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Biomedical subjects

D P Häder

Publications and source records attributed to D P Häder.

At least 19 recordsLinked to original sources

Effects of solar radiation on the Patagonian macroalga Enteromorpha linza (L.) J. Agardh-Chlorophyceae.

The photosynthetic performance of Enteromorpha linza (L.) J. Agardh-Chlorophyceae was determined with a portable PAM instrument in situ and under seminatural radiation conditions in Patagonia, Argentina. Solar radiation was measured in parallel with a three-channel radiometer, ELDONET (Real Time Computer, Möhrendorf, Germany), in three wavelength ranges, UV-B (280-315 nm), UV-A (315-400 nm), and PAR (400-700 nm). The effective photosynthetic quantum yield decreased after 15-min exposure to solar radiation when the thalli were kept in a fixed position but recovered in the subsequent shade conditions within several hours. A 30-min exposure of free floating thalli, however, caused less photoinhibition. The photosynthetic quantum yield of E. linza was also followed over whole days under clear sky, partly cloudy and rainy conditions in a large reservoir of water (free floating thalli) and in situ (thalli growing in rock pools). Most of the observed effect was due to visible radiation; however, the UV wavelength range, and especially UV-B, caused a significant reduction of the photosynthetic quantum yield. Fluence rate response curves indicated that the species is a typical shade plant which showed non-photochemical quenching at intermediate and higher irradiances. This is a surprising result since these algae are found in the upper eulittoral where they are exposed to high irradiances. Obviously they utilize light only during periods of low irradiances (morning, evening, high tide) while they shut down the electron transport chain during intensive exposure. Fast induction and relaxation kinetics have been measured in these algae for the first time and indicated a rapid adaptation of the photosynthetic capacity to the changing light conditions as well as a fast decrease of PS II fluorescence upon exposure to solar radiation. There was a strong bleaching of chlorophyll due to exposure to solar radiation but less drastic bleaching of carotenoids.

Argentina↗

Automated biomonitoring using real time movement analysis of Euglena gracilis.

An automated biomonitoring system for early warning of pollutants in aquatic environments is described and characterized. The system uses sublethal changes in the movement behavior of the flagellate Euglena gracilis as biological endpoints. The movement is determined by real time image analysis. All parameters describing motility, velocity, orientation, and form of the cells are calculated during measurement, and changes of these parameters are interpreted as effect. By automatic dilution of the water sample, dose-effect relationships can be recorded automatically. A total measurement procedure, including control and sample measurement and filling and rinsing of the system, typically requires 8 min. Measurements with different organic and inorganic toxic compounds were performed and the calculated EC(50) values compared with literature data for the bioluminescence test with Vibrio fischeri. Also, measurements with waste water samples from different industrial plants were performed. The fast response time, the small size, the reliable image analysis system, the calculation of several endpoints, and the automatic measuring procedure are major advantages compared to other biological test systems.

Animals↗

A cyanobacterial mutant resistant against a bleaching herbicide.

The nitrogen fixing cyanobacterium Nostoc muscorum has been found to be sensitive to the herbicide SAN 6706 [4-chloro-5 (dimethylamino)-2-(a,a,a,-trifluoro-m-tolyl)-3-(2H) pyridazinon] at 30-45 microM within 15 min. The toxicity was more severe in combined nitrogen-free (Ncomb-free) medium than in a combined nitrogen medium; this enhancement was reversible by supplementation of the medium with 3 mM glucose or 5 microM ATP, serving as carbon and/or energy source in this organism. A mutant of this cyanobacterium resistant to 3 mM SAN 6706 has been isolated and characterized to perform nitrogenase activity in exogenous ATP supplemented Ncomb-free medium. However, it exhibited a moderate growth in combined nitrogen media in the absence of external ATP. The resistance factor is higher than 100. Simultaneously, this strain possesses a cross-resistance to methylamine, a well-known inhibitor of photophosphorylation, irrespective of the exogenous ATP supply. The behavior of the mutant suggests a defective phosphorylation in its photosynthetic system.

Adenosine Triphosphate↗

A cyanobacterial recombination study, involving an efficient N2-fixing non-heterocystous partner.

Many filamentous cyanobacteria fix atmospheric nitrogen under natural conditions in specialized anaerobic compartments, heterocysts, interspersed between vegetative cells, which provide protection to the O2-sensitive nitrogenase. A few unicellular cyanobacterial strains are also known to fix nitrogen aerobically at a slower rate. Filamentous cyanobacteria lacking heterocysts are not known so far to fix nitrogen. We describe the isolation and purification of a non-heterocystous filamentous cyanobacterium from the fronds of the water-fern Azolla, fixing nitrogen at 18.7+/-0.2 n moles ethylene microg Chl. a(-1) h(-1) when grown in nitrogen-free medium at a low level of oxygen between two layers of agar. This strain of Anabaena azollae has been designated as het- nif+ (non-heterocystous and nitrogen-fixing), and is found to be easily and effectively preserved in nitrogen-free medium in standard synthetic cyanobacterial nutrient medium (pH 8.5) at a continuous light intensity of 2800 lx at 25+/-1 degrees C. This het- nif+ strain is an effective donor of the nif+ marker to a het+ nif- strain of another cyanobacterium, Nostoc muscorum, when both are grown together in a recombination study.

Anabaena↗

UV effects on invertebrate and diatom assemblages of Greece.

The effects of solar radiation (PAR, UVA, UVB) on the productivity and structure of diatom and invertebrate assemblages were assessed during primary succession on artificial substrate near a rocky shore of the Saronikos Gulf, Greece. Three light treatments were performed (PAR, PAR+UVA, and PAR+UVA+UVB) at 0.5, 1.0 and 1.5 m of depth. Pennate diatoms were the major component of the developing periphytic communities during the study period. Exposure to solar UVB initially reduced the biomass and altered the structure of the diatom assemblages. The highest biomass of diatom assemblages was observed under PAR (49.2 g/m2). This value was significantly higher than the biomass of assemblages growing under PAR+UVA+UVB, but not significantly different compared to the biomass of assemblages exposed to PAR+UVA. These differences, however, did not persist at later stages. The most abundant invertebrate groups present were Polychaetes and Crustaceans. Solar UVB did not have significant effects on invertebrate biomass. Analysis of the invertebrate assemblage structure revealed time-course differences but no clear trends among the different treatments.

Animals↗

Photoprotective compounds in cyanobacteria, phytoplankton and macroalgae--a database.

A database on photoprotective compounds in cyanobacteria, phytoplankton and macroalgae has been developed. It contains information on photoprotective compounds such as mycosporine-like amino acids (MAAs), scytonemin and other not yet identified compounds reported in aquatic organisms, their habitat, the collection site and date and the reference. Further information on the absorption maxima and extinction coefficients of different photoprotective compounds as well as experimental procedures are provided (see http:/ /www.biologie.uni-erlangen.de/botanik1/index.html). The database answers the urgent need for a library on these substances and provides scientists in the field with the necessary information to identify and quantify screening pigments in different aquatic organisms from various growing sites and different collection dates.

Cyanobacteria↗

ELDONET--European Light Dosimeter Network. Structure and functions of the ELDONET server.

The European Light Dosimeter Network (ELDONET) project has been designed with the purpose of establishing an efficient system to monitor solar radiation in Europe, in as many as possible locations. This paper describes the structure of the server that collects and processes the data acquired by the different stations belonging to the network, and makes them freely available on the Internet to the scientific community. The server is able to receive data either via FTP from the Internet or via modem and to process them looking for errors or inconsistencies. Moreover, it automatically generates graphs, Web-pages and FTP archives. The server has been active for some years in testing mode and is now fully operative.

Computer Storage Devices↗

Isolation and characterisation of chemotactic mutants of Chlamydomonas reinhardtii obtained by insertional mutagenesis.

The swimming behaviour of the green flagellated protist Chlamydomonas reinhardtii is influenced by several different external stimuli including light and chemical attractants. Common components are involved in both the photo- and chemo-sensory transduction pathways, although the nature and organisation of these pathways are poorly understood. To learn more about the mechanism of chemotaxis in Chlamydomonas, we have generated nonchemotactic strains by insertional mutagenesis. The arginine-requiring strain arg7-8 was transformed with DNA carrying the wild-type ARG7 gene. Of the 8630 arginine-independenttransformants obtained, five are defective in their chemotaxis towards various sugars. Two of the mutants (CTX2 and CTX3) are blocked only in their response to xylose. Mutant CTX1 is blocked in its response to xylose, maltose and mannitol, but displays normal taxis to sucrose. Mutants CTX4 and CTX5 lack chemotactic responses to all sugars tested. CTX1, CTX4 and CTX5 represent novel chemotactic phenotypes not previously obtained using ultra-violet or chemical mutagenesis. Genetic analysis confirms that each mutation maps to a single nuclear locus that is unlinked to the mating-type locus. Further analysis of CTX4 indicates that the mutant allele is tagged by the transforming ARG7 DNA. CTX4 appears to be defective in a component specific for chemotactic signal transduction since it exhibits wild-type photobehavioural responses (phototaxis and photoshock) as well as the wild-type responses of EGTA-induced trans-flagellum inactivation and acid-induced deflagellation. Insertional mutagenesis has thus permitted the generation of novel chemotactic mutants that will be of value in the molecular dissection of the signalling machinery.

Animals↗

A novel express bioassay for detecting toxic substances in water by recording rhodopsin-mediated photoelectric responses in Chlamydomonas cell suspensions.

The influence of Cu2+, Zn2+, Cd2+, Pb2+ and formaldehyde on rhodopsin-mediated photoelectric responses in the green flagellate Chlamydomonas reinhardtii was investigated using three modifications of a recently developed population method for electrical recording (in nonoriented, phototactically preoriented (PO) and gravitactically preoriented cell suspensions). The addition of the heavy metal ions at concentrations several times lower than those known to affect swimming velocity and other physiological parameters in photosynthetic flagellates led to a rapid (one to several minutes) inhibition of the responses. Formaldehyde induced a significant temporary increase in the gravi-orientation of the cells simultaneously with an inhibition of their photoelectric cascade, photo-orientation and motility. The signals recorded in PO suspensions were more sensitive to all tested toxic substances than those recorded from nonoriented cells and indicated a switch from negative to positive phototaxis in the presence of the toxic substances. Of the two major components of the photoelectric cascade, the regenerative response was more sensitive to the tested heavy metal ions, but not to formaldehyde, than the photoreceptor current. The results obtained show that measurement of the photoinduced electrical responses in Chlamydomonas cell suspensions is a powerful novel bioassay for testing environmental pollutants in water samples.

Animals↗

Graviresponses of certain ciliates and flagellates.

Protozoa are eukaryotic cells and represent suitable model systems to study the mechanisms of gravity perception and signal transduction due to their clear gravity-induced responses (gravitaxis and gravikinesis). Among protists, parallel evolution for graviperception mechanisms have been identified: either sensing by distinct stato-organelles (e.g., the Müller vesicles of the ciliate Loxodes) or by sensing the density difference between the whole cytoplasm and the extracellular medium (as proposed for Paramecium and Euglena). These two models are supported by experiments in density-adjusted media, as the gravitaxis of Loxodes was not affected, whereas the orientation of Paramecium and Euglena was completely disturbed. Both models include the involvement of ion channels in the cell membrane. Diverse experiments gave new information on the mechanism of graviperception in unicellular systems, such as threshold values in the range of 10% of gravity, relaxation of the responses after removal of the stimulus, and no visible adaptation phenomena during exposure to hypergravity or microgravity conditions for up to 12 days.

Animals↗

Differential behaviour of two cyanobacterium species to UV radiation. Artificial UV radiation induces phycoerythrin synthesis.

Altitude is an important factor contributing to the local UV-B climate. In the European Alps solar UV-B increases approximately 21% 1000 m-1. A Nostoc muscorum (UTEX 389) originating from Scotland and a Nostoc sp. isolated from a highland lake (Yanaqocha) located 3980 m above sea level (Cusco, Perú) have been used in a study where the tolerance to UV radiation (UVR) stress of both species was determined. Following irradiation doses of 15 kJ UV (UV-A plus UV-B, equivalent to approximately 6 h exposure to unfiltered solar light at noon for a standard midlatitude region with normal ozone concentration), the viability of Nostoc sp. is 30% compared to 3% for Nostoc muscorum. UV-B induces the reduction of the number of phycobilisomes per cell, phycobilisome disassembly and/or degradation as well as phycobilisome uncoupling. Following UV exposure, phycoerythrin (PE) fluorescence emission increases dramatically in both species, indicating accumulation of PE in the phycobilisome rods. The detected increase in PE due to UVR is confirmed using a monoclonal antibody anti-PE.

Cyanobacteria↗

Electrophoretic applications of phycobiliproteins.

Phycobiliproteins are homologous chromoproteins which constitute the phycobilisomes, the light harvesting complexes of the photosynthetic apparatus in cyanobacteria, rhodophyta and cryptophyta. In the present work, phycocyanin (PC) and phycoerythrin (PE) from a Nostoc species are proposed as protein markers for electrophoretic techniques. Phycocyanin is a blue-colored phycobiliprotein; it carries phycocyanobilin as chromophoric group and is composed of two subunits, alpha and beta, with Mr of 14000 and 17000, respectively. In contrast, the PE subunits, having a similar Mr of 21000, are deep rose chromoproteins and carry phycoerythrobilin residues. Both low molecular weight phycobiliproteins are also suitable for monitoring protein blotting and the focusing time of protein samples during isoelectric focusing as internal markers. The PE subunits which form a single broad band after sodium dodecyl sulfate-polyacrylamide gel electrophoresis have different isoelectric points, and they form two visible bands when they reach their isoelectric point. The phycobilisomes constitute up to 50% of the total protein in cyanobacteria and their content in PC or PE can be up- or down-regulated by using different light conditions (chromatic adaptation).

Bacterial Proteins↗

Influence of accelerations on the spatial orientation of Loxodes and Paramecium.

The gravitactic ciliates Paramecium and Loxodes were cultivated for 15 days in space during the IML-2 spacelab mission. At dedicated times their behavioral responses to different accelerations between 10(-3) x g and 1.5 x g were investigated by using a slow rotating centrifuge microscope (NIZEMI). The threshold for gravitaxis of Paramecium was found to be at > 0.16 x g and < or = 0.3 x g. No adaptation of Paramecium to the conditions of weightlessness was observed over the duration of 15 days. Loxodes showed no graviresponses to increasing accelerations, though it demonstrated gravitaxis after return to earth.

Acceleration↗

Graviperception in the flagellate Euglena gracilis during a shuttle space flight.

During a recent space flight, gravitaxis of the unicellular photosynthetic flagellate, Euglena gracilis, was studied on board of the American shuttle Columbia. Accelerations were varied between 0 and 1.5 x g using a slow rotating centrifuge microscope (NIZEMI). The cells showed a sigmoidal response curve for the dependence of the precision of gravitaxis on acceleration which is indicative of the involvement of an active, physiological gravireceptor with a threshold at g-values < or = 0.16 x g and a saturation at g-values > or = 1 x g. No adaptation to microgravity was found during the prolonged space mission. After return the cells showed a normal gravitactic behavior at 1 x g. Since the cells are heavier than water, their swimming velocity is affected by sedimentation. The velocity distribution at different accelerations closely follows Stokes' law for sedimentation indicating that, in contrast to the ciliate Paramecium, E. gracilis, does not show any gravikinesis.

Acceleration↗

A rapid, simple method for the isolation and characterization of the photoreceptor of Dictyostelium discoideum amoebae.

A membrane-bound 45.5 kDa protein has been isolated from Dictyostelium discoideum amoebae. It shows an absorption spectrum, which closely resembles the action spectrum for amoebal phototaxis, leading to the conclusion that this protein might play an important role in the photoreception of Dictyostelium amoebae. For further characterization we employed phase partition in an aqueous polymer two-phase system, which was developed by Widell and Larsson for the separation of plasma membrane proteins of higher plants. This method clearly shows that the 45.5 kDa protein is a plasma membrane protein and not an intracellular protein. Furthermore, by using phase systems with increasing polymer concentrations, this simple and rapid purification of plasma membrane proteins allowed us to isolate the putative photoreceptor in one single step. Compared to standard biochemical methods phase partition provides an enormous facilitation of the isolation of D. discoideum membrane proteins.

Animals↗

Biochemical and spectroscopic characterization of the putative photoreceptor for phototaxis in amoebae of the cellular slime mould Dictyostelium discoideum.

Using sucrose gradient centrifugation, anion exchange chromatography on a SMART system, isoelectric focusing (IEF) and sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS PAGE), a 45.5 kDa membrane protein was isolated from amoebae of the cellular slime mould Dictyostelium discoideum. The absorption spectrum of the isolated protein corresponds well with the action spectrum for the photoaccumulation of the amoebae of Dictyostelium discoideum showing a major peak between 405 and 412 nm and some minor peaks between 500 and 600 nm. The remarkable similarity of the two spectra leads to the hypothesis that the isolated protein-pigment complex may serve as a photoreceptor for amoebal phototaxis.

Animals↗

Evidence for UV-B-induced DNA degradation in Euglena gracilis mediated by activation of metal-dependent nucleases.

It is demonstrated that in vivo irradiation with artificial UV-B for several hours significantly reduces the amount of large DNA extractable from immobilized Euglena in comparison with non-irradiated controls. This UV-B effect can be eliminated by a drastic reduction of the divalent ion concentration in the extracellular medium, i.e. the substitution of the culture medium by Tris-buffered agarose. Moreover, in vitro degradation of large DNA is demonstrated for crude protein extracts isolated from non-irradiated or UV-B-irradiated Euglena. The nuclease activity is shown for both crude protein extracts and purified nucleases; in both cases, two protein bands possessing nuclease activity are obtained with apparent molecular masses of 26 and 40 kDa and their activity is inhibited by specific nuclease inhibitors, i.e. aurintricarboxylic acid and ATP, applied at a concentration as low as 10(-8) M. Moreover, in vitro, nuclease activity clearly depends on the pH, with an optimum around pH 4.5, and on the ion composition of the extracellular medium. A strong stimulating effect is shown for Ca2+ with an optimum around 10(-4) M; this effect is potentiated by Zn2+ and Mn2+, but strongly counteracted by Mg2+ and the calmodulin inhibitors trifluoperazine and N- (6-aminohexyl)-5-chloro-1-naphthalenesulphonamide (W5). These results favour the concept which explains the lethal UV-B effect on Euglena as arising from a change in the general metabolic state of the cell and an activation of a DNA-degrading system, i.e. activation of metal-dependent nucleases (U.K. Tirlapur, D.-P. Häder and R. Scheuerlein, UV-B mediated damage in the photosynthetic flagellate, Euglena gracilis, studied by image analysis, Beitr. Biol. Pflanzen, 67 (1992) 305-317).

Animals↗