Volume 33, Issue 1-2

In several series of 212 samples, higher fungi, especially from the genus Amanita and related genera, were analyzed by atomic absorption spectroscopy on their content of the trace metals vanadium and molybdenum. Only vanadium was found to be present in high concentration in the fly agaric mushroom (Amanita muscaria), while 22 of its related species showed the normal V-content of the higher fungi tested. A correlation with the presence of molybdenum could not be detected. A special enrichment of Mo was not found in higher fungi. The analysis of several fruit bodies from A. muscaria showed that the highest V-content was present in the bulb and in the lamellae, both contents increasing with age, while the spores contained only 1 - 2% of the lamellar vanadium.

Zinc, copper and cadmium concentrations in various organs of 30 roadies (Rutilus rutilus L.) from the Neckar and Elsenz Rivers were investigated. The contents of all three metals are elevated in all fishes; however, only Cd in older fishes was found to exceed the tolerance limit for human health in a section of the Neckar River, which until 1973 was heavily contaminated with Cd from industrial wastes. The higher degree of Zn-pollution in the Elsenz River is also clearly expressed in the Zncontents of the Elsenz fishes. The following order of sequence of metal concentrations (independant of the collection area and the degree of contamination) could be observed: Zn and C d: muscle < liver <C kidney Cu : muscle < kidney < liver Our findings demonstrate the importance of fishes as indicator organism for heavy metal contamination. They also clearly implicate that for ecological rather than for human toxicological investigations the organ in which a specific metal tends to preferentially accumulate (p. ex. Zn and Cu in the kidney; Cu in the liver; Pb in the skeleton; Hg in lipids) is the best indicator of that metal. As a consequence specific fish organs should be increasingly used to determine the extent of heavy metal contamination in aquatic systems.

Weak microwave irradiation of aqueous yeast cultures was found to affect their growth rate in a frequency-selective manner. Depending on frequency (near 42 GHz), both increases and de­ creases of the growth rate were observed. The resonance bandwidths are of the order of 0.01 GHz. Simple thermal effects can be excluded. These findings support theoretical predictions of coherent molecular oscillations activating metabolic processes.

We have used the CNDO/2-CI method to calculate the isopotential maps for the fundamental state and the direction of the polarizations corresponding to several singlet transitions in the mole­ cules of guanine, cytosine and tautomeric forms. We have also calculated the times for proton tunneling and equilibrium constants between both conformers corresponding to the fundamental and some excited states. The results have been compared with the available experimental data.

Fat-praeparates of cow and pork have been investigated by small-angle X-ray scattering methods. Dependent on the temperature these fats show distinct X-ray interference patterns in the small-angle region. Hence complete phasediagrams of the fats in the temperature region between -20 °C and 55 °C could be estimated. Using the intensity distribution of the X-ray scattering the electron-density distribution along one direction of the ground-cells of the fat crystals were calculated.

Diffraction studies on natural 1,2-dipalmitin and on analogs, including those based on the configurational isomers of cyclopentane-1,2,3-triol reveal that the 1,2-diglycerides crystallize from solvent with chain methylene packing identical to the monoclinic form of even-chain alkanes. The chains probably are folded back in “hairpin” fashion as found in phospholipid crystal structures. Acyl shifts are observed to occur in the crystalline solid state at room temperature to give the 1,3-diglyceride. Analogs based on the above-mentioned cyclitols show that isomers with adjacent chains trans to the ring (possibly extended chain packing) or with chains cis to the ring (“hair­ pin”) crystallize readily. Both possibly extended chain configurational isomers have the α-form as well as β′-forms and a β-polymorph. The hairpin isomers each give a β′-polymorph but only the all-cts isomer gives an α-form.

The utility of analogs to glycerol-containing lipids based on the configurational isomers of cyclo-pentane-1,2,3-triol for ab initio crystal structure analysis via electron diffraction is assessed further. Such analogs of tripalmitin are examined with the natural triglyceride via low angle X-ray diffraction. The 1,3/2 (all-trans) and 1,2/3 (cis-trans) analogs give long spacing dimensions some 23 Å greater than found for the β-2 form of the natural compound, consistent with the long spacing observed for a β-3 form. The 1,2,3/0 (all-cis) analog gives a long spacing near that of the a-form of the triglyceride. Reflection electron diffraction measurements on the 1,3/2 and 1,2/3 analogs reveal a chain tilt near 60° for both and untilted chains for 1,2,3/0. A more accurate tilt determination from X-ray long spacings of the homologous series of 1,3/2 pseudotrilaurin to 1,3/2 pseudo-tripalmitin confirm the 67° tilt expected for the β-3 form. Therefore, given the same T,, methylene subcell, the molecular packing is very close to natural triglycerides. The subtle influences of the cyclopentane ring are overcome for 1,3/2 analogs based on stearic and arachidic acids. This emphasizes the utility of these structural analogs for ab initio crystal structure determinations of glycerol containing lipids.

Condensation of cytidine or uridine with ethyl levulinate leads to the acetals 1a/2a. The reac­tion would be expected to give mixtures of diastereoisom ers. As shown by 1 H and 13 C NMR spectroscopy only one diastereoisomer is formed. By spectroscopic comparison of 1a/2a with the corresponding adenosine acetal the absolute configuration of the new chiral centre was found to be R. The acetal m ethyl group of 1a/2a in exo-location can serve to distinguish the two m ethyl signals of O-2′,3′-isopropylidenecytidine and -uridine in the NM R spectra. On alkaline hydrolysis of the esters the acids 1b and 2b are formed, which can be condensed through their carboxyl groups with 6-aminohexylagarose. The affinity resins 3 and 4 contain 7.1 μmol and 7.6 μmol ligand/g moist gel respectively. A biospecificity of the new polymers to cytidine-and uridine converting enzymes is expected.

The membrane-bound, solubilized, and trypsin-treated forms of Mg, Ca-ATPase from E. coli are inhibited by ruthenium red[R R ]. The inhibition is noncompetitive and is reduced at higher sub strate concentrations. n-Butanol-extracted ATPase is not inhibited by ruthenium red and is not activated by KCl.

1. The time course of activity of soluble and microsomal phenylalanine ammonia-lyase (PAL) was studied in dark grown cell cultures of soybean (Glycine max). A distinct activity increase of PAL in the soluble and microsomal fraction occurred prior to the stationary phase of the cell culture. Cinnamic acid p-hydroxylase and NADH : cytochrome c reductase, too, exhibited maximal activity in the log phase, 5 days after the transfer of soybean cells to fresh culture medium. 2. Upon subfractionation of the once washed microsomal fraction by sedimentation velocity centrifugation on a sucrose gradient, membranes of the endoplasmic reticulum could be separated from fractions containing mainly membranes from the Golgi apparatus or plasma membranes, respectively. PAL and cinnamic acid p-hydroxylase were found in fractions of endoplasmic reticulum whereas no activity of either enzymes could be detected in fractions containing Golgi apparatus or plasma membranes. 3. Repeated washing of microsomal fractions led to a residual membrane-bound PAL representing about 1% of the total PAL activity of the cells. This residual membrane-bound activity could be solubilized almost completely by Triton X-100 or digitonin at concentrations of 0.5 - 5%.

1. Liver and serum aspartate aminotransferase (GOT) and alanine aminotransferase (GPT) activities were measured in a hibernating desert lizard, Uromastix hardwickii. The levels of both enzymes were found to be lower in hibernation than during the active period, particularly in the liver. 2. After intramuscular injection of 2 mg of cortisone acetate there was a rapid rise in the levels of these enzymes with a peak of 18 hours (GOT) and 12 hours (G P T). 3. The response of both enzymes to cortisone was much greater during the active period than during hibernation. 4. GOT showed a much more rapid and greater response to cortisone than GPT. This is in contrast to the response of rat liver where GPT is more responsive to this hormone. 5. These studies indicate that the transferase enzymes of this lizard differ from those of the rat in their sensitivity and time of response to cortisone.

Anomeric configuration of oligosaccharides usually is established by specific glycosidases. For this purpose detergents achieving water solubility of primarily insoluble glycosphingolipids as substrates have been replaced by delipidated hum an serum high density lipoproteins. The new method, tested by several well characterized glycosphingolipids and glycosidases, finally was applied to the evaluation of anomeric structures of two blood-group B active glycosphingolipids [ceramide hexa-saccharide (B-I) and ceramide octasaccharide (B -II)] from hum an erythrocyte membranes. In both B-I and B-II, α-glycosidic linkage was dem onstrated for the term inal galactose and fucose residues. β-glycosidic linkage has been evaluated for backbone saccharides. Together with the results pre­ viously obtained by composition analysis, linkage analysis and sequence analysis the following complete structure can be established: B -I: Galα1 → 3Gal (2 ← 1αFuc)β1 → 4GlcNAcβ1 → 3Galβ1 → 4Glcβ1 → 1Cer; B-II: Galα1 → 3Gal (2 ← 1αFuc)β1 → 4GlcNAcβ1 → 3Galβ1 → 4GlcNAcβ1 → 3Galβ1 → 4Glcβ1 → 1Cer.

The num ber of SH-groups of the soluble proteins (Prot-SH) of 12 different types of animal tum ors and of cultivated chicken fibroblasts increases with decreasing doubling tim e (td). Between Prot-SH and td an inverse association was found with a significance level of 99.5%. After 30 min incubation in vitro, 4-hydroxypentenal (HPE) reacts with different am ounts of the Prot-SH of the investigated cells and tissues. It was found that the absolute, as well as the per­ cental amounts of HPE-reactive Prot-SH increase with decreasing td, each of the respective inverse associations was found to be highly significant. The tum ors were collected in three groups, each with a corresponding range of td and a mean value for Prot-SH . Taking the Prot-SH of the slowest growing tissue (DENA-hepatoma) as the reference value, the increments of Prot-SH were cal­ culated for each of the three groups and were found to be strikingly similar to the number of HPE-reactive Prot-SH. The single Prot-SH increm ents of the diverse cell and tissue types compared with DENA-hepatoma were found to be highly significantly and inversely associated with the respective Prot-SH decreases caused by HPE. Hence it may be concluded that HPE reacts preferably with those of the Prot-SH which are of some functional importance for tum or growth and, moreover that the attack on those thiols is the more effective the faster the tumor is growing.

Various strains (wild-type and mutants) of Schizosaccharomyces pombe were subjected to sporulation conditions, and analysed with regard to changes in their protein patterns. Trisbuffer-soluble proteins (TS) and DNA-binding proteins (DB) were denatured and separated by polyacrylam ide electrophoresis. A fter endogenous nitrogen depletion (the apparent trigger of conjuga­tion and sporulation) the following alterations were observed: larger TS-proteins were proteolytical-ly degraded; the pattern of DB-proteins underwent several selective changes (reduction, enforce­ ment, or new appearance of individual bands) ; an endonucleolytic DNase activity appeared. Most of these changes were expressed before conjugation, and did not appear in sterile strains. They could, however, be provoked by an abrupt shift to nitrogen-free medium, even in sterile strains. Inhibition studies with cycloheximide revealed at least two phases of protein synthesis specifically needed for sporulation: a very sensitive phase before conjugation, and a more resistant phase lasting until 0.5 h before spore form ation. Medium exchange between heterothallic cultures (h + and h - ) effected certain sporulation-specific alterations of the DB-protein pattern, indicative of diffusible mating-type factors in the culture fluid.

Psoralen photosensitization of L 1210 cells has a strong effect on DNA and RNA syntheses and this result appears connected with the psoralen photobinding to DNA. Protein synthesis is less sensitive and its inhibition seems due to a different photochemical interaction, very likely to the psoralen photobinding to RNA. A combined therapy using cyclophosphamide and L 1210 cells psoralen-photoinactivated was perform ed after the leukaem ia transplant, showing a significant decrease in mortality, even in comparison with the simple treatment with the alkilating drug.

1) DNA-protein complexes are supposed to be original constituents of the membrane of Ehrlich ascites tum or cells. These complexes can be attacked at the surface of viable cells by DNase or protease. The DNA is partially embedded in protein structures. 2) The net charge of this complex is of major importance for the RNA uptake capacity of the cells. Negatively charged DNA which is situated at the surface hinders RNA uptake. This is the explanation for the stimulation of RNA uptake by DNase or the decrease in RNA uptake after protease treatment. 3) Upon treatment of DNA-dcficient complexes with homologous or heterologous DNA the original RNA uptake capacity of the cells is restored but the original conformation of the complex cannot be regained. 4) The DNase action on the complex is tem perature dependent in a sigmoidal fashion. It is m arkedly slowed down at tem peratures below 12 °C. This implies that structural dianges in the complex occur at this transition tem perature which make surface DNA susceptible to DNase. This effect can only be observed in original structures but not in reconstituted ones. 5) Polyanion treatment of the cells [poly (L-lysine) ] which increases their RNA uptake capacity, most probably does not interact with the DNA-protein complex. Poly (L-lysine) appears to act at other m em brane sites. 6) The DNA-protein complex has been investigated entirely in situ , i. e. situated in the membrane of viable cells.

Protons are released from unilluminated chloroplasts after the addition of Mg 2+ . The greatest number of protons was measured at pH 6.5 and corresponded to 0.6 mol protons per mol chlorophyll.

9-Aminoacridine (9AA) binds to photosynthetic m em branes of unilluminated chloroplasts in low-salt media. The binding was insensitive to the uncouplers of photophosphorylation. The apparent binding constant was 140 μM . The binding isotherm as a function of 9AA concentration was sigmoid, and approximately 3 mol 9AA/mol chlorophyll was bound at saturating concentrations of 9AA. Addition of Mg 2+ partially reversed the binding of 9AA in chloroplasts in the dark as observed by a Mg 2+ -induced increase of 9AA fluorescence as well as by spectrophotom etric measurements of free 9AA. It appeared, however, that use of fluorescence techniques for m easuring free 9AA introduced an error in the estimation of the m agnitude of binding, particularly at low concentration of 9AA ( < 7 5 μM). This is probably due to change in fluorescence yield of membrane-bound 9AA on addition of cations. The nature of the binding of 9AA to the thylakoid membranes and the effects of Mg 2+ thereupon suggest that both chemical binding of cations and screening of surface charge of the mem branes should be considered in discussing the mechanism of cation action on chloroplast structure and function. Interpretation of these data with respect to heterogeneity of sites of cation action upon or within chloroplast membranes is discussed.

Studies of the rate and efficiency of H 2 production by a reconstituted system composed of washed chloroplasts, ferredoxin, and hydrogenase reveal th at a significant proportion of the low potential electrons are leaked to H 2 O 2 production via auto-oxidation of ferredoxin even in the presence of an oxygen trap (glucose and glucose oxidase). U nder optimum conditions of ferredoxin concentration the leak can be m inimized such th at H 2 production is about 70% of the ferricyanide Hill reaction rate.

The in vitro exchange between the epoxide oxygen of carotenoids and that of water reported in literature could not be verified. Our observations rather suggest that oxidation products of the labile polyene system may invalidate 18 O enrichment data obtained by indirect analysis.

Five different strains of bacteria ultilizing antipyrine as sole source of carbon were isolated from soil. It was shown by morphological and physiological examinations, that the new isolates are closely related to strains selected with the herbicide chloridazon. A ll of these bacteria are charac­ terized by special features and cannot be classified according to Bergey’s Manual of Determinative Bacteriology. Part of the strains which were selected with antipyrine not only grow with antipyrine but also with chloridazon. The others cannot be grown on chloridazon. However, resting cells of the latter group convert chloridazon to its catechol derivative (5-amino-4-chloro-2 (2,3-dihydroxyphenyl) -3 (2H)-pyridazinone). In these bacteria a catechol-2,3-dioxygenase (catechol: oxygen 2,3-oxido-reductase, EC 1.13.11.2) was found which readily catalyzes the cleavage of the catechol derivative of antipyrine (2,3-dimethyl-l-(2,3-dihydroxyphenyl)-pyrazolone (5)). The enzyme shows only slight activity with the corresponding derivative of chloridazon.

Further investigations on the structure of Friend murine leukemia virus (FLV) revealed that the transition from the “immature” (now termed “native”) to the structurally less organized “mature” (now termed “collapsed”) form occurs mainly as a result of the preparation for the electron microscope. A short pretreatment of virus with the detergent NP40 prior to negative staining with uranyl acetate is able to preserve the “native” structure of a high percentage of virus particles from standard preparations. Treatment with conventional fixatives was found to be in ­ effective. Using this preparation technique, a more detailed electron microscopic analysis of the viral internal organization became possible. A thin layer designated “inner coat” was newly detected in close apposition to the viral membrane between the viral envelope and core. Removal of the unit membrane by more intensive detergent treatment suggests the existence of material extending be­ tween the viral surface knobs and the viral interior. The icosahedral core shell has an opening at one side and this opening matches with the hole in the apparently beehive-like arranged ribo-nucleoprotein (RNP) strand which represents the innermost structure. A comparative study of a series of representative mammalian C-type viruses with the same technique indicated a close similarity to Friend virus in fine structure, although differences in stability were observed. Based on these and earlier findings a model of the structure of mammalian C-type viruses is presented.

The variation in visual acuity with illumination has been studied by monitoring the extracellular response of direction sensitive motion detecting neurons to a drifting sine wave grating displayed upon an oscilloscope spreen. Acuity reaches a maximum value of 0.46 cycles/degree at luminances above 1.0 cd/m 2 and decreases gradually over a 3.8 log unit attenuation in intensity to a minimum value of 0.05 cycles/degree. The results have been compared with theoretical acuity curves for the coupound eye with various dark adaptation mechanisms. The analysis indicates that a major strategy of dark adaptation in the fly is a process involving intensity-dependent neural summation of signals from photoreceptors having different visual axes.

The essential oil of freshly distilled leaves of Rosmarinus officinalis L. was found to contain 3-octanone, a compound hitherto not reported to be present in this oil. Identification was established by comparing the gas chromatographic retention time and by mass spectrometry.

The 5-deoxyisoflavones, afrormosin and formononetin have been isolated from healthy leaves of Onobrychis viciifolia. In addition to these compounds, several other flavonoid (iso-liquiritigenin; liquiritigenin; garbanzol) and isoflavonoid (medicarpin; vestitol) derivatives including the new iso-flavanone, vestitone (7,2′-dihydroxy-4′-methoxyisoflavanone), are present in leaves inoculated with the fungus Helmintho-sporium carbonum.

The mechanism of the biological hydroxylation of the isopropenyl side-chain of the sesquiterpene capsidiol, to give 13-hydroxycapsidiol, has been investigated with 13 C-NMR techniques, using capsidiol biogenetically enriched with 13 C from [1,2- 13 C 2 ] acetate. The results indicate that neither an allylic rearrangement nor the formation of an epoxide intervene in the process.

An enhancement of the rate of hydrolysis of certain methyl esters by cooperative effects of neighbouring amide and SH groups is demonstrated. The rate of hydroxyl ion catalyzed hydrolysis of the most reactive ester exceeds the rate of saponification of simple aliphatic esters by a factor of about 10 3 . A mechanism of the saponification reactions involving intermediate cyclic imide formation is proposed.

Properties of single cells in the posterior lateral (PL) eyes of the jum ping spider Plexippus validus are described. Only one spectral class of cells was encountered and the spectra sensitivity was indistinguishable from that measured from the ERG , both peaking at ca. 535 nm. Angular sensi­ tivity (width of angular sensitivity function at the 50% level) averaged .89° ± .12°, the smallest value being .77°. Absolute sensitivity (reciprocal of the number of quanta of peak wavelength, on axis required to generate a 50% response) averaged 1.43 + 2 .5 X 1 0 -11 q -1 ,cm 2 ·s. A ll cells studied were sensitive to the plane of polarised light. The per­ formance of receptors in the PL eyes is compared with that of receptors in the compound eyes of diurnal insects. It is concluded that the single lens eye system of spiders is in ­ herently superior in design to the insect compound eye.

The results show that both the direction and the intensity of the geomagnetic field can be sensed within the chamber of a Zeiss Cytopherometer. This suggests that electromagnetically induced fluid streaming might play a role in the perception of the geomagnetic field by organisms, although it is not clear at present in which organ the perception occurs. A basic requirement for such an organ would be an anisotropy of cells or cellular structures. The nervous system with its parallel axons, or specific cells associated with the nervous system, could thus be possible locations of the sensitivity towards magnetic fields.