The 1-Hz fluorometer: A new approach to fast and sensitive long-term studies of active chlorophyll and environmental influences

A new instrument for environmental monitoring, called at 1-Hz fluorometer, provides two modes of application. First, it enables a quantitative determination of algal concentrations down to 20 ng/l. Second, it can be used as a biosensor for changes in environmental conditions. The distinction between the signals from living chlorophyll-containing algae and other fluorescent material is achieved by using two modulated light-sources resulting in a mean fluence rate of 200 μE. The measuring light induces changes in chlorophyll fluorescence (yield) with a frequency of 1 kHz, and the actinic light modulates the redox state of the quenchers of PS II with a frequency of 1 Hz. This leads to a modulation of the yield which is detected by two phase-sensitive rectifiers (double correlation). Measurements from different sites in a river, and in the Baltic and North Seas, show that correction by the built-in simultaneously-measured attenuation is necessary in order to obtain values which are identical with those determined by a photometric analysis (Uvikon 860). This applies if the transmission becomes less than about 95%. Suspensions ofDunaliella salina exposed to ammonia and phosphate were used for illustrating the usage for environmental monitoring. It is shown that this system can measure changes in the chlorophyll fluorescence of living algae caused by changes in concentration of ammonia down to 1 μg/l and of phosphate down to 10 μg/l.     

Genetic variation for foot-rot and Fusarium head-blight resistances among full-sib families of a self-incompatible winter rye (Secale cereale L.) population

The amount of genetic variation for resistance to foot rot caused by Pseudocercosporella herpotrichoides, Fusarium spp., and Microdochium nivale and for resistance to head blight caused by Fusarium culmorum are important parameters when estimating selection gain from recurrent selection in winter rye. One-hundred and eighty-six full-sib families of the selfincompatible population variety Halo, representing the Petkus gene pool, were tested for foot-rot resistance at five German location-year combinations (environments) and for head-blight resistance in three environments with artificial inoculation in all but one environment. Foot-rot rating was based on 25 stems per plot scored individually on a 1–9 scale. Head-blight resistance was plotwise scored on a 1–9 scale and, additionally, grain-weight per spike was measured relative to the non-inoculated control plots. Significant estimates of genotypic variance and medium-sized heritabilities (h ²=0.51–0.69) were observed in the combined analyses for all resistance traits. In four out of five environments, the amount of genetic variance was substantially smaller for foot-rot than for head-blight rating. Considerable environmental effects and significant genotype-environment interactions were found for both foot-rot and head-blight resistance. Coefficients of error-corrected correlation among environments were considerably closer than phenotypic correlations. No significant association was found between the resistances to both diseases (r=-0.20 to 0.17). In conclusion, intra-population improvement by recurrent selection should lead to substantial higher foot-rot and head-blight resistances due to significant quantitative genetic variation within Halo. Selection should be carried out in several environments. Lack of correlation between foot-rot and head-blight resistance requires separate infection tests for improving both resistances.     

Intersegmental Coordination: Lessons from Modeling Systems of Coupled Non-Linear Oscillators

SYNOPSIS. Intersegmental coordination of both vertebrates andinvertebrates is poorly understood primarily because so littleis known about the substrate of the neural underpinnings andhow the elements interact to produce the complex timing relationshipsrequired by the organism. We describe here a systems approachcombining experimental and theoretical treatment of the lampreylocomotor central pattern generator. The central pattern generatoris viewed as a chain of coupled non-linear oscillators correspondingto segmental burst generators. We analyze various coupling schemesin terms of their ability to produce a stable traveling wavesimilar to that seen in the isolated spinal cord and the intactanimal. The role of long coordinating neurons is particularlydiscussed.     

Stereospecific high-performance liquid chromatographic assay of ibuprofen: improved sensitivity and sample processing efficiency

A high-performance liquid chromatographic (HPLC) assay suitable for the analysis of the enantiomers of the non-steroidal anti-inflammatory drug ibuprofen (IB) in plasma was developed. Following the addition of racemic fenoprofen as internal standard (I.S.), samples are acidified and extracted with a mixture of isooctane—isopropanol (95:5, v/v). After evaporation of the organic layer, the drug and I.S. are derivatized with S-(−)-1(1-naphthyl)ethylamine (S-NEA) after addition of ethyl chloroformate as the coupling reagent. Ethanolamine is added 3 min after the addition of S-NEA to react with the excessive ethyl chloroformate. The resultant diastereomers corresponding to IB and I.S. were chromatographed at ambient temperature on a 100 mm × 4.6 mm I.D. C₁₈ reversed-phase column using acetonitrile—water—acetic acid—triethylamine (60:40:0.1:0.02) as the mobile phase pumped at a flow-rate of 1.2 ml/min. Detection of the fluorescent chromophore was at 280 and 320 nm for excitation and emission, respectively. The suitability of the assay for clinical pharmacokinetic studies of IB was determined by the analysis of plasma samples obtained from a healthy volunteer, following administration of a single 400-mg oral dose of racemic IB.     

Properties of aspartate racemase, a pyridoxal 5'-phosphate-independent amino acid racemase.

Aspartate racemase from Streptococcus thermophilus contains no pyridoxal 5'-phosphate or other cofactors such as FAD, NAD+, and metal ions. It was affected by neither carbonyl reagents such as hydroxylamine nor sodium borohydride but was strongly inhibited by iodoacetamide and other thiol reagents. Aspartate, cysteate, and cysteine sulfinate were the only substrates. The Km values for L- and D-aspartate were 35 and 8.7 mM, respectively. The enzyme catalyzed the exchange of alpha-hydrogen of the substrate with the solvent hydrogen. Racemization of L-aspartate in 2H2O showed an overshooting in the optical rotation of aspartate before the substrate was fully racemized. This shows that the removal of alpha-hydrogen of the substrate is at least partially rate-determining. When L- or D-aspartate was incubated with aspartate racemase in tritiated water, tritium was incorporated preferentially into the product enantiomer. The results strongly suggest that aspartate racemase contains two hydrogen acceptors.     

A novel 145 kd brain cytosolic protein reconstitutes Ca(2+)-regulated secretion in permeable neuroendocrine cells.

The regulated secretory pathway is activated by elevated cytoplasmic Ca2+; however, the components mediating Ca2+ regulation have not been identified. In semi-intact neuroendocrine cells, Ca(2+)-activated secretion is ATP- and cytosol protein-dependent. We have identified a novel brain protein, p145, as a cytosolic factor that reconstitutes Ca(2+)-activated secretion in two neuroendocrine cell types. The protein is a dimer of 145 kd subunits, exhibits Ca(2+)-dependent interaction with a hydrophobic matrix, and binds phospholipid vesicles, suggesting a membrane-associated function. A p145-specific antibody inhibits the reconstitution of Ca(2+)-activated secretion by cytosol, indicating an essential role for p145. The restricted expression of p145 in tissues exhibiting a regulated secretory pathway suggests a key role for this protein in the transduction of Ca2+ signals into vectorial membrane fusion events.     

Kinetics of Nitrosomonas europaea at extreme substrate,product and salt concentrations

Summary Nitrification of ammonia in concentrated waste streams is gaining a lot of attention nowadays. Nitrosomonas europaea is the predominant ammonia-oxidizing species in these environments. Prediction of the behaviour of a pure culture of N. europaea (ATCC 19718) under conditions prevailing in concentrated waste streams was the aim of this study. The initial oxygen consumption rate of a concentrated cell suspension was used as a rapid assay to measure the effects on N. europaea under various conditions. Several relationships, based on Michaelis-Menten kinetics, were derived. They describe the behaviour of N. europaea at substrate (NH₄ ⁺), product (NO₂ ^– and K⁺, Na⁺, SO _inf4 ^sup2– , NO₃ ^–, Cl^–) concentrations up to 500 mol/m³ and pHs ranging from 6.5 to 8.5. High concentrations of ions inhibited N. europaea but specific substrate inhibition was not observed. Product inhibition was strongly pH-dependent and severe inhibition was found at pH 6.5. Correspondence to: J. H. Hunik