Ultrastructural autoradiographic study of the intracellular fixation of3H-acrolein

Summary The fixation of³H acrolein inDunaliella bioculata was followed by means of quantitative autoradiography electron microscopy. A fixation was observed mostly in the nucleus where aldehyde linkage remained stable at least for 48 hours.     

Einfluss einiger lipophiler Lösungsmittel in gasförmigem Zustand auf die CO2-fixierung durch Luzerne

Summary Vapourized organic solvents such as: diethyl ether, methylene chloride, carbon disulfide, benzene, and n-octane have an inhibiting effect on CO₂ fixation in alfalfa leaves (Medicago sativa). One exception is CS₂ which stimulates overall fixation up to 10 vol%. Beyond 10 vol% it inhibits overall CO₂ fixation like the other solvents, but stimulates the formation ofl-alanine as the major remaining fixation product. Proportionality exists between water insolubility of the organic solvents and their inhibiting effect on CO₂ fixation.     

Laser microbeam-induced fixation for electronmicroscopy: Visualization of transient developmental features in nematode embryos

Summary In order to study development of embryos ofCaenorhabditis elegans at an ultrastructural level, a new method of fixation has been developed. With a laser microbeam coupled to a microscope the impermeable eggshell is punctured to allow penetration of the fixative. At specific stages of embryogenesis further development can be arrested at will under visual control. As fixation occurs instantaneously, transient events (e.g. different phases of mitosis and cytokinesis) can be visualized.     

Influence of phosphorus supply and light intensity on mycorrhizal response inPisum-Rhizobium-Glomus symbiosis

The influence of mycorrhizal colonization withGlomus mosseae on parameters of N₂ fixation and plant growth was studied in pot experiments with pea plants (Pisum sativum L.) infected withRhizobium leguminosarum and supplied with varied levels of phosphorus (P) and nitrogen (N). Reduced light intensities were used to evaluate the dependence of the microsymbionts on assimilate supply. In plants grown with low P supply, mycorrhization increased the concentration of P in shoots, and thus N₂ fixation. Reduced light intensity significantly depressed mycorrhizal colonization and nodule growth in low-P plants. When P supply did not limit plant growth and N₂ fixation, however, the percentage of mycorrhizal colonization was reduced due to the higher P status, and the microsymbionts were not impaired by low light intensities. To maximize carbohydrate supply, another experiment was carried out at high light intensity of 900 mol m^–2s^–1 and with non-limiting P supply. Nitrogen fertilization, given as starter N, enhanced plant growth, but delayed nodule formation. Towards flowering, nodulation rapidly increased, but less so inGlomus inoculated plants. After 28 days mycorrhizal plants were lower in shoot dry weight, nodule dry weight and nitrogenase activity. The results suggest that under many, but not all, environmental conditions the host plant is able to restrict mycorrhizal colonization and, thus, to prevent impairment ofRhizobium symbiosis.deceased in May 1994     

Sequential analysis of Giemsa banded chromosomes inVicia faba

Summary Vicia faba (broad bean) root-tip chromosomes were subjected to the SSC-Giemsa and trypsin-Giemsa regimes. Phase-lucent cross-bands seen after fixation/SSC treatment subsequently stain positively with Giemsa. Sequential analysis of the trypsin-Giemsa regime shows, in contrast, that bands are manifest through selective removal of material from interband regions.     

Raster-elektronenmikroskopische Untersuchungen an der Hausstaub- und Asthma-MilbeDermatophagoides pteronyssinus (Trouessart, 1897) (Acarina: Astigmata)

Summary Various methods of fixation ofDermatophagoides pteronyssinus for the REM were tested. OsO₄ vapor yielded the best results. The morphology of male and female mites is described, based on REM exposures.

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Naturhistorisches Museum Basel.     

Avantages et inconvénients de la fixation par perfusion du tissu nerveux à la lumière d'une donnée de cytochimie

Summary It has been extensively demonstrated that perfusion is the best fixation procedure for providing good morphological evidence ofstructures, especially in the case of nervous tissue. But it might be questioned if it should also be preferred whencytochemical data have to be obtained or compared.As a preliminary attempt to answer this question, lipoproteic inclusions in nerve cells of vegetative ganglia of adult cat have been considered. These occur in about 25% of cells after immersion in fixative, whereas if perfusion is used, a relation appears between the amount of saline perfused before the fixative fluid and the percentage of cells containing inclusions: this falls to 10% after 300 ml saline, to 1% after 800 ml. In conclusion, though structural artefacts have been avoided and fixation appears excellent, perfusion is significantly responsible for a definite cytochemical alteration.     

Ultrastruktur der Ovozyten vonOrnithodorus moubata,Murray (Ixodoidea:Argasidae) nach Triple-Fixation

Summary The triple-fixation (simultaneous fixation in a mixture of glutaraldehyde and osmium tetroxide and post-treatment in aqueous uranylacetate) is especially suitable for the demonstration of membranes and to distinguish different types of vesicles in the oocytes of the soft tickOrnithodorus moubata.

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Die Arbeit wurde vom Schweiz. Nationalfonds unterstützt.

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Frau S.Latscha-Stoller danke ich für ihre unentbehrliche technische Mitarbeit.     

De l'emploi d'isotopes radioactifs artificiels,dans le but d'exercer un effet radio-biologique localisé

Summary One of the authors has previously reported on a method which consists in the utilization of an artifical radioactive isotope (Zn⁶³), suspended in a suitably prepared solution ofpectin, for the production oflocalized biological radiation effects.This « macromolecular occlusion » of the radioactive isotope enables one to perform intraperitoneal injections (in cases of cancer of the ovaries with severe metastatic peritoneal extension), evidently also instillations in cavernous organs, and furthermore direct intratumoral injections, without diffusion of the radioactivity outside the treated areas, as shown both by autoradiographs and controls of blood and urine specimens with a Geiger counter.The authors investigated further whether this procedure would also be suitable for obtaining, by means ofintravenous injections, alocalized radiation effect within thelungs, as presumably the radiozinc, held in the large molecules of pectin, could thus be retained in the pulmonary capillaries. Intravenous injections of such a pectin solution containing radiozinc were performed on rabbits, and autoradiographic controls gave evidence of this expected fixation within the lungs.For the purpose of preliminary clinical investigation 40 millicuries of Zn⁶³ suspended in 6 cm³ of a 3 p. c. isotonic pectin solution were injectedintravenously in a female patient with mainly pulmonary metastases of a previously operated hypernephroma. This patient had been also submitted to X-ray therapy. In spite of a poor general condition, the injection was well tolerated. Autoradiographic controls showed quite clearly that the radioactivity remains precisely localized within the pulmonary areas. No radioactivity whatsoever was demonstrated with the counter in the urine eliminated by this patient after the injection, a fact which points to a rather amazing accuracy of the fixation of the radiozinc in the lungs. This first clinical experience seems quite interesting in view of improving the therapeutic possibilities of pathological, especially neoplastic pulmonary conditions.     

Does ammonium uptake influence xylem sap composition inPhaseolus vulgaris L. andGlycine max (L.) Merill?

The question of whether ammonium uptake influences the occurrence of ureides in legumes has been addressed in this study by investigating threeP. vulgaris genotypes as well as one cultivar ofGlycine max. All plants were raised in sand culture during the dry season in northern Thailand and irrigated daily with nitrogen-free nutrient solution, or the same solution containing 12 mol m^–3 nitrogen in the form of (NH₄)₂SO₄ or KNO₃, each treatment consisting of different proportions of either compound. Regression analyses of xylem sap composition relative to ammonium vs. nitrate supply of plants harvested at V4, R1 and R6 indicated close positive correlations of xylem amino nitrogen content and negative correlations with xylem nitrate content and ammonium supply. Statistically significant correlations between relative xylem ureide content and ammonium availability could be established for theP. vulgaris cultivar Brilliant up to stage R1, but not for the other plants investigated. It was concluded that at least for some genotypes of common bean a relationship exists between ureide production and ammonium uptake by the root system. Since the extent to which ureide production is stimulated remains quite small, its relevance to the xylem solute technique for measurement of N₂ fixation may be limited. Nevertheless, due to the possibility of large genotypic differences in the impact of ammonium on ureide production, this factor must be considered in calculations if N₂ fixation is to be determined in soils containing significant amounts of ammonium, e.g. in paddy fields.     

The C4-dicarboxylate transport system ofRhizobium meliloti and its role in nitrogen fixation during symbiosis with alfalfa (Medicago sativa)

TheRhizobium meliloti C₄-dicarboxylate transport (Dct) system is essential for an effective symbiosis with alfalfa plants. C₄-dicarboxylates are the major carbon source taken up by bacteroids. Genetic analysis of Dct^– mutant strains led to the isolation of thedct carrier genedctA and the regulatory genesdctB anddctD. The carrier genedctA is regulated in free-living cells by the alternative sigma factor RpoN and the two-component regulatory system DctB/D. In addition, DctA is involved in its own regulation, possibly by interacting with DctB. In bacteroids, besides the DctB/DctD system an additional symbiotic activator is thought to be involved indctA expression. Further regulation ofdctA in the free-living state is reflected by diauxic growth of rhizobia, with succinate being the preferred carbon source. The tight coupling of C₄-dicarboxylate transport and nitrogen fixation is revealed by a reduced level of C₄-dicarboxylate transport in nitrogenase negative bacteroids.     

High genetic variability despite haplodiploidy in primitive sawflies of the genusCephalcia (Hymenoptera,Pamphiliidae)

Hymenoptera are haplodiploid and usually display very low genetic variation. Most data concern social or parasiticApocrita, while the little information available for the primitive phytophagous species of the suborder Symphyta is contradictory. The present study is related to seven species of the genusCephalcia, living in coniferous forests of Northern Eurasia and sharing spruce (Picea sp. pl.) as host plant. Individuals from 22 populations belonging toCephalcia abietis, C. alashanica, C. arvensis, C. erythrogaster, C. fallenii, C. fulva, C. klugii from Europe and China were surveyed for genetic variation at 28 loci using enzyme electrophoresis. Pairs of sibling species were recognized withinC. arvensis andC. fallenii, corresponding to different phenological and morphological forms. In the latter case, reproductive isolation in sympatry occurs despite low genetic distance (D=0.059). Large genetic distances and fixed alternate alleles were observed between Chinese and European populations ofC. abietis andC. arvensis. Expected heterozygosity ofCephalcia populations (0.197, SD 0.064) is significantly higher than that of other Symphyta (Tenthredinoidea) (averageH _exp 0.059, SD 0.032) (two-tailed Mann-Whitney test, Z=4.39, p<0.01). These data suggest that haplodiploidy per se does not reduce the genetic variation in mostCephalcia populations. Most of the factors that can lower the potential for genetic diversity in a haplodiploid genetic system are not so effective inCephalcia populations, which seem to be comparable to diplodiploid insect populations in diversity. In a few isolated populations the large number of fixed loci and the large genetic distances may support the predicted faster rate of fixation, as a consequence of haplodiploidy.     

A quantitative method for measuring the uptake of colloidal carbon by mouse tissues

Résumé On décrit une méthode spectrophotométrique quantitative pour la détermination de la répartition du carbone colloÏdal dans les organes d'une seule souris. La sensibilité de la méthode est de 0,25l de la suspension de carbone colloÏdal C11/1431a. La rate est le tissu qui a présenté la plus grande capacité de fixation du carbone. Elle est suivie par la moelle osseuse, la foie et les poumons.     

Reinterpretation of the ultrastructure of cartilage matrix.

The epiphyseal cartilage from new-born mouse was treated with collagenase in two ways: either before fixation or after glutaraldehyde fixation. The electron dense granules of the matrix were not seen in the micrographs of cartilage treated with collagenase before fixation. It is concluded that collagen plays a definite role in the formation of the granules at the time of tissue fixation and that the granules are fixation artifacts.     

Colorants acides et détermination embryonnaire chez les Echinodermes

Summary The effects of three dyes with carboxyl groups are been studied on the developing eggs of the sea urchin,Paracentrotus lividus. Uranin and rose Bengal are both some derivative of the xanthene. Chrome violet CG is a carboxyl derivative of pararosolic acid. In dilute solutions these three dyes induce the development of radial larvae. At higher concentration, uranin and rose Bengal are very effective animalizing agents. Rose Bengal is more active than uranin. Rose Bengal differs from uranin by the fixation of halogens on the molecule of dye which increase the acidity of the carboxyl group. It appears that animalizing activity runs parallel with acidity. The results of these experiments are discussed in relation to the animalizing effects of various acid sulfonated dyes. The possible significance of the combinations between acid dyes and basic components of the cells in the animalization is considered.     

Reinterpretation of the ultrastructure of cartilage matrix

Summary The epiphyseal cartilage from new-born mouse was treated with collagenase in two ways: either before fixation or after glutaraldehyde fixation. The electron dense granules of the matrix were not seen in the micrographs of cartilage treated with collagenase before fixation. It is concluded that collagen plays a definite role in the formation of the granules at the time of tissue fixation and that the granules are fixation artifacts.Commonwealth Academic Staff Fellow. Permanent address: Department of Zoology, University of Poona, Poona 411007, India.     

Zur Gestalt des Influenza-Virus

Summary Influenza-Virus particles are generally considered to be of spherical shape after conventional fixation in OsO₄. However, after fixation with KMnO₄, the elementary bodies of the Influenza-Virus (Type A Singapur I/57) — adsorbed on the surface of erythrocytes — exhibit hexagonal shape in ultrathin-sections.     

Ultrastructural autoradiographic study of the intracellular fixation of 3H-acrolein.

The fixation of 3H acrolein in Dunaliella bioculata was followed by means of quantitative autoradiography electron microscopy. A fixation was observed mostly in the nucleus where aldehyde linkage remained stable at least for 48 hours.     

Comparative morphometric study of the cells of the third proximal segment of the rat kidney under different conditions of fixation.

The morphometric study of the P3 segment of rat proximal tubule has shown that neither the method of fixation, nor the fixative itself, significantly change the relative and absolute volumes of the cell compartments, provided that the fixation media are made approximately isotonic.     

Significance of ‘empty vesicles’ in postganglionic sympathetic nerve terminals

Résumé Nous avons montré que dans les terminaisons nerveuses sympathiques de l'iris de chat, la proportion de vésicules à contenu osmiophile (noradrénaline) dépend essentiellement de la technique de fixation. Par ailleurs, après incubation des iris dans des solutions de noradrénaline et après fixation convenable, la quasi-totalité de ces vésicules renferment de la noradrénaline. Nous concluons que ces vésicules représentent une population potentiellement homogène, toutes étant capables de stoquer la noradrénaline.