Fluorescence of 3-keto-steroids in aqueous solution

The physiologically important 3-keto-steroids are non-fluorescent or only weakly fluorescent in protic as well as in aprotic solvents. In contrast, the 4,6,8(14)-triene-3-one steroids are highly fluorescent in aqueous solution but they do not appreciably fluoresce in other solvents. Evidence is presented that the introduction of double bonds into the skeleton of the 3-keto-steroids leads to a decrease of the energy of the lowest – ^* state, bringing this level into the neighbourhood of the non-fluorescent n – ^* state. As a consequence, for two states of approximately the same energy, relatively small perturbations such as those due to solvent interactions, protein binding and micelle formation, will then determine whether a system will fluoresce ( – ^* state lowest) or not (n – ^* state lowest). When the fluorescent 3-keto-steroids, having three conjugated double bonds, bind to proteins, the fluorescence intensity becomes almost zero, making these compounds useful as probes for steroid-protein interactions. This quenching of the fluorescence is explained by a decrease in energy of the n – ^* state relative to the – ^* state of the steroids due to hydrophobic interactions with the proteins.Abbreviations 6,8-BDT 6,8-bisdehydrotestosterone; DMSO, dimethylsulfoxide - HPLC high pressure liquid chromatography This work was presented in part at the Annual Meeting of the Gesellschaft für Biologische Chemie, September 26–29, 1983, in Göttingen. For an abstract see: Hoppe-Seyler's Z. Physiol. Chem. (1983) 364: 1151–1152Dedicated to Prof. Dr. F.-W. Zilliken on the occasion of his 65th birthday     

Vibronic attraction energy for superconductivity in the model of Si-containing organic polymers

Superconducting vibronic interaction has been examined for a model of Si-substituted polyacetylene. Large attractive vibronic interaction for the formation of superconducting electron pair has been found for the electron scattering process in between the and in-plane orbitals in silene unit. Doping of holes in these orbitals would be done by extracting electrons from the out-of-plane orbitals as well as from the in-plane orbitals. If stacking of the model polymers is performed in a parallel manner, the interplane electron pairing has also been found to be favorable on account of small Coulombic repulsion.     

Anionradicals from 5-halouracil substituted oriented DNA after X-irradiation at low temperatures

Conclusions The model of two initial radiation - induced radical components in oriented fibers of DNA,T ^– andG ⁺ [1], should be amended, for 5-halouracil substituted DNA by the finding that the primary anion is formed on the 5-halouracil base specifically but differentiates, depending on the halogen substituent, into*- and*-forms. The occurrence of the latter appears, from theoretical calculations, to be connected with a C-halogen bond elongation. The DNA matrix apparently allows for this whereas in the corresponding single crystals of 5-bromodeoxyuridine [1] or 5-iododeoxyuridine [4] the crystal packing prevents the elongation thus allowing for*-anion formation only.     

An underlying cause of measurement discrepancies in determinations of osmotic characteristics in plant cells and tissues

Summary It is frequently observed that liquid-exchange methods of measuring tissue water potential,, yield estimates further from zero than those obtained with vapour exchange methods, and that a similar situation exists with respect to plasmolytic methods of measuring vacuolar osmotic pressure,, compared with estimates from expressed sap methods. Although a number of factors may contribute to this phenomenon, it is suggested that the underlying reason is one of membrane leakiness, in the liquid exchange methods, to the solutes commonly employed. Although the amount of solute uptake may appear to be small during the period of the determinations, the effective osmotic pressure of the external solution is reduced if there is a net volume exchange of water and solute, instead of a simple water exchange. Thus the solution which appears to be in equilibrium with the tissue is, in fact, more concentrated than the true equilibrium solution leading to a spurious estimate of or .     

Osmotic potential and turgor maintenance in Spartina alterniflora Loisel.

Summary The dependence of leaf water potential (), osmotic potential () and turgor pressure (P) on relative water content (RWC) was determined for leaves of tall and short growth forms of Spartina alterniflora Loisel. from a site on Canary Creek marsh in Lewes, Delaware. Tall plants (ca. 1.5 m) occured along a drainage ditch where interstitial water salinity was approximately 20, and short plants (ca. 0.2 m) were 13 m away near a pan and exposed to 80 salinity during the most stressful period. Leaves were collected at dawn and pressure-volume measurements were made as they desiccated in the laboratory. Pressure equilibrium was used to measure , RWC was determined from weight loss and dry weight, was determined from the pressure volume curve, and P was calculated as the difference between and . Physical properties of the bulk leaf tissue that have a role in regulating water balance of the two growth forms were estimated: relative water content of apoplastic water (RWC_a) relative water content at zero turgor (RWC₀), the bulk modulus of elasticity (E), and water capacity (C _w). There were no detectable temporal trends in any of the parameters measured from Nune through September and no significant differences between the two growth forms when compared on the basis of RWC_a, RWC₀, E, and C _w. There was a clear difference between the two growth forms with respect to ; at RWC₀, was-4.5±0.40 MPa for short form plants and-3.3±0.40 MPa for tall form.Turgor pressure of plants in the field (P) was lower in leaves from short form than for the tall form plants with average difference of about 0.4 MPa. In July, P in short form leaves dropped to zero by mid-morning as expected for leaves experiencing water stress.These results show that S. alterniflora is capable of reducing osmotic potential in response to increased salinity and that turgor pressure was lower in short growth form than in tall forms.     

Factors influencing the isotopically exchangeable phosphate in soils

Summary The labile phosphate in a non-calcareous and a slightly calcareous soil was determined by isotopic exchange in the presence and absence of 0.001 molar solutions of a chelating and a non-chelating organic anion. The rate of isotopic exchange curves were analyzed graphically to sub-divide the labile phosphate into 3 or 4 fractions. The half-lives of exchange for the rapid, medium and slow fractions were between 0.3 to 1.6 hours, 1.8 to 8.6 hours and 25.8 to 46.1 hours respectively. In addition, an instantaneously-exchanging component was sometimes observed.In the presence of the citrate ion, the total labile phosphate was increased in the non-calcareous soil and decreased in the calcareous soil, whereas the diethyl barbiturate ion (DEB) anions decreased the total labile phosphate in both soils. In general, the citrate ion increased the rapid and the medium fractions whereas the DEB anion either did not affect them or decreased them. Again, whereas citrate always increased the phosphate in solution, the effect of DEB anions depended on the soil. The major effect of the organic anions was to greatly decrease the slowly-exchanging fraction in both soils.The half-lives of exchange for the rapid, medium and slow fractions were in the order no organic anion > citrate > barbiturate and the rate constants for a first-order mechanism were in the order no organic anion < barbiturate < citrate.Small but significant differences were observed between the two soils.     

A novel pollen-specific α-tubulin in sunflower: structure and characterization

We describe here a new -tubulin isoform from sunflower we named -tubulin. -tubulin is the most divergent higher-plant -tubulin described so far, having an unusual deletion in the H1/B2 loop and a glutamine-rich C-terminus. We constructed a three-dimensional model and discuss its implications. Using specific antibodies, we show that -tubulin expression is restricted to the male gametophyte. -tubulin mRNA represents 90% of -tubulin mRNA and a small percentage of total pollen mRNA. Among the plants tested, -tubulin was only detected in sunflower and in Cosmos. Since both plants are Asteraceae, we propose that -tubulin is specific to this family. Our results suggest that -tubulin can inhibit tubulin assembly in pollen. This hypothesis is reinforced by the fact that -tubulin is found in a complex with -tubulin in mature sunflower pollen.     

DNA polymorphisms associated with a new α1-antitrypsin PI Q0 variant (PI Q0riedenburg)

Summary A new PI Q0 variant (PI Q0_riedenburg) is described; it is caused by a complete deletion of the ₁-antitrypsin (₁AT) gene. The deletion gives rise to four new restriction fragment length polymorphisms (RFLPs) detected with a genomic probe of the 5 region of the gene. Analysis of the RFLPs indicates that the deletion starts immediately upstream of exon Ic. The deletion extends into the 3 flanking region of the gene but does not include the ₁AT-related gene (the PIL gene), which is located 12 kb downstream of the ₁AT gene.     

Does glutathione S-transferase Pi (GST-Pi) a marker protein for cancer?

Glutathione S-transferases (GSTs, EC 2.5.1.18) are multifunctional and multigene products. They are versatile enzymes and participate in the nucleophilic attack of the sulphur atom of glutathione on the electrophilic centers of various endogenous and xenobiotic compounds. Out of the five, , and , major classes of GSTs, GST- has significance in the diagnosis of cancers as it is expressed abundantly in tumor cells. This protein is a single gene product, coded by seven exons, that is having 24 kDa mass and pI value of 7.0. Four upstream elements such as two enhancers, and one of each of AP-1 site and GC box regulate gene. During chemical carcinogenesis because of jun/fos oncogenes (AP-1) regulatory elements, specifically GST- is expressed in liver. Therefore this gene product could be used as marker protein for the detection of chemical toxicity and carcinogenesis.     

The role of β-dimethylsulphoniopropionate,glycine betaine and homarine in the osmoacclimation of Platymonas subcordiformis

The tertiary sulphonium compound, -dimethylsulphoniopropionate (DMSP) and the quaternary ammonium compounds glycine betaine and homarine are important osmotica in Platymonas subcordiformis cells. Following hypersaline stresses the compounds were accumulated after a lag period of 3 h and equilibrium concentrations were reached 6 h later. In contrast to these organic solutes, mannitol was synthesised immediately and equilibrium concentrations were reached within 90 min. Hyposaline stresses induced losses of the organic solutes from the cells. The ions K⁺, Na⁺, Cl^- and the above organic solutes can account for the osmotic balance of the cells.Abbreviations DMSP -dimethylsulphoniopropionate - _i intracellular osmolality - _o extracellular osmolality     

Hämatologische Beobachtungen an Regenbogenforellen (Salmo gairdneri RICH.). V. Die Erythozytenverteilungskurve

Zusammenfassung Für die Fischpathologie wird als differentialdiagnostisches Verfahren die Erythozytenmessung vorgeschlagen. Zur Darstellung der Erythrozytenverteilungskurve wird wegen der elliptischen Form der Fisch-Erythrozyten die in der Humanmedizin übliche Technik modifiziert.An 102 Regenbogenforellen (Salmo gairdneri) beiderlei Geschlechts im Alter von 1 bis 3 Jahren wurde versucht, die Normalkurve für gesunde Tiere dieser Population zu ermitteln. Die Größenverteilung der Erythrozyten ergibt eine Gauss-Glockenkurve, deren Gipfel in die Größenklasse 30–35 µm²/ fällt, und deren Basis von 15–20 µm²/ bis 55–60 µm²/ reicht.Zwischen den Price-Jones-Kurven der Milchner (52 Individuen) und Rogner (50 Individuen) bestehen keine signifikanten Unterschiede. Ebenso konnte keine Beeinflussung dieser Normalkurve durch unterschiedliches Alter oder jahreszeitliche Einflüsse — untersucht wurden in dieser Hinsicht die F₂ — beobachtet werden.

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Summary As differential-diagnostic feature in the fish pathology it is suggested to measure the red blood cells. The normal technique used in human medicine for plotting the distribution curve of the red blood cells is modified, because the red blood cells of the fish are of elliptical shape.102 rainbow trouts (Salmo gairdneri), both, male and female, and 1–3 years of age, were examined to find the normal curve for healthy animals of this population. The size distribution of the red blood cells show a bell-shaped Gauss-curve, the peak of which is 30–35 µm²/, and the basis of which reaches from 15–20 µm²/ to 55–60 µm²/.The Price-Jones-curves show no significant differences between the milters (52 individuals) and the spawners (50 individuals). Besides, changes caused by different age or seasonal influences could not be observed on this normal curve. Analyzed were the F₂ individuals.

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Institut für Siedlungswasserbau und Wassergütewirtschaft der Universität Stuttgart Biologische Abteilung - Arbeitsgruppe Haider     

Identification of a glutathione-S-transferase effector domain for inhibition of jun kinase, by molecular dynamics

We have recently found that the glutathione-S-transferase -isozyme (GST-), a cellular detoxification enzyme, potently and selectively inhibits activation of jun protein by its upstream kinase, jun kinase (JNK). This newly identified regulatory activity of GST- is strongly inhibited by a group of agents that inhibit its enzymatic activity. Since loss of enzymatic activity in general does not correlate with loss of regulatory activity, it is likely that inhibitor binding induces changes in the structure of one or more domains of GST that block its interaction with JNK. To identify regions of GST that change conformation on the binding of inhibitors, we have performed molecular dynamics calculations on GST- to compute its average structure in the presence and absence of the inhibitor, glutathione sulfonate. Superposition of the two average structures reveals that several regions change local structure depending upon whether the inhibitor is bound or not bound. Two of these regions, residues 36–50 and 194–201, are highly exposed. We have synthesized peptides corresponding to these two segments and find that the 194–201 sequence strongly inhibits the ability of GST- to block the in vitro phosphorylation of jun by JNK. These results suggest that this region of GST- is critical to its functioning as a newly discovered regulator of signal transduction.     

Expression in Saccharomyces cerevisiae of a gene associated with cytoplasmic male sterility from maize: Respiratory dysfunction and uncoupling of yeast mitochondria

Summary The replication initiation protein of the Escherichia coli plasmid R6K is a dual regulator in the control of plasmid copy number, functioning both as a specific initiator and inhibitor of replication. While the biochemical basis of these activities is not known, initiator activity requires binding of the protein to the seven 22 by direct repeats within the -origin region. By deleting C-terminal segments of the coding region, we have found that the N-terminal polypeptides of that are produced, corresponding to the first 117 and 164 amino acids, respectively, retain the negative activity of the bifunctional protein, i.e. these truncated proteins specifically inhibit R6K replication in vivo. These negatively acting polypeptides, however, are incapable of initiating replication in vivo and fail to bind to the -origin of the R6K DNA in vitro. A correspondence between the observed negative activity of the N-terminal peptide and the negative regulatory activity of the intact protein is supported by the finding that point mutations introduced into the 164 amino acid N-terminal peptide that result in a decrease in its inhibitory activity also produce a plasmid high-copy phenotype when these mutations are incorporated into the full-length protein. These findings demonstrate that the negative domain of resides in the N-terminal segment of the protein. Furthermore, the data obtained suggest that inhibition of R6K replication by does not require direct binding to DNA.     

Photosynthesis of isolated chloroplasts and protoplasts under osmotic stress

1. Isolated intact spinach chloroplasts respond to changes of the sorbitol concentration of the suspending medium as near-perfect osmometers within a large range of osmotic potentials. Under isotonic conditions (=9–10 bar), their average osmotic volume is 24 m³ and the total volume 36 m³. The osmotic volume can be increased to 63 m³ by lowering the sorbitol concentration until a critical osmotic potential of =4 bar is reached. Below that value chloroplasts rupture. Between 10 bar and 4 bar, volume changes are reversible. 2. Increasing the chloroplast volume above 24 m³ causes inhibition of photosynthesis, with 50% inhibition occurring at an osmotic potential of =5–6 bar. This corresponds to an osmotic volume of 45–55 m³. Depending on the duration of hypotonic treatment, inhibition of photosynthesis is more or less reversible. 3. Between 4 and 10 bar, the chloroplast envelope exhibits a very low permeability for ferricyanide, many metabolites, and soluble stroma proteins. 4. Electron transport is not inhibited by swelling of chloroplasts. Also, the ATP/ADP-ratio remains unchanged. 5. The solute concentration in the chloroplasts appears to be optimal for photosynthesis at 10 bar. Increasing the chloroplast volume causes inhibition of photosynthesis by dilution effects.     

Electronic structures of donor-acceptor polymers based on polythiophene,polyfuran and polypyrrole

Theoretical results on the geometric and electronic structures of some donor-acceptor polymers based on polythiophene (X=S), polyfuran (X=O) and polypyrrole (X=NH) were obtained, using a one-dimensional tight-binding self-consistent field crystal-orbital (SCF-CO) method at the MNDO-AM1 level of approximation. The repeat unit of these polymers consits of a bithiophene, furan or bipyrrole unit bridged by an electron-accepting group or. The optimized geometries of the polymers show a strong dependence on the nature of the electron donating group X. All the polymers studied are predicted to have band gap values ranging between 1 eV and 2 eV. An analysis of their -bond order data and of the patterns of their frontier orbitals shows they have benzenoid-like electronic structures.     

Identification and characterization of the functional alpha origin of DNA replication of the R6K plasmid and its relatedness to the R6K beta and gamma origins

Summary The functional R6K origin is composed of two DNA elements, one of 580 bp carrying the origin sequences and the other of 277 bp containing the seven 22 bp direct repeats previosly identified as also required for and origin activity. These two genetic elements are separated by approximately 3,000 bp of R6K sequences which are dispensable for origin activity. The function of the origin depends on the presence in cis of the 580 bp and the 277 bp fragments and requires that they be oriented as in the intact R6K. Activation of the origin depends on the R6K replication initiation protein .Within the 580 bp of the origin, there is a sequence of 98 bp which appears as an inverted repeat of 96 bp in the replicon. Deletion of the 96 bp or 98 bp results in inactivation of the and the origins respectively. These long repeats are palindromic and it is suggested that these may serve as the recognition signals for initiation of DNA replication in the and the origins of R6K. DNA homology analysis performed on , and origin sequences, also reveals 10–23 bp sequences in the and the origins that are related to the family of 22 bp direct repeats in the origin which were shown previously to be binding sites for the protein.     

Correlation between changes in photosynthetic activity and changes in total protoplast volume in leaf tissue from hygro-, meso- and xerophytes under osmotic stress

Rates of photosynthesis of leaf slices from various hygro-, meso- and xerophytes were measured in the absence of stomatal control in various stages of osmotic dehydration. The external osmotic potential _° for a 50% inhibition of photosynthesis varied between 20 bar in some hygrophytes up to 50 bar in xerophytes. The response of photosynthetic enzymes to increased salt concentrations in the reaction medium was similar in leaf extracts from hygro-, meso- and xerophytes. The total protoplast volume in vacuum-infiltrated leaf discs from various plants was measured as the difference between ³H₂O-labeled space and [¹⁴C]sorbitol-labeled space. In all plants, the protoplast volume could be reduced to about 55% of the maximum volume of tissue in equilibrium with water, without decreasing photosynthesis. Reduction of the maximal protoplast volume below 55% decreased photosynthesis in all tissues to the same decreased photosynthesis in all tissues to the same degree. At 20% maximal volume, photosynthesis of all plants was completely inhibited. The differential decrease of protoplast volumes of various leaf tissues in response to changes in _° was mainly due to the different osmotic potential of the cell sap (_cs). The relative contribution of sugars to the overall osmolarity of the cell sap was up to nineteen times higher in xerophytes than in hygrophytes. Short-term recovery of photosynthesis after hypertonic stress was good in xerophytes, incomplete in mesophytes and absent in hygrophytes. There was also a large discrepancy between the partial recovery of protoplast volumes and the complete absence of a recovery of photosynthesis in hygrophytes.     

Chloroplast acclimation to low osmotic potential

Photosynthetic potential of isolated chloroplasts was investigated during in situ water deficits. An eight day stress cycle imposed on spinach plants reduced leaf _w by 0.57MPa, and leaf by 0.50MPa, resulting in partial turgor maintenance during the stress cycle. Pressure/volume curves confirmed the occurrence of osmotic adjustment. Leaf depression was associated with an altered response of chloroplasts to low in vitro. Optimum reaction medium for photosynthesis shifted from –1.04 to –1.57MPa, and low was not as inhibitory to photosynthesis of plastids pre-exposed to stress in situ. These data indicate that chloroplasts acclimate to low external in response to leaf water deficits. This response was still evident four days after a stress cycle ended, but was nearly reversed eight days after stress. Repeated stress cycles in situ did not increase the degree of chloroplast acclimation to low in vitro. Fast dehydration of leaves did not induce this apparent chloroplast acclimation.Abbreviations osmotic potential - _w water potential - PEG polyethylene glycol 8000 - MPa megapascals     

Syntheses of α- and β-Glycosyl Donors with a Disaccharide β-D-Gal-(1→3)-D-GalNAc Backbone

The synthesis of thioglycoside glycosyl donors with a disaccharide -D-Gal-(1 3)-D-GalNAc backbone was studied using the glycosylation of a series of suitably protected 3-monohydroxy- and 3,4-dihydroxyderivatives of phenyl 2-azido-2-deoxy-1-thio-- and 1-thio--D-galactopyranosides by galactosyl bromide, fluoride, and trichloroacetimidate. In the reaction with the monohydroxylated glycosyl acceptor, the process of intermolecular transfer of thiophenyl group from the glycosyl acceptor onto the cation formed from the molecule of glycosyl donor dominated. When glycosylating 3,4-diol under the same conditions, the product of the thiophenyl group transfer dominated or the undesired (1 4), rather than (1 3)-linked, disaccharide product formed. The aglycon transfer was excluded when 4-nitrophenylthio group was substituted for phenylthio group in the galactosyl acceptor molecule. This led to the target disaccharide, 4-nitrophenyl 2-azido-4,6-O-benzylidene-2-deoxy-3-O-(2,3,4,6-tetra-O-acetyl--D-galactopyranosyl)-1-thio--D-galactopyranoside, in 57% yield. This disaccharide product bears nonparticipating azido group in position 2 of galactosamine and can hence be used to form -glycoside bond. Azido group and the aglycon nitro group were simultaneously reduced in this product and then trichloroacetylated, which led to the -glycosyl donor, 4-trichloroacetamidophenyl 4,6-di-O-acetyl-2-deoxy-3-O-(2,3,4,6-tetra-O-acetyl--D-galactopyranosyl)-1-thio-2-trichloroacetamido--D-galactopyranoside, in 62% yield. The resulting glycosyl donor was used in the synthesis of tetrasaccharide asialo-GM₁.     

A method for the investigation of those transformations under which the visual recognition of a given object is invariant

An experiment is described which shows in operation the program set out in Foster (1972a) for the investigation of the invariance transformations of visual recognition. The concern in the present study is with the Lie group of rotations SO(2), and a certain centrally located foveal Landolt ring. By presenting to the visual system this Landolt ring and a rotated image in rapid succession, one attempted to induce a specified rotation-type phi-motion. Two subjects were employed. Both reported the existence of the required type of phi-motion for rotations ₀ of the Landolt ring about the visual axis with -2/72/7. By appealing to the basic Proposition 2 of Foster (1972 a), the conclusion is reached that the visual system appears capable of effecting upon a certain centrally located foveal annulus the local 1-parameter group of rotations about the visual axis ₀, [–2/7,2/7].