Ultrastructure of transitional epithelium of man

Summary Blocks of human normal renal pelvis and ureter obtained at the time of surgery were fixed in glutaraldehyde and osmium with or without ruthenium red, for electron microscopic observations. The transitional epithelium is arranged in three cell layers: basal, intermediate and superficial. All epithelial cells show numerous microvilli and contain the characteristic vesicles of transitional epithelium, bundles of cytoplasmic filaments, microtubules and numerous free ribosomes. The epithelial extracellular compartment is notably large and appears as an intricate, tridimensional network of canaliculi and cisternae which are wider in the intermediate and superficial layers and in which microvilli and cytoplasmic folds of vicinal cells are often attached or interdigitated. At these sites there are desmosomes.The surface of all transitional epithelial cells is covered by a fibrillar mucous coat which is more developed at the plasmalemma of the free border of luminal cells in which microvilli are also seen. Ruthenium red stains selectively the plasmalemma and the mucous coat of the free surface of the epithelium, indicating the presence of an acid polysaccharide. With this technic (Luft, 1965), it is observed, radiating from the plasmalemma, branching filaments which measure 100 Å in diameter forming a zone of varying density which is about 400 m wide and which corresponds, at the light microscopic level, to the luminal border of the transitional epithelial cells in which a sialomucin has been identified. The slender filaments have a beaded appearance. At the free border, superficial cells are attached by functional complexes in which tight junctions seal the epithelial intercellular space, which is opened at the level of the basement membrane where only desmosomes are observed.The ultrastructure of human transitional epithelium of urinary tract resembles the duct cells of the salt gland of certain marine birds (Fawcett, 1962) and the amphibian epidermis (Farquhar and Palade, 1965) in which there are active processes of transport. The mucous surface coat, selectively stained by the ruthenium red, contains a sialomucin (Monis and Dorfman, 1965, 1967).The ultrastructure and histochemistry of the mucous fluffy coat of man transitional epithelium and the observations of Porter and Tamm (1955), on the ultrastructure of preparations of the Tamm and Horsfall mucoprotein (1952) are bases for suggesting that transitional epithelium of urinary tract of man is the site of biosynthesis of certain urinary mucoids. Present investigations are directed to obtain evidence to substantiate this hypothesis.General Abbreviations B basal cell - E exfoliating cell - I intermediate cell - L lumen - S superficial cell - SC surface coat - bm basement membrane - ci cell infolding - d desmosome (macula adhaerens) - f fibroblast - fi cytoplasmic filaments - is intercellular space - jc junctional complex - ly lysosome - lym lymphocyte - mt microtubules - m mitochondria - mv microvilli - n nucleus - r ribosomes - rv round vesicle - zo zonula occludens - za zonula adherens Dr. Monis wishes to thank Dr. E. De Robertis for the use of the electron microscope facilities of the Instituto de Anatomía General y Embriologia, Facultad de Medicina, Universidad de Buenos Aires. — Prof. E. Trabucco and Dr. R. J. Borzone (Cátedra de Clinica Genitourinaria de la Facultad de Medicina, Universidad de Buenos Aires) generously supplied the specimens which were the bases of this study. — Thanks are due to Mrs. A. M. Novara and Mrs. Defilippi-Novoa for efficient technical help and to Miss Rosa Gentile for secretarial assistance. Photomicrography by Mr. M. A. Saenz.Dr. Zambrano is investigator (CNICT).     

Breeding and post-breeding structure of the vas deferens of the long-toed salamander Ambystoma macrodactylum

The vas deferens of Ambystoma macrodactylum is composed of a peritoneal epithelium, connective tissue layer with fibroblasts, circular smooth muscle, capillaries, cells containing lipid, and a luminal epithelium composed of a single layer of cuboidal cells covered by a net of interconnected ciliated squamous cells. The cuboidal cells have abundant rough endoplasmic reticulum, mitochondria, and PAS + secretory vesicles. Squamous cells of breeding males consistently have tufts of ～100 cilia located at one end of the long axis of each cell. These cilia may help distribute secretory products. The squamous cells, absent in post-breeding males, are apparently sloughed into the lumen. Lipid vesicles are present throughout the cytoplasm of the cuboidal and squamous epithelial cells and are also in some cells of the connective tissue layer. These vesicles increase dramatically in number during the first 4 weeks after breeding and may serve as an energy pool for the next breeding season. Enzyme-histochemical tests for testosterone synthesis were negative. In addition to the accumulation of lipid and the loss of squamous cells in the vas deferens, after breeding PAS + vesicle production is terminated. These alterations appear to represent energy conservation strategies employed by the sperm-depleted vas deferens.     

Relationship of saccular ultrastructure to otolith growth in the teleost Oreochromis niloticus

The sacculus of Oreochromis niloticus is anatomically separated from the utriculus and semicircular canals. The saccular wall is composed of the sensory epithelium, transitional epithelia, and squamous epithelium. Cellular granules are abundant in the sensory and transitional epithelia but scarce in the squamous epithelium. Over the dorsal side of the dorsal transitional epithelium there exists an oval patch of cells with distinctive microvilli. New finding is a shallow groove which extends from the anterior end of the sensory epithelium approximately halfway down along the ventral perimacular transitional epithelium. Small vesicles, which appear “empty” under transmission electron microscopy (TEM), are aggregated in the posterior region of the groove. These small vesicles are also present in both the sensory and transitional epithelia. A second kind of vesicle is comparatively large and appears filled with stainable contents. These vesicles are restricted to the sensory region. Both kinds of vesicles appear to be involved in apical secretion and possibly provide the otolithic membrane with fibers. The otolithic membrane is composed of a gelatinuous layer and subcupular meshwork. The meshwork appears to contribute to the formation of the otolith. The small empty vesicles appear to originate in sensory and transitional epithelial cells and may form the subcupular meshwork. The larger filled vesicles are derived predominantly from sensory cells in the sensory epithelium and appear to contribute to the gelatinuous layer of otoliths. © 1992 Wiley-Liss, Inc.     

Studies on the fine structure of the dorsal vessel of arthropods

Summary The beetle family Altioidae contains some very archaic surviving species, and at the same time, very modern ones. In one lineage from archaic to modern (Fornterita — Alticini — Oedionychini), the number of spermatozoa has been stepwise (according to a geometric series, 2ⁿ) reduced from 256 to 16 per bundle. Comparison with the relatively few data in literature show that this trend is common in insects: more archaic orders have more sperm cells per bundle than the more modern ones, and within an order, the most modern or most specialized groups tend to have least. The reduced number of spermatozoa per bundle may be a sign of reduced sperm production in general, and this probably has adaptive value in limiting genetic variability.A small number of spermatozoa per bundle is sometimes accompanied by an extra large size of spermatocytes. Except for the alticid subtribe Oedionychina, it is hard to see what adaptive value this could have. In the Oedionychines, the large size of the spermatocytes is a prerequisition for the evolution of gigantism of the asynaptic sex chromosomes.Partly supported by a grant (GB-4522) from the National Science Foundation (Washington).I wish to express my gratitude to Prof. Dr. Jan Bechyné, Instituto de Zoología Agrícola, Universidad Central de Venezuela, Maracay, Venezuela, for identification of most alticids mentioned in this paper.     

Ultrastructure of the neurohaemal region of the toad median eminence

Summary According to the internal structure and size of the granules, six types of nerve endings can be distinguished in the toad median eminence: 1. Endings containing mostly dense granules of 600 Å in diameter; 2. Endings containing dense granules of about 800 Å in diameter; 3. Endings which contain dense granules 1,000–2,000 Å in diameter, with the peak at 1,200–1,400 Å; 4. Endings containing granules with a characteristic structure, which differentiate them from the other three types; 5. Scarce endings containing granules 2,000 to 3,800 Å in diameter; and 6. Endings containing only vesicles 400–500 Å in diameter. Types 3 and 4 endings are mainly found in the outer pericapillary zone, and are probably responsible for the strong Gomori-positive reaction observed in this zone. The other four types of endings occur mainly in the inner pericapillary zone, and appear to be Gomori-negative.The probable origins of the different types of endings, and their possible relations with the different releasing factors is discussed.The subendothelial basement membrane has numerous long processes which form a complicated network in contact with all the nerve endings, some nerve fibres and glial cells.Two types of glial cells are described. Pinocytotic vesicles are frequently seen at the points where these cells contact the basement membrane. All the ultrastructural features suggest that these cells are carrying out transport functions.Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina.The author is very grateful to Professor H. Heller for his continued encouragement and criticism and to Mr. J. Lane and Mr. P. Heap for their valuable help.     

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.     

Submicroscopic structure of spermatozoa of a cyprinodontiform teleost,Jenynsia lineata

Summary A submicroscopical study of Jenynsia lineata spermatids and spermatozoa has been made. The formation of an axial nuclear cavity by folding is described. A sub-mitochondrial net with a very geometrical pattern has been found in the mid piece; to the author's knowledge this structure is not comparable to any other described up to now. Taxonomic importance of spermatozoa ultrastructural differences is stressed.This work was supported by a grant of the Consejo National de Investigaciones Científicas y Técnicas of Argentine. Dr. Narbaitz holds a research position and Dr. Dadone a fellowship in the same Institution.     

Estudio y clasificacion de 156 cepas del genero „Candida“ Berkhout, 1923. — Consideraciones sobre candidomicosis, (Moniliasis)

Summary The authors have made an study and an specific classification of 156 yeast-like forms of fungus beloning to the genusCandida isolated in their majority from differents products of the human body.They verify also the high percentage of parasitism with is found theCandida albicans and the roll this specimen has as an aetiological agent in certain vulvo-vaginitis, bronchomycosis, onychomycosis and perionychomycosis, moist mycosis of interdigital spaces, inguinocrural, perianal, axillary and others cutaneous folds.With the name of Candidomycosis are designed the pathological process produced by fungus beloning to the GenusCandida.

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Trabajo presentado a la 4a. Jornada Clínica de Verano en el Hospital Viña del Mar los días 25-26-27-28 de Enero de 1950. (Viña del Mar;Chile).

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Profesor Adjunto de Parasitología de la Facultad de Medicina de Buenos Aires. — Jéfe del Laboratorio del Instituto de Clínica Quirürgica.

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Médico Jefe del Laboratorio de Micología y Tuberculosis del Instituto de Microbiología e Inmunología del ProfesorHugo Vaccaro de la Universidad do Chile.-Becado durante el año 1949 en Argentina por la Comisión Nacional de Cultura, para realizer estudios de Micología con el Prof. Dr. Flavio L. Niño.

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Ayudante del Laboratorio del Instituto de Clínica Quirúrgica.     

Ultrastructural carbohydrate cytochemistry of gastric epithelium

Synopsis On examination with ultrastructural methods for visualizing thevicinal glycols and acid groups of complex carbohydrates, the most superficial surface epithelium of the rat gastric corpus displayed biphasic mucous droplets consisting of a cortex of hexose-rich (i.e. periodate-reactive) neutral mucosubstance and an uncharacterized denser core plus monophasic droplets with the neutral mucosubstance. In many surface epithelial cells of the foveolae, the biphasic and monophasic droplets with the neutral mucosubstance intermingled in varying proportions with monophasic droplets showing uniform periodate reactivity, a variable degree of dialyzed ironbinding—demonstrative of acidic glycoconjugate, and high iron—diamine affinity—demonstrative of sulphomucin. Deep foveolar epithelium displayed only monophasic droplets, most of which contained acidic periodate-reactive complex carbohydrate. Underiying cells, designated isthmus cells, exhibited monophasic or occasional biphasic granules containing sulphated, hexose-rich mucosubstance. Nascent droplets or granules near the Golgi zone differed from the mature organelles in the distribution of the glycoconjugate. Mucous neck cells occupied a deeper stratum and displayed a uniform population of monophasic mucous droplets with a loose meshwork of neutral mucosubstance.Techniques for demonstrating hexoses ultrastructurally stained all Golgi cisternae in the mucigenic epithelium, showing increasing reactivity toward the maturing face. Distinctive cistemae with moderate reactivity in the Golgi complex of isthmus cells were interpreted as GERL. Acidic mucosubstances were visualized only in the inner, mature cisternae of the Golgi complex of cells storing acidic glycoconjugates, and not in cisternae interpretable as GERL.The apical plasmalemma of isthmus cells uniquely exhibited abundant sulphated glycoconjugate and that of parietal cells revealed a less prominent, periodic neutral mucosubstance. Lateral and basal plasmalemmae varied from unstained to slightly reactive; basement membranes showed moderate reactivity with methods for visualizing complex carbohydrates. Abundance of glycogen further characterized surface epithelial cells of the corpus and of some parietal cells     

Estudio micologico de un caso de Micetoma por Cephalosporium en Puerto Rico

Resumen Hemos descrito un Cephalosporium como agente etiológico en un caso de Pié de Madura puertorriqueño. Este hongo crece bien en los medios ordinarios de laboratorio produciendo colonias características y dando origen a un pigmento difusible cuyo color varía de acuerdo con los ingredientes dal agar. Al examen microscópico, los cultivos nos revelan un gran número de conidios alargados que se caracterizan por su gran tamaño, su pronunciado eneorvamiento y su tabicación interna, a más de lo cual se notan muchos clamidosporos de morfología variadísima y, finalmente, estructuras nodulares muy semejantes a las que se desarrollan en ciertas especies del género Trichophyton. La identificatión específica del parásito ha quedado pendiente hasta tanto pueda hacerse un estudio comparativo de otras tres especies cefalospóricas que parecen guardar estrecha analogía con la nuestra a juzgar por las descripciones que de aquellas se han hecho.Sea cual fuere su posición sistemática, la especie aquí descrita no había sido encontrada nunca antes en las lesiones del micetoma. La literatura médica solamente registra otro caso de esa enfermedad producido por Cephalosporium. Se trataba entonces de una infectión contraida en el Brasil que tuvo como agente etiológico en parásito, distinto del nuestro, el cual fué estudiado y clasificado por Leão y Lobo como especie nueva : el Cephalosporium recifei.     

Sur l'ultrastructure des hyphes d'Hypomyces chlorinus Tul. cultivé en présence ou en absence de biotine

Résumé H. chlorinus Tul., Ascomycète parasite des carpophores deBoletus sp. est déficient partiel en biotine.Les caractéristiques ultrastructurales des hyphes obtenues en présence de biotine sont données. Différentes techniques de fixation et de coloration (Thiery, Rambourg) ont été utilisées pour définir la localisation des polysaccharides et l'origine des vésicules apicales. La présence d'un appareil de Golgi chez cet Ascomycète est discutée.La carence en vitamine entraÎne,in vitro, des variations ultrastructurales telle que: l'hypertrophie de certaines cellules; l'épaississement des parois fongiques qui ont un aspect stratifié et qui présentent des irrégularités dans leur épaisseur leur donnant un aspect irrégulier; la mise en évidence moins nette du plasmalemme; la fréquence moins importante des systèmes endomembranaires; la présence d'inclusions lipidiques et de glycogène.Ces observations sont mises en relation avec les résultats de recherches biochimiques que nous effectuons parallèlement. La carence en biotine entraÎne une déviation du métabolisme cellulaire qui s'oriente vers la constitution de réserves cytoplasmiques plutÔt que vers la synthèse de nouvelles structures, ce qui aboutit à un développement réduit du mycélium.

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Summary H. chlorinus Tul., Ascomycete, parasite on carpophores ofBoletus sp., require biotin for optimal growth.The ultrastructural features of hyphae grown on biotin optimal medium are described. Several methods of fixation and coloration (Thiery, Rambourg) are applied for localization of polysaccharides and research of apical vesicles origin. Occurrence of Golgi apparatus in this Ascomycete is discussed.Morphological and ultrastructural changes induced by biotin deficiency consist of: hypertrophy of many cells; thickening of cell walls which appear stratified with variations in their thickness; decreased distinctness of plasmalemma; decreased number of organelles (mitochondria, endoplasmic reticulum); occurrence of numerous lipidic and glycogenic storage granules.These observations on changes in fine structure are related to changes in chemical composition of cells, specially cell walls and to part played by the different organelles in cellular metabolism. This, is oriented to formation of cytoplasmic storage rather than synthesis of new structures leading to a very poor mycelial development.

     

The effect of castration upon the ultrastructure of the rat hypothalamus

Summary The neurosecretory hypothalamic nuclei and the inner zone of the median eminence of castrated rats were studied under the electron microscope. After one month of castration all the neurosecretory neurons of both nuclei show signs of hyperactivity characterized by dilated cisternae of the endoplasmic reticulum containing a macromolecular filamentous material and an increase in the number of ribosomes. After six months of castration, some neurosecretory neurons show an increased number of neurotubules and larger lysosomes than in the controls. Other neurons show a very significant hypertrophy of the endoplasmic reticulum, with large amounts of intracisternal filamentous material. These cells have few neurosecretory granules and in the adjacent synapses the number of granulated vesicles is increased. In the supraoptic nucleus there are two kinds of neurosecretory axons: the clear ones, which are similar to those that appear in control animals and the dark ones, which have smaller elementary granules. In the inner zone of the median eminence the axons show an increase in the number of neurosecretory granules with respect to the controls. After supplementary administration of sexual hormones, all the modifications produced by castration disappear. The ultrastructural changes observed in the neurosecretory nuclei after castration are discussed in relation to those previously described in the neurohypophysis under the same experimental conditions. A feedback regulatory action of sex hormones on hypothalamic neurosecretory neurons is postulated.Supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas and by the Air Force Office of Scientific Research (AF-AFOSR 963-67).We are deeply indebted to Mrs. Defilippi-Novoa and Mr. Alberto Saenz for their skillful assistence.     

The reticular substance of the medulla oblongata of the albino rat

Summary The region of the reticular giganto-cellular nucleus, perfused with formalin and postfixed in osmium tetroxide, was studied with histochemical and electron microscopic techniques. The perikarya of the neurons have two zones. The peripheral cytoplasm contains Nissl bodies, mitochondria, and free RNP particles. The juxtanuclear cytoplasm contains the Golgi complex, mitochondria, RNP particles and dense bodies. The nucleus is indented and has a prominent nucleolus and a paranucleolar body. Dense bodies are found along the axon and dendrites as well. Three different types of synapses are described and two types of synaptic vesicles (spherical and ellipsoidal) are shown.The capillary endothelium shows microvilli and marginal flaps. The endothelial cytoplasm contains vacuoles, micropinocytotic vesicles, and a few dense bodies. Processes of pericapillary cells, surrounded by a basement membrane, also contain dense bodies. The dense bodies found in the neurons and endothelial cells show acid phosphatase activity. On the basis of their morphology and their enzymatic activity these bodies are identified as lysosomes.Partially supported by a school grant No RF 62051 from the Rockefeller Foundation, New York, USA, and Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina.Fellows of the Consejo National de Investigaciones Científicas y Técnicas, Argentina. The authors wish to thank Dr. Mario H. Burgos for his constant interest and criticism, and to Dr. Eduardo Rodriguez Echandia and Dr. Fabio L. Sacerdote for their valuable assistance.     

habitat de Nocardia asteroides,Phialophora pedrosoi y Cryptococcus neoformans en Venezuela

Resumen Se estudiaron 336 muestras de tierra, madera y plantas provenientes de diferentes zonas del Edo. Mérida/Venezuela y Estados vecinos. Fueron inoculados cobayos y hamsters intratesticularmente y ratones por via endovenosa. Testículos y pulmones respectivamente fueron examinados por medio de cultivos e histológicamente.De 11 muestras se aislaronNocardia asteroides, de 6Phialophora pedrosoi y de unaCryptococcus neoformans.Se discuten las probables causas de porqué no se logró aislarParacoccidioides brasiliensis.

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Summary 336 samples of soil, wood and plants from several aereas of the State of Mérida/Venezuela and adjacent regions were studied.The samples were injected into the testes of guinea pigs and into mice intravenously. The testes and lungs of the animals respectively were cultured and examined histologically. From eleven samples were isolatedNocardia asteroides, from sixPhialophora pedrosoi and from oneCryptococcus neoformans.It is discussed, why it was not possible to isolateParacoccidioides brasiliensis.

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Zusammenfassung Es wurden 336 Erd-, Holz- und Pflanzenproben aus verschiedenen Gebieten des Staates Mérida/Venezuela und benachbarten Zonen untersucht. Das Material wurde Meerschweinchen intratestikulär und weissen Mäusen intravenös injiziert. Hoden und Lungen der Tiere wurden auf Nährböden verbracht und histologisch untersucht.Elfmal wurdeNocardia asteroides, sechsmalPhialophora pedrosoi und einmalCryptococcus neoformans isoliert und histologisch nachgewiesen. Es werden die möglichen Gründe diskutiert, warum es nicht gelangParacoccidioides brasiliensis zu isolieren.

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Del Instituto de Anatomía Patológica y Laboratorio de Micología de la Universidad de Los Andes, Mérida/Venezuela y del Laboratorio de Micología, Escuela de Medicina de la Universidad de Cincinnati/Ohio EEUU.

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Este trabajo pudo realizarse con la ayuda económica proporcionada por el Sr. Rector de la U.L.A., Dr.P. Rincón Gutiérrez a quien damos nuestras gracias más sinceras.A Wm. Bridge Cooke, Ph. D. y al Dr.G. A. de Vries agradecemos la identificación de las cepas dePhialophora pedrosoi.

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A los Dres.R. Kohler yP. N. Tablante G., y al Sr.Oswaldo Juergenson nuestro agradecimiento por la ayuda prestada en la recolección de las muestras.     

Light and electron microscopy of granules in the toad choroid plexus

Summary Two types of granules can be distinguished in the toad choroid plexus under the light microscope: pigment granules, mainly localized in the cells that line the free ends of the choroidal villi, and Gomori-positive granules, present in most epithelial cells.The ultrastructural analysis of the choroid plexus reveals three types of granules: multivesicular bodies (MVB), multigranulous bodies (MGB) and dense bodies (DB), and intermediate stages between the last two bodies. The pigment granules seen under the light microscope probably correspond to the DB of the electron micrographs, and the Gomori-positive granules to the MGB. The probable role of these bodies is discussed and so is the significance of the glycogen present in the choroidal cells, their processes and endothelium.This study was partially supported by the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina and the Rockefeller Foundation (School grant RF — 58028).Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas de la República Argentina. The author wishes to thank Prof. M. H. Burgos for his constant interest. His thanks are also due to Prof. H. Heller for providing certain facilities in his department and for his criticism.     

The effects of adenohypophysectomy on the hypothalamic-hypophysial neurosecretory system and the adrenal gland of the toad Bufo arenarum Hensel

Summary The effects of adenohypophysectomy were studied on the hypothalamichypophysial neurosecretory system and the adrenal glands of Bufo arenarum Hensel. An increase in vascularization of the pars intermedia was found and the neurosecretory material (NSM) in the glandular region of the median eminence disappeared. Its reappearance later was accompanied by hypertrophy of the pars intermedia; differentiated chromophil cells appeared in the pars intermedia around the vessels and, at the same time, the adrenal reverted to normal. These findings are interpreted as hypertrophy and differentiation of the pars intermedia with replacement, at least partly, of the functions of the pars distalis. The probable mechanism of differentiation and the functions of chromophil cells of the pars intermedia are discussed.This paper was presented at the VI Reunión de la Asociación Latinoamericana de Ciencias Fisiológicas (A.L.A.C.F.), Viña del Mar, Chile, 1964. It was carried out under the auspices of the Consejo Nacional de Investigaciones Científicas y Técnicas (Argentina) and the Rockefeller Foundation (School grant RF-58028).Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina.The authors wish to thank Prof. M. H. Burgos for his constant interest, Prof. B. A. Houssay, Prof. H. Heller and Dr. J. H. Tramezzani for their criticisms, Miss B. Rodriguez and Mr. L. Castro for their technical assistance.     

On the glycocalyx,the external coat of the plasma membrane,of some secretory cells

Summary The free surface of epithelial cells of secretory organs (human placenta, lactating mammary gland of the rat, choroid plexus of man and rat) and of the accessory organs of the genital tract of the male rat is characterized by a plasmalemmal differentiation named glycocalyx or surface mucous coat. This structure is built up by filamentous or globular substructures.Two main ultrastructural types of the glyeocalyx were observed: 1) The filamentous type such as in the rat epididymis, which resembles the cat intestinal glyeocalyx (Ito, 1965) and that one of human transitional epithelium (Monis and Zambrano, 1968), and 2) The globular type, as observed in the lumen of the lactating mammary gland of the rat.Sialic acid was demonstrated histochemically in the luminal glyeocalyx of all organs studied. In addition, the glyeocalyx of acinar cells of the lactating mammary gland contains sulfate and phosphate groups which were identified by histochemical technics, using enzymatic digestion procedures, suggesting the chemical heterogeneity of this glyeocalyx.Present investigations follow the working hypothesis that the complex carbohydrates of glycocalyces become part of the product of activity of secreting cells.We thank Mr. Luis Iwakawa, Miss Silvia Falcón, Miss Elsa M. Orgnero for technical help, Miss Graciela Aliaga for secretarial assistance. Photography by Mr. H. Magnani. Dr. Hugo F. Carrer cooperated in the initial stages of this investigation.The authors acknowledge the use of the electron microscope of the Department of Pathology, Córdoba University Medical School, for which they thank Prof. E. Mosquera and Dr. E. Hliba. Dr. Hliba photographed picture number 4.     

Ultrastructural aspects of cytoplasmic male sterility inPetunia hybrida

Summary Anther development of isogenic male fertile and cytoplasmic male sterile types ofPetunia hybrida cv. Blue Bedder is studied by electron microscopy. First deviation in sporogenesis of the sterile type, is observed during leptotene stage of the meiocytes. Initial aberration is represented by the presence of large vacuoles in the cytoplasm of the tapetal cells. These vacuoles reveal the first aspects of degeneration; no other ultrastructural differences are observed. Vacuolation is accompanied by the condensation of cytoplasmic organelles. The tapetal cells become distorted and ultrastructural aberrations in mitochondria do occur. The mitochondria elongate and contain several tubular cristae.Substantial evidence suggests, that cytoplasmic male sterility in petunia is encoded by the mitochondrial genome (Boeshore el al. 1983). However, before degeneration becomes manifest, no consistent ultrastructural differences in mitochondrial organization are observed.Abortion of the tapetum and the sporogenous tissue in cytoplasmic male sterile plants, generally follows a corresponding pattern. Ultimately, the cells are highly distorted, the nucleus is disrupted and the cytoplasm disorganized. Mitochondria and plastids degenerate and many lipid droplets are present.     

Saitoella coloradoensis sp. nov., a new species of the Ascomycota, subphylum Taphrinomycotina

Saitoella coloradoensis sp. nov. (NRRL YB-2330, CBS 12360, type strain, MycoBank accession number 563858) is described. This new member of the phylum Ascomycota, subphylum Taphrinomycotina was isolated from insect frass occurring in an Engelmann spruce (Picea engelmannii) that was growing in Colorado, USA. Multigene sequence analysis showed that S. coloradoensis is distinct from Saitoella complicata, the only other known species of Saitoella. The two species may be separated phenotypically from growth reactions on d-xylose, ribitol and methyl-α-d-glucoside. Asexual reproduction is by budding and both species produce thick-walled, spherical cells that appear morphologically similar to the ascogenous cells formed in plant host tissue by species of Protomyces and some species of Taphrina. The thick-walled cells did not form ascospores but did produce buds when placed on fresh growth media.     

Cytochemistry of 5-hydroxytryptamine at the electron microscope level

Summary The nerves of rat pineal gland are known to contain norepinephrine and 5-hydroxytryptamine. With the glutaraldehyde-dichromate reaction for the cytochemical localization of unsubstituted catechol- and indoleamines, dense reactive granules could be demonstrated in such endings. A similar reaction was observed in the adrenergic nerves supplying the vas deferens and storing exclusively norepinephrine. Formaldehyde fixation, prior to the glutaraldehyde-dichromate treatment, interferes with the reaction given by catecholamines not affecting the indolic reactive sites. After this combined procedure pineal nerves still exhibited the dense reactive granules, while these were not found in the nerves of the vas deferens. Following bilateral cervical sympathectomy reactive granules disappeared from the perivascular processes of the pineal gland. No reaction could be observed in the cytoplasm of parenchymal cells neither in their perivascular processes.These cytochemical results suggest that both catecholamines and 5-hydroxytryptamine are contained within the granulated vesicles of pineal nerves.This work has been supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina and U.S.Air Force (AF-AFOSR 963-66).Fellow of the Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina. We want to express our gratitude to Prof. E. De Robertis for his constant help and encouragement; and to Miss Nélida Fernández Oranges, Mr. Raúl Castelli and Mr. Alberto Sáenz for their skillful technical assistance.