Involvement of cytochrome P450 in hydroxylation of propylbenzene by Fusarium moniliforme strain MS31

Fusarium moniliforme strain MS31 can oxidize propylbenzene to (R)-1-phenylpropanol with what may be a cytochrome P450. Hydroxylation of propylbenzene needed molecular oxygen, and NADPH as a coenzyme gave a higher yield than NADH. The hydroxylation proceeded further when FAD and FMN were added than in their absence, suggesting that the enzyme was a flavo-protein. Carbon monoxide inhibited the hydroxylation, as did other cytochrome P450 inhibitors such as SKF 525A and miconazole. These characteristics matched those of a microsomal cytochrome P450 monooxygenase system that contained NADPH-cytochrome P450 reductase.     

Screening for microorganisms with specific characteristics by flow cytometry and single-cell sorting

Flow cytometry used in combination with single-cell sorting is a powerful technique for the identification and isolation of microbial cells with particular characteristics, especially when such cells grow more slowly than other cells in a large heterogeneous population. Many applications of flow cytometry with cell sorting, originally used by specialists studying mammalian cells, have been modified so that microorganisms also can be evaluated. The methods can now be used more widely because of the increasing availability of the expensive equipment. There are means for the fluorescence detection of a wide variety of properties, such as amounts of various cell components, specific sequences of peptides and nucleotides, cell functions, and enzyme activities. From the extensive literature, representative reports of an assortment of uses of flow cytometry with cell sorting are reviewed in this article, intended to introduce the technique and its many advantages to microbiologists.     

Molecular Design, Characterization, and Application of Multiinformation Dyes (MIDs) for Optical Chemical Sensings. 3. Application of MIDs for λ(max)-Tunable Ion-Selective Optodes

By utilizing "multiinformation dyes (MIDs)", which have plural spectral change characteristics such as an absorption maximum wavelength (λ(max)) shift based on a polarity change and an absorbance change due to protonation, novel λ(max)-tunable ion-selective optodes were proposed and prepared by employing MIDs with membrane solvents having different polarities. For controlling the detecting λ(max) of the optode, the novel polar membrane solvent [2-[[6-(2-nitrophenoxy)hexyl]oxy]methyl]isobutane-1,3-diol was designed and synthesized, which was used together with a typical membrane solvent nitrophenyl octyl ether. By mixing these two membrane solvents, the λ(max) position of the optode detection wavelength can be shifted and controlled and was successfully applied to a λ(max)-tunable Li(+)-selective optode based on a highly Li(+)-selective ionophore TTD14C4. The λ(max) tuning technique is useful for preparing an optode system using a low-cost light source such as a light-emitting diode or a popular laser.     

Photoautotrophic cultures of the host and transformed cells of Marchantia polymorpha under controlled incident light intensity

Photoautotrophic cultures of the host and transformed cells of the liverwort, Marchantia polymorpha, were examined. In cultures in flat glass flasks under various light intensities, it was found that the growth rates of both the cells increased with increase in light intensity in the range of 0 to 25 W/m2, but further increase in light intensity caused photoinhibition of the growth of the cells. Cultures of both the types of cells under light-controlled conditions using an externally illuminated bioreactor were carried out taking into consideration the inhibition of cell growth by excessive light and the light intensity distributions in the cell suspensions. In these cultures, 2.1 (transformed cells) and 3.3 (host cells) kg dry cell weight per m3 were harvested at culture times of 9.0 and 10 d, respectively. These values were larger than those obtained in cultures of the respective cells at a fixed incident light intensity of 25 W/m2.     

Gene therapy for hematological malignancies

The management of avascular necrosis of the capitellum of the adolescent elbow continues to be a dilemma. This article is a critical retrospective analysis of 12 pediatric patients (mean age at surgery 14.5 years) who underwent arthroscopic debridement alone followed by early range of motion. Follow-up at a mean of 3.2 years (range, 2.0 to 5.7 years) indicated that the average flexion contracture improved from 23 degrees preoperatively to 10 degrees postoperatively. All patients had remodeling of the capitellum by plain radiographs; however, five patients had associated enlargement of the radial head. Eleven patients had minimal mechanical symptoms after the procedure and were highly satisfied. One patient had substantial enlargement of the radial head with continued loss of supination and mechanical symptoms requiring radial head resection 2 years after the index procedure. Five patients had a triangular avulsion fragment present off the lateral capsule. A statistically significant worse subjective outcome was associated with the presence of this fragment (P < .005). There were no complications.     

Human buprenorphine N-dealkylation is catalyzed by cytochrome P450 3A4

Buprenorphine (BN) is a thebaine derivative with analgesic properties. To identify and characterize the cytochrome P450 (CYP) enzyme(s) involved in BN N-dealkylation, in vitro studies using human liver microsomes and recombinant human CYP enzymes were performed. Norbuprenorphine formation from BN was measured by a simple HPLC-UV assay method, without extraction. The BN N-dealkylation activities in 10 human liver microsomal preparations were strongly correlated with microsomal CYP3A-specific metabolic reactions, i.e. triazolam 1'-hydroxylation (r = 0.954), midazolam 1'-hydroxylation (r = 0.928), and testosterone 6beta-hydroxylation (r = 0.897). Among the eight recombinant CYP enzymes studied (CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4), only CYP3A4 could catalyze BN N-dealkylation. The apparent KM value for recombinant CYP3A4 was similar to that for human liver microsomes (23.7 vs. 39.3 +/- 9.2 microM). The demonstration of BN N-dealkylation by recombinant CYP3A4 and the agreement in the affinities (apparent KM values) of human liver microsomes and recombinant CYP3A4 provide the most supportive evidence for BN N-dealkylation being catalyzed by CYP3A4.     

New electro-optical effect: optical activity of electric-field-induced twisted-nematic liquid crystal

A new electro-optical effect in nematic or nematic-cholesteric mixed liquid crystals is described which allows a 90° polarisation plane rotation of linearly polarised incident light induced by the electric field.     

Direct effect of endotoxin on the gastric mucosal microcirculation and electrical gradient

The effects of intra-arterial infusion of E. coli endotoxin at 1.0 mg. per minute on the gastric total and mucosal blood flows, electrical potential difference, and ionic fluxes across the gastric mucosa were studied in an exteriorized, chambered preparation of canine fundic stomach. Gamma-labelled microsphere technique was used in addition to venous drainage and plasma aminopyrine clearance for the measurement of total and mucosal blood flow, respectively. In spite of normal systemic blood pressure throughout the experiment, E. coli endotoxin infusion caused a significant decrease in total gastric blood flow and in the fractional distribution of flow to the mucosae. There was no significant arteriovenous shunting of microspheres. Significant reduction in potential difference and hydrogen-ion back diffusion also was noted after endotoxin infusion, possibly as a consequence of reduced mucosal blood flow. The results indicate that significant gastric mucosal ischemia can occur and may represent a mechanism in the development of gastric erosions in endotoxemia, even in the absence of systemic hypotension.