Sugar sweetness and pleasantness: evidence for different psychological laws.

Sweetness and the pleasantness of sweetness of sucrose solutions and sweetened food conform to different functions. Sweetness rises with concentration, whereas pleasantness first rises and then decreases. The breakpoint appears to occur at a constant sweetness (that is, constant sensory) level.

Purification and properties of methanol dehydrogenase from Hyphomicrobium x.

(1) A method for the isolation of methanol dehydrogenase (alcohol:(acceptor) oxidoreductase, EC 1.1.99.8) from Hyphomicrobium X is decribed. The purified enzyme was resolved by polyacrylamide gel electrophoresis into one main and two minor active bands. Iron and manganese were the only detected metals in the enzyme preparation. (2) The substrate, methanol, was oxidized to formic acid by a stoichiometric amount of artificial electron acceptor. During the reaction, no free formaldehyde could be detected. Other primary alcohols were oxidized to the corresponding aldehydes were a poor substrate or no substrate at all. (3) Some new and efficient one-electron acceptors were found. With these electron acceptors, the enzyme had a high pH optimum and ammonia was still required in the assay system. (4) ESR spectroscopy showed the presence of an enzyme-bound organic free radical. With X-band ESR the signal had a peak-to-peak linewidth of about 0.7 mT. The signal was further resolved by Q-band ESR and the values gparallel = 2.0024 and gperpendicular = 2.0056 were derived. (5) Under denaturing conditions the ESR signal and enzymatic activity disappeared at the same time as fluorescence appeared. Enzymatic activity is not restored when extracted cofactor and apoenzyme are brought together under normal conditions. Some properties of the unusual prosthetic group are presented in a preliminary form.

Optimum recovery of Mycobacterium avium complex from blood specimens of human immunodeficiency virus-positive patients by using small volumes of isolator concentrate inoculated into BACTEC 12B bottles.

Recovery of Mycobacterium avium complex organisms from 412 lysis-centrifugation (Isolator) concentrates of blood specimens obtained from human immunodeficiency virus-positive individuals was attempted with the following media (and Isolator concentrate inoculum volumes): BACTEC 12B broth (0.2 and 1.0 ml), Lowenstein-Jensen slants (0.1 ml), and Middlebrook 7H10/11 agar (0.1 ml). A total of 42 M. avium complex isolates were recovered. The highest rates of recovery and shortest detection times were noted with BACTEC 12B bottles inoculated with 0.2 ml of Isolator concentrate. Middlebrook agar was superior to Lowenstein-Jensen slants. Fluorochrome acid-fast smears performed directly upon Isolator concentrates were of no utility.

Covalent addition of H2O, enzyme substrates and activators to pyrrolo-quinoline quinone, the coenzyme of quinoproteins.

The fluorescence excitation and the adsorption spectrum of 2,7,9-tricarboxy-1H-pyrrolo [2,3-f]quinoline-4,5-dione (pyrrolo-quinoline quinone, PQQ), measured in the pH range 7.0-10.0, are quite different. However, when the temperature of the solution is lowered, the shape and maxima of these spectra become more similar. 1H-NMR in 2H2O revealed a temperature-dependent equilibrium between PQQ and a hydrated form. Evidence is presented that low temperature favours the formation of the fluorescing species which is PQQ, hydrated at the C-5 position. Even further hydration is possible since absorption, fluorescence and NMR spectroscopy of PQQ in borate buffer pH 10.0 reveal additional hydration at the C-4 position, pointing to a dihydrate. PQQ also reacts with quinoprotein enzyme substrates and activators. Spectroscopic measurements showed the existence of 5-alkoxy-5-hydroxy-PQQ and 5-amino-5-hydroxy-PQQ in the presence of alcohols and 2 M NH4Cl, pH 9.0, respectively. In the latter case, the existence of 5-imino-PQQ could also be demonstrated. Addition compounds with amines appear to be unstable. The amines become probably oxidized because pyrrolo-quinoline quinol (PQQH2) was found as the reaction product. On the other hand, an addition compound containing an imino bond could be isolated after addition of urea to a PQQ solution. Spectral characteristics of PQQ and its addition compounds are presented since these data are necessary for the spectral analysis of quinoproteins and the quantitative estimation of coenzyme.

Long-term laboratory contamination by Mycobacterium abscessus resulting in two pseudo-outbreaks: recognition with use of random amplified polymorphic DNA (RAPD) polymerase chain reaction.

Beginning in 1993, an increase in clinical isolates of Mycobacterium abscessus was observed in a single hospital microbiology laboratory. This involved a cluster of four patients in June 1993 and five patients and a quality-control culture of distilled water in May 1994. Twenty-three M. abscessus isolates recovered between 1991 and 1996 were compared by random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). Sixteen of 21 clinical isolates recovered over a 6-year period and the distilled water isolate had identical RAPD-PCR patterns consistent with a single strain or clone. Only six of 15 patients had findings suggestive of clinical disease. Since the use of in-house-prepared distilled water was discontinued, no further laboratory contamination of clinical specimens has been observed. Molecular typing was the key to defining distilled water as the source of this pseudo-outbreak. Recognition of such outbreaks is important for prevention of unnecessary therapeutic and diagnostic interventions.