The fine structural localization of endogenous and exogenous peroxidase activity in human bone marrow mast cells under pathological conditions

We have examined the ultrastructural characteristics of peroxidase activity in human bone marrow mast cells. These studies were performed in three patients with systemic mast cell disease, and in another six patients showing bone marrow mast cell hyperplasia. Endogenous peroxidase activity was localized in the perinuclear cisternae and strands of endoplasmic reticulum, but never in the granules. We have also demonstrated the "in vivo" existence of exogenous peroxidase activity in two of the three cases of systemic mast cell disease. The peroxidase internalization involved its binding to the plasma membrane, followed by its incorporation into the cell by a general endocytic process comprising the uptake of dispersed peroxidase-positive material mainly by phagocytosis of granular structures containing peroxidase. The exogenous peroxidase appeared in non-membrane bound granules, vacuoles or aggregates, but we have never seen the enzyme linked to the mast cell granules.     

On the presence of conspicuous electron dense bodies in the pinealocytes of the pig

Summary The pinealocytes of the pig contain a large number of aldehydefuchsin positive granules. In order to determine their nature an ultrastructural study was carried out. Numerous bodies having a maximal diameter of about 1,600 nm were found. These elements showed a great variety of internal structure, ranging from a homogeneous content and amorphous dense aggregates to lamellate bodies. Although only a few of them displayed a positive reaction for acid phosphatase, their morphological appearance strongly suggests that they belong to the lysosomal system.Aided in part by a grant from the Consejo Nacional de Investigaciones Cientificas y Técnicas de la República ArgentinaAssociate Professor of Embryology and Histology and Member of the Investigator Career of the CONICETProfessor and Chairman of Histology and Member of the Investigator Career of the CONICETCenter sponsored by the Universidad Nacional de La Plata and the Comisión de Investigaciones Cientificas de la Provincia de Buenos Aires     

Analisis de 6000 especímenes micológicos Sintesis estadística. — Comentarios

Resumen En este trabajo se estudia en la Parte I todo el material procedente de especímenes clínicos.En primer lugar se estudian las cepas aisladas de Micosis Superficiales o Dermatomicosis.En segundo lugar, los hongos levuriformes del género Candida procedentes de materiales clínicos varios.En tercer lugar los agentes de Micosis Profundas, y finalmente los hongos del aire de Caracas que tienen interes alergológico.Los resultados de todo aparecen en los cuadros sinópticos adjuntos.

---

Summary In this part of our work we deal with fungi of clinical interest.Firstly, we study specimens isolated from dermatomycoses.Secondly, yeast-like fungi of the genusCandida of clinical origin.Thirdly, fungi producing systemic mycoses and lastly, air-borne fungi of allergic interest from Caracas and its surroundings.

---

Résumé Dans la première partie de ce travail, on étudie tout le matériel d'intérêt médical.En premier lieu, les souches isolées de matériel clinique de Mycoses Superficielles ou Dermatomycoses.En deuxième lieu, les champignons lévuriformes du genre Candida provenant de matériaux cliniques variés.En troisième lieu, les Agents de Mycoses Profondes et, finalement, les champignons contenus dans l'air de Caracas intéressant la Mycologie Allergique.

---

---

(Trabajo de la Sección de Micología del Instituto Nacional de Higiene)

---

Trabajo presentado al IV Congreso Venezolano de Tisiología y Neumonología, Valencia, Diciembre 1959.

---

Jefe de Sección Micología. — Instituto Nacional de Higiene. Jefe de Sección Micología.-Hospital Universitario. Profesor de la Catedra de Micología.-Escuela de Bioanálisis Fac. de Medicina U C V.

---

Directora de la Micoteca Nacional.-Seccion de Micología, Instituto Nacional de Higiene.     

Electron microscopic demonstration of metals in rat mast cells

Summary This paper describes a modification of a cytochemical method for the demonstration of heavy metals. The well localized precipitate in the mast cell granules, which is also present in granules that have been separated from the cell, suggests that the metals are localized in the granules. It is demonstrated that mast cell grown cultures do not contain precipitate. The chelating and histamine inhibiting agent 8-hydroxyquinoline produced no changes in the histochemical pattern of the mast cell granules before nor after treatment with the histamine liberator 48/80 which provokes a release of granules from the cells. These observations suggest either that the metal (zinc) is bound to the granules in such a manner that the chelating agent cannot chemically, or based on the configurations of the metal-containing molecule, reach the metal and theraby prevent its transformation to a metal suphide.     

Ultrastructure of STH cells of the pars distalis of castrated male mice after hepatectomy

Summary An electron microscopic study aimed at differentiation between castration gonadotrophs and posthepatectomy STH cells was performed on the pars distalis of the pituitary of castrated male mice, after partial hepatectomy. The ultrastructural features observed permit the distinction of both cell types. In the present experiments some remarkable ultrastructural changes, other than those described in a previous report, have been found in STH cells of hepatectomized mice with or without previous castration. Most of them contained masses of heterogeneous electron density, suggesting fusion of granules. These masses and some secretion granules were observed close to the plasma membrane, apparently in the process of discharging material into the pericapillary space. Granular extrusion was more frequent than normally. An increased number of lysosomes, probably related to the digestion of overproduced secretion material, was evident. The appearance of concentric lamellar formations might be related to an increase in cell movements. STH cells with severe cytoplasmic damage were also found, indicating an increased rate of cell loss.Work carried out with the financial assistance by a grant from the Comisión de Investigación Científica de la Universidad Nacional de La Plata. Thanks are due to the members of the Technical Staff of the Institute for their technical assistance.     

Ultrastructure of the rat posterior pituitary gland and evidence of hormone release by exocytosis as revealed by freeze-fracturing

Summary Posterior pituitary glands from normal rats, and rats which had been deprived of water for varying periods, were examined by the freeze-fracture method. This technique reveals large areas of the nerve cell membrane. Images consistent with exocytosis as the mechanism of release of the neurohypophysial hormones were observed. These modifications were most numerous after the rat had been starved of water for 2 days.In normal rats, the large number of neurosecretory granules within the nerve fibres caused a bulging of the nerve cell membrane. The bulges disappeared 2 days after removal of drinking water. Regions of the membrane displaying bulges were characterised by the absence of the typical membrane-associated particles.It is postulated that the close proximity of the neurosecretory granules to the nerve cell membrane may result in rapid fusion of the neurosecretory granules on stimulation of the gland. The change in properties of the nerve cell membrane overlying the neurosecretory granules, as suggested by the loss of membrane-associated particles, may represent a change in the structure of the membrane to a form which is more favourable for fusion.This work was supported in part by a grant from the New Zealand Medical Research Council.     

Heterogeneity of the hemoglobin of the Ohrid trout (Salmo L. typicus)

We have analyzed the hemoglobins of five individual trout from the Ohrid Lake (Salmo L. typicus) by electrophoretic methods, by reversed-phase high-performance liquid chromatography, and by limited structural analyses. The two major classes of hemoglobin are type I (35% of total) and type IV (65%). Type IV is the major oxygen-transporting hemoglobin; it consists of three types of chain (in about equal quantities) and three types of chain (one major and two minor types). Several structural differences have been observed between these three (IV) chains and between the three (IV) chains, suggesting a complex genetic system governing the synthesis of these proteins. Moreover, a few amino acid substitutions occur at positions involved in contacts between chains, which suggests that differences in oxygen affinity may exist between these various type IV hemoglobins. Type I hemoglobin is less complex because it contains one type of chain and two chains; the latter two differ in numerous positions, suggesting duplications of the (I)-globin gene. The and chains of type I hemoglobin differ considerably from the and chains of type IV hemoglobin, indicating the existence of (I)- and (I)-globin genes separate from the (IV)- and (IV)- globin genes.This study was supported in part by the Yugoslav-American Joint Funds, pp 812 (to G.D.E.), and by United States Public Health Service Research Grant HLB-05168 (to T.H.J.H.).     

Effects of isoproterenol on the dense core and perigranular membrane of atrial specific granules

Summary Following subcutaneous injections of isoproterenol hydrochloride (ISO), atrial cells present a large number of partly degranulated or completely clear specific granules enclosed by an intact membrane. Such profiles were never encountered in normal controls and might suggest ISO-induced release of a secretory product. Permeability of perigranular membrane was tested using the extracellular macromolecular tracer horseradish peroxidase (HRP). Reaction product was entirely absent within granules of atrial cells in which the sarcolemma was made permeable to HRP molecules by the ISO injections. This seemed to be the case even in heavily labelled cells in which the peroxidase had penetrated the mitochondrial membranes. In atrial cells impermeable to the tracer, the specific granules closely apposed to the sarcolemma were always HRP-negative. The release mechanism of a possible secretory substance from the specific granules is discussed.     

Plant regeneration from somatic embryos ofTaxus brevifolia

Summary Taxusbrevifolia is the source of paclitaxel (Taxol®), an anticancer drug. A method for regeneration ofTaxus brevifolia from immature zygotic embryos via somatic embryogenesis is described. Embryogenic callus tissues were obtained by culturing immature zygotic embryos on Lloyd and McCown medium (MCM) supplemented with 160 M 2,4-dichlorophenoxyacetic acid (2,4-D) + 5 M benzylaminopurine (BA) + 5 M naphthaleneacetic acid (NAA) for 4 weeks. Putative embryoids were obtained following transfer of cultures to MCM medium supplemented with 4 M BA + 5 M kinetin + 1 M NAA for 6 to 8 weeks. Conversion of embryos was obtained on MCM medium supplemented with 40 M abscisic acid (ABA) + 1% activated charcoal. Development of bipolar structures with recognizable shoot and root apices was observed in somatic embryos. Five percent of somatic embryos were regenerated into plantlets on half-strength growth regulator-free MCM medium.     

Localisation ultrastructurale de l'activite de la phosphatase alcaline dans le foie foetal humain

Résumé Les auteurs, dans le cadre d'une étude ultrastructurale du foie foetal humain, ont utilisé une méthode directe à pH 8 pour mettre en évidence l'activité de la phosphatase alcaline.L'accumulation du produit final de la réaction histochimique (phosphate de plomb) a pu être observée de façon constante sur les membranes plasmatiques des hepatocytes et des érythroblastes. Dans la lignée érythropoïétique les dépôts sont toujours plus intenses sur les cellules les plus immatures.Les auteurs ont observé également des dépôts importants de phosphate de plomb dans certains granules péribiliaires ainsi que dans des granules des cellules érythropoïétiques.La disposition des dépôts le long des membranes cellulaires et dans certains organites cytoplasmiques est discutée en fonction de la technique de mise en évidence.

---

Summary A direct lead method for visualization of alkaline phosphatase activity at pH 8 was used in an electron microscope study of the foetal development of human liver.An accumulation of lead phosphate, the final product of the enzymatic reaction, was observed on the cell membranes of hepatocytes and also on those of erythroblasts, especially in immature cells.An intense reaction was observed in peribiliary granules and in granules of the erythropoietic cells.The localization of lead phosphate precipitate along the cell membranes and in some cytoplasmic bodies is discussed in relation to the procedure used.

---

---

Ce travail a été effectué avec l'aide du Fonds National Suisse de la Recherche Scientifique .     

Electron microscopic observations on a new cell type in the fundus mucosa of the mouse stomach

Summary In the course of electron microscopic investigations of the fundus mucosa of the mouse stomach, a few cells of an unknown type were found by chance in the deep portions of the glands. These cells are characterized by two different kinds of specific granules in their cytoplasm, one of which being large and less dense, and the other one being small and dense. The large less dense granules resemble zymogen granules of the chief cell, which are formed by the rough endoplasmic reticulum and Golgi system. The small dense granules are quite similar to the secretory granules of the basal granulated cell, and are considered to be formed in the Golgi complex. Release figures of the small dense granule were not observed, numerous granules, however, were observed in close contact with the basal cell membrane. The occurrence of these two kinds of granules in one cell suggests that the basal granulated cell and the zymogenic cell originate from the same entodermal stem element.The author cordially thanks Professor Dr. Hisao Fujita, Department of Anatomy, Hiroshima University School of Medicine, for his kind advices and criticisms.     

An explication of ‘wind illness’ in Northern Thailand

Wind illness is a very common complaint among the Northern Thai, yet is rarely recognized by Thai physicians trained in biomedicine. Persons most susceptible to wind illness are adult women who have ever borne a child. Consequently, data were obtained from 415 everparous women, 43% of whom reported ever having had wind illness and 57%, never having had it. In addition, 20 individuals who had ever had the syndrome were followed for case study, and 13 indigenous healers who traditionally treat clients suffering wind illness were interviewed. Their perceptions of the etiology, symptomatology and treatment of wind illness are reported in Part I. Part II is an attempt to define wind illness in terms of biomedicine and as a consequence of fertility. Part III synthesizes the emic and etic accounts with explanations for the perdurance of wind illness despite the advances of biomedicine and the recent fertility decline in Northern Thailand.     

Ultrastructural development of the neurohemal organ joined to the ecdysial gland after imaginal moulting in the male isopod Sphaeroma serratum fab. (Crustacea Flabellifera)

Summary The present ultrastructural study deals with the lateral cephalic nerve plexus of Sphaeroma serratum, a neurohemal organ joined to the Y organ (ecdysial gland). This plexus acts as a storage centre for neurosecretory products from two sources: the two autochtonous cells (plexus cells) within the plexus itself, and the neurosecretory cells in various parts of the central nervous system, particulary the mandibular ganglion (A-cells).In prepuberal animals, plexus cells and subesophageal A-cells produce neurosecretory granules of two types measuring 1550±50Å and 1570±40Å respectively. Five categories of axon terminals were distinguished in the plexus. The granules found in two of these terminal types are believed to come from the plexus cells and from the mandibular ganglion A-cells.Cessation of production of neurosecretory granules in these A cells and plexus cells was observed in puberal animals, in the plexus with concomitant depletion and disappearance of different granule categories. The first axon terminals affected by this process are the two categories containing granules originating in the plexus and mandibular ganglion A-cells. Degeneration of the ecdysial gland in male Sphaeroma serratum might be connected with the cessation of granule formation in these two types of cell.

---

Résumé Chez Sphaeroma serratum, la mue de puberté est suivie d'une dégénérescence de l'organe Y (glands de mue). Le plexus nerveux céphalique latéral, organe neurohémal accolé à cette glande a été l'objet de la présente étude ultrastructurale. Cet organe représente un centre de stockage de neurosécrétions qui proviennent d'une part, de deux cellules autochtones (cellules plexales) situées au sein même de ce plexus, d'autre part, de cellules neurosécrétrices situées dans le ganglion mandibulaire (cellules de type A).Chez les individus pubères, les cellules plexales et les cellules A du ganglion sous-oesophagien synthétisent des granules de neurosécrétion dont la taille est respectivement 1550±50Å et 1570±40Å. Il a été reconnu au sein du plexus 5 catégories de terminaisons dont les granules proviendraient pour deux d'entre elles des cellules plexales et des cellules A du ganglion mandibulaire. Chez les animaux pubères on observe un arrêt de la synthèse des granules de neurosécrétion au sein des cellules plexales et des cellules A du ganglion mandibulaire. Simultanément on enregistre dans le plexus la raréfaction puis la disparition des divers types de granules. Ce processus atteint en premier les terminaisons correspondant aux cellules plexales et aux cellules A du ganglion mandibulaire. La dégénérescence de la glande de mue chez les mâles pourrait être en relation avec l'arrêt de synthèse de ces cellules.

     

Stereoscopic back-scattered electron imaging of silver-stained proteins in nucleoli

By using simultaneously the AgNOR silver staining method, back-scattered electron imaging mode and stereo-tilt in scanning electron microscopy (SEM), it is possible to observe the nucleus through the cell surface, the nucleolus, and the tri-dimensional distribution of the AgNOR-associated acidic proteins. In C3H10T1:2 cells and their 7-12-dimethylbenz--anthracene-treated transformants, the staining demonstrates several intranucleolar silver-staining granules (SSG), surrounded by a weakly staining region. The SSG may represent the fibrillar center (FC) and the weakly staining region, the fibrillar dense component (FD). This component can link several SSG together to form a rope-like structure. In cells with no visible nucleolus and inactive nucleolar organizer regions (NORs) the silver-staining granules are less numerous, close together and the presumed fibrillar dense components are not visible. The SSG are located more peripheraly, and the weakly staining region and the rope-like structure are less prominent in control cell nucleoli than in transformed cells with a comparatively high rate of RNA synthesis.     

Discrimination of LH,FSH, TSH and ACTH in dissociated porcine anterior pituitary cells by light and electron microscope immunocytochemistry

Summary Using the PAP unlabelled antibody method, LH, FSH, TSH and ACTH were localized at light microscope level in cultured cells dissociated from the porcine adenohypophysis. Antisera were shown to be specific for the subunits of the porcine glycoprotein hormones by radioimmunoassay and absorption studies. Using these antisera, it was found that LH and FSH were contained within the same cell, with TSH in a separate cell. When absorbed with LH, anti-porcine ACTH stained a separate distinct population of ACTH cells.Adjacent ultra-thin sections stained with anti-pLH and anti-pFSH, and examined at electron microscope level, showed that the ovoid, 150–400 nm secretory granules of the LH/FSH gonadotrophs contained both LH and FSH.The authors gratefully acknowledge the technical assistance of Carole Smith and Adrian Walsh     

Mast cells in human bile duct obstruction

Surgical biopsy specimens obtained from 50 patients with secondary cholangitis caused by obstruction of the common bile duct were studied immunohistochemically. Data on the number and ultrastructural appearances of mast cells positive for tryptase, chymase, vasointestinal polypeptide (VIP), and substance P (SP) were obtained. The bile ducts from patients presenting combined chronic exacerbated cholangitis and chronic sclerotic cholangitis showed significantly higher numbers of mast cell types compared to the controls (P < 0.0001). Cases with sclerotic cholangitis alone had significantly lower number of cells than patients with chronic exacerbated cholangitis alone (P 0.0001). Morphometric measurements of electron micrographs showed that mast cell granules containing VIP, SP and chymase were commensurable in size. Electron-lucent granules without reaction product (altered granules) and granules with focal distribution of the reaction product were observed in all types of mast cells. Furthermore, some nerve fibers positive for SP and VIP and serotonin-positive endocrine cells were observed in close proximity to the mast cells. In conclusion, the results of our study demonstrate the existence of different populations of mast cells, nerve structures and endocrine cells in the lower part of the human large bile duct, and suggest their participation in the development of pathological processes.     

Five new species of Eimeria (Apicomplexa: Eimeriidae) from lizards

Five new species of Eimeria are described from lizards. Eimeria baltrocki n. sp. was found in the berber skink, Eumeces schneideri, from Egypt. The oöcysts are cylindroidal, averaging 38 × 18.3 m, with a single thick oöcyst wall. Most oöcysts possess a single polar granule; a micropyle and oöcyst residuum are absent. The sporocysts are ellipsoidal and average 11.5 × 8.1 m, each with a large, globular sporocyst residuum; the Stieda body is absent. Eimeria anolidis n. sp. is described form the common anole, Anolis carolinensis, from Florida, USA. The oöcysts are cylindroidal and average 31 × 15.8 m with a thick, single-layered oöcyst wall. Two polar granules are usually present; a micropyle and oöcyst residuum absent. The sporocysts are ellipsoidal and average 9.4 × 7.5 m with a globular sporocyst residuum; the Stieda body is absent. Eimeria guyanensis n. sp is recorded in the ameiva, Ameiva ameiva, from Guyana, South America. The oöcysts are spherical to subspherical, average 19.0 × 17.8 m and possess a thick, single-layered oöcyst wall. Numerous polar granules are present (n > 5); a micropyle and oöcyst residuum are absent. The sporocysts are spherical to subspherical, average 7.5 × 7.8 m and possess a compact globular sporocyst residuum; the Stieda body is absent. Eimeria phelsumae n. sp. was recovered from the giant day gecko, Phelsuma madagascariensis grandis, from Madagascar, which harboured a simultaneous infection of E. brygooi. The oöcysts measured 32 × 15 m and are cylindroidal without polar granules, a micropyle or oöcyst residuum, or a Steida body. The sporocysts are ellipsoidal and average 9.8 × 7 m, with a loosely clumped, granular sporocyst residuum; the Steida body is absent. Eimeria leiocephali n. sp. was discovered in the faeces of the ornate ground iguana, Leiocephalus barahonensis, from Haiti. The oöcysts are spherical to subspherical, 21 × 19 m, and contain a number of polar granules (n > 5); a micropyle and oöcyst residuum are absent. The sporocysts are spherical, 8 m in diameter and lack a sporocyst residuum. Eimeria turcicus and E. lineri were found in faeces of Hemidactylus turcicus turcicus from the host's country of origin, Turkey.     

Cytokinesis in the green alga Eremosphaera viridis: Plasmalemma formation from “open membranes”

Summary Cytokinesis in the unicellular chlorococcalean alga Eremosphaera viridis de Bary has been investigated by electron microscopy of thin sections. The new plasmalemmata of the daughter cells in this organism form centrifugally within a phycoplast. Unlike other cell division systems each new plasmalemma is formed, not by the fusion of vesicles, but rather by the fusion of open membranes which are characteristically heavily stained. Measurements of these open membranes reveal that they are 11 nm thick with a central 4,5 nm unstained portion. The possible origin of these open membranes as burst-open vesicles has been suggested from the presence of intensely straining vesicles in the vicinity of the cell equator. Calculations of vesicle and open membrane surface areas support this contention.     

Erratum: Cloning and sequencing of the phycocyanin gene from Spirulina maxima and its evolutionary analysis

The genes were cloned for the two apoprotein subunits, and ,of phycocyanin from the cyanobacterium Spirulina maxima = Arthrospiramaxima) strain F3. The - and -subunit gene-coding regionscontain 489 bp and 519 bp, respectively. The -subunit gene is upstreamfrom the -subunit gene, with a 111-bp segment separating them.Similarities between the -subunits of S. maxima and nine othercyanobacteria were between 58% and 99%, as were those between the -subunits. The maximum similarity between the - and -subunits from S. maxima was 27%.     

A proposito del llamado micetoma maduro-micosico del pulmon

Conclusiones Se efectúa el estudio de 5 observaciones del llamado Micetoma maduromicósico de pulmón en sus aspectos, histopatológico, micológico y clínico.Todas ellas pertenecen a mujeres y configuraron el cuadro de cavidad bronquial empastada, anotando el predominio de su localización en el lóbulo superior izquierdo.Se señala la uniformidad de los caracteres morfológicos que presenta la masa miceliana llamada grano en todos los casos estudiados, en los cuales no fué posible individualizar la existencia de órganos de fructificación que permitiéran una clasificación, cuando más no fuera, genérica del hongo observado.Se critica la aplicación del término Micetoma para éstos casos con igual criterio que el clásico, que supone una enfermedad micótica primitiva, razón por la cual se prefiere hablar de cavidad con contenido micótico o maduromicótico.En el único caso que se logró cultivar al hongo parásito, el estudio micológico del mismo permitió aislar una especie del GéneroAspergillua con caracteres morfológicos sumamente atípicos.

---

Summary The author presents the study of five observations of the so-called Maduromycosis-mycetoma of the lungs in their clinical, histopathologic and mycologic aspects.The pathologic features in all these cases have been found in bronchial cavities of women, situated in the upper lobe of the lung. A compact mycelial mass, the grain, filled up these cavities.In one of the five cases a fungus was cultivated which was classified as belonging to the GenusAspergillus Michelii, with abnormal and atypical features.Short criticism is presented about the concept Maduromycosis mycetoma of the lung, applied by authors designating this process.