Sarcoma arising from interdigitating cells. Cytology and cytochemistry

The cytologic features of a highly malignant sarcomatous tumor in a 37-year-old male, arising from interdigitating cells and localized in the mediastinum, lymph nodes and skin, are described. Cytologically this sarcoma was characterized by large cells with ill-defined, faintly basophilic cytoplasm, monocytoid or multilobulated nuclei and a reticular chromatin structure; very prominent nucleoli were seen in some of the cells. Some of the tumor cells were spindle shaped. The ultrastructurally characteristic invaginations of the cell membrane were not obvious in the cytologic smear, although the nuclear membrane showed deep, narrow, channel-like indentations. The specific ultrastructural, immunologic and cytochemical characteristics of the interdigitating cells were recognized in the tumor cells. Adenosine triphosphatase was present in the tumor cells in large amounts, while acid phosphatase, acid alpha-naphthyl acetate esterase and other enzymes were absent. The described tumor must be considered another tumor of the mononuclear phagocyte system; the proposed name is "interdigitating-cell sarcoma."     

四种淡水养殖鱼类血细胞的细微结构

四种淡水鱼的血细胞形态基本相似。红血球形态与其他低等脊椎动物基本相似。淋巴球绝大部分是小淋巴球:单核球数量较少;四种鱼的嗜中性白血球形态结构差不多,胞核多为蚕豆形,很少见分叶核,分叶一般也只有二叶,这与哺乳类显然不同;嗜酸性白血球的形态结构与其他脊椎动物基本相似;在少数血涂片中看到了嗜碱性白血球。       

A STUDY OF GLUCO-CORTICOSTEROID-INDUCED PYKNOSIS IN THE THYMUS AND LYMPH NODE OF THE ADRENALECTOMIZED RAT

Pyknotic nuclei, observed in the thymus of steroid-treated rats, are dense, homogeneous, intensely basophilic and Feulgen positive. Under the electron microscope, the image is that of a complete segregation of the chromatin from the nuclear sap producing a margin or crescent of condensed chromatin. Approximately 30% of all small thymocytes appeared to undergo this type of degeneration within 3–4 hr after administration of the synthetic corticosteroid, dexamethasone. At this time, pyknotic thymocytes were observed in clusters, probably as a result of the activity of dense reticular cells and macrophages. Topographical and experimental data suggest the existence of a select population of steroid-sensitive thymic cells. Furthermore, on the basis of thymidine-³H incorporation studies, it appears that the steroid-sensitive population of thymocytes does not correspond to "aged" cells. In addition, many plasma cells became pyknotic after the same steroid treatment, indicating an unexpected similarity between their nuclei and those of lymphocytes. Finally, steroid failed to induce pyknosis of thymocytes in a variety of in vitro experiments, suggesting that the in vivo effect of steroid is of an indirect nature. The results are discussed in terms of (a) the nature of the nuclear changes characterizing pyknosis, (b) the hypothetical mechanism whereby steroids trigger such changes, and (c) the population of cells susceptible to steroid-induced pyknosis.     

OBSERVATIONS ON THE STRUCTURE OF RHODOSPIRILLUM MOLISCHIANUM

The lamellae of the bacterium Rhodospirillum molischianum originate as extensions of the cytoplasmic membrane into the cytoplasm of the cell. Initially, these extensions are narrow folds and occur independently of one another. The first lamellae to appear average about 80 A in width, representing one side of the infolded cytoplasmic membrane, or 160 A when the two sides of the fold are closely appressed. The 160-A lamellae increase in number and may associate to form larger lamellae, which represent varying degrees of association between adjacent folds. Later, the space within each fold increases; the two appressed regions of the cytoplasmic membrane in each fold separate to form distinct invaginations, and the lamellae observed at this stage are formed by an association of the sides of adjacent invaginations.     

A simple method for the fluorescence analysis of nucleic acid-dye complexes in cytological preparations

This communication describes a simple method for recording fluorescence emission spectra of cytological preparations using a conventional fluorescence spectrophotometer. The emission characteristics of "in situ" complexes between some basic fluorochromes (DAPI, 33258 Hoechst, acridine orange, pyronin Y, and ethidium bromide) and nucleic acid containing structures from smears of chicken blood and Ehrlich tumor cells (chromatin, basophilic cytoplasm) are briefly described.     

The Histochemical Demonstration of Hemoglobin in Blood Cells and Tissue Smears

The demonstration of intracellular hemoglobin in permanent preparations has long been a problem. The affinity of hemoglobin for iron hematoxylin is well known but this stain also colors yolk, chromatin, and other structures and is therefore not a reliable criterion. The presence of hemoglobin has been associated with an acidophil cytoplasm which stains a characteristic color, but a careful inspection of living cells in early hematopoetic or embryological stages demonstrates that hemoglobin is present in the erythrocytes which are quite basophilic. In the course of some research on the blood of embryonic frogs it became desirable to demonstrate the presence of hemoglobin in cells by means of a specific staining reaction.     

FORMATION OF GERM CELL-LIKE CELLS IN THE PERITONEAL EPITHELIUM OF THE TESTES OF ESTROGEN-TREATED ADULT MALES OF THE JAPANESE RED-BELLIED NEWT, CYNOPS PYRRHOGASTER PYRRHOGASTER

Columnar cells of the peritoneal epithelium in slender cords of the testes were examined in normal and estradiol benzoate-treated Japanese red-bellied newt, Cynops pyrrhogaster pyrrhogaster, by light and electron microscopy. In normal newts, the peritoneal epithelium covering the slender cord consists of columnar cells, which contain extraordinarily large, oval or spindle-shaped nuclei with conspicuous indentations. The nucleus contains chromatin granules and the cytoplasm is filled with numerous tonofilaments. The primordial germ cells are scattered throughout the slender cord, and each cell is surrounded by a few follicle cells. Between the germ cells and follicle cells there are microvilli-like processes. The nucleus of primordial germ cells is multilobate and has electron lucent areas, dispersed chromatin and several electron-dense nucleoli. In the lighter cytoplasm, the nuage material is found very near to nuclear pores, and is frequently seen among the mitochondria. The nucleolus-like body is not associated with other organelles. The primary spermatogonia have bilobate nuclei. It is remarkable that most of the cytoplasmic organelles are found in the deep nuclear indentations. The nuage material and nucleolus-like body are well developed in the cytoplasm. After treatment of newts with estradiol benzoate for one year, four types of cells can be distinguished in the peritoneal epithelium. One type is quite different from the columnar cells. These newly appeared cells are large and light in appearance. Their nucleus is highly lobate, and contains dispersed chromatin and several nucleoli with compact electron dense material in its periphery. The cells are characterized by the presence of nuage material and nucleolus-like bodies in the cytoplasm. There are microvilli-like processes between these cells and adjacent elongated cells. These ultrastructural characteristics of the light cells are very similar to those of primordial germ cells and/or primary spermatogonia in normal testes. These findings suggest that the light cells which appear in the peritoneal epithelium of the testes on administration of estrogen may be germ line cells.     

Substance P innervation of the rat thymus

Immunohistochemistry for substance P (SP) in the rat thymus revealed fine varicose neural profiles in specific regions of the thymus. Thymic SP innervation was abundant within the capsule and interlobular septa. The majority of SP+ nerve fibers within the septa were free of vascular association, although some fibers were associated with the vasculature deep within the septa. SP+ nerve fibers entered the thymic cortex from the septa and distributed among cortical thymocytes and mast cells. Along the corticomedullary junction, SP+ nerve fibers were found in association with the vasculature. The medullary region of the thymus received only a sparse innervation of SP+ fibers. In addition, SP+ nerve fibers coursed adjacent to OX-8+ cells and mast cells in the extrathymic connective tissue surrounding the thymus. The present study provides evidence that SP is present in nerve fibers in the thymus, and may be available to interact with thymocytes, mast calls, and other cells in the thymus, and affect their development and function.     

甜菊愈伤组织中的质膜内陷:超微结构和酸性磷酸酶细胞化学定位

对在分化条件下的甜菊 (Stevia rebaudiana)愈伤组织分生区域细胞的质膜内陷进行了超微结构和酸性磷酸酶细胞化学研究。结果表明 ,在不同液泡化状态的细胞中均有质膜内陷存在。在原生质浓密的细胞中 ,质膜呈起伏的波纹状 ,某些部位发生明显内陷 ,大小不等 ,多呈圆球状。在部分液泡化细胞中 ,质膜内陷体积增大 ,内含物增多且结构复杂。在液泡化细胞中 ,质膜内陷嵌入中央液泡 ,但彼此间以一膜间隙隔开。质膜内陷中的内含物以小泡和卷绕的膜结构形式存在。酸性磷酸酶活性定位结果显示 ,质膜及其内陷含高的酶活性。推测质膜内陷在功能上与液泡相似 ,构成了这些细胞水解空间的一部分。     

STUDIES ON THE ULTRASTRUCTURE OF THE GUARD CELLS OF OPUNTIA

The guard cells of Opuntia contain numerous mitochondria, elements of endoplasmic reticulum, dictyosomes, and microbodies. A complex array of small to large vacuoles which contain small, membrane-bounded vesicles occur in each guard cell. The variety of cytoplasmic constituents and vacuoles suggest that the guard cells are complex in function. A highly reduced grana-fretwork system within the plastids indicates that the photosynthetic capacity of the guard cells is probably rather low. No plasmodesmata occur in the walls between the guard cells and the subsidiary cells while there are numerous invaginations of the guard cell plasmalemmas. Many of the variations in the plasmalemma probably indicate that the plasmalemma is a highly active interface.     

Are any functionally mature cells of medullary phenotype located in the thymus cortex?

Experiments were undertaken to test if thymocytes of "mature" or "medullary" phenotype were restricted to the medullary area of the thymus. A calculation based on direct cell counts on serial sections indicated that 11.5% of adult male CBA thymic lymphoid cells were within the medullary zone. Since only 3-4% of thymocytes were cortisone resistant, the majority of thymocytes within the medulla were, like cortical thymocytes, cortisone sensitive. A series of cell surface antigenic markers, used alone or in pairs, suggested that 13-15% of thymocytes were of medullary phenotype, somewhat more than the number of thymocytes actually present in the medulla. However, much of this discrepancy could be explained by differential death of cortical cells during isolation and staining, and by the existence in the cortex of a subpopulation of early blast cells which shared some, but not all markers with medullary thymocytes. A direct test for mature or medullary phenotype cells in the cortex involved selective transcapsular labeling of outer-cortical cells with fluorescent dyes, followed by multiparameter immunofluorescent analysis of the 10% labeled population. Outer-cortical thymocytes included some cells (mainly early blasts) sharing some markers with medullary thymocytes, but very few (less than 1%) of these cells expressed all the characteristic "mature" markers. Limit-dilution precursor frequency studies showed the level of functional cells in the outer cortex was extremely low. The overall conclusion was that the vast majority of cells of complete "mature" phenotype are confined to the thymic medulla. These findings favor the view that thymus migrants originate from the thymic medulla, but do not exclude a cortical origin. The results also illustrate the need for multiparameter analysis to distinguish medullary thymocytes from early blast cells.     

Cytological identification of the lymphocytes in highly leukemic AKR/J mice in ontogenesis. Effect of somatotropin on the cellular composition of the lymphoid organs]

In AKR/J mice lymphocytes of thymus, spleen and lymph nodes can be classified according to their morphology as revealed by the Stockinger and Kellner staining: I--basophilic cytoplasm, 1--3 large basophilic nucleoli (blast cells), II--basophilic cytoplasm, 1--4 small clearly visible nucleoli (T-lymphocytes), III-weakly basophilic cytoplasm, 1-4 small fairly visible nucleoli (B-lymphocytes). The number of cells of types I and II increases with the progressing of leucaemia. After the administration of somatotrophic hormone the number of cells of types I and II increases only in the lymphoid organs of 5 months old AKR/J mice.     

CYTODYNAMICS IN THE THYMUS OF YOUNG ADULT MICE:

Cell proliferation and cell loss in the thymic blast cell population were studied in young adult mice by (1) stathmokinetic methods combined with an analysis of the PLMe-curve after a pulse ³H-TdR, and (2) nigrosin-dye exclusion combined with ³H-TdR-autoradiography. It was calculated that about 17% of the blast cells do not progress into mitosis within the period of an average cell cycle. The dye exclusion studies indicated a rate of blast cell death of about 2–5 %/hr. The two methods of assessing blast cell loss (death) support each other very well. In spite of these findings scintillation countings on thymuses removed from 1 to 17 hr after ³H-TdR injection showed fairly constant levels of thymic radioactivity. This suggests a very extensive reutilization of ³H-labelled break-down products from dying blast cells. The very sparse labelling of pyknotic thymocytes strongly suggests that thymic blast cells do not become pyknotic. The rate of small thymocyte production and disappearance was studied by pulse and repeated ³H-TdR labelling techniques combined with dye exclusion studies and pyknotic counts. The data from the repeated labelling experiment were analysed by use of a model based on the assumption of first order kinetics of small viable, dead, and pyknotic thymocytes. The rate of cell production was estimated to 1–6 %/hr whereas the rates of cell loss due to disintegration, i.e. supravital stainability and nuclear pyknosis, were calculated to 0–02 %/hr and 0–0006 %/hr respectively. Cell loss due to disintegration was less than 2 % of the total loss of small thymocytes. It was concluded that pyknotic counts are a useless method of assessing the cell death in the population of thymic blast cells and small thymocytes. On the basis of a model for thymocyte proliferation, production and loss it is suggested that about 45 % of the small viable thymocytes re-enter the generative cell pool, whereas about 55% disappear by emigration.     

A possible iridovirus in erythrocytes of Bufo marinus in Costa Rica

Icosahedral viral particles were found in the cytoplasm of erythrocytes and splenic reticular cells of a marine toad (Bufo marinus) collected from Costa Rica. Capsids had a maximum diameter of 312 nm and a spherical core with biphasic electron density. Viruses in erythrocytes were associated with cytoplasmic assembly areas and vacuoles in cytoplasm. Nuclei had finely granular material of decreased electron density located centrally, but contained no viral particles. A group of unenveloped viral particles was seen extracellularly in a splenic vessel. The virus was consistent with an iridovirus. In a blood smear stained with Giemsa round basophilic bodies with average diameters of 1.70 microns and morphologically similar to Pirhemocyton sp. were seen in the cytoplasm of erythrocytes and occasionally in the cytoplasm of monocytes or extracellularly. Erythrocytes containing these bodies had vacuoles and irregular pale-staining areas in the cytoplasm and pale-staining areas in the nucleus. These changes corresponded to the viral particles, assembly areas, vacuoles and nuclear changes at the ultrastructural level.     

Changes in the radiosensitivity of cortical thymocytes during differentiation]

To determine the radiosensitivity of cortical thymocytes at different stages of their differentiation, that took three days, estimated was the part of DNA of labelled thymocytes within the salt soluble fragments of the chromatin formed 6 h after irradiation of rats at different times during three days following the initial labelling of thymic blast DNA. The radiosensitivity of cells, which were at the terminal stages of differentiation, was twice as high as that of lymphocytes which were at the initial stage of differentiation.     

Immunocytochemical demonstration of glucocorticoid receptors in different cell types and their translocation from the cytoplasm to the cell nucleus in the presence of dexamethasone

Immunofluorescence microscopy has been applied to detect glucocorticoid receptors in rat thymocytes, HeLa cells and human mononuclear cells from peripheral blood. Blast formation induced in human mononuclear cells by PHA results in increased receptor concentration in the cytoplasm, as suggested from the immunofluorescence technique. Incubation of the blast cells with 10⁻⁷ M dexamethasone at 37°C within 15 min leads to decrease of staining in the cytoplasm and concomitant increase in the nucleus, indicative of a translocation of the cytoplasmic receptor into the nucleus.     

The stem cell antigens Sca-1 and Sca-2 subdivide thymic and peripheral T lymphocytes into unique subsets

Stem cell Ag 1 and 2 (Sca-1 and Sca-2), so named due to their expression by mouse bone marrow stem cells, were evaluated for expression by populations of cells within the thymus. Immunohistochemical analysis demonstrated that Sca-1 was expressed by cells in the thymic medulla and by some subcapsular blast cells, as well as by the thymic blood vessels and capsule. Sca-2 expression, which was limited to the thymic cortex, could be associated with large cycling thymic blast cells. Both Sca-1 and Sca-2 were expressed on a sub-population of CD4-CD8- thymocytes, and this subpopulation was entirely contained within the Ly-1lo progenitor fraction of cells. Sca-1 expression by a phenotypically mature subset of CD4+CD8- thymocytes was also noted. Conversely, Sca-2 expression was observed on a phenotypically immature or nonmature subpopulation of CD4-CD8- thymocytes. MEL-14, an antibody that defines functional expression of a lymphocyte homing molecule, identified a small population of thymocytes that contained all four major thymic subsets. Sca-2 split the MEL-14hi thymocyte subset into two Sca-2+ non-mature/immature phenotype fractions and two Sca-2- mature phenotype fractions. In peripheral lymphoid organs, Sca-1 identified a sub-population of mature T lymphocytes that is predominantly CD4+CD8-, in agreement with the thymic distribution of Sca-1. Peripheral T cells of the CD4-CD8+ phenotype were predominantly Sca-1-. In contrast, Sca-2 did not appear to stain peripheral T lymphocytes, but recognized only a subset of B lymphocytes which could be localized by immunohistochemistry to germinal centers. Thus, expression of Sca-1 is observed throughout T cell ontogeny, whereas Sca-2 is expressed by some subsets of thymocytes, including at least one half of thymic blasts, but not by mature peripheral T lymphocytes.     

STUDIES ON THE PERMEABILITY OF LYMPHATIC CAPILLARIES

The passageway for interstitial fluids and large molecules across the connective tissue lymph interface has been investigated in dermal lymphatic capillaries in the ears of guinea pigs. Numerous endothelial cells overlap extensively at their margins and lack adhesion devices at many points. The observations suggest that these sites are free to move as a result of slight pressure changes. Immediately following interstitial injections of tracer particles (ferritin, thorium, carbon, and latex spheres), many of the overlapped endothelial cells are separated and thus passageways are provided between the interstitium and lymphatic lumen. Tracer particles also occur in plasmalemmal invaginations along both connective tissue and luminal fronts. All of the tracer particles accumulate within large autophagic-like vacuoles. Very few particles of ferritin are observed in the endothelium after 24 hr; however, the vesicles containing the nonprotein tracer particles (carbon, thorium, and latex) increase in size and content and remain within the lymphatic endothelial cells up to 6 months. The role of vesicles in the transport of large molecules and particles is discussed in relation to the accretion of tracer particles within large vesicles and autophagic-like vacuoles in the endothelial cytoplasm.     

Ultrastructure of the crayfish kidney--coelomosac, labyrinth, nephridial canal

Regions of the crayfish kidney were examined by electron microscopy. Coelsmosac cells are loosely bound together by desmosome-like spot junctions, and connected to the basal lamina via characteristic pedicels. The cytoplasm contains numerous vesicles and vacuoles of various sizes and is often crowded with large, lysosome-like granules or dense bodies. The morphology suggests a filtration mechanism with reabsorption of materials such as protein from the filtrate and secretion of other substances into the lumen. The labyrinth is composed of cuboidal to columnar cells which possess a brush border, long and narrow intercellular spaces, basal plasmalemmal invaginations and typical cytoplasmic components. Two sub-regions are distinguishable. The morphology of labyrinth I suggests that these cells move fluid isotonically across the epithelium. Labyrinth II, in addition to isotonic transport, appears to be more active in the endocytic uptake and intracellular digestion of large molecules such as protein. The nephridial canal consists of cells which lack a brush border, but display extensive basal invaginations associated with elongated mitochondria. A proximal and distal region are cytologically distinguishable. Proximally, the cells are small and filled with mitochondria throughout. Scattered within the cytoplasm are vesicles, vacuoles, diffuse glycogen, free ribosomes, dense bodies and some rough endoplasmic reticulum. Distally, the cells are less compact, larger, and cuboidal to columnar in shape. The cytoplasm is similar to that of the proximal cells, but the basal invaginations are even larger and more extensive. The morphology of cells in both regions of the nephridial canal is highly suggestive of active solute reabsorption, probably occurring against an osmotic gradient.     

Fine Structures Associated with Nutrition of the Intracellular Parasite Eimeria auburnensis*

SYNOPSIS. In the nearly mature macrogametes of Eimeria auburnensis, the cell membrane is a unit membrane, with underlying and overlying osmiophilic layers usually present. Cup-shaped micropores were occasionally seen. Smaller, V-shaped invaginations were also found in considerable numbers at the surface. At the deepest point, these invaginations were bounded only by a unit membrane. Immediately adjacent to this point, vesicles with homogenous electron-pale contents bounded by a similar unit membrane, were frequently seen. Pinocytosis evidently occurs at the site of these invaginations. Numerous folds of the host cell membrane bordering the vacuole in which the parasite lay extended about 0.1–0.7 μ into the vacuole. These “intravacuolar folds” varied in depth and number in different specimens. In some, the majority of folds had apparently become disconnected from the host cell membrane. A highly developed smooth endoplasmic reticulum occurred in the adjacent host cell cytoplasm. The intravacuolar folds may assist in transfer of nutrients, including membrane material, from the host cell to the parasite. The evidence indicates that in this species of Eimeria nutrients are taken into the parasite primarily as fluids by pinocytosis and possibly other processes.