AT-1 Receptor and Phospholipase C Are Involved in Angiotensin III Modulation of Hypothalamic Noradrenergic Transmission

SUMMARY 1. We previously reported that angiotensin III modulates noradrenergic neurotransmission in the hypothalamus of the rat. In the present work we studied the effects of angiotensin III on norepinephrine release and tyrosine hydroxylase activity. We also investigated the receptors and intracellular pathways involved in angiotensin III modulation of noradrenergic transmission.2. In rat hypothalamic tissue labeled with [³H]norepinephrine 1, 10, and 100 nM and 1 M losartan (AT1 receptor antagonist) had no effect on basal neuronal norepinephrine release, whereas 10 and 100 nM and 1 M losartan partially diminished norepinephrine secretion evoked by 25 mM KCl. The AT2 receptor antagonist PD 123319 showed no effect either on basal or evoked norepinephrine release. The increase in both basal and evoked norepinephrine output induced by 1 M angiotensin III was blocked by 1 M losartan, but not by 1 M PD 123319.3. The phospholipase C inhibitor 5 M neomicin inhibited the increase in basal and evoked norepinephrine release produced by 1 M angiotensin III.4. Tyrosine hydroxylase activity was increased by 1 M angiotensin III and this effect was blocked by 1 M LST and 5 M neomicin, but not by PD 123319. On the other hand, 1 M angiotensin III enhanced phosphatidyl inositol hydrolysis that was blocked by 1 M losartan and 5 M neomicin. PD 123319 (1 M) did not affect ANG III-induced phosphatidyl inositol hydrolysis enhancement.5. Our results confirm that angiotensin III acts as a modulator of noradrenergic transmission at the hypothalamic level through the AT1-phospholipase C pathway. This enhancement of hypothalamic noradrenergic activity suggests that angiotensin III may act as a central modulator of several biological processes regulated at this level by catecholamines, such as cardiovascular, endocrine, and autonomic functions as well as water and saline homeostasis.     

Carotenoids in the eyespot apparatus of the flagellate green alga Spermatozopsis similis: Adaptation to the retinal-based photoreceptor

Isolated intact eyespot apparatuses, the photoreceptive organelles involved in blue-light-mediated photoresponses of flagellate green algae, were analyzed regarding their carotenoid composition. Carotenoids from the eyespot apparatuses of Spermatozopsis similis were identified by high-performance liquid chromatography, visible-light absorption spectra, mass spectroscopy and by ¹H-nuclear magnetic resonance spectroscopy (carotenes), and compared with those of whole-cell extracts. Both extracts contained ,-carotene, ,-carotene (formerly -carotene), lycopene, lutein, zeaxanthin, violaxanthin and all-E-and 9-Z-neoxanthin. The relative carotenoid compositions, however, differed significantly. A twofold relative increase in the total carotene level was evident in the fraction enriched in eyespot apparatuses. This was mainly due to an increase in the monocyclic ,-carotene and the aliphatic lycopene, whereas the relative content of ,-carotene remained unchanged. On the other hand a relative decrease in the total xanthophyll content, especially of lutein and the epoxidic carotenoid neoxanthin, was observed in the eyespot apparatuses compared with the whole-cell extracts. The decrease of the latter resulted almost solely from a reduction of the 9-Z-rather than the all-E-isomer. The bulk of the carotenes is thought to be localized in the highly organized eyespot lipid globules, which act as a combined quarter-wave interference reflector and absorption screen for the photoreceptor in green algae. The enrichment of ,-carotene and lycopene in the eyespot apparatuses, extending the range of visible light absorption to longer wavelengths, represents an adaptation of the screen to the retinal-based photoreceptor of flagellate green algae and is one of the prerequisites for maximal directional sensitivity of the eyespot apparatus.Abbreviations ¹H-NMR nuclear magnetic resonance - IUPAC International Union of Pure and Applied Chemistry - VIS visible absorption spectra This work was supported by the Deutsche Forschungsgemeinschaft (G.K. and M.M.). M.G. was supported by a fellowship from the Norwegian Research Council of Science and Humanities.     

Über den Tentakelapparat der Edelkoralle (Corallium rubrum L.) und seine Funktion beim Beutefangverhalten

Zusammenfassung Die Tentakel der Edelkoralle vermögen sich an den Spitzen bis über die dreifache Länge der Normtentakel fadenartig zu verlängern (Fadententakel). Diese Tentakelfäden wickeln sich bei Berührung eines Nahrungsobjekts korkzieherartig auf und nähern dieses durch Kontraktion der Mundöffnung.Damit koordiniert führen die zugehörigen Basalabschnitte (Normtentakel) durch S-förmige Biegungen und eventuelle Verkürzungen die Stelle der gereizten Tentakelfadens präzise zum Mund.Werden die Normtentakel allein gereizt, so antworten sie mit raschem Einschlag zur Mundöffnung. Bei gleichzeitiger Reaktion aller Tentakel wird dadurch eine Tentakel-Faust oder ein Fangkorb gebildet. Bei schwachen Reizen antwortet der Normtentakel anstelle der schlagenden Reaktionen mit langsamen Krümmungen, die das Objekt zur Mundöffnung dirigieren. Unabhängig von der Reaktionsart der Tentakeln wird die Beute vor der Aufnahme stets geprüft und häufig abgelehnt. Ist der Polyp in Freßstimmung, so wird die Nahrung rasch durch das glockenartig erweiterte Schlundrohr geschleust und verschwindet nach wenigen Minuten im Coelenteron.

---

The tentacle apparatus of the red coral (Corallium rubrum L.) and its role in feeding behaviour

Summary The tentacles of the octocoral Corallium rubrum may become extremely elongated (threadlike-tentacle) the apical part of the norm-tentacle becoming stretched more than three times as much as the base.The threadlike parts of the tentacles contract corcscrew-like when touched by food particles.The stimulated part of the tentacle is brought close to the mouth by coordinated movements of its basal portion. Very often both threadlike-tentacle and norm-tentacle shorten in order to carry the particle to the pharynx.When the norm-tentacles only are touched, they react with quick strokes towards the oral disc. This behaviour, when shown by all tentacles at the same time results in the formation of either a tentacle-fist or a basketlike structure. At slight irritations the norm-tentacle will respond with slow bending movements, which lead the particle towards the mouth. Independent of the type of tentacle reactions the prey is always checked before entering the pharynx, and very often refused.If the polyp decides to accept, food is transported quickly through the elongated pharynx and within a few minutes will have disappeared in the coelenteron.

     

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.     

Regeneration of plantlets through organogenesis in the Himalayan yellow poppy,Meconopsis paniculata

Plantlet formation through organogenesis in callus cultures of Himalayan yellow poppy,Meconopsis paniculata D.Don (Prain), a threatened taxon of ornamental value, is described. Hypocotyl segments from 3-month-old laboratory-raised seedlings produced callus on agar-solidified Murashige and Skoog medium (MS) containing 10 M -naphthaleneacetic acid and 1 M kinetin. Shoots differentiated best from callus on MS containing 10 M indolebutyric acid (IBA) and 1 M 6-benzyladenine. The regenerated shoots rooted best on MS medium containing 10 M IBA. From seed germination to differentiation of plantlets through the two-step organogenesis process required 28–29 weeks.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - FAA formalin-acetic acid-alcohol - BA 6-benzyladenine - IAA indole-acetic acid - IBA indolebutyric acid - GA₃ gibberellic acid - NAA -naphthaleneacetic acid - RH relative humidity     

Relationship of amino acid sequence to immunological activity — syntheses and structure-activity relationships of thymosinβ 4 family

Summary Four peptides related to thymosin ₄ family and its six fragments were synthesized by the solution method. Among them, the four peptides related to thymosin ₄ family and its two fragments were found to have restoring activity on the impaired blastogenic response of T-lymphocytes isolated from uremic patients, but the other four fragments had no effect. Studies on the structureactivity relationships suggest that the tricosapeptide moiety corresponding to amino acids 16–38 of thymosin ₄ is found to be an important moiety on impaired immunological deficiency.Amino acids and their derivatives used in this investigations were of the L-configuration. The following abbreviations are used: DMF, dimethylformamide; DMSO, dimethylsulfoxide; Boc, tert-butoxycarbonyl; Z, benzyloxycarbonyl; NP, p-nitrophenyl; ONp, p-nitrophenyl ester; OBzl, benzyl ester; Bzl, benzyl; Troc,,,-tricloroethoxycarbonly; Su, N-hydroxysuccinimide; NMM, N-methylmorpholine; OSu, N-hydroxysuccinimide ester; WSCI, l-ethyl-3; (3-dimethylaminopropyl) carbodiimide; HOBT, N-hydroxybenzotriazole; TFA, trifluoroacetic acid; TLC, thin-layer chromatography; E-rosette, a rosette with sheep erythrocytes; EDTA, ethylenediaminetetraacetic acid; PHA, phytohemagglutinin; HPLC, high-performance liquid chromatography; ONb, p-nitrobenzyl ester; DCC, dicyclohexyl-carbodiimide.     

RFLP analysis of genomic regions associated with cooked-kernel elongation in rice

As a result of earlier breeding efforts, portions of the genome of Basmati 370 have been introgressed into a rice breeding line, B8462T3-710. Cooked-kernel elongation was increased in this breeding line to a level equal to that of Basmati 370. The objective of this study was to identify and locate quantitative trait loci (QTLs) associated with cooked-kernel elongation in an F3 population derived from a cross between B8462T3710 and the reduced-elongation recurrent parent variety, Dellmont. DNA from the parental lines and Basmati 370 as a control, were screened for RFLPs using 170 clones chosen to cover the rice genome at intervals of 8 cM on average. Eighteen markers identified RFLPs common to Basmati 370 and B8462T3-710, but different from Dellmont, suggesting possible associations with kernel elongation. The B8462T3-710/Dellmont F3 population was analyzed for segregation of those RFLPs and for kernel elongation. Analysis of variance of the kernel elongation ratio revealed that two markers, 14.6 cM apart on chromosome 8, are significantly associated with this trait (RZ323 P 0.005, RZ562 P 0.05). Interval mapping suggests a single QTL with a close proximity to RZ323. This QTL was tested in F6 lines derived from the same cross and the presence of the B8462T3-710 segment detected by RZ323 caused a highly significant increase of the kernel elongation ratio (P 0.04). In addition, the QTL for kernel elongation and a gene for aroma, which are major components of the grain quality characteristics of Basmati-type rices, showed linkage. The availability of linked markers to the QTL may facilitate early selection for kernel elongation in rice breeding programs.     

Transformation of lithocholic acid to a new bile acid, 3α, 15β-dihydroxy-5β-cholanic acid by Cunninghamella blakesleeana ST-22

Summary A fungus identified as Cunninghamella blakesleeana (Lendner) can carry out 15-hydroxylation of lithocholic acid to a new bile acid (3,15-dihydroxy-5-cholanic acid). By optimizing the fermentation conditions, the amount of the product increased from 0.17 g/l to 1.2 g/l. Hydrophilicity measurements and in vitro cholesterol solubilization tests showed that 3, 15-dihydroxy-5-cholanic acid was as effective as ursodeoxycholic acid in cholesterol solubilization.Abbreviations LCA lithocholic acid (3-hydroxy-5-cholanic acid) - 3, 15-DHC (3, 15-dihydroxy-5-cholanic acid) - DMSO dimethyl sulfoxide - CHES 2-[N-cyclohexylamino]ethanesulfonic acid     

A quick and efficient system for antibiotic-free expression of heterologous genes in tobacco roots

Requirement for antibiotic-resistance selection markers and difficulty in identifying transgenes with the highest expression levels remain the major obstacles for rapid production of recombinant proteins in plants. An alternative approach to producing transgenic plants free of antibiotic-resistance markers is the phenotypic-based selection with root-proliferation genes (rol genes) of Agrobacterium rhizogenes. By using Agrobacterium tumefaciens harboring the pRYG transformation vector with a cluster of rol genes linked to a heterologous gene of interest, we have developed a rapid transformation tool using hairy root formation as a selection marker. The expression of -glucuronidase in newly induced transgenic tobacco roots could be detected as early as 12 days after inoculation. Higher levels of transgene expression in the roots correlated positively with the rates of root elongation on hormone-free medium and thus could be used for positive selection. When tobacco plants were transformed with pRYG harboring the expression cassette for secreted alkaline phosphatase (SEAP), the release of SEAP from roots of the fully regenerated transgenic plants could be quantified at rates as high as 28 g/g root dry weight per day.Abbreviations GUS: -Glucuronidase - SEAP: Secreted alkaline phosphatase - rolABC: Cluster of rolA, rolB, and rolC genesCommunicated by A. Altman     

Association between defects of karyogamy and translation termination in yeast <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis>

Mutants capable of a high frequency of cytoduction (Hfc⁺) were obtained in a haploid strain of Saccharomyces cerevisiae, suggesting impaired cytogamy. Nine of the 68 Hfc⁺ mutants showed the antisuppressor effect with respect to mutations of the SUP35 and SUP45 genes, which code for translation termination factors, or to the [PSI ⁺] factor, which is the prion form of Sup35. Cosegregation of the characters higher frequency of cytoduction and antisuppression was demonstrated for three Hfc⁺ mutants. One (HFC12-2) of the Hfc⁺ mutations exerted a dominant antisuppressor effect with respect to [PSI ⁺] and had no effect on [PSI ⁺] maintenance. On the strength of the results, an interaction was assumed for translation termination components and cytoskeleton proteins, which play a role in karyogamy in yeasts.Translated from Genetika, Vol. 41, No. 2, 2005, pp. 178–186.Original Russian Text Copyright © 2005 by Borchsenius, Repnevskaya, Kurischko, Inge-Vechtomov.     

Optimal and central-place foraging theory applied to a desert harvester ant,Pogonomyrmex californicus

Summary Certain predictions of optimal- and central place-foraging theory were tested on the desert harvester ant, Pogonomyrmex californicus. Colonies were offered three different sizes of oat seed and found to maximize net energy intake (ei) over time (t _i ) by harvesting the seed sizes with the highest e _i /t _i rank. Two aspects of t _i were measured that were assumed constant in previous studies. The handling components of t _i (time required to manipulate the seed and travel time back to the colony with the food) were measured and found to be positively correlated with seed size. The manipulation success rate (the percentage of handled seeds successfully picked up) decreased with increased seed size. These results point out how important it is to measure all parameters of e _i /t _i rather than to assume constancy with both prey type and foraging distance. The relative abundance of less preferred food types was important in determining the proportion of preferred types in the diet. The food supply of eight colonies was manipulated experimentally over a 25-day period. Four deprived colonies were constrained within aluminum enclosures to prevented foraging. The remaining four satiated colonies were given food ad libitum. The niche breadths of the treated colonies were then compared to controls, but found not to differ significantly. Seed baits were offered at three distances from the colony to test whether selectivity increased with disance. Contrary to theoretical predictions, all colonies harcested about the same proportion of each seed size at each distance.     

Mechanism of food detection in the tadpoles of the bronze frog <Emphasis Type="Italic">Rana temporalis</Emphasis>

Mechanism of food detection in Rana temporalis tadpoles was studied using a rectangular choice tank with end compartments (stimulus zones) providing exclusively visual and/or chemical food cues. Boiled spinach served as the food. The test tadpoles were starved for 24 h before use. They were released from the center of the choice tank (n=24) after 5 min of acclimation to test for end bias and food-detecting mechanism. The number of tadpoles in the two stimulus zones was recorded at 5-min intervals from 10 to 30 min. In the end-bias tests (without food cues) tadpole distribution was comparable at all times in the two compartments of the choice tank, exhibiting no end bias. In tests with the visual food cues provided in one of the stimulus zones, the tadpole distribution was also random. On the other hand, in experiments involving chemical cues emanating from food the tadpoles preferentially associated with the food source in significantly greater numbers compared to the zone lacking food or providing only a visual cue. The experiments with individual test tadpoles also revealed that they detect food based on chemical cues and ruled out copycat behavior. These findings on R. temporalis tadpoles reveal that chemical senses predominate over the visual senses in detection of food and foraging.     

Hydrogenase of purple bacteria: properties and regulation of synthesis

Physico-chemical properties of homogeneous preparations of soluble and membrane-bound hydrogenases from the purple sulfur bacterium Thiocapsa roseopersicina BBS and membrane-bound hydrogenase of Rhodopseudomonas capsulata, strain B10 have been studied. Compared to the enzymes from other sources, the hydrogenase of Thiocapsa roseopersicina is more stable to O₂ and products of its reduction (O ₂ ^- , H₂O₂), temperature and a number of other factors of the medium. A natural electron donor for T. roseopersicina hydrogenase is a low-potential cytochrome C₃, while the natural electron acceptors for hydrogenases of R. capsulata, T. roseopersicina, Ectothiorhodospira shaposhnikovii and Anabaena cylindrica are cytochromes of groups c and b.In different phototrophs, synthesis of hydrogenase can be inhibited by the presence of high concentrations of O₂. In some microorganisms (e.g. Rhodopseudomonas capsulata, strain B10) the repressing effect on hydrogenase formation is also exhibited by organic compounds. H₂ may not necessarily be present for hydrogenase synthesis by purple bacteria, but its presence may considerable increase the level of the enzyme.Abbreviations SDS sodium dodecylsulfate - Hipip high-potential iron — sulfur protein - R Rhodopseudomonas - T Thiocapsa - Rh Rhodospirillum - C Chromatium This paper is dedicated to Professor Dr. H.G. Schlegel in honour of his sixtieth birthday and in recognition of his great contribution in the field of physiology and biochemistry of microorganisms capable of using H₂. Professor H.G. Schlegel had a profound and most fuitful influence on the progress in the research of the laboratory headed by the author     

A new approach to the immunocytochemistry of cAMP

A method is described to couple cyclic adenosine 3,5 monophosphate (cAMP) to a carrier protein by means of acrolein. Antibodies against this conjugate were raised in mice. These antibodies proved to be highly specific for acrolein-fixed cAMP in a gelatin model system. Slices (300 m in thickness) from rat cerebral cortex were incubated in vitro and the dopaminergic control of adenylate cyclase activity was drug-manipulated. This manipulation was visualized by application of the cAMP-antisera on cryostat sections of the acrolein fixed slices.     

Growth retarding effects of the herbicide difenzoquat

The growth-regulating properties of the herbicide Difenzoquat (1,2-dimethyl-3,5-diphenyl-1-H-pyrazolium methyl sulfate) were investigated in seedlings and cell suspension cultures of sunflower (Helianthus annuus L.). Application of 10 g or more Difenzoquat to the apex of seedlings resulted in a transient inhibition of internode elongation. Application of GA₃ to treated seedlings resulted in enhanced internode elongation but did not reverse the degree of growth inhibition elicited by Difenzoquat. Endogenous gibberellin levels were estimated by bioassay and were qualitatively and quantitatively similar in extracts from control and treated seedlings. Treatment of suspension cultures of sunflower cells with 1 M or more Difenzoquat resulted in an inhibition of cell division (dry-matter accumulation). Neither GA₃ nor a mixture of sterols (cholesterol, -sitosterol, and stigmasterol) alone or in combination was able to overcome this inhibition of cell division. It was concluded that the growth-retarding activity of Difenzoquat was the result of its action at the cellular level and was not mediated by inhibition of gibberellin biosynthesis.     

Transformation of haploid, microspore-derived cell suspension protoplasts of rice (Oryza sativa L.)

We compared the transient activity of three cereal gene-derived promoter-gus fusions and the efficiency of selection mediated by three different selectable genes in a polyethylene glycol transformation system with haploid cell suspension protoplasts of rice. The maize ubiquitin promoter was found to be the most active in transformed protoplasts, and selection on ammonium glufosinate mediated by the bar gene was the most efficient for producing resistant calluses. Cotransformation of protoplasts with two separate plasmids carrying the gus and the bar genes, at either a 21 or 11 ratio, led to 0.8 × 10^–5 and 1.6 × 10^–5 resistant callus recovery frequencies and 59.7 and 37.9 cotransformation efficiencies respectively. No escapes were detected in dot blot analyses of 100 resistant calluses with a probe consisting of the bar coding region. Cotransformation efficiency, based on resistance to basta and -glucuronidase staining of the leaf tissue of 115 regenerated plants, was 47%. Resistance tests and Southern analysis of seed progenies of three diploid transgenic plants demonstrated homozygous integration of multiple copies of the transgene at one locus at least in the first plant, heterozygous integration at one locus in the second plant and heterozygous integration at two loci in the third plant.Abbreviations PEG polyethylene glycol - T₀ regenerated transgenic plant - GUS -glucuronidase - CaMV cauliflower mosaic virus - ARE anaerobic responsive element - OCS octopine synthase - T₁ first generation progeny of transgenic plants     

Electrostatic surface properties of plasmalemma vesicles from oat and wheat roots. Ion binding and screening investigated by 9-aminoacridine fluorescence

Right-side-out and sealed plasmalemma vesicles were isolated from roots of spring wheat (Triticum aestivum L. cv. Drabant) and oat (Avena sativa L. cv. Brighton) by two-phase partition in a medium containing sucrose (0.25 mol l^-1). Oat root plasmalemma vesicles were discovered to contain a strongly fluorescent compound with an emission maximum at 418 nm. The surface potential of the membranes was monitored by 9-aminoacridine fluorescence and the effect of protein concentration, mannitol versus sucrose, absence of osmoticum, concentrations of salt, and titrations with chelators investigated. It is concluded that i) protein concentrations of less than 50 g ml^-1 for oat and 100 g ml^-1 for wheat plasmalemma vesicles should be used to avoid serious problems with non-linearity of response of 9-aminoacridine fluorescence, ii) mannitol can be used instead of sucrose as the osmoticum, iii) the vesicles were ruptured in the absence of osmoticum allowing us to monitor both sides of the membranes, iv) plasmalemma vesicles from oat roots are more negative than vesicles from wheat roots, and v) oat and wheat root plasmalemma vesicles are isolated with about the same amounts of bound Ca²⁺ and Mg²⁺. These bound divalent cations may not, however, reflect the in-vivo conditions since the tissues were homogenised in the presence of ethylenediaminetetraacetic acid.Abbreviations EDTA ethylenediaminetetraacetic acid - EGTA ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid - c1/2 value concentration at which half of the maximum effect is observed - Mops 3-(N-morpholino)propanesulfonic acid     

Die campaniformen Sensillen im Pedicellus der Florfliege (Chrysopa,Planipennia)

Zusammenfassung Der Pedicellus der Florfliege (Chrysopa) enthält an seinem Distalende fünf campaniforme Sensillen. Sie bestehen aus vier Zellen: einer Sinneszelle, einer trichogenen Zelle (= accessory supporting cell Stuart u. Satir, 1968) und zwei Hüllzellen. Im rezeptorischen Fortsatz der Sinneszelle wurzelt ein umgewandeltes Cilium, dessen Distalende von einer cuticularen Scheide umhüllt wird.

---

On the campaniform sensilla of the pedicel of Chrysopa

Summary At the apex of the pedicel in Chrysopa there are five campaniform sensilla, which are arranged in a cycle. They are composed of four cells: two enveloping cells, a trichogen cell (= accessory supporting cell, Stuart and Satir, 1968) and one sense cell. The distal nerve process contains a transformed cilium. The tip of the cilium with the so called tubular body is enveloped by a cuticular sheath.

---

---

Die Untersuchung wurde mit Unterstützung durch die Deutsche Forschungsgemeinschaft durchgeführt.     

Monitoring the effect of subunit assembly on the structural flexibility of human alpha apohemoglobin by steady-state fluorescence

A single energy transfer distance, between the sole intrinsic tryptophanyl donor [14 (A12)] and a nonfluorescent sulfhydryl acceptor probe (4-phenylazophenylmaleimide, PAPM) attached to the only cysteine [104 (G11)], has been employed to examine the effect of subunit assembly on the structure of the heme-free human-hemoglobin. Efficiencies of energy transfer were measured in 0.05 M potassium phosphate buffer,pH 7.0, at 5°C, and the structural flexibility of-apohemoglobin, in the absence and presence of human-heme-containing chains, was examined by a steady-state solute quenching technique. The quenched efficiencies (E ^O) and Förster distances (R ₀ ^O ) were analyzed by least-squares to determine the goodness of fit ( _R ² ) for the assumed distribution parameters: average distance ¯r and half-widthhw. Data for-apohemoglobin in the absence and presence of ^h chains yielded values for ¯r of 18 and 22 Å andhw of 20 and 8.5 Å, respectively. Although the increase in ¯r for-apohemoglobin in the presence of ^h chains was presumably a consequence of additional quenching from the heme moiety, the change in the half-width strongly indicated a decrease in the flexibility of the-apohemoglobin chain within the assembled protein. A transition in structural flexibility similar to that demonstrated here may be an important aspect of human hemoglobin assembly.     

Ionophore-mediated transmembrane movement of divalent cations in small unilamellar liposomes: An evaluation of the chlortetracycline fluorescence technique and correlations with black lipid membrane studies

Summary Conceptual advances in the field of membrane transport have, in the main, utilized artificial membranes, both planar and vesicular. Systems of biological interest,viz., cells and organelles, resemble vesicles in size and geometry. Methods are, therefore, required to extend the results obtained with planar membranes to liposome systems. In this report we present an analysis of a fluorescence technique, using the divalent cation probe chlortetracycline, in small, unilamellar vesicles, for the study of divalent cation fluxes. An ion carrier (X537 A) and a pore former (alamethicin) have been studied. The rate of rise of fluorescence signal and the transmembrane ion gradient have been related to transmembrane current and potential, respectively. A second power dependence of ion conduction-including the electrically silent portion thereof — on X537 A concentration, has been observed. An exponential dependence of current on transmembrane potential in the case of alamethicin is also confirmed. Possible errors in the technique are discussed.