crystal structure analysis. This is the first X-ray crystal structure determined for an alpha-monophosphinoalkyllithium borane complex. Remarkably, the alkyllithium complex is monomeric and tricoordinate at the lithium center with a slightly pyramidalized environment, and the existence of a Li--C bond (2.170 A) has been confirmed. Moreover, (1)H-(7)Li HOESY and (6)Li NMR analyses suggested the structure of 9 in solution as well as the existence of an equilibrium between 9, its cis isomer, and the ion pair 8 at room temperature, which was extremely biased towards 9 at -100 degrees C. Finally, 1 was used as a chiral ligand in a palladium-catalyzed allylic substitution, and the desired product was obtained in high yield with good enantioselectivity.