Fibrous Vesicles Research Articles

Supramolecular Assembly of Hybrid Pt(II) Porphyrin/Tomatine Analogues with Different Nanostructures and Cytotoxic Activities.

A simple strategy for synthesizing supramolecular hybrids was developed for the preparation of bioavailable nanohybrid photosensitizers by assembling visible-light-sensitive Pt(II) meso-tetrakis(4-carboxyphenyl)porphyrinporphyrin (PtTCPP)/tomatine analogues. The hybrids were self-assembled into nanofibrous or nanosheet structures approximately 3–5 nm thick and several micrometers wide. α-Tomatine generated a unique fibrous vesicle nanostructure based on intermolecular interactions, while dehydrotomatine generated nanosheet structures. Nanoassembly of these fibrous vesicles and sheets directly affected the properties of the light-responsive photosensitizer for tumor photodynamic therapy (PDT), depending on the nanostructure of the hybrid PtTCPP/tomatine analogues. The cytotoxicity of PtTCPP to cancer cells under photoirradiation was significantly enhanced by a tomatine assembly with a fibrous vesicle nanostructure, attributable to increased incorporation of the drug into cells.

An ultrastructural study of carpospores in the red alga Chondrus pinnulatus (Harvey) okamura, 1930 (Rhodophyta: Gigartinaceae) from Vostok Bay, the Sea of Japan

The morphological features of carpospores in the red alga Chondrus pinnulatus have been studied using methods of transmission and scanning electron microscopy. Rounded mature carpospores are assembled into groups. Each carpospore is surrounded by a two-layered mucilage wall. The electron-dense cytoplasm contains numerous starch grains, fibrous vesicles, and large clusters of fibrous vesicles. The plastids have well-developed thylakoids and the cell nucleus occupies a nearly central position. The nucleolus is large and loose and is localized near the nuclear membrane. Dictyosomes, small fibrous vesicles, osmiophilic granules, and plastids are localized at the periphery. Mitochondria are arranged near the dictyosomes, plastids, and around the nucleus. A generalized scheme of the fine structure of the carpospore has been proposed for red algae on the basis of our own and literature data.

Morphology and Phylogeny of Prorocentrum texanum sp. nov. (Dinophyceae): A New Toxic Dinoflagellate from the Gulf of Mexico Coastal Waters Exhibiting Two Distinct Morphologies.

A new planktonic species of Prorocentrum is described from the Gulf of Mexico. First observed with the Imaging FlowCytobot, Prorocentrum texanum sp. nov. was characterized using LM, SEM, and TEM along with sequencing of the SSU, LSU, and ITS ribosomal regions and the mitochondrial cob and cox1 regions. P. texanum sp. nov. is a round to oval bivalvate dinoflagellate, with a prominent anterior, serrated solid flange on periflagellar a platelet and an opposing short, flat flange on the h platelet. The periflagellar area consists of 10 platelets. Both left and right valves have shallow round depressions and two-sized valve pores. The anterior ejectosome pore pattern differs between the left and right valve in relation to the periflagellar area and margins. Ten to eleven rows of tangential ejectosome pores are present on each valve. P. texanum sp. nov. has two varieties which exhibit distinct morphotypes, one round to oval (var. texanum) and the other pointed (var. cuspidatum). P. texanum var. cuspidatum is morphologically similar to P. micans in surface markings, but is smaller, and has a serrated periflagellar flange, and is genetically distinct from P. micans. Cytologically, P. texanum has two parietal chlo-roplasts, each with a compound, interlamellar pyrenoid, trichocysts, fibrous vesicles that resemble mucocysts, pusules, V- to U-shaped posterior nucleus, golgi, and tubular mitochondria. No genetic difference was found between the two varieties in the five genes examined. Phylogenetic analysis of the SSU, LSU, and ITS ribosomal regions place P. texanum sp. nov. as a sister group to P. micans. One isolate of P. texanum var. texanum produces okadaic acid.

New taxa of the Porphyridiophyceae (Rhodophyta): Timspurckia oligopyrenoides gen. et sp. nov. and Erythrolobus madagascarensis sp. nov

Yang E.C., Scott J. West J.A., Orlova E., Gauthier, D., Küpper F.C., Yoon H.S. and Ulf Karsten U. 2010. New taxa of the Porphyridiophyceae (Rhodophyta): Timspurckia oligopyrenoides gen. et sp. nov. and Erythrolobus madagascarensis sp. nov. Phycologia 49: 604–616. DOI: 10.2216/09-105.1Two new marine unicellular red algae are described: Timspurckia oligopyrenoides gen. et sp. nov. isolated from southeastern Australia and Erythrolobus madagascarensis sp. nov. isolated from Madagascar. Timspurckia oligopyrenoides cells are spherical, 7–11 µm in diameter, greyish red to reddish brown and surrounded by a conspicuous fibrillar matrix about 2 µm thick. Cells exhibit positive phototaxis. In the single chloroplast the lobes extend from several pyrenoids to occupy most of the cell. A peripheral thylakoid is absent. The pyrenoid matrices are filled with tubular thylakoids and are usually surrounded by starch sheaths in the adjacent cytoplasm. The nucleus is peripheral with a nucleolus appressed to the nuclear envelope usually closest to the cell center. Golgi bodies are associated with mitochondria usually at the cell periphery. Erythrolobus madagascarensis cells are spherical, 5–8 µm in diameter, greyish red to reddish brown and surrounded by a thin conspicuous fibrillar sheath. Cells exhibit positive phototaxis. Each cell has a single chloroplast with several lobes extending from an eccentric pyrenoid forming a complex parietal layer. A peripheral thylakoid is absent. The pyrenoid matrix is filled with tubular thylakoids, and a starch sheath is visible in the adjacent cytoplasm. The nucleus is eccentric and has a central nucleolus. Golgi bodies are associated with mitochondria usually at the cell periphery. Numerous conspicuous, electron-transparent, fibrous vesicles are always present in the cells. The sequence divergences of psaA and psbA genes and phylogeny support that E. madagascarensis sp. nov. and T. oligopyrenoides gen. et sp. nov. are newly uncovered natural entities within the class Porphyridiophyceae along with Erythrolobus coxiae, Porphyridium spp. and Flintiella sanguinaria. The low-molecular-weight carbohydrates (LMWCs) floridoside, digeneaside and trehalose are present in both taxa. Thus, three independent features – ultrastructural characters, plastid gene sequences and LMWC patterns – support the suggested phylogeny and taxonomy of these red algae.

Ultrastructure of single cells, callus-like and monospore-like cells inPorphyra yezoensis ueda on semisolid culture medium

It had been demonstrated that individual cells or protoplasts isolated fromPorphyra thallus by enzyme could develop into normal leafy thalli in the same way as monospores, and that isolated cells develop in different way in liquid and on semi-solid media. The authors observed the ultrastructure of isolated vegetative cells cultured on semi-solid media and compared them with those of monospores and isolated cells cultured in liquid media. The results showed that subcellular structures were quite different among cells in different conditions. In their development, isolated cells on semi-solid media did not show the characteristic subcellular feature of monospore formation, such as production of fibrous vesicles. Callus-like cells formed on semi-solid media underwent a distinctive modification in cellular organization. They developed characteristic cell inclusions and a special 2-layer cell covering. Golgi bodies, ER, starch grains, mitochondria. Vacuoles were not commonly found in them.

Differentiation of Male Gametes in Gracilaria verrucosa (Rhodophyta, Gracilariales)

The development of male gametes (spermacia) in the red alga Gracilaria verrucosa has been studied using methods of transmission electron microscopy. Early spermatangia located along the wall of the conceptacle show an elongated shape in the thin sections. In the central part of the electron-dense cytoplasm of these cells there is a nucleus; numerous fibrous vesicles are arranged in the periphery. During the process of differentiation, the spermatangia become more rounded in shape and a large spermatangial vesicle is developed. The subsequent development of spermatium is accompanied by polarization of the spermatangium and the subsequent excretion of the spermatangial vesicle. The spermatia are oval cells containing a nucleus and fibrous vesicles. The process of differentiation of male gametes in G. verrucosa does not differ from that in five species of the genus Gracilaria, where it has already been studied. However, any conclusions about the degree of similarity between the spermatia in all the studied species can be made only after a detailed comparative analysis of the ultrastructural characteristics of these gametes.

The ultrastructure of separated and cultured cell ofPorphyra yezoensis

There are many reports that cells (protoplasts) separated from the thallus ofPorphyra by enzyme can develop to normal leafy thalli in the same way as monospores. But there are few investigations on the subcellular structure of the isolated vegetative cell for comparison with the subcellular structure of monospores. To clarify whether the separated and cultured cells undergo the same or similar ultrastructure changes during culture and germination as monospores undergo in their formation and germination, we observed their ultrastructure, compared them with those of the monospore and found that the ultrastructure of separated and cultured cells did not have the characteristic feature as that of monospore formation, such as production of small and large fibrous vesicles, but was accompanied by vacuolation and starch mobilization like that in monospore germination. The paper also discusses the relations between monospores and separated and cultured cells.

Evidence of sexual reproduction inSmithora naiadum(Erythropeltidales, Rhodophyta) and its evolutionary significance

Smithora naiadum blades, which grow epiphytically on seagrasses, are monoecious gametophytes. Previous reports of male gametes are confirmed using light and electron microscopy. Spermatogenesis is initiated when vegetative cells of the monostromatic thallus divide anticlinally twice to give an associated group of four cells. Each of these cells then divides periclinally producing a distromatic thallus. A subsequent, unequal periclinal division of each of these cells forms a small cell that becomes the spermatangium and a larger cell that becomes the carpogonium. Abundant fibrous vesicles are produced during spermatogenesis. Female gametes and fertilization are reported for the first time. The fate of the zygote remains unresolved, but available evidence suggests germination into a diminutive, as yet unknown, life history stage. Because of its systematic placement, near the root of the red algal phylogenetic tree, the occurrence of sexual reproduction in Smithora has significant evolutionary implications.

Fine structure of initial calcification during odontogenesis in the dog salmon (Oncorhynchus keta, Teleostei).

Histological examinations of tooth germs in juvenile dog salmon (chum salmon), Oncorhynchus keta, were carried out by transmission electron micoscopy in order to observe fine structures of initial calcification sites in dental matrix and to determine the relationship between electron-dense fibrous substances and matrix vesicles reported by Igarashi (1983). A number of matrix vesicles round or oval in shape and 300-700 A in diameter appeared in the apical dental matrix around the odontoblasts during the stage of matrix formation and about half of them contained more electron-dense crystal-like structures. On the other hand, matrix vesicles relatively decreased in number, but electron-dense fibrous substances consisting of flocculent substances and fine granular substances became well-developed and longer and increased in number in the apical dental matrix distant from the odontoblasts. These fibrous substances also contained more electrondense crystal-like structures. The stereophotographs showed that the fibrous substances exhibited filmy structures in three dimensions and continued to more electron-dense crystal-like structures. Moreover, the electron-dense fine granular substances would seem to attach to these filmy structures in parts. It seems that these filmy structures are the initial form of crystal. A number of matrix vesicles existed near the fibrous substances and some came into contact with the fibrous substances. A state indicative of the unit membrane of these matrix vesicles having broken and part of the contents disappeared was often obtained. It seems that these fibrous substances appeared later than matrix vesicles because of the observation in the early stage of matrix formation. These observations lead us to the conclusion that the fibrous substances derived from the contents of matrix vesicles. Fibrous substances indicated a positive reaction with ruthenium red staining. No fibrous substances appeared in the basal dental matrix during the stage of matrix formation in the basal portion in spite of the appearance of a number of matrix vesicles and small aggregates of crystal-like structures. It seems that this difference is closely concerned with the fact that the apical dental matrix is the initial portion to begin the calcification of tooth.

Ultrastructural study of Pisdnoodinium pillulare (Schäperclaus, 1954) Lom, 1981 with special emphasis on its attachment to the fish host

Abstract. Trophonts of Pisdnoodinium pillulare (Schäperclaus, 1954), a common ectoparasite of freshwater aquarium fish, are attached to host cells by means of a specialized structure, the attachment disc. Unlike other dinoflagellate genera parasitic on fish and invertebrates, this disc features nail‐like organelles, the rhizocysts. Head‐parts of the rhizocysts are inverted in separate compartments, rhizothecas, in the sole of the disc while their long shafts are firmly embedded in the cytoplasm of cells of the host epidermis or gill epithelium. The attachment inflicts a serious injury on the host cells which may ultimately be destroyed. Rhizocysts originate in the subnuclear cytoplasm from where they migrate into the attachment disc. There are other specialized organelles and inclusions; fibrous vesicles, membraneous bodies, striated tubular bodies and paracrystalline bodies. Pisdnoodinium has well‐developed chloroplasts. While its cytological adaptations indicate a nutritional dependence on the host, there is no evidence of ingestion of host‐derived particulate material. Pisdnoodinium may derive an essential part of its nutrition from photosynthesis.

Fine structure of swimming, encysting, and germinating spores of Haliphthoros milfordensis

Electron microscope observations were made of swimming, encysting, and germinating zoospores of two strains of Haliphthorus milfordensis Vishniac isolated from gills of the white shrimp, Penaeus setiferus, and from the American lobster, Homarus americanus. Although structurally similar to zoospores of other Oomycetes, those of H. milfordensis have certain distinguishing features including: mitochondria grouped anteriorly around the nucleus, the presence of electron-dense vesicles which become transformed into unique "scalloped-fringed" vesicles, and an anterior arrangement of most cell organelles with the posterior of the zoospore being filled with numerous fibrous vesicles. During spore encystment, flagellar retraction and rounding up of the spore is accompanied by a central positioning of most cell organelles (central nucleus surrounded by concentric bands of rough endoplasmic reticulum), and ends with formation of a unique, spined, trilaminate wall. Cyst germination occurs via fracturing of the spined wall, with repeated emergence of zoospores from the spore cysts noted in addition to the more normal germ tube formation.

A comparative account of cyst coat ontogeny in saprophytic and fish-lesion (pathogenic) isolates of the Saprolegnia diclina – parasitica complex

The ultrastructural development of encystment vesicles (formerly bar bodies) and the outer cyst coat has been examined in primary and secondary spores of a range of isolates of Saprolegnia, mostly from the S. diclina – parasitica complex, of both saprophytic and fish-lesion origin. The development of both primary encystment vesicles (PEVs) in zoosporangia and secondary encystment vesicles (SEVs) in primary cysts appears essentially similar. The vesicles are probably derived from the rough endoplasmic reticulum system. The tubular primary spines and solid secondary boathooks seem to form autonomously within the vesicle matrix. Upon settling all of the preformed encystment vesicles are liberated from the zoospores to give rise to an electron-dense, amorphous, outer cyst coat, in which the spines are embedded. Fish-lesion isolates of S. parasitica have longer tubules on their primary cysts than saprophytic isolates and have distinctive bundles of long (<10 μm) boathooks on their secondary cysts compared with the single short (< 1.0 μm) boathooks typical of the saprophytes. However, even in fish-lesion isolates considerable variation in SEV and boathook sizes was observed. Staining by the Thiéry procedure revealed a high concentration of polysaccharidelike material with both the encystment vesicle cortex layer and spines, although treatment with various glucanases and proteases failed to digest either component. Spherical fibrous vesicles also appear in primary cysts and their contents are partially removed by protease digestion. These appear morphologically similar to the glycoprotein adhesive vesicles described in other oomycetes and appear to be discharged by secondary zoospores. The complex encystment apparatus in Saprolegnia is compared with that described in other oomycete and chytridiomycete fungi and the possible phylogenetic significance discussed. The role of the long boathook spines in relation to the infection of fish is also considered.

The Ultrastructure of Carposporogenesis in Gigartina teedii (Roth) Lamour. (Gigartinales, Rhodophyceae): Gonimoblast Cells and Carpospores

Carpospore differentiation has been studied in Gigartina teedii using light and electron microscopy and histochemistry. Young gonimoblast cells possess a rich and dense protoplasmic content, but contain few organelles. The chloroplasts are present in a proplastid state and usually possess only a single peripheral thylakoid. Mature gonimoblast cells are highly vacuolate. Carpospore development may be separated into 3 stages. The youngest (stage I) is characterized by dictyosomes with a straight profile, formation of fibrous vesicles, and the presence of mucilage sacs, dense ER-associated bodies, microbodies and twisted structures within the mitochondrial matrix. Formation of the internal thylakoid system commences in the chloroplasts, while concomitantly the first small-sized starch grains appear in the cytoplasm. The primary mucilage of mucilage sacs is rich in sulfated polysaccharides and probably serves in spore liberation. The second stage (stage II) is characterized by the disappearance of the mucilage sacs, dense ER-associated bodies, microbodies and twisted structures of mitochondria. The dictyosomes retain a straight profile, exhibit extreme hypertrophy and continue to secrete fibrous vesicles. The inflated fibrous vesicles derived from dictyosomes of stage I and II migrate to the periphery of the cytoplasm and are responsible for most of the cell wall deposition to form the thick carpospore wall. During the final stage (stage III) the dictyosomes assume a semicircular profile and produce cored vesicles with a glycoproteinrich content, which probably functions as an adhesive after spore liberation. The older carpospores are also characterized by having large quantities of starch, rough ER, and numerous large fibrillar vacuoles which are derived from the Golgi-cored vesicles. The mucilaginous component of the cell wall and the intercellular matrix of Gigartina teedii carpospores seem to be rich in sulfated polysaccharides.

ULTRASTRUCTURE CHARACTERISTICS OF MONOSPORE FORMATION IN PORPHYRA GARDNERI (RHODOPHYTA)1

ABSTRACTMonosporogenesis in Porphyra gardneri (Smith & Hollenberg) Hawkes has been examined with the transmission electron microscope. Increased dictyosome activity resulting in the production of small and large fibrous vesicles is a characteristic feature of the developing monospores. Released monospores lack a cell wall. Four of the six Porphyra species commercially cultivated in Japan form monospores. Monospore production is of economic importance because it results in an increased number of foliose thalli.

Cuticle formation in parasitic nematodes: Ultrastructure of molting and the effects of actinomycin-D

The ultrastructure of cuticle formation and the effect of actinomycin-D on cuticle formation were examined in the second molt of Nippostrongylus brasiliensis . At the end of the first molt the rough endoplasmic reticulum of the hypodermis (epidermis) had narrow cisternae. As cuticle formation began the RER cisternae dilated and accumulated a fibrous material. These large fibrous material-containing vesicles then decreased and by the time cuticle deposition was completed they had disappeared. Actinomycin-D prevented the proliferation of RER and the formation of fibrous material-containing vesicles. Actinomycin-D induced several ultrastructural alterations in the hypodermis, the most prominent of which were large crystalline-like inclusions. Other alterations involved the configuration of RER and structure of nucleoli. These morphological aspects of cuticle formation were correlated to the molecular events that control molting.

THE ULTRASTRUCTURE OF CARPOSPOROGENESIS IN THE MARINE HEMIPARASITIC RED ALGAERYTHROCYSTIS SACCATA1

SUMMARYThe ultrastructure of carposporogenesis forErythrocystis saccatais described. The fusion and gonimoblast cells contain few organelles, and chloroplasts are in a proplastid state, with pit plugs between gonimoblast cells dissolving early in development. Carpospore development may be separated into 3 stages, the first stage being characterized by the appearance of straight‐profiled dictyosomes, fibrous vesicles, and an increase of discoid thylakoids within the chloroplasts. During the second, stage the dictyosomes assume a curved profile and striped vesicles are formed by the endoplasmic reticulum. The third stage is initiated by the disappearance of striped vesicles and the appearance of straight‐profiled dictyosomes secreting vesicles with cores. Mature carpospores consist of many cored vesicles, fibrous vesicles, and floridean starch grains. A single wall layer surrounds each carpospore since the carposporangial wall becomes incorporated into a mucilaginous matrix surrounding the spores.

Golgi vesicles of uncommon morphology and wall formation in the red alga, Polysiphonia.

Golgi bodies of immature carposporangia ofPolysiphonia sp. are composed of a polarized stack of six to ten curved cisternae. The cisternae are surrounded by 50–200 nm diameter slightly granular vesicles. Hypertrophied, fibrillar Golgi cisternae occur in mature carposporangia. Secretory vesicles originate from ends of cisternae and by complete vesiculation of terminal cisternae; 0.6–1.2 μm diameter, fibrous vesicles, many with electron dense nucleoids are abundant throughout the cytoplasm of mature sporangia. Vesicles expand, fuse with each other and cluster around starch granules. Some vesicles secrete their content into the spore wall. Morphological analyses of starch granules as well as topographical relations between vesicles, starch granules and the adjacent cytoplasm suggest that these Golgi vesicles function like lysosomes. The significance of these observations is discussed in relation to the composition of plant cell walls and cellular expansion.

The ultrastructure of carpospore differentiation in the parasitic red alga Levringiella gardneri (Setch.) Kylin

Three cytological stages can be recognized during carpospore differentiation in Levringiella gardneri. The youngest stage (stage 1) is identified by a single nucleus surrounded by numerous mitochondria, initiation of dictyosome activity, fibrous vesicles formation, and chloroplasts in the peripheral cytoplasm. During stage 2 striated vesicles are formed within the endoplasmic reticulum, dictyosomes are semicircular in profile and vesicles secreted by dictyosomes contribute either to wall deposition or fibrous vesicle formation. In stage 3 carpospores, striated vesicles degenerate, and straight-profiled dictyosomes secrete numerous cored vesicles. Fibrous vesicles, which are formed throughout carpospore differentiation, release their contents to the outside of the cell and contribute to wall deposition. During the latter part of stage three, however, microtubules appear and these are oriented at right angles to the plasmalemma. A mature carpospore is surrounded by a carpospore wall and a carposporangial wall. One nucleus, an abundance of floridean starch, cored vesicles, mitochondria, chloroplasts and some fibrous vesicles are found in the cytoplasm. Dictyosome activity is low in mature carpospores. Mucilaginous material surrounds all carposporangia and this material is secreted by dictyosomes in secretory cells, located beneath the per icarp cells.

ULTRASTRUCTURE OF TETRASPOROGENESIS IN THE PARASITIC RED ALGA LEVRINGIELLA GARDNERI (SETCHELL) KYLIN12

SUMMARYUltrastructural studies on tetraspore formation in Levringiella gardneri revealed that 3 stages may be recognized during their formation. The youngest stage consists of a uninucleate tetraspore mother cell with synaptonemal complexes present during early prophase of meiosis I. Mitochondria are aggregated around the nucleus, dictyosome activity is low, and chloroplasts occur in the peripheral cytoplasm. A 4‐nucleate tetraspore mother cell is formed prior to tetrahedral cell cleavage, and an increase in the number of chloroplasts and mitochondria occurs. Small straight‐profiled dictyosomes secrete vesicles into larger fibrous vesicles or contribute material to the developing tetraspore wall. During the second stage of tetraspore formation, striated vesicles form within endoplasmic reticulum, semicircular profiled dictyosomes secrete vesicles for fibrous vesicles or wall material, and starch formation increases. The final stage is characterized by the disappearance of striated vesicles, presence of straight, large dictyosomes which secrete cored vesicles, and an abundance of starch grains. Cleavage is usually complete at this stage and the tetraspore wall consists of a narrow outer layer of fibrillar material and an inner, electron transparent layer. These spores are surrounded by a tetrasporangial wall which was the original wall surrounding the tetraspore mother cell.

CHRONIC SEMINAL VESICULITIS AND PROSTATITIS

In reporting a series of cases of about 200 patients operated on in association with Dr. F. W. Smith in 1923, the symptomatology of seminal vesicle disease was classified into three groups with reference to symptoms. The first group included those cases in which pain is the predominating element with inflammatory changes; the second, those cases in which rheumatic symptoms are present, and the third, those cases in which vague local symptoms are pronounced with an accompanying unexplained neurosis. Three years' further experience bears out the value of this grouping as a working scheme. Operative indications may be similarly epitomized, and these three clear and distinct reasons for surgical intervention and drainage may be given: (1) the evacuation of pus, (2) the relief of pain and (3) the removal of hard, indurated fibrous vesicles productive of systemic, prostatic and bladder symptoms. GROUP I: PAIN AND INFLAMMATORY CHANGES Infection of the