Dry rehydratable film method for rapid enumeration of coliforms in foods (3M Petrifilm Rapid Coliform Count plate): collaborative study

A rehydratable dry-film plating method for coliforms in foods, the 3M Petrifilm Rapid Coliform Count plate method, was compared with the U.S. Food and Drug Administration's Bacteriological Analytical Manual method for nondairy foods and the American Public Health Association's Standard Methods for the Examination of Dairy Products (SMEDP) method for dairy foods. Six food types, vanilla ice cream, cheddar cheese, fresh refrigerated uncooked pasta, wheat flour, prepared frozen macaroni and cheese, and frozen hash browns, were analyzed for coliforms by 11 collaborating laboratories. For each food product tested, the collaborators received 8 blind samples consisting of a control sample and 3 levels of inoculated sample, each in duplicate. The mean log counts for the methods were comparable. The repeatability and reproducibility variances of the Petrifilm Rapid Coliform Count method at 14 and 24 h were not significantly different from those of the standard methods.     

Restricted access chromatographic sample preparation of low mass proteins expressed in human fibroblast cells for proteomics analysis

Two-dimensional electrophoresis and modern image analysis systems have made it possible to study protein expression and regulation of proteins in biological systems. Proteins in the molecular mass region of 20-120 kDa are well investigated and described. However, proteins with masses below 20 kDa are the least investigated as they are rarely seen on 2D-PAGE due to fast migrations in the electric field and lack of staining efficiency. This paper describes a technique that enriches proteins in the lower mass region using solid-phase extraction. The purification step is carried out using C18 functionalised "restricted access" affinity chromatography whereby simultaneous trace enrichment and sample clean up is achieved. In this study expression patterns of TGF-beta stimulated and non-stimulated fibroblasts were compared after the solid-phase fractionation procedure. An increased expression pattern was obtained whereby 400 protein spots could be detected by image analysis in the <20-kDa region. Out of these, specific regulations of 14 spots were found by quantitative image analysis and spots of interest were identified with MALDI TOF-MS. The regulated and identified proteins were triosephosphate isomerase, cofilin and heat shock 27-kDa protein.     

Selective and sensitive high-performance liquid chromatographic assay for the metabolites of nomifensine in human plasma

A selective high-performance liquid chromatographic method for the determination of the three metabolites of nomifensine in human plasma is described. All metabolites and the internal standard, mexiletine, are extracted with diethyl ether and then back-extracted into an acidic aqueous phase. After subsequent extraction into diethyl ether the metabolites are analysed by high-performance liquid chromatography. A reversed-phase C18 column is used with a mobile phase of dioxane-methanol-potassium phosphate buffer (pH 2.25). The sensitivity of the method is 0.007 micromol/l for all metabolites. Extraction efficiencies are 84.6%, 75.8%, and 78.2% for 4'-hydroxynomifensine, 4'-hydroxy-3'-methoxynomifensine and 3'-hydroxy-4'-methoxynomifensine, respectively. The reproducibility of the method is good, the coefficients of variation (%) varying between 2.1% and 9.9% in the concentration range 0.05-1.00 micromol/l. The procedure was applied to human plasma samples from a volunteer who had received a single oral dose of nomifensine. The method is accurate and sensitive for pharmacokinetic studies on the metabolites of nomifensine.     

3M Petrifilm enterobacteriaceae count plate method for enumeration of enterobacteriaceae in selected foods: collaborative study

The practice of detecting and enumerating all oxidase-negative, glucose-fermenting-Gram-negative rods (i.e., the family Enterobacteriaceae) is used to indicate unsanitary or inadequate food processing conditions. The objective of this interlaboratory collaborative study was to evaluate and compare the methods described in Standard Methods for the Examination of Dairy Products (SMEDP) and the Compendium of Methods for the Microbiological Examination of Foods (Compendium) with a commercial product, the 3M Petrifilm Enterobacteriaceae Count Plate, for the recovery of Enterobacteriaceae in foods. Six foods--cheddar cheese, milk, flour, frozen prepared meals, frozen broccoli, and nut pieces--were analyzed for Enterobacteriaceae by 12 collaborating laboratories. For each food tested, the collaborators received 8 blind test portions consisting of a control test portion and 3 levels of inoculated test portion, each in duplicate. Each test portion was tested by the Petrifilm Enterobacteriaceae Count Plate method as well as the SMEDP or Compendium methods. The precision estimates (repeatability or within-laboratory variation, and reproducibility or between-laboratory variation) were calculated with standard statistical techniques.     

Nonlinear forward scattering of resonant laser light from excited neon systems

We investigate forward scattering induced by a powerful single mode dye laser interacting with excited neon systems. We present experimental data obtained with the three transitions: 1s ₄(J=1)-2p ₃(J=0), 1s ₂(J=1)-2p ₁(J=0), and 1s ₅(J=2)-2p ₄(J=2). The lineshape dependences on the gas pressure, the laser power and the laser field fluctuations are discussed in the context of published theories. Our data resemble the calculated lineshapes in which fluctuations in the interaction processes between atoms and laser fields are taken into account. In atomic systems having more complex level degeneracies [1s ₅(J=2)-2p ₄(J=2)] signal structures expected from higher order nonlinear coherences can occur. To theoretically reproduce these lineshapes a simple J=1J=0 model is fully appropriate provided that the level with J=1 has a narrow width.     

Action spectra for survival and spore photoproduct formation of Bacillus subtilis irradiated with short-wavelength (200-300 nm) UV at atmospheric pressure and in vacuo.

Spores of Bacillus subtilis are approximately ten times less likely to survive UV light irradiation in a vacuum than under atmospheric conditions. Photoproduct formation was studied in spores irradiated under ultrahigh vacuum (UHV) conditions and in spores irradiated at atmospheric pressure. In addition to the "spore photoproduct" 5-thyminyl-5,6-dihydrothymine (TDHT), which is produced in response to irradiation at atmospheric pressure, two additional photoproducts, known as the cis-syn and trans-syn isomers of thymine dimer, are produced on irradiation in vacuo. The spectral efficiencies for photoproduct formation in spores are reduced under vacuum conditions compared with atmospheric conditions by a factor of 2-6, depending on the wavelength. Because formation of TDHT does not increase after irradiation in vacuo, TDHT cannot be responsible for the observed vacuum effect. Vacuum specific photoproducts may cause a synergistic response of spores to the simultaneous action of UV light and UHV. An increased quantum efficiency, destruction of repair systems and formation of irreparable lesions are postulated for the enhanced sensitivity of B. subtilis spores to UV radiation in vacuo.     

Tunable feedback loop for suppression of relaxation oscillations in a diode-pumped Nd: laser

An optoelectronic noise suppression circuit connected to the RF input of a commercial diode laser mount is presented. Adjustable phase shift and amplitude control enable efficient suppression of the relaxation oscillations of a diode laser-pumped Nd:YVO₄ laser at variable frequencies between a few hundred kHz and 1.8 MHz. At least 25 dB reduction of the relaxation oscillations at 500 kHz and 1.5 MHz noise peaks is demonstrated.     

Pionic atom factory project in the RIBF

We are preparing for a high resolution spectroscopy of the 1s and 2s pionic states of ¹²¹Sn in the ¹²²Sn(d,³He) reaction at the incident deuteron energy of ~500 MeV. The measurement will provide accurate information on the strong interaction between the pion and the nucleus, which leads to quantitative determination of the magnitude of the quark condensate at the normal nuclear density.