Application of high-performance liquid chromatograph-electrospray ionization mass spectrometry and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry in combination with selective enzymatic modifications in the characterization of glycosylation patterns in single-chain plasminogen activator

The application of high-performance liquid chromatography (HPLC), electrospray ionization mass spectrometry (ESI-MS) and matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and selective enzymatic deglycosylation treatments is demonstrated in the analysis of glycosylation patterns in recombinant Desmodus salivary plasminogen activator, a heterogeneous glycoprotein. The sample was initially digested with a proteolytic enzyme (endoproteinase Lys-C) and then further treated with either PNGase F to remove N-linked carbohydrates or a combination of neuraminidase and O-glycosidase to remove sialic acid and O-linked carbohydrates. By comparison of the LC-ESI-MS peptide maps for the fully glycosylated and deglycosylated samples, it was possible to unambiguously identify the sites of N-linked glycosylation as well a number of N-linked glycopeptides. The O-link glycopeptides, which are present at low level ( < 1%), were not detected prior to the deglycosylation, nor could changes in peptide elution in the map following deglycosylation be correlated with potential O-linked glycosylation sites.     

The cardiovascular effects of diazepam and of diazepam and pancuronium during fentanyl and oxygen anaesthesia

A study of the physiological behaviour of the synthetic compound Orthonil (alpha-chloro-beta-(3-chloro-o-tolyl)-propionitrile revealed a strong auxin activity in higher plants. Otherwise, Orthonil appeared not to be a herbicide-auxin. It is metabolized intensively in plant tissues. Among the identified metabolites, two compounds were detected which also exert a high auxin activity. It is concluded that at least a part of the apparent auxin activity of Orthonil may be due to alpha-(3-chloro-o-tolyl) acetic acid, one metabolite of Orthonil. Although Orthonil strongly stimulates elongation growth, this growth is not accompanied by a stimulated ethylene evolution as is the case with other auxins. A possible metabolic pathway of Orthonil is discussed.     

The citric acid fermentation by Aspergillus niger: regulation by zinc of growth and acidogenesis

The citric acid fermentation by Aspergillus niger is divided into two consecutive phases, a growth phase when the cells proliferate but do not accumulate citrate, followed by an accumulating phase when they produce citrate but do not proliferate, or else do so at a much reduced rate. When studied in a low sucrose (0.4-0.8%) mpinimal salts medium the growth-accumulation alternative was controlled by the concentration of zinc: high zinc (about 1-2 muM) maintained growth phase, while at low zinc (below 1 muM) growth became limited by zinc deficiency and the cultures passed into accumulating phase. Addition of zinc to accumulating cultures resulted in their reversion to growth phase. Iron, manganese, calcium at concentrations as high as 5-10 muM had no influence on either growth or citrate accumulation. These results suggest that zinc plays a role in the regulation of growth and citric acid accumulation.     

Discovery of insect anti-juvenile hormones in plants.?2U

Two simple chromenes with anti-JH activity have been isolated and identified from the bedding plant Ageratum houstoianum. By contact and fumigation these compounds induce precocious metamorphosis and sterilization in several hemipteran species of insects. Certain holometabolous species are sterilized, forced into diapause, or both. Each of these biological actions is equivalent to removal of the corpora allata, which produce the JH's, and is reversible by treatment with exogenous JH. Thus, the action of these compounds is to stop the production or depress the titer of the JH's. To our knowledge, this is the first discovery of anti-JH, and we hope it will guide the way to the emergence of a fourth generation of safe and insect-specific pesticides.     

Two-beam Coupling Modules for Photorefractive Optical Circuits

Modules that perform photorefractive two-beam coupling operations have been built, characterized, and tested. These portable modules, interconnected by fiber optics, dispense with the need for repeated alignment and greatly facilitate the prototyping of complex signal- or image-processing photorefractive circuits. To evaluate the performance of the modules in a photorefractive circuit, we interconnected them in the feature extractor configuration: a ring configuration composed of two modules that selects the strongest signal within the signals presented on its input. With two signals at the input, an output contrast ratio of 45.4 dB is obtained for an input contrast ratio of 5 dB.     

Restoration of squamous mucosa after ablation of Barrett's esophageal epithelium

BACKGROUND: Antireflux therapy has generally failed to induce regression of Barrett's epithelium. It was hypothesized that squamous epithelium could be restored if the columnar tissue was ablated while gastric acid secretion was suppressed. METHODS: Ten white men with Barrett's esophagus received 40 mg of omeprazole daily. Thereafter, every 2-5 weeks they underwent videotaped endoscopies to argon laser photoablate columnar tissue, obtain biopsy specimens, and assess results. Squamous re-epithelialization was assessed by correlation of videotapes and directed biopsies. RESULTS: Patients had one to eight areas ablated, totaling 0.5-12.0 cm2. Videotape assessments were corroborated by biopsy in all but one instance. Thirty-eight of 40 treatment locations partially or completely re-epithelialized with squamous tissue. Squamous regrowth appeared to occur by spread from contiguous squamous borders and de novo from glandular tissue. Regrowth was influenced by the extent of squamous borders and completeness of ablations. Nonablated glandular tissue persisted beneath squamous epithelium. CONCLUSIONS: Ablation of Barrett's epithelium and suppression of acid secretion facilitated squamous re-epithelialization. A progenitor cell within the metaplastic tissue has the potential to differentiate normally.     

trans-2-phenylcyclopropylamine is a substrate for and inactivator of horseradish peroxidase

Horseradish peroxidase (HRP) is well known for mediating the electron-transfer oxidation of electron-rich aromatic 'donors' such as phenols and anilines, but has not been described to oxidize aliphatic amines. We here confirm the inability of HRP to oxidize typical aliphatic amines, even those which would exist significantly as free bases at the operative pH. In contrast, trans-2-phenylcyclopropylamine (2-PCPA) is both a substrate (turnover product is cinnamaldehyde) and a time-dependent inactivator of HRP. These activities of 2-PCPA are consistent with either a concerted or rapid sequential one-electron-oxidation/ring-opening to give an intermediate capable of covalent binding to the enzyme. 2-PCPA is the first known example of a simple aliphatic amine which serves as a substrate for HRP under turnover conditions.