Characterization of proteins from Ascaris lumbricoides which bind specifically to carboxypeptidase.

Inhibitors of the peptidase and esterase activities of carboxypeptidases A and B have been isolated from extracts of Ascaris lumbricoides var suis. These proteins were obtained by treatment of the aqueous extracts at low pH, precipitation with ammonium sulfate, molecular sieving on Bio-Gel P-4, and chromatography on DEAE-cellulose. The inhibitors were resolved into three homogeneous peaks on CM-cellulose. These components, CM-A, CM-B, and CM-C, have constant specific activity and were recovered in a 41% yield. They moved as single bands when subjected to electrophoresis at high or low pH on polyacrylamide gels and they have similar amino acid compositions. Methionine, tyrosine, and cysteine are absent from each of the inhibitors. The 65 residues of CM-B suggest a minimum molecular weight of 7530, in close agreement to the value of 7600 +/- 200 determined on a Bio-Gel P-100 column. Each of the proteins has the same NH2-terminal residues, NH2-Asx-Glx-Val-Glx- and the same COOH-terminal residue, leucine. A plot of per cent acrylamide versus log relative mobility suggests that the three proteins are charge isomers. CM-B appears to be stable to high NaCl concentrations, extremes of pH, high temperatures, and digestion by intestinal proteases. Carboxypeptidase C, carboxypeptidase N, and yeast protease C are not inhibited by CM-B. However, the exopeptidase and esterase activities of human carboxypeptidase A are inhibited. The inhibitors appear to bind to bovine carboxypeptidase A with an atypical stoichiometry. Two moles of CM-B inhibitor bind to 1 mol of enzyme. The evidence is: (a) a demonstrated purity of bovine carboxypeptidase A, (b) minimal and maximal inhibitor molecular weights by different methods, of 7600 and 8300, and (c) a maximum specific activity of apparently homogeneous inhibitors which is 50% of that predicted for unit stoichiometry.     

Cytophotometric study of nuclear differentiation during pollen development in Tradescantia Paludosa

Summary During pollen development the dry weight, total protein, histone, DNA, arginine, and lysine content were analysed by cytophotometric methods in partially isolated nuclei. The amount of analysed substances increased from the end of the meiosis to the mitosis of the microspores to the double of the initial values. After mitosis the ratio histone/DNA remained almost unchanged in both vegetative and generative nuclei. On the other hand a large difference in the ratio non-histone protein/DNA could be observed, the vegetative nucleus containing more non-histone protein than the generative nucleus. The rate of RNA synthesis being higher in the vegetative nuclei, these non-histone proteins may have some function in nuclear activation. The DNA of the generative nucleus is duplicated before anthesis, whilst in the vegetative nucleus the DNA content remains constant.     

Porphyrins in the perienteric fluid of Ascaris lumbricoides.

Cain G. D. and Bassow F. 1976. Porphyrins in the perienteric fluid of Ascaris lumbricoides. International Journal for Parasitology6: 79–82. Porphyrins in the perienteric fluid of adult female A. lumbricoides were esterified in methanolic H₂SO₄, extracted in chloroform, separated by thin-layer chromatography, and identified spectrophotometrically before and after conversion to their zinc and copper chelates. Protoporphyrin IX was the major component, comprising 95·4% of the total; the remaining 4·6 % was coproporphyrin III. Uroporphyrin was not detected; no porphyrins were recovered from other worm tissues. Fluid from worms with light and dark colored guts varied in protoporphyrin content from 0·58 to 4·08 nmoles/ml, respectively, but fluid from both groups contained similar molar ratios of protoporphyrin, coproporphyrin and heme.     

Spermatogenesis and spermiogenesis in Ascaris lumbricoides Var. suum

Reorganization of the prophase I nucleus marks the beginning of the first meiotic division. A pair of centrioles is present at each pole at metaphase I and mitochondria are not observed in the spindle area. A chromosomal pellicle, which resembles a kinetochore plate but has no apparent association with microtubules, surrounds each autosome at metaphase I and II. The sex body lags behind the autosomes at anaphase I and segregates differentially to one daughter cell. Mitochondria and a pair of centrioles are present in the spindle during the second meiotic division. Localized condensation of chromatin and fusion of the condensed chromatin of the secondary spermatocyte telophase nucleus results in a compact spermatid nucleus. Loss of spermatid cytoplasm is effected by the ejection of a cytophore vesicle.     

Developmentally programmed nuclear destruction during yeast gametogenesis

Autophagy controls cellular catabolism in diverse eukaryotes and modulates programmed cell death in plants and animals. While studies of the unicellular yeast Saccharomyces cerevisiae have provided fundamental insights into the mechanisms of autophagy, the roles of cell death pathways in yeast are less well understood. Here, we describe widespread developmentally programmed nuclear destruction (PND) events that occur during yeast gametogenesis. PND is executed through apoptotic-like DNA fragmentation in coordination with an unusual form of autophagy that is most similar to mammalian lysosomal membrane permeabilization and mega-autophagy, a form of plant autophagic cell death. Undomesticated strains execute gametogenic PND broadly in maturing colonies to the apparent benefit of sibling cells, confirming its prominence during the yeast life cycle. Our results reveal that diverse cell-death-related processes converge during gametogenesis in a microbe distantly related to plants or animals, highlighting gametogenesis as a process during which programmed cell death mechanisms may have evolved.     

Funktionsmorphologie des Exkretionsorgans des Spulwurms Ascaris lumbricoides L.

Zusammenfassung Das Seitenkanalsystem von Ascaris lumbricoides wurde elektronenmikroskopisch untersucht. Beim erwachsenen Tier erstreckt sich das in den lateralen Epidermisleisten eingebettete einzellige Organ vom Nervenring bis etwa zur Körpermitte. Im 2. Körperviertel besitzt es kein durchgehendes Kanallumen und erscheint degeneriert. In allen übrigen Bereichen (mit Ausnahme des Ausführungskanals) besitzt es den gleichen Aufbau aus zwei Schichten. Die das Kanallumen begrenzende innere Schicht enthält zahlreiche extraplasmatische Räume, von denen zumindest die am weitesten innen liegenden mit dem Kanallumen kommunizieren. Die äußere Zellmembran besitzt viele Einfaltungen, von denen einige weit in das Cytoplasma hineinragen. Der Gewebeanteil der lateralen Epidermisleisten, der dem Seitenkanalsystem unmittelbar anliegt, enthält sehr viele Interzellularräume, die ein zusammenhängendes Drainage-System bilden. Zur histochemischen Lokalisation von ATP-ase-Aktivität wurden Experimente durchgeführt. Die möglichen Mechanismen der Bildung der Exkretflüssigkeit werden diskutiert unter Berücksichtigung bereits veröffentlichter physiologischer Befunde.

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Ultrastructure and function of the excretory organ of Ascaris lumbricoides L. (Nematoda)

Summary The Excretory organ (H-system) of Ascaris lumbricoides has been investigated electronmicroscopically. In adult animals this single-cell-organ embedded in the lateral lines extends from the nerve ring to approximately the middle of the body.In the second quarter of the body it lacks a continuous canal lumen, and it seems to be degenerated. In all of the other regions (except the stem leading to the excretory pore) it consists of two zones. The inner zone lining the canal lumen contains several extraplasmatic spaces; at least those placed the farthest inside communicate with the canal lumen. The outer cell membrane shows many infoldings, some of which extend deeply into the cytoplasm. The tissue of the lateral line adjacent to the canal system contains very many intercellular spaces which build a coherent intracellular rainage-system. Experiments have been performed in order to localize the ATPase activity histochemically. Possible mechanisms for the forming of the excretory fluid are discussed under consideration of physiological results already published.

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Abkürzungen Ak Ausführungskanal - Bm Basalmembran - Cp Cytoplasmaplatten - lE linke Epidermisleiste - rE rechte Epidermisleiste - Ef Einfaltungen der äußeren Zellmembran - Fb Fibrillenbündel - Go Go Golgiapparat - Hg Hüllgewebe - Is lamelläre Interzellularsubstanz - Iz Interzellularraum - Kl Kanallumen - K Kutikula - Lh Leibeshöhle - Mu Muskelzelle - Mi Mitochondrien - Ms mittlerer Gewebestreifen (= Mittelstreifen) der Epidermisleiste - Mt Mikrotubuli - N Zellkern - No Nucleolus - Ne Nervenring - eP elektronendichte Partikel - sP sphärische Partikel - P Kernpore - Q Querbalken - epR extraplasmatischer Raum - Lho Längsholm - Mf Membranfusion - äS äußere Schicht des Seitenkanalsystems - iS innere Schicht der Längsholme - Sy syncytiale Cytoplasmamasse ohne Interzellularen - V Verzweigungskanal - iZ innere Zone um einen Verzweigungskanal Inauguraldissertation der Mathematisch-Naturwissenschaftlichen Fakultät der Freien Universität Berlin (gekürzt). Herrn Prof. Dr. G. Kümmel danke ich für die Anregung zu diesem Thema und für sein ständiges kritisches Interesse der Untersuchung, Frau C. S. Friedemann für die Anfertigung der Zeichnungen und Fräulein H. Schmidt für technische Assistenz.     

Ultrastructural analysis of sex determination in Ascaris lumbricoides var. suum

At metaphase of the first spermatocyte meiotic division in Ascaris lumbricoides var. suum the polymorphic bivalents are arranged at the equatorial plate peripheral to a central aggregate mass. The aggregate mass segregates to only one of the two daughter cells and thus behaves as a sex chromosome. The mass is comprised of two components: a granular chromatin that can be differentiated from the autosomes at metaphase I, and a condensed chromatin that has a similar morphology to the autosomes but is functionally differentiated at anaphase I. The condensed chromatin may be autosomes that have fused with the sex chromsomes and may be responsible for the segregation of the whole aggregate at anaphase I. Analysis of the origin of the mass is discussed and microtubular association in the movement of the mass is described.