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The present paper describes a novel generation of microchips suitable for fluorescence-based assays, such as cDNA, oligonucleotide, or protein microarrays. The new transducers consist of a fully corrugated surface coated with a thin layer of [Ta.sub.2][O.sub.5] as a high refractive index material. Tuning of the incident excitation light beam to abnormal reflection geometry results in a confinement of the energy within the thin metal oxide layer. Consequently, strong evanescent fields are generated at the surface of these microchips and fluorophores located within the fields showed up to a 2 order of magnitude increase in fluorescence intensities relative to the epifluorescence signals. We have attributed this phenomenon as evanescent resonance (ER). Due to the surface architecture, propagation distances of the incident energy and fluorescence photons are in the micrometer range, thus preventing cross talk between adjacent regions. ER microchips offer a significant increase in fluorescence intensities in both 'snapshot' fluorescence setups and commercial fluorescence scanners. The underlying principle of the novel chips is explained, and quantitative data on the fluorescence enhancement are provided. To demonstrate their potential, the novel chips are used to investigate the dependence of expression levels from metabolic genes in rat liver on drug treatment. In contrast to competitive hybridization, labeled samples were hybridized to individual ER microchips, and changes were observed by comparing with normalized data from different chips. Results obtained in gene expression profiling experiments with phenobarbital-treated rats are shown.

Solution-state and solid-state NMR spectroscopic analyses confirm the formation of a covalent intermediate during catalysis by the enzyme uridine diphosphate-N-acetylglucosamine (UDP-NAG) enolpyruvyl transferase from Enterobacter cloacae. This enzyme promotes the transfer of an enolpyruvyl group from phosphoenolpyruvate to the 3-hydroxyl of UDP-NAG, forming enolpyruvyl UDP-NAG and phosphate. The covalent O-phosphothioketal enzyme intermediate has a methyl group at C-3, with the C-2 being quaternary.

The general control transcriptional regulator gene cpcA of Aspergillus niger was cloned by complementation of a Saccharomyces cerevisiae Δgcn4 mutant strain. The encoded protein conferred resistance to amino acid analogues when expressed in yeast. Disruption of cpcA in A. niger resulted in a strain which is sensitive towards 3-aminotriazole and fails to respond to amino acid starvation, cpcA encodes a transcript of ≈2400 nucleotides in length that includes a 5′ leader region of 900 nucleotides. The 5′ leader region contains two small open reading frames, suggesting translational control of gene expression. Steady-state mRNA levels of cpcA increase by a factor of three upon amino acid starvation. The coding region of cpcA is interrupted by a 57 bp intron and the deduced amino acid sequence displays an ≈30% overall identity to yeast GCN4p and Neurospora crassa cpc1p. Critical amino acid residues of the transcriptional activation domains of GCN4p are conserved in cpcAp. The basic DNA-binding domain shows up to 70% amino acid sequence identity to other basic zipper (bZIP)-type transcriptional activators, cpcAp binds specifically to a GCN4p recognition element in gel retardation experiments. The C-terminal dimerization domain encodes a leucine zipper with only a single leucine residue. [ABSTRACT FROM AUTHOR]

Gastrointestinal ( GI) pH and temperature profiles under fasted-state conditions were investigated in two studies with each 10 healthy human subjects using the IntelliCap® system. This telemetric drug delivery device enabled the determination of gastric emptying time, small bowel transit time, and colon arrival time by significant pH and temperature changes. The study results revealed high variability of GI pH and transit times. The gastric transit of IntelliCap® was characterized by high fluctuations of the pH with mean values ranging from pH 1.7 to pH 4.7. Gastric emptying was observed after 7-202 min (median: 30 min). During small bowel transit, which had a duration of 67-532 min (median: 247 min), pH values increased slightly from pH 5.9-6.3 in proximal parts to pH 7.4-7.8 in distal parts. Colonic pH conditions were characterized by values fluctuating mainly between pH 5 and pH 8. The pH profiles and transit times described in this work are highly relevant for the comprehension of drug delivery of solid oral dosage forms comprising ionizable drugs and excipients with pH-dependent solubility. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:2855-2863, 2015 [ABSTRACT FROM AUTHOR]