In situ analysis of C. elegans vitellogenin fusion gene expression in integrated transgenic strains: effect of promoter mutations on RNA localization

Expression of the Caenorhabditis elegans vitellogenin (vit) genes is initiated at the larva-to-adult molt in all of the 30 to 34 nuclei of the hermaphrodite intestine. A series of strains in which DNA carrying a vit fusion gene was integrated at low copy number was analyzed by in situ hybridization to determine whether the transgene showed the same tissue-specific expression. Strains with only 247 bp of 5'-flanking DNA accumulated the mRNA product of the introduced vitellogenin gene only in the adult hermaphrodite intestine, and uniformly in all of the intestinal cells. When similar strains carrying vit fusion genes with promoter modifications were tested, no loss of tissue specificity was observed. Surprisingly, however, strains with modified promoters that resulted in reduced levels of expression displayed a novel pattern of transgene RNA localization within their intestines. Strains with severe promoter defects accumulated the transgene mRNA in the central part of the intestine but lacked the mRNA at both ends. Those with less severe promoter mutations lacked the transgene mRNA only in the most anterior intestinal cells. We hypothesize that genes with altered promoters require higher activator concentrations to express the reporter gene, thus revealing an inherent asymmetry in activator levels, lowest in the anterior cells and highest in the central cells of the intestine.     

Phosphorylation of contractile proteins in heart and skeletal muscle

Contractile performance of cardiac and skeletal muscles may be regulated by cyclic AMP or Ca2+, two second messengers that stimulate the phosphorylation of specific myofibrillar proteins. Cyclic AMP-dependent protein kinase catalyzed the rapid phosphorylation of a single site in the inhibitory subunit of cardiac troponin in vitro and in perfused hearts. Skeletal muscle troponin was not phosphorylated by this enzyme in vivo. Although there was a correlation between cardiac troponin phosphorylation and the positive inotropic response to catecholamines, a biochemical mechanism that could account for a functional relationship between the two processes has not been discovered. Phosphorylation of skeletal muscle myosin was catalyzed by myosin light chain kinase in the presence of Ca2+ and the ubiguitous, multifunctional Ca2+-dependent regulator protein (CDR). The activation of kinase activity appeared to proceed via a trimolecular reaction process in which Ca2+ bound to CDR and the Ca2+.CDR complex then interacted with the enzyme. In rat extensor digitorum longus muscle, a 1 sec tetanic contraction resulted in phosphorylation of myosin light chain with the maximal phosphate incorporated 20 sec after the contraction. The light chain phosphate content declined slowly and correlated to post-tetanic potentiation of isometric twitch tension. Phosphorylation of skeletal muscle myosin may be important in modulating contraction.     

4-Methylimidazol in Caramel und caramelgef?rbten Lebensmitteln

Zusammenfassung Es wird eine spezifische und kochempfindliche Methode zur Bestimmung von 4-Methylimidazol (4-MeI) in Caramel und caramelgefärbten Lebensmitteln beschrieben. Nach der Extraktion wird 4-MeI gaschromatographisch mit einem stickstoffspezifischen elektrochemischen Detektor (Hall) bestimmt. Als weniger spezifische Alternative kann ein thermoionischer Detektor eingesetzt werden. Bei Zusätzen von 5–200 mg 4-MeI/kg Karamel beträgt die Wiederfindungsrate 90–94%.Die Nachweisgrenze der Methode liegt unterhalb 0,1 mg 4-MeI/kg Caramel; in karamelgefärbten Lebensmitteln können weniger als 0,01 mg/kg nachgewiesen werden.

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4-methylimidazole in caramel and caramel-colored foodsDetermination by gas-liquid-chromatography with nitrogen-specific detectors

Summary A specific and highly sensitive method for the determination of 4-methylimidazole (4-MeI) in caramel and caramel-colored foods is described. After extraction the amount of 4-MeI is determined by gas-liquid chromatography with a nitrogen-specific electrochemical detector (Hall). As a less specific alternative a thermoionic detector can be used. With additions in the range of 5–200 mg 4-MeI/kg caramel, recoveries of 90–94% are obtained.The limit of detection is <0.1 mg 4-Mel/kg caramel; in caramel-colored foods less than 0.01 mg/kg can be detected.

     

Isolation and characterization of the rodlet layer of Trichophyton mentagrophytes microconidial wall

The rodlet layer of the microconidial wall of Trichophyton mentagrophytes was isolated and partially characterized. The purified microconidial walls were first extracted with urea (8M), mercaptoethanol (1%), and sodium dodecyl sulfate (1%) followed by enzymatic digestion with glusulase (snail intestinal enzymes) and purified (1 leads to 3)-beta-D-glucanase and chitinase. The purified rodlet layer was 15 to 30 nm thick and accounted for approximately 10% of the original wall weight. The pattern of rodlet patches, as revealed by electron microscopy of freeze-etched preparations of the isolated layer, was essentially the same as that observed on the intact microconidial wall. The rodlet layer was found to be resistant to most of the common organic solvents, cell wall lytic enzymes, mild acid treatments, and surface-active agents, but was solubilized in boiling 1 N NaOH with concomitant disorientation of the rodlet patterns. A melanin or melanin-like pigment appeared to be intimately associated with this rodlet layer and was solubilized during a hot-alkali treatment. Protein (80 to 85%) and glucomannan (7 to 10%) were the major components of the rodlet layer. The rodlet layer did not contain any appreciable amounts of lipid or phosphorus.     

Thermodynamic Properties of Nonstoichiometric Yttria-Stabilized Zirconia at Low Oxygen Pressures

Thermodynamic properties of 8-mol%-yttria-stabilized zirconia have been determined in the 810° to 1040°C temperature range at low Po₂. A high-temperature solid-state coulometric titration method was used. The mass action constant, K_ma, can be represented at low Po₂ as K_ma=0.677 exp [(–3.98 ±0.03 eV)/kT].     

Analysis of Digital System Performance in EAM-Based Photocurrent Driven Wavelength Converter

This work describes the factors that influence the digital system performance of a monolithically integrated photocurrent driven wavelength converter. For an optimized input power to the receiver section of the device, experiments show <1-dB power penalty for conversion between 1548 and 1563 nm at 10 Gb/s. Under optimized conditions, performance is limited by the output extinction ratio of the converted signals