bluuebunny/crossref_metadata_2025 huggingface dataset:
THE IN VIVO REUTILIZATION OF LYMPHOCYTIC AND SARCOMA DNA BY CELLS GROWING IN THE PERITONEAL CAVITY
Authors: Rieke William O.
Published: May 2004

An ascites tumor, Sarcoma I, was transplanted to isologous and homologous mice which had been labeled with tritiated thymidine from 1 to 24 hours previously. Radioautographic preparations revealed labeled host lymphocytes emerging to mingle with the transplanted tumor and the subsequent appearance of nuclear radioactivity in the sarcoma. Sarcoma cells cultured subcutaneously or in Millipore diffusion chambers in previously labeled mice did not demonstrate significant radioactivity. Transplantation of washed, H3-thymidine-labeled lymphocytes to non-radioactive, sarcoma-bearing mice was followed by the gradual appearance of nuclear radioactivity in the sarcoma. The label in the sarcoma was entirely removed by deoxyribonuclease but not by ribonuclease treatment prior to radioautography. Intraperitoneal injections of purified, H3-thymidine-labeled sarcoma or lymphoid DNA in normal or tumor-bearing mice were followed by radioactivity appearing in sarcoma or normal peritoneal mononuclear cells. It was concluded that reutilization of DNA and its metabolites may occur in vivo, and the conditions under which reutilization may be detected are discussed.
SPECTROPHOTOMETRY OF THE PERIODIC ACID-SCHIFF REACTION WITH PITUITARY HORMONES IN VITRO AND IN HISTOLOGICAL SECTIONS
Authors: Fand Sally B. , Thorell Bo
Published: May 2004

The spectral light absorption of the in vitro periodic acid-Schiff reactions of 4 purified pituitary hormones is described. The absorption spectra present a maximum between 560 and 565 mµ. The color developed conforms with Beer's law for the ranges of concentration examined. The different hormones exhibit different chromogenicity per unit of biological activity: the color produced by 1 unit of FSH is equivalent to approximately 2 of TSH, 4 of LH, and 30 of ACTH. Microspectrophotometric measurements of the PAS-positive structures in histological sections of the human pituitary give absorption curves with shapes similar to those obtained in vitro, although quantitative differences exist. It is concluded that under the proper experimental conditions microspectra of the pituitary structures might, in the future, prove to give a quantitative measure of aldehyde groups generated from glycoprotein tropins by periodate oxidation.
THE SPECIFIC VOLUME OF DRY PROTEIN
Authors: Brill A. S. , Olin J. S. , Siegel B. M.
Published: May 2004

The specific volume of dry protein is of interest in the interpretation of electron micrographs of protein molecules. A pycnometer with helium gas as the fluid is described and measurements on dry powders of lysozyme, ovalbumin, bovine serum albumin, and fibrinogen are reported. The values obtained exceed those from determinations on solutions by an average of 1.4 per cent when the latter measurements are uncorrected for electrostriction.
THE EFFECT OF A MEDIUM DOSE (430 ROENTGENS) OF X-RAY IRRADIATION ON RESTING CELLS OF THE LIVER
Authors: Horst Antoni , Rudnicki Tadeusz
Published: May 2004

The mitotic activity of regenerating liver cells after a single dose (430 r) of x-ray irradiation was studied. In every group of the experimental animals (white rats), the mitotic activity (mitotic index) and the number of abnormal mitotic figures were determined. The results indicated that resting cells irradiated a short time before mitotic activity showed reactions similar to those of cells irradiated during mitotic activity. The disturbances in the irradiated mitotically active cells were only quantitatively different from those in the irradiated resting cells. The disturbances in the irradiated resting cells depended upon the time interval between the irradiation and the beginning of mitotic activity stimulated by partial hepatectomy. It was found that the shorter the time interval, the more pronounced were the disturbances and the more similar they became to those of irradiated mitotically active cells. Conversely, the longer the time interval between the irradiation and the beginning of mitotic activity, the less pronounced were the disturbances and the more similar they became to those of the non-irradiated control cells. A discussion is presented as to whether or not the lesions of resting cells caused by a single medium dose of x-ray irradiation are reversible, and whether such lesions are only brought to light by the process of mitosis or whether the process of mitosis renders it possible for these lesions to develop.
OPTICAL AND ELECTRON MICROSCOPIC CHANGES IN ULTRAVIOLET-IRRADIATED CHROMOSOME SEGMENTS
Authors: Bloom William , Leider Robert J.
Published: May 2004

Chromosome segments of urodele cells lose some substance after irradiation with about 10-1 ergs/µ2 of heterochromatic ultraviolet light. These segments stain faintly or negatively with the Feulgen and pyronine-methyl-green methods and weakly with the Alfert-Gesch-wind stain for basic protein. In the living cells, Perry found in these chromosome segments a decrease of 50 to 60 per cent in absorption at 2400, 2600, and 2800 A, i.e., in the region of intense chromosomal absorption that is maximal at 2600 A. Apparently the material lost contains DNA (?DNP) and we call the process DNA-steresis. In such cells, fixed in neutral formalin in Tyrode's solution and stained with phosphotungstic acid, electron microscopy shows that the unirradiated parts of the chromosomes consist of (a) a homogeneous or finely fibrillar material (component-A) filling the meshes of (b) an irregular network with bars 40 to 300 A in diameter, some of which continue into a similar interchromosomal network. DNA-steretic portions of the chromosomes consist mainly of this network and only small amounts of component-A, which presumably contains the DNA. We have not been able to demonstrate DNA-steresis with the electron microscope after primary fixation with OsO4 or KMnO4. Structural changes due to DNA-steresis are compared with certain nuclear changes in the mitotic cycle.
AN ELECTRON MICROSCOPE STUDY OF THE EARLY ASSOCIATION BETWEEN TWO MAMMALIAN VIRUSES AND THEIR HOSTS
Authors: Dales Samuel
Published: May 2004

Early interaction between two animal viruses, vaccinia and adenovirus 7, which multiply readily in L strain and HeLa cells, respectively, was examined in both whole mount preparations and in thin sections. To observe the association at the surface, cells carrying adsorbed virus were swelled under controlled conditions and then "stained" with neutral phosphotungstate. Each particle of both virus types becomes attached to the cell by several capsomeres and is then ingested by phagocytosis. Within the cell, near the surface, single particles or small clumps of adenovirus are lodged within vesicles. Deeper in the cytoplasm this virus is packed in large, numerous inclusions, whereas very close to the nuclear envelope only free particles are found. Vaccinia, on the other hand, either free or in vesicles, is always found in the cytoplasm, at some distance from the nucleus (11). Adsorption and intracellular disposition of these two viruses is discussed in relation to the infectious process.
THE ULTRASTRUCTURE OF THE Z DISC IN SKELETAL MUSCLE
Authors: Knappeis G. G. , Carlsen F.
Published: May 2004

This electron microscopic study deals with the structure of the Z disc of frog's skeletal muscle, with special regard to the I filaments—whether they pass through the Z disc or terminate at it. In most longitudinal sections the I filaments terminate as rod-like projections on either side of the Z disc, one I filament on one side lying between two I filaments on the opposite side. This indicates that the I filaments are not continuous through the Z disc. The rod-like projections are often seen to consist of filaments (denoted as Z filaments) which meet at an angle.

In cross-sections through the Z region the I filaments and Z filaments form tetragonal patterns. The I filaments are situated in the corners of the squares; the oblique Z filaments form the sides of squares. The tetragonal pattern formed by the Z filaments is rotated 45 degrees with respect to the tetragons formed by the I filaments on both sides of Z. This structural arrangement is interpreted to indicate that each I filament on one side of the Z disc faces the center of the space between four I filaments on the opposite side of Z and that the interconnection is formed by four Z filaments.
SYNAPTIC STRUCTURES IN THE LATERAL LINE CANAL ORGAN OF THE TELEOST FISH LOTA VULGARIS
Authors: Flock Åke , Wersäll Jan
Published: May 2004

This paper is a preliminary report on some of the results of electron microscopic studies on the lateral line canal organ of the teleost fish Lota vulgaris. It deals with the ultrastructure of the synaptic area on the hair cells of the sensory epithelium and describes the nerve endings as well as a complicated system of foldings of the hair cell plasma membranes enclosing portions of the hair cell cytoplasm in the synaptic area. These findings are discussed in the light of present knowledge of the ultrastructure of other receptoneuronal synapses.
INTRACELLULAR SYNTHESIS OF CHONDROITIN SULFATE
Authors: Dziewiatkowski Dominic D.
Published: May 2004

In autoradiograms of slices of costal cartilage, incubated for 4 hours in a salt solution containing S35-sulfate and then washed extensively and dehydrated, about 85 per cent of the radioactivity was assignable to the chondrocytes. From alkaline extracts of similarly prepared slices of cartilage, 64 to 83 per cent of the total sulfur-35 in the slices was isolated as chondroitin sulfate by chromatography on an anion-exchange resin. In view of the estimate that only about 15 per cent of the radioactivity was in the matrix, the isolation of 64 to 83 per cent of the total sulfur-35 as chondroitin sulfate is a strong argument that the chondrocytes are the loci in which chondroitin sulfate(s) is synthesized.
SUBSURFACE CISTERNS AND THEIR RELATIONSHIP TO THE NEURONAL PLASMA MEMBRANE
Authors: Rosenbluth Jack
Published: May 2004

Subsurface cisterns (SSC's) are large, flattened, membrane-limited vesicles which are very closely apposed to the inner aspect of the plasma membranes of nerve cell bodies and the proximal parts of their processes. They occur in a variety of vertebrate and invertebrate neurons of both the peripheral and central nervous systems, but not in the surrounding supporting cells. SSC's are sheet-like in configuration, having a luminal depth which may be less than 100 A and a breadth which may be as much as several microns. They are separated from the plasmalemma by a light zone of ∼50 to 80 A which sometimes contains a faint intermediate line. Flattened, agranular cisterns resembling SSC's, but structurally distinct from both typical granular endoplasmic reticulum (ER) and from Golgi membranes, also occur deep in the cytoplasm of neurons. It is suggested that membranes which are closely apposed may interact, resulting in alterations in their respective properties. The patches of neuronal plasmalemma associated with subsurface cisterns may, therefore, have special properties because of this association, resulting in a non-uniform neuronal surface. The possible significance of SSC's in relation to neuronal electrophysiology and metabolism is discussed.
ELECTRON MICROSCOPY OF SPORES OF BACILLUS MEGATERIUM WITH SPECIAL REFERENCE TO THE EFFECTS OF FIXATION AND THIN SECTIONING
Authors: Rode L. J. , Lewis C. Willard , Foster J. W.
Published: May 2004

Resting spores of Bacillus megaterium appear uniformly opaque and undifferentiated under the electron microscope. Germinated spores and spores which have lost their dipicolinic acid underwent characteristic changes in structure. Spores fixed with KMnO4 lose their dipicolinic acid. Spores fixed with OsO4 under certain conditions retain their dipicolinic acid. When conventional sectioning procedures are used with either method of fixation, abnormal spore structure is produced as a result of the solution of cellular constitutents. Dry sections of unfixed spores embedded in methacrylate reveal the spore structure in a more normal state. Indirect evidence has been obtained for the existence of a penetration barrier at or near the outer edge of the cortex.
FORMATION AND STRUCTURE OF THE SPORE OF BACILLUS COAGULANS
Authors: Ohye D. F. , Murrell W. G.
Published: May 2004

Spore formation in Bacillus coagulans has been studied by electron microscopy using an epoxy resin (Araldite) embedding technique. The developmental stages from the origin of the initial spore septum to the mature spore were investigated. The two forespore membranes developed from the double layer of cytoplasmic membrane. The cortex was progressively deposited between these two membranes. The inner membrane finally became the spore protoplasmic membrane, and the outer membrane part of the inner spore coat or the outer spore coat itself. In the mature spore the completed integuments around the spore protoplasm consisted of the cortex, a laminated inner coat, and a dense outer coat. No exosporium was observed. The method of formation of the cortex and the spore coats is discussed.
AN ELECTRON MICROSCOPE STUDY OF RADIATION DAMAGE IN THE MOUSE OOCYTE
Authors: Parsons Donald F.
Published: May 2004

Cytological changes occurring in young oocytes of the mouse, following whole-body x-irradiation, have been examined by light and electron microscopy. Two minutes after a dose of 200 r, a drop occurs in the number of mitochondria per oocyte. The normal number of mitochondria is restored in the next few minutes. Five to 6 hours after a dose of 7 r, and 30 to 45 minutes following a dose of 200 r, all the oocytes are markedly contracted. In the next 2 hours (at a dose of 7 r), some cells shrink further and become pycnotic, while others expand and show signs of karyolysis. Of the expanded cells, some (about 50 per cent of total young oocytes) become morphologically normal. In both pycnotic and karyolytic nuclei, the nucleolus was contracted but without loss of its fibrillar structure. The contracted nucleolus appeared similar to those seen in some nearly mature normal oocytes (these oocytes have a high natural death rate). No other cell type in the ovary was affected by doses of x-rays up to 200 r. Observations on the nucleus of the normal oocyte are included. In the dictyate stage of meiotic prophase the chromosomes were dispersed into bundles of 100 A microfibrils. The main component of the nucleolus was found to be tightly coiled fibrils of 60 to 100 A, which appear to have a close relationship with the microfibrils of the chromosomes.
INCORPORATION OF TRITIATED CYTIDINE INTO RIBONUCLEIC ACID BY ISOLATED PEA NUCLEI
Authors: Rho Joon H. , Chipchase Margaret I.
Published: May 2004

A new method for the isolation of the nuclei from plant tissue has been devised. This method consists in passing the plant tissue through a set of spring-loaded, counter-rotating rollers and collecting the liberated nuclei in sucrose solution containing calcium ions. A semi-automatic machine which couples the rollers with a special tissue chopping device has been constructed. It has been shown that nuclei obtained in this way actively incorporate cytidine-H3 into RNA. This incorporation is increased in the presence of nucleoside triphosphates and an energy-regenerating system.
UNUSUAL FORMATIONS OF ERGASTOPLASM IN PAROTID ACINOUS CELLS OF MICE
Authors: Parks Harold F.
Published: May 2004

The ergastoplasm (granular endoplasmic reticulum) of parotid acinous cells of the mouse is described with special emphasis on unusual forms of this membranous system. In the majority of the acinous cells the ergastoplasm appeared in sections to consist of a large number of separate flattened cisternae. In some acinous cells, however, the ergastoplasm was disposed as a very small number of large membranous formations. Although extensive and complicated in form, these latter formations could be seen, from the examination of a single section, to consist of a single expanse of membrane continuous with the nuclear envelope. Certain acinous cells exhibited ergastoplasmic formations which appeared to represent intermediate stages of a fragmentation or metamorphosis of the larger formations toward the more usual form of ergastoplasm. These observations suggest the possibility that the early elaboration of ergastoplasm consists in the production, in relation to the nuclear envelope, of large, complicated membranous formations that subsequently sever their connection with the nuclear envelope and form a large number of separate, or tenuously connected, cisternae. The majority of the large, complicated ergastoplasmic formations were seen in parotid glands of mice that had been starved for 4 days and subsequently fed for a variable number of hours, but some were found in glands that were not subjected to experimental treatment. The tissues studied were prepared for electron microscopic examination by fixation in osmium tetroxide, dehydration in alcohol, imbedding in butyl methacrylate, sectioning with a glass knife, staining with lead hydroxide, and sandwiching with formvar.
STUDIES ON PROTEIN UPTAKE BY ISOLATED TUMOR CELLS
Authors: Ryser H. , Caulfield J. B. , Aub J. C.
Published: May 2004

Ferritin, added to the incubation medium of ascites tumor cells, was used as an electron microscopic marker to study the uptake of large protein molecules by morphologically intact cells. A definite uptake could be detected after 1 hour of incubation in Tyrode bicarbonate solution containing 0.04 to 13.3 mg ferritin/ml. Ferritin was found in a variety of membrane-surrounded structures, suggesting that pinocytesis and related membrane movements are occurring under physiological conditions and can account for the penetration of intact macromolecules into isolated tumor cells. Supplementation of the medium with serum albumin (33 mg/ml) increased the average amount of ferritin per cell and per pinocytotic structure. Ferritin was strongly adsorbed by fragments of lysed cells, which were readily taken up by intact cells. Besides its role as carrier, this debris appeared to stimulate membrane movements. Only rare examples were found to suggest the release of ferritin from the pinocytotic structures into the cytoplasm. Thus, the disintegration of such structures cannot be considered an obvious step towards a rapid metabolic utilization of protein by the cell. Particles of colloidal gold presented to the cell under the same conditions were not taken up to any significant extent, thus providing good evidence for a selective ingestion of particles of comparable sizes.
A COMPARISON OF THE CHANGES IN FINE STRUCTURE OF L CELLS DURING SINGLE CYCLES OF VIRAL MULTIPLICATION, FOLLOWING THEIR INFECTION WITH THE VIRUSES OF MENGO AND ENCEPHALOMYOCARDITIS
Authors: Dales Samuel , Franklin Richard M.
Published: May 2004

The virus of encephalomyocarditis (EMC), examined by the negative-contrast method, is indistinguishable from the serologically related Mengovirus. The particles are 270 to 280 A in diameter. The surface of EMC is composed of an undetermined number of subunits. Frequent sampling of infected cells was carried out throughout one-step cycles of viral multiplication to observe cytopathic changes occurring in L cells infected by these two related RNA viruses. EMC and Mengovirus, which multiply at equal rates, in most respects elicit similar alterations in cell fine structure. Rearrangement and changes in nuclear material accompanied by formation of small vesicles in the centrosphere region commence at 4 to 6 hours after infection. Thereafter a progressive degeneration of the nucleus and vesiculation of the cytoplasm are observed up to 18 to 20 hours. Increased numbers of small dense granules, indistinguishable from ribonucleoprotein particles, appear in the cytoplasm between 8 and 14 hours after infection. L cells infected with Mengovirus become permeable to Erythrocin more slowly than those infected with EMC. Only in the case of Mengovirus infection are large aggregates of dense material first observed in the cytoplasm at 8 hours, followed by the appearance of crystals probably composed of Mengovirus particles, at 12 hours. Differences in the rates of cell permeability after infection with EMC and Mengovirus are discussed in relation to formation of virus crystals and plaque-type mutants.
THE ORIGIN OF PROTEIN AND FATTY YOLK IN RANA PIPIENS
Authors: Ward Robert T.
Published: May 2004

Electron microscope studies of young oocytes have demonstrated that the plate-like, hexagonally shaped yolk bodies previously observed in living cells are wholly within the substance of oocyte mitochondria and that they remain within these mitochondria while increasing in size. These bodies possess a crystalline structure consisting of what appear to be lines, with a spacing of 70 to 85 A, and appear very dense in the electron microscope. After formalin fixation such bodies give an intense positive test for protein, and when viewed in the electron microscope are only slightly less dense than after OsO4 fixation. Evidence is presented for the origin of these crystals within a single crista. The clusters of yolk globules previously studied in living cells are seen to consist of several types of bodies, but an irregular dense droplet predominates. This dense material is apparently secreted by small spherical bodies which, the evidence suggests, originate from the breaking up of filamentous mitochondria and which possess an outer double membrane and sometimes internal cristalike membranes. When thin sections of young oocytes are immersed in xylol the dense globules of the clusters are dissolved, but the hexagonal bodies are unaffected, indicating that the globules are of a predominantly fatty nature, while the hexagonal bodies are of a predominantly protein nature. Examination of mature or almost mature oocytes has revealed that the main body of the yolk platelets is crystalline in nature and is surrounded by a thick matrix which, in light microscope study, masks the fact that the face view of the main body of the platelets is often hexagonal. The spacing within the main body is found to be 70 to 85 A. The crystal laminae of this material can be resolved quite clearly into rows of particles. Dense globules of varying sizes are found in the cytoplasm between the platelets. When thin sections of these OsO4-fixed oocytes are immersed in xylol, the material of the globules is extracted and the crystalline material of the platelets remains unaffected, indicating the fatty nature of the globules and the protein nature of the platelets. The platelets of the mature egg resemble the hexagon bodies, previously described in young oocytes, in their protein nature, their crystalline spacing, and their hexagonal outline. This is given as strong evidence for the origin of the mature platelets by the growth of the intramitochondrial hexagon bodies. The biochemical implications of this study are discussed.
THE PERIOD OF DNA SYNTHESIS PRIOR TO MEIOSIS IN THE MOUSE
Authors: Lima-de-Faria A. , Borum Kirstine
Published: May 2004

Sixteen pregnant female mice were operated on and H3-thymidine was injected into the amniotic cavities of the uterus. The injection was given between the 6th and 14th days of fetal life. Eighty-eight fetuses received thymidine in this way. Another series of 16 pregnant females was injected intraperitoneally with H3-thymidine between the 5th and 14th days of pregnancy. Two of these females were killed 16 days after the observation of the vaginal plug. The remaining 30 females were allowed to give birth to their progeny. The progeny was killed at birth and the ovaries of the newborn females fixed at once. Labeled oocytes at late pachytene and early diplotene were clearly seen in individuals that received the isotope between the 10th and 12th to 13th days of fetal life, but the period of DNA synthesis preceding meiosis is at the 12th to 13th days of fetal life. Since meiosis is recognized by the 14th day, only the oocyte labeling originating from mothers injected at the 12th and 13th days may be considered as representing the DNA synthesis of the premeiotic replication.
ADENOSINE TRIPHOSPHATE-LINKED CONCENTRATION OF CALCIUM IONS IN A PARTICULATE FRACTION OF RABBIT MUSCLE
Authors: Ebashi Setsuro , Lipmann Fritz
Published: May 2004

ATPase and ATP-dependent calcium ion concentration was studied with a membrane fraction isolated from homogenized rabbit skeletal muscle by differential centrifugation. Electron micrographs of the fraction indicate that it consists mainly of resealed tubules and vesicles of the endoplasmic reticulum. The up-to-1400-fold concentration of calcium in this fraction might be explained by proposing the existence of an energy-requiring system for the transport of calcium ions into the tubules or vesicles.
CELLULAR INJURY IN VITRO: PHASE CONTRAST STUDIES ON INJURED CYTOPLASM
Authors: Buckley I. K.
Published: May 2004

Unfixed, compressed acinar cells of rat pancreas, isolated by mechanical and enzymatic means, were examined by phase microscopy and photomicrographed using 35 mm film and electronic flash illumination. Similarly, observations were made on Walker carcinoma cells; in addition, these cells were treated with solutions containing either phosphatidase A or enzyme inhibitors. Acinar cells contained, besides nuclei, perinuclear droplets and secretion granules, various membranous and vacuolar structures. The basal cytoplasm showed parallel dark lines interpreted as endoplasmic reticulum. In some cells, fragmentation of the reticulum was followed by the direct incorporation of fragments into simple myelin figures. In other cells it appeared that phase-lucent linear structures and vacuoles were derived by dilatation of cisternae of the endoplasmic reticulum. Perinuclear fluid collections arose either by dilation of the perinuclear cisternae of the endoplasmic reticulum or by fluid dilatation of the nuclear envelope. Phosphatidase A disrupted early vacuoles of Walker carcinoma cells. From this and the direct involvement of elements of the endoplasmic reticulum in myelin figures, it was concluded that the membranes limiting the endoplasmic reticulum incorporate phosphatides in continuous layers. While many severely injured cells formed large vacuoles, others developed concentrically laminated myelin figures; it was concluded that both types of structure derived from phosphatides liberated intracellularly, the vacuoles by vesicular myelin figure formation.
NUCLEAR ENVELOPE-CHLOROPLAST RELATIONSHIPS IN ALGAE
Authors: Gibbs Sarah P.
Published: May 2004

In Ochromonas danica and two related species (Chrysophyceae) and in Rhodomonas lens and Cryptomonas sp. (Cryptophyceae), the chloroplast is surrounded by an outer double-membraned envelope which lies outside the usual double-membraned chloroplast envelope. At the borders of the area where the chloroplast lies adjacent to the nucleus, this outer envelope is continuous with the outer membrane of the nuclear envelope as a double-membraned outfolding, so that the entire chloroplast in these species lies within a double-membraned sac, one wall of which is the nuclear envelope. In Olisthodiscus sp. (Chrysophyceae ?), each of the small peripheral chloroplasts is surrounded by a similar double-membraned outer envelope, but in this species no connections with the nuclear envelope were observed. In the Ochromonadaceae, a characteristic array of tubules is present within the sac in the narrow space which separates the chloroplast from the nucleus. In the other species studied, tubules are present at places between the chloroplast envelope and the outer envelope. In the Cryptophyceae, the starch grains lie outside the chloroplast envelope, but within the outer double-membraned sac. A double-membraned outer envelope appears to be present outside the chloroplasts of the Phaeophyta and Euglenophyta, but seems to be absent in the other groups of algae.
THE FINE STRUCTURE OF THE PIGMENT EPITHELIUM IN THE ALBINO RAT
Authors: Dowling John E. , Gibbons I. R.
Published: May 2004

In this report, particular attention is paid to the inclusion bodies found in the apical cytoplasm of the pigment epithelial cell. These bodies are of variable size and form. The smallest (0.4 µ diameter) consist of a granular matrix enclosed by a single membrane, and are similar to the lysosomes of hepatic cells. Larger inclusion bodies contain areas of lamellated material in addition to granular matrix. The largest particles seen (2 µ diameter) are almost entirely lamellar. These different forms seem closely related, for it is possible to find all transitional stages between the smallest and largest particles. The relationship between the lamellar inclusion bodies and the rod outer segments is discussed.
THE ISOLATION OF CELL NUCLEI FROM RAT BRAIN
Authors: Sporn Michael B. , Wanko Theodor , Dingman Wesley
Published: May 2004

A method for preparing highly pure cell nuclei from adult rat brain, using both differential and isopycnic centrifugation in sucrose media, is described. The morphology of these preparations was examined by both phase contrast and electron microscopy. The isolated nuclei retained many aspects of their in situ morphology; in particular, the nuclear envelope was double layered and interrupted by pore-like discontinuities, and the nucleoli consisted of irregular masses of densely packed granules. Analyses of these nuclear preparations for cytochrome oxidase and cholinesterase activity, as well as RNA/DNA ratio, indicated minimal contamination with mitochondria and microsomes. Problems involving the homogenization technique, choice of ionic conditions in the homogenization medium, and choice of optimal density of the sucrose solution used for the final purification of nuclei are discussed. Results of application of the technique to isolation of adult rat liver nuclei are also reported.
A STUDY OF THE ORIENTATION OF THE SENSORY HAIRS OF THE RECEPTOR CELLS IN THE LATERAL LINE ORGAN OF FISH, WITH SPECIAL REFERENCE TO THE FUNCTION OF THE RECEPTORS
Authors: Flock Åke , Wersäll Jan
Published: May 2004

The morphology of the hair bundles on top of the receptor cells in the lateral line organ of the teleost fish Lota vulgaris is described. Each receptor cell shows a distinct morphological polarization. Two groups of receptor cells can be distingiushed, one consisting of cells polarized towards the head, the other consisting of cells polarized towards the tail. In the crista ampullaris all cells are polarized in the same direction. An hypothesis is proposed for the function of the receptor cells in the lateral line organ and the labyrinth based on a correlation of morphological and functional polarization.
FURTHER STUDIES ON THE THETA CELL OF THE MOUSE ANTERIOR PITUITARY AS REVEALED BY ELECTRON MICROSCOPY, WITH SPECIAL REFERENCE TO THE MODE OF SECRETION
Authors: Sano Masao
Published: May 2004

Theta cells reported previously as a new cell type in the anterior pituitary of the mouse were examined with the electron microscope. This type of cell is distinguished by the presence of pleomorphic secretory granules, a characteristic arrangement of the rough surfaced variety of endoplasmic reticulum, a well developed Golgi complex, and an eccentrically located nucleus. The secretory granules are seen at first as small granules of low density within the Golgi vesicles. While they are within the Golgi vesicles they become larger and denser. Simultaneously they move from the proximal to the distal part of the Golgi region and finally emerge from the Golgi area as mature granules in the cytoplasm. Thus, secretory granules are always enveloped by a limiting membrane which originates from the wall of the Golgi vesicle. At the stage of granule-extrusion, the cell membrane fuses with the limiting membrane of the granules and openings in the cell membrane appear at the place of extrusion. The granules then appear to lie within inpocketings of the cell membrane. They lose their density within these inpocketings or within the cytoplasm and occasionally show fragmentation. After complete loss of density, the granules are extruded as amorphous materials to the territory outside of the cell.
WOUND HEALING AND COLLAGEN FORMATION
Authors: Ross Russell , Benditt Earl P.
Published: May 2004

The sequence of incorporation and utilization of tritium-labeled proline has been examined in healing wounds from normal and scorbutic guinea pigs. Linear incisions in the skin of the animals were allowed to heal for 7 days. Each animal was given proline-H3, and the wounds were excised 30 minutes, 1 and 4 hours, 1, 3 and 7 days after proline administration. The tissues were fixed in osmium tetroxide, fixed again in neutral buffered formalin, embedded in epoxy resin, and sectioned at 1 micron thickness. The sections were coated with nuclear track emulsion, exposed, developed, and stained. The results of grain counts were quantitated as the number of counts per unit area overlying cells, fibers, etc. In both groups the proline reaches a maximum over the fibroblasts within 4 hours and subsequently disappears from the cells. Concomitantly, the proline reaches a maximum over the collagen (in normal animals) and extracellular fibrillar material (in scorbutic animals) by 4 hours, where it remains. The modified technique of radioautography used in this study allows not only resolution of approximately 1 micron, but also minimal background, decreased artifact, and a clear separation of the randomly situated elements within the wounds so that grain counting is facilitated. The results correlated with previous electron microscopic studies are consistent with the utilization of proline by the fibroblasts and its incorporation into collagen (in normal animals) and into the extracellular, fibrillar, non-collagenous material seen in scorbutic animals.
HIGH-RESOLUTION AUTORADIOGAPHY
Authors: Caro Lucien G.
Published: May 2004

The resolution obtainable in electron microscopic autoradiographs, using a photographic emulsion consisting of a monolayer of silver bromide crystals, was investigated theoretically and experimentally. The expected distribution of exposed crystals around a point source was calculated from the geometry of the preparation and from the range distribution of the beta particles emitted by tritium. From such a distribution an autoradiographic resolution of the order of 1000 A can be predicted. From the point source distribution, the expected distribution of grains around bacteriophages labeled with tritium was calculated. This distribution was also measured experimentally in electron microscopic autoradiographs of bacteriophages T-2 labeled with thymidine-H3. The two distributions agreed closely. It was also verified, using the nuclear region in thin cross-sections of Bacillus subtilis labeled with thymidine-H3, that resolutions of the same order were obtained for extended sources. It was concluded that an autoradiographic resolution of 1000 A could be achieved with a presently available commercial emulsion, although emulsions with finer grains might be desirable in some circumstances.
THE IDENTIFICATION OF CHYLOMICRA AND LIPOPROTEINS IN TISSUE SECTIONS AND THEIR PASSAGE INTO JEJUNAL LACTEALS
Authors: Casley-Smith J. R.
Published: May 2004

The electron microscopic appearances of chylomicra and lipoproteins have been investigated. The particles were isolated from rat chyle by differential flotation in an ultracentrifuge. Various fixing and embedding media were used. The two kinds of particles were then identified in thin sections of the jejunum of rats. The chylomicra had diameters of from 1,000 A to 1 µ; the lipoproteins ranged from 100 to 1,000 A. They were identified by their sizes and their similarities to the isolated particles after the various fixing and embedding procedures. In addition, the relative amounts of the two kinds of particle varied greatly under different dietary conditions. The chylomicra had a thin rim, probably of phospholipid. Section B records the passage of the two kinds of particle into the lacteals in the villi of the jejunum. Both chylomicra and lipoproteins were seen passing through many open junctions. From permeability considerations it would seem that this is the most important route. These open junctions appear to act as "inlet valves" which prevent backflow as the contractions of the villi pump material out of the lacteals. Both chylomicra and lipoproteins were also seen entering the endothelial cells and lying inside them. The lipoproteins entered via "normal" caveolae and were seen in "normal" vesicles (∼500 A); the chylomicra necessarily occupied much larger organelles. Both kinds of particles were also seen in caveolae on the luminal surface of the endothelium, but it was impossible to be certain that these were not just particles entering the cells from the lumen. The chylomicra often seemed to be washed out of these caveolae as many large, empty ones were seen on the luminal sides of the cells. Frequently, these caveolae had dark membranes.
THE FINE STRUCTURE OF THE BASEMENT MEMBRANE IN EPIDERMAL TUMORS
Authors: Frei J. V.
Published: May 2004

Epidermal tumors were induced in Swiss female mice by a topical application of 9,10-dimethyl-1,2-benzanthracene solution followed by repeated applications of croton oil solutions. Fourteen benign and malignant tumors were sampled 25 weeks after the treatment had begun, fixed in osmium tetroxide or potassium permanganate, and embedded in Epon. Sections stained with lead hydroxide were examined. In three tumors defects of the epidermal basement membrane were seen. These defects accompanied local invasion of the tumors. The possible mechanisms of the development of this unusual anatomical situation are discussed.
THE FINE STRUCTURE OF NORMAL AND NEOPLASTIC MELANOCYTES IN THE SYRIAN HAMSTER, WITH PARTICULAR REFERENCE TO CARCINOGEN-INDUCED MELANOTIC TUMORS
Authors: Rappaport Henry , Nakai Takashi , Swift Hewson
Published: May 2004

The dermal melanocyte system of the Syrian hamster is particularly responsive to the melanogenetic and tumor-inducing effects of 7,12-dimethylbenz(a)anthracene (DMBA). The melanocytes of the hair follicles appear to be susceptible to the melanogenetic effect of DMBA but not to its tumor-inducing effect. The epidermal melanocytes are non-pigmented and are unresponsive to both melanogenetic and carcinogenic effects of DMBA. The pigmented granules of the dermal melanocytes of both the golden and the white hamster have an identical substructure and pattern of melanization which occurs in an orderly fashion on a delicate fibrillar component. The hair melanocytes have larger pigment granules with a more complicated fibrillar substructure. The epidermal melanocytes do not possess pigment granules but are recognized by their dendritic shape, the absence of desmosomes and tonofilaments, and the presence of racket-shaped or rod-shaped organelles. The melanin granules in neoplastic melanocytes of the golden hamster differ from corresponding normal melanocytes only in their larger size. In the white hamster, however, the melanin granules in tumors produced under identical experimental conditions are so bizarre and atypical that consideration was given to the possibility that a genetic difference in the melanization pattern between the two varieties becomes apparent in carcinogen-induced melanotic tumors. No definite conclusions could be reached as to the precise origin of the melanin granules in either normal or neoplastic melanocytes.
OBSERVATIONS ON MITOCHONDRIAL STRUCTURE
Authors: Revel J. P. , Fawcett D. W. , Philpott C. W.
Published: May 2004

This paper reports the common occurrence of creases or sharp ridges on the membranes of the mitochondrial cristae. In section these appear as sharp angulations recurring at more or less regular intervals along the profile of the crista. In instances where such angulations occur alternately on one membrane and then on the other, the crista has a zig-zag course. Where they occur at the end of a crista its profile has a square tip. An exaggerated expression of this tendency for angulation of the internal mitochondrial membranes is found in certain bat muscles where some of the cristae take the form of parallel bundles of prismatic tubules which are triangular in cross-section. Angular configurations of the cristae have been observed after various methods of specimen preparation, in a variety of cell types, and in a wide range of animal species. They are believed to be a normal variation of the basic structural organization of the mitochondrion and to be the morphological expression of a property which the internal mitochondrial membranes do not share with other membranes in the cell.
THE FINE STRUCTURE OF THE DNP COMPONENT OF THE NUCLEUS
Authors: Hay Elizabeth D. , Revel J. P.
Published: May 2004

In the present investigation, the sites of deoxyribonucleic acid (DNA) synthesis and the fate of labeled deoxyribonucleoprotein (DNP) were studied in autoradiographs of ultrathin sections viewed with the electron microscope. Tritiated thymidine was employed as a label for DNA in the nuclei of proliferating cells of regenerating salamander limbs. In the autoradiographic method reported here, dilute NaOH was used to remove the gelatin of the emulsion after exposure and development. The exposed silver grains are not displaced by this treatment and the resolution of fine structure in the underlying section is greatly improved. Our observations suggest that the DNP component is a meshwork of interconnected filaments 50 to 75 A in diameter, which may be cross-linked to form what Frey-Wyssling would term a "reticular gel." The filamentous DNP meshwork is dispersed throughout the interphase nucleus during DNA synthesis, whereas in chromosomes, which are relatively inert metabolically, the meshwork is denser and is aggregated into compact masses. Dense chromatin centers in interphase nuclei are similar in fine structure to chromosomes and are also inert with respect to DNA synthesis. In the Discussion, the structure of the filamentous meshwork in chromatin is compared with that in chromosomes, and speculations are made as to the functional significance of the variations in DNP fine structure observed.
EFFECT OF TRYPSIN ON LIVER MICROSOMES
Authors: Lust J. , Drochmans P.
Published: May 2004

The effect of trypsin on the morphology of the rat liver microsomal fraction isolated by differential centrifugation has been investigated. The microsomes were incubated at 37°C and centrifuged thereafter under the conditions of their initial isolation. The trypsin-treated microsomes and the untreated controls were fixed in unbuffered osmium tetroxide and embedded first in gelose and then in methacrylate. In the trypsin-treated microsomes, there was a removal of the ribosomes from the rough vesicles. Parallel chemical determinations showed that the total nitrogen and total phosphorus of the pellet were lowered. Particles, densely stained with phosphotungstic acid (PTA) and homogeneous in appearance, were found within microsome smooth vesicles in a fluffy layer which collects on the top of the microsome pellet. The morphology of these PTA-stained particles remained unchanged after incubation with trypsin.
THE ANATOMY OF SECRETION IN THE FOLLICULAR CELLS OF THE THYROID GLAND
Authors: Wissig Steven L.
Published: May 2004

This paper reports a study by phase contrast and electron microscopy of changes observed in the thyroid gland of the rat at 1, 2, 12, and 24 hours following an injection of thyrotrophic hormone. Examination by phase contrast microscopy reveals that follicular cells contain numerous colloid droplets 1 and 2 hours after injection. By 12 and 24 hours, the colloid droplets are no longer present, and individual follicles appear to be subdividing into smaller units. The droplets are assumed to contain newly synthesized colloid, and their development was studied by electron microscopy. During the period of active secretion, increase in number of the Golgi vesicles leads to enlargement of this organelle. At its periphery small colloid droplets appear to form from large Golgi vesicles. As they form, their content becomes more adielectronic, and fine dense particles less than 75 A in diameter appear in their matrix. Small- and medium-sized droplets lying in the apical region of the cell contain numerous dense particles scattered in their moderately adielectronic content. Large, mature droplets in the same region have a relatively dielectronic content resembling follicular colloid and no longer contain dense particles. The follicular cells appear to utilize apical pseudopodia to release the content of mature droplets into the follicular lumen. Other droplet-like inclusions occur in follicular cells, but they do not seem to be directly concerned with secretion.
PLATELET DISINTEGRATION DURING CLOTTING
Authors: Rodman N. F. , Painter J. C. , McDevitt N. B.
Published: May 2004

Sequential alterations in the ultrastructure of blood platelets observed during the clotting of human platelet-rich plasma are described, with emphasis on disintegration of the platelet. As the clotting reaction proceeds, aliquots of citrated and recalcified citrated plasma are fixed by adding buffered OsO4. After recalcification a lag period of about 10 minutes is followed by an interval of rapidly occurring changes which include reorientation of cytoplasmic contents, progressive central degeneration, and disruption of the platelet limiting membrane. Shortly thereafter vesicular platelet remains are seen at the peripheries of loosely arranged and widely spaced masses of granular material. As clot retraction proceeds, these masses gradually come closer together, become more and more compact, and finally disappear. At the same time the vesicles undergo progressive disintegration until at the end of the experiment, 2 hours after recalcification, only a few are found randomly distributed in a dense clot. The significance of progressive disintegration and the origin of the vesicles observed in the later stages of the experiment are discussed in relation to clot formation and clot retraction.
ON THE STRUCTURAL CONTINUITIES OF THE TRANSVERSE TUBULAR SYSTEM OF RABBIT AND HUMAN MYOCARDIAL CELLS
Authors: Nelson Douglas A. , Benson Ellis S.
Published: May 2004

An electron microscopic study of rabbit and human myocardium provides further evidence of the existence of two distinct components of the sarcoplasmic reticulum. A thin-walled tubular system (termed longitudinal system) is arranged in anastomosing channels sur-surrounding each sarcomere and has transverse and possibly also longitudinal connections with the tubules of adjacent sarcomeres. A thick-walled tubular system traverses the myofiber transversely at the level of the Z lines of the myofibrils. The structure of these tubules very closely resembles that of deep sarcolemmal invaginations. Indeed, the membranes of the tubules appear to be continuous with the sarcolemma in favorable sections so that there seems to be an extension of the cell membrane and extracellular fluid to all depths of the myocardial fiber. Certain physiologic data which support this concept are discussed. The calculations of A. V. Hill comparing the kinetics of diffusion and the time-distance relationships between excitation and activation in frog sartorius muscle are reconsidered for cardiac muscle.