Observations on the micro-anatomy of crystallolithus hyalinus gaarder and markali

Summary The fine structure of Crystallolithus hyalinus (Gaarder and Markali), known to be the motile stage of Coccolithus pelagicus (Wallich) Schiller (Parke and Adams 1960), has been investigated for purposes of comparison with Chrysochromulina and Prymnesium, recently recognised as probably related to it. Anoptral contrast light microscopy of living cells and electron microscopy of thin sections and shadow-cast whole mounts show that there is a close resemblance to species of Chrysochromulina not only in cell structure but also in the details of haptonema structure and scale structure. The origin of the scales from internal vesicles has been traced as in other genera but the holococcoliths (crystalloliths) do not appear to arise in this way. The mode of origin of the crystalloliths is thus unknown, but it is suggested that they may be developed outside the cell from calcite secreted in solution into the appropriate cavity. Their constant position between a layer of scales and an outer enveloping skin is consistent with this and one unfamiliar cytoplasmic organelle, composed of peculiar thick-walled tubes within some large superficial vesicles, is described as perhaps involved in this process. Apart from these special features the resemblance to both Chrysochromulina and Prymnesium is sufficient to add considerable weight to Christensen's suggestion (Christensen 1962) that a new group should be defined to accommodate these genera.     

Ultrastructural study on the morphogenesis of the neuromuscular junction in the skeletal muscle of the chick

Summary The morphogenesis of the neuromuscular junction was examined at the ultrastructural level in the skeletal muscle of the lower limb of the chick. The fine structure of the neuromuscular junction of the adult fowl was essentially the same as that in other vertebrates; the junction consists of the axon terminal, the Schwann cell, and the muscle fiber. The first visible sign of neuromuscular junction formation, in embryos of 13 days in ovo, was the membrane thickening of the sarcolemma which develops into the postsynaptic membrane. The axons approaching the muscle fibers were incompletely ensheathed by a Schwann cell and contained vesicles. The subsequent differentiation of the junctional sarcoplasm, the axoplasm, and the Schwann cell cytoplasm takes place from 13 to 18 days in ovo and the junction nearly reaches maturity at around 20 days in ovo. The formation of complicated anastomoses and branching of the junctional infoldings seems to occur after hatching. These ultrastructural observations are in good agreement with histochemical findings (cholinesterase method) in terms of the chronology of the morphogenesis of the junction.This investigation was supported in part by U. S. Public Health Service Grant MH 12269-01, administered by Dr. Kazuo Ogawa. It was initiated on the suggestion of Prof. J. Nakai, Department of Anatomy, Faculty of Medicine, Tokyo University, and a part of it was performed in his laboratory. The author is greatly indebted to Prof. K. Ogawa, Department of Anatomy, Kansai Medical School, for his guidance and encouragement, and to Dr. S. Igarashi, Department of Anatomy, Tokyo University, for some technical advice.     

The fine structure of micrococcus radiodurans

Summary The fine structure of the radiation-resistant bacterium, Micrococcus radiodurans, isolated by Anderson, was studied by electron microscopy of intact and disrupted cells using thin sectioning and negative staining techniques. The cytoplasm and nuclear structures are normal, but the cell wall and sheath are more complex than any so far described for a bacterium. The surface consists of four distinct layers, each having a characteristic fine structure, one of which has been tentatively identified as that responsible for maintaining the rigidity of the cells. Striations with a periodicity of 175 to 200 Å are visible in thin sections of this layer, and a pseudo-hexagonal array of dark holes is seen in surface view of negatively-stained fragments. It is concluded that this layer is the main structural element of the cell wall of M. radiodurans. The other three layers of the surface have not been clearly located in thin sections; one of these layers has a well-defined pattern of hexagonally arranged units similar to that observed in Spirillum serpens by Murray but with different dimensions.Sir Halley Stewart Research Fellow.     

Biochemical and structural studies of a l-haloacid dehalogenase from the thermophilic archaeon Sulfolobus tokodaii

Haloacid dehalogenases have potential applications in the pharmaceutical and fine chemical industry as well as in the remediation of contaminated land. The l-2-haloacid dehalogenase from the thermophilic archaeon Sulfolobus tokodaii has been cloned and over-expressed in Escherichia coli and successfully purified to homogeneity. Here we report the structure of the recombinant dehalogenase solved by molecular replacement in two different crystal forms. The enzyme is a homodimer with each monomer being composed of a core-domain of a β-sheet bundle surrounded by α-helices and an α-helical sub-domain. This fold is similar to previously solved mesophilic l-haloacid dehalogenase structures. The monoclinic crystal form contains a putative inhibitor l-lactate in the active site. The enzyme displays haloacid dehalogenase activity towards carboxylic acids with the halide attached at the C2 position with the highest activity towards chloropropionic acid. The enzyme is thermostable with maximum activity at 60°C and a half-life of over 1 h at 70°C. The enzyme is relatively stable to solvents with 25% activity lost when incubated for 1 h in 20% v/v DMSO.     

Fine structure of antheridia and oogonia of Phytophthora macrospora,the downy mildew fungus of rice plants

In the present paper fine structure of antheridium and oogonium ofPhytophthora macrospora (Sacc.)S. Ito etI. Tanaka, the downy mildew fungus of rice plants was discussed.Before the fertilization some nuclei and a large number of mitochondria were scattered in the cytoplasm of the antheridium. Many lipid granules were observed in the peripheral region, but vacuoles did not appear at this stage of antheridium. Many mitochondria were associated in the neighborhood of the fertilization pore. The wall at the pore was very thin, but the wall surrounding the pore was slightly swollen towards the inside.In the oogonium, many nuclei, mitochondria and cytoplasmic matrix were observed at the peripheral part. A large number of lipid granules was found in the oogonium, but they were more numerous in the peripheral region. The vacuoles developed as the oogonium matured. They were enveloped by tonoplast and contained vacuolar matrix. Many electron dense granules were in contact with the tonoplast or free in the vacuoles, and they were larger in the central part. As stated above, wall at the fertilization pore was thin. However, the oogonial wall surrounding the pore swelled protruding into the oogonium. An electron-dense layer was recognized between the antheridial and oogonial wall, and the walls of both the organs were closely in contact with each other.Contribution No. 252.     

Ultrastructural study of the regressing prothoracic glands of blattarian insects

Summary The prothoracic glands, source of the molting hormone ecdysone, regress within a few days after the final molt, a process which was analyzed with electron microscopic methods in the cockroaches Leucophaea and Blaberus. This strictly timed event is accompanied by drastic alterations in cellular fine structure. Early signs of breakdown appear in groups of nuclei whose substance becomes segregated into patches of contrasting electron density characteristic of pyknosis.The most conspicuous change in the cytoplasm of parenchymal cells concerns the appearance of large, heterogeneous inclusion bodies in which various cellular elements become segregated. These compartments seem to represent autophagic vacuoles within which the gradual degradation of much of their contents takes place, presumably under the influence of lysosomal enzymes. Undigested swirls of membranous character may remain sequestered within these packets for some time.At advanced stages of cellular atrophy, plasma membranes and nuclear envelopes have gradually disappeared, and masses of protoplasm undergoing autolysis become invaded by a greater number of hemocytes than are present in nymphal glands. These phagocytic elements appear to engulf debris of parenchymal cells as well as some degenerating connective tissue elements. After the completion of the regressive process, the axial band of musculature characteristic of the nymphal gland persists on its own. Whether or not some parenchymal cells (or possibly their precursors) capable of reactivation persist in the proximity of this muscle is unknown.The resorption of the prothoracic gland in the newly emerged insect is the result of physiological autolysis and seems to be aided by the activity of phagocytic hemocytes.Dedicated to Professor W. Bargmann on his 60th birthday in friendship and admiration.This study was supported by Research Grants AM-03984, NB-02145 and NB-05219 from the U.S.P.H.S.I wish to express my thanks to Mrs. S. Wurzelmann, Mrs. C. Jones, Mrs. C. Grubman, and Mr. S. Brown for their excellent technical assistance.     

Jimpy, a new totally sex-linked gene in the house mouse

Summary A new recessive mutant jimpy, jp, is described. It is situated on the differential segment of the sex chromosome and is lethal in the hemizygous condition. The jimpy mice show a behaviour disorder distinguished by an intention tremor first visible at about 11 days; after about three weeks they are liable to go into a convulsion if disturbed; death occurs usually between twenty-five and thirty-two days. jp shows about 21% recombination with Ta and 36% with Bn. It has not been tested directly against Br and Mo, but they both have been shown to be about four units beyond Ta with respect to Bn (Falconer 1954). The presumed order is therefore Bn, Ta, Br and Mo, jp.Member of the Scientific Staff, Medical Research Council.     

On the infiltration of Golgi bodies from the follicular epithelium to the egg

Summary 1. In a well advanced oocyte of the tortoise the egg membranes besides the theca and the single-layered epithelium consist of a zona pellucida often differentiated into zona striata and a homogeneous layer; underlying these two layers is a layer of cortical fibrillae or fibrillar layer, Next to this layer, is the limiting membrane of the egg which is not present in all stages and generally disappears in a well developed oocyte. In certain animals either the homogeneous layer or fibrillar layer is absent. 2. In certain animals,Golgi bodies seem to be extruded into the follicle cells from the theca cells. 3. At a particular stage of development the follicle cells become very active and produce a large number of smallGolgi bodies. TheseGolgi granules filter through canalicular passages of the zona radiata into either the homogeneous layer and from thence into the fibrillar layer or where a homogeneous layer is not present directly to the fibrillar layer. Where a fibrillar layer is not present they are transferred directly to the limiting membrane and from thence to the egg. 4. In certain cases e. g. in Fowl, Calotes and Uromastix, fairly large lumps ofGolgi bodies are extruded from the follicle cells through the zona pellucida into the egg. Here the fine canilicular passages do not seem to form a vehicle for the passage of these comparatively larger bodies. 5. The fine canalicular passages in the zona radiata ofTestudo graeca andKachuga smithii and the fibrillar prolongation of the cytoplasm which we have called the fibrillar layer are marked features of the egg membranes at certain stages of development of the egg. During the period when infiltration ofGolgi bodies through these passages takes place slides prepared by silver nitrate and osmic methods show black beaded chains ofGolgi granules in various stages of descent. 6. It is claimed that the extrusion and infiltration ofGolgi bodies from the follicular epithelium to the egg are established phenomena at least in the Vertebrates.     

Fine structure of the vagina,accessory glands,uterus, oviducts and Gene's organ in the unfed tick,Ornithodoros (Pavlovskyella) erraticus (Ixodoidea: Argasidae)

The fine structure of the reproductive system of the unfed femaleOrnithodoros (Pavlovskyella) erraticus is described. The vagina consisting of vestibular (vv) and cervical (cv) regions is formed of a single epithelial layer lined with cuticle and surrounded by muscle layers. Epithelial cells ofvv show no signs of activity, while those ofcv have structural features of transporting epithelia. A pair of tubular accessory glands opening at the junction of the two vaginal regions consist of a layer of microvillate columnar cells containing rough endoplasmic reticulum; this is possibly involved in the formation of colloidal material which presumably coats the egg surface during its passage through the vagina. The bilobed uterus opening into the cervical vagina secretes material that is possibly responsible for the release of sperm from the endospermatophore. The paired oviducts joining the uterine lobes anteriorly have distinctive distal (do) and proximal (po) parts as well as ampullae (amp) at their junction.do cells contain many lysosome-like structures which probably play a role in breaking down some sperms ascending to the ovary. Cells ofamp are packed with rickettsia-likeWolbachia, while those ofpo contain a few granules. Gene's organ, present in female ticks only, consists of a stalk and two horns of epithelium lined internally by a cuticular layer. Bundles of microtubules are detected in the horn cell cytoplasm. This organ functions to waterproof the eggs with a waxy layer just before they are deposited.     

The surface of the leaf in normal and glossy maize seedlings

Summary The wettability of leaf surface in maize seedlings may vary according to the genotypes,Gl orgl. Techniques in electron microscopy have made it possible to resolve the fine structure of theGl—surface as contrasted with those ofgl ₁,gl ₂,gl ₃, andgl ^H . The normal surface, shows minute projections which are almost absent in the glossy surface of young seedlingsgl ₁; thegl ₂,gl ₃ andgl ^H seedlings present a somewhat intermediate situation.With 1 Figure in the Text     

Ultrastructure of globule leucocyte inclusions in the rat and mouse

Summary The fine structure of the globules in globule leucocytes of rat and mouse infected with various parasites has been studied. Best preservation was obtained with glutaraldehyde and subsequent osmium fixation. It is concluded that the structure of the globules is host specific. In rat, the structure is not parasite specific, but in mouse slight differences in the structure of the globules of the globule leucocyte in different experimental conditions suggest a small change in the packing of the sub-units.The authors are grateful to Professor G. M. Wyburn and Mr. R. N. C. Aitken for research facilities and advice. The work was partially financed by Agricultural Research Council Grant No. AG 17/49.     

Differential uptake of fumarate by Candida utilis and Schizosaccharomyces pombe

The dicarboxylic acid fumarate is an important intermediate in cellular processes and also serves as a precursor for the commercial production of fine chemicals such as l-malate. Yeast species differ remarkably in their ability to degrade extracellular dicarboxylic acids and to utilise them as their only source of carbon. In this study we have shown that the yeast Candida utilis effectively degraded extracellular fumarate and l-malate, but glucose or other assimilable carbon sources repressed the transport and degradation of these dicarboxylic acids. The transport of both dicarboxylic acids was shown to be strongly inducible by either fumarate or l-malate while kinetic studies suggest that the two dicarboxylic acids are transported by the same transporter protein. In contrast, Schizosaccharomyces pombe effectively degraded extracellular l-malate, but not fumarate, in the presence of glucose or other assimilable carbon sources. The Sch. pombe malate transporter was unable to transport fumarate, although fumarate inhibited the uptake of l-malate. Received: 15 March 2000 / Received revision: 4 July 2000 / Accepted: 9 July 2000     

A polysaccharide fraction from the red seaweed Champia novae-zealandiae Rhodymeniales,Rhodophyta

The polysaccharide recovered after extraction of Champia novae-zealandiae is a galactan with alternating 3-linked d-galactopyranosyl units sulfated at the 2-position, and 4-linked galactopyranosyl units sulfated at both the 2- and 3-positions that are predominantly of the l- and partly of the d-configuration. Other minor substitution includes 6-O-methyl ether or 4,6-pyruvate acetal on the 3-linked residues. Techniques used in determining the structure include infrared and ¹³C-NMR spectroscopy, and GC-MS analysis of alditol acetate derivatives produced by reductive hydrolysis/acetylation of native, methylated, and/or desulfated samples. These results are of particular interest because 4-linked 2,3-desulfated galactosyl residues have not been encountered as major constituents of red algal polysaccharides.     

β-Carbon stereoselectivity of N-carbamoyl-d-α-amino acid amidohydrolase for α,β-diastereomeric amino acids

N-Carbamoyl-d-α-amino acid amidohydrolase (d-carbamoylase) was found to distinguish stereochemistry not only at the α-carbon but also at the β-carbon of N-carbamoyl-d-α-amino acids. The enzyme selectively acted on one of the four stereoisomers of N-carbamoyl-α,β-diastereomeric amino acids. This simultaneous recognition of two chiral centers by d-carbamoylase was useful for the fine stereoselective synthesis of α,β-diastereomeric amino acids such as threonine, isoleucine, 3,4-methylenedioxyphenylserine and β-methylphenylalanine. The stereoselectivity for the β-carbon was influenced by the pH of the reaction mixture and by the bulk of the substituent at the β-carbon. Received: 18 June 1999 / Received revision: 30 July 1999 / Accepted: 6 August 1999     

Submicroscopic structure of the membranous labyrinth

Summary Investigations were performed by light and electron microscope on the submicroscopic structure of the epithelium of Corti's organ in the white rat.Morphological and structural differences between the inner hair cells and the outer hair cells are revealed.The inner hair cells are closely inter-related with the inner supporting cells and have a polyhedral shape, whereas the outer hair cells look like cylinders and are surrounded by an intraepithelial fluid.The structural peculiarities consist of differences in the dimensions of the hairs, in the arrangement of cytoplasmic organoids and in the aspect of the receptoneural junction. In both sensory hair cells 4 zones of different structure can be distinguished from the surface inwards: apical zone, intermediate zone, perinuclear zone and receptoneuronal junction. The functional value of these different zones is discussed and compared with what has been demonstrated in other receptors.The pillar cells and the Deiters' cells are supporting cells which have a filamentous skeleton, composed of submicroscopic individual filaments. These filaments have a diameter of about 215 Å and present some analogies with the tonofilaments of the stratified squamous epithelium. The filaments are arranged differently in the pillar cells and in the Deiters' cells. Possible functional differences between these patterns are discussed.The reticular membrane is not an extracellular cuticle. It consists of intracellular cementitious material (like the terminal bars of the epithelial cells).The Hensen's and Claudius cells, the Böttcher's cells, the inner supporting cells, the inner and outer spiral sulcus cells are regular prismatic cells with few endoplasmic organoids and without filaments.This work is dedicated to the memory of the late Prof. L. Pietrantoni.     

Die Feinstruktur der Cerebraldrüse von Planorbarius corneus L. (Basommatophora)

Summary The fine structure of the cerebral gland of the fresh water snail Planorbarius corneus has been described. The apical cell surfaces of the epithelium bordering the lumen are covered with short microvilli and in some parts with cilia. There is a remarkable enlargement of the basal surface of those cells underlying the perineurium by long branched cell processes. The fine structure and density of the substances in the lumen differ; inclusions of paracrystallin structure are found. The possible functional significance of these organs in Pulmonates is discussed.

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Frau Prof. Dr. B. Scharrer zum 60. Geburtstag gewidmet.

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Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

The wound response in the siphonous algaCaulerpa simpliciuscula C. Ag.: II. The effect of wounding on carbon flow

Summary Following on from a previous study on changes in cytology and fine structure during the wound response in the siphonous green algaCaulerpa simpliciuscula (Dreher, Grant, andWetherbee 1978), changes in the carbon metabolism during this wound response have been studied. There was a decrease in the rate of photosynthesis and an increase in the rate of respiration immediately on wounding, but rates of both photosynthesis and respiration returned to those of unwounded tissues within 6 hours. Wounding depressed the rate of starch synthesis and sucrose synthesis but increased the rate of synthesis of soluble 1,3 ß-glucan, lipid and sulphated polysaccharide. When the flow of carbon from these various compounds was studied by means of pulse chase experiments, it was found that only sucrose and sulphated polysaccharide showed different kinetics in control and wounded tissue. The changes which were observed are consistent with the direct involvement of sulphated polysaccharides in the formation of structures formed during the wound healing process.     

Gloeomonas oderChlamydomonas? Elektronenmikroskopische Untersuchungen anGloeomonas simulans

The fine structure ofGloeomonas simulans Fott (1957) was studied electron microscopically to ascertain whether it belongs to the genusChlamydomonas rather than toGloeomonas. Most cytoplasmic elements and the cell wall do not differ from otherChlamydomonadaceae but its flagellar rootlet system is unique: Each of the two flagella has an accessory basal body; its basis is accompanied by two inner and two outer bands which are connected distally (one inner and one outer band on each side) resp. proximally (the two outer bands); the latter form a long (up to 3–5 µm) connecting band between the two flagellar bases. The nucleus contains fibrillar bundels.—The unique flagellar rootlet system seems to provide a better basis for the generic classification ofGloeomonas than the position of the contractile vacuoles or the size of the apical papilla, and strongly suggests the exclusion ofG. simulans fromChlamydomonas.

Zweiter Beitrag einer vonEttl (1965a) begonnenen Publikationsreihe.     

The induction of flowering in vitro in stem segments of Plumbago indica L.

Summary Internode segments excised from vegetative Plumbago indica plants are responsive to photoperiodic treatments in vitro. Under long days, they produce vegetative buds; under short days, they develop inflorescences. These inflorescences can remain devoid of flowers (vegetative inflorescences), or produce normal flowers which open in the test tubes. The minimum duration of the short-day treatment capable of inducing flowering is of the order of 4 weeks.The production of inflorescences under short days is affected by various factors. An adequate level of sucrose is necessary. Sucrose can be replaced by maltose and, to a small extent, by cellobiose, but not by mannitol or lactose. Auxins and gibberellins inhibit the production of flower buds, whereas cytokinins and adenine do not. Guanine, thymine, cytosine or uracil alone are ineffective, but thymine or its precursor, orotic acid, enhance the production of floral buds when adenine and kinetin are also present in the medium. Several amino acids, as well as glutamine and asparagine, tend to reduce inflorescence formation at 3×10^-4 M or above; urea increases it slightly at the same concentrations. Both the cis- and the trans-isomer of abscisin II enhance inflorescence formation under short days, but have no such effect under long days.High concentrations of adenine re-established the red coloration of the petals which is typical of the clone used. Otherwise, the color of the flowers grown in vitro was pink, presumably because of the depressing effect of kinetin on anthocyanin synthesis.The techniques used have been described in the preceding article (Nitsch and Nitsch, 1966). All segments were excised from internodes of stock plants kept in completely vegetative conditions in a greenhouse by means of 16-hour photoperiods.