bluuebunny/crossref_metadata_2025 huggingface dataset:
LYSOSOMES IN LYMPHOID TISSUE
Authors: Bowers William E. , de Duve Christian
Published: May 2004

The density-distribution patterns of various enzymes and of labeled materials have been determined by isopycnic centrifugation in a sucrose—0.2 M KCl gradient on homogenates of lymphoid tissues from rats injected with Triton WR-1339, 14C-labeled dextran, 51Cr-labeled erythrocytes, and cortisol. The results confirm and extend the conclusion, derived from previous investigations on normal animals, that the lysosomes of lymphoid tissues form two and possibly three, distinct populations. The evidence indicates that the L19 population belongs to macrophages and the L15 group to lymphocytes. The L30 population appears to be associated with a special type of phagocyte with a high capacity for dextran storage. All three populations seem to contribute to the activities found in soluble form in homogenates of normal lymphoid tissues.
DIFFERENTIATION AND FUNCTIONAL EXPRESSION OF POTENTIAL ANTIBODY-PRODUCING CELLS IN THE PRESENCE OF CHLORAMPHENICOL
Authors: Schoenberg Melvin D. , Moore Richard D. , Weisberger Austin S.
Published: May 2004

Rabbits were immunized with diphtheria toxoid combined with complete Freund's adjuvant. Half of the animals were started on intramuscular injections of chloramphenicol 24 hr before the injection of the antigens. There was a general depression of protein synthesis in the immune system in the presence of chloramphenicol, but a greater effect on the synthesis of antibody than on the synthesis of proteins necessary for reproduction and maturation. In contrast to the finding of antibody in cells of the spleen and in the circulation of the control animals, those animals receiving chloramphenicol did not have measurable circulating antibody, and their spleens contained only a few cells with intracytoplasmic antibody late in the course of the experiment. Cytologically there was maturation of potential antibody-producing cells in the red pulp and nonfollicular white pulp of the spleen while the animals were receiving chloramphenicol. These cells developed more slowly, and were fewer and smaller than those of the control animals. They had numerous small, electron-opaque particles in their cytoplasm early in development. Ribosomes were synthesized, though fewer in number. The endoplasmic reticulum formed more slowly.
AN ELECTRON-TRANSPORT SYSTEM ASSOCIATED WITH THE OUTER MEMBRANE OF LIVER MITOCHONDRIA
Authors: Sottocasa Gian Luigi , Kuylenstierna Bo , Ernster Lars , Bergstrand Anders
Published: May 2004

Preparations of rat-liver mitochondria catalyze the oxidation of exogenous NADH by added cytochrome c or ferricyanide by a reaction that is insensitive to the respiratory chain inhibitors, antimycin A, amytal, and rotenone, and is not coupled to phosphorylation. Experiments with tritiated NADH are described which demonstrate that this "external" pathway of NADH oxidation resembles stereochemically the NADH-cytochrome c reductase system of liver microsomes, and differs from the respiratory chain-linked NADH dehydrogenase. Enzyme distributation data are presented which substantiate the conclusion that microsomal contamination cannot account for the rotenone-insensitive NADH-cytochrome c reductase activity observed with the mitochondria. A procedure is developed, based on swelling and shrinking of the mitochondria followed by sonication and density gradient centrifugation, which permits the separation of two particulate subfractions, one containing the bulk of the respiratory chain components, and the other the bulk of the rotenone-insensitive NADH-cytochrome c reductase system. Morphological evidence supports the conclusion that the former subfraction consists of mitochondria devoid of outer membrane, and that the latter represents derivatives of the outer membrane. The data indicate that the electron-transport system associated with the mitochondrial outer membrane involves catalytic components similar to, or identical with, the microsomal NADH-cytochrome b5 reductase and cytochrome b5.
MISE EN EVIDENCE ET ETUDE CYTOCHIMIQUE D'UNE PROTEINE BASIQUE EXTRANUCLEAIRE DANS LES SPERMATOZOIDES DES CRUSTACES DECAPODES
Authors: Chevaillier Philippe
Published: May 2004

Extranuclear basic proteins have been detected in the capsule of the spermatozoa of three species of decapod crustaceans (Nephrops norvegicus L., Macrura; Eupagurus bernhardus L., Anomura; Carcinus maenas Penn., Brachyura). Their properties have been studied by cytochemical methods. Their position inside the capsule of the spermatozoon has been specified with the aid of the electron microscope. Present in a constant fashion in the three species cited, their relative importance is very variable. In contrast to the refringent cone of the spermatozoon of Ascaris, which contains an acid protein, ascaradine, the capsule of the spermatozoon of the three decapod crustaceans studied contains basic proteins which we propose to designate by the general term "decapodine".
DEVELOPMENT OF FINE STRUCTURAL DAMAGE TO ALVEOLAR AND CAPILLARY LINING CELLS IN OXYGEN-POISONED RAT LUNGS
Authors: Kistler Gonzague S. , Caldwell Peter R. B. , Weibel Ewald R.
Published: May 2004

Rats were exposed to 98.5% oxygen at 765 torr for 6–72 hr. The pulmonary changes were investigated by electron microscopy and by morphometric methods. A progressive thickening of the air-blood barrier, from the normal 1.5 to 3 µ after 3 days, was due primarily to enlargement of the interstitial space by accumulation of edema which was replaced secondarily by cells and fibrin. This was accompanied by destruction of about 50% of the capillaries. Morphometric data allowed an estimate of the degree of impairment of lung function. The primary cellular damage was located in endothelial cells which underwent cytoplasmic changes and, finally, fragmentation. In contrast, the damage to the epithelial lining of alveoli was relatively scarce compared to the extensive endothelial changes. This pertained even to severely damaged lungs with 65% of the alveoli obliterated by a heterogeneous exudate. Possible causes for this apparently different reaction of epithelium (the first target cell) and endothelium to toxic oxygen effects are discussed.
CIRCADIAN PHASE RELATIONSHIPS OF THYMIDINE-3H UPTAKE, LABELED NUCLEI, GRAIN COUNTS, AND CELL DIVISION RATE IN RAT CORNEAL EPITHELIUM
Authors: Scheving Lawrence Einar , Pauly John Edward
Published: May 2004

Significant differences in the uptake of thymidine-3H, percentage of labeled cells, numbers of grains per labeled nucleus, and mitotic rate were noted in rat corneal epithelium along a 24-hr time scale. These were demonstrated by injecting subgroups of five animals every hour during a 24-hr period with thymidine-3H, sacrificing them 2 hr later, and analyzing the corneal epithelium by scintillation counter and radioautographic techniques. The increase in uptake during specific periods of the 24-hr time scale is attributed to an acceleration in the rate of DNA synthesis by individual cells and to an increase in the percentage of cells in the population actively synthesizing DNA.
THE USE OF FORMALDEHYDE-TREATED 131I-ALBUMIN IN THE STUDY OF DIGESTIVE VACUOLES AND SOME PROPERTIES OF THESE PARTICLES FROM MOUSE LIVER
Authors: Mego John L. , Bertini Francisco , McQueen J. Donald
Published: May 2004

The trichloroacetic acid-soluble radioactivity released during incubation of mouse liver particles containing intravenously injected formaldehyde-treated 131I-albumin consisted almost entirely of 131I-iodotyrosine. The material was shown to be excreted into the medium and was not due to disruption of the particles by acid. Triton X-100 or the absence of sucrose in the medium inhibited hydrolysis of the particle-associated labeled protein. This inhibition was due to disruption of the digestive vacuoles and dilution of the protein and cathepsins in the suspending medium. These results and other experimental evidence strongly suggest that the 131I-albumin-containing liver particles are digestive vacuoles. The results also establish that 131I-albumin may be used to study these vacuoles. High concentrations of sucrose (1 M) inhibited degradation of intraparticulate protein. However, 1 M salts inhibited only the rate of the digestion. Sucrose had an inhibitory effect on a crude cathepsin preparation, and salts stimulated the activity when 131I-albumin was used as substrate. The effect of high sucrose concentrations as an inhibitor of protein hydrolysis within digestive vacuoles was, therefore, most likely due principally to an inhibition of cathepsin activity within the vacuoles. The effect of salt was probably caused by a stimulation of both intra- and extra-particulate cathepsin activities, although 0.5–1.0 M KCl appeared to protect the particles.
A PECULIAR CONFIGURATION OF AGRANULAR RETICULUM (CANALICULATE LAMELLAR BODY) IN THE RAT PINEALOCYTE
Authors: Lin Huai-San
Published: May 2004

A cytoplasmic structure exhibiting a peculiar configuration of agranular reticulum has been found in the rat pinealocyte, and has been designated a canaliculate lamellar body. It consists of a number of fenestrated, flat cisternae which are closely spaced. They bear some distant resemblance to the annulate lamellae previously reported in a variety of cell types. Profiles of stacks of lamellae in a plane of section always display two distinct aspects, the surface and the cross-sectional views of flat cisternae. A surface view shows the hexagonal arrangement of pores or fenestrations. The pores in successive lamellae are aligned precisely, one behind the other, so that clear, cylindrical channels are seen running perpendicular to the lamellae as indicated by transverse sections of the lamellar stacks. Large canaliculate lamellar bodies are composed of many extended series of lamellar stacks which pursue a tortuous course and cross one another. Occasionally the canaliculate lamellar body is located deep in a nuclear invagination, which reminds one of the so-called nuclear pellets (Kernkugeln) reported by light microscopy. The functional significance of the body is unknown.
THE RENEWAL OF PHOTORECEPTOR CELL OUTER SEGMENTS
Authors: Young Richard W.
Published: May 2004

The utilization of methionine-3H by retinal photoreceptor cells has been studied by radioautographic technique in the rat, mouse, and frog. In all three species, the labeled amino acid is concentrated initially in the inner segment of the cell. Within 24 hr, the radioactive material is displaced to the base of the outer segment, where it accumulates as a distinct reaction band. The reaction band then gradually moves along the outer segment and ultimately disappears at the apex of the cell, which is in contact with the retinal pigment epithelium. These findings are interpreted to indicate that the photoreceptor cell outer segment is continually renewed, by the repeated lamellar apposition of material (membranous discs) at the base of the outer segment, in conjunction with a balanced removal of material at its apex. The outer segment renewal rate is accelerated in frogs when ambient temperature is raised, and is elevated in both frogs and rats when the intensity of retinal illumination is increased.
ABSOLUTE SENSITIVITY OF ELECTRON MICROSCOPE RADIOAUTOGRAPHY
Authors: Bachmann L. , Salpeter M. M.
Published: May 2004

A calibration method is described for measuring absolute radioautographic sensitivities under various experimental conditions. Sensitivities to 3H and 35S radiation, i.e. ratio of developed grains to radioactive decays in the specimen, were determined with Ilford L4 and Kodak NTE emulsions. The highest values obtained in monolayers of emulsion were ⅛ for 3H and 1/21 for 35S. The influence of various experimental parameters on sensitivity is described, and the possibilities for quantitative electron microscope radioautography are discussed.
CONSTANCY OF DNA CONTENT IN ADRENAL MEDULLA NUCLEI OF COLD-TREATED RATS
Authors: Cohn Norman S. , van Duijn P.
Published: May 2004

Reports of changes in DNA content of certain types of cells following exposure to conditions of stress has led to the suggestion that two kinds of DNA may be present. One is genetic DNA, and the other is called "metabolic" DNA. In a further attempt to investigate the possibility of this phenomenon, determinations of DNA content were made on Feulgen-stained nuclei of adrenal glands and kidneys in cold-treated rats. Feulgen-stained nuclei were measured by two-wavelength microspectrophotometry. Particular attention was given to the handling of the smears in hydrolysis and staining. Mean values of Feulgen-DNA contents in a total of 720 nuclei demonstrated (a) a constancy of DNA content within 2% in individual nuclei both in adrenal medulla and kidney cortex, (b) no more than an average of 2% difference in DNA content between control and experimental nuclei, and (c) no more than an average of 1.5% difference in DNA content between normal kidney cortex nuclei and normal adrenal medulla nuclei. These results confirm the view that the more precise the measurement, the more accurately the constancy rule is obeyed. Moreover, there is no support for the concept of a metabolic DNA in the rat adrenal medulla.
THE ULTRASTRUCTURE OF PHALACROCLEPTES VERRUCIFORMIS, AN UNCILIATED CILIATE PARASITIZING THE POLYCHAETE SCHIZOBRANCHIA INSIGNIS
Authors: Lom Jiři , Kozloff Eugene N.
Published: May 2004

The organization of Phalacrocleptes verruciformis is, in general, less complex than that of other ciliates, and no kinetosomes have been observed. However, there are numerous suctorial tentacles at the surface of the body, and the pellicle is characterized by close-set villus-like projections. The tentacles are very small (about 430 mµ in length, and about the same in diameter), but show the essential features of tentacles of suctorians such as Tokophrya, Podophrya, and Ephelota. Each tentacle is reinforced by eight pairs of fibrils arranged concentrically just within its wall, and contains a single missile-like body (MLB). The tentacles become attached to the cilia of the host, and serve for feeding upon the plasmatic contents of the cilia as well as for maintaining contact with the host. The MLB's originate in the endoplasm, and then migrate toward the surface and become incorporated into the tentacles. When feeding is initiated, the membrane covering the outermost nozzle-like portion of the MLB becomes continuous with the membrane of the cilium, and there are other changes in the structure of the MLB which suggest enzymatic activity. Although it appears that Phalacrocleptes is a suctorian, the complete absence of kinetosomes sets this organism apart from other members of the group.
THE EFFECTS OF MAGNESIUM ON NUCLEOSIDE PHOSPHATASE ACTIVITY IN FROG SKIN
Authors: Dahl Rolf H. , Pratley James N.
Published: May 2004

Histochemical tests, employing the Wachstein-Meisel medium, indicate that nucleoside triphosphatase activity is found predominantly in two areas of the frog skin epidermis: (1) in mitochondria, where activity is enhanced by dinitrophenol, Mg2+ dependent, but inhibited by fixation; and (2) apparently associated with cell membranes of the middle and outer portions of the epidermis, where activity is inhibited by Mg2+, unaffected by dinitrophenol, and only slightly reduced by fixation. Spectrophotometric analysis shows that Mg2+ in the medium does not increase spontaneous hydrolysis of ATP, thus obviating the possible explanation that changes in substrate concentrations in the medium lead to alterations in the "staining" distributions. It is postulated that perhaps the two enzymes differ in their requirements for substrate—one requiring the polyphosphate to be in complexed form with Mg2+, the other uncomplexed. Concentrations of Mg2+ required to inhibit cell membrane nucleoside triphosphatase activity also inhibit the electrical potential difference and short-circuit current of the frog skin. Although these observations might be taken as presumptive evidence of the cell membrane enzyme as a component of the ion pump system, because of certain dissimilarities with respect to the biochemists' "transport ATPase" and for other reasons discussed in the paper, any definite conclusions in this regard are premature.
RADIOAUTOGRAPHIC LOCALIZATION OF THE INCREASED SYNTHESIS OF PHOSPHATIDYLINOSITOL IN RESPONSE TO PANCREOZYMIN OR ACETYLCHOLINE IN GUINEA PIG PANCREAS SLICES
Authors: Hokin Lowell E. , Huebner Dorothy
Published: May 2004

A technique is described for measuring the incorporation of myo-inositol-2-3H into the lipid of various regions of the guinea pig pancreatic acinar cell by radioautography. Stimulation of enzyme secretion with either pancreozymin or acetylcholine was associated with increased graining in both the basophilic cytoplasm and the nonbasophilic cytoplasm. Kinetic studies suggested that the incorporation of myo-inositol-2-3H was stimulated independently in the two regions. Most of the increment in graining due to stimulation with pancreozymin or acetylcholine plus eserine was abolished if the tissue was extracted with 2:1 chloroform-methanol before radioautography. On chromatography of lipid extracts of pancreas, the only lipid showing a detectable increment in radioactivity on stimulation with pancreozymin was phosphatidylinositol. Thus, essentially all of the increment in graining is likely to be due to increased incorporation of tritium into phosphatidylinositol. These studies, coupled with earlier studies employing differential centrifugation, indicate that on stimulation of enzyme secretion there is increased synthesis of phosphatidylinositol in the rough-surfaced endoplasmic reticulum and in the smooth-surfaced Golgi membranes. The significance of these observations is discussed in connection with membrane circulation presumed to occur in the pancreatic acinar cell on stimulation of protein secretion. It is suggested that the increased synthesis of phosphatidylinositol may be concerned with the formation of new endoplasmic reticulum and possibly Golgi membrane to replace that which is presumably converted to membrane of the zymogen granules during intracellular protein transport.
THERMAL DEPOLARIZATION OF FLUORESCENCE FROM POLYTENE CHROMOSOMES STAINED WITH ACRIDINE ORANGE
Authors: MacInnes James W. , Uretz Robert B.
Published: May 2004

The degree of polarization of fluorescence from stretched Chironomus thummi polytene chromosomes, stained with low concentrations of acridine orange (AO), decreases with increasing temperature. The "half temperature" of this decrease (T½R) is lower than the expected DNA thermal denaturation temperature (Tm) by about 20°C. T½R is lowered as histone is removed from chromosomes. Balbiani ring regions of the fourth chromosome have T½R's much lower than other regions, and nearly as low as chromosomes which had been extensively pretreated with trypsin to remove histone and other proteins. Measurements of the thermal change in the rotational diffusion rate of AO in solution with DNA indicate that the temperature at which the DNA-AO bonding changes from a "rigid" to a "loose" mode varies with the GC percentage of the DNA, and in the same fashion as Tm, although 20°C lower.
INTRAMITOCHONDRIAL FILAMENTOUS BODIES IN THE THICK LIMB OF HENLE OF THE RAT KIDNEY
Authors: Suzuki Teruo , Mostofi F. K.
Published: May 2004

"Intramitochondrial filamentous bodies" (IMFB) were occasionally found within the matrix of some mitochondria of the thick limb of Henle of the rat kidney, but not elsewhere in the tubular system. Three types were recognized: type I, an accumulation of filaments 55 A thick; type II, a bundle of parallel filaments having the same thickness as those of type I and regular spacing, 87 A apart, from center to center; and type III, consisting of type II with regular light bands of 280 A periodicity and a helical border of prismatic tubular cristae. In addition to these, electron-opaque masses showing variable and faint substructures were found in the matrix of mitochondria. It is suggested that all these IMFB may originate from mitochondrial cristae and that type II IMFB may be an intermediate developmental form between type I and type III. After uranyl acetate staining, IMFB and the membranes of prismatic tubular cristae showed highly increased electron opacity. The literature has been reviewed for reports of intramitochondrial filamentous inclusions in various types of cells. These inclusions have been classified according to their structural characteristics and the localization in the mitochondria and compared with IMFB reported herein.
AN ELECTRON MICROSCOPIC STUDY OF NUCLEAR ELIMINATION FROM THE LATE ERYTHROBLAST
Authors: Skutelsky Ehud , Danon David
Published: May 2004

The process of expulsion of the nucleus during the transformation of the late erythroblast to reticulocyte is described. Erythroid clones taken from the spleen of lethally irradiated mice transplanted with syngeneic bone marrow were used. 10–12-day old isolated clones were fixed in glutaraldehyde, then in osmium tetroxide. Ultra-thin sections were stained with uranyl acetate and/or lead citrate before examination. Late (orthochromatic) erythroblasts develop pseudopod-like cytoplasmic protrusions into one of which the nucleus gradually penetrates, being deformed by the extrusion through the relatively narrow passage. During the whole process, mitochondria and vesicular and membranous elements are concentrated in the cytoplasm. Once outside the cell, the nucleus reassumes its rounded form. It is surrounded by a narrow rim of cytoplasm and structurally altered plasma membrane and is connected to the rest of the cell by a bridge. Elongated vacuoles appear within this bridge, with a resulting release of the enveloped nucleus which is soon phagocytized by macrophages; this leaves behind the newly formed reticulocyte. During this process, the cytoplasmic protrusions, the agglomeration of mitochondria, and the mode of separation of the nucleus from the rest of the cell are similar to those occurring in mitotic division.
THE BIOGENESIS OF MITOCHONDRIA IN SACCHAROMYCES CEREVISIAE
Authors: Clark-Walker G. D. , Linnane Anthony W.
Published: May 2004

The effects of chloramphenicol on S. cerevisiae and on a cytoplasmic respiratory-deficient mutant derived from the same strain are compared. In the normal yeast, high concentrations of chloramphenicol in the growth medium completely inhibit the formation of cytochromes a, a3, b, and c1 and partially inhibit succinate dehydrogenase formation, whereas they do not affect cytochrome c synthesis. This has been correlated with the marked reduction of mitochondrial cristae formation in the presence of the drug. In glucose-repressed normal yeast, chloramphenicol has little effect on the formation of outer mitochondrial membrane, or on the synthesis of malate dehydrogenase and fumarase. However, both these enzymes, as well as the number of mitochondrial profiles, are markedly decreased when glucose de-repressed yeast is grown in the presence of chloramphenicol. The antibiotic did not appear to affect the cytoplasmic respiratory-deficient mutant. The results have been interpreted to indicate that chloramphenicol inhibits the protein-synthesizing system characteristic of the mitochondria. Since the drug does not prevent the formation of cytochrome c, of several readily solubilized mitochondrial enzymes, or of outer mitochondrial membrane, it is suggested that these are synthesized by nonmitochondrial systems.
CORTICAL CHANGES IN GROWING OOCYTES AND IN FERTILIZED OR PRICKED EGGS OF RANA PIPIENS
Authors: Kemp Norman E. , Istock Nancy L.
Published: May 2004

The folded cortex of the growing oocyte of the frog extends as microvilli into the substance of the developing vitelline membrane and, internal to the folds, possesses a layer of cortical granules. Free ribosomes, smooth-walled vesicles, coated vesicles, tubules, and electron-opaque granules are abundant in the peripheral zone of the cortex. Mitochondria, lipochondria, pigment granules, and electron-opaque granules are conspicuous between cortical granules and in the underlying endoplasm. Yolk platelets are restricted to the endoplasm. Cortical granules contain neutral and acid mucopolysaccharides, and possibly protein. In the mature oocyte, microvilli are withdrawn and the surface folds eliminated. Cortical granules now lie close to the plasma membrane, sometimes contacting it. Fertilization or pricking causes a wave of breakdown of cortical granules lasting 1–1½ min. Breakdown begins immediately after pricking but not until about 10–15 min after insemination, because the fertilizing sperm takes that long to penetrate the jelly and vitelline membrane. Cortical granules erupt through the surface and discharge their contents into the perivitelline space. Cortical craters left at sites of eruption soon disappear, and pseudopodial protrusions retract. By 30 min after insemination, the surface of the egg is relatively smooth.
INTESTINAL TRIGLYCERIDE ABSORPTION IN THE RAT
Authors: Cardell Robert R. , Badenhausen Susan , Porter Keith R.
Published: May 2004

This report provides information on the morphology of fat absorption in rat intestinal epithelial cells. Three types of experiments were performed: (a) intubation of corn oil into fasted rats, (b) injection of physiological fatty-chyme prepared from fat-fed donor rats into ligated segments of jejunum of fasted animals, and (c) administration of electron-opaque particles in corn oil and markers given concurrently with the fat. These results support the hypothesis that fat is absorbed by selective diffusion of monoglycerides and fatty acids from micelles rather than by pinocytosis of unhydrolized triglycerides. Evidence is presented that the pits between the microvilli, previously believed to function in the transport of fat, are not involved in this process. Instead they appear to contribute their contents to lysosomes in the apical cytoplasm. Arguments are offered that the monoglycerides and fatty acids diffuse from the micelle while the latter is associated with the microvillous membrane of the absorptive cell. These micellar components penetrate the plasma membrane and diffuse into the cytoplasmic matrix where they encounter the SER. Triglyceride synthesis occurs in the SER and results in the deposition of fat droplets within its lumina. The synthesis of triglycerides and their sequestration into the SER establishes an inward diffusion gradient of monoglycerides and fatty acids.
THE REMOVAL BY PHOSPHOLIPASE C OF A LAYER OF LANTHANUM-STAINING MATERIAL EXTERNAL TO THE CELL MEMBRANE IN EMBRYONIC CHICK CELLS
Authors: Lesseps Roland J.
Published: May 2004

Fixation of embryonic chick cells (heart, neural retina, and limb bud) in the presence of lanthanum ions shows the presence of an electron-opaque layer, about 50 A thick, external to the cell membrane. This layer, designated LSM (for lanthanum-staining material), is not removable by trypsin, pronase, EDTA, DNase, α-amylase, neuraminidase, or N-acetyl-L-cysteine. However, phospholipase C, in concentrations as low as 0.001 mg/ml, succeeds in stripping the LSM from the cell surface. Heating the enzyme preparation does not inhibit this activity, but removal of divalent cations does; both of these results are consistent with the known properties of phospholipase C. The LSM is present at the cell surface in the control tissues and on cells dissociated from the tissues by proteolytic enzymes and EDTA. These results are interpreted to mean that the LSM is probably an integral part of the cell and not an extraneous coat. How this phenomenon bears on the problem of cellular adhesion is discussed, as is the possible chemical composition of the LSM.
FINE STRUCTURAL LOCALIZATION OF A BLOOD-BRAIN BARRIER TO EXOGENOUS PEROXIDASE
Authors: Reese T. S. , Karnovsky Morris J.
Published: May 2004

Horseradish peroxidase was administered to mice by intravenous injection, and its distribution in cerebral cortex studied with a recently available technique for localizing peroxidase with the electron microscope. Brains were fixed by either immersion or vascular perfusion 10–60 min after administration of various doses of peroxidase. Exogenous peroxidase was localized in the lumina of blood vessels and in some micropinocytotic vesicles within endothelial cells; none was found beyond the vascular endothelium. Micropinocytotic vesicles were few in number and did not appear to transport peroxidase while tight junctions between endothelial cells were probably responsible for preventing its intercellular passage. Our findings therefore localize, at a fine structural level, a "barrier" to the passage of peroxidase at the endothelium of vessels in the cerebral cortex. The significance of these findings is discussed, particularly with reference to a recent study in which similar techniques were applied to capillaries in heart and skeletal muscle.
FINE STRUCTURE OF THE ASEXUAL STAGES OF PLASMODIUM ELONGATUM
Authors: Aikawa Masamichi , Huff Clay G. , Sprinz Helmuth
Published: May 2004

Plasmodium elongatum, an avian malarial parasite, differs from other such parasites by infecting both the circulating red blood cells and the hematopoietic cells. The exoerythrocytic development of P. elongatum occurs mainly in these red cell precursors. The fine structure of the asexual stages of P. elongatum has been studied in the bone marrow and peripheral blood of canaries and compared with that of the asexual stages of other avian malarial parasites. With minor differences, the merozoites of P. elongatum possess the same organelles as those in the exoerythrocytic merozoites of P. fallax and the erythrocytic stages of P. cathemerium, P. lophurae, P. fallax, and P. gallinaceum. The developmental sequence is also essentially similar to that of other avian malarial parasites, in that upon entry into a new host cell, the dedifferentiation, growth, and redifferentiation phases take place. However, we have found some important differences in the feeding mechanism of P. elongatum. The cytostome is involved in the ingestion of host cell cytoplasm in both exoerythrocytic and erythrocytic stages, in contrast to P. fallax, in which the cytostome is inactive in the exoerythrocytic stages. In P. elongatum, host cell cytoplasm is ingested through the cytostome, and "boluses" are formed and incorporated into a large digestive vacuole. Subsequently, the digestion of the boluses takes place in this digestive vacuole. Thus, in regard to the function of the cytostome, the exoerythrocytic stages of P. elongatum appear to be closely related to the erythrocytic stage which has a feeding mechanism similar to that of the erythrocytic stage of other avian malarial parasites.
STUDIES OF THE MITOCHONDRIAL ENERGY-TRANSFER SYSTEM OF BROWN ADIPOSE TISSUE
Authors: Lindberg O. , De Pierre J. , Rylander E. , Afzelius B. A.
Published: May 2004

An investigation of the mechanisms of norepinephrine action and heat production in brown adipose tissue from newborn rabbits has been carried out. Data obtained with the use of biochemical techniques has been correlated with morphological data from electron microscopy. Norepinephrine was found to stimulate the respiration of brown fat in vitro. Inhibitors of glycolysis abolish this effect, whereas inhibitors of oxidative phosphorylation do not, at least not to the same extent. Brown fat is readily permeable to added Krebs cycle intermediates. Substrate level phosphorylation, but no electron transport-coupled phosphorylation, could be demonstrated in isolated mitochondria. It is suggested that the rate of fatty acid oxidation is limited by the availability of phosphate acceptor systems which break down ATP formed at the substrate level and thus provide ADP for further substrate level phosphorylation. The theory of respiratory control by the action of reesterification of fatty acids is discussed in the light of these findings. Under the electron microscope, brown fat mitochondria are characterized by their large size, tightly packed cristae, and by the different types of granules in the matrix. No elementary particles are seen when the mitochondria are examined by the negative-staining technique. The absence of electron transport-coupled phosphorylation together with the apparent absence of elementary particles seems to be of particular significance.
STUDIES ON THE MICROTUBULES IN HELIOZOA
Authors: Tilney Lewis G. , Porter Keith R.
Published: May 2004

When specimens of Actinosphaerium nucleofilum are placed at 4°C, the axopodia retract and the birefringent core (axoneme) of each axopodium disappears. In fixed specimens, it has been shown that this structure consists of a highly patterned bundle of microtubules, each 220 A in diameter; during cold treatment these microtubules disappear and do not reform until the organisms are removed to room temperature. Within a few minutes after returning the specimens to room temperature, the axonemes reappear and the axopodia begin to reform reaching normal length 30–45 min later. In thin sections of cells fixed during the early stages of this recovery period, microtubules, organized in the pattern of the untreated specimens, are found in each reforming axopodium. Reforming axopodia without birefringent axonemes (and thus without microtubules) are never encountered. From these observations we conclude that the microtubules may be instrumental not only in the maintenance of the axopodia but also in their growth. Thus, if the microtubules are destroyed, the axopodia should retract and not reform until these tubular units are reassembled. During the cold treatment short segments of a 340-A tubule appeared; when the organisms were removed from the cold, these tubular segments disappeared. It seems probable that they are one of the disintegration products of the microtubules. A model is presented of our interpretation of how a 220-A microtubule transforms into a 340-A tubule and what this means in terms of the substructure of the untreated microtubules.
CHROMOSOME PULVERIZATION IN HUMAN BINUCLEATE CELLS FOLLOWING COLCEMID TREATMENT
Authors: Kato Hatao , Sandberg Avery A.
Published: May 2004

Under the influence of Colcemid, a substantial number of binucleate human cells from a line infected with herpes-like virus was found to possess pulverized chromosomes. Although this abnormality was also detected in untreated binucleate cells, the increase in the number of pulverized cells after the addition of Colcemid was too striking to be explained by accumulation of spontaneously occurring cells in response to the mitotic inhibition by Colcemid. Furthermore, the induction of pulverization may be dependent upon Colcemid concentration. These findings imply an involvement of Colcemid in the mechanism of pulverization induction in the system studied. When tritiated thymidine was added to the culture medium simultaneously with Colcemid, the majority of binucleate cells with an intact and a pulverized chromosome set incorporated this isotope into the pulverized set only. This obviously suggests that the nuclei in the binucleate cell are asynchronous in DNA synthesis, and that this asynchrony is intimately related to the induction of the pulverization phenomenon. It seems very probable that the late S phase in the late synthesizing nuclei represents a critical stage at which damage to the chromosomes most readily occurs.
A MORPHOLOGICAL STUDY OF HALOBACTERIUM HALOBIUM AND ITS LYSIS IN MEDIA OF LOW SALT CONCENTRATION
Authors: Stoeckenius Walther , Rowen Robert
Published: May 2004

The reported absence of a cell wall in halobacteria cannot be confirmed. Improved fixation techniques clearly show a cell wall-like structure on the surface of these cells. A stepwise reduction of the salt concentration causes the release of cell wall material before the cell membrane begins to disintegrate. The cell membrane breaks up into fragments of variable but rather small size, which are clearly different from a 4S component reported by others to be the major breakdown product of the cell membrane. It appears more likely that the 4S component arises from the dissolution of the cell wall. A residue of large membranous sheets remains even after prolonged exposure of halobacteria envelopes to distilled water. The lipids in these sheets do not differ significantly from the lipids in the lysed part of the cell membrane. The sheets, however, contain a purple-colored substance, which is not present in the lysed part. The easily sedimentable residue that remains after lysis of the cells or envelopes in distilled water also contains "intracytoplasmic membranes" with unusual structural characteristics. They can also be identified in sections through intact bacteria or envelope preparations. Their function is at present unknown but seems to be related to the formation of gas vacuoles in these organisms.
DNA-PROTEIN BINDING IN INTERPHASE CHROMOSOMES
Authors: Himes Marion
Published: May 2004

The metachromatic dye, azure B, was analyzed by microspectrophotometry when bound to DNA fibers and DNA in nuclei with condensed and dispersed chromatin. The interaction of DNA and protein was inferred from the amount of metachromasy (increased ß/α-peak) of azure B that resulted after specific removal of various protein fractions. Dye bound to DNA-histone fibers and frog liver nuclei fixed by freeze-methanol substitution shows orthochromatic, blue-green staining under specific staining conditions, while metachromasy (blue or purple color) results from staining DNA fibers without histone or tissue nuclei after protein removal. The dispersed chromatin of hepatocytes was compared to the condensed chromatin of erythrocytes to see whether there were differences in DNA-protein binding in "active" and "inactive" nuclei. Extraction of histones with 0.02 N HCl, acidified alcohol, perchloric acid, and trypsin digestion all resulted in increased dye binding. The amount of metachromasy varied, however; removal of "lysine-rich" histone (extractable with 0.02 N HCl) caused a blue color, and a purplish-red color (µ-peak absorption) resulted from prolonged trypsin digestion. In all cases, the condensed and the dispersed chromatin behaved in the same way, indicating the similarity of protein bound to DNA in condensed and dispersed chromatin. The results appear to indicate that "lysine-rich" histone is bound to adjacent anionic sites of a DNA molecule and that nonhistone protein is located between adjacent DNA molecules in both condensed and dispersed chromatin.
ULTRASTRUCTURE OF THE SPOON TYPE SYNAPTIC ENDINGS IN THE NUCLEUS VESTIBULARIS TANGENTIALIS OF THE CHICK
Authors: Hinojosa Raúl , Robertson J. David
Published: May 2004

The fine structure of the "spoon" type synaptic endings of the chick tangential nucleus was studied with the electron microscope. These endings often measure ∼18 µ in length by ∼3–4 µ in width. The axoplasm of the endings contains very few synaptic vesicles, a large number of neurofilaments oriented parallel to the long axis of the nerve fiber, and microtubules and numerous mitochondria. The synaptic membrane complex shows areas of localized occlusion of the synaptic cleft with the formation of an external compound membrane. It has not been decided whether these areas have a disc shape; their length measures between 0.04 and 0.47 µ. The five-layer pattern characteristic of an external compound membrane is shown in specimens fixed with formalin—OsO4, glutaraldehyde—acrolein—OsO4, and acrolein KMnO4 but it does not appear in the glutaraldehyde-OsO4-fixed specimens. The over-all thickness of the external compound membrane varies depending upon the fixative used. The synaptic clefts in the regions between the external compound membrane discs are widened and measure ∼300 A. A condensation of dense material occurs in pre- and postsynaptic cytoplasms all along the synaptic membrane complex. The morphological relationships described in the spoon endings are suggestive of electrical transmission.
FINE STRUCTURE OF DIVISION IN CILIATE PROTOZOA
Authors: Jenkins R. A.
Published: May 2004

The mitotic, micronuclear division of the heterotrichous genus Blepharisma has been studied by electron microscopy. Dividing ciliates were selected from clone-derived mass cultures and fixed for electron microscopy by exposure to the vapor of 2% osmium tetroxide; individual Blepharisma were encapsulated and sectioned. Distinctive features of the mitosis are the presence of an intact nuclear envelope during the entire process and the absence of centrioles at the polar ends of the micronuclear figures. Spindle microtubules (SMT) first appear in advance of chromosome alignment, become more numerous and precisely aligned by metaphase, lengthen greatly in anaphase, and persist through telophase. Distinct chromosomal and continuous SMT are present. At telophase, daughter nuclei are separated by a spindle elongation of more than 40 µ, and a new nuclear envelope is formed in close apposition to the chromatin mass of each daughter nucleus and excludes the great amount of spindle material formed during division. The original nuclear envelope which has remained structurally intact then becomes discontinuous and releases the newly formed nucleus into the cytoplasm. The micronuclear envelope seems to lack the conspicuous pores that are typical of nuclear envelopes. The morphology, size, formation, and function of SMT and the nature of micronuclear division are discussed.
THE LOSS OF KINETOPLASTIC DNA IN TWO SPECIES OF TRYPANOSOMATIDAE TREATED WITH ACRIFLAVINE
Authors: Steinert M. , van Assel Suzanne
Published: May 2004

The effects of acriflavine on two species of Trypanosomatidae, Crithidia luciliae and Trypanosoma mega, have been investigated. It has been observed that kinetoplastic (i.e. mitochondrial) DNA is lost in a high percentage of acriflavine-treated cells. Resting flagellates, from stationary-phase or hemin-deficient cultures, are considerably more resistant to the acridine than are flagellates from a log-phase culture. When the kinetoplast has retained some DNA and still remains visible in stained smears, it appears reduced in size, and its ultrastructure is extremely abnormal: the DNA fibrils, clearly visible in normal kinetoplasts, are condensed; they appear as an electron-opaque, apparently homogeneous mass, separated from the membranes by a space of low electron-opacity. Analyses of DNA extracts, with high speed centrifugation in CsCl density gradients, revealed that the satellite band, presumably kinetoplastic DNA, is lost by trypanosomes grown for 5 days in the presence of acriflavine. Radioautography was used to study the effects of acriflavine on thymidine-3H incorporation in C. luciliae. At the concentration which affects the kinetoplast specifically, the dye produces an 87% inhibition of thymidine incorporation in this organelle. The kinetics of this inhibition suggest a direct effect on replication. No decrease in incorporation occurs in the nucleus. These results lead to the conclusion that loss of kinetoplastic DNA is due to continued growth and cell division in the absence of kinetoplastic DNA replication. Several hypotheses are discussed concerning the specificity of the dye's action upon the replication of extrachromosomal DNA.
OCCURRENCE, ISOLATION, AND CHARACTERIZATION OF POLYRIBOSOMES IN YEAST
Authors: Marcus Leon , Ris H. , Halvorson H. O. , Bretthauer R. K. , Bock R. M.
Published: May 2004

This report details the procedural requirements for preparing cell-free extracts of yeast rich in polyribosomes. This enabled us to demonstrate the occurrence of polyribosomes in yeast, to show their role in protein synthesis, and to devise methods for their resolution and isolation. When certain precautions are met (the use of log phase cells, rapidly halting cell growth, gentle methods of disruption, sedimentation through exponential density gradients, etc.), individual polyribosome size classes ranging up to the heptosome can be fractionated and separated from their nearest neighbors. Larger size classes are resolved partially among themselves, free of smaller polyribosomes. This was confirmed by extensive electron micrographic studies of material from the various fractions obtained upon density gradient centrifugation of yeast extracts. Modifications of the gradients and procedure should allow fractionation and isolation of the larger polyribosomes, including those containing polycistronic messages. Yeast polyribosomes are disaggregated to single ribosomes by longer term grinding, cell disruption by the French pressure cell, the Hughes press, or by incubation with dilute RNAse. Yeast polyribosomes are active in the incorporation of amino acids into polypeptide; the single ribosomes exhibit only slight activity. The latter activity is probably due to the presence of a small fraction of monosomes still containing mRNA. Poly-U stimulates amino acid incorporation only in the single ribosomes.
THE MECHANISM OF ACTION OF COLCHICINE
Authors: Borisy G. G. , Taylor E. W.
Published: May 2004

The majority of the colchicine-3H bound by tissue culture cells (KB or Hela) was found to be present as a noncovalent complex with a macromolecule which appears in the soluble fraction after homogenization. Similar binding was demonstrated in vitro and was confined to a component of the soluble fraction. The binding-equilibrium constant and the kinetic constants were essentially the same in vivo and in vitro. Bound radioactivity was reisolated and shown to be present in a molecule with the same chromatographic behavior and specific antimitotic activity as colchicine. In vitro assay of binding activity of a variety of cells and tissues showed a correlation with the presence of microtubules. High binding activity was given by dividing cells, mitotic apparatus, cilia, sperm tails, and brain tissue. Binding to extracts of slime mold or to purified muscle proteins was very low or undetectable. The binding site had a sedimentation constant of 6S and it is suggested that the protein is a subunit of microtubules.
INTRACELLULAR TRANSPORT OF SECRETORY PROTEINS IN THE PANCREATIC EXOCRINE CELL
Authors: Jamieson James D. , Palade George E.
Published: May 2004

In the previous paper we described an in vitro system of guinea pig pancreatic slices whose secretory proteins can be pulse-labeled with radioactive amino acids. From kinetic experiments performed on smooth and rough microsomes isolated by gradient centrifugation from such slices, we obtained direct evidence that secretory proteins are transported from the cisternae of the rough endoplasmic reticulum to condensing vacuoles of the Golgi complex via small vesicles located in the periphery of the complex. Since condensing vacuoles ultimately become zymogen granules, it was of interest to study this phase of the secretory cycle in pulse-labeled slices. To this intent, a zymogen granule fraction was isolated by differential centrifugation from slices at the end of a 3-min pulse with leucine-14C and after varying times of incubation in chase medium. At the end of the pulse, few radioactive proteins were found in this fraction; after +17 min in chaser, its proteins were half maximally labeled; they became maximally labeled between +37 and +57 min. Parallel electron microscopic radioautography of intact cells in slices pulse labeled with leucine-3H showed, however, that zymogen granules become labeled, at the earliest, +57 min post-pulse. We assumed that the discrepancy between the two sets of results was due to the presence of rapidly labeled condensing vacuoles in the zymogen granule fraction. To test this assumption, electron microscopic radioautography was performed on sections of zymogen granule pellets isolated from slices pulse labeled with leucine-3H and subsequently incubated in chaser. The results showed that the early labeling of the zymogen granule fractions was, indeed, due to the presence of highly labeled condensing vacuoles among the components of these fractions.